Objective:To sutdy the effect of 2 chloroadenosine(2 ClA),which is specifically cytotoxic to macrophages,on MTT assay in activation test and cytotoxicity test of lymphocyte.Methods:Using cell culture technique,mouse splenic lymphocytes and peritoneal macrophages were cultured.Lymphocyte activation and specific cytotoxicity to tumor cells and toxicity of 2 ClA to macrophages were measured by MTT assay in the presence or absence of 2 ClA.Results:2 ClA had a strong cytotoxic effect on macrophages.When the activation test and cytotoxicity test of lymphocyte were measured by MTT assay,the optical density values of 2 ClA group was lower than that of control group,and statistic analysis showed P

Objective:To study the specific antitumor effect of DC modified by Hsp70 tumor peptide complexes in vitro and in vivo.Methods:The tumor antigen peptides were acquired from H22 liver cancer cells and bound Hsp70 in vitro by using biochemical technique;the mouse marrow cells were cultured with induction of rmGM CSF and rmIL 4 by using cell culture technique;mouse spleen lymphocytes was stimulated.The cultured DC cells were harvested and activation of lymphocytes was detected by MTT test and cytotoxicity of stimulated and proliferated lymphocytes to H22 tumor cells and Ehrilich ascites carcinoma cells was tested;The inhibitation to tumor was observed in vivo,after stimulated DCs were injected in mice inoculated by tumor cells.Results:DCs could become mature with the effect of Hsp70 H22 peptide complexes and secret IL 12?TNF ??IL 1? and effectively activate lymphocyte;The activated and proliferated lymphocytes could specifically kill H22 cells but not Ehrilich ascites carcinoma cells in vitro;DCs modified by Hsp70 H22 peptide complexes could become one useful kind of vaccines to inhibit H22 tumor growth in vivo.Conclusion:DCs orignied from marrow cells can be effectively modified by Hsp70 H22 peptide complexes,these modified DCs can specifically activate lymphocytes in vitro and effectively induce antitumor immune response.

Objective: To investigate the effect of the dosageof chemotherapeutic agent on tumor chemotherapy linked with biotherapy and provide experimentalevidence for the dose choice of chemical drugs in the combination of chemotherapy and biotherapy.Methods: The high- and low-dosage of MMC was determined by injection of mice with different dosages of MMC. Mice inoculated with H22 hepatic carcinoma cells were treated with different dosages of MMC followed by three different kinds of biotherapy: transfection with plasmid pCH510 in vivo, immunization with Hsp70-tumor antigen peptide complexes and the combination of these two elements. Results: By toxicity test of MMC to mice, it was determined that 100 ?g of MMC was high dosage and 50 ?g was low dosage. The curative effect was significantly improved if chemotherapy was followed by different elements of biotherapy. Better efficacy was obtained when biotherapy elements were used to follow the chemotherapy with high dosage of MMC. In the case of low dosage of MMC, no difference could be observed in curative effect of three different kinds of biotherapy. When high dosage of MMC was used, the curative effect of three different kinds of biotherapy was signiferently different. The best efficacy was obtained if chemotherapy was followed by the combination of two biotherapy elements, transfection with plasmid pCH510 in vivo and immunization with Hsp70-tumor antigen peptide complexes. Conclusions: Using different chemical dosages, the curative effect of chemotherapy linked with biotherapy is different. In the case of high dose, the chemotherapy linking with biotherapy can reach more better efficacy.

Objective: To invistigate the relationship between time and efficacy of the linkage of tumor biotherapy after chemotherapy by studying the influence of chemotherapeutic drugs on immunocytes.Methods: The cytotoxicity of chemotherapeutic drugs to tumor cells, mouse peritoneal macrophages and spleen lymphocytes was observed by cell culture technique. The influence of chemotherapeutic drugs to the metabolism and activation of macrophages and lymphocytes at different time after peritoneal injection of drugs was observed. The mice were inoculated with tumor cells two days after the injection of drugs, and the growth of tumor was measured 14 days after inoculation by anatomizing mice. Results: Chemotherapeutic drugs had cytotoxicity in vitro to different cells, suppressed the function of immunocytes, and decreased the number of immunocytes in vivo. After injection of drugs, the number of immunocytes was the lowest on the third day and recovered to the nomal level on the 10th day. If drugs was injected two days before the inoculation of tumor cells, the growth of tumor became faster than control group. Conclusions: Chemotherapy not only decreases the number of immunocytes but also suppresses the function of immunocytes, and it can promote the growth of tumor after its cytotoxicity disappeared. So it is not good that biotherapy, which depends on immunocyte to kill tumor cells, is used immediately after chemotherapy and it is also not good for using biotherapy with a long interval after chemotherapy . It is good time to use biotherapy when the number of immunocytes is lowest or the recovery just starts.

Objective To study the effect of omethoate on the activity of characteristic enzymes in mouse testicles and the antagonism of teapolyphenol. Methods 80 Kunming male mice were divided into 8 groups, omethoate was given by gavage: the control group given 0.9% NaCl solution, a group given teapolyphenol of 180 mg/kg bw, three groups given omethoate of 1, 2 and 4 mg/kg bw respectively, three groups given teapolyphenol of 180 mg/kg bw and two hours later followed by omethoate of 1, 2 and 4 mg/kg bw respectively. The gavages were conducted for 6 consecutive days. The activities of the characteristic enzymes (AKP, ACP, LDH and LDH-x) in mouse testicles were measured by biochemistry method and variance analysis was done with the SPSS 11.0 software. Results As omethoate dose increased, the body weight and the testicle weight decreased obviously, the activities of AKP, ACP, LDH in mouse testicles significantly increased compared with the control. Given teapolyphenol 2 hours before given omethoate, the significant changes as above did not emerged. Conclusion Teapolyphenol has an antagonism against the adverse effects of omethoate on the activity of characteristic enzymes in mouse testicles.

