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 evaluate the long-term efficacy and safety profile of alpha-1-thymosin (T?_1) combined with Lamivudine(LAM) in the patients with chronic hepatitis B. Methods Eighty patients with chronic hepatitis B were randomly assigned by 1∶1 proportion to be given 100 mg LAM orally alone (LAM group) or T?_1 1.6 mg subcutaneous injection, combined with LAM(LAM+T?_1 group). Results 51.4 percent (18/35) of the patients achieved HBeAg seroconversion in combination group, while 5.4%(2/37) of the patients in LAM group achieved HBeAg seroconversion at 52 week, P

Objective To study the relationship of hepatitis and pregnan cy .Methods 49 cases with viral hepatitis of pregnancy were analyze d and compared with viral hepatitis of non-pregnant cases.Results Most of the hepatitis following pregnancy were HBV,with the increment of preg nant weeks,the incidence and mortality rates of hepatitis were increased.PT was remarkably prolonged and ALB lowered in 49 cases.Complications of hepatic enceph alopathy and hepto-renal syndrome were more,the incidence and fatality rates of severe hepatitis were higher than that in healthy group (P

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 investigate the expression of 4-hydroxynonenal (4-HNE) in the kidney of diabetic rats and the effect of probucol.Methods The rats were being intraperitoneal injected with STZ (60 mg/kg) to establish diabetic models.Then diabetic rats were randomly divided into diabetic group (group D,n =24),probucol treated group (group P,n =24).Normal rats were taken as control group (group C,n =24).Rats in group P were treated by probucol (110 mg·kg-1·d-1); rats in group D and group C were given equal volume water instead.Scr,BUN,triglyceride (TG),total cholesterol (TC) and 24-hour urinary proteinin were measured at the 4th,8th and 12th week.PAS staining and HE staining were used to evaluate the pathological changes of the kidney.The immunohistochemistry and Western blotting were used to detect the expression of 4-HNE in renal tissue.Results Levels of Scr,BUN,TG,TC and 24-hour urinary protein in group D were higher than those in group C at the 4th,8th and 12th week(all P ＜ 0.05); Levels of Scr,BUN,TG,TC and 24-huor urinary protein in group P were lower than those in group D at 4th,8th and 12th week (all P ＜ 0.05).The pathological changes of the kidney in group D were more serious than that in group P.The expression of 4-HNE in group D were higher than group C at the 4th,8th and 12th week (all P ＜ 0.05);The expression of 4-HNE in the kidneys of group P decreased significantly compared to that of group D at the same time (P ＜ 0.05).Conclusions As an indicator of lipid peroxidation,the expression of 4-HNE significantly increases in the kidney of diabetic rat.Probucol may protect the diabetic kidney through decreasing the expression of 4-HNE and the level of lipid peroxidation.

Objective To investigate the expression of glucose-regulated protein 78(GRP78)and cysteine aspartic acid protease 12(Caspase-12) and evaluate the endoplasmic reticulum stress (ERS) in rats with contrast-induced nephropathy (CIN),and observe the protective effects of hydroxytyrosol on CIN rats.Methods Eighty-four Wistar rats,(220±20) g,were randomly divided into control group,CIN group,hydroxytyrosol treated group (group C+H).At 12th,24th,48th,72th day after the rats model were established,BUN and Scr were detected.ELISA were used to detect the expression of methane dicarboxylic aldehyde (MDA),superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px).HE staining were used to evaluate the pathological change of kidney.TUNEL were used to detect the apoptosis of tubular ceils.Real-time PCR were used to detect the expression of GRP78 mRNA and Caspase-12 mRNA in tubular cells.Immunohistochemistry and Western blotting were used to detect the expression of GRP78 and Caspase-12 protein in tubular cells.Results BUN,Scr,the mRNA and protein expression of GRP78,Caspase-12 in hydroxytyrosol treated group were higher than that in control group(P ＜ 0.05),but were significantly lower than that in CIN group (P ＜ 0.05).Pathological changes and the apoptosis of tubular cells in CIN group were more serious than that in hydroxytyrosol treated group (P ＜ 0.05).Conclusions Endoplasmic reticulum stress may be associated with contrast-induced nephropathy.Hydroxytyrosol can protect kidney from contrast medium via reducing the endoplasmic reticulum stress.

OBJECTIVETo prepare tumor antigen peptides from HL-60 cells and to induce specific immune response.

METHODSHL-60 antigen peptides were obtained using techniques including freezing and thawing, heat precipitation and acid precipitation. The stimulating effect of the in vitro Hsp70 binding HL-60 peptides on PBMC and the proliferation of stimulated PBMC were observed by T cell activation test. The cytotoxicity of proliferated PBMC is detected by incubating HL-60 cells or K562 cells with PBMC respectively.

RESULTSThe obtained tumor antigen peptides were a peptides mixture. The mixed peptides could activate PBMC and cause PBMC proliferation in vitro after presented by Hsp70. The proliferated PBMC showed specific cytotoxicity to HL-60 cells but not to K562 cells.

CONCLUSIONThe method for preparing of human leukemia tumor antigen peptides used in this paper is simple and easy; the obtained antigen peptides can induce specific immune response in vitro.

Cell Division ; HL-60 Cells ; HSP70 Heat-Shock Proteins ; immunology ; Humans ; K562 Cells ; Killer Cells, Natural ; immunology ; Leukocytes, Mononuclear ; cytology ; immunology ; Neoplasm Proteins ; immunology ; Peptides ; immunology

Objective To discuss whether mild hyperuricemia can lead to kidney damage and the protection of decreased uric acid,through observing that hyperuricemia did damage to glomerulus endothelial function and cell proliferation of vascular smooth muscle in rats. Methods Fifty-four male SD rats were divided into four groups,the control group,model group (Oxonate),allopurinol group and Oxonate+allopurinol group.Rats were administered on a low sodium diet and their systolic blood pressure (SBP) were measured each 10 days.ELISA was used to detect rat plasma markers of endothelial function damage [nitric oxide (NO),type-1 plasminogen activator inhibitor (PAI-1),endothelin 1 (ET-1)] and cell proliferation of vascular smooth muscle[plateletderived growth factor (PDGF),cycloxygenase 2 (COX2),monocyte chemotactic protein-1 (MCP-1)],and the markers of inflammatory reaction[interleukin-18 (IL-18),tumor necrosis factor α(TNF-α)].PDGF and nitric oxide synthase (NOS) levels of rats were detected by immunohistochemical method.Renal tissue pathology of rats was observed. Results Compared to the control group,the plasmic concentration of COX2,ET-1,IL-18,PAI-1,PDGF,TNF-o,MCP-1 increased,and NO decreased significantly in rats of model group (all P＜0.05),expression of NOS significantly reduced and PDGF increased (all P＜0.05).Under light microscope,vascular wall thickening,intimal proliferation and lumen slight stricture without uric acid crystals in renal tissue were found in model group,which were obviously improved by using allopurinol. Conclusion Mild hyperuricemia can do damage to endothelial function of glomerulus and lead to vascular cell proliferation,which can be improved through decreasing uric acid.