Cholesterol was originally identified as an element of biomembrane with a key role in the regulation of the stability of membrane. Recent researches had revealed that mortality was inversely related to cholesterol levels, and hypocholesterolemia correlated with a high risk of complication. Evidence is now accumulating that low serum cholesterol levels appear to be an independent predictor of short-term mortality.

Objective To investigate the DNA methylation feature and DNA methylation regulation to its transcription and expression of O6-methylguanine-DNA methyltransferase gene (MGMT) in NaAsO2-treated HaCaT cells. Methods HaCaT cells were treated 72 hours at intervals and repeatedly by 3.13, 6.25,12.50, and 25.00 μmol/L NaAsO2, MGMT gene promoter region was amplified in the transcription initiation site - 329 - + 93 region by bisulfate-sequencing polymerase chain reaction (BSP), the mRNA transcription and the protein expression of MGMT was detected by real-time quantitative PCR and Western blotting. NaAsO2-untreated HaCaT cell was set as a blank control, and human epidermal squamous carcinoma cell strain A431 was set as a positive control. Results Among the groups of HaCaT cells treated with 3.13, 6.25, 12.50 and 25.00 μmol/L NaAsO2, the positive rates of the DNA methylation of promoter region in MGMT gene were 0.63%(l/160), 6.25% (10/160), 10.63%( 17/160) and 18.75% (30/160), respectively, and methylated CpG sites were mainly located in - 249--146 region relative to transcription start site. There was no DNA methylation in the blank control. There were significant differences between the blank control and the NaAsO2-treated cells (x2 = 76.687, P< 0.05). Average levels of MGMT mRNA were 1.518 31 ± 0.180 54, 1.425 22 ± 0.180 39, 1.014 54 ± 0.096 79 and 0.887 72 ± 0.020 00, respectively among the groups of HaCaT cells treated with 3.13, 6.25, 12.50 and 25.00 μmol/L NaAsO2, compared with the blank control cells(1.198 29 ± 0.159 97), there were significant differences(F = 37.359, P < 0.05). Average levels of MGMT protein were 1.174 47 ± 0.064 75, 0.848 83 ± 0.057 01, 0.471 63 ± 0.023 34 and 0.240 34 ± 0.014 43, respectively among the groups of HaCaT cells treated with 3.13, 6.25, 12.50 and 25.00 μmol/L NaAsO2, compared with the blank control cells (1.066 19 ± 0.061 24), there were significant differences(F = 20.687, P < 0.05). Conclusions Arsenic can cause CpC island hypermethylation in the promoter region of MGMT gene, which results in inhibited MGMT mRNA transcription and protein expression. It might be one of the important mechanisms of arsenic-induced skin lesion.

ObjectiveTo evaluate the antitumor effects of lymphocytes from hepatic porta lymph node co cultured with staphylococcal enterotoxin A(SEA)?MethodsLymphocytes isolated from lymph node of 5 HCC patients were co cultured with SEA. The proliferation and cytotoxicity of the lymphocytes were observed. Flow cytometer was used to detect the expression of CD3?CD4 and CD8 on the surface of the lymphocytes. TNF ? and IFN ? released into the culture medium were measured by the method of ELISA. The results were compared with that yielded from TILs of the individual patients. ResultsCultured in the presence of SEA, there was a sharp rise of T cells, especially CD8 + cells. Cytotoxic activity was increased. TNF ? and IFN ? were predominantly produced in the first week.ConclusionsLymphocytes from lymph node activated by SEA were armed with an increased antitumor efficacy.

