CM 1.2 contents · Chinese Medicine (CM) Journal Information SUBSCRIPTIONS The Chinese Medicine (Online at Scientific Research Publishing, ) is published quarterly by Scientific Research

Chinese Medicine, 2010, 1, 31-57 Published Online September 2010 in SciRes (http://www.SciRP.org/journal/cm/)

Copyright © 2010 SciRes. CM

TABLE OF CONTENTS

Volume 1 Number 2 September 2010

Antimicrobial Activity of Polyalthia longifolia (Sonn.) Thw. var. Pendula Leaf Extracts Against 91

Clinically Important Pathogenic Microbial Strains

S. Chanda, R. Nair………………………………………………………………………………………………………………31

Research on Depression, Anxiety and Memory of Subhealth after Practicing Jianshenqigong-Wuqinxi

Y. Wang, C. L. Chen, Z. Zhang……………………………………………………………………………………………………39

Briefly Discuss on the Philosophize of Meridian Research

L. H. Wei, S. H. Xiao……………………………………………………………………………………………………………43

The Wu Xing Theory and Homeostatic Interaction of Organs

Y. V. Albegov, D. V. Butenko, L. N. Butenko………………………………………………………………………………45

The Effect of EDTA Chelation Therapy in Symptomatic Coronary Heart Disease:

An Observational Study

C. D. A. Goonasekera, R. Tennakoon, P. N Rajakrishna, G. A. Gunasena, C. R. Wanniarachchi,

A. B. Yatawatta, U. A. D. D. Munidasa…………………………………………………………………………………………49

Prevalence of Glucose-6-Phosphate Dhydrogenase Deficient Neonates and Cost Effective of

Enzyme Screening in Tehran, Iran

H. Dahifar, A. Ghorbani, M. Ghods………………………………………………………………………………………………55

Chinese Medicine (CM)

Journal Information

SUBSCRIPTIONS

The Chinese Medicine (Online at Scientific Research Publishing, www.SciRP.org) is published quarterly by Scientific Research

Publishing, Inc., USA.

Subscription rates: Print: $50 per issue.

To subscribe, please contact Journals Subscriptions Department, E-mail: [email protected]

SERVICES

Advertisements

Advertisement Sales Department, E-mail: [email protected]

Reprints (minimum quantity 100 copies)

Reprints Co-ordinator, Scientific Research Publishing, Inc., USA.

E-mail: [email protected]

COPYRIGHT

Copyright©2010 Scientific Research Publishing, Inc.

All Rights Reserved. No part of this publication may be reproduced, stored in a retrieval system, or transmitted, in any form or by

any means, electronic, mechanical, photocopying, recording, scanning or otherwise, except as described below, without the

permission in writing of the Publisher.

Copying of articles is not permitted except for personal and internal use, to the extent permitted by national copyright law, or under

the terms of a license issued by the national Reproduction Rights Organization.

Requests for permission for other kinds of copying, such as copying for general distribution, for advertising or promotional purposes,

for creating new collective works or for resale, and other enquiries should be addressed to the Publisher.

Statements and opinions expressed in the articles and communications are those of the individual contributors and not the statements

and opinion of Scientific Research Publishing, Inc. We assumes no responsibility or liability for any damage or injury to persons or

property arising out of the use of any materials, instructions, methods or ideas contained herein. We expressly disclaim any implied

warranties of merchantability or fitness for a particular purpose. If expert assistance is required, the services of a competent

professional person should be sought.

PRODUCTION INFORMATION

For manuscripts that have been accepted for publication, please contact:

E-mail: [email protected]

Chinese Medicine, 2010, 1, 31-38 doi:10.4236/cm.2010.12006 Published Online September 2010 (http://www.SciRP.org/journal/cm)


Antimicrobial Activity of Polyalthia longifolia (Sonn.) Thw. var. Pendula Leaf Extracts Against 91 Clinically Important

Pathogenic Microbial Strains

Sumitra Chanda*, Rathish Nair Phytochemical, Pharmacological and Microbiological Laboratory

Department of Biosciences, Saurashtra University, Rajkot, Gujarat, India E-mail: [email protected]

Received May 21, 2010; revised July 21, 2010; accepted July 26, 2010

Abstract The methanol, acetone and 1,4-dioxan fractions of leaves of Polyalthia longifolia (Sonn.) Thw. were evalu-ated for antibacterial and antifungal activity. 91 clinically important strains were used for the study which were both clinical isolates as well as identified strains. Piperacillin and gentamicin were used as standards for antibacterial assay, while nystatin and flucanazole were used as standards for antifungal assay. The antibac-terial activity was more pronounced against gram positive bacterial and fungal strains. Poor activity was shown against gram negative bacterial strains studied. Keywords: Antibacterial, Antifungal, Polyalthia longifolia, Clinical Isolates, Organic Solvent Extracts

1. Introduction Due to the increasing development of drug resistance in human pathogens as well as the appearance of undesir-able effect on certain antimicrobial agents, there is a need to search for new agents. The world health organi-zation in 1997 suggested that effective locally available plants be used as substitutes for drugs. Research work on medicinal plants be intensified and information on these plants be exchanged. This thought will go a long way in the scientific exploration of medicinal plants for the benefit of man and is likely to decrease the dependence on importance of drugs [1]. Polyalthia longifolia (An-nonaceae) is a tree, which is widely distributed in Bang-ladesh, Srilanka and throughout the hotter parts of India [2]. In India, the seeds of this plant were used as febri-fuge [3]. Literature survey revealed that most of the plants of annonaceae family contain antitumor and anticancer principles [4,5]. The bark is also used as a febrifuge in the Balasore district of Orissa [6]. The ex-tract of stem bark and the alkaloids isolated from this were found to demonstrate a good antibacterial and antifungal activities [7]. In the present study, antim-icrobial potentiality of the P. longifolia leaves was in-vestigated against a few clinically isolated as well as standard microbial cultures.

2. Materials and Methods

2.1. Plant Material Polyalthia longifolia (Sonn.) Thw. (Annonaceae) leaves were collected in May, 2004 from Rajkot in the State of Gujarat (Western India) and identified by comparison with specimens (PSN 4) available at the Herbarium of the Department of Biosciences, Saurashtra University, Rajkot, Gujarat, India.

2.2. Extraction Leaves of P. longifolia were collected, air dried and then powdered in a homogenizer and 10 gm was used for dif-ferent solvent extractions (Methanol, Acetone, N, N-dimethylformamide); the sample was extracted in sol-vent kept on a rotary shaker overnight, and then the fil-trate was collected and centrifuged at 5000 rpm. The solvent was then evaporated to dryness under reduced pressure and the extracted compound left was used for the antimicrobial assay. The percentage yield of 1, 4-dioxan, methanol and acetone extracts were 20.56, 29.30 and 13.52 respectively.

Microorganisms Studied 91 clinically important mi-crobial strains which included 23 gram positive, 56 gram

S. CHANDA ET AL.


32

negative and 15 fungal strains were studied for the an-timicrobial activity. These strains included both clinical isolates as well as identified strains. The details of the microorganisms used are shown in Table 1. Table 1. List of bacterial and fungal strains studied for an-timicrobial assay.

Sr. Strain Specimen

Gram Positive bacteria

1 Staphylococcus spps. [10] Sputum

2 Staphylococcus aureus [11] Pus

3 Staphylococcus aureus [13] Urine


5 Staphylococcus spps. [26] Pus

6 Staphylococcus aureus [34] Sputum

7 Staphylococcus aureus [35] Tracheal

8 Staphylococcus aureus [36] Tracheal

9 Staphylococcus spps. [44] Sputum

10 Staphylococcus aureus [47] Ear swab

11 Staphylococcus aureus [48] Sputum



14 Staphylococcus aureus ATCC 25923 -

15 Staphylococcus epidermidis ATCC 12228 -

16 Staphylococcus subfava NCIM 2178 -

17 Bacillus cereus ATCC 11778 -

18 Bacillus subtilis ATCC 6633 -

19 Bacillus megaterium ATCC 9885 -

20 Micrococcus flavus ATCC 10240 -

Gram negative bacteria

21 Pseudomonas spps. [15] Sputum

22 Pseudomonas spps. [17] Pus

23 Pseudomonas fluorescence [18] Pus

24 Pseudomonas spps. [25] Urine


26 Pseudomonas aeruginosa [30] Sputum

27 Pseudomonas spps. [37] Tracheal

28 Pseudomonas aeruginosa [38] Pus

29 Pseudomonas spps. [39] Wound swab

30 Pseudomonas fluorescence [40] Tracheal





35 Pseudomonas spps. [50] Tracheal secretion

36 Pseudomonas fluorescence [59] Urine

37 Pseudomonas aeruginosa ATCC 27853 -

38 Pseudomonas testosteroni NCIM 5098 -

39Pseudomonas pseudoalcaligenes ATCC 17440

-

40 E.coli [14] Pus

41 E.coli [16] Urine

42 E.coli [21 ] Urine



45 E.coli [28] Pus


47 E.coli [32 ] Stool

48 E.coli [33] Pus


50 E.coli [45] Pus

51 E. coli [51] Urine

52 E. coli [58] Vaginal swab

53 E. coli [60] Urine

54 E. coli [61] Blood

55 E. coli ATCC 25922 -

56 Enterobacter spps. [1] Tracheal

57 Enterobacter spps. [8] Tracheal

58 Enterobacter aerogenes ATCC 13048 -

59 Klebsiella spps [6] Urine

60 Klebsiella spps [19] Sputum

61 Klebsiella aerogenes [52] Pus

62 Klebsiella spps. [54] Urine

63 Klebsiella aerogenes [57] Urine

64 Klebsiella pneumoniae NCIM 2719 -

65 Proteus mirabilis [4] Wound swab

66 Proteus spps. [53] Pus

67 Proteus mirabilis NCIM 2241 -

68 Proteus vulgaris NCTC 8313 -

69 Proteus morganii NCIM 2040 -

70 Providencia rettgeri [5] Pus

71 Citrobacter spps. [20] Pus

S. CHANDA ET AL.


33

72 Citrobacter freundii [29] Pus

73 Citrobacter freundii ATCC 10787 -

74 Alcaligenes fecalis ATCC 8750 -

75 Salmonella typhimurium ATCC 23564 -

Fungus

76 Candida albicans [1] Urine

77 Candida albicans [2] Sputum

78 Candida spps. [3] Sputum

79 Candida spps. [4] Sputum

80 Candida spps. [5] Urine

81 Candida albicans ATCC 2091 -

82 Candida albicans ATCC 18804 -

83 Candida glabrata NCIM 3448 -

84 Candida tropicalis ATCC 4563 -

85 Candida apicola NCIM 3367 -

86 Cryptococcus neoformans ATCC 34664 -

87 Cryptococcus luteolus ATCC 32044 -

88 Trichosporan beigelii NCIM 3404 -

89 Aspergillus flavus NCIM 538 -

90 Aspergillus candidus NCIM 883 -

91 Aspergillus niger ATCC 6275 -

2.3. Preparation of Samples

Methanol, acetone and 1,4-dioxan extracts were dis-solved in 100% DMSO at a concentration of 25 mg/ml and 12.5 mg/ml and were used as working stocks respec-tively. Sterile discs (Hi-media Labs) were impregnated with 20 µl of the stock solution. Gentamicin (10 µg/disc) and Piperacillin (100 µg/disc) for bacteria; nystatin (100 units/disc) and flucanazole (10 µg/disc) (Himedia Labs) for fungus were used as positive control and pure DMSO was used as a negative control.

2.4. Antimicrobial Study Antimicrobial activity was performed by agar disc diffu-sion method [8,9]. The bacterial strains were grown in nutrient broth while fungal strains were grown in MGYP (Malt glucose yeast peptone) broth. Mueller Hinton agar no. 2 was the media used to study the antibacterial sus-ceptibility while Sabroaud agar was used to study the antifungal susceptibility test. The cultures were grown for 24 hours, and the turbidity of the culture was main-tained according to the 0.5 MacFarland standards. The inoculum’s size was 1 × 108 cells/ml.

2.5. Agar Disc Diffusion

The media (Mueller Hinton Agar No.2 and MRS media) and the test bacterial cultures were poured into Petri dishes (Hi-Media). The test strain (200 µl) was inocu-lated into the media (inoculums size 108 cells/ml) when the temperature reached 40-42°C. The test compound (20 µl) was impregnated in to sterile discs (7 mm) (Hi-Media) and was then allowed to dry. The disc was then intro-duced into medium with the bacteria. The plates were incubated overnight at 37°C for bacterial strains and 28°C for fungal strains. The experiment was performed under strictly aseptic conditions. Microbial growth was determined by measuring the diameter of the zone of inhibition. The experiment was performed in triplicates and the mean values of the result are shown in Table 2.

