To study on the phylogenetic characterization of the VP1 genes of coxsackievirus A16 (CVA16) causing hand-food-mouth disease (HFMD) isolated from Anhui province in 2014. A total of 413 throat swab specimens from HFMD patients were collected during January to November, 2014 for the isolation and identification of enteroviruses using real-time RT-PCR assays. The VP1 regions of CVA16 isolates were amplified using RT-PCR and sequenced. And the phylogenetic tree was constructed among the VP1 regions of those isolates, the different genotypes and sub-genotypes of CVA16 strains. A total of 97 enteroviruses were isolated from 413 samples, the positive rate was 23.49% (97/413), including seventeen CVA16, seventy six HEV71 and four other enteroviruses. The results of the phylogenetic tree showed that 17.CVA16 strains isolated from Anhui in 2014 clustered within B1b evolution branch of B1 genotype. The nucleotide and amino acid sequence identities were 95.30%-100% and 98.70%-100% among the isolates, respectively, but within B1b branch of 17 strains formed several small transmission chains. The nucleotide acid of 17 CVA16 isolates in Anhui province were closed to the strains isolated from Yunnan, Hunan, Guangdong, Tibet and Jiangsu, especially from Hunan in 2013 and from Shenzhen of Guangdong in 2014, the identity were 96.40%-99.70%. The CVA16 strains isolated from Anhui in 2014 were all belong to genetic subtype B1b of B1 genotype was dominant, and among those isolates, several small virus transmission chains had formed with co-circulating and evolution.
China ; Enterovirus A, Human ; classification ; genetics ; isolation & purification ; Enterovirus Infections ; virology ; Genotype ; Humans ; Molecular Sequence Data ; Phylogeny ; Viral Proteins ; genetics ; metabolism

Objective To explore the clinical significance of the differential expression of PTEN in glioma tissue.Methods The mRNA and protein expressions of PTEN were assayed by reverse transcription polymerase chain reaction(RT-PCR) and immunohistochemistry in 75 human brain glioma cases.Results The positive expression rate mRNA of PTEN differed significantly between brain glioma tissue and normal brain tissue(?2=22.66,P

Objective To investigate clinical effect and safety difference of vinorelbine and docetaxel sepa-rately combined with lobaplatin in treatment of recurrent and metastatic breast cancer.Methods 160 patients with recurrent and metastatic breast cancer were chosen from July 2009 to July 2012 in our hospital,and they were random-ly divided into two groups,including A group (80 patients)with vinorelbine and B group (80 patients)with docetaxel on the basis of lobaplatin.The clinical efficacy for short -term,survival rate with follow -up and side effects incidence of both groups were compared.Results The RR and DCR of A group were 51.25%,68.75%,which of B group were 55.00%,71.25%.There was no significant difference in the clinical efficacy for short -term between the two groups (χ2 =1.04,2.37,all P >0.05).The survival rates in 1,2 and 3 years after treatment of A group were 60.00%,53.75%,28.75%,those of B group were 67.50%,57.50%,31.25%.There was no significant difference in the survival rate with follow -up between the two groups (χ2 =2.14,3.01,1.87,all P >0.05).The incidence rate of drug side effects of B group was significantly lower than A group(χ2 =13.14,9.33,15.74,11.65,8.29,all P <0.05).Conclusion Vinorelbine and docetaxel separately combined with lobaplatin in treatment of recurrent and metastatic breast cancer can efficiently control the disease progress and prolong the survival time;but docetaxel com-bined with lobaplatin treatment is helpful to reduce the risk of serious side effects.

