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Objective To explore the formation of intravascular microthrombosis in the brain after traumatic brain injury in rat in order to study the potentially causal linkage between thrombosis and post-traumatic cerebral ischemia. Methods Lateral fluid percussion injury (FPI) model was reproduced in the present study to prodace acute brain injury in rats. The rats were sacrificed at 12, 24, 72 hours and 7days after traumatic brain injury. Sham-injured animals received identical craniotomy without FPI. Pathological studies were performed to study the distribution and severity of intravascular microthrombosis in the brain. The difference in the severity of intracerebral thrombosis between groups was evaluated. Results Abundant fibin thrombi were found in all injured rats as compared with the controls (P

Objective To establish a simple and reliable method for primary culture and purification of embryonic dorsal root ganglion neurons. Methods Dorsal root ganglions (DRG) from rat embryos were isolated under microscope and dissociated in trypsin. The dissociated neurons were treated with 5-fluorouracil for purification. Immunocytochemistry was performed by double staining with anti-calcitonin gene-related peptide and anti-NF200 antibodies to evaluate the purification rate of DRG neurons. Results Isolated and cultured DRG neurons survived and grew nicely in the basis medium for neurons, and the purified rate of DRG neurons could reach to around 93%. Conclusion The above method is very useful to obtain dorsal root ganglion neurons and to purify the neuronal population.

Objective To explore the potential mechanism of efficacy of imipramine treatment in the regeneration of hippocampus neuron after traumatic brain injury in rat.Methods Lateral hydraulic battering injury(HBI)was used to induce acute brain injury in rats.Forty male SD rats were randomly assigned to four groups:HBI with imipramine treatment group,HBI without treatment group,sham injury with imipramine treatment group and sham injury without treatment group.Animals of all the four groups were sacrificed for histological evaluation two months after treatment.Proliferating cells and neurons in hippocampus were counted and compared between groups.Results Proliferating cells and neurons were found to be obviously increased in all injured rats compared with sham injured animals(P

AIM To explore whether plasma vasoactive substances and neutrophil infiltration make different contribution in cerebral ischemia-reperfusion injury and protective effect of triacetylshikimic acid(TSA). METHODSThe rat models of ischemia 90 min and reperfusion 3-48 h were prepared with middle cerebral artery occlusion. TSA 50-200 mg·kg-1 were given (ig) immediately and 60 min again after the onset of ische- mia. Serotonin and thromboxane B2 (TXB2) concentrations in plasma were detected by fluorescence spectrophotometry and radioimmunoassay respectively. Myeloperoxidase(MPO) activity in brain tissue was quantified by chemical analysis. RESULTS At 3-24 h after reperfusion, the concentrations of plasmic serotonin, TXB2, and brain MPO activity increased obviously in a time-dependent manner. At 48 h after reperfusion, the concentrations of serotonin and TXB2 decreased to the same level of sham. Nevertheless, brain MPO activity remained more elevated than the contralateral cortex. At 24 h after reperfusion, TSA (100 and 200 mg·kg-1) was shown to possess the ability to inhibit the increased plasmic serotonin, TXB2 concentrations, and brain MPO activity induced by focal cerebral ischemia-reperfusion. CONCLUSION Vasoactive substances in plasma and MPO activity in brain tissue show different time courses during focal cerebral ischemia-reperfusion and make different contribution to brain damage. TSA is effective to protect the ischemic brain tissue from ischemia-reperfusion injury.

Objective The aim of this study was to assess the value of △Np63α in predicting tumor recurrence after curative resection in esophageal squamous cell carcinoma (ESCC) patients.Methods We analyzed △Np63α protein cxpression in 304 clinicopathologically characterized ESCC cases by immunohistochemistry.Results We found △Np63α expression was positive in 122 (40％) of 304 cases.△Np63α expression was higher in the cancer tissue than in non-tumorous control tissue at protein level(P =0.034).There was a significant difference of △Np63α expression in patients categorized according to invasive depth (P =0.001),tumor position (P =0.001) and lymph nodes metastasis condition (P =0.001).Multivariate analyses showed that △Np63α was an independent prognostic marker for ESCC recurrence.Conclusion △Np63α is associated with outcome of ESCC and can be a novel predictor for poor prognosis of ESCC patients after curative resection.

Retroviral expression system which consists of retroviral vector,envelop protein vector and packaging cell line is an efficient expression system for recombinant protein.It has great potential in gene therapy and biopharmacy.Transcriptional active genome regions are the preferred targets for retrovirus integration.Furthermore,VSV-G protein enables this system a broader host range and makes virus integration more efficient.After infection of high-titer virus,high production clones can be selected through simple screening.So far,the research on retrovirus expression system has developed into application in bio-pharmacy industry.Here the composition of this system and the mechanism of virus transduction and summarize the application and prospect of retroviral expression system are introduced.