Abstract? AIM: Through the establishment of penetrating keratoplasty model of rats, to detect the role and its mechanism of immature dendritic cells with IL-10 gene modified.? METHODS: Allogeneic penetrating corneal transplantation in rat model was performed. SD rats were randomly divided into positive control group, GFP-DC group, 8-DC and IL-10-GFP-DC group.At 3d before keratoplasty, the rats were given tail intravenous injection with the same amount of PBS, bone marrow 8-DC ( DC had cultured for 8d ) from donor Wistar rats, GFP-DC after 48h transfection and IL-10-GFP-DC.Rats were observed under slit-lamp for corneal graft cases every day, and recorded rejection index and corneal graft survival time.At 14d after keratoplasty, pathologic and immunohistochemical examinations were performed.?RESULTS:Compared with GFP-DC group and 8-DC group, corneal graft survival time of IL-10-GFP-DC group was significantly longer ( P<0.01 ); at 14d after keratoplasty, corneal opacity, edema, neovascularization and rejection index of IL-10 -GFP-DC group were significantly lower (P<0.01).Pathological examination showed that in the three experimental groups corneal inflammation was lighter than the positive control group without significant central graft neovascularization. Immunohistochemistry showed: compared to the positive control group, GFP-DC group and 8-DC group, CD4+, CD8+, CD25+, IL-2+, NK+and NF-κB+positive cells in IL-10-GFP-DC group were lower(P<0.01).? CONCLUSION: After donor -derived immature dendritic cells pretreated, corneal graft survival was significantly prolonged, successfully induced corneal transplantation tolerance. CD4+, CD8+, CD25+, IL-2+, NK+and NF-κB+positive cells are involved in corneal allograft rejection regulation, IL-10-GFP-DC may reduce CD4+, CD8+, CD25+, IL-2+, NK+and NF-κB+positive cell infiltration, inhibit corneal transplant rejection.

Objective The different distribution and clinical significance of mean platelet volume (MPV) in the healthy normoglycemic and impaired fasting glucose (IFG) individuals were discussed.Methods The 499 individuals including 184 male and 315 female,who had undergone health checks in Tianjin Huanhu Hospital during May and July 2012 were studied retrospectively.Average age is forty-five ( thirty-five to eighty).Subjects were categorized into four groups according to fasting plasma glucose ( FPG) levels:G1 (3.89 mmol/L≤FPG<5 mmol/L, n=125),G2(5 mmol/L≤FPG <5.5 mmol/L, n=121), G3(5.5 mmol/L≤FPG<6.1 mmol/L, n=142), and G4(6.1 mmol/L≤FPG<7 mmol/L, n=111).G1, G2, and G3 are defined as normal FPG groups and G 4 is defined as IFG group.Eighty-nine cases in the same age patients with type II diabetes mellitus group ( G5 ) were observed at the same time.Results The MPV increased with the increasing FPG levels in the following order:G1(8.62 ±0.77) fl, G2 (8.85 ±0.80) fl, G3(8.90 ±0.69) fl,G4(9.14 ±0.78) fl and G5(12.03 ±1.42) fl.MPV[(12.03 ±1.42) fl] of type Ⅱdiabetes mellitus group(G5) was higher than that in the IFG group (G4)[(9.14 ±0.78) fl] and normal FPG groups[G1(8.62 ±0.77) fl,G2(8.85 ±0.80) fl,G3(8.90 ±0.69) fl] (F=12.773,P<0.01);MPV of the IFG group [ ( 9.14 ±0.78 ) fl ] was significantly higher than that in normal FPG groups [ G1 (8.62 ±0.77) fl,G2(8.85 ±0.80) fl,G3(8.90 ±0.69) fl] (F=12.773,P<0.01 for G4 vs.G1 and G2, P<0.05 for G4 vs.G3) ;MPV in the high-normal glucose group (G3) [(8.90 ±0.69) fl] was obviously higher than that in the low-normal glucose group (G1) [(8.62 ±0.77) fl] (F=12.773,P<0.05);MPV was positively associated with FPG in normal FPG groups ,IFG group and type Ⅱ diabetes mellitus group (G1-3:r=0.22, P<0.05;G4:r=0.26, P<0.01;G5:r=0.29, P<0.01).Conclusions Significant difference of MPV was observed in population of different FPG levels.Especially, MPV in IFG group was evidently higher than that in normal FPG group and was positively associated with FPG levels.

