Aim To identify the GABA transporter subtypes and to find its new associated member at the blood brain barrier.Methods Labeled by in vitro infusion of magnetic beads through carotid arteries,the brain microvessels without intact neural cells were sorted in magnetic fields,and identified by RT PCR.The homologous primer of GAT superfamily and the tRNA from isolated brain microvessels were used in the RT PCR to amplify target DNA.The PCR products were isolated by polyacrylamide gel electrophoresis(PAGE) and cloned, sequenced rospectively.The sequences were screened in dbEST of Genbank by Blast.Results Seven DNA bands were isolated from RT PCR products of isolated brain microvessels by PAGE. B3,B5 complete sequences were highly homologous with rat partial GAT 2 and BGT 1 respectively,B7 complete sequence was highly homologous with rat partial TAUT. The other 4 EST of B1(Accession No:CF358965), B2(CD568346), B4(CF358966) and B6(CD568347) were submitted to dbEST,they were homologous with some sequences in Genbank,but were not homologous with GAT members.Conclusion GAT 2 and BGT 1 of GAT and TAUT were localized at the blood brain barrier which might be responsible for the GABA transport across the blood brain barrier.The genes and their functions of 4 EST associated with GAT need to be clarified.

Objective To evaluate the effect of early endoscopic treatment for patients with severe acute biliary pancreatitis.Methods Ninety patients with severe acute biliary pancreatitis were divided into three groups: Thirty patients underwent early endoscopic treatment(group A),30 patients underwent expectant treatment(group B) and 30 patients receive surgical treatment(group C),respectively.complications and safety were evaluated.Results The symptoms and signs disappeared in all 30 cases after early endoscopic treatment.All the 30 patients(100%) of endoscopic treatment(group A) were cured which significantly better than the other groups(group B 83.3% and group C 93.3%,respectively).Conclusions Early endoscopic treatment relieves the orifice obstruction of biliary and pancreatic ducts,decreases the pressure of biliary and pancreatic ducts,it is safe,mini-invasive and highly effective for the treatment of severe acute biliary pancreatitis.

Objective To observe the effects of oxaliplatin on proliferation in human hepatoma cell lines QGY in vitro and investigate the mechanism. To provide the theory foundation whether it can be used for the chemotherapy of hepatocellular carcinoma. Methods The inhibition of proliferation in QGY cell was estimated by MTT-test. Morphologic changes were observed under light microscope and electronic microscope. Distribution of cell cycle and apoptosis was analyzed using flow cytometry. The expression of cell cycle protein and apoptosis-associated gene protein was detected with immunohistochemical technique. Results Oxaliplatin could inhibit the proliferation of QGY cells and the inhibition depended on the exposure time and dose. The cells showed morphologic changes at the early stage of apoptosis under the light microscope: the shrunk and round cells, condensed cytoplasma and pycnosis nucleus. Apoptotic cells and apoptotic body could be found under the transmission electronic microscope. The analysis of cell cycle indicated that oxaliplatin blocked cells at S and G_2/M phases and the cells of G_0/G_1 phase reduced. When treated with oxaliplatin for 72 h, the expression of cyclin A and Bax were up-regulated, mutant type P53, Bcl-2 and Myc down-regulated, and Fas was not changed. Conclusion Oxaliplatin could inhibit proliferation of the hepatoma cell lines. Cell cycle blocked at S and G_2/M phase. The apoptosis were related to the up-regulation of Bax and down-regulation of mutant type P53, Bcl-2 and Myc. It could not induce apoptosis through the Fas approach.

Objective To evaluate the diagnostic value of CT in the spondylolysis of lumbar spine and improve the technique of CT scan.Methods The CT appearances of the spondylolysis of lumbar spine were analyzed in 20 cases.Results CT could demonstrate the spondylolysis and its abnormal features that led to compress nerve root.Conclusion CT scan plays an important role in the diagnosis of the spondylolysis of lumbar spine and in selecting treat methods.Technique of CT scan improved can depict the specific feature of spondylolysis truely.

Objective To investigate the effects and possible mechanisms of ursolic acid (UA) on inhibiting proliferation of human esophageal carcinoma cell Eca-109 and inducing its apoptosis. Methods Cell proliferation was determined by MTT assay. Electron microscope was used to observe the ultrastructural changes of Eca-l09 induced by UA. Cell cycle and apoptotic rate were analyzed by flow cytometry (FCM),and the expression of P27kip1,Bcl-2 and Bax were detected by Western blot method. Results UA could significantly inhibit the growth of Eca-109 cells(P

