Central nervous system (CNS) injury, including brain injury and spinal cord injury, has higher disability and mortality. Therefore, CNS injury repair has been a research emphasis and focus in the field of neuroscience. The limited neurons intrinsic regenerative capacity in adult mammalians is one of the reasons of regeneration difficulties after CNS injury. However, the more important reason is the formation of inhibitive glial microenvironment at the local lesion sites. This article reviews the roles of all types of cells, such as astrocyte, microglia, taxi oligodendroglia in gtial microenvironment in CNS injury repair.

To study the chemical constituents of the fruits of Melia toosendan, three limonoids were isolated and purified by repeated silica gel column chromatography and preparative HPLC from the EtOAc extract of M. toosendan. Their structures were determined by their physico-chemical properties and spectroscopic data (1D-NMR, 2D-NMR) as: 24, 25, 26, 27-tetranorapotirucalla-(apoeupha)-1alpha-tigloyloxy-3alpha, 7alpha-dihydroxyl-12alpha-acetoxyl-14, 20, 22-trien-21, 23-epoxy-6, 28-epoxy (1), nimbolinin B (2), and trichilinin D (3), separately. Compound 1 is a new compound, and compound 2 is obtained from this plant for the first time.

BACKGROUND: Blood stem cells (BSCs) are the most primitive cells in the immune system and can be differentiated into many kinds of cells. As the regulatory cells of immune response, dendritic cells (DCs) have been attracted more and more attention in the field of autoimmune diseases. Due to different resources of precursor cells, DCs have different cytokines, ideal cytokine matching,applying orders, and experimental cultured conditions. Furthermore, development, phenotypic expression, and mature degree are still different.OBJECTIVE: To investigate the influence of tumor necrosis factor- α (TNF- a ) and interleukin-4 (IL-4) on the culture system and provide improved method for inducing functional DCs in vitro, derived from cord blood CD34+ hematopoietic precursor cells (HPC).DESIGN, TIME AND SETTING: Observational study, which was performed in the Department of Microbiology and Immunology, Nanjing Medical University from March 2005 to November 2005.MATERIALS: The cord blood was collected from neonatal umbilical cord in the 81 Hospital of Nanjing City. CD34 monoclonal antibody coated magnetic bead system (MACS) was provided by Miltenyi Biotec Company, Germany; human recombinant granulocyte-macrophage colony-stimulating factor (rhGM-CSF), human recombinant interleukin-4 (rhlL-4), and human recombinant tumor necrosis factor-α (rhTNF-α) by Pepro Tech Company, USA.METHODS: CD34+HPC were isolated and purified from umbilical cord blood by using a high-gradient magnetic cell sorting system (MACS). Then the cells were cultured with different culture medium which contained different combinations of cytokines:GM-CSF and TNF-α (GT) or GM-CSF, TNF-α and IL-α (GTI) in order to be induced to differentiate into dendritic cells (DCs).The DCs derived from CD34+HPC were identified for their morphology and phcnotype by FACS and laser scanning con_focal fluorescence microscopy, and also for their abilities of inducing proliferation of allogenic T cells by 3H-TdR incorporation assay.RESULTS: The purity of selected CD34+ cells with MACS was more than 90%. DCs could be obtained from CD34+HPC by the culture in presence of GM-CSF and TNF-α or GM-CSF, TNF- o and IL-4. With the time of culture lasting, the cells expressed lower level surface antigen of CD34 and HLA-DR (P ＜ 0.05), and possessed the phenotypes of DCs characterized by higher expression of CD80, CD86, CD83 and CDIa. At 13-15 days, the cells possessed higher level of phenotypes of DCs compared to 7-9 days and 10-12 days. DCs induced with GTI culture system expressed higher levels of surface antigen CD80, CD86, CD83,CD 1 a and a lower level of CD 14 than those induced with GT culture system. DCs displayed typical morphology and property and expressed higher level surface antigen of CD86 and CD80, especially expressed CD86 when they were induced with proper cytokines of GM-CSF and TNF- α added at 0 hour, and of IL-4 added at 48 hours.CONCLUSION: DCs can be generated from CD34+HPC by proper culture system, the design of GM-CSF + TNF- α + IL-4(GM-CSF and TNF- α were added at 0 hour, IL-4 was added at 48 hours) is preferred.

