Objective To explore the changes of hepatocyte proliferation in rats during the progression of nonalcoholic fatty liver disease (NAFLD). Methods Rat models of NAFLD were established by giving a fat-rich diet. The rats fed with normal diet were taken as normal control at the same stage during the study. These rats were sacrificed at week 4, 8, and 12 respectively and collected the liver tissues. The cell cycle and the changes of the S-phase cell fraction (SPF), proliferation index (PI) was measured by flow cytometry. The expression of proliferating cell nuclear antigen (PCNA) was detected by immunohistochemistry. Results There was no significant difference between NAFLD group and the control group at week 4, 8. But as compared with the control group, SPF and PI of 12-week group were significantly increased and PCNA labeling index were also higher. Conclusion Hepatic hyperplasia occurred after the nonalcoholic steatohepatitis was established in the rat models of NAFLD. Hepatic hyperplasia probably was the reaction of inflammation and necrosis, but whether it was the early incident of hepatocarcinoma should be paid attention.

Objective To investigate the curative effects of partial internal sphincterotomy in the posterior midline and lateral position in treatment of chronic anal fissure. Methods One hundred and fifty cases chronic anal fissure patients with stageⅡ or Ⅲ were randomly divided into observation group(80 cases)and control group(70 cases),who were hospitalized in the Shunyi District Hospital of Beijing from Dec. 2009 to Oct. 2012. The patients of two group were treatment by internal sphincterotomy either in the posterior midline or lateral position respectively. The efficacy,healing time,complications and recurrence of the two groups were recorded and compared. Results There were 76 cases with cure,6 cases with improvement and 2 with ineffective effect in the observer group. There were 49 cases with cure,12 cased with improvement and 9 cases with ineffective effect in control group. The difference of the efficacy was statistically significant( P =0. 025). Wound healing time of observation group and control group were(9. 9 ± 0. 8)d and(17. 1 ± 0. 9)d,and the difference was significant( P =0. 048 ). The incidence of anal exudates in observer group was lower than the control group(2. 5%(2/80)vs. 24. 3%(17/70);P=0. 032). The incidence of recurrence in observer group was 5. 0%(4/80),lower than the control group(24. 3%(17/70),P=0. 046). The degree of the postoperative pain,voiding dysfunction and wound edema in observer group were lower than the other group( P =0. 025, 0. 025,0. 049). Conclusion The effect of internal sphincterotomy in the lateral position in treatment of chronic anal fissure is better with quickly repair,fewer complications.

OBJECTIVE：To detetmine oinultaneous ly ephedtine hydrochlotide and diphenhydtamine hydtochlotiae without any sepatat ion in Antiasthmatic Mixtute.METHOD:Double-Wavelelength secondaty d erivative spectrophotometry and secondary derivative spectrophotometry were used .RESULTS:The content of ephedrine hydrochloride and diphenhydramine hydrochloride could be obtained respectively Linear ranges were both 5.0～25μg/ ml,the correlation coefficients were 0.9994 and 0.996,the average recoveries wer e 99.7% and 96.6%,RSD were 0.6% and 0.4%,respectively.CONCLUSION:The method was simple，rapid and accurate for routine analysis of hospital preparat in.

ObjectiveTo investigate the in vitro effect of ghrelin on membrane membrane potentail of GH3 cells.MethodsNystatin-perforated whole-cell recording configuration was used.ResultsThe membrane potentail of GH3 was decreased from-(72±4.78) mV to(54.8±4.58) mV(P<0.01) after application of ghrelin(10-7～10-8 M)ConclusionGhrelin can transiently and significantly decreased the amplitude of membrane potentail,which may contribute to the ghrelin-stimulated GH secretion from GH3 cells.

TLR4 mediates I/R injury involving endogenous ligands.Interaction of TLR4 with endogenous ligands provides a critical link between tissue damage and activation of the innate immune response.In the early phase of liver,kidney,heart,or lung I/R injury,endogenous ligands are secreted from several kinds of cells,they are recognized by TLR4.Interaction of TLR4 with endogenous ligands,such as HMGB1,seems to be the most important trigger of inflammation and initiates signaling cascades leading to inflammatory and immune responses.Blocking the interaction of TLR4 with endogenous ligands may be useful in clinical management of inflammation and cellular necrosis caused by ischemic insults.