Objective To observe the effects of cytoplasmic transduction peptide (CTP)-HBcAg18-27-Tapasin induced murine bone marrow-derived dendritic cell (DC) maturation on T lymphocyte proliferation in vitro,Methods Bone marrow derived DC isolated from BALB/c mice were cultured with recombinant granulocyte-macrophage colony-stimulating factor and recombinant interleutin (IL)-4 for 5 days followed by lipopolysaccharide added to induce DC maturation.10 μg/L CTP-HBcAg18-27-Tapasin,50 μg/L CTP-HBcAg18-27-Tapasin,10 μg/L CTP-HBcAg18-27 or RPMI-1640 were added into culture medium to induce DC maturation.DC phenotypes were analyzed by flow cytometry.The level of IL-12p70 in the supernatant was detected by enzyme linked immunosorbent assay.The proliferation of.T lymphocytes was performed by using cell counting kit-8 and intracellular cytokine of proliferative T cells were analyzed by flow cytometry.The means among groups were compared using one-way ANOVA and those between two groups were compared by least significant difference test.Results DC were cultured and induced successfully.The molecules on DC surface,such as CD80,CD86 and major histocompatibility antigen-Ⅰ were upregulated by CTP-HBcAg18-27-Tapasin.IL-12p70 level induced by 50 μg/L CTP-HBcAg18-27-Tapasin was (61.12±10.25) pg/mL,which was higher than those induced by 10 μg/L CTP-HBcAg18-27-Tapasin (50.43±10.42) pg/mL,10μg/L CTP-HBcAg18-27 (40.17±8.54) pg/mL and medium control (30.51±8.03) pg/mL (F=15.85,P=0.030 and 0.037).The proliferation of T lymphocytes induced by CTP- HBcAg18-27 -Tapasin was higher than control groups.The amounts of cytotoxic T lymphocyte (CTL) induced by 50 μg/L CTP-HBcAg18-27-Tapasin [(2.05±0.41) ％] and 10 μg/L CTP-HBcAg18-27-Tapasin [(1.06 ±0.10 )％] were both significantly higher than the 10 μg/L CTP-HBcAg18-27 group [(0.45±0.11)％] and medium group [(0.09±0.02)％,F=60.22,P=0.003].Conclusions CTP HBcAg18- 27 Tapasin could promote the differentiation and maturation of DC,and enhance the ability of DC stimulating T lymphocytes proliferation and increase CTL expression effectively.

Objective To prepare Mi 2 antigen,detect anti Mi 2 antibody and investigate its significance in autoimmune connective tissue diseases (CTD).Methods Mi 2 antigen was prepared from rabbit thymus acetone powder and purified by DE52 chromatography.Anti Mi 2 antibodies were tested in 40 normal controls and 315 patients with CTD by double immunodiffusion.The distribution of anti Mi 2 antibodies in CTD was analysed.The clinical characteristics of dermatomyositis between anti Mi 2 positive group and negative group were compared.Results The precipitation line developed between the antigen and the standard anti Mi 2 serum.In DE52 chromatography,Mi 2 antigen was found in elute with 0 1 mmol/L and 0 2 mmol/L NaCl.The positive rate of anti Mi 2 antibodies was 26 1%(12/46) in dermatomyositis and 4 5%(2/44) in polymyositis respectively.There were no positive cases in 50 patients with rheumatoid arthritis,30 patients with primary Sjgren syndrome (SS),60 patients with systemic lupus erythematosus,50 patients with systemic sclerosis,35 patients with other CTD and 40 normal controls.The specificity of anti Mi 2 antibody in dermatomyositis was 99 4%.In dermatomyositis,the patients with positive anti Mi 2 antibody usually presented skin lesions at beginning,and most patients had V sign and/or shawl sign during the course of disease.On the other hand,the antibody negative ones manifested muscle involvements initially and easily got fever during the course of disease.There were statistic differences between the two groups in above features ( P

0bjective To observe the cell membrane penetration of protein transduction domain (PTD)-HBeAg fusion protein in vitro.Methods The sequence of trans-activator of transcription (Tat)-PTD was synthesized and the whole HBcAg gene was amplified by polymerase chain reaction (PCR).Overlap extension PCR was employed to fuse Tat-PTD and whole HBcAg gene.Then the fusion gene was cloned into prokaryotic expression vector pMAL-c2X.The correct vector was transformed into E.coli Rosetta-gamiTM 2(DE3),and the protein was induced by isopropyl β-D-1-thiogalactopyranoside(IPTG).Western blot was used tO identify the protein. Furthermore,the fusion protein PTD-HBcAg was purified by affinity chromatography.HBcAg protein expressed using the same methods was employed as eontr0l.The purified protein was added tO HuH-7 cell culture,then the transduction of PTD-HBcAg and HBcAg in cells were detected by indirect immunofluorescence assay (IFA).Results The fusion protein was effectively expressed in E. Coli and purified by affinity chromatography.Both purified PTD-HBcAg and HBcAg could be recognized by HBeAg monoclonal antibody in Western blot analysis.IFA visualization showed that PTD-HBeAg could be introduced into HUH-7 ceils while HBcAg only could not be detected in cells.Conclusions PTD-HBcAg fusion protein can be expressed effectively and purified in prokaryotic expression system.PTD could mediate HBcAg penetrating eell membrane into the cells.

Objective To determine the effect of different serum insulin-like growth factor-Ⅰ(IGF-1)levels on mouse cancer.Methods A total of 120 male mice at 6 weeks of age(60 control mice and 60 LID mice)were subcutaneously injected colon tumor CT26 cell line.Each group was random divided into two subgroups respectively,every 10 mice of one subgroup were injected subcutaneously with growth hormone(GH)(1ms/kg)daily from the 10th,14th and18th days respectively until the 22nd days,and the other subgroup received saline injection.Results All mice treated with GH have higher level of IGF-1,compared with those treated with saline.High level of IGF-1 promoted the development of cachexia in these mice treated with GH from the 10th days.However,the level of IGF-1 has negative correlation with the cancer cachexia state for mice treated with GH from the 14th days.Conclusion Circulating IGF-1 and GH play an important role in tumor growth 4nd cachexia development in the early stage of cancer and can ameliorate the state of cachexia in the advanced stage.

This article was to study the micro-leakage of 3 different root canal sealers (Endomethasone, AH-Plus and GuttaFlow) by fluid filtration test, and to observe the micro-structure between walls of root canal and the sealers by SEM. The results indicated that the micro-leakage of GuttaFlow was the least and Endomethasone was the most. Statistics difference were found between all the groups. Different root canal sealer got different micro-structure between walls of root canal and sealers. So we consider that the canal sealing ability of GuttaFlow is the best among these sealers.