Objective To investigate the efficacy and safety of the postoperative analgesia after thoracotomy with different patient-controlled intravenous analgesia (PCIA)medication.Methods One hundred and twenty ASA Ⅰ or Ⅱ patients scheduled for elective thoracic surgery were randomly di-vided into four groups according to different PCIA formula:group S received sufentanil 3 μg/kg;group SD received sufentanil 1.5 μg/kg plus dezocine 0.3 mg/kg;group SF received sufentanil 1.5μg/kg plus flurbiprofen axetil 3 mg/kg;group DF received dezocine 0.3 mg/kg plus flurbiprofen ax-etil 3 mg/kg.For all four goups,the applied medicine was diluted with 0.9% saline to 300 ml,and was infused at 5 ml/h with a bolus dose of 3 ml.The lockout time was set at 1 5 min.The VAS and Ramsay sedation scale were recorded 2,4,8,24,48 h after surgery,respectively.Total pressing times and the side effects were also recorded for the 24 h period after the surgery.Results There were no significant differences of the VAS score at the time 2,4,8,24 and 48 h post surgery of press num-bers within 24 h postoperatively among all groups.The Ramsay sedation scale was significantly high-er at the time 2,4,8,24 h after surgery in group S than those in other three groups (P <0.05).The side effects were more in group S than those in other three groups (P <0.05).Conclusion Compared with sufentanil alone,combination therapy is safer and more effective for analgesia after thoracic sur-gery.With good outcomes and less side effects,it deserves more consideration in clinics.

Objective To explore the influence of the lentiviral vectors mediated mouse genetic engineering regulatory T cells(Tr) infused after allogeneic bone marrow transplantation(allo-BMT) on graft-versushost disease(GVHD) in mice. Methods Lentivirus-mediated expression of forkhead box P3 (Foxp3) converted CD4 + CD25 - T cells from BALB/c mice into engineered Tr in vitro. An allo-BMT model of BALB/c→C57BL/6 mice was established. After irradiation, the recipients were injected with donor cells along with genetic engineering Tr. Survival time, histopathological analysis, serum levels of inflammatory cytokines were observed after allo-BMT. Results The mean survival times in radiation group, transplantation control group, engineering Tr group and empty vector control group were ( 8.8 ± 0.6 ) d, ( 36.7 ± 2.5 ) d, ( 51.6 ± 4.0 ) d and ( 34.1 ± 2. 3 ) d. The survival time in engineering Tr group was significantly increased as compared to other groups as judged by the log-rank test ( P ＜0.05 ). Histopathological analysis in several target organs( skin, liver and small intestine) confirmed the presence of severe GVHD in transplantation control group and empty vector control group. No histological signs of GVHD were observed in recipients in engineering Tr group. The serum levels of IFN-γ, IL-2 and TNF-α were all increased after transplantation in above groups. The peaks of concentrations of IFN-γ, IL-2 and TNF-α in engineering Tr group were significantly decreased compared to transplantation control group and empty vector control group at day 21 ( P ＜ 0. 05 ). Conclusion Co-injection of genetic engineering Tr can efficiently prevent recipients from lethal GVHD during allo-BMT in mice by reducing the serum levels of inflammatory cytokines.

Objective To explore the influence of the lentiviral vectors-mediated mouse genetic engineering regulatory T cells (Treg) infused after allogeneic bone marrow transplantation (alloBMT) on graft-versus-host disease (GVHD) and graft-versus-leukemia (GVL) effect in mice.Methods Lentivirus-mediated expression of Forkhead box P3 (Foxp3) transformed CD4 + CD25- T cells from Balb/c mice into engineered Tregs in vitro. An allo-BMT model of Balb/c→C57BL/6 mice was established. The recipients were given lethal X-ray total body irradiation before transplantation.Mice were randomly assigned into five groups and each group contained 10 recipients: (1) The recipients in radiation group were injected with 0.2 ml RPMI 1640; (2) The recipients in leukemia control group were injected with 5 × 106 donor bone marrow cells and 500 mouse T-cell leukemia/lymphoma cells (EL4 cells); (3) The recipients in transplantation control group were injected with 5 × 106 donor bone marrow cells and 5 × 106 splenocytes plus 500 EL4 cells; (4) The recipients in engineering Treg group were injected with 5 × 106 donor bone marrow cells, 5 × 106 splenocytes and 500 EL4 cells plus 5 × 106 genetic engineering Treg; (5) The recipients in empty vector control group were injected with 5 × 106 donor bone marrow cells, 5 × 106 splenocytes and 500 EL4 cells plus 5 × 106 empty vector-transduced CD4+ CD25- T cells. Survival time, clinical GVHD score or histopathological analysis (skin, liver and small intestine) were observed after allo-BMT. Chimerism of bone marrow cells from recipients survived for 60 days after transplantation was measured. Results The mean survival time in radiation group, leukemia control group, transplantation control group,engineering Treg group and empty vector control group was ( 10. 3 ± 1.5), (20. 7 ± 1.9), (26. 0 ±4.3), (49. 0 ± 17. 7) and (24. 4 ± 4. 1 ) days respectively. The survival time in engineering Treg group was significantly prolonged as compared with other groups as judged by the log-rank test (P＜0. 05).Histopathological analysis in several target organs (skin, liver and small intestine) confirmed the presence of severe GVHD in transplantation control group and empty vector control group. No histological signs of GVHD or leukemia were observed in recipients in engineering Treg group and clinical GVHD scores in this group were significantly decreased as compared with transplantation control group and empty vector control group. Conclusion Co-injection of genetic engineering Treg can efficiently prevent recipients from lethal GVHD without affecting GVL activity during allo-BMT in mice.