Objective To investigate the immunophenotype and the cytotoxicity of cytokine-induced killer(CIK) against tumor cells in vitro.Methods Lymphocytes cells were isolated freshly from peripheral blood of healthy donors by Ficoll-Hypaque density centrifugation,and the cells obstained were induced by IFN?,IL-2 and CD3McAb.Phenotypes and cytotoxicity of CIK were analysed by FACS.Target cells were differentiated from effect cells by CFSE dying.The mortality of Target cells were determined by FACS.Results The maximum proliferation of CIK reached at the 22nd day.The phenotypes of CD3,CD11a,CD54,HLA-DR were expressed highly;CD25,CD28,CD69,FasL were expressed moderately on CIK.The expression of CD16 was not increased.CIK possessed the cytotoxicity against tumor cells of K562,HL-60,Hela,SMMC7721 and A375.Conclusion IFN?,CD3McAb,IL-2 can induce peripheral lymphocytes to produce CIK which own strongly proliferation and exert highly efficient cytotoxic effects on tumor cells.

Epithelial-mesenchymal transition (EMT) is the basis of biological process of embryonic development,and is also closely related with tumor cell in situ invasion and distant metastasis.Recent studies find that EMT and cancer stem cells (CSCs) have a close relationship.CSCs have a strong ability to selfrenewal,tumorigenicity and cell differentiation potential.Identifing the markers,in-depth study of CSCs resistance,may provide a new path for cancer treatment.

Objective To study the change of BDNF/NF-κB signal pathway induced by repeated febrile seizures.Methods Fifty-one 21 d male rats were randomly divided into three groups:normal control group(n=14),hyperthermal control group(n=19)and febrile seizure group(n=18).The febrile seizure model was devdoped by warrn water immersion[(45.0±0.3)℃].BDNF and NF-κB were measured by enzyme hnked immunosorbent assay(ELISA)and immunohistochemistry.Results The levels of BDNF in serum and hippocarnpus in FS group were higher than those in NC group(P＜0.01)and in HC group(P＜0.01).In FS group,the OD of the BDNF positive neuron was higher than that of NC group(P＜0.01)and HC group(P＜0.01);the OD of the NF-κb positive neuron in FS group was higher than that of NC group (P＜0.01)and HC group(P＜0.01),the OD of the NF-κB positive neuron in HC group higher more than in NC group(P＜0.01).There was positive correlation between BDNF expression and NF-κB activation(r=0.78.P＜0.01).Conclusion The expression of BDNF and NF-κB were obviously increased in FS group and they are positively correlated,suggesting that the BDNF/NF-κB signal pathway may be activated after febrile seizure.

ObjectiveTo investigate the relationship between the polymorphisms of the promoter of a disintegrin and metalloproteinase 10(ADAM10) gene and sporadic Alzheimer's disease (SAD).Methods The promoter of ADAM10 gene in 10 controls and 10 SAD patients was sequenced.Three variations were found,then these variations in 298 SAD patients (SAD group) and 315 healthy controls (control group)were genotyped by using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).ResultsThree polymorphisms were found in the promoter of ADAM10 gene: -279G/A (rs653765),- 630G/T( rs514049 ) and - 921GAGA/- ( rs33926666 ).For - 921GAGA/-,there were significant differences in genotype ( GAGA/GAGA:138 (46.3％ ),GAGA/-:155(52.0％),-/-:5(1.7％))and allele frequencies (GAGA:431 (73.6％),-:165 (27.7％) ) between SAD and control (genotype:x2 =34.130,P =0.000; allele:x2 =25.972,P =0.000). For - 279G/A,there were significant differences in genotype and allele frequencies between SAD and control in the subjects without ApoEε4 allele (genotype:x2 =8.734,P=0.013; allele:x2 =5.129,P=0.024). -279G and -921GAGA were relatively protective allele types for SAD,and they were not in linkage disequilibrium.ConclusionThe polymorphisms - 279G/A and - 921GAGA/- of ADAM10 are associated with SAD.Allele G or genotype G/G of -279G/A and the GAGA/GAGA genotype or the GAGA allele of -921GAGA/- might have a protective effect on SAD.

