Objective:To clone the outer membrane protein hopX gene of Helicobacter pylori(Hp)and to perform sequencing and analysis of biological information.Methods:Polymerase chain reaction(PCR)was used to amplify the hopX gene from Hp chromosomal DNA.Then the target gene was digested by restricted endonuclease enzyme of BamH I,and inserted into the prokaryotic expression vector pQE30 digested by corresponding restricted endonuclease enzyme.The recombinant vector was used to select and transform for nucleotide sequence analysis.The biological property at the amino acid level was analyzed by Omiga 2.0 and Antheprot v 5.0.The transformant colony was induced with IPTG and the fusion protein was analyzed by SDS-PAGE and Western blot.Results:The recombinant plasmid was constructed.DNA sequence analysis showed the sequence of hopX was 1 284 bp.The homology of the strains in nucleotide acid was 96%～97%.Their homogeneity in the amino acids was 97%～99%.We get a GeneBank accession number EF208122.Omiga 2.0 software predicted its relative molecular mass(Mr.)was 47 kD and possessed good antigencity.The expressed product contained about 37% of total somatic proteins and Western blot method showed good antigenicity of the recombinant protein.Conclusion:A confirmed gene hopX has been obtained,providing a good foundation for recombination,expression and related study.The corresponding peptide of the gene performed the structural characteristics of some typical antigen molecules,which suggest that it might be a novel vaccine candidate.

Objective To explore the concentration of mannan-binding lectin(MBL ) of patients with chronic hepatitis B virus (HBV) .Methods Serum MBL concentrations of 250 patients(case group) with HBV and 150 healthy controls(control group) were measured .Results The serum MBL concentration in case group was higher than that in control group(t=7 .097 ,P<0 .01) . The serum MBL concentration in high HBV-DNA loading group was higher than that in control group(t=7 .179 ,P<0 .01) .The serum MBL concentration in low HBV-DNA loading group was higher than that in control group(t=4 .404 ,P<0 .01) .Conclusion Detection of serum MBL in patients with HBV will be clinically useful for understanding state of an illness and observing the cur-ative effect .

Objective To study the expressions of IL-1 receptor type Ⅰ(IL-1RI) mRNA and IL-1? protein in the rat carotid body. Methods In situ hybridization,immunofluorescence double staining and Western blotting methods were used. Results The result of in situ hybridization showed that the positive signal of IL-1? mRNA was mainly located in the glomus cells of the carotid body.The result of immunofluorescence double staining showed that IL-1? protein also expressed in the glomus cells of the organ.The Western blotting proved that the IL-1? immunoreactive band appeared at 18kD,consistent with the molecular weight of the cytokine.Conclusion The glomus cells of the rat carotid body not only express IL-1RI mRNA,but also IL-1?.

AIM: To study the effects of 1.5 MAC halothane and sevoflurane on ischemic myocardium. METHODS: The isolated rat heart were perfused with halothane and sevoflurane and HR, LVEDP, LVDP, +d p /d t , -d p /d t , coronary flow (CF), the myocardial ATP content and Ca 2+ -ATPase activity were determined before and 10 min and 25 min after ischemia. In the meantime, LVP was recorded during 25 min ischemia. RESULTS: 1.5MAC sevoflurane significantly increased CF in normal isolated rat hearts. Both halothane and sevoflurane depressed myocardial contractile function, increased normal myocardial energy storage. After 10 min ischemia, the decrease of myocardial ATP content were slowed down by halothane and sevoflurane, especially halothane. During 25 min of ischemia, the onset time of contracture was significantly delayed, and the contracture intensity was alleviated by halothane, but not sevoflurane. CONCLUSION: Halothane has better protective effect on ischemic myocardium than sevoflurane through preventing the decrease of myocardial ATP content and Ca 2+ -ATPase activity during ischemia.

