[Effect of Negative Emotions on Serum Levels of Adrenocorticotropic Hormones and Neuropeptide Y in Hepatitis B Liver Cirrhosis Patients].

OBJECTIVE: To explore the effect of negative emotions on serum levels of adrenocorticotropic hormone (ACTH) and neuropeptide Y (NYP) in hepatitis B liver cirrhosis (HBLC) patients. METHODS: Totally 617 HBLC patients were assigned to the negative emotion group (415 cases) and the non-negative emotion group (202 cases) judged by negative emotions. Case numbers of various grading Child-Pugh were recorded in the two groups. Their liver functions were compared between the two groups. Serum levels of ACTH and NPY were detected using double antibody sandwich enzyme-linked immunosorbent assay (ELISA) in the two groups. RESULTS: There was no statistical difference in Child-Pugh grading between the two groups (χ2 = 0.65, P = 0.72). Compared with the non-negative emotional group, serum ACTH levels decreased significantly in the negative emotion group with statistical difference (P < 0.05). There was no statistical difference in serum ACTH levels between the two groups (P > 0.05). CONCLUSION: The negative emotion of HBLC patients was not related to the serum ACTH level, but to relatively lower-concentration serum NPY levels.

Intervention Effect of Baduanjin Exercise on the Fatigue State in People with Fatigue-Predominant Subhealth: A Cohort Study.

OBJECTIVES: To observe the effect of a Baduanjin exercise intervention on fatigue in people with fatigue-predominant subhealth (FPSH). PARTICIPANTS: A total of 131 participants were included in this cohort study. Those who met the inclusion criteria were grouped into the Baduanjin cohort (n=64) or the control cohort (n=67). INTERVENTION: The Baduanjin cohort exercised for 30 minutes twice a day for 6 weeks; the control cohort received no treatments. The study period was 18 weeks. OUTCOME MEASURES: The fatigue states of all participants were evaluated by using the Fatigue Self-Assessment Scale (FSAS) at baseline, the fourth and sixth weeks of treatment, and during post-treatment follow-ups at the end of the 12th and 18th weeks. RESULTS: The scores for the six factors on the FSAS at baseline did not significantly differ between the two groups. In the Baduanjin cohort, the total score on the FSAS (TSF) and scores for physical fatigue (PF), mental fatigue (MF), consequence of fatigue (CF), and response of fatigue to sleep and rest (RFSAR) were significantly decreased at the end of the sixth week. These patients also had significantly decreased scores for TSF, PF, MF, and CF at baseline, at the end of the fourth and sixth weeks of the intervention, and during post-treatment follow-ups at the end of the 12th and 18th weeks. No adverse effects of treatment were reported. CONCLUSIONS: This study suggests that Baduanjin exercise has an effect on relieving fatigue in patients with FPSH.

Distribution characteristics of meridian sinew (jingjin) syndrome in 313 cases of whiplash-associated disorders.

OBJECTIVE: To investigate and analyze the characteristics of Meridian Sinew (Jingjin) syndrome in patients with whiplash-associated disorders (WAD). METHODS: From August 2010 to September 2011, 313 WAD cases from New York and California states were collected. The survey mostly collects the information of "Sinew Knotted Points" and symptoms of four types of Meridian Sinew differentiation-Taiyang, Shaoyin, Shaoyang and Yangming. RESULTS: Among the cases which are on the average of medium injury level, the higher frequency of "Sinew Knotted Points" tenderness were found on Jianwaishu (SI 14), Jianzhongshu (SI 15), Tianchuang (SI 16), C3-6 Spinous Process, Dazhui (GV 14), Fengchi (GB 20), Tianliao (SJ 15) and Tianding (LI 17). The most commonly presented symptoms were widespread spasm and tenderness in the neck (Taiyang), difficulty in lateral flexion (Shaoyang), problems of extension and flexion (Taiyang), and stiffness and pain during neck movement (Yangming). Among the cases, 237 cases (75.72%) were related to Taiyang Meridian Sinew syndrome, 82 cases (26.20%) to Shaoyin syndrome and 175 (55.91%) and 176 (56.23%) cases to Shaoyang and Yangming syndrome respectively. The most of cases presented in a combination format. The syndrome distribution under Grade I, II and III reflected that more combination of the Meridian Sinew syndromes in the whiplash injury patients which is resulted from more severity of injury. CONCLUSION: It is practical to identify the location of abnormality through Meridian Sinew differentiation, considering both "Sinew Knotted Points" tenderness and corresponding symptoms, for the local neck symptoms of WAD.

