Objective To clone and express the gene encoding the binding domain of human soluble complement receptor type 1 (sCR1). Methods sCR1-SCR15-18 cDNA was amplified using RT-PCR from human monocytes of peripheral blood and sequenced using vector pMD-18T. Recombinant pET32-sCR1-SCR15-18 was constructed using prokaryotic expression vector pET32 and transformed into bacterium BL21. IPTG was used to induce gene expression and the obtained expression product was identified by immunoblotting. Results The gene segment that specifically encodes sCR1 was synthesized, the sequence of which was consistent with that of sCR1-SCR15-18 cDNA as registered at GenBank. A prokaryotic expression recombinant pET32-sCR1-SCR15-18 was constructed. The amount of target protein accounted for 40% of the total bacterial proteins and inclusion bodies were present in the bacteria. Immunoblotting showed a single positive band at the site of 43?10~(3). Conclusion The gene encoding sCR1-SCR15-18 was cloned from human monocytes and efficiently expressed in E.coli.

Objective: To observe the influence of spent culture supernatant (SCS) of Lactobacillus acidophilus strain LA14 on the contraction of isolated intestinal smooth muscle and to discuss the related mechanism. Methods: The ileum samples of rabbits were prepared and the contraction frequency and amplitude of intestinal smooth muscle were observed as the normal control. Then the SCS, bacterium suspension, and SCS with bacterium suspension were added by an accumulative dose to the culture media (0.3 ml per times, at an interval of 6 min), respectively. Four minutes after each administration, the contractive curves were recorded for 2 min. The influences of various groups of Lactobacillus acidophilus on the contraction of isolated intestinal smooth muscle were observed. The effect of SCS on M cholinoceptor was observed by adding in order pilocarpine, atropine or SCS, and pilocarpine. Results: After continuous administration of SCS or SCS with bacterium suspension (0.6-1.5 ml), the contraction frequency of the intestinal smooth muscle was significantly lowered compared with before administration (P<0.05 or P<0.01), and there were no significant differences between before administration and other volume groups (P>0.05). Within the range of 0.3-1.5 ml, the SCS, bacterium suspension, and SCS with bacterium suspension resulted in no significant difference in reducing the contraction amplitude, except for SCS with bacterium suspension at 1.5 ml(P<0.05). SCS or atropine significantly inhibited pilocarpine-induced increase of contraction amplitude(P<0.05 or P<0.01). SCS also reduced the contraction frequency of the intestinal smooth muscle(P<0.01). Conclusion: SCS of Lactobacillus acidophilus may inhibit the peristalsis of the intestinal smooth muscle of rabbits by blocking M cholinoceptor.

Kaposiform hemangioendothelioma ( KHE ) is a vascular tumor characterized by intermediate malignancy.Retrospective analysis and literature review on the clinical pathologic,immunohistochemical and clini-cal data of a patient with mesentery KHE.The cardinal symptom of this patient is alimentary track hemorrhage, imageological diagnosed as a huge soft tissue mass of mesentery and invaded ileum.Clinical feature is without Ka-sabach-Merritt phenomenon,observing under the light microscope.The tumor is with cavernous vascular structure, and lymphocyte infiltration inside of stroma.The nodules are composed of disposed and lots of short spindle cell tumor;the tumor cells inside of the nodules are vertical and horizontal staggered and form tendon like or fissuring vessels.Immunohistochemistry:CD31(+),CD34(+),D2-40(+),C-Kit(-),SMA(-),Ki-67(1%).

Objective To obtain the prevalence data on human T Lymphotropic virus (HTLV) infection in 2 339 blood donors of an endemic coastal region of Fujian, China. Methods Serum antibodies to HTLV in the donors were detected by a home made double antigen sandwitch ELISA kit. All the ELISA positive samples were further confirmed by Western blot (WB) and/or polymerase chain reaction (PCR) combined with sequencing. Results Nine samples were confirmed as HTLV 1 infection among the 2 339 donors. A man whose wife is one of the nine was also detected to infect by WB and PCR. Conclusion The prevalence of HTLV infection was 0.38% in the endemic coastal region of Fujian, China.

