AIM:To study the expressing variation of TNF-? and IFN-? mRNA in mouse splenocytes induced by H22 tumor cells derived heat shock protein gp96-peptide complexes in vitro,and to observe the morphologic change of H22 tumor cells which treated with the culture supernatant.METHODS:H22 tumor cells derived HSP gp96 was obtained by the techniques for protein extraction and purification and was identified by Western blotting method.The expression values of TNF-? and IFN-? mRNA in spleen lymphocytes were detected by semi-quantitative RT-PCR.The morphologic changes of H22 tumor cells induced by the culture supernatant were observed by laser scanning confocal microscopy(LSCM)and transmission electron microscope(TEM).RESULTS:Purified heat shock protein gp96 was identified by Western blotting.The expression value of TNF-? and IFN-? mRNA in activated spleen lymphocytes induced by gp96-peptide complexes was higher than that in control groups(P

Objective:To explore the availability for tumor-derived gp96 to induce specific CTL activity of splenocytes in vitro.Methods:gp96 purified by the techniques for protein extraction and identified by SDS-PAGE gel electrophoresis and Western blot method; CD8+T cell induced by gp96 and CTL activity detected by flow cytometry, immunofluorescence technic and CCK-8 assay.Results:gp96 was identified by SDS-PAGE and Western blot; Analysis with FCM showed that the number of CD8+T cells(nearly 70%) was obviously increased after pulsed with gp96-peptide complexes as compared with that of control groups(35%,26%, P

AIM: To promote the Clinical Data Inter-change Standards Consortium (CDISC) standard in clinical trials and promote the standardization of clinical trial data. METHODS: To combine the implementation guide of Analysis Data Model (ADaM) and common problems of actual data, and to introduce the application of analytical data model ADaM in the safety of bioequivalence trails of generic drugs. RESULTS: For different types of clinical trial data, according to various situations that may occur, a safety analysis data set that meets the standards was generated. CONCLUSION: Under the background of the continuous development of generic drugs in China and the low degree of standardization of clinical trial data, the use of CDISC standards in clinical research can promote the standardization of clinical trial data, and can also shorten the time of statistical analysis and accelerate the process of drug development.

Objective To explore the effect of cell apoptosis and oxidative stress on stressive liv-er injury after traumatic brain injury (TBI) in rats. Methods The model of TBI was duplicated by u-sing modified Allen's mehtods. Forty male Wistar rats were randomly divided into control group and groups at 6,12,24,48 hours after TBI. The serum levels of ALT and AST as well as the levels of superox-ide dismutase (SOD) and malandialdehyde in liver tissue were measured. The index of hepatocyte apopto-sis was detected through flow cytometer. Pathological changes of liver tissues were observed under light and electron microscopes. Results After TBI, the serum levels of ALT and AST were significantly in-creased, while malondialdehyde was increased and SOD decreased in liver tissues. The electron micro-scope showed that the index of hepatocyte apoptosis reached a peak at 6 hours after TBi. Aggressive inju-ries of the liver tissues were observed after TBI, showed by pathological observations. Conclusion Cell apoptosis and oxidative stress may be involved in the pathogenesis of stressive liver injury after TBI.

Objective: To evaluate the cytotoxic effect of PEM? induced by HSPgp96 on anti-tumor in vitro. Methods; PEM? separated from mice induced by thioglycolate were divided into three groups randomly: Culture medium in control; LPS-induced group; HSPgp96-induced group. The production of NO, the cytotoxic effect to H22 cells and the morphologic change of PEM? were investigated separately by enzyme method, MTT assay and scanning electron microscope. Results: In vitro, HSPgp96 can increased NO production from PEM? of mice and significantly enhance the cytotoxic effect of PEM? to H22 cells as well as LPS. Conclusion: HSPgp96 can effectively induce the cytotoxic effect of PEM? on anti-tumor in which NO is one of the capital effective molecules in vitro.

AIM:To investigate the immunomodulatory effect of astragalus saponin(AS)on macrophages and explore the mechanism of its immunomodulation.METHODS:By adding different concentrations of AS into cultured mouse peritoneal macrophages in vitro,the influence of AS on the synthesis of nitro oxide(NO)was detected by NO kit(enzymatic method).MTT assay was used to determine the cytotoxicity of macrophages induced by AS.The morphological changes of macrophages were observed under transmission electron microscope.LSCM and specificity fluorescent probe Fluo-3/AM were applied to observe the change of Ca2+ in macrophages induced by AS.RESULTS:AS significantly increased NO synthesis,enhanced the effects of mouse peritoneal macrophages on killing carcinoma cells.Cell surface projection exhibited multiplication,becoming thickening and growth longer via transmission electron microscope.Increase in intracellular Ca2+ in macrophages was also observed.CONCLUSION:AS enhances the immune function of macrophages by increasing NO synthesis and enhancing cytotoxicity.The increased intracellular Ca2+ may be the mechanism of immunomodulatory effect of AS.

