Objective To build the experimental basement for the clinical use of cytokines induced killer(CIK)cells from the cord blood mononuclear cells(CBMNC)in tumor adoptive cellular immunotherapy, an effective protocol for their proliferation in vitro and cytotoxicity of CIK cells was established.Methods The lymphocytes from umbilical cord blood were isolated by density gradient centrifugation and suspended in medium with CD_3 mAb,rIL-2,rIL-1 and IFN-? as inducing agents to prepare CIK cells.At the same time, the lymphokine activated killer(LAK)and CBMNC were set as controls,which were only added IL-2 and not any cytokines during the whole culture.The changes of CIK cells before and after induction were observed with microscope and the phenotypes of the cells were analyzed by using flow cytometry.The proliferation of CIK cells were determined by trypan blue exclusion assay and the cytotoxic activity to lung cancer cell were tested with MTF method.Results According to the experiment,combining use of four types of cytokines could generate a great deal of CIK cells possessing highly cytotoxicity.From day 5 CIK cells became to prolif- erate and reached the peak at day 14.During the whole period,the relative percentage of CD_3~+ CD_(56)~+ cells in- creased significantly.Compared with LAK cells,which reached the proliferation peak at day 7 and then showed no evident proliferation.The control cells(CBMNC)showed no evident change of phenotypes and proliferation.CIK cells showed a higher antitumor activity on the tumor cells than LAK cells and CBMNC in vitro.Conclusion Umbilical cord blood can generate a great deal of CIK cells combining used with cy- tokines.Compared with classic LAK cells,umbilical cord blood CIK cells have the advantages of rapid prolif- eration speed and powerful cytotoxicity.CIK cells will be promising as a new strategy for the adoptive cellular immunotherapy of tumor.

OBJECTIVETo explore the correlations of circulating tumor cells (CTCs) and disseminated tumor cells (DTCs) with the clinicopathological characteristics, prognostic events, and survival outcomes in esophageal cancer (EC) patients.

METHODSThe PubMed, Web of Science, Embase database and Cochrane database were searched for studies reporting the outcomes of interest. The studies were selected according to established inclusion/exclusion criteria. Meta-analysis of the studies was performed using Review Manager 5.3 and Stata12.0 software with the odds ratio (OR), risk ratio (RR) , hazard ratio (HR) , and 95% confidence interval (95% CI) as the effect indexes.

RESULTSNineteen studies involving a total of 1766 patients were included in the analysis. Significant correlations of CTCs and DTCs were found with the clinicopathological parameters including the tumor stage (OR=1.95), depth of invasion (OR=1.99), lymph node metastasis (OR=2.44), distal metastasis (OR=5.98), histological differentiation (OR=1.67) and lymphovascular invasion (OR=4.48). CTCs and DTCs were also correlated with the prognostic events including relapse (RR=6.86) and metastasis (RR=3.22) and with the survival outcomes including the overall survival (OS) overall analysis (HR=3.46) and disease-free survival/progression-free survival (DFS/PFS) overall analysis (HR=3.00).

CONCLUSIONCTCs and DTCs are significantly associated with an advanced tumor stage, depth of tumor invasion, lymph node metastasis, distant metastasis before therapy, differentiation, lymphovascular invasion, relapse and metastasis in patients with EC. They are also significantly correlated with a poorer survival for OS and DFS/PFS to serve as clinical and prognostic predictors in patients with EC.

Disease-Free Survival ; Esophageal Neoplasms ; diagnosis ; Humans ; Lymphatic Metastasis ; Neoplasm Recurrence, Local ; Neoplastic Cells, Circulating ; Odds Ratio ; Prognosis ; Proportional Hazards Models ; Survival Analysis