Xiaopiyishen Herbal Extract Granule Improves the Quality of Life among People with Fatigue-Predominant Subhealth and Liver-Qi Stagnation and Spleen-Qi Deficiency Syndrome.

To observe the effects of Xiaopiyishen Herbal Extract Granule (XPYS-HEG) on the quality of life in people with fatigue-predominant subhealth (FPSH) and liver-qi stagnation and spleen-qi deficiency syndrome, the participants were allocated randomly to the treatment group (XPYS, n = 100) and the control group (placebo, n = 100) in this study. The study period was 18 weeks (6 weeks for the intervention and 12 weeks for followup). The results show that there were no differences between the two groups for the scores of eight factors on the SF-36 (Chinese version of the SF-36 universal quality-of-life scale) at baseline. Compared with the baseline score, intervention with XPYS-HEG led to a significant increase in scores for the factor of bodily pain at the end of the 6th week. Compared with the score at the end of the 6th week, the score for the factor of mental health in the XPYS group significantly increased at the end of the 18th week. Therefore, XPYS-HEG could partially improve the quality of life for people with FPSH and liver-qi stagnation and spleen-qi deficiency syndrome, which can ease bodily pain, stimulate a positive mood, and ease a negative mood.

[Weighting coefficients of symptoms and signs in the diagnosis of corresponding TCM syndrome elements of ulcerative colitis based on expert questionnaire investigation].

OBJECTIVE: To define the weighting coefficients of the symptoms and signs in the diagnosis of corresponding traditional Chinese medicine (TCM) syndrome elements of ulcerative colitis based on expert questionnaire investigation. METHODS: The questionnaire included eight syndrome elements of location and 11 syndrome elements relating to disease characteristics. Experts who answered the questionnaire were titled as associate chief physician or above, and had been practicing as professionals in general practice of medicine of digestive system for a long time. They came from 15 third-grade class-A hospitals (6 provinces and Beijing) in China. The weighting coefficients of the symptoms and signs in the diagnosis of corresponding TCM syndrome elements of ulcerative colitis were identified preliminarily based on consent degree of the experts (mean value and variation coefficient). RESULTS: There were 9, 6, 14, 5, 9, 8, 12 and 2 symptoms and signs corresponding to 8 TCM syndrome elements related to disease location and the ranges of weighting coefficients were listed as follows: liver, from 7.49 to 4.18; heart, from 6.90 to 5.51; spleen, from 7.96 to 5.55; lung, from 6.30 to 5.27; kidney, from 7.82 to 5.71; stomach, from 7.53 to 6.15; large intestine, from 8.40 to 6.70; bowel collaterals, from 8.49 to 6.41. Numbers of symptoms and signs corresponding to 11 TCM syndrome elements related to disease nature were 13, 8, 17, 18, 17, 18, 6, 11, 2, 10 and 11 and the ranges of weighting coefficients were listed as follows: qi deficiency, from 7.44 to 5.60; blood deficiency, from 7.90 to 5.59; yin deficiency, from 6.88 to 4.91; yang deficiency, from 7.54 to 5.57; dampness, from 7.91 to 4.96; excess heat (fire), from 7.13 to 5.69; excess cold, from 7.51 to 6.14; qi stagnation, from 7.38 to 5.88; qi sinking, from 7.65 to 7.43; blood stasis, from 7.60 to 5.79; fluid retention, from 7.08 to 4.46. CONCLUSION: On the basis of previous document research and clinical research in ulcerative colitis, the expert questionnaire can collect consensus opinions of experts. The weighting coefficients of the symptoms and signs in the diagnosis of corresponding TCM syndrome elements of ulcerative colitis are defined preliminarily. Thereby it will lay the foundation for establishing differentiation criteria of ulcerative colitis based on TCM syndrome elements.

[Effects of Xiaopi Yishen herbal extract granules in treatment of fatigue-predominant subhealth due to liver-qi stagnation and spleen-qi deficiency: a prospective, randomized, placebo-controlled and double-blind clinical trial].

