Objective To explore the effects of octylphenol, an environmental contaminant with estrogenic activity, on the expression of apoptosis regulator gene bcl-2 mRNA in human breast cancer cell line MCF-7 and to compare its effects with 17?-estradiol (E 2). Methods MCF-7 cells were cultured in vitro and exposed to various concentrations of octylphenol and E 2 from 12 h to 120 h respectively. Total RNA was abstracted and the expression of bcl-2 mRNA was measured using reverse transcription polymerase chain reaction (RT-PCR) approach. Results The expression of bcl-2 mRNA in MCF-7 cells increased significantly after exposure to octylphenol and E 2 24 h and the effects continued to 120 h. The stimulation effect on the expression of bcl-2 mRNA was induced by octylphenol with a wide range of concentrations, and the strongest effect was found at the concentration of 10-6 mol/L of octylphenol. Conclusion The results suggested that octylphenol had similar estrogenic activities as E 2 on regulating the expression of bcl-2 mRNA in MCF-7 cells, but the effect was weaker than that of E 2.

Hyperplastic scar and contracture are two mainly respects that contribute to poor functional recovery of the patients after burn.The principles of prevention and management of the scar were reviewed in the article and the methods in facilitating functional recovery were also discussed as well.

Objective To investigate the effects of hepatitis B virus X (HBX) gene on cell morphology and transdifferentiation of human proximal tubular epithelial cells.Methods The eukaryotic vector pcDNA3.1-myc-HBX containing HBX gene was transiently transfected into HK-2 cells by lipofectamine mediation.The expression of HBX was confirmed by Q-PCR and Western blotting.Untransfected HK-2 cells and those transfected with empty vector were used as control.The morphology of HK-2 cells was observed by microscopy,the expressions of differentiation marker proteins α-SMA and E-cadherin were detected by Western blotting and Q-PCR,and the contents of IL-1,IL-6 and TNF-α in the supernatant were detected by ELISA assay.Results HBX was successfully expressed in HK-2 cells after transfection.After transfection of HBX gene,the shape of HK-2 cells became irregular,HK-2 cells significantly expressed E-cadherin and α-SMA,and had high levels of IL-1,IL-6 and TNF-α in the cell supernatant (P＜0.01).Conclusion Overexpression of HBX gene in renal tubular epithelial cells may damage cell morphology and promote the occurrence of epithelial-mesenchymal transdifferentiation,which may be related to the inflammatory microenvironment.

Objective To investigate the effect of hepatitis B virus X (HBX) gene on apoptosis and immune molecules of human proximal renal tubular epithelial cell line (HK-2).Methods The eukaryotic vector pcDNA3.1-myc-HBX containing HBX gene was transiently transfected into HK-2 cells by lipofectamine mediation.Untransfected HK-2 cells and those transfected with empty vector were used as controls.The TLR4 expression was detected by real-time PCR and Western blotting.The apoptosis of cells and expression of MHC-Ⅱ and CD40 were detected by flow cytometry,and the contents of IL-4 and IFN-γ in the supernatant were detected by EIISA.Results Compared with control groups,the number of apoptotic cells was significantly increased in the HBX transfection group (P ＜ 0.05),and the expressions of TLR4,MHC-Ⅱ and CD40 were also significantly increased in the HBX transfection group (all P＜0.05).IFN-γ level in the supernatant of HBX transfection group was higher (P ＜ 0.05),but IL-4 level was lower as compared to control groups (all P ＜ 0.05).Conclusions Over-expression of HBX gene may induce apoptosis of HK-2 cells and upregulate the expression of immune molecules of renal tubular epithelial cells leading to injury of cells and dysfunction of immunomicroenviroment.