This study was purposed to construct a vector containing human suppressor gene p53 and p16, and to investigate their expression and effect on K562 and HL-60 cells. pBudCE4.1-53-16 is a vector designed for simultaneous expression of human suppressor gene p53 and p16 in mammalian cell line. After transfection into K562 cells with lipofectamine(TM) 2000, the expression of p53 and p16 genes was detected by Western blot and immunocytochemical method. The growth curve, apoptosis, cell cycle were assayed by CCK-8 and flow cytometry. The results showed that the recombinant plasmid pBudCE4.1-53-16 was constructed successfully and were verified by PCR and restriction analysis. The expression of P53 and P16 protein could be detected after transfection into leukemia cells (K562 and HL-60) for 48 hours. As compared with control group, the cell proliferation in experimental group was inhibited, the cells were arrested in G0 phase and apoptotic cells increased (p<0.001). It is concluded that the recombinant plasmid pBudCE4.1-53-16 has been established. p16 and p53 in the recombinant plasmid pBudCE4.1-53-16 synchronously express in leukemic cells after transfection in vitro for 2 days and results in reduced proliferation, G0 arrest and apoptosis increase.
Apoptosis ; genetics ; Cell Cycle ; genetics ; Cell Proliferation ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; Gene Expression ; Genes, p53 ; Genetic Vectors ; HL-60 Cells ; Humans ; K562 Cells ; Plasmids ; Transfection

OBJECTIVETo clone the human tissue factor pathway inhibitor-2 (hTFPI-2) gene and express it by using prokaryotic expression system.

METHODSThe hTFPI-2 coding region was obtained by RT-PCR from human placenta total RNA. The coding fragment was then inserted into prokaryotic expression vector pET19b and expressed in E. coli BL21 by IPTG induction. The produced inclusion bodies were dissolved by denaturalizing chemicals, purified by ion exchange chromatograph, and refolded in air to form proper disulfide bonds. Chromogenic and gelatin zymography methods were used to evaluate the inhibiting effects of hTFPI-2 on trypsin, plasmin and MMPs individually. The inhibitory activity of hTFPI-2 on fabrisarcoma was investigated by matrigel.

RESULTSThe coding fragment of hTFPI-2 was cloned successfully and the protein was expressed as inclusion bodies which account for 20% - 30% of total host protein. The refolded hTFPI-2 could inhibit the invasive ability of fibrisarcoma HT-1080 as well as activity of plasmin, trypsin and MMPs.

CONCLUSIONSThe activated hTFPI-2 is obtained by using prokaryotic expressed system effectively.

Cloning, Molecular ; Escherichia coli ; metabolism ; Gene Expression ; Glycoproteins ; biosynthesis ; genetics ; Humans ; Placenta ; cytology ; RNA ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction

Identifying risk factors of the disease are one of the main tasks of epidemiology. With the advancement of omics technologies (e.g., genome, transcriptome, proteome, metabolome, and exposome), cancer etiology research has entered the stage of systems epidemiology. Genomic research identifies cancer susceptibility loci and uncovers their biological mechanisms. Exposomic research investigates the impact of environmental factors on biological processes and disease risks. The metabolome is downstream of biological regulatory networks, reflecting the effects of the gene, environment, and their interactions, which can help elucidate the biological mechanisms of genetic and environmental risk factors and identify new biomarkers. Here, we reviewed the applications of genomic, exposomic, and metabolomic studies in the etiologic research on cancer. We summarized the importance of multi-omics approaches and systems epidemiology in cancer etiology research and outlined future perspectives.
Humans ; Multiomics ; Genomics ; Metabolomics ; Neoplasms/genetics* ; Biomarkers

Objective To study the prevalence and distribution of mental disorders among registered and non-registered residents in Shenzhen. Methods An epidemiological survey on mental disorders were carried out in Shenzhen by stratified multi-stage randomized sampling method; 7134 respondents were assessed through face-to-face interview, using the WHO standardized version on World Mental Health (WMH) Survey Initiative of the Composite International Diagnostic Interview (CIDI3.1). Results (1)The weighting prevalence of mental disorders was 21.87%. The prevalence of non-registered residents was significantly higher than that of the registered residents (22.34% vs. 19.99% ; OR= 1.15,95%CI: 1.03-1.29; P<0.05) and the prevalence of females was significantly higher than that of males (22.68% vs. 19.67%; OR=1.20,95%CI: 1.07-1.34; P<0.05). The weighting prevalence of mood disorders, anxiety disorders and psychoses were 9.62%, 14.45% and 1.40%, respectively. (2) The weighting twelve-month incidence of mental disorders was 13.42%. The incidence of non-registered residents was significantly higher than that of the registered residents (13.80% vs. 11.90%; OR=1.19, 95%CI: 1.03-1.36; P<0.05). (3)The co-morbidity rate between mental disorders was 35.76%. (4)The prevalence and severity of mental disorders were associated with sex, household situation of registration, marital status, education, economic condition and occupation status. Conclusion Mental disorders have become common diseases and serious public health problem in Shenzhen, with non-registered residents and females deserve more attention.

