Objective To investigate the effects of ghrelin on colonic motility in mice.Methods The eflfects of ghrelin on colonic propulsive movement were detected by charcoal suspension pushing test after injection of normal saline and different doses of ghrelin(20,50,100,200 ng/g).The effects of atropine,NG-nitro-L-arginine methylester hydrochloride(L-NAME)or D-Lys3-GHRP-6 on the changes of colonic propulsive movement caused by ghrelin(100 ng/g)were also investigated.In vitro,the effects of different doses of ghrelin(0.01,0.1,1,10μmol/L)on the spontaneous contraction amplitude of proximal colonic circular muscle strips were studied.Results Ghrelin significantly accelerated the colonic propulsive movement in dose-dependent manner,but the efiect was significantly inhibited in the presence of atropine,L-NAME or D-Lys3-GHRP-6(t=10.230,12.560,11.590,P＜0.05).Administration of ghrelin significantly increased the contraction amplitude of colonic circular muscle strips.but this effect was inhibited when the colonic circular muscle strips were pretreated by tetrodotoxin.ConclusionsGhrelin can accelerate colonic propulsive movement by activating growth hormone secretagogue receptor of cholinergic excitatory pathways and nitrergic nervous pathways in the enteric nervous system of colon.

Objective To detect the expression and function of ZNRF3 in different kinds of thyroid cancer tissues and cells.Methods Immunohistochemistry was used to detect the expressions of ZNRF3 protein in 35 cases of papillary thyroid carcinoma and 10 cases of poorly differentiated thyroid carcinoma.The expressions of ZNRF3 gene in TPC-1 and 8505C were detected by RT-PCR,and the cell lines were derived from papillary thyroid carcinoma and poorly differentiated thyroid carcinoma respectively.After silenced ZNRF3 gene expression with lentivirus,the proliferation ability of TPC-1 cells were detected with CCK-8,the invasion and metastasis ability of TPC-1 cells were detected with Transwell.Results According to results of immunohistochemistry and RT-PCR,the expressions of ZNRF3 in papillary thyroid carcinoma cells and tissues were higher than those in poorly differentiated thyroid carcinoma cells and tissues,the differences were statistically significant (4.83±0.44 vs.3.13 ±0.59,t =2.20,P ＜0.05;1.01±0.06 vs.0.21±0.04,t =11.80,P＜0.01).After ZNRF3 geng silencing,according to the results of CCK8,the proliferation ability of TPC-1 cells was significantly enhanced in 72 h,the difference was statistically significant (0.96 ± 0.10 vs.0.64 ± 0.05,t =3.19,P ＜ 0.05);and according to the results of Transwell,the TPC-1 cell's invasion (0.12 ± 0.01 vs.0.09 ±0.00,t =5.48,P＜0.01) and migration (0.22 ±-0.01 vs.0.17 ±0.01,t =4.58,P ＜0.05) also increased,the differences were statistically significant.Conclusion The expression of ZNRF3 in papillary thyroid carcinoma is higher than that in poorly differentiated thyroid cancer.ZNRF3 is tumor suppressor gene in the thyroid tumors.

Objective To investigate the correlation between the expression of STAT3 and MMP-2 in human pancreatic cancer,and to probe the mechanism by which STAT3 signal pathway regulates the expression of MMP-2 in pancreatic cancer cells.Methods Immunohistochemistry was used to detect the expression of STAT3,phosphorylated STAT3(p-STAT3)and MMP-2 in pancreatic cancer tissues of 34 cases and normal pancreatic tissues of 10 cases.Correlation between the expression of STAT3、p-STAT3 and MMP- 2 were statistically analyzed.Human pancreatic cancer cell lines SW1990 was cultured.AG490,an inhibitor of the upstream Janus kinase(JAK)of STAT3 was added into the culture medium.Electrophoretic mobility shift assay(EMSA)was used to detect STAT3 DNA-binding activity in SW1990 cells.Western blot was used to detect the expression of STAT3,p-STAT3 in SW1990 cells.In addition,the protein and mRNA expression of MMP-2 in SW1990 cells were determined by Western blot and RT-PCR,respectively. Results Immunohistochemistry revealed that the expression rate of STAT3,p-STAT3 were both higher in pancreatic cancer tissues than in normal pancreas tissues(P

Objective To observe the influence of inactive FRMD4A gene′s expression on the biological behavior of tongue cancer CAL-27cell. Methods FRMD4A-siRNA was transfered into CAL-27 cell by lipidosome, to the expression of FRMD4A-siRNA in CAL-27 cell after transfection was detect by qRT-PCR cell proliferation , was checked by CCK-8,the influence of inactive FRMD4A gene on cell cycle distribution of CAL-27 cell was assayed by flow cytometry. Results Compared with the control group, FRMD4A mRNA expression significantly reduced in FRMD4A-siRNA interfering group (94%) and the cell proliferation index decreased(P<0.05). The cell cycle arrested in G1 period (P<0.05). Conclusion FRMD4A-siRNA could effectively inhibit FRMD4A mRNA expression in tongue cancer CAL-27cell, impact the distribution of cell cycle, and reduce cell proliferation.

