Objective To assess the effect of suppression of insulin-like growth factor-1 receptor (IGF1R)in HO8910PM cell line by small interference RNA(siRNA).Methods Transfection of siRNA using liporectamine 2000 was conducted to silence IGF1R gene expression,the expression levels of IGF1R mRNA and protein were evaluated,and the effects on the cell cycles at 48 hours of transfection were assessed by real-time PCR,western blot and flow cytometry(FCM)assay respectively.The cell growth was detected by cell counting kit-8(CCK-8)at 24,48,72,96 hours of transfection.After 24 hours of transfection,the cells were cuhured with difierent concentrations of cisplatin(DDP)for 24 hours,the cell growth inhibition rate Was evaluated by CCK-8.Following incubation with 10μg/ml DDP for 24 hours after 24 hours of transfection,the apoptosis cells and the protein expression level of apoptosis-related gene,B cell leukemia/lymphoma 2(Bcl-2),were identified by FCM and western blot respectively.Resuits (1)Expression levels of IGF1R mRNA and protein were markedly decreased respectively at 48 hours of transfection IGF1R siRNA.(2)Suppression of IGF1R accompanied the reduction of cell growth at 48,72,96 hours of transfection with IGF1R siRNA,absorbance were 1.71±0.13,2.32±0.23,2.79±0.28 respectively (P＜0.01).(3)IGF1R siRNA induces arrest of G2 phase,the G2 phase rate of cells were 24.37%(P＜0.05).(4)Following treatment with 2.5,5,10,20μg/ml DDP for 24 hours after 24 hours of transfection,the cell growth inhibition rates were(25.94±0.08)%,(40.25±0.05)%,(59.48±0.03)%and(74.18±0.08)%respectively(P＜0.01).(5)Treatment with 10μg/ml DDP for 24 hours after 24 hours of transfection,induces 17.95%of cells apoptosis(P＜0.05),and decreases Bcl-2 protein level.Conclusion RNA interference of IGF1R gene induces the IGF1R silence in HO8910PM cell line significantly,inhibits cell growth in vitro, arrests the G2 phase, and enhances the chemosensitization to DDP.

Purpose:To study the effect of FasL gene on human lung adenocarcinoma cell lines and possibility of ex- ogenous FasL gene for gene therapy of lung cancer.Methods:An adenoviral expression vector with full length cDNA of FasL gene insert was constructed(Ad-FasL) and transfected into A549 cells.The effect of exogenous FasL gene on the growth of A549 cells was examined in vitro and in vivo.Results:Expression of FasL gene in A549 cells was confirmed by FCM and RT-PCR.The in vitro growth of the Ad-FasL transfected A549 cells was significantly inhibited (inhibition rate: 84%) as compared to mock (Ad-LacZ) transfected A549 cells.Colony-forming activity in vitro of the Ad-FasL transfected A549 cells was completely inhibited.The Ad-FasL transfected cells became apoptotic which was confirmed by the appear- ance of pre-G_1 on flow cytometry (FCM).The growth of A549 xenografts in nude mice was retarded by intra-tumol injection of Ad-FasL.Conclusions:FasL gene participates in the induction of cell apoptosis.Its use in gene therapy of cancer is promising.

Objective To evaluate the effectiveness of intravenous immunoglobulin (IVIG) in critical hand-foot-mouth disease (HFMD).Methods The data from PubMed, MEDLINE, EMBASE, EBSChost, Cochrane Library, Cochrane Central Register of Controlled Trials, Ovid, China Biology Medicine disc, Wanfang Data, China National Knowledge Infrastructure, Chinese Citation Database, and other references and grey literatures were retrieved, screening out all those related to clinical trials on treating critical HFMD by IVIG.Standard methods of the Cochrane Collaboration were employed to evaluate the methodological quality of the trials.Meta analysis was performed with Rev man 5.3 software.Results Eleven trials including 967 cases were investigated.The meta analysis showed that IVIG had significantly clinical efficacy (OR =6.84,95％ CI:3.74-12.52 ,P ＜ 0.05).IVIG could significantly decrease duration of fever (MD =-1.94,95％ CI:-3.07--0.81 ,P ＜0.05) ,hospitalization time (MD =-4.56,95％ CI:-8.95--0.17,P ＜0.05).There was no significant difference in duration of fever (MD =-0.28,95 ％ CI:-0.59-0.03, P ＞ 0.05), duration of herpes (MD =0.18,95％ CI:-0.22-0.59, P ＞ 0.05), hospitalization time (MD =-0.12,95％ CI:-0.47-0.23, P ＞ 0.05) when the dosage of injection was adjusted.Conclusions IVIG is recommended for treating critical HFMD because it is effective in decreasing the duration of fever and hospitalization.Well designed studies with more sample in multi-center are required in further study to explore the efficacy and safety of IVIG on critical HFMD.