The content of bicarbonate ( HCO-3 ) and carbonate ( CO2-3 ) ions in groundwater and lake water reflects a broad set of carbon cycling reactions associated with decomposition or synthesis of organic compounds with mineral dissolution or precipitation, which indicates the local geochemical environment. However, the content of HCO-3 and CO2-3 changes easily under the influence of pH, temperature, atmosphere pressure in the process of sampling, transportation and storage, so it has been a worldwide problem to determine the real content of HCO-3 and CO2-3 ions in groundwater and lake water. This article proposed a new way to solve the problem by fast field detection of HCO-3 and CO2-3 ions through the use of pH electrode combined with carbon dioxide electrode. Studies showed in the base solution of pH=4. 8 ± 0. 1, the detection range of HCO-3 ion was 0. 027-570 mg/L and that of CO2-3 was 1. 25 ×10-8-39. 7 mg/L. In the most case, the coexisting ions and weak acid ( K+, Na+, Mg2+, Cl-, SO2-4 <100 mg/L;HSO-3 , NO-2 , HOAc<50 mg/L) did not interfere with the analysis. The method was validated for real water samples and the recoveries were in the range of 95. 2%-99. 2% with the relative standard deviations (RSDs) of 2. 6%-3. 7%. Compared with the acid-base titration method, the accuracy of this method had proved to be good. However, the method could be affected by temperature, so the standard solution and samples should be measured at the same temperature. Above all, this method is suitable for fast field analysis for HCO-3 and CO2-3 ions in the nature water as it is sensitive, fast, economical, and the electrodes are easy to carry and operate. It has been successfully applied in the determination of HCO-3 and CO2-3 in groundwater and lake water in Qinghai Province. Experiment showed that the pH of the groundwater samples from Haidong district was 6 . 4-7 . 4 , with 234-4096 mg/L HCO-3 and 0. 16-1. 89 mg/L CO2-3 . The pH of the lake water samples was about 8. 7, with 1. 36-1. 86 g/L HCO-3 and 32. 3-43. 9 mg/L CO2-3 , which was consistent with the previous results.

Objective To explore the difficulty and strategy of percutaneous transheptic stenting for the junction of malignant hepatic duct obstruction. Method Twenty-three patients with highly malignant biliary obstruction received percutaneous transheptic biliary drainage (PTBD) by placement of stent and/or internal-external drainage tube. All the 23 patientws were followed up to determine the successful rate of surgery, the incidence of complications, decreased serum level of bilirubin, patency time of biliary stent and survival time. Results The initial operation was sucessful in 22 cases, accounting for 95.65%. Twenty-one patients had significant decline in total bilirubin (60%) and 2 had not (10%). The levels of alanine aminotransferase (GPT) and aspartate aminotransferase (GOT) decreased markedly and returned to nomral in an average of 18 d. After operation, 2 patients had fever,1 biliary tract bleeding, 6 pain in the right upper abodminal region, 1 localized peritonitis around the puncture point, 3 GPT increase and 1 drainage tube slip. There were no serious complications such as hemorrhea and biliary fistula etc. The symptoms of all these complications disappeared in 2 to 5 days through haemostasis, anti-inflammatory therapy and liver function protection. The median survival time was 8.5 months. Conclusion Although the merging of malignant hepatic duct obstruction stenting is difficult, try to pass through the obstruction by guide wire and master the right methods of operation can significantly improve the successful rate of surgery.

Adult ; China ; Female ; Humans ; Male ; Middle Aged ; Program Evaluation ; Smoking Cessation ; psychology ; statistics & numerical data ; Young Adult

Objective To investigate the expression of Notch 1 receptor in renal tissues of patients with hepatitis B virus associated-glomerulonephritis (HBV-GN) and its role in the pathogenesis of HBV-GN.Methods A total of 48 patients with HBV-GN confirmed by renal biopsy during 2008-2010 were enrolled in the study.Distribution of Notch1 receptor in renal tissue of HBV-GN was detected by immunohistochemistry and the association between the distribution of Notch1 receptor and HBsAg was examined by double-label immunofluorescence assays.Correlations of Notch1 receptor expression with renal pathology and clinical parameters of HBV-GN were analyzed.Results Notch1 receptor distributed mainly in renal tubular epithelial cells and interstitial area as brownish red granules,and a few expression in glomerulus was also found.The positive score of Notch1 receptor expression in HBV-GN patients was significantly higher as compared to primary glomerulonephritis patients with serum HBsAg positive or negative and normal renal tissue controls.Notch1 receptor expression was more obvious in membrano-proliferative glomerulonephritis (MPGN) and mesangial proliferative nephritis (MsPGN) patients,but there was no significant difference among the different pathology groups.Distribution of Notch1 receptor was consistent with the distribution of HBsAg and its intensity was positively correlated with renal interstitial fibrosis (r=0.473,P=0.001),tubular atrophy (r=0.690,P=0.000),inflammatory cell infiltration (r=0.616,P=0.000).Negative correlation was found between renal function and the intensity of Notch1 receptor (r=-0.393,P=0.006).Conclusions Notch1 receptor expression increases in the renal tissues of HBV-GN patients and distributes mainly in renal tubular epithelial cells and interstitium,which is consistent with the distribution of HBsAg.Its intensity is closely correlated with renal interstitial lesions and renal function.Abnormal expression of Notchl receptor in renal tissue of HBV-GN may be involved in the progress of HBV-GN.

