谷氨酸棒状杆菌在乙酸盐和葡萄糖基质上的蓝白生长筛选(英文)

在谷氨酸棒状杆菌乙酸盐代谢调控关键酶—异柠檬酸裂解酶 ace A基因两段启动子区域后面组装了无启动子的β-半乳糖苷酶报道基因 ,并且带上四环素抗性标记基因构建了两个检测质粒 p ROH7和 p ROHO.通过该两个质粒转化了谷氨酸棒状杆菌野生型菌株 Corynebacterium glutamicum ACTT14 75 2 ,分别获得了含有两个检测质粒的两种谷氨酸棒状杆菌 ,由此建立了谷氨酸棒状杆菌在乙酸盐和葡萄糖基质上的两个蓝白生长筛选系统 .结果显示当两种菌株生长在含有葡萄糖的基本培养基上时呈现白色菌落 ,而生长在含有乙酸盐的基本培养基上时呈现蓝色菌落 ;其中菌落颜色还随菌株中检测质粒p ROH7和 p ROHO的不同而呈现淡蓝色和深蓝色的变化 .含有检测质粒 p ROH7和 p ROHO菌落的颜色变化反映了细胞在葡萄糖和乙酸盐不同和碳源上的调控状态 。

Isolation and characterization of atrazine-degrading Arthrobacter sp. AD26 and use of this strain in bioremediation of contaminated soil

A bacterial strain (AD26) capable of utilizing atrazine as a sole nitrogen source for growth was isolated from an industrial wastewater sample by enrichment culture. The 16S rRNA gene sequencing identified AD26 as an Arthrobacter sp. PCR assays indicated that AD26 contained atrazine-degrading genes trzN and atzBC. The trzN gene of AD26 only differs from the trzN of Arthrobacter aurescens TC1 by one base (A→T at 907) and one amino acid (Met→Leu at 303). The specific activity of trzN of AD26 in crude cell ext...

Cloning and expression of NS3 cDNA fragment of HCV genome of Hebei isolate in E.coli

AIM To obtain greater antigenicity of HCV NS3 protein. METHODS The HCV NS3 cDNA fragment was amplified by reverse transcription polymerase chain reaction from the sera of the HCV infected patients. The DNA sequence was determined by dideoxy mediated chain termination method using T7 polymerase. HCV NS3 protein was expressed in E. coli . RESULTS Sequence analysis indicated that the HCV isolate of this study belongs to HCV Ⅱ; SDS PAGE demonstrated an M r 23800 and an M r 22000 recombinant protein band which amount to 14% and 11% of the total bacterial proteins separately. Western blotting and ELISA showed NS3 protein possessed greater antigenicity. CONCLUSION Recombinant HCV NS3 protein was expressed successfully, which provided the basis for developing HCV diagnostic reagents.

Adhesion and immunomodulatory effects of Bifidobacterium lactis HN019 on intestinal epithelial cells INT-407

AIM:To elucidate the adherence and immunomodulatory properties of a probiotic strain Bifidobacterium lactis(B.lactis) HN019.METHODS:Adhesion assays of B.lactis HN019 and Salmonella typhimurium(S.typhimurium) ATCC 14028 to INT-407 cells were carried out by detecting copies of species-specific genes with real-time polymerase chain reaction.Morphological study was further conducted by transmission electron microscopy.Interleukin-1β(IL-1β),interleukin-8,and tumor necrosis factor-α(TNF-α) gene expression were assessed while enzyme linked immunosorbent assay was used to detect IL-8 protein secretion.RESULTS:The attachment of S.typhimurium ATCC 14028 to INT407 intestinal epithelial cells was inhibited significantly by B.lactis HN019.B.lactis HN019 could be internalized into the INT-407 cells and attenuated IL-8 mRNA level at both baseline and S.typhimuriuminduced pro-inflammatory responses.IL-8 secretion was reduced while IL-1β and TNF-α mRNA expression level remained unchanged at baseline after treated with B.lactis HN019.CONCLUSION:B.lactis HN019 does not up-regulate the intestinal epithelium expressed pro-inflammatory cytokine,it showed the potential to protect enterocytes from an acute inflammatory response induced by enteropathogen.

