Aged ; Amyloidosis ; therapy ; Humans ; Male ; Medicine, Chinese Traditional ; Moxibustion ; methods ; Skin Diseases ; therapy

Objective To evaluate the immune response triggered by an in-house constructed hu-man metapneumovirus multi-epitope antigen ( MEA) in a mouse model .Methods Female SPF BALB/c mice at age 4-6 weeks were used in the study and divided into 7 groups.Mice in the five groups including MEA+oligodeoxynucleotides containing CpG motifs ( CpG ODN) intraperitoneal injection ( i.p.) treatment group, MEA+Alum i.p.treatment group, MEA+Alum+CpG ODN i.p.treatment group, MEA+CpG ODN intranasal (i.n.) treatment group and MEA+Alum+CpG ODN i.n.treatment group were immunized three times on days 0, 14 and 21, and those in the other experimental group were immunized intramuscularly with MEA+Quickantibody5W on days 0 and 21.A control group without treatment was set up accordingly .All mice were sacrificed two weeks after the last immunization .Antibodies including IgG , IgG1, IgG2a and IgA in serum samples were detected by ELISA .MTS assay was performed to analyze the proliferation of lympho-cytes.The cytotoxicity of cytotoxic T lymphocytes (CTL) was measured by LDH assay.Flow cytometry was used to detect T lymphocyte subsets .The cytokines secreted by T helper cells ( Th1 and Th2) were analyzed with Bio-Rad Liquid Chips.Results High titers of IgG, IgG1 and IgG2a antibodies were produced in MEA treated mice except for those in intranasal treatment groups .Serum samples from three groups including the MEA+Alum i.p., MEA+Alum+CpG ODN i.p.and MEA+Quickantibody5W i.m.treatment groups were positive for IgA antibody .The highest titer of IgA antibody was detected in mice from the MEA+Alum+CpG ODN i.p.treatment group, which was 2.15×103.Compared with the control group, significantly enhanced proliferation of lymphocytes was observed in the MEA+Alum i.p., MEA+Alum+CpG ODN i.p.and MEA+Quickantibody5W i.m.treatment groups (P<0.05).Enhanced cytotoxic activities of CTL were observed in mice with ip.and i.m.treatments as compared with those in control group (P<0.05).The levels of CD4+/CD8+T cells were slightly increased in mice from the MEA+CpG ODN i.p., MEA+Alum+CpG ODN i.p. and MEA+Quickantibody5W i.m.treatment groups as compared with those in control group (P<0.05).In-creased secretion of IL-2, IFN-γand Th2-type cytokines including IL-4, IL-5 and IL-10 were detected in mice from the MEA+CpG ODN i.p.treatment group.The MEA+Alum i.p.treated mice showed a slightly increased secretion of IFN-γand significantly increased secretions of IL-4, IL-5 and IL-10.Significantly in-creased secretions of IFN-γ, IL-4, IL-5 and IL-10 were detected in mice from the MEA+Alum+CpG ODN i.p.treatment group.Significantly increased secretions of IFN-γ, IL-5, IL-10 and granulocyte-macrophage colony-stimulating factor (GM-CSF) were detected in mice from the MEA+Quickantibody5W i.m.treatment group.Conclusion MEA together with different adjuvants could stimulate high titers of specific antibodies , increase the proliferation of lymphocytes and enhance the cytotoxic activities of CTL .CpG ODN could bal-ance the Th1/Th2-mediated immune responses , and the balance could be enhanced when using CpG ODN in combination with Alum .A similar effect could be achieved by using the commercial adjuvant Quickanti -body5w.This study has paved the way for further investigation on the development of hMPV epitope vaccines and diagnostic reagents for hMPV as well as the epidemiological study of hMPV .

0.05).There was a significant positive correlation between serum VEGF,NP and the degree of proteinuria in group A patients,respectively(r=0.47,0.43 Pa

Objective To detect the levels of serum neopterin(NP) and D-dimer,and explore whether NP and D-dimer levels respond to coronary artery dilatation(CAD) of Kawasaki disease(KD),and evaluate the risk factors of CAD.Methods This study were conducted on 45 children with KD.Twenty-one children were CAD and 24 cases were not CAD.Active phase serum NP,D-dimer levels were measured by sensitive enzyme-linked immunosorbent assay(ELISA).Multiple regression analysis including platelet(PLT),monocyte(M),creatine kinase-MB(CK-MB),C-reactive protein(CRP),erythrocyte sedimentation rate(ESR),cardiac troponin I(cTnI) were studied with CAD.Results Compared with recovery phase,active phase the levels of PLT,M,CK-MB,CRP,ESR,cTnI were significantly higher(Pa

