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 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.

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 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 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 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.

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 explore the effects of NF-E2-related factor 2 (Nrf2) overexpression on mitochondrial biosynthesis and function in melanocytes.Methods An immortalized human vitiligo melanocyte cell line PIG3V was used in this study.An overexpression plasmid Nrf2-pEX-1 containing the full-length Nrf2 gene was constructed.PIG3V cells were divided into 3 groups:blank group receiving no treatment,control group transfected with the pEX-1 plasmid,overexpression group transfected with the Nrf2-pEX-1 plasmid.After transfection,real-time quantitative reverse transcription-PCR (RT-PCR) and Western blot were performed to determine the mRNA and protein levels of mitochondrial biosynthesis-related factors (including Nrf2,nuclear respiratory factor 1 (NRF1) and mitochondrial transcription factor A (TFAM)) respectively;RT-PCR was also conducted to measure the copy number of mitochondrial DNA (mtDNA),and flow cytometry to estimate mitochondial membrane potential (MMP);luciferase reporter system was used to estimate the intracellular adenosine triphosphate (ATP) level.Statistical analysis was carried out by using a two-sample t-test.Results After transfection,a significant increase was observed in the mRNA expression levels of Nrf2 and NRF1 at 24 hours (both P ＜ 0.001) and in those of Nrf2 and TFAM at 48 hours (both P ＜ 0.05),but no significant change was noted in the mRNA expression level of TFAM at 24 hours (P ＞ 0.05) or in that of NRF1 at 48 hours (P ＞0.05) in the overexpression group compared with the control group.In the case of Nrf2,NRF1 and TFAM protein levels,the overexpression group showed significant increases compared with the control group at 48 hours after transfection (all P ＜ 0.05),while no significant difference was noted between the two groups at 24 hours.Compared with the control group,MMP in the overexpression group increased by 2.313％ at 24 hours (t =5.546,P =0.005) and by 14.872％ at 48 hours (t =8.537,P =0.001) after transfection.Both the relative copy number of mtDNA and ATP level were similar between the overexpression group and control group at 24 hours after transfection (both P ＞ 0.05),but significantly higher in the overexpression group than in the control group at 48 hours (t =5.760,P =0.005;t =22.040,P =0.008).Conclusion Up-regulation of Nrf2 pathway can improve mitochondrial function and biosynthesis in PIG3V cells likely by promoting the expressions of mitochondrial biosynthesis-related genes and proteins.