Effect of S1P5 on proliferation and migration of human esophageal cancer cells

AIM:To investigate the sphingosine 1phosphate (S1P) receptor expression profile in human esophageal cancer cells and the effects of S1P5 on proliferation and migration of human esophageal cancer cells. METHODS: S1P receptor expression profile in human esophageal squamous cell carcinoma cell line Eca109 was detected by semiquantitative reverse trans cription polymerase chain reaction. Eca109 cells were stably transfected with S1P5EGFP or controlEGFP constructs. The relation between the responses of cell proliferationand migration to S1P and S1P5 expres sion was evaluated by 3(4,5dimethylthiazol2yl)2,5diphenyl tetrazolium bromide and migration assay, respectively. RESULTS: Both normal human esophageal mucosal epithelium and Eca109 cells expressed S1P1, S1P2, S1P3 and S1P5, respectively. Esophageal mucosal epithelium expressed S1P5 at a higher level than Eca109 cell line. S1P5 overexpressing Eca109 cells displayed spindle cell morphology with elongated and extended filopodialike projections. The proliferation response of S1P5transfected Eca109 cells was lower than that of control vectortransfected cells with or without S1P stimulation (P < 0.05 or 0.01). S1P significantly inhibited the migration of S1P5transfected Eca109 cells (P < 0.001). However, without S1P in transwell lower chamber, the number of migrated S1P5transfected Eca109 cells was greater than that of control vectortransfected Eca109 cells (P < 0.001).CONCLUSION: S1P binding to S1P5 inhibits the proliferation and migration of S1P5transfected Eca109 cells. Esophageal cancer cells may downregulate the expression of S1P5 to escape the inhibitory effect.

Selective blockade of NF-kappa B by novel mutated I kappa B alpha suppresses CD3/CD28-induced activation of memory CD4(+) T cells in asthma

Background:Nuclear factor kappa B(NF-kappa B) overactivation plays a crucial role in T-helper 2 (Th2)-biased allergic airway inflammation by increased activation and decreased apoptosis of CD4(+) T cells. We have shown that targeted NF-kappa B suppression in dendritic cells by adenoviral gene transfer of a novel mutated inhibitor of NF-kappa B(I kappa B alpha) (AdI kappa B alpha M) contributes to T-cell tolerance, but the immunosuppressive action of AdI kappa B alpha M on memory(CD45RO(+)) CD4(+) T cells remains enigmatic. Methods:CD45RO(+) T cells from Dermatophagoides farinaei-sensitized asthmatic patients, untransfected or transfected with AdI kappa B alpha M or AdLacZ(beta-galactosidase) for 24 h, were stimulated with anti-CD3(1.0 mu g/ml) plus anti-CD28(0.5 mu g/ml) monoclonal antibody for an additional 24 h. I kappa B alpha M transgene expression and NF-kappa B activation were detected by polymerase chain reaction(PCR), reverse transcription-PCR(RT-PCR), Western blot analysis, and electrophoretic mobility shift assay. Phenotype and apoptosis were measured by flow cytometry, annexin V binding, and terminal deoxyribonucleotidyl transferase-mediated dUTP nick end labeling analyses. Cytokine production and cell proliferation were determined using enzyme-linked immunosorbent assay andH-3 thymidine incorporation. Results:A unique 801-bp I kappa B alpha M cDNA and a dose-dependent increase in I kappa B alpha M transgene expression were observed in AdI kappa B alpha M-transfected CD45RO(+) T cells. Significantly, AdI kappa B alpha M inhibited CD3/CD28-mediated NF-kappa B activation in CD45RO(+) T cells, leading to evident apoptosis, reduction of eotaxin, RANTES, Th1 interferon (IFN)-gamma and interleukin (IL)-2, and Th2 (IL-4, IL-5, and IL-13 despite a slight decrease in IL-10) cytokines and secondary proliferative response. AdI kappa B alpha M also upregulated cytotoxic T lymphocyte-associated antigen 4(CTLA-4) and downregulated CD69 besides no change in CD28. Conclusion:I kappa B alpha M might be beneficial to augment memory CD4(+) T-cell tolerance through modulating B7-CD28/CTLA-4 co-stimulatory pathways and NF-kappa B-dependent cytokine profiles in allergic inflammatory diseases including asthma.

