Objective To clone and express translocation intimin receptor(Tir)of enterohemorrhagic Escherichia coli(EHEC)O157:H7,and to analyze the effect of different routes on the induction of immunity to the recombinant protein.Methods The tir eucoding genes were amplified from EHEC O157:H7 strain guangzhou 246 genome,and genes were cloned into the vector pET-30a(+).The pET-30a(+)tir recombinant was transformed into E.coli BL21.and expression was induced bv IPTG.The expressed product was analyzed by SDS-PAGE and purified by Ni-IDA affinity chromatography.The immunized mice sera and fecal against the recombinant protein was detected.Resuits The length of the tir is 1677 bp,with the initiation codon ATG and the termination codon TAA.Double enzyme digestion and DNA sequencing confirmed that the recombinant expression plasmid pET-30a(+)-tir was constructed.The recombinant protein was expressed in Escherichia coli expression system,and was purified by Ni-IDA affinity chromatography.The mice were able to produce a high serum IgG antibody titer after both subeutaneous and intranasal immunizations.Meanwhile,the intranasal immunization induced serum and fecal IgA antibody titer was significantly higher than that of the subcutaneous immunization group.Conclusion Tir molecule is potential vaccine candidate for preventing EHEC disease.

Objective To evaluate uterine cavity of the patients who accept the treatment in vitro fertilization‐embryo trans‐fer(IVF‐ET)and determine the clinical value of hysteroscopy for the patients .Methods Between January 2012 and May 2014 ,539 cases who accepted the IVF‐ET and hysteroscopy ,were divided into two grops ,group A(258 patients examinated before IVF‐ET cycle) and group B(281 patients after the IVF‐ET failure) ,and retrospective analysis was performed for their uterine cavity and cervix canal data .Results The total incidence of abnormal hysteroscopic findings was 49 .54% ,and group B was significantly high‐er than that in group A(54 .10% vs .44 .57% ,χ2 =4 .5 ;P=0 .043) .The percent of abnormal cervix canal was 18 .74% ,and there wasn′t significant difference between the two groups .Conclusion The incidence of abnormal hysteroscopic findings in IVF‐ET is higher than that in normal patients ,so hysteroscopy has an important clinical value for diagnosis and evalution ,especially for pa‐tients with failure of IVF‐ET .

Objective To explore anti-EV71 effects of Ganlu Xiaodudan in vitro. Methods Ribavirin was taken as control drug, with the help of cell culture to observe anti-EV71 inhibition rate of Ganlu Xiaodudan in inhibiting-virus-directly experiment, therapeutic-inhibiting-virus experiment, preventive-inhibiting-virus experiment and preventive-therapeutic-inhibiting-virus experiment. Results In inhibiting-virus-directly experiment, therapeutic-inhibiting-virus experiment and preventive-therapeutic-inhibiting-virus experiment, virus inhibition rate of Ganlu Xiaodudan was higher than ribavirin. In preventive-inhibiting-virus experiment, virus inhibition rates of Ganlu Xiaodudan and ribavirin both were almost zero. Conclusion Ganlu Xiaodudan has better antiviral effects on EV71 than ribavirin, and it can affect more than one link of multiplication of EV71.

Signal transducer and activator of transcription (STAT) 3 is a critical transcription factor for cell proliferation and survival. It is activated within cells by many cytokines to mediate immune and inflammatory responses to injury. Inflammatory bowel disease (IBD), represented by Crohn′s disease (CD) and ulcerative colitis (UC), is a chronic inflammatory disease of the intestinal tract. STAT3 has been shown to be abnormally activated in IBD colon tissues by many pro-inflammatory cytokines, leading to disruption of the intestinal mucosal barrier and excessive innate immune and Th17 responses. The persistent chronic inflammation eventually leads to intestinal fibrosis and stenosis. In addition to immune responses, STAT3 is also involved in intestinal fibrosis in IBD by promoting the transcription of fibrosis-related genes. Colitis-associated cancer (CAC) is a particularly aggressive subtype of colorectal cancer and is associated with chronic inflammation-induced IBD. STAT3 has also been associated with CAC initiation and development. STAT3 is overactivated in tumors, which leads to suppression of the anti-tumor activity of immune cells and promotion of cancer cell proliferation, tumor angiogenesis, invasion, and migration. In the present article, we summarize the role of STAT3 in IBD and CAC and the research progress of the related drugs developed for UC and CAC treatment.

Acute Kidney Injury ; therapy ; Cardiac Surgical Procedures ; adverse effects ; Female ; Humans ; Infant, Newborn ; Male ; Peritoneal Dialysis

OBJECTIVETo investigate the evolution of hepatitis B virus (HBV) quasispecies in one patient during lamivudine (LAM) monotherapy and switching to entecavir (ETV) rescue treatment.

METHODSSerum samples were taken at seven different time points during antiviral therapy (0, 24, 48, 60, 72, 96, 152 weeks, respectively), the HBV DNA polymerase gene was amplified, cloned and sequenced to analyze the amino acid substitutions within HBV DNA polymerase gene and distribution of virus quasispecies. Quantitative detection of the HBV wild strains and total virus was performed by amplification refractory mutation system real-time PCR (ARMS-PCR).

RESULTSThree mutation patterns detected during antiviral therapy in the patient: rtM204V, rtM204V+rtL180M and rtM204I. The HBV quasispecies were found always in dynamic variation. The HBV populations were completely replaced with the LAM-resistant variants when the viral breakthrough was encountered during LAM monotherapy. Interestingly, the wild-type variants presented gradually dominant (79.3%) with the decline of HBV DNA load after switching to ETV rescue administration. ARMS-PCR results showed that the wild-type variants account ed for 68.55% of the HBV populations at baseline and this proportion declined to 0.21% when the viral breakthrough emerged under LAM therapy. The wild-type variants gradually increased from week 24 after switching to ETV rescue therapy and the proportion of HBV wild-type variants in the population fluctuated between 16.01% to 26.93%.