Objective To investigate the expression of TMS1/ASC gene induced by gemcitabine(GEM) in pancreatic carcinoma cell line PANC-1.To study the relationship between cysteine aspartase(caspase-1),nuclear factor-κB(NF-κB) and the expression of TMS1/ASC.Methods The pancreatic carcinoma cell line PANC-1 was cultured in Dnlbecco's modification of Eagle's medium(DMEM).Methyl thiazolyl tetrazolium (MTT)method was used to measure the effect of GEM at different time points(24,48 h)at different concentrations(1,2,4,8,16 μg/ml) on growth of PANC-l.RT-PCR was used to detect the expression of TMS1/ASC mRNA stimulated by medium alone and by GEM(4.27μg/ml)for 24 h and 48 h.Western blot analysis was performed with inhibitory protein of NF-κB multiclonal antibody,caspase-1 multiclonal antibody and β-actin monoclonal antibody to observe the expression of β-actin,caspase-1 and IκBα in GEM group and in the control group.The activation state of caspase-1 and NF-κB was examined.Results GEM inhibited the growth of PANC-1 cells in a concentrationand-time-dependent manners and its half maximal inhibitory concentration(IC50)was 4.27 μg/ml on 24 h.The expression of TMS1/ASC was 0.3 ±0.004 and 0.63 ±0.007 respectively in GEM group while it was 0.1 ±0.001 and 0.21 ± 0.006 in the control group on 24h and 48h.The difference between the 2 groups at the same time point had statistical significance (P ＜ 0.01).Western blot showed that GEM caused the activation of caspase-1.The expression of IκBα had no obvious differencebetween the 2 groups.GEM couldn't induce the activation of NF-κB.Conclusions GEM can inhibit proliferation of PANC-1 cells and induce their apoptosis.The drug sensitivity decreased with prolongation of exposure time.GEM might induce and increase the expression of TMS1/ASC,which might influence the apoptosis of the cells later.The apoptosis of PANC-1 cells induced by GEM is dependent of caspase-1 signaling pathway and independent of NF-κB signaling pathway.

Objective To construct adenovirus vector harboring CTLA4Ig-IRES-CTLA4 gene, and to express the gene in bone marrow mesenchymal stem cells (BMMSCs). Methods The cDNA fragments of CTLA4, and the extracellular domains of CTLA4 and IgGFc were cloned to pT-Easy vectors by RT-PCR from the activated splenocytes of Wistar rats. Recombinant adenovirus vector harboring CTLA4Ig-IRES-CTLA4 was produced from AdeasyTM Adenoviral Vector System by homologous recombination between the Adtrack vector (pAdtrack-GFP-CTLA4Ig-IRES-CTLA4) and the plasmid pAdEasy-1 containing most of the human adenovirus serotype 5 (Ad5) genome in E. coli BJ5183 strain, and then packaged and propagated in 293 cells. BMMSCs were infected with the recombinant adenovirus, and the co-expression of both CTLA4Ig and CTLA4 was then detected by RT-PCR and Western blotting. The immunosuppression function of the transfected BMMSCs was investigated by mixed lymphocyte response (MLR). Results Fusion gene CTLA4Ig was constructed successfully, and the recombinant CTLA4Ig-IRES-CTLA4 adenovirus was then generated by homologous recombination and packaged in 293 cells. The transduction efficiency of recombinant adenovirus in BMMSCs was elevated up to 98.67% determined by flow cytometry using a GFP as marker. CTLA4Ig and CTLA4 were expressed at the same time in BMMSCs detected by RT-PCR and Western blot, and the transfected stem cells showed stronger immunosuppression function than that of BMMSCs or BMMSCs transduced by Ad-CTLA4Ig. Conclusion The BMMSCs transfected by the recombinant adenovirus can co-express CTLA4Ig and CTLA4, which improved the immunosuppression function of BMMSCs in the way of costimulatory pathway blockade.