转IE2基因荷瘤小鼠中ATF5功能影响的研究

利用转IE2基因小鼠荷瘤的方法,对肿瘤组织中ATF5转录激活因子进行定性和定量的检测,从而探究巨细胞病毒(HCMV)即刻早期蛋白IE86对ATF5功能的影响。通过鼠胶质瘤细胞GL261对转基因鼠进行荷瘤,采用RT-PCR和Western blot的方法,对荷瘤鼠的肿瘤组织中转录因子ATF5的表达量进行检测。采用RT-PCR和Western blot的方法检测荷瘤鼠肿瘤组织中转录因子ATF5的表达,结果显示,转IE2基因小鼠体内ATF5表达量较正常小鼠ATF5表达量高。ATF5在转IE2基因荷瘤鼠中呈现高表达状态,ATF5是一个与凋亡密切相关的转录因子,在凋亡、分化及发育等生理过程中发挥着重要作用。

Cyclosporin A protects Balb/c mice from liver damage induced by superantigen SEB and D-GaIN

AIM To investigate the pathogenic effect ofSEB and D-GalN on liver and the protection ofcyclosporin A, the relationship between hepaticapoptosis and necrosis and the possiblemechanism of acute hepatic necrosis.METHODS After staphylococcal enterotoxin B(SEB ) mixed with D--galactosamine (D-GaiN )were injected intraperitoneally into Balb/c miceand those previously treated with cyclosporin A,blood samples were collected and livers wereisolated at 2, 6, 12 and 24 h. Patterns othepatocellular death were studiedmorphologically and biochemically, circulatingcytokines (TNF-a, IFN--y ) and mice mortalitywithin 24h was assessed.RESU’LTS The SEB could induce the typicalapoptotic changes of hepatocytes, the D-GaiNcould induce hepatocytes apoptosis anddegeneration at the same time, and the micehaving received the SEB + D-GaiN injectionsdeveloped apoptosis at 2 and 6 h, but after 12 hhepatocytes were characterized by severein jury, whereas all the examinations in thecyclosporin A treated mice were normal.CONCLUSION Hepatic cell apoptosis might berelated to necrosis, and massive hepatocyteapoptosis is likely the initiating step of acutehepatic necrosis in mice. The effects induced bySEB and D--GaiN on hepatocytes might bemediated by T cells, and could be prevented bycyclosporin A.

LOH analysis of genes around D4S2964 identifies ARD1B as a prognostic predictor of hepatocellular carcinoma

AIM:To investigate genes around the locus D4S2964 affected by loss of heterozygosity(LOH) and their clinical implications.METHODS:Four hundred and forty single nucleotide polymorphisms(SNPs) located at 49 genes around D4S2964 were selected from the National Center for Biotechnology Information website for the SNPs microarray fabrication.LOH of SNPs markers in 112 cases of hepatocellular carcinoma(HCC) tissues and paired adjacent liver tissues were investigated by the SNPs microarray.The correlation between allelic losses with clinicopathological features and overall survival was analyzed.RESULTS:A f ine map of LOH of SNPs in genes around D4S2964 was plotted.The average frequency of LOH in genes was 0.39.A correlation between cirrhosis and the FAL index(fractional allelic loss) was found(P = 0.0202).Larger tumor size was found to be signif icantly associated with LOH in genes ADP-ribosyltransferase 3(ART3),nucleoporin 54 kDa(NUP54),scavenger receptor class B,member 2(SCARB2) and coiled-coil domain containing 158(CCDC158)(P = 0.043,P = 0.019,P = 0.001,P = 0.037,respectively).Kaplan-Meier analysis showed that patients with LOH in ARD1 homolog B(ARD1B) and septin 11(SEPT11) had a significantly lower survival rate than those with retention(P = 0.021 and P = 0.004,respectively).A Cox regression model suggested that LOH in ARD1B and SEPT11,respectively,were predictors of the overall survival in HCC(P = 0.006 and P = 0.026,respectively).CONCLUSION:LOH in genes around D4S2964 may play an important role in HCC development and progression.LOH in ARD1B and SEPT11 could serve as novel prognostic predictors in HCC patients.

