Objective To study the expression of double-stranded RNA-dependent protein kinase (PKR) in malignant melanoma and ordinary nevi. Methods The expression of PKR and proliferating cell nuclear antigen was examined in 42 cases of malignant melanoma and 25 ordinary nevi by an immunohistochemical method. Results The positive rate of PKR expression was higher in the patients with malignant melanoma than that in the patients with ordinary nevi (P

Objective To estimate the relationship of the functional single nucleotide polymorphism (SNP) rs1052133 in the 8-oxoguanine DNA glycosylase 1 (OGG1) gene with vitiligo in a Chinese Han population.Methods Blood samples were collected from 800 patients with vitiligo and 800 healthy human controls,and subjected to genomic DNA extraction.PCR-restriction fragment length polymorphism (PCR-RFLP) analysis was performed to analyze the genotype of the SNP rs1052133 in the OGG1 gene.The relationship between the SNP and the risk of vitiligo was evaluated by chi-square test and unconditional logistic regression analysis.Enzyme linked immunosorbent assay (ELISA) was carried out to assess the serum level of 8-hydroxydeoxyguanosine (8-OHdG) in 83 patients with vitiligo and 83 healthy human controls,then,t test was used to compare the serum 8-OHdG level between the patients and controls.Results The frequency of CC,CG and GG genotype of the SNP rs1052133 was 16.8％,54.0％ and 29.2％ respectively in the patients,21.4％,52.8％ and 25.8％respectively in the controls (x2 =6.26,P ＜ 0.05).Increased frequency of G allele of the SNP rs1052133 was observed in the patients with vitiligo compared with the controls (56.2％ vs.52.2％,x2 =5.16,P ＜ 0.05).A statistically increased risk of vitiligo was associated with the CG (x2 =3.98,P ＜ 0.05,adjusted odds ratio 1.31,95％ confidence interval:1.01-1.70) and GG (x2 =6.01,P ＜ 0.05,adjusted odds ratio 1.45,95％ confidence interval:1.08-1.94) genotype of SNP rs1052133 compared with the CC genotype,which was more evident among the patients with the following characteristics:female,nonsegmental vitiligo,active vitiligo,long clinical course (＞ 12 months),a family history of vitiligo,and no accompanied autoimmune diseases.In addition,the patients with the CG or GG genotype of SNP rs1052133 had a higher serum 8-OHdG level than those with the CC genotype ((838.23 ± 294.11) μg/L vs.(593.84 ± 190.14) μg/L,t =3.63,P ＜ 0.01).Conclusions The SNP rs1052133 in the OGG1 gene may be responsible for the development of vitiligo in Chinese Han populations,which is likely to be associated with defects in DNA repair.

Objective To investigate human papilloma virus(HPV) infection in normal endometrium, atypi-cal hyperplasia of endometrium and endometrial carcinoma. Methods By the nucleic acid hybridization,we detected 28 pairs of endometrial carcinomas, 21 pairs of atypical hyperplasia of endometrium. Normal endometrium from 16 pa-tients with uterine myomas were as control. Results HPV16/18 DNA was detected in 25 endometrial carcinoma and 2 atypical hyperplasia of endometrium and 1 normal endometrium. Conclusions Endometrial carcinoma HPV16/18 DNA were significantly higher than those infected with atypical endometrial hyperplasia and normal endometrium. Note the occurrence of endometrial cancer,the development may be associated with HPV infection.