3. Results and Discussion Herbal medicine in developing countries is commonly used for the traditional treatment of health problems [10]. In recent years multiple drug resistance in human patho-genic microorganisms has developed due to the indis-criminate use of commercial antimicrobial drugs com-monly used in the treatment of infectious diseases, mak-ing it a global growing problem [11-13]. In addition to this problem antibiotics are sometimes associated with adverse effects on host including hypersensitivity, im-mune suppression and allergic reactions [14]. Therefore there is a need to develop alternative antimicrobial drugs for the treatment of infections obtained from various sources such as medicinal plants [15,16]. In the present study, P. longifolia leaf extracts extracted in 1, 4-dioxan (PDE), methanol (PME) and acetone extracts (PAE) were investigated at two different concentrations for their antimicrobial potentiality against 91 clinically important microbial strains. All the three extracts (PDE, PME and PAE at 500 µg/disc concentration) were active against 95% of the total gram positive bacterial strains studied. PDE was active against 18.18% of the total gram nega-tive bacterial strains studied (active against 21% of Pseudomonas spps., 33.3% of Enterobacter spps., 16% of Klebsiella spps., 33.3% of Proteus spps. and 66.6% of Citrobacter spps.). PME and PAE were active against 12.72% of the total gram negative bacterial strains stud-ied. P. aeruginosa is most common pathogen of im-muno-compromised individuals [17]. Infections caused by Pseudomonas spps. are among the most difficult to treat with conventional antibiotics. Both PME and PAE were active against 5.26% of the Pseudomonas spps. and 66.6% of Enterobacter spps. PME was active against 33.3% of Klebsiella spps. and Proteus spps., while PAE was active against 66.6% of Klebsiella spps. and Proteus spps. studied. Salmonellosis is an important public

S. CHANDA ET AL.


34

Table 2. Antimicrobial activity of Polyalthia longifolia against 91 clinically important microbial strains (inhibition zone in mm).

Control Extracts Antibiotics Sr. No.

Strain DMSO PDE-500 PME-500 PAE-500 PDE-250

PME-250

PAE-250

G Pc Fu Ns

Gram Positive bacteria

1 Staphylococcus spps. [10]

- 15 ± 0.58

12.67± 0.33

10 ± 0.58

10 ± 0.58

11 ± 0.58

14.67±0.88

- - NT NT

2 Staphylococcus aureus [11]

- 13 ± 0.58

12 ± 0.58

10 ± 0.58

11 ± 0.58

12 ± 1.15

12 ± 0.58

18.67± 0.33

17.33 ±

0.33 NT NT


- 12 ± 1.15

12 ± 2.31

9 ± 0.58

- - - - - NT NT


- 11 ± 0.58

12 ± 1.73

9 ± 0.58

12 ± 1.73

9 ± 1.15

- - - NT NT

5 Staphylococcus spps [26]

- 16.5 ± 0.28

11± 0.58

13± 0.58

15± 0.58

13 ± 1.73

14 ± 1.73

- - NT NT


- 15.5 ± 0.28

9 ± 0.58

13 ± 0.58

14 ± 0.58

9 ± 0.58

10 ± 1.15

- - NT NT


- 22± 0.28

12 ± 0.28

14 ± 0.58

17± 0.58

8 ± 0.58

11 ± 0.58

- - NT NT


- 13 ± 0.58

9.67 ± 0.33

13 ± 0.58

12.67 ± 0.88

- 8.67±0.88

- - NT NT

9 Staphylococcus spps [44]

- 13 ± 0.58

10.33 ± 0.33

12.33 ± 0.33

18 0.58

11 ± 0.58

10.67 ±

0.66

14.67± 0.33

- NT NT


- 12 ± 3.21

10.67 ± 2.03

11 ± 2.31

9 ± 1.15

8.67 ±0.88

12 ± 2.89

- - NT NT


- - - - - - -

20.67± 0.33

- NT NT


- 13.67 ± 0.33

12.67 ± 0.33

13.67 ± 0.33

11.67 ± 0.33

- - - - NT NT


- 15.67 ±

0.33 10 ± 1.53

11.67 ± 0.88

12.33 ± 0.33

10.33 ±

1.76

12.67 ±

0.33

10.33± 0.33

- NT NT

14 Staphylococcus aureus ATCC 25923

- 13 ± 0.58

8 ± 0.58

9 ± 0.58

14.33 ± 0.88

9.5 ±0.28

9 ± 0.58

- - NT NT

15 Staphylococcus epidermidis ATCC 12228

- 14.5 ± 2.60

16 ± 2.69

13 ± 0.5813.5 ± 0.87

13 ± 0.57

12 ± 1.73

- - NT NT

16 Staphylococcus subfava NCIM 2178

- 10.5 ± 0.29

11.5 ± 1.44

12.5 ± 0.28

13 ± 2.319.5 ± 0.28

9.5 ± 0.28

- 20.17

± 0.44

NT NT

17 Bacillus cereus ATCC 11778

- 29.5 ± 0.28

21.5 ± 0.28

25. ± 0.58

25 ± 2.3121 ± 0.58

25 ± 0.58

20.17 ± 0.16

18.83 ±

0.16 NT NT

18 Bacillus subtilis ATCC 6633

- 26.5 ± 1.44

21.5 ± 1.44

23.5 ± 0.28

25 ± 0.5821 ± 0.58

21 ± 0.58

18.33 ± 0.33

17.83 ±

0.93 NT NT

19 Bacillus megaterium ATCC 9885

- 14 ± 0.58

10.5 ± 0.28

12.5 ± 0.28

13 ± 0.5811 ± 0.58

10.5 ± 0.28

- - NT NT

20 Micrococcus flavus ATCC 10240

- 12.5 ± 0.28

10.5 ± 0.28

11 ± 2.3111.5 ± 0.28

9 ± 0.58

9 ± 0.58

27.67 ± 0.33

12.67 ±

0.33 NT NT

Gram negative bacteria

NT NT

21 Pseudomonas spps. [15]

- - - - - - -

14 ± 0.58

- NT NT


- 8 ± 0.58

- - - - 12 ± 2.89

- - NT NT

23 Pseudomonas fluorescence [18]

- 8± 0.58

- - - - 12± 2.89

- - NT NT


- - - - - - - - - NT NT

S. CHANDA ET AL.


35


- - - - - - - - - NT NT

26 Pseudomonas aeruginosa [30]

- -- - - - - - 16.67±

0.67 - NT NT


- -- - - - - - - -- NT NT

28 Pseudomonas aeruginosa [38]

- - - - - - - 19.67±

0.33 - NT NT


- - - - - - - - - NT NT

30 Pseudomonas fluorescence [40]

- - - - - - - - - NT NT


- - - - - - - - - NT NT


- - - - - - - - - NT NT


- - - - - - - - - NT NT


- 8 ± 0.58

- - - - 8 ± 0.58

20 ± 0.58

- NT NT


- - - - - - - - - NT NT

36 Pseuodmonas fluorescence [59]

- - - - - - - - - NT NT

37 Pseudomonas aeruginosa ATCC 27853

- - - - - - - 17 ± 1.15

12.33 ±

0.66 NT NT

38 Pseudomonas testosteroni NCIM 5098

- - - - - - - 22.33 ± 0.66

- NT NT

39 Pseudomonas pseudoalcaligenes ATCC 17440

- 8.5 ±

0.86 14 ± 1.73

10.5 ± 0.86

- - - 19.33 ± 0.6

- NT NT

40 E.coli [14] - - - - - - - - - NT NT 41 E.coli [16] - - - - - - - - - NT NT 42 E.coli [21 ] - - - - - - - - - NT NT 43 E.coli [22] - - - - - - - - - NT NT 44 E.coli [24] - - - - - - - - - NT NT

45 E.coli [28] - 17± 0.33

NT NT

46 E.coli [31] - - - - - - - - - NT NT

47 E.coli [32 ] - - - - - - - 21± 0.58

- NT NT

48 E.coli [33] - - - - - - - - - NT NT

49 E.coli [41] - - - - - - - 18.67±

0.33 - NT NT

50 E.coli [45] - - - - - - - - - NT NT

51 E. coli [51] - - - - - - - 20.33±

0.33 - NT NT

52 E. coli [58] - - - - - - - - - NT NT 53 E. coli [60] - - - - - - - - - NT NT 54 E. coli [61] - - - - - - - - - NT NT

55 E. coli ATCC 25922

- - - - - - - 17.83

± 0.16

14.5 ±

0.50 NT NT

56 Enterobacter spps. [1]

- - - - - - - - - NT NT

57 Enterobacter spps. [8]

- 15 ± 0.58

12 ± 0.58

14.33 ± 1.20

13 ± 0.58

12.33 ±

0.88

12 ± 1.15

19.67± 0.88

- NT NT

58 Enterobacter aerogenes ATCC 13048

- - 8.5 ± 0.86

15 ± 0.58

- - - - - NT NT

59 Klebsiella spps [6] - - - - - - - 22± 0.58

- NT NT

60 Klebsiella spps [19]

- - - - - - - - - NT NT

61 Klebsiella aero-genes [52]

- - - 8 ± 0.58

13 ± 1.73

11 ± 2.08

- - - NT NT

S. CHANDA ET AL.


36

62 Klebsiella spps. [54]

- - - - - - - - - NT NT

63 Klebsiella aero-genes [57]

- - - - - - - - - NT NT

64 Klebsiella pneu-moniae NCIM 2719

- 12 ± 0.58

12 ± 0.58

10.5 ± 0.28

10.5 ± 0.86

10.5 ±0.86

11 ± 0.58

- 24.67

± 0.33

NT NT

65 Proteus mirabilis [4]

- - - - - - - - 14± 0.58

NT NT

66 Proteus spps. [53] - - - - - -- - - - NT NT

67 Proteus mirabilis NCIM 2241

- 10.5 ± 0.86

10.5 ± 0.28

9.5 ± 0.86

- - - 18.67 ± 0.33

- NT NT

68 Proteus vulgaris NCTC 8313

- - 9 ± 1.15 - - - - 18 ± 1.00

- NT NT

69 Proteus morganii NCIM 2040

- 9 ± 0.58 - - 8 ± 0.58 - - - - NT NT

70 Providencia rett-geri [5]

- - - - - - - - - NT NT

71 Citrobactor spps [20]

- 8 ± 0.58

8 ± 0.58

8 ± 0.58

- - - - - NT NT

72 Citrobactor freundii [29]

- - - - - - - 12.33±

0.33 - NT NT

73 Citrobactor freundii ATCC 10787

- 11 ± 0.58 - - 11.5 ± 0.28

10 ± 0.58

9.5 ± 0.28

- - NT NT

74 Alcaligenes fecalis ATCC 8750

- - - - - - - 18.33 ± 0.66

- NT NT

75 Salmonella ty-phimurium ATCC 23564

- - - - - - - 18.5 ± 0.28

- NT NT

Fungus

76 Candida albicans [1]

- 7.5 ± 0.29

8 ± 0.58

- 7.5 ± 0.29

10.5 ±0.29

10 ± 0.58

NT NT - 11.33

± 0.33

77 Candida albicans [2]

- - - 10 ± 0.58

13.33 ± 0.88

9 ± 0.58

- NT NT - 18 ±0.58

78 Candida spps. [3] - - - 9.5 ± 0.29

14.33 ± 0.66

12.5 ±0.86

8 ± 0.58

NT NT - 14 ±0.58

79 Candida spps. [4] - 11 ± 2.13

10.5 ± 2.02

11.5 ± 2.06

8 ± 0.58

8.5 ±0.29

12.5 ±0.86

NT NT - 14 ±0.58

80 Candida spps. [5] - 7.5 ± 0.29

8.5 ± 0.29

9.5 ± 0.29

7.5 ± 0.29

- - NT NT - 10 ±0.58

81 Candida albicans ATCC 2091

- 11.5 ± 2.60

11 ± 2.31

8 ± 0.58

7.5 ± 0.29

7.5 ±0.29

10.5 ±2.02

NT NT 17.67

± 0.33

13 ±0.58

82 Candida albicans ATCC 18804

- 10.5 ± 0.29

8 ± 0.58

- - 11 ± 0.58

15 ± 1.15

NT NT - 14.33

± 0.33

83 Candida glabrata NCIM 3448

- - - - - - - NT NT 39.67

± 0.88

22 ±0.58

84 Candida tropicalis ATCC 4563

- - - 7.5 ± 0.29

11 ± 0.58

12 ± 0.58

9.5 ± 0.29

NT NT - 8.33

± 0.33

85 Candida apicola NCIM 3367

- 23 ± 3.60

26 ± 0.58

28 ± 1.15

25.33 ± 0.88

24 ± 0.58

21.66 ±

0.33 NT NT -

21.33 ±

0.88

86 Cryptococcus neoformans ATCC 34664

- 7.5 ± 0.29

8 ± 0.58

- - - 9.5 ± 1.4

NT NT 21.33

± 0.33

17 ±0.58

87 Cryptococcus luteolus ATCC 32044

- 14 ± 0.58

11.5 ± 0.86

11 ± 1.15

9.5 ± 1.44

8.5 ±0.86

8.5 ± 0.88

NT NT 23.66

± 0.88

17.66 ±

0.88

88 Trichosporan beigelii NCIM 3404

- 12 ± 0.58

13 ± 1.73

10.5 ± 2.02

- - - NT NT - -

89 Aspergillus flavus NCIM 538

- - - - 14.67 ±

4.34 22 ± 0.58

10.33 ± 2.02

NT NT - -

S. CHANDA ET AL.


37

90 Aspergillus can-didus NCIM 883

- 10.5 ± 0.29

9 ± 1.15

11 ± 0.58

- - - NT NT - -

91 Aspergillus niger ATCC 6275

- - - - 11 ± 2.31

- - NT NT - -

Mean ± SEM, n = 3, zone includes disc diameter 7 mm; G – Gentamicin (10 µg/disc), Pc – Piperacillin (100 µg/disc), Ns – Nystatin (100 units/disc), Fu – Fluconazole (10 µg/disc); PME – Methanol extract, PAE – Acetone extract, PDE – Dioxan extract, DMSO – Dimethylsulphoxide.

