Cytotoxic T lymphowcytes (CTL) play a major role in host anti-tumor immune responses. A major obstacle to the application of adoptive immunotherapy in the treatment of human maligancy is the inability to generate enough activated CTLs since the cytotoxic T cell undergoes activation induced apoptosis during destroying tumor cells. It is important to study how to limit activaton induced apoptosis of T cell so as to maximize the number of CTL and to enhance the tumor cytotoxicity. We have used CD3-induced Jurkat cell line as an activated T cell apoptosis model and introduced the anti-sense ICE cDNA into Jurkat T cells with retroviral vector. The effect on apoptosis of Jurkat cell induced by anti-CD3 or anti-Fas monoclonal antibody was evaluated after transfection with antisense human ICE. We found that the level of ICE expression in Jurkat cell transduced by the vector decreased and apoptosis was minimized in antisense ICE-transfected Jurkat cell after anti-CD3 or anti-Fas treatment. These results suggest that antisense blocking of ICE expression can partially inhibit Jurkat cell apoptosis induced by anti-CD3 or anti-Fas. Thus, applying antisense blocking of ICE to gene therapy may block the apoptosis of activated T cells, furthennore, enhance the antitumor effect.

Objective: To investigate the cytotoxicity of ICE gene transfection in Combination with Antitumor Chemicals killing Hepatocellular Carcinoma Cells in vitro.Methods: The recombinant plasmid pLXSN-hICE was transferred to virus packing cell PA317 by electroporation method. And then the retrovirus containing human ICE cDNA generated by these PA317 cells were used to transfect human hepatocellular carcinoma cell line HepG2. The apoptosis of transferred cells were examined by gel electrophoresis. The influence of chemotherapeutic drug Carbo-platin to the proliferation of hepatocellular carcinoma cell line SMMC7721 and its derivative cells(SMMC7721-hICE,SMMC7721-antisence hICE,SMMC7721-neo)was observed by incorporation of 3H-TDR. Results: Electrophoresis of DNA displayed the apoptosis ladder of HepG2 transfected by ICE gene. The proliferation of SMMC7721-hICE was significantly suppressed in vitro induced by Carbo-alatin compared to the other three cell lines. Conclusion: ICE gene transfection could greatly increase the susceptibility of SMMC7721 cells to apoptotic cell death following chemotherapy. These findings suggest that combining ICE gene transfection with utilizing antitumor drugs would represent a novel approach for the effective treatment of hepatocellular carcinoma.

Objective To investigate the effect of running exercise on cartilage in rats with an unstable knee joint.Methods Twenty 8-week-old Sprague-Dawley rats had their left anterior cruciate ligament cut to model an unstable knee.They were randomly divided into a control group and an experimental group,10 rats in each group.The control group was given no intervention,while the experimental group accepted running exercise training on na animal treadmill at a velocity of 15 m/min for an hour every day.After 3 and 6 weeks of training,5 rats were sacrificed and cartilage from the medial condyle of the femur was sampled,decalcificated,embedded and sliced on the sagittal plane.After hematoxylin-eosin staining,toluidine blue staining and immunohistochemical staining,the cartilage thickness,Mankin' score,the content of matrix collagen and the proteoglycan content of the cartilage matrix were assessed,and the shape and structure of the unstable knee joints were observed under a transmission electron microscope.Results The cartilage thicknesses and Mankin's scores at 6 weeks were significantly different from those at 3 weeks in both groups.In the experimental group the average thickness of cartilage was 154 ± 13 μm at 3 weeks and 131 ± 15 μm at 6 weeks.The corresponding Mankin's scores were 9.93 ± 1.36 and 11.23 ± 1.57,respectively.Both were significantly different from the control group averages at the same time points.There was also a significant difference in the positive rate of toluidine blue and collagen type Ⅱ staining between the experimental group and the control group at both time points,and in the experimental group between 3 and 6 weeks of training.After 3 weeks of training,fewer chondrocytes were observed under the transmission electron microscope in the experimental group,and fissures were seen on the surface of the cartilages.However,3 weeks later,quite a few ruptures and a lot of necrotic cells could be seen.Conclusions Running exercise can damage the cartilage of unstable knee joints and speed up the development of osteoarthritis.Even moderate exercise could aggravate damage to unstable joints and the cartilage matrix,and accelerate chondrocyte degeneration.