OBJECTIVETo investigate effects of retinol on the expressions of epidermal growth factor (EGF), stem cell factor (SCF), colony-stimulating factor 1 (CSF1) and leukemia inhibitory factor (LIF) in cultured human umbilical-derived mesenchymal stem cells (UCMSCs).

METHODSHuman UCMSCs were isolated from human umbilical cord and identified for immunophenotypes. The cells were then cultured in DMEM/F12 media supplemented with 12% fetal bovine serum (FBS), 12% FBS+1 µmol/L retinol, 15% knockout serum replacement (KSR) and 15% KSR+ 1 µmol/L retinol. The expressions of the cytokines EGF, SCF, CSF1 and LIF in the cells were detected using RT-PCR and ELISA.

RESULTSThe isolated cells exhibited characteristic immunophenotypes of human UCMSCs and expressed EGF, CSF1 and SCF at both mRNA and protein levels but not LIF protein. Retinol (1 µmol/L) significantly promoted the expressions of SCF and CSF1 at both mRNA and protein levels but did not result in changes of EGF and LIF expressions in human UCMSCs.

CONCLUSIONRetinol at the concentration of 1 µmol/L can promote expression of SCF and CSF1 in human UCMSCs in vitro.

Cell Differentiation ; Cells, Cultured ; EGF Family of Proteins ; metabolism ; Humans ; Immunophenotyping ; Leukemia Inhibitory Factor ; metabolism ; Macrophage Colony-Stimulating Factor ; metabolism ; Mesenchymal Stromal Cells ; drug effects ; metabolism ; Stem Cell Factor ; metabolism ; Umbilical Cord ; cytology ; Vitamin A ; pharmacology

Objective To investigate the susceptibility and resistance of clinical isolates from hospitals in several regions of China .Methods Fifteen general hospitals and two children′s hospitals were involved in this program . Antimicrobial susceptibility testing was carried out according to a unified protocol using Kirby‐Bauer method or automated systems .Results were analyzed according to CLSI 2014 breakpoints .Results A total of 78 955 clinical isolates were collected from January to December 2014 ,of which gram negative organisms and gram positive cocci accounted for 72 .6% and 27 .4% ,respectively . Methicillin‐resistant strains in S .aureus(MRSA)and coagulase negative Staphylococcus(MRCNS)accounted for an average of 44 .6% and 83 .0 % ,respectively .The resistance rates of methicillin‐resistant strains to β‐lactams and other antimicrobial agents were much higher than those of methicillin‐susceptible strains .However ,92 .0% of MRSA strains were still susceptible to trimethoprim‐sulfamethoxazole ,while 85 .6% of MRCNS strains were susceptible to rifampin .No staphylococcal strains were found resistant to vancomycin ,teicoplanin or linezolid .In Enterococcus spp .,the resistance rates of E .f aecalis strains to most tested drugs (except chloramphenicol) were much lower than those of E . f aecium .Some strains of both species were resistant to vancomycin .Vancomycin resistant strains of E . f aecalis and E . f aecium were mainly V anA ,V anB or V anM type based on their phenotype or genotype .Regarding non‐meningitis S .pneumoniae strains ,the prevalence of penicillin‐susceptible S .pneumoniae strains isolated from both adults and children were higher than those isolated in 2013 ,but the prevalence of penicillin‐intermediate S . pneumoniae or penicillin‐resistant S . pneumoniae strains decreased . The prevalence of ESBLs producingstrainswas55.8% in E.coliand29.9% in Klebsiellaspp.(K.pneumoniaeand K.oxytoca)and24.0% in Proteus mirabilis isolates on average . ESBLs‐producing Enterobacteriaceae strains were more resistant than non‐ESBLs‐producing strains in terms of antibiotic resistance rates . The strains of Enterobacteriaceae were still highly susceptible to carbapenems .Overall less than 10 % of these strains were resistant to carbapenems . About 62 .4% and 66 .7% of Acinetobacter spp .(A .baumannii accounts for 93 .0 % ) strains were resistant to imipenem and meropenem ,respectively . Compared with the data of year 2013 ,extensively‐drug resistant strains in K . pneumoniae and A .baumannii increased . Conclusions The antibiotic resistance of clinical bacterial isolates is growing .The disseminated multi‐drug or pan‐drug resistant strains in a special region poses a serious threat to clinical practice and implies the importance of strengthening infection control .