health problem worldwide. Salmonella infection is pri-marily associated with gastroenteritis. This illness poses a more serious health risk to sensitive populations in the community such as the elderly, young and the immuno-compromised, where hospitalization may be required. All the three extracts were inactive against E. coli, A. fecalis and S. typhimurium. Several antimycotic drugs are available at present, its use is limited by a number of factors such as low potency, poor solubility, emergence of resistance strains and drug toxicity. Therefore there is distinct need for the discovery of new, safer and more effective antifungal agents. Candida species have be-come a common cause of hospital acquired infections and a large number of patients die as a result of invasive Candidal infections [18]. All the three extracts were ac-tive against 62.5% of the total fungal strains studied. The three extracts were active against A. candidus while it was inactive against the remaining two moulds (A. flavus and A. niger) studied. The details of the results are given elaborately in Table 2. From the results obtained, it seems that the antibacterial action of the extracts is more pronounced on gram positive than on gram negative bacteria and these findings correlate with the observa-tions of previous screenings of medicinal plants for an-timicrobial activity, where most of the active plants showed activity against gram positive strains only [19- 21]. This difference in susceptibility is because of the difference in cell wall structure of gram positive and gram negative organisms. The lipopolysaccharide con-tent of gram negative bacteria makes them resistant to plant extracts while the peptidoglycan layer of gram positive bacteria is not an effective permeability barrier. 4. Conclusions All the extracts of P. longifolia exhibited the highest rates of antimicrobial activity against gram positive and fungal strains studied. Therefore, it is concluded that P. longifolia extracts should further be studied phyto-chemically to elucidate the active principle in the leaf, which can be used as a leading antibacterial (specific for gram positive) and antifungal agent. 5. Acknowledgements

Financial support from Department of Special assistance (DSA) project, New Delhi and supply of clinical isolates by Micro Care and Spandan Diagnostic Laboratories,

Rajkot are gratefully acknowledged. 6. References [1] C. K. Amadou, “Promoting Alternative Medicine,” Africa

Health Journal, 1998, Vol. 2, pp. 20-25.

[2] J. D. Hooker and C. B. Clarke, “Flora of British India, Vol. 1,” L. Reeve and Co. Ltd., London, 1875, pp. 1-741.

[3] K. Raghunathan and M. K. Mitra, “Pharmacognosy of Indigenous Drugs, Vol. 1,” Central Council for Research in Ayurveda and Siddha, New Delhi, 1985, pp. 127-139.

[4] S. K. Chakrabarti and B. Mukherjee, “Search for Anti-cancer Drug from Indian Medicinal Plants,” Indian Journal of Medical Research, Vol. 56, No. 4, 1968, pp. 445-455.

[5] K. Yamaguchi, H. Kinora, S. Natori, Ito, K. Nissbimoio, K. Bando, D. Mizuno and M. Ishignoo, “Screening Tests for Antitumor Activity of Asian Medicinal Herbs I,” Ya-kugaku Zashi, 1964, Vol. 84, pp. 373-377.

[6] K. R. Kirtikar and B. D. Basu, “Indian Medicinal Plants,” In: Annonaceae, 2nd Edition, Lalit Mohan Basu, Leader Road, Allahabad, India, Vol. 1, pp. 1993, pp. 72-73.

[7] C. M. Hasan, S. N. Islam and M. Ahsan, “Antibacterial Activity of Stem Bark of Polyalthia longifolia,” Dhaka University Studies, Part E, 1988, Vol. 4, pp. 63-66.

[8] A. W. Bauer, W. M. M. Kirby, J. C. Sherries and M. Truck, “Antibiotic Susceptibility Testing By Standard Single Disc Diffusion Method,” American Journal of Clinical Pathology, Vol. 45, No. 4, 1966, pp. 426-493.

[9] J. Parekh and S. Chanda, “Antibacterial and Phyto-chemical Studies on Twelve Species of Indian Medicinal Plants,” African Journal of Biomedical Research, Vol. 10, No. 2, 2007, pp. 175-181.

[10] M. J. Martinez, J. Betancourt, N. Alanso-Gonzalea and A. Jauregui, “Screening of Some Cuban Medicinal Plants for Antimicrobial Activity,” Journal of Ethnopharmacology, Vol. 52, No. 3, 1996, pp. 171-174.

[11] J. E. Loper, M. D. Henkels, R. G. Roberts, G. G. Grove, M. J. Willett and T. J. Smith, “Evaluation of Streptomy-cin, Oxytetracycline and Copper Resistance of Erwinia amylavora isolated from pear orchards in Washington State,” Plant Disease, Vol. 75, No. 3, 1991, pp. 287-290.

[12] J. Davis, “Inactivation of Antibiotics and Dissemination of Resistance Genes,” Science, Vol. 264, No. 5157, 1994, pp. 375-382.

[13] R. F. Service, “Antibiotics That Resist Resistance,” Sci-ence, Vol. 270, No. 5237, 1995, pp. 724-727.

[14] I. Ahmad, Z. Mehmood and F. Mohammad, “Screening of Some Indian Medicinal Plants for their Antimicrobial Properties,” Journal of Ethnopharmacology, Vol. 62, No.

S. CHANDA ET AL.


38

2, 1998, pp. 183-193.

[15] A. M. Clark, “Natural Products as Resource of New Drugs,” Pharmaceutical Research, Vol. 13, No. 8, 1996, pp. 1133-1141.

[16] G. A. Cordell, “Biodiversity and Drug Discovery a Sym-biotic Relationship,” Phytochemistry, Vol. 55, No. 66, 2000, pp. 463-480.

[17] J. R. Zgoda and J. R. Porter, “A Convenient Microdilu-tion Method for Screening Natural Products against Bac-teria and Fungi,” Pharmaceutical Research, Vol. 39, No. 3, 2001, pp. 211-225.

[18] T. J. Walsh, J. W. Hathorn, J. D. Sobel, W. G. Merz, V. Sanchez, S. N. Maret, H. R. Buckley, M. A. Pfaller, R. Schaufele, C. Sliva, E. Navarro, J. Lecciones, P. Chandrasekar, J. Lee and P. A. Pizzo, “Detection of Cir-culating Candida enolase by Immunoassay in Patients with Cancer and Invasive Candidiasis,” New England

Journal of Medicine, Vol. 324, No. 15, 1991, pp. 1026- 1031.

[19] R. M. Herrera, M. Perez, D. A. Martin-Herrera, R. Lopez-Garcia and R. M. Rabanal, “Antimicrobial Activ-ity of Extracts from Plants Endemic to the Canary Is-lands,” Phytotherapy Research, Vol. 10, No. 6, 1996, pp. 364-366.

[20] N. A. A. Ali, W. D. Julich, C. Kusnick and U. Lindequist, “Screening of Yemeni Medicinal Plants for Antibacterial and Cytotoxic Activities,” Journal of Ethnopharmaco- logy, Vol. 74, No. 2, 2001, pp. 173-179.

[21] R. Nair and S. Chanda, “In vitro Antimicrobial Activity of Psidium guajava L. Leaf Extracts against Clinically Important Pathogenic Microbial Strains,” Brazilian Jour- nal of Microbiology, Vol. 38, No. 3, 2007, pp. 452-458.



Research on Depression, Anxiety and Memory of Subhealth after Practicing Jianshenqigong-Wuqinxi

Yan Wang1, Changle Chen 2, Zhang Zhang3 1School of Medical Instrument and Food Engineering, University of Shanghai for Science and Technology,

Shanghai, China 2Shanghai Qigong Institute, Shanghai, China

3Dongzhimen Hospital Affiliated to Beijing University of Chinese Medicine, Beijing, China E-mail: [email protected], [email protected]

Received May 26, 2010; revised July 27, 2010; accepted August 8, 2010

Abstract Objective: To study the effect of practicing Jianshenqigong-wuqinxi on subhealth problems including de-pression, anxiety, and memory loss. Methods: 1) 80 cases with subhealth problem were randomly divided into two groups. 40 cases practicing Jianshenqigong-wuqinxi were used as test group, and the other 40 un-practicing cases were used as control group. 2) The test group was practising Jianshenqigong-wuqinxi one hour in the morning five times per week for three months. 3) Scores of subhealth status such as depression and anxiety were evaluated once every two weeks, while the memory test was performed once a month. 4) Data analysis: SPSS11.0 software was used for statistical analyses. Data were shown in mean ± SEM. Dif-ference was considered significant if the t-test P value is less than 0.05. Results: 1) Scores of subhealth state and depression were significantly lower in test group than those of control group. The beneficial effects of practicing Jianshenqigong-wuqinxi were more obvious for the subjects in the test group who had practiced Jianshenqigong-wuqinxi for two to twelve weeks (P < 0.05). 2) Scores of anxiety test were significantly lower in test group than those of control group after practicing Jianshenqigong-wuqinxi for six to twelve weeks (P < 0.05). 3) Scores of memory were significantly higher in test group than those of control group (P < 0.05). Conclusions: 1) Practicing Jianshenqigong-wuqinxi could promote health in people who have sub-health mental problems. 2) Practicing Jianshenqigong-wuqinxi could relieve depression, anxiety while en-hance memory. Keywords: Subhealth, Depression, Anxiety, Jianshenqigong-Wuqinxi

1. Introduction

Subhealth is a kind of status between health and disease, which has no precise definition but is generally defined as low physical activity and adaptability regularly occur. It has been reported from WHO that there are more than 70 percent people in the world are in subhealth status [1,2]. In China the subhealthy people are about 60 to 70 percent, most of which are between 35 to 45 years old. The Chinese in subhealth conditions have exceeded 7 million according to recent data [3,4]. For those people they are often disturbed by bad feelings such as depres-sion, anxiety, panic, short breath, listlessness, dizziness, headache, insomnia, etc. Jianshenqigong-wuqinxi mim-ics five animals’ appearances and actions such as tiger, deer, bear, monkey, and crane. And it is a kind of mildly

intense aerobic exercise, which makes vasodilatation, regulates autonomic nerve, increases venous return and enhances body sensibility. It was supposed that practic-ing Jianshenqigong-wuqinxi could relieve the bad feel-ings characteristic of subhealthy people. To discover the mechanism of physiological and psychological changes of Jianshenqigong·wuqinxi on subheathy people, we ob-serve the effects of practicing Jianshenqigong-wuqinxi on depression, anxiety, and memory loss of subhealthy people. 2. Methods

1) 80 cases (not hospitalized patients) with subhealth selected from Tianlin community and Muyang hospital were randomly divided into 2 groups. Subjects in the test

Y. WANG ET AL.


40

group (40 cases) were practicing Jianshenqigong ·wuqinxi, while subjects in the control group (40 cases) did not practice Jianshenqigong·wuqinxi.

2) The test group was practicing Jianshenqigong- wuqinxi one hour in the morning five times per week for three months.

3) Scores of subhealth staus such as depression and anxiety were evaluated once every two weeks, while the memory test was evaluated once a month.

4) Index of test: a) The diagnosis of subhealth was according to the

Fukuda complete text of revised case definition in Ame- rican control disease center (CDC) in 1994 [5]. Nine items were tested in diagnosis of subhealth. The subjects should answer each item as yes or no. If the answer was yes, 1 score would be added (except the first item). Two scores would be added for the first item. After all the items were finished, the person whose score more than 6 was considered as in subhealth status.

b) Self-rating Depression Scale (SDS) and Self-Rating Anxiety (SAS) tables were used in depression and anxi-ety test accordingly. Twenty items were tested in calcu-lation of scores of SDS and SAS. Each item has 4 grades. The first grade is ‘no’, the second is ‘some time’, the third is ‘many time’ and the forth is ‘most or all the time’. SDS scores less than 40 were ‘normal’, between 40 and 60 were ‘mild depress’, more than 60 were ‘heavy de-press’. SAS scores less than 40 were ‘normal’ between 40 and 60 were ‘mild anxiety’ more than 60 were ‘heavy anxiety’.

c) Memory test: a list of random numbers or letters was presented on a computer screen at the rate of one per two seconds. The subjects being tested were asked to recall the items in order, and if they are correct, the computer would record the length of the numbers or let-ters as the memory of the subjects. The test began with two numbers, increasing until the subjects committed twice errors. Recognizable patterns (for example 2, 4, 6, 8) should be avoided [6].

5) Data analysis: SPSS11.0 software was used. Data were shown in mean ± SEM. Difference was considered significant at P < 0.05.