Objective:To investigate the effect of melatonin (MEL) influence on lipopolysaccharide (LPS)-induced long-term anxiety-like behavior and activation of astrocytes in septic neonatal rats.Methods:Sprague-Dawley rats were randomly(random number) assigned to the control group, LPS group and LPS+MEL group. Sepsis model was intraperitoneally injected with LPS (1 mg/kg), and neonatal rats in the MEL group were administered with MEL (10 mg/kg) 30 min after LPS injection. At different time points after injection, rats in each group were divided into three subgroups: 3 d, 7 d and 28 d. The expression of GFAP and TNF-α in the corpus callosum was detected by immunofluorescence staining and Western blot. Open-field test was applied to observe anxiety-like behaviors. In vitro, cultured neonatal SD rat astrocytes were divided into the control group, LPS group, LPS+MEL group, and LPS+MEL+luzindole group. Immunofluorescence staining was used to observe the expression of GFAP and TNF-α. Expression of GFAP, TNF-α, p-NF-κBp65, NF-κBp65 protein in astrocytes were assessed by Western blot. RT-qPCR was used to investigate the mRNA expression of GDNF and BDNF. One-way ANOVA and two-way ANOVA were used for comparison of multiple groups of variables. A P<0.05 was considered statistically significant. Results:LPS reduced the duration of movement in the central area and distance in the central area/total distance in open-field test, while melatonin evidently reversed the LPS-induced anxiety-like behavior. Compared with the LPS group, the expressions of GFAP and TNF-α were significantly decreased in the corpus callosum at 3 d and 7 d in the MEL group ( P< 0.05). Compared with the LPS group, MEL could significantly decrease the expression of GFAP, TNF-α and p-NF-κBp65 in astrocytes ( P< 0.05), which could be blocked by Luzindole. In addition, compared with the LPS group, MEL pretreatment could reverse the down regulation of GDNF and BDNF induced by LPS ( P<0.05). Conclusions:MEL can relieve LPS-induced long-term anxiety-like behavior in septic neonatal rats. The mechanism may be related to the inhibition of astrocyte activation and inflammatory reaction through NF - κ B pathway.

Objective Investigating and analyzing problems and influencing factors in the process of hospital-level research project management,proposing effective and feasible recommendations,and to prove a reference for hospitals administrative management development.Methods The research analyzed factors including researchers,the departments and implementation.Results Since 2005,the total funding to support research projects in the hospital-level project reached 6.345 million yuan,the pass rate in middle-term examination was 84.94％ and was 71.81％ in final examination,with the majority of them were senior researchers.Conclusions Problems founded in this study includes the participate rate was low inyoung and middle-aged researchers,the quality of hospital-level research is also low,lack of research time,and the research outcome is not satisfied.We suggest strengthen the management of the research process,improve the mechanism of scientific rewards and punishments,integrate the disciplines resources,and improve the quality of hospital-level research project.

Objective:To explore the adjustment factors to the migration and functional activity on macrophages in the uterine of inflammatory mice induced by LPS.Methods:150 Kunming female mouse were divided into control group (group A),LPS model group (group B),MCP-1 blocking-up group (group C),the mice uterines were extracted separately at hour 1,3,6,12,24.The number of CD14+ macrophages and the expression of CD14 macrophages were detected by Immunohistochemistry,ELISA detects the expression of TNF-α and MCP-1.Results:①Compared with group A,the number of CD14+ macrophages and the expression of CD14 in endometrium,myometrium,perimrtrium of group B were highly significantly increased (P<0.01) at every time points,the endometrium,myometrium of group C were closely to normal level at 1,3,6 h;compared with group B,the number of CD14+ macrophages and the expression of CD14 were highly significantly decreased(P<0.01)at 1,3,6 h of perimrtrium of group C and every time points of endometrium,myometrium of group C.②Compared with group A,the content of TNF-α and MCP-1 were highly significantly increased (P<0.01) at every time points of group B and 12,24 h of group C;compared with group B,the content of TNF-α and MCP-1 of group C were highly significantly decreased(P<0.01) at every time points.Conclusion:The migration of macrophages and the expression of CD14 and TNF-α in the uterine of inflammatory mice induced by LPS were regulated by MCP-1.

