Objective To investigate the effect of Ginkgolide B (BN52021) on lipopolysaccharide (LPS) induced pancreas microvascular endothelialv (MS1) cells, and to explore its molecular mechanism.Methods The optimal concentration and best time point of LPS inhibing MS1 cell survival and the optimal concentration of BN52021 increasing survival of LPS induced MS1 cells were determined by MTT.The mRNA and protein expression of adenylate cyclase (AC), phospholipase A2 (PLA2), phospholipase Cβ (PLCβ),protein tyrosine kinase (PTK) and G protein coupled receptor kinase (GRK) in platelet activating factor receptor(PAFR) signal pathway in MS1 cells were determined by real-time PCR and Western blot.Results It was showed that 10 μg/ml LPS for 24 h was the optimal concentration and best time point to induce the decrease of MS1 cells.50 mmol/L of BN52021 was the optimal concentration of increacing survival of LPS induced MS1 cells.After LPS induction, AC, GRK, PLA2, PLCβ, PTK mRNA expressions of MS1 cells were 4.02 ±0.14, 2.63 ± 0.03, 3.31 ± 0.12, 2.09 ± 0.08, 1.85 ± 0.07, which were significantly higher than those in control group (P ＜ 0.01).After BN52021 treatment, AC, GRK, PLA2, PLCβ mRNA expressions of LPS induced MS1 cells were 2.35 ±0.13, 1.17 ±0.14, 1.87 ±0.11, 1.65 ±0.10, which were significantly lower than those in LPS induction group (P ＜ 0.01).The expression of PTK mRNA was 1.83 ± 0.13, which was not significantly different from that in LPS induction group.Western blot showed that the levels of protein expression were consistent with those of mRNA expression.Conclusions BN52021 can down-regulate the up-regulated genes expression of AC, GRK, PLA2 and PLCβ in the PAFR signal pathway in LPS induced MS1 cells.

Objective To study the relationship of insulin-like growth factor-Ⅰ receptor(IGF-ⅠR)and carcinogenesis of gastric cancer.Methods The expression of IGF-ⅠR was detected in 40 cases of resected gastric cancer tissues and adjacent normal gastric mucosa by immunohistochemistry.The expression of IGF-ⅠR in gastric cancer cell lines(MGC803 and SGC7901)was detected by Western Blot.siRNAs targeted to IGF-ⅠR were designed,synthesized and transfected into MGC803 cells,the changes of IGF-IR protein level were detected by Western Blot at 48 h after transfection,the cell proliferation was examined by MTT,and then the growth curve was obtained.Results The positive rate of IGF-IR expression in gastric cancer tissues was 75.5%,significantly higher than that of adjacent normal tissues (25%,P＜0.01).The expression level of IGF-IR was related with TNM stage,lymphnode metastasis (P＜0.05),but not related with sex,age,histological differentiation,invasion depth of gastric cancer (P＞0.05).Intense expression of IGF-ⅠR was showed in gastric cancer cell lines.The inhibition ratio of IGF-ⅠR expression in sigNAl group were 89.80%±4.10%,the cell proliferation decreased to mininlunl level at the fifth day aftertransfection(by 29.0%±4.0%of mock-treated group),the cell number decreased by 21.15%±1.10%of mock treated group at the same time.Conclusions IGF-ⅠR is over-expressed in gastric cancer cells and can be effectively silenced by RNA interferes,therefore the growth of tmnor cell Was inhibited.Thus,it indicates that IGF-ⅠR may be a promising target for gene therapy of gastric cancer.

Objective To develop a method for simultaneous determination of 4 compounds in Zhizhu Pills by HPLC.Method Reversed-phase HPLC was used to determine the content of the compounds such as hesperidin,naringin,neohesperidin and atractylenolideⅠ.The column was symmetry C18(250 mm?4.6 mm,5 ?m) with temperature of 30 ℃.Methanol-0.095 %phosphoric acid solution served as the mobile phase by gradient elution.The wavelength of detection was set at 283 nm and 224 nm.The injection volume was 10 ?L and the flow rate was 1.0 mL/min.Results Hesperidin,naringin,neohesperidin and atractylenolideⅠin Zhizhu Pills can be isolated completely.Three batches of samples were determined and the recoveries of naringin,hesperidin,neohesperidin and atractylenolideⅠwere 98.81 %(RSD=1.11 %,n=6),100.63 %(RSD=1.90 %,n=6),99.32 %(RSD=1.44 %,n=6) and 99.65 %(RSD=1.53 %,n=6) respectively.Conclusion This method is accurate,reliable and with good reproducibility.It can be used for the quality control of Zhizhu Pills.