3. Rerults

1) Scores of subhealthy state (Table 1) were signifi-cantly lower than that of control group. The beneficial effects of practicing Jianshenqigong-wuqinxi were more obvious for the subjects in the test group who had prac-ticed Jianshenqigong-wuqinxi for two to twelve weeks (P < 0.05). There were also significant difference of sub-health scores in control group before and after two to twelve weeks (P < 0.05).

2) Scores of anxiety (Table 2) of test group were sig-

Table 1. Scores of subhealth after practicing Jianshen-qigong-wuqinxi for twelve weeks.

Time（weeks） Control group Test group

0 6.45 ± 0.36 6.40 ± 0.40

2 6.1 ± 0.7☆ 5.20 ± 1.45☆△

4 6.18 ± 0.76☆ 4.95 ± 1.74☆△

6 6.03 ± 1☆ 4.65 ± 1.52☆△

8 5.88 ± 1.19☆ 4.53 ± 1.54☆△

10 5.75 ± 1.16☆ 4.38 ± 1.24☆△

12 5.78 ± 1.48☆ 4.5 ± 1.51☆△ ☆There was significant difference when compared with before practicing (P < 0.05); △There was significant difference when compared with control group (P < 0.05).

Table 2. Anxiety scores of subhealth after practicing Jian-shenqigong-wuqinxi for twelve weeks.


0 36.8 ± 7.86 36.8 ± 7.38

2 36.5 ± 6.74 35.5 ± 7.91

4 36 ± 8.12 34.7 ± 7.14

6 35.8 ± 8.08 32.5 ± 6.77☆△

8 36.0 ± 8.12 32.1 ± 7.55☆△

10 35.5 ± 8.31 32.4 ± 7.79☆

12 35.6 ± 8.52 31.9 ± 7.52☆△ ☆There was significant difference when compared with before practicing (P < 0.05); △There was significant difference when compared with control group (P < 0.05).

nificantly lower than that of control group. The benefi-cial effects of practicing Jianshenqigong-wuqinxi were more obvious for the subjects in the test group who had practiced Jianshenqigong-wuqinxi for six to twelve weeks (P < 0.05). There was no significant difference of anxiety scores in control group before and after two to twelve weeks (P > 0.05).

3) Scores of depression (Table 3) of test group were significantly lower than that of control group. The bene-ficial effects of practicing Jianshenqigong-wuqinxi were more obvious for the subjects in the test group who had practiced Jianshenqigong-wuqinxi for two to twelve weeks (P < 0.05). There was no significant difference of anxiety scores in control group before and after two to twelve weeks (P > 0.05).

4) Scores of memory (Table 4) were higher in test group than that of control group. The beneficial effects of practicing Jianshenqigong-wuqinxi were more obvi-ous for the subjects in the test group who had practiced Jianshenqigong-wuqinxi for twelve weeks (P < 0.05). There was no significant difference of memory scores in control group before and after two to twelve weeks (P > 0.05).

Y. WANG ET AL.


41

Table 3. Depression scores of subhealth after practicing Jianshenqigong-wuqinxi for twelve weeks.


0 40.68 ± 9.12 40.40 ± 8.86

2 39.90 ± 9.88 36.08 ± 9.47☆△

4 39.85 ± 9.98 35.78 ± 9.52☆△

6 39.53 ± 8.86 35.73 ± 8.79☆△

8 39.38 ± 8.88 35.60 ± 8.90☆△

10 38.92 ± 7.92 35.06 ± 9.15☆△

12 38.64 ± 8.61 35.34 ± 8.46☆△ ☆There was significant difference when compared with before practicing (P < 0.05); △There was significant difference when com-pared with control group (P < 0.05).

Table 4. Memory (digits) test of subhealth after practicing Jianshenqigong-wuqinxi for twelve weeks.


0 7.28 ± 2.53 7.13 ± 1.94

4 7.28 ± 2.53 7.63 ± 1.92

8 7.18 ± 2.23 8.2 ± 2.61

12 7.17 ± 2.47 8.86 ± 2.00☆△ ☆There was significant difference when compared with before prac-ticing (P < 0.05); △There was significant difference when compared with control group (P < 0.05).

4. Conclusions 1) Practicing Jianshenqigong-wuqinxi could promote health in people who have subhealth mental problems.

2) Practicing Jianshenqigong·wuqinxi could relieve bad feelings such as depression, anxiety of subhealth people and enhance memory. 5. Discussions Many researchers have observed the effect of practicing Jianshenqigong-wuqinxi on different people. Jing-mei Wu found after 3-month practicing, Jianshenqigong- wuqinxi could improve the immunity of the people, and the immunity ability of woman practitioners grew even faster [7]. Ding-hai Yu found after 6-month exercise of Jianshenqigong-wuqinxi, the NK cell activity of exercis-ers in the experimental group was higher than that before exercise and control group [8]. But no scientific work studied the effect of Jianshenqigong-wuqinxi on sub-health people and no people observed the physiological and psychological changes of practicers. We discovered the effects of practicing Jianshenqigong-wuqinxi on subhealth mental problems. Subhealth status is charac-terized by panic, short breath, dizziness, headache, in-somnia, etc.

Subhealth is the major cause of many diseases [9].

People with subhealth problems could become severely ill later in their lifetime, which were highly correlated with their subhealth problems [10]. Therefore optimal intervention of subhealth problems is necessary. In our experiment we found that there were significant differ-ence of subhealth scores between Jianshenqigong- wuqinxi practitioners and controls (Table 1), which sug-gested Jianshenqigong-wuqinxi could promote mental health of people who have subhealth problems. Com-parison with controls, more cases of subhealthy practic-ers became mentally healthy after practicing Jianshen-qigong-wuqinxi for 3 months, suggesting the beneficial effects of Jianshenqigong-wuqinxi.

It was reported that tai chi could regulate psychologi-cal health of subhealthy people, and relieve bad feelings such as lassitude, memory loss, irritable, anxiety, insom-nia, etc [11]. Jianshenqigong-wuqinxi is a kind of mildly intense aerobic exercise just like tai chi. From our ex-periment we found practicing Jianshenqigong-wuqinxi could relieve bad feelings such as anxiety (Table 2) and depression (Table 3), while enhance memory (Table 4). The reasons might be as follows: a) Jianshenqigong- wuqinxi is a kind of exercise that can regulate bad feel-ings such as anxiety and depression through releasing of biological macromolecules such as adrenocorticotropic hormone, cortisol, catecholamine, opioid peptides, etc. These molecules are believed to be beneficial to psycho-logical status [12,13]. b) Jianshenqigong-wuqinxi has multiple mental stages. Psychological self-regulating was required for practicing each stage of Jianshenqigong- wuqinxi. For example, when practicing the stage of ‘ti-ger-xi’, the practitioners will mimic a powerful tiger. When practicing the stage of ‘deer-xi’, the practitioners will mimic a gentle deer. Such changes of psychological stages were reported to be beneficial to reducing bad feelings [14].

It is therefore concluded that the effects of practicing Jianshenqigong-wuqinxi have significant beneficial ef-fects on improving the overall mental health of people suffering from subhealth problems. 6. Acknowledgements

This work was supported by grants awarded to Chen Chang-le from the General Administration of Sport of China (QG200093), and to Yan Wang from Science and Technology Commission of Shanghai Municipality (Outstanding Youth Foundation of University of Shang-hai for Science and Technology). 7. References [1] L. D. Wang, “Chronic Disease and Subhealth in China:

Y. WANG ET AL.


42

The Effect and the Guideline of Prevention and Treat-ment,” Chinese Medical Journal, Vol. 83, No. 12, June 2003, pp.1031-1034.

[2] X. L. Wang, Y. H. Huo, J. Li, X. S. Zhao and R. Luo, “Evaluation of the Questionnaire for Subhealth Status Survey Based on the Symptoms in Traditional Chinese Medicine,” Journal of Southern Medical University, Vol. 27, No. 2, February 2007, pp. 160-163.

[3] T. F. Wang, J. J. Wang, X. L. Xue, P. Han, Y. J. Zhang, G. R. Li, X. Y. Wu, Y. Zhao, L. L. Tang, Y. Y. Liu, C. L. Sui, C. Fu, Y. M. Shang and B. Zhou, “Distribution Charac-teristics of Traditional Chinese Medicine Syndromes and their Elements in People with Subhealth Fatigue,” Jour-nal of Chinese Integrative Medicine, Vol. 8, No. 3, March 2010, pp. 220-223.

[4] Y. H. Huo, X. L. Wang, J. Li, C. Y. Zhu, X. M. Sun, X. S. Zhao, C. Zhang and R. Luo, “Analysis of Clinical Mani-festations of Subheath Status in a Medical College of Guangdong Province,” Journal of Southern Medical Uni-versity, Vol. 27, No. 4, April 2007, pp. 448-449.

[5] K. Fukuda, S. E. Straus, I. Hickie, M. C. Sharpe, J. G. Dobbins and A. Komaroff, “The Chronic Fatigue Syn-drome: A Comprehensive Approach to its Definition and Study. International Chronic Fatigue Syndrome Study Group,” Annals of Internal Medicine, Vol. 121, No. 12, December 1994, pp. 953-959.

[6] G. Miller, “The Magical Number Seven, Plus or Minus Two: Some Limits on Our Capacity for Processing In-formation,” Psychological Review, Vol. 101, No. 2, April 1994, pp. 343-352.

[7] J. M. Wu and D. H. Yu “Research on Effects of Practic-ing ‘Wuqinxi’ on the Outer Sub-Group of T-Cell of the Middle-Aged and Old People,” Journal of Beijing Sport

University, Vol. 29, No. 8, August 2006, pp. 1074-1076.

[8] D. H. Yu and J. M. Wu, “Effects of Exercising Build-ing-Up Qigong-Wuqinxi on Middle-Aged and Old Peo-ple’s NK Cell Activity,” Journal of Shanghai University of Sport, Vol. 32, No. 1, January 2008, pp. 56-59.

[9] H. W. Jiang, “Integrative Medical Research Thinking on the Constitution-Subhealth-Disease Relation Developing Axis,” Chinese Journal of Integrated Traditional and Western Medicine, Vol. 29, No. 9, September 2009, pp. 836-837.

[10] C. X. Fan, S. Y. Wang, L. Zhu, Y. J. Xiao and S. B. Ma, “Study on the Prevalence and Risk Factors of ‘Subhealth’ Status in College and University Staff in Guangdong Province,” Zhonghua Liu Xing Bing Xue Za Zhi, Vol. 24, No. 9, September 2003, pp. 774-777.

[11] H. Yin and J. Yang, “Rehabilitative Effects of Taichi Exercise on the Subhealthy Status of College Students,” Journal of Clinical Rehabilitative Tissue Engineering Research, Vol. 39, No. 11, 2007, pp. 7991-7992.

[12] M. A. Nies and C. L. Motyka, “Factors Contributing to Women’s Ability to Maintain a Walking Program,” Journal of Holistic Nursing, Vol. 24, No. 1, March 2006, pp. 7-14.

[13] B. B. Simen, C. H. Duman, A. A. Simen and R. S. Du-man, “TNFalpha Signaling in Depression and Anxiety: Behavioral Consequences of Individual Receptor Target-ing,” Biological Psychiatry, Vol. 59, No. 9, February 2006, pp. 775-785.

[14] A.-H. Peng and Z. Yan, “Analysis of Epidemiological Status and Countermeasures for Sub-Health Problem among Occupational Poputation in China,” Modern Pre-ventive Medicine, Vol. 36, No. 14, November 2009, pp. 2617-2619.



Briefly Discuss on the Philosophize of Meridian Research

Lianhai Wei, Shuhua Xiao Tianjin University of TCM—The College of Acupuncture and Moxibustion, Tianjin, China

E-mail: {xiaoshuhua129, weilhai001}@163.com Received June 16, 2010; revised August 9, 2010; accepted August 16, 2010

Abstract Meridian theory is an important part of the TCM system. Since 1950s, we have already studied the essence of meridian for more than a half century, but there was no substantive progress. In this article the author point that, in order to determine the concept and scope of meridian, it’s necessary to have a briefly discussion on the philosophize of meridian research, and the most important is the Meridian research in future should be multi-angle, multi-level, three-dimensional and so on. Keywords: Essence of Meridian, Philosophize, Study of Three-Dimensional

1. Introduction Meridian theory is an important part of the TCM system, which studies human physiological functions and patho-logical changes of Meridian, as well as the relationship between it and the viscera. Meridian theory has guided the diagnosis and treatment of TCM for thousand years, and it has made significant contribution to the people of the world .Meridian research, or exactly providing tradi-tional meridian theory a modern explanation, is an un-avoidable problem in the process of internationalization of TCM, while thoughts and methods of scientific re-search is the key to success, therefore it’s necessary for us to do some philosophical thinking about meridian research.. 2. Current Situation of Meridian Research Meridian research is a big engineering, which involves excavation, inheritance, development of TCM theory and the modernization of TCM, so it’s bound to face various difficulties for it to achieve substantive progress only by a small number of research institutions. Dr. Wang Guangjun [1] who works in China Academy of Chinese Medical Science pointed out that, there are two problems in meridian research, one is the educational background of researchers, at present the main researchers are non-Chinese medicine workers, the other one is that me-ridian research in different fields is confusion, for exam-ple TCM areas, Chinese and Western Integrative Medi-cine areas, Biological Sciences areas and so on, the ob-jects and purposes in different areas are also distinct. Dr.