0.05) except that IFN-? secretion of L-SEA group was lower than that of SEA group at 4th day (P

Objective　To study efficacy of 2,4-dibromo-5, 5-d imethylhydantoin in killing vegetative forms of bacteria and spores of B. subtil is var.niger, and efficacy of 2,4-dibromo-5,5-dimethylhydantoin in dest roying antigenicity of HBsAg. Methods　Neutralizer test and efficacy of so lution of 2,4-dibromo-5,5-dimethylhydantoin in killing vegetative forms of ba cteria and spores. Neutralizer test and efficacy of solution of 2,4-dibromo-5, 5-dimethylhydantoin in destroying HBsAg antigenicity in suspension. Resul ts　The killing rate of Staphylococcus aureus and E. coli. was 100 % a fter exposure to its solution containing 4 mg*L-1 and 2 mg*L-1 av ailable bromo after 10 min and 20 min. The killing rate of spores of Bacil lus subtilis var. Niger also was 100% after exposure to its solution co ntainin g 2 000 mg*L-1 available bromo after 30 min. Its solution containing 1 0 00 mg*L-1 available bromo with could destroy HBsAg in su spension for 5 min. Conclusions　2,4-dibromo-5,5-dimethylhydantoi n can effe ctively kill vegetative forms of bacteria and spores of B. subtilis var.ni ger, and can completely destroy the antigenicity of HBsAg in the water.

Objective To investigate the diagnostic value of thinprep cytologic test(TCT) and electronic colposcope(EC) examination in patients with subclinical human papillomavirus(HPV) infection(SPI) and cervical intraepithelial neoplasia(CIN),and explore its concordance with pathologic diagnosis. Methods A total of 1 125 females were examined with TCT,744 of whom diagnosed with abnormality by TCT or suspect of cervical lesions were examined with EC,and multiple punch biopsies were performed in 706 of those with abnomal EC images and suspected diseases. ResultsThe sensitivity of TCT,EC examination and the combined examination of TCT and EC for SPI and CIN were 88.74%,88.79% and 90.63%,respectively,the specificity 85.44%,80.38% and 96.41%,respectively,the positive predictive value 74.06%,68.90% and 91.10%,respectively,and the negative predictive value 94.19%,93.61% and 96.21%,respectively.By the combination of the two examinations,the specificity and positive predictive value were significantly increased(P