AIM: To explore the effect of receptor tyrosine kinase system mediated by phosphotyrosine phosphatase (PTP) on tau phosphorylation in rat hippocampus. METHODS: Pervanadate (PVN), inhibitor of PTP or inhibitor of glycogen synthase kinase-3 (GSK-3), LiCl were injected into rat hippocampus by stereotaxy technique. The level of tau phosphorylation was detected by Western blot and immunohistochemistry after 24 h of injection. RESULTS: PVN significantly inhibited tau phosphorylation at PHF-1 epitope and the inhibition of tau phosphorylation by PVN was stronger than that of LiCl (P

Objective To develop a new type of Raman-enhanced substrate for rapid detection of E.coli based on label-free surface-enhanced Raman scattering( SERS) technology.Methods Stober’ s improved method was used to prepare 360 nm silica ( SiO2 ) nanospheres.Prepared gold core-silver shell nanoparticles( Au@Ag) of different size were attached to 360 nm SiO2 to fabricate the nanocomposites ( SiO2-Au@Ag ) that were characterized by transmission electron microscopy (TEM) and UV-visucl light adsorption spectra (UV-Vis).PATP was detected to select SiO2-Au@Ag with optimal SERS effect.This optimal SiO2-Au@Ag was used to obtain the sensitivity of PATP and E.coli detection after a simple mixed culti-vation.Results TEM images showed that Au@Ag aggregated with the size of Au@Ag attached to 360 nm SiO2 .UV-Vis spectra indicated that the maximum absorption of Au@Ag and SiO2-Au@Ag had a red shift with the invrease of Au@Ag size.The experiment results suggested that detection sensitivity of PATP by SiO2-100 nm Au@Ag 10 -10 mol/L, while the lowest detectable E.coli concentration was 105 CFU/ml.Conclusion The 360 nm SiO2 binding with 100nm Au@Ag exhibits great potential for SERS applications.

Objective:To construct suitable vectors for the secretory expression of hirudin in Pichia pastoris.Methods:The α-facor-hirudin gene was amplified from pPIC9-hirudin by PCR and sub-cloned into PA0815.The multi-copy recombinant plasmid pA0815-(α-Hirudin)n was constructed.The recombinant was transformed into P.pastoris strain GS115 for induction expression and then the activity of secreted products was identified.Results:A new multi-copy vector pA0815-(αHirudin)n was successfully constructed and was capable of secreting recombinant hirudin efficiently,which was confirmed respectively by PCR and SDS-PAGE.The products possessed the activity of thrombin inhibitor.Conclusion:This result offers efficient P.pastoris stains for mass production of biological active hirudin.

Objective To compare the clinical effect of whole brain radiotherapy (WBRT) for brain metastases from lung adenocarcinoma between patients with and without epithelial growth factor receptor (EGFR) mutations.Methods A retrospective analysis was performed for 89 patients with brain metastases from lung adenocarcinoma who were treated in our hospital from August 2010 to May 2015.EGFR testing was performed in all patients.WBRT (6-MV external X-ray beam) was performed at 30 Gy in 10 fractions or 40 Gy in 20 fractions;for patients with ≤3 brain metastases, simultaneous integrated boost intensity-modulated radiotherapy was performed at 40-45 Gy in 10 fractions or 50-60 Gy in 20 fractions.The response rate, intracranial progression-free survival (IPFS), and overall survival (OS) were compared between patients with EGFR mutations and patients with wild-type EGFR.The Kaplan-Meier method was used to calculate IPFS and OS, the log-rank test was used for survival difference analysis and univariate prognostic analysis, and the Cox model was used for multivariate prognostic analysis.Results For these 89 patients, the overall response rate was 62%, the median IPFS was 7.0 months (95%CI:6.060-7.940), and the median OS was 12.0 months (95%CI:9.539-14.465).The univariate and multivariate analyses showed that the response rate was associated with Karnofsky Performance Scale (KPS) score and EGFR mutation status (P=0.009 and 0.035);KPS score and EGFR mutation status were significant prognostic factors for IPFS (P=0.048 and 0.000);KPS score and primary tumor control were significant prognostic factors for OS (P=0.000 and 0.031).Conclusions After WBRT for brain metastases from lung adenocarcinoma, the patients with EGFR mutations have a higher response rate and a longer IPFS compared with those with wild-type EGFR, but there is no significant in OS between the two groups of patients.