Wang said that the meridian research in biological field had made rapid progress at abroad.

3. Object of Meridian Research

In the book “Outline of the History of Chinese Acu-puncture” [2], Professor Huang Longxiang indicated that the quintessence of meridian theory lies in revealing a fact that there are special relations between surface and internal organs of the body. The key question now is what the facts are, what “tentative explanation” to those facts in ancient Chinese meridian theory are. It is indeed problem. As we all known facts are objective. When people describe them it always mixed their subjective feelings, furthermore people of different ideological ba-sis also has different feelings to the facts, and so there are many different genres. School of Yin and Yang, Taoist School and Legalist School are representatives of different Schools.

Therefore it’s the depth, breadth, and accuracy of cog-nition to the “fact” of meridians which determined the object of meridian research. Definition of meridian con-cept and scope are very important, and it will be much appropriate for workers who expert in TCM and Chinese philosophy to accomplish this task. However, philoso-phers seldom participated in the present study of merid-ian. If we can not define the concept and scope of “fact” in meridian accurately, experts in various fields can only do meridian research freely, and then it will just like blind men touching an elephant. There won’t be much achievement, if any.

L. H. WEI ET AL.


44

4. Methods of Meridian Research To find out the role of the meridian and it’s guidance to clinical physiology and pathology is also very important, after determining the concept and scope of meridian, after all, meridian can not explain all the complex phe-nomenon of human life. Some clinical experts had done a very good job in this respect, such as, Dr. Du Yuanhao [3] who works in the affiliated hospital of Tianjin Uni-versity of Traditional Chinese Medicine, after years of clinical research and statistical analysis, has put forward four kinds spectrum of disease in terms of acupuncture. Grade I, all diseases which can be cured by acupuncture, such as peripheral facial paralysis and hysteria. In the treatment acupuncture can intervene the important link of diseases; Grade II, acupuncture can act on the main symptoms and signs of diseases, but the key link of dis-eases is difficult to eliminate completely, or the disease pathology is complex, acupuncture only act on one of the important aspects of diseases, then combination with other treatment is necessary. For example, in the patho-logic process of cerebral vascular diseases, hypertension, hyperlipidemia, high blood sugar and blood theology, dynamics and other factors are all involved. Acupuncture has a good effect in improving cerebral circulation and body functions, but the recovery of disease need inter-vention of many other treatments. Grade III, acupuncture can only relieve symptoms of disease partly, such as atrophic gastritis and other acute appendicitis. Grade IV, it includes diseases in which the effect of acupuncture is imprecise, or new treatment is highly effective and acu-puncture rarely used, the former, such as various types of cancer, tuberculosis, gonorrhea, the latter as malaria. Because the base of acupuncture treatment is meridian theory, so the research of Dr. Du should be vested in the clinical study.

After determining “body and use” of the meridian, it seems appropriate for us to start from the “use”, that is, the physiological and pathological phenomena which can be explained by meridian theory, particularly disease suitable for acupuncture treatment, to explore the “body” of meridian, or exactly “substance of meridian”. The guiding concept of meridian research is historical mate-rialism and dialectical materialism, and we must reso-lutely abandon mechanical materialism, let alone mysti-cal and idealist. The attitude of meridian research should be open, and we also should keep pace with the times. TCM belonging to the life sciences has never refused the intervention of other sciences, from ancient astronomy to modern computer information technology have been in- filtrated into TCM already.

5. Prospect of Meridian Research Meridian research in future should be multi-angle, multi- level, three-dimensional including foundational theory, clinical observations, and animal experiments and so on. Those trying to find a new structure of the meridian will have no results. Certainly, we do not doubt that meridian has it’s “fundamental substance”, but Dr. Jin Fenghan tried to use a “Feng Han Xiao Ti” to clarify the material basis of the meridian is unbelievable and it just like to fish in the air. Because we have enough justification to prove the “fundamental substance” is not one kind of substance. For example stroke, pain syndrome, psychiat-ric symptoms, skin herpes involve nerves and blood ves-sels, lymph nodes and other various physical infrastruc-tures.

We should recognize the understanding of ancients is simple, experienced and intuitionistic. Einstein [4] once pointed out that “the development of Western science is based on two great achievements that the Greek phi-losophers’ formal logic system (in Euclidean geometry), and the experimental system”. Chinese sages hadn’t taken this step.

Engels [5] also pointed out that, in Greece the forms of dialectical thinking are still natural simplicity. Because the Greeks have not mastered the way to analyses the nature, so they observe it as a whole. The contact of natural phenomena has not been proven in details, so the link of them is a direct and intuitive result. These analy-ses are also suitable for our Chinese. In the twenty-first century, we should make full use of logical analysis, positive and comprehensive study to unravel the mystery of the meridian. Meridian research not only belongs to TCM but also to the whole life sciences; not only to China but also to the whole world. So the meridian re-search is bound to progress with the development, and it also will benefit all the humanity. 6. References [1] G. J. Wang, “Current Situation of Meridian Research in

China,” Word Science Technology, Vol. 11, No. 4, 2009, pp. 532.

[2] L. X. Huang, “Outline of Chinese Acupuncture academic history,” Huaxia Publishing House Publishing House Press.

[3] Y. H. Du, “Four Kinds Spectrum of Disease introduced by Y. H. Du in Tianjin,” 2002. http://www.pharmnet.com. cn/cn/yyzx/2002/05/10/116295.html

[4] L. Y. Xu and D. N. Fan, “Einstein Anthology,” Commer-cial Press, Beijing, 1976.

[5] G. Y. Yu, “Natural Diagnostic Method, Engels,” People’s Publishing Agency Press, Beijing, 1984.



The Wu Xing Theory and Homeostatic Interaction of Organs

Yevgeny V. Albegov, Dmitry V. Butenko, Lyudmila N. Butenko CAD/CAE Systems Department; Volgograd State Technical University, Volgograd, Russia

E-mail: [email protected], [email protected] Received June 21, 2010; revised August 9, 2010; accepted August 13, 2010

Abstract In this paper the making of homeostatic models of the bioenergetic informational acupunctural human’s sys-tem of the “Pentacube” and the “Hexagon” according to the Wu Xing rules and energy shifts dynamics is considering. The making of the meridional homeostatic model of the bioenergetic informational acupunctural human’s system based on the existing knowledges about the human body meridians is considering. Prefer-ence to choice of the homeostatic functional models of the human body organs energies interaction is sub-stantiated. The meridional control submodel of homeostatic system is defined and constructed. The descrip-tion of control submodel, perspectives of application and development are represented. Keywords: System Stability, Homeostat, Homeostatic Model, “Pentacube”, “Hexagon”, Meridional Homeo-

static Model, Control Submodel, “Merkabah”

1. Introduction The fundamental issues of any system are the problems of stability. In this paper various types of stability and models, this stability is achieved with, are considering. As a prototype, we consider how to reach stability in complex systems, which are biological organisms.

For biosystems the stability issues is the problem of homeostasis [1]. As an example of this system class sta-bility, we selected the human organism and examined it based on the five-membered cycle of the Wu Xing which reflects the interaction of human organs groups.

In the process of studying of organs rhythm activity of the Yin-Yang cycles, we came to the conclusion that the interaction is homeostatic in nature. Then based on this conclusion we constructed the functional homeostatic “Pentacube” model. Next, on the basis of data available to us, the sixth group of organs of the “Fire' ” element was extracted. As a result we increased the order of the system stability transforming the “Pentacube” into the “Hexagon” model.

The next step of the model development was the ex-traction of regulating channels of processes and their switching centers in the homeostatic relationships. They are represented by the meridians of the human body, encephalon and spinal cord.

Based on the information about the levels of the system

organization processes, we have attempted to identify the implicit control structure of the meridional homeostatic system. On the basis of representation of two-dimensional discrete set of stars we have concluded that the control submodel is the “Merkabah” system.

The stellated tetrahedron is an invariant of the octahe-dron, and hence, it is quite a stable structure in different coordinate systems. The “Merkabah” submodel also proves the correctness of the hexagonal homeostatic model and the meridional homeostatic model: a stable system of biological type can be controlled only by a stable internal structure with the same stability level. In systems of the pentagonal type the correct stable internal model is not observed.

Thus, the stability of the human body functioning can be explained in terms of homeostatic nets. 2. Making of the Homeostatic Models

2.1. Making of the “Pentacube”

According to the Wu Xing theory, the pentagons of human organs Yang and Yin have two circular cycles. One is clockwise and stimulating in nature, the other is anticlockwise and depressive [2]. The pentagons’ stimulating and depressive processes can be graphically represented as a pentagram (a five-pointed star). This

Y. V. ALBEGOV ET AL.


46

may be due to the interaction between two poles of homeostats. Besides, if you combine the Yin and Yang organs relating to the same element according to the Wu Xing theory, you can get a number of Yin-Yang homeostats. The homeostatic nature of this interaction is confirmed by the fact that organs interactions in these homeostats are synchronized. For example, in homeo-stat of “Metal” element (lungs—large intestine) lungs become more active from 3 till 5 in the morning, and large intestine from 5 till 7 in the morning; in homeostat of “Water” element (kidneys—urinary bladder) urinary bladder becomes more active from 15 till 17 in the af-ternoon, and kidneys from 17 till 19 in the afternoon; in homeostat of “Wood” element (liver—gall bladder) gall bladder becomes more active from 23 till 1 in the night, and liver from 1 till 3 in the night; in the homeostat of “Fire” element (small intestine—heart), the heart be-comes more active from 11 till 13 in the afternoon, and small intestine from 13 till 15 in the afternoon; in the homeostat of “Earth” element (spleen/pancreas—stom-ach) stomach becomes more active from 7 till 9 in the morning, and spleen/pancreas from 9 till 11 in the morning. So, connecting all 5 known organs of Yin and Yang horizontally and vertically, we get the homeo-static functional “Pentacube”, in the vertexes of which there are human organs, and the edges are homeostatic connections between organs (Figure 1).

The direction of arrows on the figures of homeostatic models indicate “generating” (the green one) and “over-coming” (the black one) processes accepted and thor-oughly described in the Wu Xing theory.

In particular, alongside with “Pentacube” model one obtains a partial fractal model of the relationships hier-archy of homeostats. At the same time, the chain of “the conglutinated” homeostats represents the homeostat of higher organizational level. The new homeostat proc-esses information in the same manner as the component homeostats do but at qualitatively superior level [3]. 2.2. Making of the “Hexagon”

First of all, the premises for the existence of sixth group

Figure 1. Homeostatic “Pentacube”

of organs were made many hundreds years ago by an-cient Chinese scholars. In parallel with the system of five Zang-Fu, they created a system of circulation of the twelve meridians (Jing-Luo), the imaginary vessel, which circulates Qi energy, usually called “vital force”. After developing the system of meridians, the ancient Chinese scholars associated each meridian with a spe-cific internal organ. Meanwhile, onto the 12 meridians only 10 of the internal organs were assigned. So, the question if two more internal organs existed arose. For the additional Zang organ, the so-called “Master of the Heart” (Xin Zhu) also known as pericardium was se-lected. Onto the place of the sixth Fu organ, the “su-per-organ” was selected. It is known as San Jiao or triple heater. This organ of Three Jiaos is involved in meta-bolic processes of the organism. Currently, one can come across the fact that in addition to five-membered ring in the Wu Xing cycle, there is another equal “member”. The “Fire” element is dual and contains 4 elements (two Zang organs and two Fu organs) in that representation.

Secondly, although the element of “Fire” fills the hu-man body with vital energy, the physical-chemical me-chanism of oxidation (“internal warming”) in the “Fire” of heart—small intestine and in the “Fire” of pericar-dium—triple heater is different.

Thirdly, we turn to the stability of systems built on polygons. Hexagon has a higher symmetry axis L6 against the pentagonal L5. Structures of solids, characterized by the fifth-order axis of symmetry of the prohibited in classical crystallography, occur rarely in nature and have a prefix “quasi-”. The very nature aspires to create ob-jects of different complexity levels in the hexagonal structures. This property is manifested both at the mac-rocosm and microcosm levels.

Fourthly, the interaction rhythm (by analogy with the previously described homeostats) of organs such as peri-cardium—triple heater will also be homeostatic in nature: the pericardium becomes more active from 19 to 21 in the evening, and triple heater from 21 till 23 in the evening.

And fifth, in a model of five-membered cycle in pathological states of excess or lack of Qi energy in the organs, logical rhythmic of activity has a probabilistic nature, and points to an extreme degree of degeneration of the pentagonal system. In the model of six-membered cycle this uncertainty states is completely eliminated.

The ideas described above enable us to draw a conclu-sion that the sixth group of organs exists: triple heater— pericardium, relating to the element “Fire' ”. The proc-esses of stimulating and suppression and, accordingly, the homeostatic interactions of organs in sexangles of Yin and Yang of the homeostatic functional “Hexagon” are the same as the processes between the organs in the pentagons of “Pentacube” (Figure 2).