BACKGROUND:Drug eluting stents and endothelium stents for clinical treatment of vascular stenosis can lead to delayed endothelialization and restenosis. A rapamycin eluting stent combined with CD34 antibody can play a synergistic role to offset delayed endothelialization and intimal hyperplasia due to antiproliferative drugs, but it is stil in the pilot phase. OBJECTIVE:To observe the ability of rapamycin eluting stent combined with CD34 antibody to capture endothelial progenitor cels, and to observe the differentiation characteristics of the captured cels. METHODS:Scanning electron microscope and indirect immunofluorescence were used to observe the morphology and differentiation characteristics of captured endothelial progenitor cels. Under a fluorescence microscope, we observed the captured endothelial progenitor cels and the degree of endothelialization after implantation of the rapamycin eluting stent combined with CD34 antibody into rabbit ear vein. RESULTS AND CONCLUSION:Under the scanning electron microscope, fusiform-like cels with a diameter of 6-8 μm were captured by the composite stent, and 24 hours later, the cels became ful-shaped. The captured cels had the appearance characteristics of endothelial progenitor cels. Results from indirect immunofluorescence observation showed that there were a lot of red fluorescent spots on the coating which represented adherent cels positive for vascular endothelial growth factor receptor-2; the composite stent was largely covered with vascular endothelial cels at 24 hours after stent implantation, and fuly covered at 48 hours, but there was no abnormal cel cluster. These findings indicate that the rapamycin eluting stent combined with CD34 antibody can be specific to rapidly capture endothelial progenitor cels in the peripheral blood, and the stent can be completely covered with vascular endothelial cels at 48 hours after stent implantation, thereby achieving rapid endothelialization and promoting the repair of endothelial cels.

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Objective　To obtain mammalian cell expression vector of human CD154 gene. Methods　A 820 bp cDNA fragment was amplified by RT-PCR method from total RNA of human peripheral blood mononuclear cell（PBMC） activated with 10 ng／mL PMA and 1 μg／mL PHA for 8 hours. The fragment was cloned into pcDNA3.1（＋） plasmids.The cloned insert was identified by double digestion of the recombinant plasmid with restriction enzymes BamH Ⅰ and EcoR Ⅰ and sequenced by Sangers-dideory-mediated chain termination. Results　This cDNA fragment included 820 bp entire coding region and a part of the 3 non-coding region. The recombinant mammalian cell expression vector of pcDNA3.1（＋）／hCD154 was constructed， the sequence of the insert was identical to the published sequence encoding human CD154 antigen. Conclusion　The recombinant mammalian cell expression vector of pcDNA3.1（＋）／hCD154 was successfully constructed.

Spinal cord injury (SCI),which lead to sensory and motor function impairment,refers to traumatic injury to the spinal cord.The main pathophysiological changing process can be divided into two phases:the preliminary injury phase and the secondary injury phase.Organ dysfunction,tissue necrosis,sensory and motor function of irreversible lesions and even death can be led by SCI and the secondary injury phase.In order to prevent the secondary injury and repair the injured central nerve system,medication,surgery,biomaterials transplantation and autologous peripheral nerve transplantation have been widely studied.However,these strategies can only reach a certain level of symptoms relieve and neural restoration,which cannot reach clinical satisfaction.Therefore,SCI has been treated as a global difficult and attractive topic.Since stem cells have the potential ability to promote neurons regeneration and/or even differentiate into neural like cells,they may also have anti-inflammatory,anti-apoptotic,anti-oxidative stress,and promote secretion of trophic factors and vascularization,they brought new hope for SCI patients.Mesenchymal stem cells (MSCs) have attracted many researchers' attention for their multi characteristics including widely distribution,strong differentiation potential,easily isolating and storing and ethics avoiding.It has been proved that MSCs are effective for repairing SCI in vivo and vitro.In the latest 3 years,with the consummation of several clinical trials,a good prospect have been showed in the translation of MSCs transplantation into clinic.MSCs have been proved to have a significant prospect for clinical translation.At the same time,more specific issues were also raised.Thus,based on the latest 3 years clinical research and the author team's latest experimental results,we reviewed the MSCs on treating SCI from basic science to clinical trial and discussed its future development.

Objective To investigate the significance of miR-193b to biological behaviors of hepatocellular carcinoma (HCC). Methods 48 cases of HCC specimens and corresponding adjacent tissues were collected, and the miR-193b expression levels in these specimens were measured by real-time quantitative PCR. The miR-193b expression was measured by the same way in HepG2 and SMMC-7721 cells. The HepG2 and SMMC-7721 were transfected with miR-193b mimics or negative control miRNA mimic with Lipofectamine 2000, and the non-transfected cells were taken as blank control. The proliferation ability of the HCC cells were detected by MTT method, and the apoptosis rate was tested by flow cytometry. Results The expression level of miR-193b in HCC tissues (2.441 ±0.569) was significantly lower than that in the corresponding adjacent tissues (15.488±4.326) (P < 0.05). Compared with normal liver cell line L-O2, the expression levels of miR-193b were significantly lower in HepG2 and SMMC-7721 cells. Transfected with miR-193b mimic, the proliferation ability of HepG2 and SMMC-7721 cells were reduced, while their apoptosis were increased. Conclusion miR-193b may be negative to regulate the proliferation of HCC and increase its apoptosis.