Objective To research a new type of gastrointestinal anastomat -- anastomosis device with three rows of staples (Pa-tent No .2012200583213) ,and clarify its associated functions through esophagogastric anastomosis operations on pigs ,so that we could provide experiences for its clinical application in future .Methods Compared with domestic anastomat with two rows of sta-ples ,we designed and manufactured a new type of anastomat with three rows of staples and researched its function .Choosing 12 do-mestic pigs ,about 60 kg ,gastroesophageal anastomosis was taken twice with each case by anastomat with three or two rows of sta -ples randomly in sequence .According to the different types of anastomats ,cases were divided into two groups :group A ,used anas-tomat with three rows of staples ,including 12 cases of anastomosis ;group B ,used anastomat with two rows of staples ,including an-other 12 cases of anastomosis .Results Compared with group B ,cases of group A have less bleeding sites (t = 7 .00 ,P < 0 .01) . Without reinforcement and with 0 .5 kg of tension ,fewer of outermost staples exposed(t= 6 .17 ,P< 0 .01) .And the shape of used staples of group A is double circles ,which has bigger mechanical strength than that of group B (t= 6 .57 ,P < 0 .01) .Conclusion The function of anastomat with three rows of staples surpasses that of traditional anastomat with two rows of staples in pig esopha -gogastrostomy surgery .

Objective To investigate the effect and mechanisms of factor fibroblast growth factor inducible 14(Fn14)in the high glucose induced-cardiomyocyte hypertrophy. Method To observe the expression of collagenⅠ, connective tissue growth factor ( CTGF ) , transforming growth factor-β1 ( TGF-β1 ) , and Fn14 in high glucose induced-cardiomyocyte hypertrophy. Fn14 expressions was down-regulated by siRNA interference technique, and then the expressions of collagen Ⅰ, CTGF, and TGF-β1 were observed, and the mechanism was also explored. Results The expression of collagen I, CTGF and TGF-β1 was significantly up-regulated after high glucose induced-cardiomyocyte hypertrophy for 72 h. At the same time, the expression of Fn14 was increased after 72 h-treatment, and reached the peak at concentration of 30 mmol/L high glucose. High glucose could not up-regulated the expression of collagenⅠ, CTGF, and TGF-β1 after siFn14 interference, while the same result was observed in the expression of p-JNK. Conclusion The expressions of collagenⅠ, CTGF, TGF-β1, and Fn14 in cardiomyocyte of neonatal rats were induced by high glucose. While Fn14 expression was inhibited, the expressions of collagenⅠ, CTGF, and TGF-β1 were down-regulated, which seems to be involved with p-JNK signaling pathway.

Aim To explore the effects of propofol on learning and memorizing ability and the effects of com-bination of propofol and phenobarbital sodium on epi-leptic rats.Methods Thirty-six epileptic rats were di-vided into epileptic model group (EP),normal saline group (NS),lipid emulsion +epileptic group (LE), phenobarbital sodium +epileptic group (PB),propofol+epileptic pattern (Prof),and combination of propo-fol and phenobarbital sodium +epileptic group.Each group had 6 rats.Tests of Morris water maze were giv-en to the rats to evaluate their learning and memorizing ability.The protein expression of toll-like receptor 4 (TLR4 )was examined by ELISA.Results There were no effects of saline and lipid emulsion on learning and memorizing ability and the expression of TLR4 pro-tein in hippocampus in epileptic rats (P >0.05 ). Propofol could increase the incubation period in epilep-tic rats obviously,shorten the plateau period,and in-crease the expression of TLR4 protein in hippocampus (P <0.05 ).Phenobarbital sodium could shorten the plateau period in epileptic rats,and increase the ex-pression of TLR4 protein in hippocampus (P <0.05), but it had no effect on the incubation period (P >0.05).Compared with PB,combination of propofol and phenobarbital sodium +epileptic group had a lon-ger incubation period and a shorter plateau period with an increase of the expression of TLR4 protein in hippo-campus (P <0.05 ).Compared with propofol group, combination of propofol and phenobarbital sodium +ep-ileptic group had a shorter plateau period (P <0.05) with an obvious increase in the expression of TLR4 pro-tein in hippocampus (P <0.05),but it had no effect on incubation period (P >0.05 ).Conclusions Propofol damages the learning and memorizing ability of epileptic rats.Phenobarbital sodium had no obvious effect on the learning ability in epileptic rats,but harms the memorizing ability in epileptic rats.Combi-nation of propofol and phenobarbital sodium affects the learning and memorizing ability of epileptic rats.Hip-pocampus TLR4 may be involved in the effect of propo-fol and phenobarbital sodium on the learning and mem-orizing ability of epileptic rats.