47

3. Making of the Meridional Homeostatic Model

Due to the fact, there are regulators in the compound of homeostatic bioenergetic system, then we suggest, that as channels of regulating the standard meridians are used, which have projections on the surfaces of human body. It is known, that besides 12 standard meridians, related to the definite organs of the human body, there are other subsys-tems of meridians, performing the functions of additional direct and feedback connections (“miraculous” meridians, tendinomuscular meridians, unpaired meridians, collat-erals and secondary meridians) [4]. To switching centers, transmitting control action into the unpaired posterome-dian and unpaired anteromedian pretend, to our mind, spinal cord for Yin organs and encephalon for Yang or-gans. Chief-regulator likely to represent consis- tent be-ginning of nature, pacing the works of homeostats of or-gans. Since inner homeostatic interactions between the organs in pentagon of organs are described as a five- point star (a star as a planar geometric figure is meant) and are exhaustive, so in sexangle of organs the inner homeostatic interactions will be described as a six-point star/hexagram, representing in form the projection of stellated trigonal bipyramid on the plane, and will be also exhaustive [5,6]. Consequently the conclusion can be made about the pres-ence of homeostatic connections between the homeostat of “Wood” element (liver—gall bladder) and homeostat of “Fire' ” element (pericardium—triple heater), and be-tween the homeostat of the “Fire” element (small intes-tine—heart) and the homeostat of “Earth” element (spleen/pancreas—stomach) in homeostatic “Hexagon” of the human body organs (Figure 3).

The given model of interaction of human organs ener-gies represents a hierarchy meridional and homeostatic net, where the meridional net represents a tracking sys-tem with feedback connection on the wholeness of ho-meostatic net system.

The preference of choosing forms of homeostatic models

Figure 2. Homeostatic “Hexagon”.

is based on “gold” polygons and polyhedrons since the structures having in its basis correct n-angles have the maximal degree of stability, information, self- organizing and a harmonicity, whether there are creations of civili-zations (the Egyptian pyramids, protective forts, calendars of ancient civilizations, the theory of “civilizing hexagon”, models of location of cities, models of classification of smells) or the very nature (structure of chemical sub-stances (fullerenes, frame structures), of a atomic nucleus, DNA chain, a crystal lattice and nanostructures) [7]. “The golden section” and corresponding “golden” properties of edges of functional homeostatic models are based on concepts of space of variability of natural processes and homeostat (elementary homeostatic system) [8].

4. Making of the Meridional Submodel

“Merkabah”

Proceeding from judgment that organizers and regulators of local processes in the control space of the lower level of system of correct polygons are stars (connected and not connected), it is possible to make a conclusion, that in homeostatic “Hexagon” (a polyhedron – a prism) of the meridional homeostatic system of energies interaction of human body organs such organizer-regulator of processes is a star-shaped polyhedron—Merkabah or a stellated tetrahedron.

Merkabah as a member of a row of Platonic solids (oc-tahedron is an invariant to stellated trigonal bipyramid) can model dualism. Merkabah as a process is the integral stage of biosystem genesis, namely biological-energy evolution of a cell: at the third stage of the ovicell division eight cells form a stellated tetrahedron and they are ab-solutely identical to each other [7]. Static cognitive model of interaction a stellated tetrahedron reflects the basic

Figure 3. Meridional homeostatic model.



48

rules of constructing of meridional biosystem and the technology of forming homeostatic connections as com-plete units. Knots in “Merkabah” system represent a brain as the switching centers of a human body. 4 knots of Yang-tetrahedron are the encephalon, and 4 knots of Yin-tetrahedron – the spinal cord. And as the switching centers encephalon and the spinal cord will be absolutely identical to character of controlling.

The key rule of constructing consists of the following: the meridian represents not unidirectional structural unit (brain – body), but a contour (brain – organ – brain), or a construction of the tunnel type with "two-way traffic". The technology of formation of homeostatic units is ob-viously displayed on a stellated trigonal bipyramid: ho-meostatic couple of Yin-Yang organs is formed owing to a crossing of meridians of the corresponding Yin-Yang organs. We can notice that such approach does not con-tradict, namely proves the correctness of meridional ho-meostatic model. Having connected homeostatic couples of Yin-Yang organs by a broken line in view of their timely activity, we receive a flat figure hexanema, de-termining a timely way [9] (Figure 4).

Proceeding the meridians in view of timely activity both in a stellated tetrahedron as a whole, and in Yin and Yang tetrahedrons, and consequently also a process of meridional energy circulation (signals), on the given cognitive models occurs at first clockwise, then changes its direction on counter, i.e. on an endless helix, and on a plane looks like an infinity sign.

Figure 4. Meridional submodel “Merkabah”.

5. Conclusions

The system approach to the construction of functional submodel allows to observe all scope of relations of “Merkabah” elements and connections between them and to approach to understanding of dialectic “plan” of the nature in organizing and controlling the processes, to see dynamics in statics. From the point of view of the applied component of the research, the given control submodel allows to understand the essence of pathological proc-esses in a human body, as infringement of energy flow (transferring signals) in meridional system with the fur-ther disbalance of rhythms of homeostats and displace ment of homeostatic energy from optimum areas to the risky areas. The same comprehension of occurrence of disease states in turn, on the basis of knowledge of tradi-tional Chinese medicine, will lead to forming and using of algorithms of updating of biosystem and taking out from pathological conditions by methods of not classical medicine, but phytotherapy or acupuncture. We’d like to note that “Merkabah” as conceptual model of relations of complicated compound elements, has all preconditions to deep system complication (by all aspects of the given concept), owing to the analysis and decomposition of components. 6. References [1] A. G. Teslinov, “Development of Control Systems:

Methodology and Conceptual Structures,” Globus, Moscow, 1998.

[2] W. Xing, 2005. http://en.wikipedia.org/wiki/Wu_Xing

[3] A. M. Stepanov, “The Fundamental Principle of the Medical Homeostatics,” MODEC, Voronezh, 1994.

[4] G. Luvsan, “Traditional and Modern Aspects of Oriental Acupuncture,” Nauka, Moscow, 1991.

[5] The Star (geometry), 2008. http://ru.wikipedia.org/wiki/ Звезда_(геометрия)

[6] The Bipyramid, 2008. http://ru.wikipedia.org/wiki/Бипирамида

[7] A. Stahov, A. Sluchenkova and I. Scherbakov, “The Da Vinci code and Fibonacci series,” Piter, SPb, 2007.

[8] Yu. M. Gorsky, A. M. Stepanov and A. G. Teslinov, “Homeostatic: Harmony in Game of Contradiction,” Reprotsentr A1, Irkutsk 1998.

[9] V. Eremeev, “A Diagram of Anthropocosmos,” ASM, Moskva, 1994.



The Effect of EDTA Chelation Therapy in Symptomatic Coronary Heart Disease: An Observational Study

Chulananda D. A. Goonasekera1, Rohini Tennakoon2, Premil N. Rajakrishna3, Gammadegedara A. Gunasena3, Chandima R. Wanniarachchi3, Asanka B. Yatawatta3, Udawatta A. D. D. Munidasa3

1 Department of Anesthesiology, Faculty of Medicine, Peradeniya 2Consultant Cardiologist, General Hospital, Kandy, Peradeniya

3Temporary Lecturer, Department of Anesthesiology, Faculty of Medicine, Peradeniya E-mail: [email protected]


Abstract Ethylene Diamine Tetra Acetic Acid (EDTA) chelation therapy has been considered a definitive alternative therapy for by-pass surgery in atherosclerotic cardiovascular disease for more than four decades. It is a rela-tively inexpensive method believed to restore blood flow in atherosclerotic vessels. However, the benefits of chelation therapy yet remain controversial in the treatment of ischemic heart disease. We observed the effect of EDTA chelation therapy on exercise tolerance in 13 volunteering patients receiving conventional treat-ment for established symptomatic coronary heart disease. Each patient received 30 weekly infusions of EDTA followed by monthly 12 boosters according to the ACAM protocol (American College for Advance-ment in Medicine). This was in addition to the conventional therapies they received from their respective physician in hospital. Stress ECG, echocardiography and coronary angiogram findings were obtained at the beginning of treatment. The distance that a patient could walk on level ground at moderate speed and the number of steps he/she can climb up on a staircase until he/she begins to feel either chest pain or breathless-ness were the two clinical parameters of exercise tolerance recorded to grade angina. Liver and renal func-tions were tested at 1st, 5th, 10th, 15th and 30th infusions. Of the 13 patients, 11 showed improvement in angina grading whilst 2 experienced no effect. One patient improved from angina grade IV to I, 6 from grade III to I, 1 from grade III to II and 3 from grade II to I. A statistically significant reduction in the mean score (p = 0.002) was noticed at 6th month of treatment when compared to that of the first month. A significant 1.7 fold increase (p = 0.009) in the mean SGPT level was observed at the 30th infusion when compared to the pre-treatment values. The SGOT level showed no significant change (p = 0.664). None of the patients showed clinical features of hepato-cellular damage. The mean serum creatinine level showed a trend for re-duction (p = 0.083) with treatment. The recognized side effects of intravenous EDTA chelation therapy such as liver damage, renal damage, hypersensitivity, symptomatic hypocalcaemia, and thrombophlebitis were not encountered. Thus, EDTA chelation therapy as prescribed by the ACAM protocol seems safe and effective in improving exercise tolerance in ischemic heart disease when administered concurrently with conventional therapy. Keywords: EDTA Chelation Therapy, Coronary Heart Disease, Exercise Tolerance

1. Introduction

Ischaemic heart disease (IHD) resulting from the nar-rowing of coronary arteries due to atheromatous plaques, is the commonest cause of deaths in Sri Lanka [1]. Its definitive treatment comprise of revascularization sur-gery [i.e. Percutaneous Transluminal Coronary Angio-

plasty (PTCA), or Coronary Artery Bypass Grafting (CABG)]. Its symptomatic management includes drug therapy (aspirin, beta blockers, nitrates, etc.) and lifestyle modifications. Surgical revascularization procedures are costly and not widely available. Often, most needy pa-tients are unsuitable for surgery due to their poor physi-cal condition or age. In this context, the search for an

C. D. GOONASEKERA ET AL.


50

alternative, definitive and cost effective treatment is im-portant. One such treatment is “chelation therapy” i.e. a therapy of intravenous Ethylene Diamine Tetra Acetic Acid (EDTA), a synthetic amino acid.

The benefits of EDTA Chelation therapy practiced worldwide since 1950s are still unclear. This is because of scarcity of well-controlled clinical trials. The exact mechanism of action of EDTA is not yet fully under-stood. One theory suggests that EDTA might work by removing calcium from atheromatous plaques clearing the blocked coronary arteries. Others believe that EDTA reduces oxidative stress related damage on coronary vessels [2]. Some evidence suggests that EDTA reduces platelet aggregation [3].

In 2001, a metanalysis of 19 studies with data on 22765 patients revealed a statistical correlation coeffi-cient of 0.88, which was indicative of a positive rela-tionship between EDTA therapy and improved cardio-vascular function [4]. There were also a few studies such as the Calgary PATCH EDTA chelation therapy study 2002, which stated that there was no evidence to support chelation therapy in patients with ischaemic heart disease, stable angina, and positive treadmill test for ischemia [5]. This study has been criticized as non scientific [6].

In spite of above controversies, “Chelation Therapy” for ischaemic heart disease is practiced in many coun-tries including USA, UK, Australia and New Zealand using accepted protocols (ACAM, Rozema TC). An overview of all clinical investigations of chelation ther-apy conducted in the year 2000 states that out of 22 uncontrolled studies, 20 reported subjective sympto-matic improvement [7]. A study of 18 patients using Technetium 99m echocardiography has shown a statis-tically significant improvement in left ventricular ejec-tion fraction with chelation therapy [8]. Several recent reviews conducted on the subject, whilst acknowledg-ing the fact that there is a scarcity of solid data to ap-prove its use [9], conclude that EDTA therapy may be used in the context of a research trial [10] or as a ‘last resort’ cardiovascular revitalization strategy extending beyond revascularization [11] especially in patients considered unsuitable for revascularization [12]. On the other hand EDTA therapy as prescribed is not consid-ered a highly invasive or harmful therapy [13]. As a result groups of physicians in Sri Lanka conduct chela-tion therapy as an ‘alternative medicine’ for CABG in ischaemic heart disease, on a non-profit basis, for vol-unteering patients.

This paper presents an observational study of out-comes, adverse effects, exercise tolerance and hepato renal function in 13 such patients who have completed a standard course of EDTA chelation therapy over the past three years at a non-profit clinic conducted in the Central

Province of Sri Lanka. 2. Methods Volunteering patients, self referring themselves for che-lation therapy whilst receiving conventional drug treat-ment for established ischaemic heart disease qualified for recruitment for this study. Subjects with co-morbid con-ditions such as a) psychiatric illness affecting under-standing b) severe renal impairment (s. creatinine > 2.5mg/dl) c) significant liver disease (liver enzymes> twice the upper limit) d) severe cardiac failure e) allergy to EDTA f) suspected lead encephalopathy or else if pregnant were excluded.

