AIM:To study the effects of lipopolysaccharide(LPS), interleukin-6(IL-6)and tumor necrosis factor ? (TNF?) on tissue factor(TF) expression of astrocytes. METHODS:Astrocytes were identified with anti-glial fibrillary acidic protein antibody. The TF activity of cell lysate was measured with one stage clotting assay. RESULTS:TF activity of astrocytes of LPS,IL-6,TNF? groups were obviously higher than that of the control group( P

To exert the role of high quality resources of national experimental teaching demonstration center in teaching,we have promoted laboratory opening through research teaching.By the contest of university students’experiment designing and inviting public bidding for exploring experiment,we encourage students to conduct innovation research,blazed their orexis and enthusiasm to start scientific research.We also encourage teachers to explore new teaching methods and innovated experimental items.

Objective To analyze the relationship between needs of assistive devices and the three other factors (age, disability categories, severity levels of disability) in order to scientifically formulate subsidy policy on assistive devices.MethodsThe assessment of rehabilitation needs for 928 Shenzhen citizens with new disability certificates in its six districts was performed by professionals, and the data was recorded into ACCESS database and analyzed.ResultsThe sequence of demand rates on assistive devices among disability categories was vision disability (78%), hearing disability (76%) and physical disability (74%); but speech, mental retardation and psychological disabilities had a low demand. The demand rates of assistive devices were dissimilar among four age groups for persons with vision, hearing and physical disabilities; for severity levels of three disability categories of vision, hearing and physical disabilities: the milder the vision disability was, the greater demand rates of assistive devices were; the demand rate of those with the second level of hearing disability was the highest; the demand rates of persons with the first and second levels of physical disability were relatively higher.ConclusionWhen drawing up subsidy policy on assistive devices for persons with disability, the disability categories, ages of persons with disability and severity levels of disability should be considered; the different types of assistive devices should be supplied according to rehabilitation goals at different age group; and necessary assistive devices should be provided according to severity levels of disability.

Objective:To investigate the dynamic changes of the luminal microbiota in the jejunum following administration of proton pump inhibitors (PPIs) in a rat model.Methods:Rats were randomized into six groups (n =6 each group).A group of rats were sacrificed just after anesthesia as normal control (0 d) and,other five groups were continuously administered with omeprazole (10 mg/kg twice daily,intraperitoneally) and were euthanized at 5,9,14,21,28 days following the treatment,respectively.Total DNA in the luminal contents of jejunum was extracted and was used for polymerase chain reaction (PCR) amplification with the primer set targeted the hypervariable V3 region of 16S ribosomal RNA genes.Subsequently,the amplicons were separated by denaturing gradient gel electrophoresis (DGGE).After the gels were stained and photographed,the bands were cut out and sequenced to determine the closest bacterial relatives with the BLAST.The DGGE profiles were analyzed to evaluate the shifts of the microbiota composition and diversity following treatments.Results:Changes of the jejunal microbiotas in rats were observed at 5 and 9 days post PPI administration,as characterized by outgrowth of Streptococcus pneumonia,Clostridium saccharolyticum and Lactococcus garvieae compared to those of the controls (0 d).With time extension of PPI treatment,the mictobiotas significantly shifted toward dysbiotic state,in which the opportunistic pathogens,including Ertterococcus faecalis and Clostridium difficile,were strikingly expanded,especially 21 days later.However,the commensals such as Lactobacillus reuteri and Weissella koreensis were markedly declined in PPI-treated animals compared with the controls.The similarity of the jejunal microbiotas between PPI-treated animals and controls was markedly reduced following PPI treatment,reaching (56.1 ± 16.7) ％ at 28 days.Conclusion:Our data demonstrate that the gastric acid suppression could induce shifts of the jejuna microbiota in a rat model.More importantly,long-term use (＞ 14 d) of PPI could lead to the dysbiosis of the jejunal microbiota,which might be related causally to increased susceptibility to enteric infection.