Objective To investigate the effect of NaAsO2 on the binding levels of methyl CpG binding protein 2(MeCP2),DNA methyltransferase 1 (DNMT1) and histone deacetylase 1(HDAC1) to the hypermethylation promoter region of MGMT gene in HaCaT cells,in order to provide a basis to deepen the interpretation of the role of arsenic poisoning mechanism.Methods HaCaT cells were treated repeatedly and interval with different concentrations of NaAsO2(3.13,6.25,12.50,25.00 μnol/L,respectively) for 72 h.Untreated HaCaT was used as blank control group and human epidermal squamous carcinoma cell line(A431 cells) as positive control group.The binding levels to the two transcription regulatory regions(ChIP1,ChIP2) and to the coding region(ChIP3) of MGMT 8ene were detected by chromatin immuno-precipitation combined with quantitative PCR.Results The differences of binding levels of MeCP2,DNMT1 and HDAC1 to ChIP1 and ChIP2 in each group were significant (F=7.387,84.634,78.442 and 19.263,69.649,26.546,all P ＜ 0.05).The binding levels of MeCP2,DNMT1 and HDAC1 to ChIP1 and ChIP2 in each NaAsO2 exposed group[3.13 μmol/L NaAsO2 exposed group:(136.00 ±16.97)％,(145.00 ± 2.83)％,(88.50 ± 19.09)％ and (106.50 ± 37.48)％,(112.34 ± 8.73)％,(59.71 ± 8.49)％;6.25 μmol/L NaAsO2 exposed group:(130.00 ± 42.43)％,(154.50 ± 4.95)％,(101.00 ± 1.27)％ and (88.50 ±3.54)％,(134.32 ± 2.82)％,(102.75 ± 19.91)％ ; 12.50 μmol/L NaAsO2 exposed group:(141.50 ± 23.33)％,(161.50 ± 7.78)％,(125.00 ± 11.31)％ and (119.50 ± 24.75)％,(171.59 ± 3.54)％,(167.61 ± 10.61)％; 25.00μmol/L NaAsO2 exposed group:(134.50 ± 43.13)％,(472.50+ 50.20)％,(383.50 ± 30.41)％ and (180.09 ±12.73)％,(348.50 ± 27.58)％,(158.45 ± 12.02)％] were higher than that in the blank control group[(51.50 ±9.19)％,(82.00 ± 12.73)％,(25.03 ± 2.91)％ and (37.02 ± 4.24)％,(91.56 ± 26.16)％,(19.09 ± 2.90)％,all P ＜ 0.05].The differences of binding levels of MeCP2 to ChIP3 in each group were not significant(F =1.670,P ＞0.05),but the differences of binding levels of DNMT1 and HDAC1 to ChIP3 were significant (F =4.404,9.863,all P ＜ 0.05),and only the binding levels in the 25.00 μmol/L NaAsO2 exposed group [(615.85 ± 29.63)％,(306.09 ± 59.40)％] were higher than that in the blank control group[(99.70 ± 12.02)％,(92.45 ± 48.79)％,all P ＜ 0.05].Conclusions MeCP2 can bind to the methylated MGMT gene transcriptional regulatory regions which are induced by arsenic and leads to histone deacetylation by the recruitment of DNMT1 and HDAC1 and,meanwhile,DNMT1 can bind to the coding region of MGMT gene to recruit HDAC1 in a methyl DNA binding protein(MBD) independence manner and media MGMT gene silencing through the chromatin remodeling way,which might be the early molecular events of arsenic poisoning.

Objective To investigate the transcription and expression of DNA methyltransferase 1 (DNMT1) mRNA in endemic arsenism patients by burning coal usage,to probe its effects on the development and carcinogenesis of arsenism. Methods In 2008,68 arsenism patients(including 24 mild cases,28 moderate cases and 16 severe cases) were selected in the areas with endemic arsenism according to Standarding of Diagnosis for Endemic Arsenism from Xingren county,Guizhou province. Among the subjects,40 cases were diagnosed by pathological methods,and they were divided into general pathological changes(20),precancerous(14) and cancerous group(6). Tweleve kilometer away from the endemic arsenism area,23 controls were selected in Daguoduo village (non-arsenism exposure). Under the principle of informed consent,blood samples were collected from individuals. The mRNA expression of DNMTI was detected by real-time quantitative reverse transcription polymerase chain reaction(FQ-PCR). At the same time,skin tissue samples were collected from the voluntary surgical treatment patients with endemic arsenism (total 61 cases,including 34 general pathological changes cases,21 precancerous cases and 6 cancerous cases) and from the control(15 cases). DNMT1 protein was detected by immunohistochemical method.Results Average level of DNMT1 mRNA were 0.221 83±0.595 09,0.246 11±0.509 79 and 0.389 27±0.411 33 respectively among mild,moderate and severe arsenism group. DNMT1 mRNA level of mild and moderate group were obviously lower than the control group(0.695 95±0.463 98,all P < 0.01). The mRNA average level of DNMT1 were 0.320 64±0.547 46,0.313 09±0.529 13 and 0.159 07±0.342 56 individually among general pathological changes,precancerous and cancerous group,which were obviously lower than the control group(0.695 95±0.463 98,all P < 0.05). The expression rates of DNMT1 protein in skin were 88.24%(30/34),100%(21/21) and 100% (6/6) among general pathological changes,precancerous and cancerous group were higher than the control group [0(0/15),all P < 0.01],and the extent of expression gradually increased with the aggravation of skin damage(r,= 0.740,P < 0.01). Conclusions DNMT1 participated in the development of the arsenism. High expression of its protein was an early event during the process of the arsenism. DNMT1 may be the new target markers for early diagnosis and treatment of arsenism.