IFN-α治疗后抗HBs阳性的慢性乙肝患儿血清HBV-DNA仍可能持续存在,与其产生的特异性IgG亚类有关

Background/Aims: We examined the persistence of HBV-DNA in sera collected 4- 10 years after IFN-αtherapy from patients with chronic hepatitis B who had ser oconverted to anti-HBs antibodies. We also wanted to assess whether any associa tion exists between HBV-DNA status and the IgG anti-HBs subclass responses. Me thods: Sera were obtained from 38 patients and the following parameters were det ermined in each of them: (1) serological markers of HBV; (2) concentrations of I gM, IgG, IgA; (3) total IgG subclasses. HBV-DNA and IgG anti-HBs subclasses we re determined in anti-HBs positive sera.Results: Four to 10 years after therapy , anti-HBs were found in 37 of 38 patients (GMT: 775 IU/L). In 13 of them (35.1 %) free and/or bound HBV-DNA was present in sera. Significant differences in t he profiles of IgG anti-HBswere observed when the HBV-DNA status was considered. Patients with undetectable HBV-DNA responded mainly with IgG1 and/or IgG3, while in the HBV-DNA-positive group, a high contribution of IgG4 was found. Conclusions: Our study showed that HBV-DNA may persist for a long t ime after IFN-αtherapy despite the appearance of anti-HBs antibodies. The mon itoring of specific IgG subclasses may be of predictive value for HBV-DNA persi stence.

Nosocomial bloodstream infection in patients caused by Staphylococcus aureus： drug susceptibility, outcome, and risk factors for hospital mortality

Background Previous studies have different viewpoints about the clinical impact of methicillin resistance on mortality of hospital-acquired bloodstream infection （BSI） patients with Staphylococcus aureus （S.aureus）.The objective of this study was to investigate the mortality of hospital-acquired BSI with S.aureus in a military hospital and analyze the risk factors for the hospital mortality.Methods A retrospective cohort study was performed in patients admitted to the biggest military tertiary teaching hospital in China between January 2006 and May 2011.All included patients had clinically significant nosocomial BSI with S.aureus.Multivariate Logistic regression analysis was used to identify the risk factors for hospital mortality of patients with S.aureus BSI.Results One hundred and eighteen patients of more than one year old were identified as clinically and microbiologically confirmed nosocomial bacteraemia due to S.aureus,and 75 out of 118 patients were infected with methicillin-resistant S.aureus （MRSA）.The overall mortality of nosocomial S.aureus BSI was 28.0%.Methicillin resistance in S.aureus bacteremia was associated with significant increase in the length of hospitalization and high proportion of inappropriate empirical antibiotic treatment.After Logistic regression analysis,the severity of clinical manifestations （APACHE Ⅱ score） （odds ratio （OR） 1.22,95% confidence interval （CI） 1.12-1.34） and inadequacy of empirical antimicrobial therapy （OR 0.25,95% CI 0.09-0.69） remained as risk factors for hospital mortality.Conclusions Nosocomial S.aureus BSI was associated with high in-hospital mortality.Methicillin resistance in S.aureus has no significant impact on the outcome of patients with staphylococcal bacteremia.Proper empirical antimicrobial therapy is very important to the prognosis.