ObjectiveTo explore the role of ERK1/2 in the expression of the type-1 plasminogen activator inhibitor(PAI-1) induced by parathonnone (PTH) in human renal tubular epithelial cell line HK-2 cells.MethodsVarious concentrentions of PTH and manifold durations were applied in the test.The expression of PAI-1 mRNA and protein in HK-2 cells was measured by RT-PCR and Western blotting,respectively.Besides,ERK1/2 protein was detected by Western blotting before the ERK1/2 inhibitor incubated with the HK-2 cells or after.Results The expression of PAI-1mRNA and protein was gradually up-regulatad along with the increasing concentrations of PTH(10-12-10-10 mol/L).The maximum level of PAI-1 mRNA and protein was detected in 10-10 mol/L PTH and was 4.01 and 3.81 times of control group.Otherwise,the decreased expression of PAI-1 was found while the concentrations of PTH were beyond 10-10 mol/L.The levels of PAI-1 mRNA and protein were increased in pace withtime from 12 to 72 hour,in time-dependent manner,which was 4.06 (12 h) and 4.03 (72 h) times of 0 hour group.The levels of ERK1/2 and PAI-1 were ascended after 10-10 mol/L PTH incubated with the HK-2 cells (all P＜0.01).Howerver,both of them decended after cells were pretreated by the ERK1/2 inhibitor (all P＜0.01),but were still higher than those of control group(all P＜0.05).ConclusionERK1/2 kinase system partly participates in the regulation of PAI-1 induced by PTH in HK-2 cells.

Objective To explore the expression and role of Toll receptor 4(TLR4)in human proximal tubular epithelial cell line HK-2,infected by HBV. Methods The serum of HBV DNA copies between 107-108/ml was collected. Before and after infected by HBV DNA positive serum. the HK-2 cells' morphology and the expression of α-smooth muscle actin(α-SMA)were observed by microscopy and immunofluorescence, and the effects of different concentrations of lipopolysaccharides(U)S.TLR4-stimulating factor)and CLI-095(TLR4 Inhibitor)on the proliferation rate of HK-2 cells were observed by MTT assays. After HBV serum and 10μg/ml LPS and 5μl/ml CLI-095 acted on HK-2 cells,TLR4 protein expression was measured by immunofluorescence and Western-blotting assay, and HBsAg and HBeAg in cell culture medium were detected by ELISA. and HBV DNA copies by fluorescence quantitative PCR. Results The longer HBV infected HK-2 cells, the more irregular of the cells' shape, the fewer number of the cells were left. But compared with HBV infected after 24 hours, α-SMA was more expressed after HBV infected 12 hours. After infected by HBV serum in 24 hours.HK-2 cells' proliferation rate was positively correlation in a dose range of LPS, but was negatively correlated with the CLI-095(P＜0.05＝.The levels of HBsAg and HBeAg in cell culture medium were largest when the LPS concentration was at 10μg/ml and CLI-095 at 5μg/ml.The expression of TLR4 significantly increased in HK-2 cells treated with LPS compared with those with CLI-095.but HBV DNA levels and HBsAg and HBeAg expression levels were lower. Conclusions HBV infection may promote cell transdifferentiation and cell injury. The stimulation of HK-2 infected with HBV by LPS may upregulate the expression of TLR4 and reduce the copies of HBV DNA.