Objective To explore the prevalence of polycystic ovary syndrome (PCOS) in nurse of reproductive age and compare the characteristics of four phenotypic subgroups.Methods A cross-sectional survey was carried out in nurses aged 18-45 years in Renji Hospital in 2011.Questionnaire and anthropometric and biochemical assessments were made.Pelvic ultrasound evaluations were made and blood androgen levels were determined.Diagnosis of PCOS was based on Rotterdam 2003 criteria,consisting of anovulation/oligo-ovulation (ANOVU),clinical and/or biochemical hyperandrogenism (HA) and polycystic ovaries (PCO).Results There were 520 participants and finally 486 individuals finished questionnaire and androgen level determination.283 subjects were totally normal,48 suffered from PCO,129 HA,and 89 ANOVU.54 out of 486 women were diagnosed as PCOS,a prevalence of 11.1％.A significant difference exited only in age among four phenotypic subgroups (P＜0.05).There was no statistic difference in other parameters.Conclusion Establishing an explicit definition of each condition in PCOS criteria has important investigational implications and increase the comparability of published researches.Application of Rotterdam criteria is feasible for earlier diagnosis and timely intervention in order to prevent serious complications.

Objective To research the ophthalmic nosocomial infection ,and to explore ophthalmic nosocomial infection risk fac-tors .Methods Retrospective analysis medical record information of 122 ophthalmic patients ,who were residency and nosocomial in-fection ,from January 2003 to December 2012 in department of ophthalmology ,renhe hospital of three gorges university .multivariate logistic regression analysis was used to screen risk factors .Results ophthalmic nosocomial infection rate was 2 .47% (122/4 931) , the highest rate was of respiratory infections ,accounting for 56 .56% (69/122) .Multivariate Logistic regression analysis showed that ,ntact check ,invasive operation ,hospitalization days were the risk factors for nosocomial infections .Conclusion Ophthalmology inpatients often merge hospital infection ,hospital should strengthen the hospital infection management ,strict execution aseptic oper-ation ,reducing the occurrence of hospital infection .

OBJECTIVETo establish sleeve gastrectomy(SG) rats model of obese type 2 diabetes mellitus(T2DM) for the research of hypoglycemic mechanism.

METHODSNine male Sprague-Dawley (6-week-old) rats were fed with high-sucrose and high-fat diet for 4 weeks, developing diet-induced obesity (DIO) rats model. The rats were then randomly divided into two groups. Six rats of them underwent sleeve gastrectomy(SG) as the sleeve gastrectomy group[SGG, body weight (471.8±17.9) g] and the other three rats underwent a laparotomy and stomach manipulation as the sham operative group[SOG, body weight (467.0±8.4) g]. The body weight, caloric intake and peripheral blood concentration of total ghrelin of rats were recorded after operation.

RESULTSThe weight of all the rats declined progressively after operation. The weight of the rats in SOG began to rise on the 5th postoperative day(POD) and regain their preoperative levels on the average 22nd POD. However, the weight of the rats in SGG began to rise slowly from the 9th POD, but was still lower than that of SOG[(487.4±10.1) g] and preoperative levels[(471.8±17.9) g] on the 28th POD[(420.1±18.6) g](P=0.001). Average caloric intake of rats in SGG was significantly lower than that of SOG after operation, but there was no significant difference between the two groups(P=0.121). The ghrelin level of SGG showed a continuous decreasing trend after intervention, decreased by 17.4% compared with the preoperative level (1595.1±14.4 ng/L) on the 28th POD[(1316.8±14.8) ng/L]. The ghrelin level of SOG did not change obviously before and after operation and both groups differ statistically(P=0.000).

CONCLUSIONSA SG rat model is successfully established. This model can be used for the further study of mechanism analysis of T2DM resolution after surgery.

Animals ; Diabetes Mellitus, Type 2 ; pathology ; Disease Models, Animal ; Gastrectomy ; methods ; Ghrelin ; blood ; Male ; Obesity ; blood ; surgery ; Rats ; Rats, Sprague-Dawley

The epitope-G1 gene of Bovine ephemeral fever virus (BEFV) glycoprotein was synthesised by PCR and cloned into expression vector pPIC9K to construct recombinant plasmid pPIC9K-G1. Then the pPIC9K-G1 was linearized and transformed into Pichia pastoris GS 115. The recombinant P. pastoris strains were selected by a G418 transformation screen and confirmed by PCR. After being induced with methanol, an expressed protein with 26 kDa molecular weight was obtained, which was much bigger than the predicted size (15.54 kDa). Deglycosylation analysis indicated the recombinant G1 was glycosylated. Western blot and ELISA tests, as well as rabbit immunization and specificity experiments indicated that the target protein had both higher reaction activity and higher immunocompetence and specificity. The recombinant G1 protein could be used as a coating antigen to develop an ELISA kit for bovine ephemeral fever diagnosis.