Adolescent ; Adult ; Child ; Elliptocytosis, Hereditary ; diagnosis ; genetics ; Female ; Humans ; Male

To prepare the RNAse-resistant virus particles containing the partial gene fragments of avian influenza virus H5N1 for use as RNA standard and control in RNA virus detection, the genes coding the coat protein and maturase of E.coli bacteriophage MS2 were amplified by PCR and then cloned into prokaryotic expression vector pET32a to construct the intermediate vector pET32a-MS2. In addition, the gene sequences coding hemagglutinin (HA), neuraminidase(NA) and M protein of the H5N1 virus were also cloned separately to the down-stream of plasmid pET32a-MS2, thus constructing the prokaryotic expression vectors pET32a-NS2-HA, pET32a-MS2-NA and pET32a-MS2-M. These recombinant plasmids were then transformed separately to E.coli BL21(DE3) with induction by IPTG. to express the virus-like particles. The virus-like particles observed under electron microscopy were identified by RT-PCR ,while their stability was confirmed by real-time RT-PCR. In this way, the virus-like particles were successively constructed and identified through PCR amplification, enzymolysis identification and sequencing analysis. These virus-like particles observed under electron microscopy appeared to be circular in shape with a diameter of about 50 nm. Their stability was proved to be rather good. From these observations, it is apparent that these virus-like particles can be used as RNA standard and quality control in the detection of avian influenza virus H5N1.

Objective To compare the glucose levels and associated factors among the normal glucose tolerance subjects with different age.Methods Totally a community-based population of 2098 residences aged above 30 years Were tested with OGTT,and classified into normal glucose tolerance group(NGT),impaired glucose tolerance group(IGT),impaired fasting glucose group(IFG),both IGT and IFG group(ICT/IFC),anddiabetes group(DM) according to fasting and 2 hours glucose level(2 hPG).The subjects in NGT group were further divided into 5 groups according to different ages.The levels of blood glucose and HBCI in different groups and subgroups were measured and analyzed statistically. Results For patients in NGT,the FPG([5.17.±0.48]mmol/L vs.[5.09±0.44]mmol/L,P<0.05)and HbA1c([6.01±0.62]%vs.[5.95±0.66]%.P<0.05)in group aged 60-69 Were higher than that in group aged 50-59.The FPG in group aged 60-69 was also higher than those in group aged 40-49([5.17±0.48]mmol/L vs.[5.00±0.47]mmol/L,P<0.01),and the FPG in group aged 50-59 Was also higher than those in group aged 40-49([5.09±0.44]mmol/L vs..[5.00±0.47]mmol/L,P<0.01).There was no correlation between age and FINS,while a tendency of decreasing HBCI could be observed along with increasing of age(F=33.75,P<0.05).Conclusion In NGT subjects,the FPG and HbA1 C inereased along with age.

Objectives To investigate the expression and diagnostic value of 14-3-3 protein in brains of patients with sporadic Creutzfeldt-Jakob disease(sCJD).Methods 14-3-3 protein was immunohistochemically analyzed in tissue from the frontal lobe of 5 patients with sCJD and 4 non-CJD eases Using 14-3-3 ?and ?antibodies with reference to the results of KB,GFAP and PrP detection.Results The expressions of 14-3-3 protein in five brains of sCJD were more obviously,mostly in gray matters and astrocytes in three cases.The concentration was related to PrP deposition type,but not related to prion protein genotype.Except few expression of 14-3-3 protein in neurous of two cases of acute contusion,there were no expression in the other two cases in control group.Conclusions The expression of 14-3-3 protein in brain is useful to pathological diagnosis of CJD.