BACKGROUND: The demand for effective intervention for subhealth conditions is growing with increasing numbers of people being in a state of subhealth with a poor quality of life. Future research and evaluation of the treatment methods for subhealth conditions from the perspective of traditional Chinese medicine (TCM) may provide an important direction for developing effective management of these conditions. OBJECTIVE: To evaluate the efficacy and safety of Xiaopi Yishen herbal extract granules (XPYS-HEG), a compound traditional Chinese herbal medicine for relieving fatigue and promoting a cheerful spirit for the treatment of people with fatigue-predominant subhealth due to liver-qi stagnation and spleen-qi deficiency. DESIGN, SETTING PARTICIPANTS AND INTERVENTIONS: A multi-center, randomized, double-blinded, placebo-controlled clinical study was undertaken. The study period was 18 weeks, including 6 weeks for intervention and 12 weeks for follow-up. Participants were recruited from medical center and outpatient clinics of three hospitals in China, i.e. Xiaotangshan Hospital of Beijing, the First Affiliated Hospital of Henan University of TCM and the Affiliated Hospital of Liaoning University of TCM. Two hundred participants who met the criteria of fatigue-predominant subhealth and liver-qi stagnation and spleen-qi deficiency in TCM were allocated randomly to the treatment group (XPYS, n=100) and control group (placebo, n=100). MAIN OUTCOME MEASURES: The total score of Fatigue Scale-14 (FS-14) was used to evaluate the fatigue status of subjects and the extent of liver-qi stagnation and spleen-qi deficiency syndrome was also recorded. RESULTS: Three cases in the XPYS group withdrew from the trial. There were 200 subjects who entered to full analysis set (FAS) analysis and 197 subjects fitted in the per-protocol set (PPS) analysis. (1) According to the score changes of FS-14, the effectiveness rates in the XPYS and placebo group were as follows: 14.0% vs 9.0% (FAS) and 14.4% vs 9.0% (PPS) for complete remission, 19.0% vs 15.0% (FAS) and 19.6% vs 15.0% (PPS) for obvious effects, 39.0% vs 26.0% (FAS) and 39.2% vs 26.0% (PPS) for effective, and 72.0% vs 50.0% (FAS) and 73.2% vs 50.0% (PPS) for complete efficacy. The efficacy of XPYS-HEG was superior to the placebo statistically (P<0.05). (2) According to the score changes of TCM syndrome, the effectiveness rates in the XPYS group and placebo group were as follows: 1.0% vs 0.0% (FAS) and 1.0% vs 0.0% (PPS) for complete remission, 20.0% vs 7.0% (FAS) and 19.6% vs 7.0% (PPS) for obvious effects, 29.0% vs 24.0% (FAS) and 29.9% vs 24.0% (PPS) for effective, and 50.0% vs 31.0% (FAS) and 50.5% vs 31.0% (PPS) for complete efficacy. The efficacy of XPYS-HEG was superior to that of placebo statistically (P<0.05). (3) The follow-up results at 12 weeks and 18 weeks showed that the efficacy of XPYS-HEG was superior to that of placebo statistically (P<0.05). (4) No adverse effects were found in the XPYS group. CONCLUSION: It can be concluded that XPYS-HEG is effective and safe for the treatment of people with fatigue-predominant subhealth due to liver-qi stagnation and spleen-qi deficiency.

[Common syndrome factors of traditional Chinese medicine in chronic renal failure based on the questionnaire investigation among experts].

OBJECTIVE: To screen common traditional Chinese medicine (TCM) syndrome factors of chronic renal failure (CRF) via questionnaire investigation among experts. METHODS: A questionnaire was developed based on the results of our previous researches. The investigation was carried out with the questionnaire among experts who are engaged in clinical research and treatment of nephrosis with the title of chief or associate chief physician from 15 hospitals in China. Common TCM syndrome factors of CRF were obtained based on the data analysis of the mean value on general evaluation, cumulative percentage and coefficient of variation. RESULTS: A total of 114 sets of the questionnaire were sent out and all of them were returned back, 113 of which were effective. The recovery rate is 100%. The TCM syndrome factors were regarded as common factors of CRF syndrome if the cumulative percentage was more than or equal to 60% and the coefficient of variation was less than or equal to 0.35. The syndrome factors related to the disease location were the kidney, spleen, stomach, heart, and lung; the syndrome factors related to the disease type were dampness, turbid toxin, blood stasis, water retention, and phlegm which belong to excess type, and qi deficiency, yin deficiency, yang deficiency, blood deficiency, and essence deficiency which belong to deficiency type. CONCLUSION: The common TCM syndrome factors of CRF were obtained from the representative experts through the questionnaire investigation among the experts. The results enable us to have a deeper understanding of the disease in view of TCM compared with the previous literature data and may contribute to the establishment of TCM syndrome diagnosis criteria of the disease in the future.