Objective To investigate the clinical effects of lactobacillus acidophilus combined with glutathione in the treatment of nonalcoholic fatty liver disease.Methods 120 patients with nonalcoholic fatty liver disease were selected,and they were randomly divided into control group (60 patients used glutathione alone) and observation group (60 patients received lactobacillus acidophilus on the basis of control group).The clinical efficacy,liver function index,blood lipid index and the ultrasonography grading of fatty liver before and after treatment of the two groups were compared.Results The clinical total effective rates of the control group and the observation group were 76.67％,91.67％,respectively.The total effective rate of the observation group was significantly higher than that of the control group(x2 =10.52,P ＜ 0.05).The levels of AST,ALT and GGT of the observation group after treatment were significantly lower than those of the control group and before treatment [(36.89 ± 7.14) U/L,(36.89 ± 7.14) U/L,(36.89 ±7.14) U/L vs.(49.36 ± 11.08) U/L,(45.91 ± 10.24) U/L,(90.28 ± 20.70) U/L;(36.89 ± 7.14) U/L,(36.89 ± 7.14) U/L,(36.89 ± 7.14) U/L vs.(92.90 ± 24.3) U/L,(81.11 ± 17.37) U/L,(147.27 ± 34.19) U/L,t =2.88,2.54,2.91;3.01,3.36;3.18,3.48;3.41,3.87;all P ＜0.05).The levels of TG,TC,HDL-C and LDL-C of the observation group after treatment were significantly better than those of the control group and before treatment [(1.66 ± 0.42) mmol/L,(4.05 ± 0.77) mmol/L,(1.69 ± 0.60) mmol/L,(2.44 ± 0.38) mmol/L vs.(2.13 ± 0.80) mmol/L,(4.64 ± 0.94) mmol/L,(1.45 ± 0.48) mmol/L,(2.97 ± 0.57) mmol/L;(1.66 ± 0.42) mmol/L,(4.05 ± 0.77) mmol/L,(1.69 ± 0.60) mmol/L,(2.44 ± 0.38) mmol/L vs.(2.71 ± 1.33) mmol/L,(5.42 ± 1.27) mmol/L,(1.08 ± 0.36) mmol/L,(3.49 ± 0.71) mmoL/L,t =2.43.2.13,2.55,2.07;3.02,3.41;3.11,3.65;2.81,3.30;2.87,3.15;all P ＜ 0.05).In the control group,normal in 6 cases,mild in 24 cases,moderate in 21 cases and severe in 9 cases as fatty liver degree;in the observation group,normal in 13 cases,mild in 30 cases,moderate in 13 cases and severe in 4 cases as fatty liver degree.The ultrasonography grading of fatty liver of the observation group after treatment were significantly better than those of control group and before treatment (U =3.74,5.20,all P ＜ 0.05).Conclusion Lactobacillus acidophilus combined with glutathione in the treatment of nonalcoholic fatty liver disease can efficiently promote the recovery process of liver function,regulate blood lipid levels and is helpful to improve the imaging grading.

Objective: To study the inhibition of antisense TRP1 on cell growth of malignant melanoma(MM) and explore a new way for therapy of melanoma. Methods: Antisense TRP-1 recombinant vector was constructed and transfected into MM cells. According to the results of MTT, cell growth curves were drawn and then clonogenic assay was performed in vitro. At last, tumorigenesis assay was undertaken in nude mice in vivo. Results: Cell proliferations of TRP-1 transfected MM cells were inhibited compared with the control cells. The results of clonogenic assay displayed the difference of clonogenic percentage between TRP-1 transfected MM cells (52% , P

Objective To investigate melanocyte membrane antigen associated with vitiligo,for further purification and cloning.Methods Sera from patients with vitiligo were screened by using a live cell enzyme-linked immunoabsorbent assay,98strongly positive sera were obtrained.Melanocyte antigens were immunoblotted after splitting of cultured normal human melanocytes.Thirty strong positive vitiligo sera were detected.Results There were positive bands in all screened sera as shown by immunoblotting.Whereas,positive band was seen in only one normal human sera.The antibody bound many antigens with different molecular weights(150kd,90kd,75kd,50kd,40~45kd).Positive rates for individual antigens were70%,60%,83%,16%and23%,respectively.Conclusions Antibody directed to melanocyte membrane antigens of melanocytes are present in the sera of patients with vitiligo.The molecular weights of the antigens are mainly150kd,90kd and75kd,the antigens with small molecules discovered previously maybe the degradation products of big molecules.

Purpose:To study the value of nuclear DNA morphological parameters in differential diagnosis. Methods:Using the cellular image analysis instrument to quantitate nuclear DNA morphological parameters (including area, perimeter, major diameter, equivalent circle diameter and nuclear shape factor) in 38 cases of ovarian mucinous cystadenomatous and 5 normal ovarian epithelial tissue, and to analyze the relationship between histodifferentiation and nuclear morphological. Results:Statistical analysis showed that there were significant differences between the groups ( P 0.05). Conclusions:Nuclear DNA morphological parameters of ovarian mucinous cystadenomatous are useful markers, and may have some significance in pathological diagnosis.