Objective To investigate the effeets of insulin on the apoptosis of vascular endothelial cells cuinduced by burn$eruln in order to explore its possible mechamsm.Method Cultured human ECV304 cells were randomly divided into 33x)ups:control group,the ECV304 cells hured by 15%(V/V)rat normal,qertlm(t=6);bum semm group,the ECV3()4 cells simulated by 15%(V/V)self-made burn semm collected from rats with 30%TBSA full-thickness burns on the,back(n=6);and burn Serum+insulin group.the ECV304 cells cultared by insulin(10-7mol/L)and 15%(V/V) seIf-made rat bum serum(n=6).The transferase mediated nick end labeling(TUNEL) method was employed to measure the apoptosis of endothelial cells at 6 hours after stimulation.Meanwhile.immanohistochemical technique and Western blotting were used to determine the protein expressions of bcl-2 and eNOS.Data are expressed ills mean ±SEM.Statistical comparison was made using oneway analysis of vtriance.Significance was accepted at P<0.05.Results Compswith the control group,bum$erunl induced the apoptusis(18.5±3.1%)and down-regalated bcl·2(O.36±0.12)and p-eNOS(O.55±0.28)protein expressions of HUVECs(P<0.01).Burn 9AJ'unl+insulin significantly decreased the apoptosis(9.6 4-2.8%)and up-regulated bcl-2(0.944-0.25)and p-eNOS(0.89±0.16)protein expressions ofHU-VECs in comparison with the bum serllm group(P<0.01).eNOS showed no significant differences in three groups.Conclusions Insulin could markedly inhibit the apoptosis and up-regalate bcl-2 protein expression of HUVECs induced by bum serum,and its mec,harfism might involve the protein expression ofphosphorylated eNOS.

Objective To display space occupying lesions of spine,mediastina and lungs behind heart and small lesions in bilateral pulmonary fields in the same chest film without changing the exposure doses which were now used for routine chest images.Methods The distribution of current intensifying screens' intensifying materials were changed and reasonable technical processes were given.High-velocity and middle-velocity intensifying materials were placed in center and in two sides of the screen respectively.Results Tests in seven volunteers using the new kind intensifying screens demonstrated that diseases could be better shown than normal intensifying screens in chest images.Conclusion The renovation can provide more and clearer and richer stratifications of image information to help chest X-ray diagnoses.

ObjectiveTo monitor the early blood lactic acid concentration of patients with severe trauma who have been experienced routine surgery or damage control surgery,and explore the influence of surgical methods for the early lactate clearance rate.MethodsSelected 40 patients with severe trauma,they were divided into two groups with 20 cases each in accordance with the adopted operation mode,reference group by damage control surgery,and control group by routine surgery.Recorded acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ ) in patients after admission,blood lactic acid concentration at 6 h after admission and admission,calculated the early lactate clearance rate.ResultsIn reference group,blood lactic acid concentration at 6 h after admission was significantly lower than that in control group[ (3.5 ± 1.1 )mmol/L vs.(4.2 ± 1.4) mmol/L,P＜ 0.05 ],early lactate clearance rate was higher than that in control group [ (24.6 ± 6.3 )％ vs.( 11.4 ± 5.3 )％,P＜ 0.05 ].Conclusions Damage control surgery in patients with severe trauma in favour of the early removal of lactic acid,maintaining the homeostasis of the organism,is the key to improve the achievement ratio of treatment with severe trauma.

OBJECTIVETo investigate the glycolytic phenotype of SHG44 human glioma cells under hypoxic conditions and the association between cell proliferation and apoptosis and the metabolic status.

METHODSAn in vitro hypoxic cell model was established in SHG44 cells using CoCl2. Real-time PCR and Western blotting were used to assess the expressions of hypoxia-inducible factor-1α (HIF-1α) and the enzymes involved in glycolysis including PDK1, PKM2, and LDHA. Intracellular ATP levels were measured by bioluminescence assay to assess the energy metabolic status of SHG44 cells. The viability and apoptosis of the cells were examined using MTT assay and flow cytometry, respectively.

RESULTSThe cells in hypoxic culture showed obviously increased expressions of HIF-1α, LDHA, PDK1, and PKM2 at both the mRNA and protein levels as compared to those in normal cell culture. Hypoxia of the cells also resulted in a lowered cell proliferative activity and an increased apoptosis rate with lowered intracellular ATP concentrations and elevated mitochondrial membrane potential.

CONCLUSIONHypoxia can induce a glycolytic phenotype of tumor cells. The sensitivity of tumor cells to hypoxia-induced cell death is directly correlated with their metabolic status.