[Distribution characteristics of traditional Chinese medicine syndromes and their elements in people with subhealth fatigue].

OBJECTIVE: To investigate the characteristics of traditional Chinese medicine (TCM) syndromes and their elements in people with subhealth fatigue. METHODS: The TCM symptoms in line with the diagnostic criteria of subhealth fatigue status were collected by clinical investigations and using information collection form based on TCM four diagnostic methods. Referred to Clinical Guidelines of Chinese Medicine on Subhealth and other related standards, the syndrome type was identified in accordance with clinical symptoms of each patient with subhealth fatigue by two physicians. The data of syndrome differentiation were analyzed by descriptive statistical analysis. RESULTS: There were 81 syndrome types from 495 cases of subhealth fatigue. There were 24 syndrome types after separation, and the top ten were liver stagnation and spleen deficiency, stagnation of liver qi, hyperactivity of liver fire, disharmony between liver and stomach, damp obstruction due to spleen deficiency, deficiency of both heart and spleen, yin deficiency of liver and kidney, yang deficiency of spleen and kidney, stagnation of gallbladder and disturbance of phlegm, and internal disturbance of phlegm-heat. There were 17 syndrome elements, including seven disease location elements and ten disease nature elements. The disease location elements were liver, spleen, kidney, stomach, heart, gallbladder and lung. The disease nature elements were qi stagnation, qi deficiency, exuberance of fire (heat), damp obstruction, phlegm obstruction, yin deficiency, adverse flow of qi, yang deficiency, blood deficiency, and blood stasis. CONCLUSION: Syndrome types of subhealth fatigue involve in deficiency syndrome, excess syndrome, and mixture of deficiency and excess syndromes. The syndrome elements of disease location involve five zang organs and two fu organs, and the liver and spleen were the most frequently involved organs. The syndrome elements of disease nature involve deficiency and excess. Qi stagnation is most frequently involved in the excess syndrome, and qi deficiency is most frequently involved in the deficiency syndrome.

[A perspective of developing syndrome diagnostic criteria based on syndrome factors and combination of disease in modern medicine system and syndrome in traditional Chinese medicine system].

Syndrome differentiation treatment is the traditional model of diagnosis and treatment of diseases in traditional Chinese medicine (TCM). To establish scientific diagnostic criteria of TCM syndrome is one of the key points in TCM study. In this paper, the basic models of the relevant diagnostic criteria of TCM syndrome and existed problems were reviewed. The authors pointed out the advantages of establishing diagnostic criteria of TCM syndrome based on TCM syndrome factors and combination of disease in Western medicine system and TCM syndrome, in which not only the characteristics of the disease in Western medicine were considered, but also the complexity and flexibility of syndrome identification and convenient application in clinical practice were resolved. The basic model and frame of the above diagnostic criteria and the procedures and methods used in developing the diagnostic criteria were also described and discussed.

HERG-F463L potassium channels linked to long QT syndrome reduce I(Kr) current by a trafficking-deficient mechanism.

1. Congenital long QT syndrome (LQTS) is a genetically heterogeneous disease. The aim of the present study was to identify the gene mutation in a Chinese family with LQTS and investigate the functional changes associated with the mutation. 2. Polymerase chain reaction and DNA sequencing were used to screen for the KCNH2 mutation in the proband. A mutant F463L HERG channel was expressed in HEK293 cells using a lipofectamine method. The IKr current was recorded using the whole-cell voltage clamp technique. Expression of HERG protein was detected by western blotting and the subcellular location of HERG channels in cell was analysed by confocal microscopy. 3. The novel heterozygous missense mutation F463L in KCNH2 was detected. We found that the F463L mutation did not lead to any expression of detectable I(Kr) current, which was consistent with western blotting analysis indicating that the F463L mutation only expressed a band at 135 kDa. When coexpressed with wild-type HERG, F463L HERG exhibited strong dominant-negative current suppression, resulting in a decrease in I(Kr) current density, and induced a positive shift in the voltage dependence of activation, as well as interference with trafficking of wild-type channel protein. The processing of the F463L channels was partly corrected in cells incubated in E4031. In addition, confocal microscopy demonstrated that F463L subunits could be inserted into the cell membrane when forming heteromultimeric channels with wild-type channel subunits. 4. The results of the present study suggest that the F463L mutation leads to loss of function in HERG through a dominant-negative effect caused by impaired trafficking of the channel.