Objective: To establish the method for determination of methyl acrylate, methybenzene, o dimethylb, m dimethy, p dimethy and phenylethene in Kechuanning Capsules(Papaver nudicaule) by chromatography. Methods: The GC system consisted of stainless steel column, 3.5% organic bentonite and 2.5% DNP as the solid phase, nitrogen as the carrier gas, and FID as the detector. Results: The contents of all the determined substances in Kechuanning Capsules are lower than 20mg?g -1 . Conclusion: The method is sensitive, accurate and reproducible, and it can be used to control the quality of the preparation.

Objective To construct the antisense eukaryotic vector of human TRP-1 (tyrosinase related protein 1) encoding gene, and transfect it into TRP-1 highly expressed melanocytes and malignant melanoma cell line, in order to further study the effects of antisense TRP-1 on the proliferation and functions of those cells. Methods TRP-1 cDNA was amplified by polymerase chain reaction (PCR) and the PCR products were subcloned into eukaryotic expression vector pcDNA3.1 on the opposite direction. Antisense recombinant vector was transfected into melanocytes and melanoma cell line. TRP-1 mRNA level was detected by reverse transcriptase polymerase chain reaction (RT-PCR). TRP-1 protein level was detected by Western blot. Cell cycle was determined by flow cytometry. The activity of tyrosinase was valued by L-dopa reaction. Results The recombinant antisense vector pcDNA3.1/TRP-1(-) was constructed. Positive transfected cells could steadily express TRP-1 antisense RNA. It was showed that there was a low level of TRP-1 mRNA as indicated by RT-PCR, and a low level of TRP-1 protein as indicated by Western blot. Cell cycles were blocked in G1 stage. The suppress rates of tyrosinase was 46% in transfected melanocytes and 54% in malignant melanoma cells, respectively. Conclusions TRP-1 plays an important role in the proliferation and functions of melanocytes and melanoma cells. Antisense TRP-1 could block the cell cycles and decrease the activity of tyrosinese in those cells.

Objective To investigate DJ-1 expression and protective effect against H2O2-induced oxidative stress in primary human melanocytes.Methods The expression and location of DJ-1 in primary melanocytes were identified by immunofluorescence.After cultured melanocytes were exposed to H2O2 for 24 hours,modified MTT assay was used to assess the proliferation of cells and to choose the suitable concentration of H2O2 for the following experiment.Western blot was used to detect D J-1 expression in melanocytes after being treated with 0.5 mmol/L H2O2 for 24 hours.Some melanocytes were divided into 3 groups to be reversely transfected with PBS(mock control group),non-targeting siRNA(negative control group)and DJ-1 targeting siRNA(DJ-1 group).Optical microscopy was utilized to observe the morphologic changes of transfected melanocytes.At 48 hours after the transfection,the melanocytes were stimulated with H2O2 for 24 hours.Subsequently,modified MTT assay,2′,7′-dichlorofluorescein diacetate(DCFH-DA)and annexin Ⅴ-fluorescein isothiocyanate/propidium iodide were used to determine cell viability,intracellular reactive oxygen species (ROS)level and apoptosis rate respectively.Results DJ-1 was expressed in both melanocyte nucleus and cytoplasm,and predominantly in the nucleus.H2O2 inhibited the cell viability in a dose dependent manner.After treatment with H2O2 of 0.5 mmol/L for 24 hours,the cell viability began to decrease in melanocytes with the expression level of DJ-1 being 2.23 times that in the untreated melanocytes(both P ＜ 0.05).Compared with the mock control group,the dendrites of melanocytes in DJ-1 group were obviously shortened with cytoplasm vacuolization.After 24-hour treatment with H2O2 of 0.5 mmol/L,the cell viability in the DJ-1 group dropped to 35％ of that in the mock control group(P ＜ 0.05),while the intracellular ROS fluorescence intensity( FI) and apoptosis rate were higher in the DJ-1 group than in the mock control group (902 ± 40 vs.529± 32,58q％ ± 6.1％ vs.30% ± 3.8％,both P ＜ 0.05).Conclusion DJ-1 can protect melanocytes against H2O2induced oxidative stress likely by decreasing intracellular ROS production and inhibiting ceU apoptosis.