Objective:To investigate the availability for rhIL-18 in the in vitro culture system to stimulate PBMHCs,with inducing cytotoxicities against tumor cells and to analyze the influencing factors for the effects.Methods:The NK cells,T cells and dendritic cells were separated from fresh PBMNCs by using the Stem Sep TM immunomagnetic beads.Cell phenotypes of the purified cell populations were identified with FCM technique.The cell co-culture system was established as follows.The PBMNCs or the cell preparations deleting the definite cell subset with immunoscreening were co-cultivated with mitotic-inactivated tumor cells in the presence of rhIL-18.Cytotoxicities of the various effector cell preparations to a series of tumor cell lines were examined by the isotope releasing assay.Results:In the in vitro cell co-culture system,rhIL-18 rapidly induces activation of the cytolytic responses against various tumor cell lines mediated by PBMNCs.The rapid induction within 24 h of culture was dependent on the dosage of rhIL-18,with the optimal dose of 100 ng/ml of the cytokine.The activated cytotoxicities were abrogated by deleting NK cells prior to the cell co-culture but did not vary when either T cells or DCs were removed.The cytotoxic responses were shown as a pattern of broad-spectrum to the target cells used and were not blocked by anti-MHC moAbs.Conclusion:NK cells were responsible for the rapid induction of cytotoxicities against tumor cells by rhIL-18.

Objective:To investigate the expression of microRNA-199a/b-3p (miR-199a/b-3p) in hepatocellular carcinoma ( HCC) tissues,and to explore the relationship with clinical outcomes. Methods: Real time quantitative PCR technique was used to measure the expression of miRNA-199a/b-3p in HCC tissues. The correlation between miR-199a/b-3p expression and the clinic pathological features of patients were analyzed. Results: Comparing with adjacent control, miRNA-199a/b-3p presented lower expressions in HCC tissues (P<0. 05);lower miR-199a/b-3p was found correlated with metastasis and poor survival. Conclusion:MiR-199a/b-3p take a crucial role in HCC metastasis and recurrence.

Objective To investigate the effect of tannic acid on glomerular mesangial cells (GMC),and to clarify the mechanism of tannic acid in improving the pathological changes of diabetic nephropathy (DN)from the aspect of oxidative stress and micro-inflammation. Methods The glomerular mesangial cells were treated with glucose (30 mmol·L-1 )or advanced glycosylation end-products (AGEs)bovine serum albumin(BSA)(250 mg·L-1 )and then different concentrations of tannic acid (10,20,40 and 80μmol·L-1 )were added into the GMC.The cells cultured by normal glucose or treated with BSA were used as control groups and then the level of malonic dialdehyde (MDA), glutathione peroxidase (GSH-Px ), superoxide Dismutase (SOD ), CAT (Catalase ) activities and 8-hydroxy-2′-deoxyguanosine(8-OHdG)levels in the culture supernatant 48 h after culture were determined by colorimetry and ELISA method. The expressions of intercellular cell adhesion molecule-1 (ICAM-1 ) protein, monocyte chemotactic protein 1 (MCP-1 ) and ICAM-1 mRNA in GMC were detected by immunohistochemical staining and RT-PCR method.Results Compared with high glucose and AGEs groups,the MDA levels in tannic acid groups were reduced significantly(P<0.05);the activities of GSH-Px,SOD and CAT were increased significantly(P<0.05 or P<0.01);the 8-OHdG levels in annic acid groups were significantly reduced (P<0.05). Compared with high glucose and AGEs groups,the expressions levels of ICAM-1 protein in 40 and 80μmol· L-1 tannic acid groups were decreased (P<0.05 ). The mRNA expressions levels of MCP-1 and ICAM-1 were significantly lower than those in high glucose group (P<0.01 ).Conclusion Tannic acid could protect GMC against the damage of oxidative and inflammatory mediators,thereby delaying and improving the glomerular lesions of DN.

Objective To construct prokaryotic expression plasmid of human recombinant soluble TNF-related apoptosis inducing ligand(rsTRAIL),then to investigate the effects of rsTRAIL on apoptosis in non-small cell lung carcinoma(NSCLC).Methods The encoding sequence for rsTRAIL was amplified with RT-PCR and cloned into PQE30 vector to establish the prokaryotic expression system.The competent cells of host strain of M15 were transformed by the recombinant plasmid.The expression of the target protein was induced with IPTG and purified by Ni2+-NTA agarose column.rsTRAIL was added in the media of A549 and H460wt cells,then the viability was examined by MTT assay.Apoptosis of H460wt cells was observed under fluorescence microscopy.The apoptotic rate of tumor cells was examined by FACS.Results The cloned fragment of rsTRAIL was 100% consistent with that reported in GenBank.The expressed protein with molecular weight of 21 000 in SDS-PAGE as expected was obtained and recognized by a commercial McAb.The apoptotic rate of H460wt cells after treated with rsTRAIL for 24 h was 43.2%.Cislatin enhanced the effect of rsTRAIL on A549 cells.Conclusion The rsTRAIL is obtained after Ni affinity chromatograph.rsTRAIL has a strong cytotoxic activity against NSCLC and cisplatin may enhance the antitumor effect of rsTRAIL.