[Characteristics of traditional Chinese medicine syndromes and their element distributions in sub-health status: a modern literature review].

OBJECTIVE: To probe into the characteristics of traditional Chinese medicine syndromes and their element distributions in sub-health status. METHODS: Literatures on sub-health in past 20 years were collected, and a data bank was set up by using EpiData 2.0. The frequencies of syndromes and their elements were analyzed using SPSS 12.0. RESULTS: Out of the 50 syndromes obtained from the standardization of syndrome nomenclature, and the top three were stagnation of liver-qi, deficiency of both heart and spleen, and deficiency of liver-yin and kidney-yin. Spleen, liver and kidney were the top three of all the 14 disease locations. Qi-deficiency, qi-stagnation and damp pathogen were the top three pathogenicity types. CONCLUSION: Although the traditional Chinese medicine syndromes in sub-health status are scattered in the distribution, the main syndromes take up a leading proportion and their elements are concise and concentrated in distribution. Therefore, it is necessary to make a standard study on traditional Chinese medicine syndromes in sub-health status from syndrome elements.

[Estimation on the reliability and validity of the fatigue self-assessment scale].

OBJECTIVE: To analyze the reliability and validity of the Fatigue Self-assessment Scale (FSAS). METHODS: The scale was applied among the participants assigned to 4 groups, the differences in types, degrees and characteristics of fatigue of them were compared, and the reliability and constitutional validity of ESAS were assessed by internal consistency analysis, exploratory factor analysis and confirmatory factor analysis using the statistical software of SPSS and LISREL. RESULTS: Statistical differences of types, degrees and characteristics of fatigue presented in the participants of the 4 groups. The Cronbach's alpha of various factors in the scale were 0.772-0.908; the indexes for the section of assessing type, and degree of fatigue were RMSEA=0.065, NNFI=0.95, CFI=0.96; and those for the section of assessing characteristics of fatigue were: RMSEA=0.10, NNFI=0.93, CFI=0.96. CONCLUSION: The FSAS has good differentiability, reliability and constitutional validity for assessing the type, degree and characteristics of fatigue in various populations. In order to explore the relationship of TCM syndrome patterns with the type, degree and characteristics of fatigue, its future application for evaluation of fatigue and intervention effect of anti-fatigue should be combined with TCM syndrome differentiation.

[Sub-health state and chronic fatigue syndrome].

This paper points out that the sub-health state is not equal to chronic fatigue syndrome (CFS) on basis of elaborating the concept and category of sub-health. And the present understanding on concepts of fatigue, chronic fatigue and CFS, as well as the diagnosis criteria and differential diagnosis of CFS are discussed systematically.

[Functional expression of congenital long QT syndrome related HERG mutation A561V in vitro].

OBJECTIVE: To investigate the functional expression of HERG mutation A561V detected in a Chinese congenital long QT syndrome family. METHODS: The mutation gene A561V was cloned into eukaryotic expressive vector pcDNA3 by quick site-directed mutagenesis PCR and restriction enzymes. The wild-type HERG, heterozygous type HERG and HERG mutation A561V were respectively cotransfected with pRK5-GFP into HEK293 cells by Suprefact transfection regent. The protein expression was measured by immunofluorescence method and Western blot. The electrophysiological characteristics of transfected cells were determined by whole cell patch-clamp technique. RESULTS: Direct sequence analyses revealed a C to T transition at position 1682. A561V mutation was correctly combined to eukaryotic expressive vector pcDNA3 and expressed in HEK293 cells. The protein expression of mutation and heterozygosis were located in cytoplasm and cellular membrane. 155 kDa and 135 kDa protein bands were detected in wild type HERG channel while only 135 kDa protein band was shown in heterozygous and mutational channels. Significant HERG tail-current was recorded in wild type HERG channel but not in mutation and heterozygosis channels. CONCLUSION: This study evidenced a functional dominant-negative current suppression in HEK293 cells transfected with HERG mutation A561V.