The effect of intravenous infusion of glucose-insulin-potassium (G1K) on the cardiovascular functions during acute hypoxia was studied and was compared with that of normal saline(NS). 14 anesthetized dogs were forced to inhale a hypoxic gas mixture. After the first ten-minute inhalation, Pao2 and total peripheral vascular resistance decreased to 29~3l% and 66-67% of the pre-in-halation levels respectively while the pulmonary arterial pressure increased 43-49%. Then a bolus injection of GIK was given to 8 dogs, and an injection of NS to 6 dogs. Hypoxic gas inhalation was continued for 20 more minutes. 5 -10 minutes after GIK injection, the mean arterial pressure, cardiac output, stroke volume, stroke work , and left ventricular pressure all significantly increased, however, no apparent changes could be observed in any of the above mentioned parameters after NS injection. This result reveals that cardiovascular functions during acute hypoxia can be rapidly, markedly but temporarily improved when a small volume of GIK is administered intravenously.

Animals ; Birds ; Disease Outbreaks ; prevention & control ; Hong Kong ; epidemiology ; Humans ; Influenza in Birds ; epidemiology ; prevention & control ; Poultry ; Public Health ; Public Health Administration

Objective To study the effects of subsrachnoid implantation of APA microcapsules-filled with bovine chromaffin cells (BCCs) on expression of mRNA for ?2 and ?2 subunits of GABAA receptor in spinal cord in rats with chronic constrictive injury (CCI) of sciatic nerve and to determine if GABAA receptor is involved in the mechanisms of analgesia produced by subarachnoid implantation of micro-capsulized BCCs. Methods Twenty SD rats weighing 200-250 g were randomly divided into 4 groups with 5 animals in each group : (Ⅰ) control group (group C); (Ⅱ) CCI group in which right sciatic nerve was loosely ligated; (Ⅲ) APA group in which 500-600 empty APA micro-capsules were implanted in subarachnoid space and (Ⅳ) APA-BCC group in which 5?106 APA micro-capsules filled with BCCs were implanted in subarachnoid space. In group Ⅲ and Ⅳ subarachnoid implantation was performed at L1-3 level 7 days after CCI operation. Pain threshold to mechanical stimulation with Von-Frey filament and thermal stimulation with CO2 laser was measured before and 7 days after implantation. Expression of mRNA for GABAA receptor ?2 and ?2 subunit in spinal cord was measured by RT-PCR.Results The expression of mRNA for GABAA receptor ?2 and ?2 subunit in spinal cord was significantly lower in CCI and APA groups (group Ⅱ and Ⅲ) than that in control group (group Ⅰ). In APA-BCC group (group Ⅳ) pain threshold of surgical side to mechanical and thermal stimuli and the expression of mRNA for GABAA receptor ?2 and ?2 subunit in spinal cord were significantly higher than those in group Ⅱ and Ⅲ . Conclusion The expression of mRNA for GABAA receptor?2 and ?2 subunit in spinal cord is down-regulated by CCI and subarachnoid implantation of micro-capsulized BCCs can reverse the down-regulation. Recovery of GABAA-nergic neuron activity contributes to the analgesic effect of aubarachnoid implantation of micro-capsalized BCCs.

Objective: To elucidate effects of mammalian sterile 20-like kinase 1 (MST1) gene on cell proliferation and apoptosis of human breast carcinoma cell line MCF-7. Methods: This study constructed plasmid pCMV-FLAG-MST1 and transfected it into human breast carcinoma cell line MCF-7 via mediation by Lipofectamine 2000. We detected the efficiency of transfection by Western blotting. The effect of MST1 on cell proliferation was measured by MTT assay at 12, 24, 36, and 48 h after transfection. The effect of MST1 on cell proliferation was also determined by BrdU incorporation assay at 36 h after transfection. Cisplatin was added into cultured cells at 36 h to induce apoptosis. Forteen hours later cell apoptosis was detected by Annexin V staining. Results: This study successfully constructed plasmid pCMV-FLAG-MST1. The MTT and BrdU incorporation assay showed that the cell proliferation was significantly inhibited after transfection of pCMV-FLAG-MST1. Annexin V staining demonstrated that cell apoptotic rate was relatively higher after overexpression of MST1. Conclusion: In summary, overexpression of MST1 decreases cell proliferation and induced apoptosis of human breast carcinoma cell line MCF-7.