Objective To prepare mouse polyclonal antibody against human Nanog by genetic immunization and to identify this antibody by Western blot and immunofluorescence. Method The antigenicity fragment (A16-V101) of human Nanog (hNanog) was chosen by analysis of Accelrys software, and its cDNA (258bp) was amplified from plasmid containing full-length cDNA of hNanog, then it was cloned into pBQAP-TT to construct recombinant plasmid pBQAP-TT-hNanog for genetic immunization. Mice were immunized with this recombinant plasmid and two other adjuvant plasmids-pCMVi-GMCSF and pCMVi-FIT3L, which help to enhance the antibody's generation. After 12 weeks, we obtained mouse anti-hNanog antibody from mice blood serum. The antibody titer was determined by enzyme-linked immunoadsordent assay (ELISA), and its specificity was identified by Western blot in human renal protein. Using this antibody, we detected hNanog expression in HKC cells of hNanog-AAV2 transfection. Results Recombinant plasmid pBQAP-TT-hNanog for genetic immunization was confirmed to be correct by restriction digestion and sequencing. The result of ELISA showed that the antibody titer was 1∶3 200. This antibody recognized a band of 34kD hNanog protein in human renal protein by Western blot. Immunofluorescence showed that Nanog protein was mainly located in the nuclei in hNanog transgene HKC cells. Conclusion Genetic immunization can offer mouse anti-hNanog polyclonal antibody of high titer and high specificity.

Objective To explore the changes of E-cadherin expression in renal ischemia-reperfusion injury.Methods For the in vitro analysis of epithelial ischemia,confluent monolayers of MDCK cells growing in DMEM were depleted of ATP for 4 h by incubation in PBS (supplemented with 1.5 mM CaCl2 and 2 mM MgCl2) containing 10 ?M antimycinA.For the in vivo studies of epithelial ischemic injury,adult Sprague-Dawley rats were subjected to bilateral renal artery ligation.Renal pathological changes were measured by PAS stain.Location and expression of E-cadherin were detected by immunohistochemistry and western blot respectively.Results E-cadherin were primarily found in a linear pattern at the lateral portions of the plasma membrane in normal MDCK.After ATP depletion for 4 hours,the linear pattern altered and manifested by the appearance of intracellular staining.In invivo ischemia-reperfusion model rats,E-cadherin expression was changed from normal tubular epithelial cell basal membrane to cytoplasma.Western blot suggested that in sham-operated rats,E-cadherin was 120 ku lane vs 80 ku lane in ischemia for 60 min rats,while in ischemia for 45 min rats,both the 120 ku and 80 ku lanes were detected.Conclusion In renal ischemia-reperfusion,the location and expression of E-cadherin are obviously altered in vivo and in vitro study and E-cadherin are degradated as ischemia time prolongs.These changes may be the reason why tubular epithelial cell exfoliated from TBM in ischemia-reperfusion injury.

Objective To compare the diagnostic value of magnetic resonance cholangio pancreatography (MRCP), endoscopic ultrasonography (EUS) and endoscopic retrograde cholangio pancreatography (ERCP) in obstructive jaundice. Methods MRCP, EUS and ERCP were performed in 39 patients with obstructive jaundice. MRCP was performed by the technique of T2 weighted fast spin echo fluid imaging. EUS and ERCP were performed with conventional method. Results The over all concordance rates of MRCP, EUS and ERCP were 87.2%(34/19), 94.9%(37/39) and 97.4(38/39), respectively. In malignant common duct stricture the concordance rates of MRCP, EUS and ERCP were 61.5%(8/13), 84.6%(11/13) and 92.3%(12/13), respectively. In choledocholithiasis the concordance rates of MRCP, EUS and ERCP were 100.0%(21/21). Conclusion The non invasive examination, MRCP can be the first choice as a diagnostic method for obstructive jaundice, but it can not replace ERCP currently. As an important approach to diagnose the cholangiopancreatic disease, EUS combine with MRCP and ERCP, can improve the diagnostic accuracy in obstructive jaundice.

Objective To understand the change of leptin level in human in acute hypoxia state. Methods 34 male mountaineers were examined, 7 at the altitude of 2 800 m, 27 at 5 050 m. The serum leptin level and anoxic degree were determined by radioimmunoassay and arterial oxygen saturation were determined by Tuffsat arterial oxygen saturation monitor. Results The leptin level at 5 050 m (9.08?7.77) ng/ml was significantly higher than that at 2 800 m(2.02?1.09) ng/ml, P

Objective To discuss the influence of drug compatibility on the components of the volatile oil in Flos Lonicerae and Fructus Forsythiae. Methods GC-MS system was used to analyze the volatile oil in Flos Lonicerae, Fructas Forsythiae and their combination. The effect of compatibility on the components and content of volatile oil was evaluated. Results The results of GC-MS showed that gerani]o, tetradecanoic acid methyl ester, hexadecanoic acid ethyl ester, fluoranthene, 9, 12, 15-Octadeeatrienoic acid ethyl ester and linoleic acid was detectable in Flos Lonicerae but was undetectable in the drug pair; camphene, alpha-terpinene, perillaalcohol, 2-beta-pinene, beta-hexadecanal, phellandrene was de-tectable in Fructus Forsythiae but was undetectable in the drug pair; limonene, isolongifolene, nonadecane was detectable in the drug pair but was undetectable in the two single drugs. Conclusion The compatibility of the drug pair has an effect on components and contents in the volatile oil of Flos Lonicerae and Fructus Forsythiae.