Objective To prepare the chitosan-Mg membranes and to explore the biocompatibility of membranes with PC12 cells and its usage as biomaterials in tissue engineering.Methods The appearance of the chitosan-Mg membranes (CM) was observed under scanning electromicroscope (SEM) and the element of the membranes was analyzed by x-ray energy spectromete. PC12 cells were co-cultured with CM in vitro,the morphological changes of PC12 cells on membranes were observed under scanning electron microscope (SEM) and light microscope (LM),the cell vitality was detected by MTT assay. Results The surface of chitosan membranes (CS) was smoother than that of CM;but the CM groups were full of pore observed under SEM;The content of Mg element was related with the dosage of MgSO4 added into chitosan solution. Morphology observation showed that PC12 cells grew well on CM as compared with CS group. The cells were rich of microvilli and long processes on 7th day,and synapses-like structure was formed in many PC12 cells. In addition,the cell viability of experiment group was higher than that of control group(P

Objective To compare the serum estradiol level of gouty patients with healthy controls and to investigate whether estradiol upregulatesthe expression of the uric acid transporter ATP-binding cassette superfamily member 2 (ABCG2) of human renal tubular epithelial cells (HK-2)..Methods Serum of 16 male gout patients and 16 male healthy controls and their estradiol level were assessed with ELISA.The HK-2 cells wer cultured with different concentrations of estradiol for 24 hours or 48 hours.mRNA expression of ABCG2 was assessed by quantitative polymerase chain reaction (qPCR).HK-2 cells were cultured with estradiol or estradiol and inhibitor of PI3K/Akt pathway LY294002.mRNA expression of ABCG2 was assessed by qPCR,while the Akt,p-Ser473-Akt,p-Thr308-Akt and ABCG2 expression were investigated by Western blot.Data was analyzed using either the one-way analysis of variance or the t test.Results The level of serum uric acid in gout patients was [(547±18) μmol/L],which was significantly higher than that of healthy controls [(344±12) μmol/L),t=-5.395,P＜0.01].The level of estradiol in gout patients was (45±6) μmol/L,which was significantly lower than that of healthy controls [(100±8) μmol/L,t=9.375,P＜0.01].The mRNA expression of ABCG2 of 10-4 mol/L estradiol group was elevated after 24 hours or 48 hours (t=3.168,t=3.990;P＜0.01).In the group of co-stimulation withestradiol and LY294002,the ABCG2,p-Ser473-Akt and p-Thr308-Akt expression were down regulated compared to the estradiol group.Conclusion There is a significant correlation between serum estradiol and uric acid.Estradiol inducesthe expression of ABCG2 of HK-2 cells by activating PI3K/Akt pathway.

AIM: To investigate the CdTe quantum dots coated with MPA and explore its biocompatibility with living cells. METHODS: CdTe quantum dots coated with MPA were prepared in aqueous phase and MPA CdTe QDs were Characterized with TEM, fluorospectrophotometer and ultraviolet spectrophotometer. QDs were Modified with with avidin, purified and prepared as flurescent probe. LSCM was used to observe the expression of MHC Ⅱ antigen on PMφ cells, which was labeled by QDs. Cell culture and MTT assays were used to determine the biocompatibility of MPA coated CdTe quantum dots with the B-16 cells as target cells. RESULTS: The particle diameter of CdTe quantum dots prepared in aqueous phase was well distributed. They had good photological performance and greater stability after coated with MPA. MHC Ⅱ antigen on PMφ was labeled with the QDs-Avidin fluorescent probe showed great fluorescence intensity, which was easy to be detected by fluorescence microscope and LSCM. MPA CdTe QDs showed cytotoxicity when its density was very high, but they showed little cytotoxicity during the normal use of influence label density limit. CONCLUSION: MPA CdTe QDs can be used as new fluorescent lable as they are of even size, not easy to bleach or quench, have good photological performance and stability and good biocompatibility.

Objective To summarize the clinical diagnosis and treatment of nodular regenerative hyperplasia of the liver. Methods Retrospective analysis was made on the clinical manifestations,imagings, laboratory tests, diagnosis, treatment and prognosis of 18 consecutive cases finally established as NRH during the past 26 years. Results 15 of the 18 cases showed portal hypertension, 4 cases showed mono or multiple occupations of the liver, 8 cases suffered from concurrent autoimmune diseases, 3 cases were suspected of blood diseases. Preoperatively, 13 cases were diagnosed as cirrhosis, 2 cases were diagnosed as liver cancer or focal nodular hyperplasia ( FNH). All cases were diagnosed by operative wedging biopsy. 3 cases received splenectomy, 4 cases received disconnection /Phemister surgery, 3 cases received liver occupation/liver lobe resection, 1 case received partial small bowel resection, and 1 case received spleen artery restrictive surgery. Postoperatively, symptoms of portal hypertension relieved obviously. Follow-up study showed most of the patients were stable and prognosis of the NRH was good.Conclusions NRH may relate to the disturbance of liver blood supply, and most common clinical manifestation is portal hypertension, and can combine with immune diseases, hematopathy also can present single or multiple liver occupations. Differential diagnoses include liver cirrhosis, FNH, idiopathic portal hypertension. Diagnosis of NRH relies on liver wedging biopsy. Surgery can relive concurrent portal hypertension.