Epigenetics has become a hot topic in biomedical research in last a few years.Epigenetics refers to as epigenome,and it is interpreted as epigenetic changes (DNA methylation,histone modifications and non-coding RNAs such as microRNAs) in the regulation of genes expression on chromatin-based pathways rather than a single gene.Epigenetic factors such as DNA methylation,histone modification and miRNA serve as an interface for environmental stimuli and interacts each other to regulate gene expression and control phenotypes of cell.All of those epigenetic factors are essential in the keeping homeostasis of human body and contribute to many biologic conditions.In addition,the application of epigenetic drugs has been come into clinical trials especially in the treatment of cancers,therefore the understanding the mechanism of epigenetics in the development of human diseases and the therapeutic potential of epigenetic regulators in the treatment of human disease will bring us into the epigenetics era in biomedical research.

Objective:To explore the application value of MSCT in diagnosis of asymptomatic SPN through DR Chest Physical Examination.Methods: A retrospective analysis of 10 cases of complete image and pathological data of DR, CT, PET/CT confirmed by surgery in asymptomatic patients with SPN.Results: Solitary pulmouary nodules were diagnosed in the 10 patients with DR during chest examination. MSCT scanning indicated 8 cases of pulmonary cancers,one case of tuberculoma and one case of pulmonary inflammatory nodule. In 8 cases of pulmonary cancers, there found lobulation(7cases), spiculate(6cases), pleural invasion sign(5cases), vascular cluster sign(one case), and air-bronchogram(one case). PET/CT scanning testified 8 cases with above medium FDG uptake and 2 cases with mild FDG uptake. Surgical pathology showed 8 cases with lung adenocarcinoma,one case with tuberculoma and one case with granulomatous inflammation.The overall accurate diagnostic rate with MSCT was 100%. Tuberculosis and lung tumors in PET/CT examination showed above moderate FDG uptake, and one case of low FDG uptak was confirmed for pulmonary cancer by histopathology.Conclusion: MSCT is the most valuable imaging equipment in examination of asymptomatic SPN. Fine scanning with image post processing techniques is significant in the diagnosis of begnign and malignant asymptomatic SPN.

Objective To study the change of DNA polymerase beta and XRCC1's expression during malignant celI differentiation.Methods The Eca-109 cells were divided inm 2 groups:differentiation group which cultured with 8-Br-cAMP and control group.The 2 groups cells were cultured 48h simultaneously.The immunocytochemistry was performed to detect the expression of DNA polymerase beta and XRCC1.Results Compared with control group,the expression of DNA polymerase beta and XRCC1 was decreased simultaneously(P＜0.05).Conclusion The differentiation agent can down-regulate the expression of DNA polymerase beta and XRCC1,suggesting that overexpressed DNA polymerase beta and XRCC1 maybe result in mutator phenotype.

Aim To investigate the synergistic effects and possible molecular mechanism of hepatitis B virus (HBV) and alcohol on liver injury in HBV transgenic mice(HBV-Tg mice).Methods 20 HBV-Tg mice and 20 wild-type mice were randomly divided into 4 groups:alcohol-fed Tg mice and alcohol-fed Wt mice, and they were given intragastric administration with alcohol. Control Tg mice and control Wt mice received intragastric administration with saline.All groups were rasied for 10 weeks.The levels of ALT and AST in serum, the degree of inflammation, the degree of fibrosis, the mRNA expression of TGF-β_1, Smad3, Smad7, CTGF and the protein expression of TGF-β_1, CTGF, α-SMA in liver tissue were detected.Results The serumlevel of ALT and AST, the mRNA expression of TGF-β_1, Smad3, Smad7, CTGF and the protein expression of TGF-β_1, CTGF, α-SMA in liver all increased markedly in alcohol-fed Tg mice. Alcohol consumption induced hepatocyte steatosis and hepatic inflammation in alcohol-fed Tg mice, but the change of liver fibrosis was not remarkable.Conclusion HBV and alcohol have synergistic effects on early liver injury, possibly by enhancing the expression of TGF-β_1, Smad3, CTGF, α-SMA and inducing unbalanced expression of Smads.