Objective To construct the lentiviral vector of RNA interference(RNAi) for DEK,and to detect its effect on breast cancer cell growth.Methods The DEK siRNA was designed and constructed based on DEK sequence using a lentiviral vector.The lentivirul vector containing DEK siRNA was named PSIH-H1-DEK as confirmed by PCR and sequenceing.PSIH-H1-DEK was then packaged with accessory plasmids into lentivirus in 293T cells and selected for 2 weeks with puromycin ( puro ) before the mixed colonies stably expressing DEK siRNA were obtained and the DEK expression was detected by real time PCR( RT-PCR) and Western blotting.The effect of DEK siRNA on ZR75-1 cell growth was determined by cell counting kit.Results Western blot and RT-PCR showed that PSIH-H1-DEK siRNA could suppress DEK gene expression.Suppression of DEK could markedly inhibit the growth of ZR75-1 cells.Conclusion The lentivirus-mediated DEK siRNA is obtained,which will facilitate further research on DEK function in breast cancer development.

Objective To explore the effects of nerve growth factor (NGF) and low frequency electricity stimulus on the configurations of skeletal muscle cells and the change of muscle fibre types in the denervated skeletal muscle separately.Methods The denervated rat model was established and model animals were injected with the NGF and given the stimulus (frequency=2 Hz) about 30 days. The configurations and the change of muscle fibre types were observed by immunohistochemistry and image analysis.Results The muscle fibre was in chaos and the boundary was not obvious among cells in the denervated rats; the muscle fibre of the denervated rats with NGF injection and low frequency electricity stimulus was more regular and the boundary of cells was clearer, the cells number was more than those of the denervated rats. Compared to normal rats, the proportion of Ⅰ muscle fibre in the denervated rats increased ( P<0.05), whereas the proportion of Ⅱ muscle fibre decreased ( P<0.05); it had no significant differences of the two types of muscle fibre between the denervated rats with NGF injection, low frequency electricity stimulus and the denervated rats ( P<0.05).Conclusion NGF injection and low frequency electricity stimulus can make the configurations of denervated muscle to better.

Objective To construct the lentiviral vector (pSIH-H1) for E6AP small-interfering RNA(siRNA) and to detect its effect on breast cancer ZR 75-1cell growth.Methods E6AP siRNA was designed and constructed based on hu-man papillomavirus E6-associated protein ( E6AP) cDNA sequence.The expression of E6AP was examined by real-time quantitative PCR(qRT-PCR) and Western blotting.The effect of E6AP on ZR75-1 cell growth was determined by cck-8 kit.Results DNA sequencing indicated that E 6AP siRNA expression vector was constructed successfully .qRT-PCR and Western blotting experiments showed that pSIH-H1-E6AP siRNA could suppress the E6AP gene expression.Suppression of E6AP could markedly inhibit the growth of ZR 75-1.Conclusion A lentivirus RNA interference ( RNAi) vector targeting E6AP gene is successfully constructed ,which inhibits the cell growth of ZR 75-1.

Objective To detect the effect of E6AP on gastric cancer cell proliferation and migration.Methods The expression of E6AP in different gastric cancer cell lines and normal gastric mucosa epithelial cell lines was detected by Western blotting.Gastric cancer cells BGC-823 stably expressing E6AP short hairpan RNA(shRNA) were obtained by lentiviral vector of E6AP.The effect of E6AP on BGC-823 cell growth and migration was determined by CCK-8 kit, Tran-swell and wound healing assay.Results Gastric cancer cell line BGC-823 in which E6AP was stably knocked down was established.Knockdown of E6AP inhibited the proliferation and migration of BGC-823 cells.Conclusion E6AP plays a key role in gastric cancer proliferation and migration.