Objective To investigate the antimicrobial resistance of clinical strains of K lebsiella spp .isolated from 15 hospitals in China CHINET during 2012 .Methods Kirby-Bauer method and automatic microbiology analysis system were employed to study the antimicrobial resistance . WHONET 5 .6 software was applied for data analysis according to Clinical and Laboratory Standards Institute (CLSI) 2012 breakpoints .Results A total of 9 621 clinical K lebsiella isolates were analyzed ,including 8 772 strains of K . pneumoniae and 804 strains of K . oxytoca . About 54 .9% (5 285/9 621) of the K lebsiella strains were isolated from sputum ,and 16 .3% (1 564/9 621) were isolated from pediatric patients .Antimicrobial susceptibility testing showed that about 8 .9% ,10 .8% and 12 .9% of the strains were resistant to imipenem ,meropenem and ertapenem ,respectively .About 14 .1% and 17 .0% of the strains were resistant to piperacillin-tazobactam and cefoperazone-sulbactam , respectively . Carbapenem-resistant K lebsiella strains were identified from all the 15 hospitals ,including 945 strains of K .pneumoniae and 45 strains of K .oxytoca ,which were resistant to either imipenem ,meropenem or ertapenem .Conclusions The Klebsiella isolates collected from 15 hospitals in China during 2012 are relatively sensitive to carbapenems ,cefoperazone-sulbactam and piperacillin-tazobactam .The prevalence of carbapenem-resistant strains is still increasing in China ,about 10 .3% in 2012 ,and relatively higher in Eastern China .More efforts should be made to control the superbug .

Objective To investigate the antimicrobial resistance in the A cinetobacter baumannii strains in different parts of China during 2012 .Methods A total of 8 739 clinical isolates of Acinetobacter were collected from 13 general hospitals and two children’s hospitals ,of which most were A . baumannii (89 .6% , 7 827/8 739 ) . Antimicrobial susceptibility testing was carried out by means of Kirby-Bauer method according to the unified protocol . The susceptibility testing data were analyzed by WHONET 5 .6 software according to CLSI 2013 breakpoints .Results Majority (85 .4% ) of the Acinetobacter strains were isolated from inpatients .The remaining 14 .6% were from outpatients and emergency room patients .Of the 7 827 strains of A .baumannii , 10 .9% ,35 .2% ,35 .7% and 43 .4% were resistant to tigecycline ,minocycline ,cefoperazone-sulbactam and amikacin , respectively .The percentage of A .baumannii resistant to imipenem and meropenem was 63 .5% and 68 .2% ,respectively . The antimicrobial resistant pattern varied in different hospitals . The resistance of A . baumannii varied between different clinical departments .A number of pandrug resistant (PDR) (20 .0% ,1 567/7 827) and multidrug-resistant (MDR) (45 .0% , 3 521/7 827 ) A . baumannii were identified . Conclusions A . baumannii is the most popular pathogenic bacteria among Acinetobacter .The antibiotic resistance of A .baumannii is still increasing .Cefoperazone-sulbactam and minocycline has good in vitro antibacterial activity against A .baumannii .The antibiotic resistance of A .baumannii varies greatly with hospital and department .