Objective To construct a recombinant expression vector of human interleukin-24(hIL-24) gene and express it in E.coli M15,and to evaluate the bioactivity of IL-24 fusion protein.Methods The human IL-24 cDNA fragment was amplified from plasmid by polymerase chain reaction(PCR),and sequenced.PQE/hIL-24 was constructed by gene rearrangement,then it was transformed into E.coli M15.The expression of the target protein was induced with IPTG and purified by Ni2+-NTA agarose column.The expressed recombinant hIL-24(rhIL-24) was identified by SDS-PAGE and Western blotting.Normal peripheral blood mononuclear cells(PBMCs) were cultivated with the expression protein for 48 and 72 h,the levels of IL-6,IFN-? and TNF-? of PBMCs stimulated with rhIL-24 were detected by ELISA.Results The recombinant prokaryotic expression vector PQE/IL-24 was constructed successfully and expressed stably in E.coli M15.At about 18 400 of molecular weight,there was an induced protein band.The levels of IFN-?,IL-6 and TNF-? in the group of cultivated with the expression protein were obviously higher than those in the groups without the expresson protein(P

With the progress of visualization technology, more and more visual tools were applied to anesthesia. It has accurate positioning, mild trauma, a high success rate and less complications for endotracheal intubation, which increase the safety and efficiency of airway management. Disposcope endoscope is an endotracheal intubation tool with a visual stylet, which has many advantages, such as visual, lens barrel can be bent and wireless transmission. This paper summarizes the current progress in the application of Disposcope endoscopy in tracheal intubation.

Objective To investigate the effects of topiramate on neurons apoptosis and the hydrous capacity in brain tissue in rats with perinatal acute ischemic brain damage.Methods The perinatal acute ischemic brain damage model was established by ligation of both uterine arteries of full-term pregnant Wistar rats(n=315).They were divided randomly into topiramate one and five times groups and hypoxic-ischemia(HI) group(n=105),and normal collate group consisted of 105 normal Wistar rats.The effects of topiramate were observed by the changes of hippocampal apoptosis cells labeled in situ end TUNEL methods at different time spots(postnatal 3 h,6 h,24 h,(3 d,7 d),14 d,21 d,28 d) and the hydrous capacity in brain tissue within 7 d after birth.Results The hydrous capacity in brain tissue in administering drug one time group was remarkably less than that in HI group at 12 and(72 h after) birth,and it in administering drug five time group was remarkably less than those in HI group and administering drug one time group at 48 and 72 h after birth(P

Objective To study the inhibitory effects of dehydroepiandrosterone (DHEA) and its metabolites-dehydroepiandrosterone sulfate (DHEAs) on the proliferation of HepG2 and HT-29 and their mechanism.Methods HepG2 and HT-29 were incubated by DHEA or DHEAs with different concentrations (1,10,50,100 and 200 ?mol?L-1) for 8,24,48,72 h and routine culture was used as control.The inhibitory rate was detected by using MTT chromometry and BrdU assay respectively.The activities of 3-hydroxy-3-methylglutaryl coenzyme A reductase(HMGR),glucose -6-phosphate dehydrogenase (G6PD) and lactate dehydrogenase (LDH) were examined simultaneously.Results ①MTT chromometry:DHEA with different concentrations obviously inhibited the growth of HepG2 and HT-29 cells compared with control group(P0.05).②BrdU assay:the growth of cells were significantly inhibited by DHEA with concentrations of 50,100 and 200 ?mol?L-1,especially to HepG2 cells(P0.05).Conclusion DHEA has strong anti-proliferative effects on both HepG2 and HT-29 cell lines and inhibitory effects on the activities of G6PD or HMGR,however,DHEAs has no obvious effect.

Objective To investigate the relationship between the expressions of P16 and proliferating cell nuclear antigen(PCNA) in osteosarcoma and clinicopathological features,and explore the effects of them in occurrence and development of osteosarcoma.Methods The expressions of P16 and PCNA were detected by immunohistochemistry(SP) in 71 osteosarcima tissues and 10 normal bone tissues.Results ①The positive rate of P16 expression in osteosarcoma tissues was lower than that in normal bone tissues(P0.05).The expression of PCNA had positive relationship with osteosarcoma's prognosis(P0.05).③There was negative correlation between P16 and PCNA expressions(rs=-0.58,P

Objective To investigate the suppression to 2-glycoprotein-1-specific B cell response by human recombinant 2-glycoprotein-1 domain Ⅰ dimer(rh?2-GP1-DⅠ2) in rh?2-GP1 immunized mice.Methods The effects of treatment using rh?2-GP1-DⅠ2 on titer and ratio of anti-?2-GP1 were analyzed by solid phase ELISA.The number of splenic ?2-GP1-specific antibody-forming cells(AFC) was counted by ELISPOT.Results The levels of anti-?2-GP1 from rh?2-GP1 immunized mice treated with rh?2-GP1-DⅠ2 were significantly decreased compared with those in negative controls(P