Objective To study the protein expressions of Fas-associated death domain protein (FADD) and caspase-8 in rats with intracerebral hemorrhage ,and the effects of erythropoietin tp reveal the mechanism of neu-m-protection by EPO. Methods 126 male SD rats were randomly divided into three groups: Sham-operated group, intracerebral hemorrhage group, and EPO group. Each group was divided into seven subgroups according to the differ-ent time points (3,6,12,24,48,72 h and 7 d). The model of intracerebral hemorrage was established in rats by in-tracerebral injection of autogenous blood. The protein expressions of FADD and caspas-8 in rats tissue around the hemorrhagic and the normal brain tissue were detected by immunohistochemistry. Results The protein expressions of FADD and caspase-8 were increased [(4.66±0.46 ) and ( 15.89±1.81)] at 3 h after intracerebral hemorrhage, and peaked at 48 h [ (35.88±4.24 ) and (45.04±3.99)], the expressions of FADD and caspas-8 in the region around hematoma in EPO group significantly decreased compared with model group[ (3.92±0.64) and (28.24±1.90), (13.32±2.01 ) and (35.08±2.82)] at 3 h and 48 h. Conclusion The protein expressions of FADD and easpase-8 are markedly increased after intracerebral hemorrhage. EPO can protect the neurons by signifi-cantly reducing the expressions of FADD and caspase-8.

This study is to examine the effects of NNIspm-mediated cellular senescence of HepG2 cells and elucidate its potential molecular mechanism. Cellular senescence was detected with senescence-associated beta-galactosidase staining. Cell cycle distribution, intracellular fluorescence intensity and accumulation of intracellular reactive oxygen species (ROS) were detected by high content screening (HCS). Protein expression was detected by Western blotting. Polyamines content was analyzed by high performance liquid chromatography (HPLC). The results demonstrated that NNIspm significantly induced HepG2 cells senescence. This effect was due to the decrease of intracellular polyamines, the arrest at G0/G1 phase and an increase of ROS level. The molecular senescence marker p21 increased significantly after NNIspm treatment. In contrast, the protein expressions of Cyclin E and CDK2 were obvious down-regulation. The results indicated that cellular senescence induced by NNIspm was one of its antitumor mechanisms.

might be related to the growth and metastasis of HCC.

Objective To observe the effects of acupuncture on the sleeping quality of male Wistar rats with stress induced gastric mucosal injury from the viewpoint of the brain-intestine axis. Methods Male Wistar rats were randomized into normal control, model, ST36-CV12, BL62-KI6, and combined groups, with 8 rats in each group. The model was established by using intragastric perfusion of ethanol in the rats. Acupoints of Zusanli (ST36) and Zhongwan (CV12) on both sides were used for the ST36-CV12 group. Acupoints of Shenmai (BL62) and Zhaohai (KI6) on both sides were used for the BL62-KI6 group. All four acupoints were used for the combined group. 0.22 mm × 13 mm stainless steel filiform needles were inserted 5-10 mm deep, and were left untouched for 20 minutes before withdrawal. The acupoints were stimulated each day for five days and the paw withdraw thermal latency (PWTL) was detected after the last stimulation. 2%pentobarbital sodium was then injected (40 mg/kg), and sleeping time of rats was detected. Gastric mucosa was observed through naked eyes. Gastric mucosal ulcer Index was determined by using the Guth method. 5-HT and DA contents were detected by using HPLC-ECD method. Results Compared with the normal control group, PWTL and sleeping time decreased (P<0.01), and UI, 5-HT, and DA content increased in model group (P<0.05, P<0.01). Compared with the model group, PTWL and sleeping time increased (P<0.05, P<0.01), and UI decreased in all treatment groups (P<0.01). 5-HT content in ST36-CV12 and combined groups decreased significantly, and DA content in ST36-CV12 group dropped (P<0.01). Compared with other treatment groups, PWTL and sleeping time increased significantly (P<0.01), and UI decreased obviously in combined group (P<0.01). Conclusion Stimulating acupoints of Zusanli, Zhongwan, Shenmai, and Zhaohai (KI6) may promote the recovery of gastric mucosal injury while at the same time increase sleeping time, with four acupoints used in combination having the best efficacy. The mechanism may be related to adjusting contents of 5-HT and DA in corpus striatum.