Informed written consent was obtained after explain-ing the background, the ambiguous nature of treatment, and side effects. Their pre-treatment assessment included a history, physical examination and baseline investiga-tions: i.e. urine for sugar and protein, serum creatinine, full blood count, SGOT, SGPT, lipid profile, fasting blood sugar, ECG, 2D Echocardiography, and treadmill exercise test (modified Bruce Protocol). All angina pa-tients were graded according to the Canadian Cardio-vascular Society Classification System before the treat-ment, namely, Class I—No limitation of physical activity (Ordinary physical activity does not cause symptoms.), Class II—Slight limitation of physical activity (Ordinary physical activity does cause symptoms.), Class III— Moderate limitation of activity (Patient is comfortable at rest, but less than ordinary activities cause symptoms.), and Class IV—Unable to perform any physical activity without discomfort, therefore severe limitation (Patient may be symptomatic even at rest) [14].

ACAM Protocol for the safe and effective administra-tion of EDTA Chelation therapy was followed [8]. Each patient was given weekly 30 infusions followed by monthly 12 boosters. The duration of therapy was ap-proximately 1½ years. The therapeutic infusions were given over a period of 3 hours according to the following formula.

The EDTA dose, rate of infusion and the frequency of administration were re-adjusted according to the creatinine clearance. If the creatinine clearance was 100 ml/ min/1.73m2, full dose of EDTA was given. If it was be-low 100 ml/min, for example 70 ml/min/1.73m2, then 70% of the EDTA dose was given. The total fluid load was reduced to 400 ml or 300 ml in patients presenting with clinical features of heart failure (lung basal crepita-tions, ankle oedema, etc.) [8].

Patients were advised to take an adequate meal prior to the infusion. The infusion was given in a comfortable sitting position and patients were served with refresh-ments and drinking water if required.



51

Investigations were repeated soon after the 5th, 10th, 15th and 30th weekly infusions to assess their renal and liver functions. Their lipid profiles were checked at 10th and 30th infusions. The distance that a patient could walk on level ground at moderate speed and the number of steps he/she can climb up on a staircase until he/she be-gins to feel either chest pain or breathlessness were re-corded prior to, and at the end of the first course (30 in-fusions) of EDTA therapy.

The importance of life style adjustment measures was emphasized. Regular aerobic exercise, quitting smoking and alcohol, stress free life, and dietary control measures such as limitation of salt, sugar, and fat intake and in-crease in fiber content were the key advices conveyed to the patients via leaflets and interviews.

Angina grading was reassessed at the end of 30 weekly infusions using two subjective parameters; namely a) the distance that a patient could walk on level ground at moderate speed and b) the number of steps he could climb up on a staircase until he begins to feel ei-ther chest pain or breathlessness. The patient’s opinion on the effectiveness of the treatment was also sought. The biochemical parameters; Urine sugar and protein, Serum creatinine, Full blood count, and SGOT, SGPT, and Lipid profile were repeated.

All causes of death, myocardial infarction, stroke, hospitalization for angina, and coronary revasculariza-tion surgery were taken as study end points.

3. Results

All 13 patients (10 male, age median 57yrs, range 45-71) completed 30 infusions. Of the 13, 10 patients had had coronary angiograms and 9 had been recommended to have CABG. Their angina grading before and after treatment is shown in Table 1.

Of those 9 patients, 7 improved in angina grading. One patient recommended for medical management also im-proved.

Of 11 patients with stress ECGs, 7 were positive at stage I, 2 at stage II, and 2 at stage III. All patients im-proved in angina grading except for the 2 patients who were positive at stage I.

The mean blood pressures of our study group during treatment are shown in Figures 1 & 2. The reduction in the mean systolic blood pressure at 3rd, 4th, 5th and 6th months were statistically significant when compared to that of the first month.

The patient’s opinion on the effectiveness of treatment obtained after completion of the 30 infusions is shown in Table 2. The number of steps they could walk on flat ground and climb significantly improved as shown in Figure 3 and Figure 4 respectively.

Table 1. Angina grading before and after EDTA therapy course i.e. 30 infusions.

Angina Grading

Before infusions After 30 infusions N=

IV I 1

III I 6

III II 1

III III 1

II I 3

II II 1

Table 2. Patients’ perception on the effectives of the treat-ment.

Patients’ opinion Number of patients

Symptoms totally improved 2

Symptoms improved to a greater extent 7

Symptoms improved to a lesser extent 2

No change 2

Symptoms worsened 0

Month

7654321

95%

CI

Syst

olic

Blo

od P

ress

ure

(mm

Hg)

150

145

140

135

130

125

120

115

110

Figure 1. Monthly systolic blood pressure during chelation therapy. The mean SGPT levels before and after treatment were 29.19 U/L and 41.58 U/L and showed a significant 1.42 fold rise (p = 0.009). The mean SGOT was 27.64 U/L and 32.64U/L showed no significant change (p = 0.664). The mean serum creatinine level was 1.10mg/dl before and 1.02 mg/dl after treatment and showed a declining trend (p = 0.083).

Major complications of EDTA i.e. liver damage, renal damage, hypersensitivity, symptomatic hypocalcaemia and thrombophlebitis were not seen in our patients. One patient developed faintishness after her 2nd infusion. An-other patient quit the treatment at the 5th infusion, com-plaining that a rash in the leg exacerbated with the infusion.



52

Month

7654321

95%

CI

Dia

stol

ic B

lood

Pre

ssur

e

90

85

80

75

70

65

60

Figure 2. Monthly diastolic blood pressure during chelation therapy.

Figure 3. The number of steps subjects could walk on flat ground before and after the completion of full course of EDTA. 4. Discussion We studied the effect of EDTA therapy in 13 patients with established symptomatic ischaemic heart disease who were on conventional drug treatment with life style modifications. They were also either considered not suitable for cardiac revascularization or were not willing to undergo the procedure. Of the 13 patients, 11 showed an improvement in angina grading at least by one class. Their exercise tolerance as indicated by walking distance and climbing ability substantially improved. Seven

Figure 4. The number of steps subjects could climb before and after the completion of full course of EDTA patients who had been recommended for early CABG were within the improved group.

Objective parameters to assess the effect of EDTA, such as stress ECG for exercise tolerance, echocardi-ography for cardiac function were not used in this study as patients could not afford the expenses.

The mean blood pressure of our study group was within the normal range. Interestingly we found a sig-nificant reduction in systolic blood pressure between 8th and 24th week of EDTA infusion therapy. Most of these patients were on antihypertensive medication and regular physical exercise, which are known to control blood pressure. It has also been previously reported that chela-tion therapy lowered the systolic blood pressure at all stages of the sub maximal treadmill stress test in both exercising and non exercising groups [15]. Therefore, it is worth considering further research to elucidate whether chelation therapy has a permissive action on controlling hypertension.

Critics of chelation therapy have frequently suggested that reported improvements are a placebo effect. A pla-cebo effect begins shortly after its first administration and rarely, persists for more than three months. Benefits shown in our study are 30 weeks apart. Chelation therapy, by contrast, shows its full range of benefits quite slowly. Usually it required not only several months of therapy but also an additional several months after the course of therapy for the full benefit of treatment to occur. The benefits accrued generally persist for years thereafter [16].

The results we obtained might have been affected by other confounding factors such as vitamin B supplemen-



53

tation, life style modifications and conventional drug therapy. Several studies have shown that favorable life style modifications like quitting smoking, regular exer-cise and dietary adjustments have a significant effect on attenuating abnormal cardiovascular risk factors [17].

Some authors continue to refuse EDTA Chelation therapy claiming that it has serious side effects including renal and hepatic toxicity. EDTA does have them when given at higher doses for heavy metal poisoning. So would it be for any other medication if toxic doses are used [2]. However, with the use of accepted protocols (e.g. ACAM) for EDTA therapy in heart disease none of the major side effects have been reported in previous studies. Out observations are similar.

SGPT levels were within the normal range before treatment and only two patients had values slightly above the upper normal at the end of treatment (46.4 U/L and 57 U/L; normal range 0-40 U/L). One patient who had elevated SGOT level at the end of treatment (99 U/L) also had an abnormal level (47 U/L) pre-EDTA treat-ment. However these post treatment liver enzyme eleva-tions are relatively low when compared to that of known liver pathologies [18]. Since the study group was on statins (a group of cholesterol lowering drugs known to raise liver enzymes) as a component of their conven-tional therapy, a reliable causal relationship between EDTA and raised liver enzymes could not be made. However, none of our patients showed any clinical fea-tures of hepato-cellular damage during the study period.

We observed a decreasing trend in the mean serum creatinine level during treatment. EDTA induced renal damage is a very rare adverse effect and it is usually as-sociated with higher doses [19]. There is even a sugges-tion that this treatment procedure may improve renal function. A study carried out on 383 subjects with chronic degenerative disorders who were treated with EDTA for 50 days showed an overall decline in fasting serum creatinine levels [20].

Other known side effects of intravenous EDTA chela-tion therapy (hypersensitivity, symptomatic hypocalcae-mia and thrombophlebitis) were not encountered during our study period. Intravenous access related complica-tions were minor and rarely observed. Pain at the injec-tion site and allergy to plastic tourniquet (mild erythema) troubled a rare patient.

5. Conclusions

We observe that EDTA chelation therapy is effective in improving the clinical outcome in a majority of IHD pa-tients with negligible side effects when given according to approved protocols. Further research is needed to con-firm this causal relationship. The variability of the effect

of EDTA on different patients creates the need for fur-ther research to recognize the clinical subset of IHD pa-tients for whom it is most suitable.

6. Acknowledgements

We thank all staff members of EDTA chelation therapy clinic, Kandy, and all patients who participated. We ap-preciate the support received from the Torance Company Pharmaceuticals to import the relevant drugs under a special license. 7. References [1] World Health Organization, “Mortality Country Fact Sheet,”

2006. http://www.who.int/whosis/mort/profiles/mort_searo_ lka_srilanka.pdf

[2] NCCAM, National Institutes of Health, “Questions and Answers: The NIH Trial of EDTA Chelation Therapy for Coronary Artery Disease,” 2004. http://nccam.nih.gov/ health/chelation/q-and-a.htm

[3] G. Kindness and J. P. Frackleton, “Effect of Ethylene Diamine Tetra Acetic Acid (EDTA) and Platelet Aggre-gation in Human Blood,” Journal of Advancement in Medicine, Vol. 2, No. 4, 1989, pp. 519-530.

[4] L. Chappell and J. P. Stahl, “The Correlation between EDTA Chelation Therapy and Improvement in Cardio-vascular Function: A Meta Analysis,” Journal of Ad-vancement in Medicine, Vol. 6, No. 3, 1993, pp. 139-160.

[5] M. L. Knudtson, D. G. Wyse, P. D. Galbraith, R. Brant, K. Hildebrand, D. Paterson and D. Richardson, “Chela-tion Therapy for Ischemic Heart Disease: A Randomized Controlled Trial,” Journal of the American Medical As-sociation, Vol. 287, No. 4, 23-30 January 2002, pp. 481- 486.

[6] “Critique of the Calgary PATCH EDTA Chelation Study,” Adapted from a report by Croft Woodruff in Canada, 2007. http://www.drcranton.com/calgarystudy.htm

[7] E. Ernst, “Chelation Therapy for Coronary Heart Disease: An Overview of All Clinical Investigations,” American Heart Journal, Vol. 140, No. 1, July 2000, pp. 139-141.

[8] E. M. Cranton, “Protocol of the American College of Advancement in Medicine for the Safe and Effective Administration of EDTA Chelation Therapy”. Journal of Advancement in Medicine, Vol. 2, No. 1-2, 1989, pp 269- 305.

[9] E. Ernst, “Chelation Therapy for Coronary Heart Disease: An Overview of All Clinical Investigations,” American Heart Journal, Vol. 140, No. 1, July 2000, pp. 139-141.

[10] J. S. Shrihari, A. Roy, D. Prabhakaran and K. S. Reddy, “Role of EDTA Chelation Therapy in Cardiovascular Diseases,” National Medical Journal of India, Vol. 19, No. 1, January-February 2006, pp. 24-26.

[11] P. M. Kidd, “Integrative Cardiac Revitalization: Bypass Surgery, Angioplasty, and Chelation. Benefits, Risks, and Limitations,” Alternative Medicine Review, Vol. 3, No. 1,



54

February 1998, pp. 4-17.

[12] H. Quan, W. A. Ghali, M. J. Verhoef, C. M. Norris, P. D. Galbraith and M. L. Knudtson, “Use of Chelation Ther-apy after Coronary Angiography,” American Journal of Medicine, Vol. 111, No. 9, 15 December 2001, pp. 686- 691.

[13] D. M. Seely, P. Wu and E. J. Mills, “EDTA Chelation Therapy for Cardiovascular Disease: A Systematic Re-view,” BMC Cardiovascular Disorders, Vol. 5, 2005, p. 32.

[14] L. Campeau, “Grading of Angina Pectoris,” Circulation, Vol. 54, No. 3, September 1976, pp. 522-523.

