Objective To study the absorption and transport mechanism of ursolic acid(UA) by using Caco-2 monolayer model.Methods Evaluating the transport characteristic through studying whether P-glycoprotein(P-gp) transporter inhibitor Verapamil(VER) exists or not by using Caco-2 cell monolayer as an intestinal epithelial cell model and investigating the effects of uptake time,drug concentration,system temperature,and pH value of culture media on the uptake of UA.The drug concentration was measured by HPLC coupled with UV detector.Transport parameters and apparent permeability coefficients (P_(app)) were then calculated.Results In the concentration range of 10—40?mol/L,the uptake of UA by Caco-2 cell all linearly increased.The P_(app) of UA transported on Caco-2 cell monolayer model significantly changed when the specificity inhibitory of P-gp was added to model and the apparent permeability ratio decreased from 3.445 to 1.386.Conclusion The intestinal absorption of UA is passive diffusion as the dominating process and active transportation mediated by P-gp transporter in Caco-2 cell monolayer model.

Objective To study and analyze the clinical efficacy of massage combined with Astragalus and eucommia Decoction in the treatment of periarthritis of shoulder.Methods100 patients with periarthritis of shoulder treated in our hospital from July 2015 to July 2016 were selected.They were randomly divided into the control group and the experimental group, with 50 patients in each group.Patients in the control group were treated with massage, and the experimental group was treated with massage combined with astragalus root and Eucommia ulmoides decoction.The 2 groups were treated with 2 or two courses, the 1 course of treatment for 15 days.The treatment effect and relative shoulder index score of the experimental group and the control group were compared and analyzed.ResultsAfter the corresponding treatment, the effective number of patients in the experimental group was 48 cases.The effective rate of treatment was 96%.The number of effective cases in the control group was 44, and the effective rate was 88%.The effective rate of the experimental group was significantly higher than that of the controlgroup, with statistical difference (P<0.05).After treatment, the shoulder function, activity and pain in the experimental group and the control group were significantly better than those before the treatment,The scores of shoulder functional performance index and shoulder pain index in the experimental group were significantly lower than those in the control group, with statistical difference (P<0.05).ConclusionMassage combined with Huangqi Eucommia Decoction in the treatment of periarthritis of shoulder can improve the therapeutic effect to a great extent, reduce the pain of patients with shoulder joint, to help with the recovery of shoulder joint function, with further clinical promotion and application significance.

To construct a cDNA library of gerbil′s cochlea, the mRNA was separated from the cochlea of gerbil and the first strand cDNA was synthesized through reverse transcription by a modified oligo(dt) primer. The double strand cDNA was amplified by Ld pcR. After cDNA size fractionation , the ds cDNA was ligated in the ? TripIEx2 vector . The cDNA library was identified with special primers of prestin genes of cochlea by PCR .The results showed that the titer of library was 1.8?10 6 pfu and the percentage of recombinant clones was 80% . Prestin gene was contained in the library , the size of the fragment was 863bp . The results suggest that the established cDNA library has a high titer, recombinant percentage and large insert fragments of genes . The study lays the foundation of molecular biological study of the cochlea.

Objective To investigate the optimal gestational weight gain (GWG) in twin pregnancies and to analyze the impact of GWG on pregnant outcomes.Methods A retrospective cohort study of twin pregnancies was conducted on women who gave birth in International Peace Maternity and Child Health Hospital,Shanghai Jiaotong University School of Medicine from January 1,2012 to the June 30,2015.An optimal range of GWG was calculated based on the amount of weight gain in 711 subjects who gave birth to normal twins and with uncomplicated pregnancy.Another 504 twin pregnant women without gestational complications were recruited for further analysis of the relationship between GWG and gestational outcomes.T-test,analysis of variance and Chi-square test were used for statistical analysis.Results (1) The optimal range of GWG for full-term twin-pregnancy was 15.3-21.4 kg.In those with low body mass index (BMI,＜18.5),normal BMI (≥ 18.5-＜25.0) and overweight/obesity (≥ 25.0),the GWG [M(P25-P75)] were 18.5 (15.8-22.2),18.3 (15.3-21.3) and 18.1 (14.9-21.5) kg,respectively.There was no significant difference in GWG among those groups (F=0.121,P=0.886).(2) According to the optimal GWG mentioned above,we divided the 504 cases into three groups,including lower GWG group (less than the optimal GWG,n=137),normal GWG group (in the GWG range,n=238) and higher GWG group (more than the optimal GWG,n=129).The neonatal birth weights in the three groups were (2 626.1 ±225.8),(2 680.14-237.9) and (2 751.9±257.1) g (F=9.189,P＜0.01),respectively,indicating that neonatal birth weight was increased by increasing GWG.The proportion of both twins with birth weights of more than 2 500 g was slightly increased,but there was no significant difference among the three groups [51.1 ％ (70/137),60.5％ (144/238) and 64.3％ (83/129),respectively,x2=5.279,P=0.071].The incidence of gestational diabetes mellitus (GDM) was reduced along with increased GWG [31.4％ (43/137),14.7％ (35/238) and 9.3％ (12/129),respectively,x2=25.144,P＜0.01],while the incidence of hypertensive disorders in pregnancy (HDP,including gestational hypertension and preeclampsia) in the three groups showed no significant difference.There were 90 cases (17.8％) of GDM in the 504 cases with a pre-pregnancy BMI of 22.5±2.8,which was higher than that of the non-GDM cases (21.21±2.9),(t=3.735,P＜0.01).Among the 504 cases,there were 67 cases (13.3％) of HDP (including gestational hypertension and preeclampsia) with a pre-pregnancy BMI of 22.4±2.8,which was higher than that of the non-HDP patients (21.3 ± 2.9,t=2.767,P=0.006).Conclusions The pre-pregnancy BMI has little influence on GWG in twin pregnancies.Increasing GWG to the recommended optimal range or above,and within a certain range,could promote an increase in neonatal birth weight without adding the risks of gestational complications,such as gestational diabetes mellitus,gestational hypertension and preeclampsia.