Background:Recently,cancer stem cells( CSCs)has become a hot topic in cancer research. For this research topic,the primary problem is the isolation and identification of CSCs,and suspension culture is considered to be an important method for CSCs isolation. Aims:To study the isolation and identification of gastric cancer stem cells( GCSCs) and evaluate its stemness characteristics. Methods:GCSCs-like cells were enriched through spheroid formation by suspending the human gastric cancer cell lines MKN45,MGC803 and SGC7901 in serum-free medium containing epidermal growth factor( EGF)and basic fibroblast growth factor( bFGF). The third generation tumor spheres obtained were tested for the ability of colony forming;its stemness markers were identified by immunofluorescent staining,and migration and invasion capabilities were assessed by wound scratch test and Transwell assay,respectively. Growth inhibition of tumor spheres in response to cisplatin was assessed by CCK-8 assay. Results:High expressions of stem cell markers CD44 and CD54 were observed in the tumor spheres,but gastric parietal cell marker H+ /K+-ATPase was lowly expressed. Compared with parental gastric cancer cells,tumor spheres had stronger colony forming ability(P<0. 05);Furthermore,healing of wound scratch was accelerated(P<0. 05),and number of invasive cells in Transwell chamber was increased in tumor spheres(P<0. 05). The 50% inhibition concentration of cisplatin to tumor spheres increased significantly when compared with parental gastric cancer cells. Conclusions:GCSCs enriched tumor spheres can be obtained by suspension culture with serum-free medium. The tumor spheres possess enhanced capabilities of self-renewal,proliferation,migration,invasion and potential of multilineage differentiation;meanwhile,it is more resistant to chemotherapy.

0.05).The change of heart rate in 2 groups was greater than that of distilled water respectively(P

Objective To investigate expression of transforming growth factor-?1(TGF-?1)and connective tissue growth factor(CTGF) protein in renal tissues,and detect the levels of urinary TGF-?1 and CTGF in rats with diabetic nephropathy(DN).To observe the influence of losartan on expression of the two protein in renal tissue and excretion in urine.Methods Wistar rats were treated by intravenous injection of streptozotocin after right nephrectomy to induce DN rat model.The DN rats were randomly divided into two groups:DN experimental group and losartan treated group.The expression of TGF-?1 and CTGF in renal tissue were determined with immunohistochemical staining,urinary TGF-?1 and CTGF were assayed by enzyme-linked immunosorbent assay(ELISA) at 6,12 weeks respectively.Results Compared with losartan treated group,urinary protein excretion and the protein expression of TGF-?1 and CTGF significantly increased(P

Objective To detect the level of serum cystatin C(CysC),macrophage migration inhibitory factor(MIF),urinary podocytes(UPC) and explore the correlation between the levels of the three and therapeutic effect in children with primary nephrotic syndrome(PNS).Methods The serum CysC,MIF levels were detected in 38 children with PNS,which were treated before and after remission by double-antibody sandwich sensitive enzyme-linked immunosorbent assay(ELISA).Podocyte-specific marker protein-Podocalyxin(PCX) was detected by immunofluorescene to definite UPC before and after remission and compare with those of healthy control group(n=15).Results Compared with control group,the levels of serum MIF were significantly increased in 22 patients responded sensitively to prednisone prior and post treatment(Pa0.05),however,CysC,MIF and UPC significantly increased after treatment in 16 steroid-resistant children(Pa0.05), MIF significantly decreased(Pa0.05)in steroid-resistant children after treatment.In 16 children with steroid-resistant, complete remission of PNS as cyclophosphamide pulse treatment being added in 10 patients,which CysC, MIF,UPC were significantly higher than those in control group(Pa

Objective To explore the expression of vascular endothelial growth factor(VEGF) and macrophage migration inhibitory factor(MIF) in kidney of diabetic rats,the influence of protein kinase C ? inhibitor LY333531 on the expression of VEGF and MIF.Methods Thirty wistar rats were divided at random into three groups:normal control group(NC group);diabetic model group(DM group),the rats models of diabetic mellitus were induced by streptozotocin(SIZ);LY333531 treated group(LT group),which were given LY333531 [10 mg/(kg?d)] by gastric perfusion after DM model inducing success.At the 10th week,the levels of blood glucose(BG),24 hours urine albumin excretion rate(AER),urine VEGF,and MIF were detected using enzyme-linked immunosorbent assay(ELISA).Glomerular cellularity and extracellular matrix(ECM) were observed by light microscope.The expression levels of VEGF and MIF in the kidney tissues in three groups of rats were detected using immunohistochemistry.Results BG,AER,VEGF,MIF in DM group were significantly higher than those in control group(Pa0.05).There was strongly stained of VEGF,MIF in glomerular mesangium in DM group,weakly positive in LT group by immunohistochemistry.The expression of VEGF and MIF in DM gorup significantly increased than those in NC group(P0.05).Conclusions The expression levels of VEGF and MIF in kidney tisues of diabetic rats significantly increase and correlation with the degree of renal tissue injury.LY333531 can contribute to cut down BG,decrease AER,VEGF,MIF excrection in urine,inhibit glomerular cellularity and ECM proliferation,inhibit expressions of VEGF and MIF to protect kidney.

Objective To explore the effect of 1,25-(OH)2D3 on Nephrin protein expression in renal of diabetic rats.Methods Diabetic rats were induced by streptozotocin(STZ).Groups were divided as follows:diabetic group(DM group),1,25-(OH)2D3 treated group[1,25-(OH)2D3 group] and control group.At the 12th week,haematuria and proteinuria were determined and the kidneys were removed by histopathology,immunostaining,and the expression of Nephrin mRNA and protein in renal cortex were detected by reversed transcription-polymerase chain reaction(RT-PCR).Results Compared with control and 1,25-(OH)2D3 groups,urine erythrocyte and protein excretion and glomerlar cellularity in DM group statistically increased(P0.05).Conclusion The effect of 1,25-(OH)2D3 is in relieving haematuria and proteinuria and up-regulate the expression of nephrin mRNA and protein to protein kidney.