[15] E. W. McDonagh, C. J. Rudolph, E. Cheraskin and D. G. Wussow, “The Effect of EDTA Chelation and Supportive Multivitamin/Trace Mineral Supplementation with and without Physical Activity upon Systolic Blood Pressure” Journal of Orthomolecular Psychiatry, Vol. 13, 1984, pp. 1-9.

[16] E. M. Cranton and J. P. Frackelton, “Current Status of

EDTA Chelation Therapy in Occlusive Arterial Disease,” 2001. http://www.drcranton.com/chelation/emcjpf.htm

[17] R. Yamamoto, T. Kawamura, K. Wakai, Y. Ichihara, T. Anno and Y. Mizuno, “Favorable Life-Style Modification and Attenuation of Cardiovascular Risk Factors,” Japa-nese Circulation Society, Vol. 63, No. 3, March 1999, pp. 184-188.

[18] “EDTA Chelation: The Real ‘Miracle’ Therapy for Vascu-lar Diseases,” Journal of Life Enhancement, 1997. http:// www.life-enhancement.com/article_template.asp?ID=78

[19] H. R. Casdorph, “EDTA Chelation Therapy, Efficacy in Arteriosclerotic Heart Disease,” Journal of Holistic Medi-cine, Vol. 3, No. 1, 1981, pp. 53-59.

[20] M. Elmer and E. M. Cranton, “Protocol of the American College for Advancement in Medicine for the Safe and Effective Administration of EDTA Chelation Therapy,” Journal of Advancement in Medicine, Vol. 2, No. 1-2, 1989, pp. 269-303.



Prevalence of Glucose-6-Phosphate Dhydrogenase Deficient Neonates and Cost Effective of Enzyme Screening

in Tehran, Iran

Hossein Dahifar1, Aboulfazl Ghorbani2, Manijeh Ghods3 1Associate Professor of Shaheed Beheshti University Medical Sciences and Health Services

Shohada Hospital, Pediatric-Department, Tajrish Sq, Tehran, Iran 2Booali Hospital, Pathology Department, Marivan, Iran

3Medical microbiology, Sandwell and West Birmingham Hospital, Birmingham, England E-mail: {dr-dahifar, dr_ghorbani 48573}@yahoo.com, [email protected]


Abstract Background: Newborn screening is an area with potential for immense impact in lifelong morbidity and mortality. Objective: To determine cost effective for glucose-6-phosphate dehydrogenase (G-6-PD) enzyme screening and prevalence of deficiency in newborns and children in Tehran, Iran. Materials and Methods: All full term newborns, neonates with icterus and children with acute hemolysis were evaluated to determine prevalence of G-6-PD enzyme deficiency and cost effectiveness of a screening test for G-6-PD enzyme defi-ciency. The qualitative color reduction test performed on healthy newborns and quantitative test on newborns with icterus and children with acute hemolysis. Results: Three (2%) of 146 screened newborns were G-6-PD deficient at a cost of $ 18 for three positive screening tests and $ 856 for 143 unaffected newborns Thirty-one (11.4%) of 272 newborns readmitted with icterus were G-6-PD deficient with a cost of 31 tests were $186 and $1446 for unaffected newborns. The only significant difference between G-6-PD deficient and normal newborns was bilirubin level (p < 0.001). Eleven (0.2%) of 5054 hospitalized children were found to be G-6-PD deficient at a screening cost of $ 66 and $ 3.258 for remainder children. The prevalence of G-6-PD is estimated to be approximately 2.4% and 2.2% in males and females respectively. Conclusion: Glucose-6- phosphate dehydrogenase screening in newborns is not cost effective and for prevention of hemolysis during the next years all newborns admitted with icterus should be evaluated for G-6-PD enzyme deficiency. Keywords: Screening, Newborns, G-6-PD

1. Introduction

Mass newborn screening is the second type of prospec-tive care. It is a very important issue for all physicians caring for neonates because it combines a number of significant medical and legal issues. One of the medical requirements of an acceptable mass screening program for a specific disease is that the test should be simple and inexpensive [1]. One of the accepted but less widely practiced screening test is for G-6-PD enzyme deficiency recommended by WHO expert group [2]. In addition, recommendation of WHO is all newborns should be car-ried out screening in area with a prevalence of G 6 PD deficiency of 3-5% or more in males.[3] In the United States, there are significant state-to-state differences in the disorders that are included in initial newborn screen-

ing, the methods of screening, and follow up [4]. Distribution of the G-6-PD deficiency varies among

different population reflecting geographic and ethnic variations. Each society is unique and must make its own choices in neonatal screening based on its economy, epidemiology and ethnics. We have decided to determine prevalence of G-6-PD enzyme deficiency and whether or not G-6-PD screening in newborns is economically ef-fective. 2. Materials and Methods

This prospective study was conducted in Tehran, Iran, at Aria Hospital and Tehran Children Hospital, including neonates born in Aria Hospital from April 2006 through March 2007. After obtaining the informed consent from

H. DAHIFAR ET AL.


56

one parent enrolled neonates were screened for G-6-PD enzyme level. Similarly enrolled newborns with icterus and hospitalized children with acute hemolisis were screened.

The qualitative color reduction G-6-PD screening test (Kit Saba Laboratory (Tehran, Iran)) was performed on all healthy newborns before hospital discharge. Results were reported as normal or deficient. The quantitative test (kit chem. Enzyme) was performed on children with acute hemolysis and newborns with icterus with results reported in IU/gHb (normal range 7.5-14.5 IU/gHb). We economically compared morbidity (rate of readmission of newborns with jaundice) and children with symptoms of pallor, dark urine, icterus, abdominal pain (acute hemolysis) with G-6-PD screening test results of new-borns.

3. Results

During the entire study period, a total of 146 (33.8%) of 431 uncomplicated full term or near term newborns (82 males) with a mean birth weight of 3278.4 g born at Aria hospital were screened for G-6-PD enzyme. Of these, 3(2%) that all were male newborn were G-6-PD deficient at a rate of 20 per 1000 live newborns. The cost of G-6-PD for every one newborn is about $6 or $6000 per 1000 newborns. A total of 5054 children with different chief complaints and ages were hospitalized at Tehran Children Hospital over the period, of these, 11(0.2%) or 2 per 1000 hospitalized children with icterus, pallor, dark urine and abdominal pain (acute hemolysis) had con-sumed fava beans within two days. Of these, 7(63.6%) of 11 children were males. The mean first blood hemoglo-bin level was 7.8g% and G-6-PD test revealed deficiency. Eight (72.7%) of 11 children were transfused and re-mainder received supportive therapy and all were dis-charged after two nights. The cost of two nights hospi-talization for one patient with parent is about $140 or $1540 for 11 patients. Screening of 5054 patients cost $30324. The cost of hospitalization of 2 per 1000 chil-dren with G-6-PD deficiency is $280 and the cost of screening 1000 children is $6000. The comparison of cost of 1000 G-6-PD screening with two deficient per 1000 children is very expensive and is not economical. On the other hand, 31(11.4%) of 272 icteric newborns (13 males) who were admitted over the same period were G-6-PD deficient and 6 (19.3%) of 31 newborns (3 fe-males) recieved exchange transfusion. The prevalence of G-6-PD deficiency is approximately 2.4% and 2.2% in male and female newbornbs respectively and characteris-tics of newborns are shown in Table 1. The only signifi-cant difference between normal and deficient G-6-PD newborns is bilirubin level (p < 0.001). There is no sta-tistically significant difference between exchange trans-

Table 1. Characteristics and laboratory examination of newborns with jaundice.

Normal G-6-PD

G-6-PD deficient

Normal G-6-PD

Exchange transfused

G-6-PD deficient

Exchange transfused

Total No 236 13 31 6

Males 121 6 18 3

Females 115 7 13 3

Age / day 8.6 5.7 6.4 4.9

Hemoglobin g%

14 15.9 15.4 17.4

Reticulo-cytes%

0.9 1.7 1.2 1.2

Bilirubin mg%

15.9 27 21.6 28.4

The numbers are defined as mean

fused newborns with normal G-6-PD and deficient new-borns. 4. Discussion In the current study we showed cost of G-6-PD screening in uncomplicated healthy full term newborns. In icteric neonates associated with G-6-PD deficiency their jaun-dice can not be related only to G-6-PD deficiency but confounding factors such as breast milk, or poor nursing technique and perinatal factors such as maternal diabetes, induced or augumented labor, superimposed on G-6-PD deficiency may exacerbate hyperbilirubinemia, treatment costs, duration of hospitalization and need for exchange transfusion.

Clearly, the fact that a G-6-PD deficient infant also develops hyperbilirubinemia or kernicterus does not mean that G-6-PD deficiency was the primary cause of the severe hyperbilirubinemia [5]. In our study such con-founders can-not be defined and we can not attribute all hyperbilirubinemia to G-6-PD deficiency. In our study, G-6-PD deficiency in newborns or children does not in-duce morbidity or mortality such as mental retardation or other irreversible clinical manifestation. The most com-mon clinical manifestations of G-6-PD deficiency is neonatal jaundice and acute hemolytic anemia in children related to medications such as Chinese remedies, naph-thalene, henna, fava and oxidant drugs particularly in Iran, henna, feva bean, naphthalene, aspirin and infection. The only clinical manifestation in our newborns with G-6-PD deficiency was bilirubin level higher than nor-mal G-6-PD newborns Table 1. Routine screening of all newborns will not identify every affected G-6-PD defi-cient newborns. The male G-6-PD deficient infants will be detected, but many heterozygot females will be missed [6]. In our study, all newborns screened for

H. DAHIFAR ET AL.


57

G-6-PD deficiency were males. There were not differ-ences between hemoglobin level and number of reticu-locytes on automated differential in G-6-PD deficient and normal G-6-PD newborns. We do not have evidence of hemolysis in newborns. Although it has previously been shown that G-6-PD deficiency can induce kernic-terus as a complication of G-6-PD deficiency associated neonatal jaundice in many population groups [7] our study did not demonstrate any kernicterus in newborns with jaundice. Our study demonstrated prevalence of G-6-PD deficiency approximately 2.4% in males and it is versus of recommendation of WHO [3]. Given that in this study only 0.2% of young children were admitted with acute hemolysis from fava bean ingestion. If we compare them with children due to gastroenteritis with prevalence of approximately 41% is extremely low [8]. In comparison the cost of newborn screening is also low. Currently, to our knowledge, newborn G-6-PD screening is not performed in the other provinces of Iran and the prevalence of G-6-PD deficiency in north, south, east and west provinces should be different and I hope to be determined by other investigators. 5. Conclusions This study demonstrated that global G-6-PD enzyme screening in newborns in Tehran, Iran for prevention of hemolysis following ingestion some drugs, fava beans or simultaneous with infection during the next years is not economically feasible. We suggest all newborns with jaundice should be evaluated for G-6-PD enzyme defi-ciency and that prevalence of deficiency in all provinces of Iran should be determined before establishing a uni-versal Iranian G-6-PD deficiency screening program.

6. Acknowledgements We thank Mrs Manijeh Hejazi and Miss Mojgan Moosavi

for their assistances.

7. References [1] A. A. Fanarof and R. J. Martin, “Neonatal-Perinatal

Medicine Disease of the fetus and infant,” 5th Edition, Mosby, USA, 1992.

[2] A. Larsson, “Neonatal Screening for Metabolic, Endo-crine, Infectious and Genetic Disorders,” Clinic in Peri-natology, 2001, Vol. 28, No. 2, pp. 449-461.

[3] WHO Working Group, “Glucose-6-Phosphate Dehydro-genase Deficiency,” Bulletin of the Health Organization, Vol. 67, 1989, pp. 601-611.

[4] B. K. Burton, “Inherited metabolic disorders,” In: G. B. Avery, M. A. Fletcher and M. G. MacDonald, Neonatol-ogy, Pathophysiology and Management of the Newborn, 5th Edition, Lippincott Williams and Wilkins, USA, 1999, pp. 821-837.

[5] M. Kaplan and C. Hammerman, “Glucose-6-Phosphate Dehydrogenase-Deficient Neonates: A Potential Cause for Concern in North America,” Pediatrics, Vol. 106, No. 6, 2000, pp. 1478-1480.

[6] M. Kaplan, E. Beutler, H. J. Vreman, C. Hammerman, E. Levy-Lahad, P. Renbaum, et al., “Neonatal Hyperbiliru- Binemia in Glucose-6-Phosphate Dehydrogenase Defi-cient Heterozygotes,” Pediatrics, Vol. 104, No. 1, Part 1, 1999, pp. 68-74.

[7] M. G. McDonald, “Hidden Risks: Early Discharge and Bilirubin Toxicity Due to Glucose-6-Phosphate Dehydro-genase Deficiency,” Pediatrics, Vol. 96, No. 4, Part 1, 1995, pp. 734-738.

[8] H. Dahifar, A. Ghorbani and M. Ghods, “Isolated and Non-Isolated Enteric Pathogens in Children with Diar-rhea and Related Laboratory Characteristics,” MHIRI, Vol. 22, 2008, pp. 40-46.

Documents

CM 1.2 contents · Chinese Medicine (CM) Journal Information SUBSCRIPTIONS The Chinese Medicine (Online at Scientific Research Publishing, ) is published quarterly by Scientific Research