CONCLUSIONSThe distribution of virus quasispecies were always in dynamic variation during sequential therapy with nucleotide analogs in chronic hepatitis B patients. Different patterns of dynamic HBV quasispecies may have different contribution in ETV resistance in LMV refractory patients with ETV administration.

Adult ; Antiviral Agents ; therapeutic use ; DNA, Viral ; genetics ; Drug Resistance, Viral ; genetics ; Genotype ; Hepatitis B virus ; drug effects ; genetics ; Hepatitis B, Chronic ; drug therapy ; virology ; Humans ; Male ; Mutation

OBJECTIVETo evaluate the relationship between mitochondrial DNA instability (mtMSI) and interleukin-8 (IL-8) activity in gastric mucosa of various lesions.

METHODSIL-8 level in gastric mucosa was assayed using ELISA method. The mtMSI was detected by PCR-SSCP techniques.

RESULTSmtMSI was observed in 11 out of 30 (36.7%) gastric cancers, 2 of 15 (13.3%) intestinal metaplasia, 2 of 10 dysplasia and 1 of 10 chronic atrophic gastritis. IL-8 level in mtMSI+ group [(76.8 +/- 3.8) pg/mg] was significantly higher than that in mtMSI- group [(48.3 +/- 3.6) pg/mg, P < 0.05].

CONCLUSIONmtMSI closely correlates with IL-8 level in gastric mucosa and is involved in gastric carcinogenesis.

DNA, Mitochondrial ; genetics ; Enzyme-Linked Immunosorbent Assay ; Gastric Mucosa ; metabolism ; pathology ; Gastritis, Atrophic ; genetics ; metabolism ; Genomic Instability ; Humans ; Interleukin-8 ; metabolism ; Metaplasia ; genetics ; metabolism ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; Precancerous Conditions ; genetics ; metabolism ; Stomach Neoplasms ; genetics ; metabolism

OBJECTIVETo quantitatively evaluate the murine model of spermatogenesis regeneration induced by two-dose busulfan injection.

METHODSFifty-four male mice were randomly divided into a control and two model groups of equal number, the former treated by two-dose intraperitoneal injection of 50% DMSO solution at 10 ml/kg, and the latter by that of busulfan at 10 mg/kg and 15 mg/kg respectively to establish spermatogenesis regeneration models, both at the interval of 24 days between the two doses. Spermatogenesis in seminiferous epithelia was evaluated by Johnsen score, and the expressions of GATA-4 and GDNF mRNA in Sertoli cells were detected by real time quantitative PCR at 3, 4 and 8 weeks after the treatment.

RESULTSJohnsen score kept stable in the control group at all stages (P > 0.05), but higher than in the model groups at 3 and 4 weeks (P < 0.01). It was lower in the 15 mg/kg than in the 10 mg/kg model group at 4 and 8 weeks (P < 0.01) , and than in the control group at 8 weeks (P < 0.05), but had no significant difference between the 10 mg/kg and the control groups (P > 0.05). Nor did the expression of GATA-4 mRNA in Sertoli cells show any significant difference among the three groups at different stages after the treatment (P > 0.05), and that of GDNF mRNA at different stages in the control group (P > 0.05). Compared with the controls, the level of GDNF mRNA in Sertoli cells was significantly higher at 3 weeks but lower at 4 weeks in the model groups (P < 0.01), and lower in the 15 mg/kg group (P < 0.01) and comparable in the 10 mg/kg group at 8 weeks (P > 0.05); and it was lower in the 15 mg/kg than in the 10 mg/kg group at all stages (P < 0.01).

CONCLUSIONTwo-dose intraperitoneal injection of 10 mg/kg busulfan at the interval of 24 days is an optimal option for the establishment of a murine model of spermatogenesis regeneration. Higher dose of busulfan may induce deficient expression of GDNF in Sertoli cells and result in incomplete restoration of spermatogenesis.

Animals ; Busulfan ; adverse effects ; Glial Cell Line-Derived Neurotrophic Factor ; metabolism ; Male ; Mice ; Mice, Inbred Strains ; Models, Animal ; RNA, Messenger ; Regeneration ; drug effects ; Sertoli Cells ; drug effects ; Spermatogenesis ; drug effects ; Spermatozoa ; physiology ; Testis ; drug effects ; physiology

Objective To investigate the glucose tolerance and ability of insulin secretion in SD rats in second and third trimesters of pregnancy. Methods SD rats with pregnancy of 15 d were selected as experimental group(n=6),and another 6 rats of the same batch without pregnancy were served as controls(n=6).Intraperitoneal glucose tolerance trials(IPGTT) were conducted in these two groups.Rat islets were isolated after in situ collagenase digestion through pancreatic duct perfusion,islet morphology was observed by inverted phase contrast microscope,and insulin secretion was determined by radioimmunoassay. Results It was revealed by IPGTT that the levels of glucose at 30,60,90 and 120 min were significantly lower in experimental group than those in control group(P

Human parathyroid hormone peptide1-34(hPTH1-34) was highly expressed in Escherichia coli by inserting the synthesized hPTH1-34 cDNA into pThioHis, the prokaryotic expression vector. The expressed hPTH1-34 was purified by chelating sepharose immobilized metal ion affinity, reverse and filter chromatographic steps. Its purity was verified above 95% by HPLC. The quality was identified by N-terminal sequencing and MALDI-TOF-MS analysis. In vitro analysis showed the adenylate cyclase of ROS 17/2.8 cells was activated by hPTH1-34.