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.

Polymorphisms in DNA repair genes may alter DNA repair capacity and, consequently, lead to genetic instability and carcinogenesis. Several studies have investigated the association of the Asp312Asn and Lys751Gln polymorphisms in the xeroderma pigmentosum complementation group D (XPD) gene with the risk of non-Hodgkin’s lymphoma (NHL), but the conclusions have been inconsistent. Therefore, we performed this meta-analysis to more precisely estimate these relationships. A systematic literature search was performed using the PubMed, Embase, and Chinese Biomedical (CBM) databases. Ultimately, 6 studies of Asp312Asn, comprising 3,095 cases and 3,306 controls, and 7 studies of Lys751Gln, consisting of 3,249 cases and 3,676 controls, were included. Pooled odds ratios (ORs) and 95%confidence intervals (CIs) were calculated to assess the strength of each association. Overal , no association was observed between the Asp312Asn polymorphism and NHL risk (homozygous:OR=1.11, 95%CI=0.94-1.32;heterozygous:OR=1.00, 95%CI=0.89-1.11;recessive:OR=1.12, 95%CI=0.95-1.31;dominant:OR=1.02, 95%CI=0.92-1.13;and al ele comparison:OR=1.04, 95%CI=0.96-1.12) or between the Lys751Gln polymorphism and NHL risk (homozygous:OR=0.97, 95%CI=0.83-1.15;heterozygous:OR=0.96, 95%CI=0.86-1.06;recessive:OR=1.00, 95%CI=0.86-1.16;dominant:OR=0.96, 95%CI=0.87-1.06;and al ele comparison:OR=0.98, 95%CI=0.91-1.05). Furthermore, subgroup analyses did not reveal any association between these polymorphisms and ethnicity, the source of the controls, or the NHL subtype. These results indicated that neither the Asp312Asn nor Lys751Gln XPD polymorphism was related to NHL risk. Large and well-designed prospective studies are required to confirm this finding.

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

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

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

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.

Objective To investigate the resistance of clinical Stenotrophomonas maltophilia isolates from 15 hospitals in several regions of China during 2011.Methods Fifteen repre-sentative general hospitals were involved in this program. Bacterial susceptibility testing was carried out by means of a unified protocol using Kirby-Bauer method and MIC determi-nation.Results were analyzed according to CLSI 2011 break-points.Results Majority (93.3%) of the 1 889 clinical strains of S.maltophilia were isolated from inpatients.On-ly 6.7% of the isolates were from outpatients.About 62.9% of these S .maltophilia strains were isolated from old patients whose age was 60 years or older.Only 8.2% of the strains were from the patients younger than 18 years old.Sputum and re-spiratory tract secretion were the most common specimen source,accounting for 82.6%.Another 4.2% isolates were from blood,abdominal fluid and other sterile body fluids.The percentage of the S .maltophilia strain resistant to trimethoprim-sul-famethoxazole,levofloxacin and minocycline was 16.6%,10.0% and 1.8%,respectively.The strains resistant to cefopera-zone-sulbactam accounted for 19.0%.About 37.1% of the strains isolated from blood or sterile body fluids were resistant to trimethoprim-sulfamethoxazole,significantly higher than the strains from urine or wound specimens (P < 0.01).Conclusions S.maltophilia strains are mainly isolated from inpatients.The most common source is sputum and other respiratory speci-mens.Most of the patients with S.maltophilia isolate are 60 years of age or older.The S.maltophilia strains are constitu-tively resistant to several antibacterial agents,but showed relatively lower resistance to trimethoprim-sulfamethoxazole,levo-floxacin and minocycline.Cefoperazone-sulbactam is still active against these strains.The antimicrobial therapy targeting S. maltophilia infections should be selected cautiously according to the results of antimicrobial resistance surveillance.

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 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 .