Objective To investigate the change of dynamic expression of t-PA and PAI-1 during early venous crisis after free perforator flap transplantation.Methods Thirty healthy New Zealand white rabbits weighed 2.5-3.0 kg were chosen and randomly divided into experimental group (n =15) and control group (n =15).Free transplantation of superficial epigastric artery perforator flap (SEAPF) was implemented in all rabbits firstly.Then the model of venous crisis was established by ligating the anastomosis vein in order to interrupt venous blood outflow in experimental group.The blood supply of all flaps was monitored by observing their color,swelling degree and the filling reaction of the capillaries after operation.Peripheral blood was drawn from femoral artery at different time point for measuring the concentration of t-PA and PAI-1 by Elisa.Partial flap tissue was harvested for pathological examination at corresponding time point.Data analysis was performed by using SPSS 17.0 statistical software.P ＜ 0.05 was considered statistically significant.Results One rabbit died of anesthesia,and the venous congestion was observed in 1 rabbit in control group.The models of free transplantation of SEAPF and venous crisis were established successfully in the remaining rabbits.No significant appearance change was observed within 1 h after the outflow vein being ligated,while typical appearance of venous crisis could be observed 2 hours after the outflow vein being ligated.Compared with the control group,the concentration of t-PA was lower,but the concentration of PAI-1 was higher in experimental group at 2 hours,4 hours,6 hours,8 hours after the outflow vein being ligated(P ＜ 0.05).However,there was no obvious differences between two groups at other time points (P ＞ 0.05).The pathological examination showed the red cells gradually got together and adhered to the venous wall,eventually the microcirculation had been blocked completely and theflap became necrosis after venous crisis being occurred.Conclusion t-PA and PAI-1 can't be used to diagnose early venous crisis of perforator flap transplantation.

Objective To investigate the resistance mechanisms and epidemiology of Enterobacteriaceae isolated from clinical with reduced susceptibility to imipenem or meropenem.Methods 18 strains of Enterobacteriaceae with reduced carbapenem susceptibility were collected during January to August in 2010.The MICs of these strains were determined using automated microbial identification system.ESBLs,AmpC and KPC were tested using the agar dilution method.PCR amplification and DNA sequence were performed to analyze the KPC genes,PFGE was used to examine the molecular epidemiology.Results All 18 strains were detected ESBLs and AmpC,14 strains were detected KPC-2.3 strains with EDTA paper method positive may produce other metal carbapenem,in which 2 strains harbor KPC-2.PFGE types indicate that there were six genotypes among 15 strains of Klebsiella pneumoniae.Conclusion Plasmid-mediated KPC-2 was the main reason which makes Enterobacteriaceae reducing carbapenem susceptibility and causes short-term epidemic in hospital.Clinical strains harboring KPC-2 gene may carry multiple resistance genes meanwhile.

AIM: To evaluate the implication of CXCL10-loop3-EGF fusion protein for the activities of targeting tumor and anti-angiopoiesis. METHODS: RT-PCR was preformed to amplify CXCL10 coding sequence from PBMC activated by IFN-γ. CXCL10-loop3-EGF fusion gene, which was conducted by Over-Lap Extention PCR, was hinged up with plasmid pTG19-T, transfected to E. coli DH5α and processed positive colony selection. After ligated with plasmid pET32a(+), recombinant CXCL10-loop3-EGF fusion gene was then transfected to E. coli Origami B (DE3) and induced to express its coding fusion protein his-CXCL10-loop3-EGF. The recombinant fusion protein CXCL10-loop3 -EGF was purified by His-bind affinity chromatograph, enterokinase cleavage, ultrafiltration and dislysis. The transwell chemotatic test and HUVEC angiopoiesis inhibition test were performed to determine the anti-tumor responses and anti-angiopoiesis activity of CXCL10-loop3-EGF fusion protein. RESULTS: CXCL10-loop3-EGF fusion protein was successfully constructed and confirmed by SDS-PAGE analysis and Western blotting. Significant PBMC chematatic activity and HUVEC anti-angiopoiesis activity were observed. CONCLUSION: CXCL10-loop3-EGF fusion protein, which has perfect anti-tumor activity, is successfully constructed.