Abstract Objective: To prepare high-purification and high-specific activity of recombinant human interleukin-6 (rhIL-6). Methods: The rhIL-6 was obstained from inclusion body expressed by IPTG-induced pT7.7hIL-6 expressed vector using extracting,denature and refolding techniques. The rhTL-6 was further purified by anionic-exchange chromatography. Activity of rhIL-6 was measured by 3H-TdR method. Results: After a single-step purification,the product purity reach 95% and it's specific activity was 3.0 x 10~8 U/mg. Conclusion:This scheme of puri-fication was an easy way requiring rhTL-6.

Glottis ; pathology ; Humans ; Laryngeal Neoplasms ; surgery ; Minimally Invasive Surgical Procedures

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

Objective To get the protein expression of sFlt-1 by transfection and to investigate the effects of sFlt-1 gene transfection on the growth of K562 cells.Methods The recombinant plasmid pcDNA3-sFlt-1D4 was constructed.The recombinant plasmid pcDNA3-sFlt-1D4 was transfected into the bone marrow stromal cells by Lipofectamine 2000,which was identified by RT-PCR,ELISA and MTT.Results The transfection efficiency identified by flow cytometry was 9.27%.The protein expression of sFlt-1D4 was found in the culture supernatant 24 h,48 h and 4 weeks after transfetion by ELISA and the expression concentrations were(0.104?0.078),(0.158?0.022) and(0.171?0.069) ?g?L-1,respectively.The content of VEGF secreted by K562 culturing with transfectant cells culture supernatant was reduced compared with control.The inhibitoy rates on the proliferation of K562 cells via MTT assay were 9.41%?4.71%,23.63%?7.50%,and 33.13%?6.93%,respectively.Conclusion The bone marrow stromal cells transfected with recombinant plasmid pcDNA3-sFlt-1D4 could secrete sFlt-1D4 and inhibit the proliferation of K562 cells.

【Objective】To observe the hemorrheological changes in lacunar cerebral infarction(LCI) patients with arteriopathy of middle and small arteries.【Methods】The hemorrheological changes of cerebral arteries in 85 LCI patients without arteriopathy of large artery confirmed by cervical vessels color ultrasonography and magnetic resonance image(MRI)were observed by transcranial Doppler ultrasonography(TCD).The correlation of the hemorrheological changes with age and blood pressure(BP) of arteries was investigated.The above indexes were compared with those in 25 healthy volunteers confirmed by encephalography in the same age group.【Results】The mean blood flow rates of cerebral middle,anterior and posterior artery in LCI patients were(62.09?16.90)cm?s~(-1),(50.42?13.11)(cm?s~(-1),)and(32.33?7.55)cm?s~(-1) respectively,lower than(69.65?19.20)cm?s~(-1),(57.75?16.75)cm?s~(-1),and(38.75?8.81)cm?s~(1) respectively in the healthy volunteers(P0.05).【Conclusion】The arteriopathy of small artery in LCI patients can cause the mild decrease of blood flow rate and the increase of pulsating index.There exists a correlation of pulsating index with age and arterial blood pressure.

Objective To study the relationship between vascular obstruction,inflammation and the pseudolobuli formation in liver tissues with chronic hepatitis B.Methods The liver biopsies from 1256 patients with chronic hepatitis B was observated by HE,histochemical,immunohistochemical and the electricity mirror techniqe.Results (1)The intrahepatic vascular inflammation and obstruction was a histopathological characteristics of liver cirrhosis of chronic hepatitis B;(2)The lesions of vascular inflammation caused the vascular obstruction and induced the proliferations of venous and fiber;(3)The blood vessel of hyperplasia revolved round the liver cell into the irregular cell circular(tiny node).The tiny node settled the composition base in the interest of pseudolobuli formation;(4)The blood vessel of hyperplasia once more inflammation,blocking,fiber deposit constituted the fiber brings,pseudolobuli formation.Conclusion The vascular obstruction,inflammation has a closely relationship with the pseudolobuli formation in liver tissues with chronic hepatitis.