Objective To investigate DNA methylation in the promoter region,mRNA transcription and protein expression of glutathione-S-transferases-P1 (GSTP1) gene and their relation with arsenism.Methods In endemic coal-pollution-borne arsenism area,Jiaole village of Xinren county,Guizhou province,according to the diagnostic criteria of endemic arsenism(WS/T 211-2001),123 cases with endemic arsenism were selected and divided into three groups (mild arsenism group:42 cases,moderate arsenism group:41 cases and severe arsenism group:40 cases).Forty seven residents were selected as controls in a village about 12 km away from the endemic arsenism area.With the informed consent principle,peripheral blood of all respondents was collected in order to analyze DNA methylation and check mRNA.DNA methylation of GSTP1 gene promoter region in peripheral blood was assayed by PCR,and GSTP1 mRNA expression was assayed using real-time quantitative PCR.In addition,other cutaneous specimens originated from 53 cases with arsenism that accepted surgical treatment voluntarily were taken.Of these specimens,general pathological changes were 28 cases,precancerous 20 cases and cancerous 5 cases.Skin tissues of 15 cases of non-tumor surgery patients without abnormal pathological changes were as control group.GSTP1 protein expression in the skin tissue was detected using immunohistochemistry (IHC).Results Among different groups of arsenic poisoning,the positive rate of DNA methylation of GSTP1 gene was 28.57％(12/42) in the mild group,57.10％ (23/41) in the moderate group and 65.00％ (26/40) in the severe group.Compared with the control group (6.38％,3/47),the difference was statistically significant (x2 =7.792,26.000,33.412,all P ＜ 0.01).Among different groups of arsenic poisoning diagnosed by dermapathology,the positive rate of DNA methylation of GSTP1 gene was 21.43％(6/28) in the general pathological change group,50.00％(10/20) in the precancerous group and 80.00％(4/5) in the cancerous group.Compared with the control group(6.67％,1/15),the difference was statistically significant (x2 =3.562,7.468,10.756,all P ＜ 0.05).It showed that the positive rate of DNA methylation of GSTP1 gene increased with aggravation of the disease and dermatic lesion of arsenism (tendency x2 =38.239,x2 =13.659,all P ＜ 0.01).Compared with the control group(0.184 26),the expressions of GSTP1 mRNA in peripheral blood in moderate (0.087 77) and severe arsenic poisoning groups (0.056 93) were significantly reduced(all P ＜0.01),and that of severe group was significantly lower than that of the moderate group (P ＜ 0.01) ; compared with the control group(0.338 45) and the general lesion group(0.276 74),GSTP1 mRNA expression was significantly reduced in precancerous lesion group(0.104 81) and cancerous group(0.043 70),in which the cancerous group was significantly lower than that of the precancerous lesions.The difference of skin tissue GSTP1 protein expression rate between groups was statistically significant (x2 =20.948,P ＜ 0.05),in which the difference between the precancerous lesion group(65.00％,13/20),the cancer group (40.00％,2/5) and the control group(100.00％,15/15)was statistically significant (x2 =12.183,11.778,P ＜ 0.01).Spearman correlation analysis showed that the degree of skin lesion and the level of GSTP1 protein expression was negatively correlated (r =-0.520,P ＜ 0.05).Groups were divided according to DNA methylation of GSTP1 gene,and the mRNA and protein expression of GSTP1 in methylation group(0.038 40,57.14％) was significantly lower compared with that of unmethylated group(0.187 07,95.74％; Z =9.032,x2 =23.134,all P ＜ 0.01).Conclusions Arsenism may lead to DNA methylation of human GSTP1 gene promoter region,thereby inhibiting expression of mRNA and protein.GSTP1 gene plays an important role in arsenism or carcinogenic process.

Epithelial-Mesenchymal Transition ; Humans ; Liver Cirrhosis

Humans ; Liver Cirrhosis ; classification ; pathology ; Pathology, Clinical ; Terminology as Topic

OBJECTIVETo evaluate the genotoxicity of a magnesium alloy coated with beta-tricalcium phosphate (beta-TCP).

METHODSFour groups were designed. In the first group, AZ31B magnesium alloy surface was coated with beta-TCP using chemical bath deposition, and in the second group magnesium alloy was tested. The other two groups were negative control (pure titanium) and positive control groups (0.5 mg/L bleomycin). Single cell gel electrophoresis was adopted to investigate genotoxicity of the alloy samples in different groups, and 60 cells from each group were analysed. Tail moment and tail DNA percentage were used as reliable indicators to show DNA damage in lymphocytes induced by every testing sample. Student-Newman-Keuls (SNK) test was used to compare results from 4 groups.