Objective:To observe the role of lamividine and silymarin preventing and curing liver fibrosisrelevant factors induced by alcohol drinking in hepatitis B virus (HBV) transgenic mice (Tg mice).Methods:Forty HBV-Tg BALB/C mice with 1.3 copy were randomly divided into 4 groups:a control group,a model group,a lamivudine group and a silymarin group.Tg mice in control group were treated with normal saline via intragastric administration;Tg-mice in the model group were treated with 50％ alcohol (5 mL/kg) once a day via intragastric administration;while Tg-mice in lamivudine group and silymarin group were treated with alcohol (5 mL/kg) plus laminvudine (100 mg/kg) and silymarin (200 mg/kg) once a day via intragastric administration respectively.All groups were raised for 10 weeks.The levels of HBV-DNA copy number,ALT,AST in serum,the degree of inflammation,the degree of fibrosis,the mRNA expression levels of TGF-β 1,Smad3,Smad7 and connective tissue growth factor (CTGF),and the protein expression levels of TGF-β1,CTGF and α-SMA in liver tissue were detected.All the images were scanned with electronic computer and the data were analyzed with SPSS13.0 software.Results:Compared with the control group,liver injury were significantly aggravated,while HBVDNA copies,mRNA levels ofTGF-β1,Smad3,Smad7 and CTGF as well as the protein levels of TGF-β1,CTGF and α-SMA were significantly increased (P＜0.05).Compared with the model group,liver injury were significantly attenuated in silymarine group and lamivudine group,while mRNA levels of TGF-β 1,Smad3 and CTGF as well as the protein levels of TGF-β1,CTGF and α-SMA were significantly decreased;mRNA level of Smad7 was further increased (P＜0.05);the levels of ALT and AST in serum were decreased in the silymarine group (P＜0.05).Conclusion:Lamivudine and silymarin relieve the histological damage in the liver of alcohol-fed Tg mice.The mechanisms for the beneficial effects of lamivudine or silymarin might be related to inhibiting the expression of TGF-β 1,Smad3 and CTGF,modulating the expression of Smads and suppressing the activation of HSC.

OBJECTIVE: To establish a HPLC method for content determination of gatifloxacin in human plasma.METHODS: The Analytical column was C18,the mobile phase consisted of acetonitrile-30mmol/L ammonium acetate-triethylamine-orthophosphoric acid (20∶80∶1.0∶0.7) with a flow rate of 1.0ml/min,the detection was performed at UV 294nm.RESULTS: The calibration curve was linear in the concentrations ranging from 0.1 to10.0?g /ml (r=0.9 992).The detection limit was 0.05?g/ml.The intraday RSD was less than 8% and interday RSD was less than 10%.The average recovery was (101.67?3.06)%.CONCLUSION: The method is simple, sensitive, accurate and suitable for determination of gatifloxacin in human plasma and pharmacokinetic study.

Objective To reduce or prevent the incidence of bile duct injury in the laparoscopic cholecystectomy,and reduce the postoperative complications of Bile duct injury in the laparoscopic cholecystectomy by time and exact treatment during the surgical procedure.Methods We retrospectively analyzed the clinical data of 8700 cases of LC, including 12 cases of bile duct injury, from September 1991 to today. The corresponding treatment was used in LC to avoid bile duct injury, when bile duct injury occurred, the injury was recognized and immediately repaired by surgery. Result The mean follow-up period after surgery was 5-10 years,all 12 patients with bile duct injury were cured and discharged. Conclusions We should correctly handle the relationship of various duct and avoid burn or cut duct for reducing incidence of the extrahepatic bile duct injury. Correct treatment can reduce or avoid the incidence of postoperative complications.