Objective To systematically evaluate the role of laparoscopic living donor hepatectomy in living donor liver transplantation (LDLT).Methods A systematic literature search was conducted on Medline-Pubmed,Embase,Cochrane Library to find studies on laparoscopic living donor hepatectomy for LDLT.All extracted data were analyzed using the RevMan 5 software.Results Ten studies with a total of 1 059 participants were included in this analysis.Laparoscopic donor hepatecomy (LDH) was associated with significantly less intraoperative blood loss [SMD =-0.39,95％ CI (-0.73,-0.05),P ＜ 0.05],lower peak level of postoperative total bilirubin [SMD =-0.24,95％ CI (-0.47,-0.01),P ＜ 0.05]and longer operative time [SMD =0.50,95％ CI (0.04,0.96),P ＜0.05] when compared with those operated with open surgery.On subgroup analyses,hospitalization stay decreased in patients who underwent LDLT with grafts obtained by complete living donor hepatectomy (LDH) and left lateral sectionectomy (both P ＜ 0.05).LDH was comparable to open surgery in donor complication rates and in-hospital cost (P ＞ 0.05).There were no differences on the harvested liver graft size,ischemic time,recipient postoperative liver function and complications between the two groups (P ＞ 0.05).Conclusions Laparoscopic hepatectomy in living donor is a safe procedure for graft-harvesting,which improved the clinical outcomes of the donor,liver graft and recipient in LDLT.It has also the advantages of reduced blood loss,low peak levels of postoperative total bilirubin and short hospitalization stay.

Objective:To construct pGEX-3X/hTSHa Escherichia coli(E.coli)expression system and prepare purified recombinant GST-recombinant human thyroid stimulating hormone(rhTSH)α protein.Methods:The complete coding sequence of hTSHα was obtained by RT-PCR with total RNA extracted from fresh chorial tissue as the template,and thereafter cloned into expression vector pGEX-3X by EcoRl and BamHI digestion.The recombinant plasmid was transformed into E.coli Mach1-T1 and then induced expression by WrG.The GST-rhTSHα fusion protein was identified by SDS-PAGE and its antigenicity was verified by a modified competitive ELISA.Results:A specific protein band of 36 ku,in accordance with predicted molecular weight,could be visualized in SDS-PAGE.As the result of ELISA,the recombinant GST-hTSHα protein can inhibit the intact TSH molecular binding with anti-TSHα antibody in a dose dependent manner.Conclusion:The cDNA of hTSHα was cloned and the recombinant expression vector pGEX-3X/hTSHα was constructed successfully.The recombinant GST-rhTSHα protein could be highly expressed in E.coli Machl-T1 and was approved of possessing antigenicity.

To evaluate the sealing potential of self-designed root canal filling material made of calcium phosphate cement (alpha-TCP/TTCP, CPC), the apices of root canals of six adult dogs were purposely perforated and enlarged up to the No 40 instrument. Then CPC was used to fill the root canal. Mean while either calcium hydroxide (Ca(OH)2) paste or hydroxyapatite (HA) paste was used as control. The animals were killed at 4, 12, 20 weeks postoperatively. The different materials about ways of apical closure, restoration periapocal tissues and adaptability to the dentinal surface were observed by histomorphology and scanning electron microscopic. This study revealed that CPC had excellent biocompatibility and adaptability to the dentinal wall. Its osteoconduction can promote the formation of calcific barriers and healing of periapical tissue. The apex can be closed completely. Compared with the control pastes it has advantages of ease of manipulation and better sealing capability. The results showed that CPC could be used as a root canal filling material for pupless teeth with open apex and destructive periapical tissue.
Animals ; Calcium Phosphates ; pharmacology ; Dogs ; Root Canal Filling Materials ; pharmacology ; Root Canal Therapy