The chimeric Japanese encephalitis/Dengue 2 virus protects mice from challenge by both dengue virus and JEV virulent virus

Dengue fever （DF） is the most significant emerging arbovirus disease, which is caused by four distinct serotypes of Dengue viruses （DENY-1 to DENV-4）. Serotype-specific immunity does not confer cross-protection against secondary infec- tions from heterologous serotypes, which leads to dengue haemorrhagic fever （DHF） and dengue shock syndrome （DSS） as a result of antibody-dependent enhancement （ADE） of infection that is mediated by non-neutralizing, cross-reac- tive antibodies （Kliks et al., 1989）. The intense efforts to develop a safe and efficient DENV vaccine have involved using a variety of traditional biological technologies in the past 50 years （Durbin et al., 2005; McArthur et al., 2008）, yet no commercial DENV vaccine has been generated until now.

Identification of endoplasmic reticulumshaping proteins in Plasmodium parasites

Dear Editor, In eukaryotic cells, the endoplasmic reticulum （ER） is a continuous membrane system involved in many cdtical cellular processes, including protein synthesis, lipid synthesis, and calcium storage. Morphologically, the ER is composed of cistern-like sheet structures and a reticular network of tubules.

Etiological and molecular-epidemiological analysis on enterovirus associated encephalitis in Zhejiang,2008-2012

Objective In order to investigate etiology and molecular-epidemiological characteristics of enterovirus associated encephalitis(EAE)in Zhejiang,2008—2012.Methods Cerebrospinal fluid and stool specimens were collected from suspected EAE patients,who were admitted to our hospitals.RD and Hep-2 cell lines were used to

Anti-tumor effect of Sendai virus Tianjin strain defective interfering particles on tumor-bearing mice

Objective To explore the anti-tumor effect and its mechanism of Sendai virus Tianjing strain defective interfering particles(DIP)on mouse models of colon carcinoma.Methods CT26 cells(5×106/0.1 ml)were subcutaneously injected into the back of Bal B/c mice to establish murine colon carcinoma model.After the tumors reached 5 mm in diameter,the mice were randomly di-

Expression of miR-29a in serum of patients with pulmonary tuberculosis and prediction of its function with bioinformatics analysis

Objective To analyze the expression of miR-29a in serum of patients with pulmonary tuberculosis,and to predict and analyze function of its target genes for further studying of its biological function and

Clinical features of chronic granulomatous disease: analysis of 27 cases

Objective To evaluate the clinical features and treatment of chronic granulomatous disease(CGD).Methods A total of 27 CGD patients diagnosed by neutrophil respiratory burst in Zhengzhou Children Hospital from January 2007 to June 2017 were selected.The clinical,pathogenic and radiological characteristics were retrospectively analyzed,and the treatment and prognosis were evaluated.Chi-square test was used for statistical analysis.Results Among the 27 cases,all patients presented with pulmonary infection(27/27)and 55.6% had cutaneous infection(15/27).Lymph node enlargement presented in 48.1%(13/27)of patients.Eleven neonates and four infants had cutaneous infection,which showed a significant difference(X^2=11.408,P<0.05).The results of pathogenic detection revealed 66.7%(18/27)fungal infection,25.9%(7/27)bacterial infection and 44.4%(12/27)mycobacterial infection.The incidence rate of fungal infection in neonates was significant higher than that of children(X^2=6.075,P<0.05).Radiologic examination showed that 12 cases had calcification of subaxillary lymph nodes,4 cases had pneumonia and thick-walled cavity,13 cases had multiple nodules,5 cases had massive high-density shadow and 4 cases had pulmonary abscess.The clinical pathogenic and radiologic features were similar between neonates and children.Treatment options included long-term combination therapy of bactericidal and fungicidal medicines.Conclusion Clinical manifestations of CGD are mainly pulmonary infection as well as cutaneous infection.Combination therapy with multiple medicines is effective.Hematopoietic stem cell transplatation plus retroviral vectors genetic therapy may be the trend of CGD treatment in the future.