[Construction of eukaryotic expression vector of congenital long QT syndrome related HERG gene A561V mutation and its expression in HEK293 cells].

OBJECTIVE: To investigate the protocol of the construction of HERG gene mutations, an A561V mutation which was detected in a Chinese congenital long QT syndrome (LQTS) family had been constructed and expressed in vitro. METHODS: The A561V cloning vector PGEM-HERG-A561V was constructed by quick site-directed mutagenesis PCR. The A561V expressive vector pcDNA3-HERG-A561V was constructed by restriction enzymes. pRK5-GFP was cotransfected with pcDNA3-HERG-A561V or wild type pcDNA3-HERG into HEK293 cells by Superfect transfection reagent. The protein was measured by immunofluorescence. RESULTS: Direct sequence analyses revealed a C to T transition at position 1682. The A561V mutation was correctly combined to eukaryotic expressive vector pcDNA3 and expressed in HEK293 cells. The protein of mutation was expressed in cytoplasm and cellular membrane while the wild type gene was expressed only on cellular membrane. CONCLUSION: The protocol can be used successfully to construct and express HERG A561V mutation and it forms the basement of the further study on functions of mutation.

[Long QT syndrome gene diagnosis by haplotype analysis].

OBJECTIVE: Three long QT syndrome(LQTS) pedigrees were brought together for genetic diagnosis by using short tandem repeat(STR) markers. METHODS: Genomic DNA was extracted from blood samples. STR markers (D7S1824, D7S2439, D7S483, D3S1298, D3S1767, D3S3521) in or spanning the HERG and SCN5A gene were amplified; the haplotype analysis for LQTS was performed. RESULTS: Clinical diagnosis showed that 15 are LQTS patients (3 died) and 11 are probable patients. Linkage analysis showed that LQTS patients are linked with the SCN5A gene in family 1, HERG is linked with the disease in family 2 and 3. Fourteen gene carriers were identified, 2 patients and 7 probable patients were excluded. CONCLUSION: Linkage analysis using STR markers can serve as useful tool for presymptomatic diagnosis.

[Preparation of polyacrylamide gel electrophoresis for human chorionic gonadotropin chimeric peptide 12 expressed in E. coli].

In recent years, development of chimeric peptide (CP) immunogens is a trend in the vaccinological field. The CPs contain a B cell epitope(s) of target antigen and a promiscuous self - or foreign- T cell epitope(s). However, such constructed CPs were all expressed in prokaryotic or eukaryotic systems at lower levels. To purify the human chorionic gonadotropin (hCG) CP12 expressed in E. coli at the level of about 1% of the total cell proteins, an improved method of preparative gel polyacrylamide gel electrophoresis (PAGE) was developed. The important improvement to routine preparative PAGE involves: (1) running reversed electrophoresis by rearranging the gel- carrying plate when the bromophenol blue band arrived at 1-1.5 centimeter from the bottom of the gel; (2) making a collecting trough between the gel and a dialytic membrane that was used to isolate the upper tank buffer. About 8 fractions were collected at regular intervals of 15 minutes after bromophenol blue running out of gel. And then 0.2 ml was taken from each fraction and the protein was precipitated by sequentially adding trichloroacetic acid and acetone. Each sample was dissolved in 20 microL sample buffer and analyzed and identified by SDS-PAGE and Western blotting. As a result, the hCG CP12 expression product with 95% relative homogeneity was harvested at a 50-100 microgram level after a single-step purification of this preparative PAGE, with respect to the sample which contained 3-4 mg of cell proteins.

[Double action of dopamine on PC12 cells].

PC12 cell line was used to examine the cytotoxicity of dopamine (DA). The higher dose of DA (400micromol/L) greatly reduced cell survival rate and induced apoptosis,meanwhile the lower dose (100micromol/L) had less cytotoxicity. Further studies showed that AMP (100micromol/L) were not cytotoxic at 24h, but decreased cell survival rae to 78% at 72h compared to control, and also exacerbate the cytotoxicity of DA (100micromol/L) at 72h. By determine the action of GSH level and SOD activity, we found that appropriate intracellular DA may be advantageous to protect cells from the toxicity of higher dose DA. The intracellular calcium concentration showed that DA's toxicity increased intracellular concentration of calcium. The results suggest that it was the extracellular DA that made cytotoxicity by decreasing cell's possibility of antioxide and increasing intracellular free calcium.