RESULTSThere were no significant differences in tail moment and tail DNA percentage between magnesium alloy group [(0.52 +/- 0.12), (6.82 +/- 1.81)%] and magnesium alloy coated with beta-TCP group [(0.51 +/- 0.12), (6.89 +/- 1.93)%, P > 0.05]. Tail moment and tail DNA percentage in negative group were (0.47 +/- 0.14) and (6.29 +/- 1.64)%, and tail moment and tail DNA percentage in positive group were (5.17 +/- 1.23) and (22.09 +/- 4.51)%.

CONCLUSIONSNo significant increase was found in DNA damage in lymphocytes induced by magnesium alloy coated with beta-TCP.

Alloys ; Calcium Phosphates ; Coated Materials, Biocompatible ; Electrophoresis, Agar Gel ; Humans ; Magnesium ; Materials Testing

A rapid and sensitive analytical method was developed for the simultaneous determination of fumonisins B1 and B2 in traditional Chinese medicines by HPLC-MS/MS. The detection limits for fumonisins B1 and B2 were 0.25 ng x mL(-1) corresponding to 2 microg x kg(-1) in samples. Recoveries from different samples spiked with fumonisins B1 and B2 at levels ranging from 0.2 to 3 mg x kg(-1) were 84.0%-96.1% and 86.3%-99.3%, respectively. Among the total of 34 samples purchased from local markets, ten samples of which were visibly moldy samples due to inappropriate storage, and 24 were normal samples. The results showed that 6 of the visibly moldy samples and 5 of the normal samples were contaminated with total fumonisins at levels ranging 82.4-2349 microg x kg(-1) and 102-729 microg x kg(-1), respectively.
Chromatography, High Pressure Liquid ; Drug Contamination ; Drugs, Chinese Herbal ; chemistry ; Fumonisins ; analysis ; Medicine, Chinese Traditional ; Plants, Medicinal ; chemistry ; Tandem Mass Spectrometry

OBJECTIVETo establish the criteria of the disproportionate loss of Mandarin monosyllable discriminative abilities to pure tone hearing thresholds.

METHODSTotal of 165 patients with varying degrees of sensorineural hearing loss were recruited for routine audiological evaluations. The speech discrimination scores were obtained by Mandarin phonemic-balanced monosyllable lists via self-made speech audiometric software. The Performance-Intensity (P-I) function for individual ear was obtained by the same list which was administrated in ascending intensities, with 25 monosyllables presenting randomly. The lowest intensity was determined by the lowest pure tone threshold among all audiometric frequencies minus 5 dB. The intensities were increased in 5 dB step until the score was 100% or the intensity was reached to the patient's uncomfortable level. The PB(max) was obtained from the P-I plot. Three parameters about pure tone average hearing thresholds, including PTA(1) (average of 0.5, 1 and 2 kHz), PTA(2) (average of 1, 2 and 4 kHz) and PTA(3) (average of 0.5, 1, 2 and 4 kHz), as well as three parameters about audiogram slope, including Slope(0.5) (4 kHz minus 0.5 kHz), Slope(1) (4 kHz minus 1 kHz) and Slope(2) (4 kHz minus 2 kHz), were calculated respectively. The correlations between PB(max) and above parameters were analyzed by SPSS10.0 statistical software.

RESULTSThe audiogram slopes were not shown any correlation with PB(max), while the pure tone average thresholds, especially PTA(3) (r = -0.595, P = 0.000) were confirmed to correlate with PB(max). In the scatter plot based on PB(max) and PTA(3), a linear boundary was constructed encompassing approximately 99% of observed data collected from the sensorineural hearing-impaired.

CONCLUSIONAny PB(max) score falling below the boundary should be considered with high possibility and disproportionately poor comparison with pure tone hearing thresholds.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Audiometry, Pure-Tone ; Audiometry, Speech ; Auditory Threshold ; Female ; Hearing Loss, Sensorineural ; physiopathology ; Humans ; Male ; Middle Aged ; Young Adult