Adenosine Triphosphate ; metabolism ; Apoptosis ; Carrier Proteins ; metabolism ; Cell Hypoxia ; Cell Line, Tumor ; Cell Proliferation ; Central Nervous System Neoplasms ; metabolism ; pathology ; Glioma ; metabolism ; pathology ; Glycolysis ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Membrane Potential, Mitochondrial ; Membrane Proteins ; metabolism ; Protein-Serine-Threonine Kinases ; metabolism ; Thyroid Hormones ; metabolism

China ; Humans ; Streptococcus pneumoniae

Objective The aim of this study was to investigate the expression of cell cycle checkpoint kinase 1(Chk1)gene in glioblastoma cells( GBM) and its correlation with GBM cell proliferation,tumorigenic activity and prognosis. Methods The ex-pression of Chk1 in GBM cells was selected and analyzed by TCGA database and brain tumor molecular database( Rembrandt),and the level of Chk1 expression in GBM cells was detected by molecular biology techniques such as Western blot and Real-Time PCR. The expression of Chk1 was silenced by siRNA to investigate its effect on proliferation and colony-forming ability of GBM cells. The prognosis survival of GBM patients accompanying with Chk1 expression was analyzed by immunohistochemical staining and Rembrandt database. Results The results of TCGA database and Rembrandt showed that Chk1 gene was highly expressed in GBM tissues. West-ern blot and Real-Time PCR also showed that Chk1 gene was highly expressed in GBM cells. Lentiviral transfection siRNA-specific silencing of Chk1 significantly inhibited proliferation and colony-forming ability of U87 cells( P<0. 01 and P<0. 05). Prognostic survival analysis showed that GBM patients with low expression of Chk1 gene had a significantly better clinical outcome than those of GBM patients with high expression of Chk1 gene(P<0. 001). Conclusion Chk1 gene is overexpressed in GBM cells,up-regula-tion of Chk1 gene expression can promote the growth and proliferation of GBM cells,and Chk1 gene is associated with poor prognosis in GBM patients.

Objective To investigate the effect and mechanism of miR-20a-5p on human nephroblastoma cell line WiT49 transplanted tumor in nude mice.Methods The gene expression chip was downloaded from GEO database,and the differential gene miR-20a-5p was obtained by GEO2R.The NF-κB gene was positively correlated with the expression of miR-20a-5p through cBioPortal database.The target gene of miR-20a-5p was predicted to be NFKBIB of the NF-κB transcription factor suppressor protein family by targetscan database,and was verified by dual luciferase assay.Nephroblastoma cell line WiT49 was cultured in vitro and transfected into WiT49 cells with lentiviral vectors constructed with miR-20a-5p mimics and its suppressor gene.Twelve nude mice were randomly divided into three groups:WiT49 model group,WIT49-miR-20a-5p overexpression group and WIT49-miR-20a-5p knockdown group.The tumor mass and volume of each group were detected by tumor formation experiment in nude mice.real time fluorescent quantitative polymerase chain reaction(qRT-PCR)was used to detect the expression of miR-20a-5p,NFKBIB and NF-κB in each group;CCK-8 cell proliferation assay was used to verify the proliferation of tumor cells in each group.Results miR-20a-5p is highly expressed in nephroblastoma and is positively correlated with the expression of NF-κB.miR-20a-5p and NFKBIB have mutual binding sites and binding effects.In the tumor formation experiment of nude mice,the tumor volume and mass of WIT49-miR-20a-5P overexpression group were significantly increased compared with WiT49 model group,and the difference was statistically significant(P<0.05).In the qRT-PCR test,the expressions of miR-20a-5p and NF-κB in the WIT49-miR-20a-5p overexpression group were higher than those in the WiT49 model group,and NFKBIB expression in the WIT49-miR-20a-5p overexpression group was lower than that in the WiT49 model group,with statistical significance(P<0.05).CCK-8 cell proliferation assay showed that the absorbance of WIT49-miR-20a-5p overexpression group at 24 and 48 hours was higher than that of WiT49 model group,and the absorbance of WIT49-miR-20a-5p knockdown group at 24,48 and 72 hours was lower than that of WiT49 model group,and the difference was statistically significant(P<0.05).Conclusion miR-20a-5p may promote the growth of human nephroblastoma cell WiT49 transplanted tumor in nude mice by regulating NFKBIB activation of NF-κB pathway.