Objective To establish a method of quantiying trace α-galactosidase from Bacteroides fragilis in enzymatic conversion of blood group B to O red blood cells ( B-ECO RBCs) .Methods BALB/c mice were immunized with purified recombinant B.fragilisα-galactosidase ( the purity>90%) to prepare monoclonal antibodies.The ascites were prepared using hybridoma cell lines stably secreting antibody and purified by HiTrap rProtein A column.The antibody titer and spe-cificity were detected by ELISA and Western blotting, respectively.Purified monoclonal antibody and rabbit polyclonal an-tibody were applied to detect residual enzyme in B-ECO RBCs and the washing solution was analyzed by indirect ELISA. Results A high titer and purity antibody was obtained.Western blotting showed that the antibody specifically reacted with B.fragilisα-galactosidase.Moreover, indirect ELISA was sensitive enough to detect the minimal amount of residualα-gal-actosidase at the concentration of 1 ng/ml.After four repeat washing cycles with 1∶4 ( v/v) phosphate-buffered saline, the amount of residual enzyme in B-ECO RBCs was less than 10 ng/ml.Conclusion An effective method of detecting the min-imal amount of residual α-galactosidase in blood conversion is established for safety evaluation of universal RBCs prepara-tion by enzymatic treatment.

Objective To investigate the role of microRNA-129-5p (miR-129-5p) in the regulation of epithelial-mesenchymal transition (EMT) of human peritoneal mesothelial cells (HPMCs) isolated from peritoneal dialysate effluents and TGF-β1 induced HPMCs line.Methods The isolated cells were cultured from peritoneal dialysate effluents overnight of 10 patients just started PD and 12 patients with PD over 6 months.Taqman PCR assay was used to determine the expression of miR-129-5p in the HPMCs.Moreover,the expression of miR-129-5p in HPMCs induced by 5 μg/L TGF-β1 for 0-72 h was also detected by Taqman PCR.HPMCs were pre-transfected with miR-129-5p precursor (pre-mir-129-5p) to overexpress miR-129-5p,then incubated with TGF-β1 for 48 h,and the expression of EMT associated gene and protein was detected by real-time PCR,Western blotting and immunofluorescence,respectively.Furthermore,the effect of TGF-β1 on the expression of Smad interacting protein-1 (SIP1) and the regulation of pre-miR-129-5p on the SIP1 expression also were investigated.Results MiR-129-5p expression significantly down-regulated in the HPMCs isolated from PD patients over 6 months than from PD start patients(P ＜ 0.01).Similarly,TGF-β1 remarkably decreased miR-129-5p in HPMCs lines on time-dependent manner (P ＜ 0.01).Pre-mir-129-5p dramatically restored the expression of epithelial marker E-cadherin,while inhibited the expression of Vimentin,a mesenchymal marker,in HPMCs induced by TGF-β1 (all P ＜ 0.01).In addition,TGF-β1 increased SIP1 expression in HPMCs time dependently,while the high level of SIP1 protein was obviously repressed after transfected of pre-miR-129-5p (P ＜ 0.01),but there was no obvious change of its mRNA expression.Conclusion MiR-129-5p modulates EMT formation of HPMCs in PD process,possibly by posttranscriptional inhibition of SIP1.Targeting miR-129-5p/SIP1 may provide a new approach for the prevention and treatment of peritoneal fibrosis during PD.

Objective:The study was attempted to improve the manipulation and eliminate patient's fear for gastroscopy combined with intravenous injection.Methods:A total of 1350 patients wasrandomized into test group (n=1100 given intraveous protofol and midazolam)and the control (n=250,without anesthetics).Patient's feeling,heart rate,blood oxygen saturation,blood pressure,operative duration and operator' satisfaction were recored and analysed.Results:Data in test group showed less complaints,easier manipulation,high satisfaction and no difference in operative duration and blood oxygen saturation as compared with the control.After intravenous administration of protofol and midazolam,patient's heart rate and blood pressure (systolic and diastolic pressure) significantly decreased.Conclusions:With intravenous use of protofol and midazolam,gastroscopy can be achieved effectively,painlessly and safely.