Objective To explore the absorption characteristics and mechanism of P-glycoprotein (P-gp) mediated transport of oleanolicacid (OA)across membrane in vitro.Methods The intake and transport of OA were evaluated by an HPLC/MS quantitative detection method.Safe concentration range of OA was determined by MTT.The effect of different drug concentration,incubation time,pH and temperature on the intake of OA by Caco-2 cells were investigated.The effect of P-gp inhibitor on the transmembrane transport of OA was investigated by the Caco-2 cell monolayer model,and the apparent permeability coefficient (Papp) was calculated.Results The uptake of O A was concentration-time dependent and correlated negatively to temperaturein Caco-2 cells within the safe concentration range,but was not significantly affected by pH value.Compared with controls,the Papp significantly changed when the P-gp inhibitor was added to the model(P ＜ 0.05).The apparent permeability ratio decreased from 2.90 to 0.95.Conclusion The absorption of OA was primarily by passive diffusion and might be mediated by the effiux by P-gp.

Objective To investigate the different expression pattern of Src-homology2 domain phosphatase (SHP)-1 and SHP-2 in human papiilomavirus (HPV)6/11 infected condyloma acuminatum (CA) and the significance of the difference. Methods HPV6/11 related CA cases were diagnosed by in situ hybridization. The expression and distribution of SHP-1 and SHP-2 were examined by SP immunohistochemistry technique in skin samples from 40 HPV 6/11 positive CA cases and 20 healthy control (foreskins). Results The positive rates of SHP-1 and SHP-2 were 80% and 85% respectively in CA, which were significantly higher than those in healthy control cases (only 35% and 30%, respectively, X2=11.87,P<0.01; X2 =18. 15,P<0. 01) . The SHP-1 and/or SHP-2 positive cells in CA skin lesions were mainly distributed in prickle layer, showing as brown yellow, with the positive staining located in cytoplasm. Contrastively, the SHP-1 and/or SHP-2 positive cells in healthy controls were rare and mainly distributed in basal layer, showing as pale yellow with the positive staining located in cytoplasm. There was no significant correlation between the expression of SHP-1 and SHP-2 in CA( rs = 1.0, P>0.05 ). Conclusion The expressions of SHP-1 and SHP-2 increase in HPV6/11 positive CA, which suggest that with the infection of HPV6/11, SHP-1 and SHP-2 may play a regulatory role in the proliferation of keratinocytes.

[Objective]This study was designed to explore the dynamic changes of cytokines in the local tissues of radiationinduced oral mucositis in a rat model.[methods]The rat oral mucosal tissues were obtained at 5 d,8 d,and 14 d after irradiation.which were received single X-ray irradiation of 30 Gy locally.RayBio~((R)) Rat Cytokine Array was applied to analyze the 19 cytokines.The mRNA levels of the 8 inflammatory-related cytokines were analyzed by real time-PCR.ELISA was employed to detect IL-1α,IFN-γ,and TIMP-1 protein levels.[Results]Cytokine array detection showed that cytokines such as Frac,IL-4,IL-6,IL-10,and TNF-α were down-regulated after irradiation.IL-1β,LIX,VEGF,and β-NGF were no obviously changed at the same time corse.The elevate range less than 2 fold of the CINC-2,GM-CGF,Leptin,MCP-1,and MIP-3α after irradiation were detected.The increasing range more than 2 fold of CINC-3,INF-γ,IL-1α,and TIMP-1 post irradiation were detected.The mRNA levels of anti-apoptosis,such as Bcl-2,and TNF-α,IL-6,VEGF,and IL-1β were low down,while the levels of promoting apoptosis cytokine such as Bax,and neutrophil recruitment associated cytokines,CINC,and TLR-9 were up-regulated after irradiation.The ELISA results of IL-1α,IFN-γ,and TIMP-1 were identical with the results of the two previom analyses.[Conclusion]The results of the three methods in the study should present evidences to prove each other.Some cytokines in the local lesion of oral mucositis during the initiate,development,and progression stages were blocked,while others were positively up-regulated to involving the control of the pathogensis of oral mucositis.

Objective To investigate the relationship between serum uric acid ( SUA) concentration and urinary albumin excretion rate ( UAER) and carotid intima-media thickness (CIMT) in patients with type 2 diabetes mellitua (T2DM). Methods The clinical data of 372 patients with T2DM. including 184 males and 188 females, were collected. The correlations between SUA and the other clinical indexes were analysed by Pearson method, and multiple regression analysis was employed to examine the effects of various factors on UAER and CIMT. Results SUA concentration was higher in males than in females with T2DM (P <0. 01). and was positively correlated with UAER both in males and females with T2DM, even after adjustment for the creatinine clearance (r = 0.24, P < 0.01 for males; r = 0. 29, P < 0.01 for females). Positive correlation was found between SUA concentration and CIMT in females (r =0. 29, P < 0. 01). Multiple regression analysis demonstrated that SUA concentration was an independent determinant of UAER for males as well as females (β=0.16, P<0.05 for males; β=0. 20, P < 0. 05 for females), and was also an independent determinant of CIMT for females (β =0.16, P <0.05). Conclusion SUA plays an important role in the progression of diabetic nephropathy and cardiovascular diseases in patients with T2DM. SUA control may provide a novel approach for the treatment of diabetic nephropathy and vascular complications.