AIM: To investigate the immunologic mechanism of CXC chemokine ligand 10(CXCL10) and its receptor CXC chemokine receptor 3 (CXCR3 ) involved in the process of endometriosis (EM). METHODS: Serum samples were collected from 3 groups; EM patients without operation (n = 76) , EM patients with operation (n = 10) and the normal control persons (n =76). CXCL10 and CA12S concentrations were detected by means of ELISA and chemilumino-metry. Cell surface antigens on the activated PBMC - CD3 and CXCR3, as well as CXCR3 subgene - CXCR3A and CX-CR3B were tested by flow cytometry (FC) and RT - PCR when PBMC was separated from women with EM ( n = 10) and without EM (n = 10), and then activated. RESULTS: Serum CXCL10 concentrations between three groups were signifi-canly different (P < 0.05). Compared to normal control group, although the supernatant CXCL10 concentration and CD3~+ /CXCR3~+ PBMC number in EM group has no significant difference (P >0.05) , highly expressed CXCR3B in EM group rather than CXCR3A was observed. CONCLUSION: CXCL10 in women with EM is low, indicating that it plays a vital role in the process of EM and immune system of the women with EM is defected and impaired. The immunoreactivity of PBMC from both EM patients and normal person is same to activated signal, but the productions are different: PBMC in EM group mainly express CXCR3B but PBMC in normal person mainly express CXCR3A after activation, which may be one of the immune mechanisms that EM escapes from immunological lethal effect of the infected host.

Objective To analyze and summarize the experiences in diagnosis and treatment of patients with severe traumatic brain injury (sTBI) so as to increase the cure rate and survival rate. Methods A retrospective study was made on the diagnoses and treatment of 4 462 cases of sTBI inclu-ding 3 298 males (73.9%) and 1 164 females (26.1%). The most frequent cause for injury was traffic accident, accounting for 35.5% (1 583/4 462). Closed bead injury ocurred in 3 654 cases (81.9%) and open head injury in 808 (18.1%). The most commen clinical manifestations included unconscious-ness, changes of pupils and life signs. Of all, 1 158 cases (26.0%) were found with combined injury and 1 356 (30.4%) with complications. Treatment measures included first aid, surgery or conventional treatment. Emphasis was put on the treatment of secondary insults. Results Surgery was performed in 3 023 cases (67.7%) including craniotomy for hematoma clearance in 856 cases and hematoma clearance plus decompression in 2 167, with mortality rate of 17.9%. Conventional treatment was carried out in 1 439 cases (32.3%), with mortality rate of 23.7%. There were 2 462 eases (55.2%) with fair recov-ery, 508 (11.4%) with mild disability, 339 (7.6%) with severe disability, 272 (6.1%) under vege-tative state and 881 (19.7%) deaths on discharge according to the Glasgow Outcome Scale. Conelu-sions Active diagnoses and treatment, strict medication rules and prevention and treatment of secondary insults may be key to higher eure rate as well as lower morbidity and mortality of sTBI.

Objective To analyze the relationship between 1082 site A/G polymorphism in interleukin-10 gene and different virulence factors of Helieobacter pylori in Gansu Province and susceptibility to gastric cancer genesis. Methods Polymerase chain restriction fragment length polymorphism and direct sequencing were performed to analyze the genotype of the A/G polymorphism in its-1082 site of promoter region, and immunoblotting was performed to test different virulence factors antibody of H. pylori. Results respectively in the gastric precancerous lesion group. The frequency of AG + GG genotype was statistically higher in the gastric precancerous lesion group compared to the control group (P =0. 018), the risk gastric frequency of AA, AG, GG genotypes were 58.4% ,35.8% ,5.8%, respectively in the gastric cancer group. The frequency of AG + GG genotype was higher in the gastric cancer group compared to the control group (P =0. 010), moreover, individuals with the IL-10-1082AG + GG genotype rose to 2. 31 fold risk for gastric there was an increased risk of gastric precancerous lesion and developing gastric cancer for those carrying both AG + GG genotype and seropositive H. pylori I, with 9.73 fold risk. Conclusion There was a relationship between IL-10-1082 A/G polymorphism and susceptibility to gastric cancer.