Objective To comparatively analyze the changes of environmental risk factors in 9 years in an area polluted by arsenic coal-burning in Xingren County of Guizhou Province,in order to provide evidence for reasoning the occurrence and development as well as its effective prevention and control.Methods Epidemiological sampling methods was used to conduct follow-up investigation on 181 arsenism patients who were diagnosed in 1998 in arsenic polluted area.Control group included 65 residents who lived far from polluted area of 12 km.The follow up investigation included age,sexuality,family economic situation,time of use or stop use of arsenic coal, ventilation of the room,desiccation of food etc.Diethyl dithiocarbamate silver(Ag-DDC)method was used to detect arsenic content of coal,soil,air,water and rice,corn,chili;Single factor and multivariate factors non-conditional Logistic regression models were used to analyze exposure factors of patients and related environmental risk factom, and the differences of those in 1998 and 2006 were compared. Results The arsenic content in indoor and outdoor air,coal,chili and corn went down from 0.0880 and 0.0220 mg/m3,397.20,45.07 and 2.64 mg/kg in 1998 to 0.0790 and 0.0070 mg/m3,93.01,3.46 and 1.50 mg/kg in 2006. Arsenic contents of other samples were less than national standard. The analysis of single factor and multivariate factors non-conditional Logistic regression showed that time of using high arsenic coal,age,fluorosis and smoking(x2 = 50.159,12.195,37.69,6.358,P ＜ 0.05 or ＜ 0.01 ) were still the main risk factors for arsenism,while family economic situation was still influential factors (X2 = 4.614,P ＜ 0.05);Ventilation of the room changed from a risk factors at 1998 to an influencing factors at 2006(X2 = 38.093,P ＜ 0.01 ). Single factor non-conditional Logistic regression model analysis showed that food desiccation by arsenic coal-burning and educational level were no longer risk and influencing factors,while food preservation and gender had become influencing factors(x2 = 17.463,11.004,all P ＜ 0.01 ) nine years after. Conclusions Environmental arsenic pollution in arsenism area in Guizhou Province has been obviously improved after nine years. However,the continued existence of low doses of arsenic pollution is still a major cause of failure of controlling arsenism. Time of using high arsenic coal,age,smoking,fluorosis,family economic situation and ventilating room are closely related to the occurrence and the development of arsenism. Prohibition of use of high arsenic coal,furnace improvement,health education and economic development are effective measures

OBJECTIVETo explore a treatment program for increasing therapeutic effect on rheumatoid arthritis at active stage.

METHODSOne hundred and forty-six cases were randomly divided into treatment group (n = 74) and medicine control group (n = 72). The treatment group were treated by electroacupuncture at Quchi (LI 11), Hegu (LI 4), Yanglingquan (GB 34), etc. , combined with meloxicam, sulfasalazine and MTX. The control group treated by simple the Western medicines. Their therapeutic effects were compared.

RESULTSThe effective rate was 79.73% in the treatment group and 51.39% in the control group with a significant difference between the two groups (P< 0. 05).

CONCLUSIONElectroacupuncture combined with medicine has a better therapeutic effect than the simple medicine on rheumatoid arthritis at active stage.

Arthritis, Rheumatoid ; therapy ; Electroacupuncture ; Humans ; Medicine

Markov model is one of the decision analysis models,which is widely applied in pharmacoeconomic evaluation.The acquisition of transition probability parameters among different Markov health states were difficult in model building and hard to be acquired directly.It discussed the calculation of current Markov model transition probability parameters,especially for the time-dependent model.Furthermore,it performed a case analysis to explore how to calculate the transition probability and provide references for researchers of related research.

OBJECTIVETo establish an efficient genetic transformation system of Pinellia ternata.

METHODWith petioles from test-tube seedlings of P. ternata as explants, Agrobacterium tumefaciens mediation method was adopted to explore the effect of phenolic substances, A. tumefaciens's concentration, infection time, pre-incubation time and co-cultivation time on genetic transformation efficiency of P. ternata.

RESULT AND CONCLUSIONThe genetic transformation efficiency could be effectively enhanced by infecting in A. tumefaciens culture containing AS 40 mg x L(-1) for 15 min for three days. The petioles were put into the differentiation medium containing 150 mg x L(-1) Kan and 350 mg x L(-1) Carb to screening and cultivation. After around 30 days, microtubers could be observed at both sides of the petioles. Gus staining and PCR verification on the regenerated plants showed that the exogenous gene sHSP had been integrated into genome of P. ternata.

Agrobacterium tumefaciens ; genetics ; DNA, Plant ; genetics ; Genetic Engineering ; methods ; Glucuronidase ; genetics ; metabolism ; Heat-Shock Proteins, Small ; genetics ; Pinellia ; genetics ; growth & development ; metabolism ; Plant Leaves ; genetics ; growth & development ; metabolism ; Plants, Genetically Modified ; Polymerase Chain Reaction ; Reproducibility of Results ; Tissue Culture Techniques ; methods ; Transformation, Genetic