Objective To observe effects of Naotaifang on MMP-9, NF-κB and TIMP-1 after focal cerebral ischemia in rats. Methods The rats were randomly divided into sham operation group, model group, and Naotaifang low- (3 g/kg), medium- (9 g/kg), high- dose (27 g/kg) group. After 3 days of corresponding therapy by intragastric administration once a day, the regional cerebral ischemia model was made by middle cerebral artery occlusion (MCAO) with suture method. Following 3 days, the rats were treated with previous method. On the third day, hippocampal C2 region of ischemic tissue was detected by HE dyeing. And the contents of MMP-9, NF-κB and TIMP1 proteins in hippocampal C2 region were measured by immunohistochemical method. Results The number of normal brain cells in high dose group of Naotaifang was more than that of the model group, and only a few cells appeared nucleus pycnosis. The MMP-9 expression of all dose groups of Naotaifang were significantly decreased than model group (P<0.05). The NF-κB expression of high and medium dose groups of Naotaifang were significantly decreased (P<0.05). The TIMP1 expression of all dose groups of Naotaifang were significantly increased compared with sham operation group (P<0.05). Conclusion The mechanism of Naotaifang protecting blood brain barrier against injury of cerebral ischemia may be involved in ameliorating MMP, NF-κB and increasing TIMP1 expression.

OBJECTIVETo examine the effect of ginsenoside Rb1 (GRb1) on the immune maturation of monocyte-derived dendritic cells (DCs) induced by oxidized low-density lipoprotein (OX-LDL).

METHODSHuman monocytes purified by CD14+ immuno-magnetic beads were differentiated and induced into immature DCs, which were randomly divided into 6 groups, Group A treated with PBS, Group B treated with OX-LDL, Group C and D treated respectively with GRb1 and ciglitazone, Group E and F were pretreated with the two testing drugs respectively followed by OX-LDL. The immuno-phenotypic expression (CD40, CD1a, and HLA-DR) and endocytosis function of DCs were examined using flow cytometry, the concentration of interleukin-12 (IL-12) and tumor necrosis factor alpha (TNF-alpha) in the culture supernatants were measured with ELISA.

RESULTSCompared with Group B, Group E showed significantly lowered immuno-phenotypic expression of DCs in terms of CD40 (67.4 +/- 1.62 vs. 145.69 +/- 14.86), CD1a (79.64 +/- 3.04 vs. 159.89 +/- 6.09), and HLA-DR (46.43 +/- 2.85 vs. 99.33 +/- 17.11), as well as higher endocytosis level (88.13% +/- 1.06% vs. 25.90% +/- 5.77%, all P < 0.01). Meantime, the serum levels of IL-12 (88.65 +/- 5.59 ng/L vs. 716.69 +/- 36.35 ng/L) and TNF-alpha (133.27 +/- 11.98 ng/L vs. 968.10 +/- 36.42 ng/L) obviously decreased (P < 0.01). The surface molecular expression of DCs and the secretion of inflammatory factors in Group F also obviously decreased, showing insignificant difference from Group E (P > 0.05).

CONCLUSIONGRb1 could obviously inhibit the OX-LDL-induced maturation of DCs, showing similar effects to ciglitazone.

Cell Differentiation ; drug effects ; Cells, Cultured ; Dendritic Cells ; cytology ; drug effects ; immunology ; Flow Cytometry ; Ginsenosides ; pharmacology ; Humans ; Lipoproteins, LDL ; Monocytes ; cytology ; drug effects ; immunology

Objective To investigate the effects of transarterial chemoembolization (TACE)treatment interval on the prognosis of patients with advanced hepatocellular caisinoma(HCC).Methods We retrospectively collected clinical data of 123 advanced HCC patients treated with repeated TACE.The patients were divided into two groups (group A with fixed repeated treatment interval and group B with that according to the clinical needs).Cox regression,survival curve and log-rank test were used to assess the effects of the treat-ment intervals on prognosis.Results The treatment intervals of the group A and group B were (1.1±0.3)months and (3.0±1.5) months,respectively (P <0.001).Multivariate Cox analysis showed the efficacy (P =0.024)and repetition periods (P <0.001 ) were independent prognostic factors.Conclusion TACE interval is independent risk factor for the prognosis of patients with ad-vanced HCC.Repeated TACE treatment according to clinical needs may be more favorable for prognosis of the patients.