A novel unsaturated polyphosphoester (UPPE) was devised in our previous research, which is a kind of promising scaffold for improving bone regeneration. However, the polymerization process of UPPE scaffolds was unfavorable, which may adversely affect the bioactivity of osteoinductive molecules added if necessary, such as recombinant human bone morphogenetic protein-2 (rhBMP2). The purpose of this study was to build a kind of optimal scaffold named UPPE-PLGA-rhBMP2 (UPB) and to investigate the bioactivity of rhBMP2 in this scaffold. Furthermore, the cytotoxicity and biocompatibility of UPB scaffold was assessed in vitro. A W1/O/W2 method was used to fabricate PLGA-rhBMP2 microspheres, and then the microspheres were added to UPPE for synthesizing UPB scaffold. The morphological characters of PLGA-rhBMP2 microspheres and UPB scaffolds were observed under the scanning electron microscopy and laser scanning confocal microscopy. The cumulative release of UPB scaffolds was detected by using ELISA. The cytotoxicity and biocompatibility of UPB scaffolds were evaluated through examining the adsorption and apoptosis of bone marrow stromal cells (bMSCs) seeded on the surface of UPB scaffolds. The bioactivity of rhBMP2 in UPB scaffolds was assessed through measuring the alkaline phosphates (ALP) activity in bMSCs seeded. The results showed that UPB scaffolds sequentially exhibited burst and sustained release of rhBMP2. The cytotoxicity was greatly reduced when the scaffolds were immersed in buffer solution for 2 h. bMSCs attached and grew on the surface of soaked UPB scaffolds, exerting well biocompatibility. The ALP activity of bMSCs seeded was significantly enhanced, indicating that the bioactivity of rhBMP2 remained and still took effect after the unfavorable polymerization process of scaffolds. It was concluded that UPB scaffolds have low cytotoxicity, good biocompatibility and preserve bioactivity of rhBMP2. UPB scaffolds are promising in improving bone regeneration.

To develop a parent-metabolite pharmacokinetic model for risperidone (RIP) and its major active metabolite (9-hydroxyrisperidone) and investigate their pharmacokinetics characteristics in healthy male volunteers, twenty-two healthy volunteers were orally given a single dose of 2 mg RIP. Plasma samples were collected in the period of 96 hours and concentrations of RIP and 9-hydroxyrisperidone were measured by a validated HPLC/MS method. CYP2D6 phenotypes were identified by the T1/2 of RIP and 9-hydroxyrisperidone according to the literature. Model structure identifiability analysis was performed by the similarity transformation approach to investigate whether the unknown parameters of the proposed model could be estimated from the designed experiment. Pharmacokinetics parameters were estimated using weighted least squares method, and the final pharmacokinetics model were tested and evaluated by Monte Carlo simulation. Eighteen volunteers were phenotyped as extensive metabolizers (EM) and four volunteers were identified as intermediate metabolizers (IM). The final model included central and peripheral compartment for both parent (RIP) and metabolite (9-hydroxyrisperidone) respectively. Model structure identifiability analysis indicated that the proposed model was local identifiable. However, if the ratio of RIP converted to 9-hydroxyrisperidone was assumed to be 32% in EM, and 22% in IM, the model could be globally identifiable. The predicted time-concentration curve and AUC(0-t), C(max), T(max) of RIP and 9-hydroxyrisperidone estimated by the established model were in agreement with the observations and noncompartment analysis. Rate constant of RIP conversion to 9-hydroxyrisperidone was (0.12 +/- 0.08) h(-1) and (0.014 +/- 0.007) h(-1) for EM and IM, respectively. Elimination rate constants of RIP were (0.25 +/- 0.18) and (0.05 +/- 0.23) h(-1) for EM and IM, respectively. Model validation result showed that all parameters derived from the concentration data fitted well with the theoretical value, with mean prediction error of most PK parameter within +/- 15%. The established model well defined the disposition of RIP and 9-hydroxyrisperidone simultaneously and showed large inter-individual pharmacokinetics variation in different CYP2D6 phenotype. The model also provide a useful approach to characterize pharmacokinetics of other parent-metabolite drugs.
Adult ; Cytochrome P-450 CYP2D6 ; physiology ; Humans ; Isoxazoles ; pharmacokinetics ; Male ; Models, Biological ; Monte Carlo Method ; Paliperidone Palmitate ; Pyrimidines ; pharmacokinetics ; Risperidone ; pharmacokinetics