BACKGROUND:Among many transplanted cells,adult autologous bone barrow-derived mononuclear cells have beenused in clinical practice because they are easy to be obtained,without immunological rejection and ethical disputationand other advantages.How to distinguish donor cells from receptors and observe the survival of donor cells following stem cell transplantation still trouble people.OBJECTIVE: superparamagnetic iron oxide (SPIO)particles-labeled bone marrow-derived mononuclear cells from minipigs were used to observe the feasibility of in vivo tracking with magnetic resonance imaging(MRI).DESIGN:A controlled observation experiment.SETTING:Institute of Cardiovascular Disease,Zhongda Hospital Affiliated to Southeast University.MATERIALS:This experiment was carried out in the Institute of Cardiovascular Disease,Zhongda Hospital Affiliated to Southeast University between April 2006 and August 2006.Healthy Chinese minipigs,aged 3 to 4 months,weighing from 20 to 30 kg,were provided by the Experimental Animal Center of Southeast University[SYXK(Su)2002-0012].METHODS: Autologous bone marrow-derived mononuclear cells of minipigs were isolated and cultured. Bone marrow-derived mononuclear cells in the suspension were traced with SPIO particles.Ferrum in the cells were shown by Prussian blue staining, and cell viability was evaluated by trypan blue exclusion method. Eleven minipigs used for preparation of model of myocardial infarction were divided into experimental group(n=9)and control group(n=2).By means of percutaneous left or right cervical artery or femoral artery puncturation, 1.5 to 2.0 mm balloon was used to occlude 1/3 left anterior descending branch,304 to 405 kPa,60 minutes later,ischemic preconditioning was conducted 3 tO 4 times before operation. When pig models of myocardial infarction were successful that was proved by surface electrocardiogram,bone marrow-derived mononuclear cells were percutaneously injected into coronary artery.Coronary arteriography was performed through femoral artery acupuncture at 24 hours after establishing infarction models.Suspension of bone marrow-derived mononuclear cells was perfused into coronary artery with OTW catheter.Then,the injector and OTW catheter for containing cells were rinsed with normal saline containing heparin and infused with the residual cells within 10 minutes.Non-labeled cells were perfused in 2 minipigs of control group by the same method.Postoperatively, bone marrow-derived mononuclear cells were traced by magnetic resonance and compared with Prussian blue-stained myocardial tissue sections.RESULTS: Seven minipigs of experimental group and one minipig of control group were Involved in the final analysis.One of each group was used for preparation of model of myocardial Infarction.One minipig of experimental group died from anesthetic accident before magnetic resonance.①Bone marrow-derived mononuclear cells all were nearly labeled by SPIO particles. Bone marrow-derived mononuclear cells could further proliferate in culture medium containing Fe2O3-PLL without obvious changes of cellular shape. ②T2+WI showed that 5 of 8 models of myocardial infarction presented fuzzy low-echo signal region in peripheral myocardial infarction after transplantation of labeled cells and the low-echo signal disappeared 4 weeks Iater. Ex vivo T2+WI sequence showed there was a dot-distributed low-echo signal region in the peripheral infarction region.③It was found in histological examination that 5 models(cell number over 106) had Prussian blue-positive cells,which distributed the same as those in magnetic resonance signal reducing region.CONCLUSION:SPIO particles-labeled bone marrow-derived mononuclear cells are safe and effective;T2+ WI is sensitive to tracing SPIO particles-labeled bone marrow-derived mononuclear cells;Magnetic resonance can in vivo trace SPIO particles-labeled stem cells transplanted through coronary artery,magnetic resonance signal change is related with the number of stem cells and division growth.

Heat shock protein gp96 isolated from tumor tissues holds great promise for tumor immunotherapy. However, at present only very limited amount of gp96 protein can be isolated from tumor tissues. Here, we reconstituted the yeast-expressed gp96 (recombinant gp96, rgp96) with B16.F10 melanoma antigens in vitro to prepare new gp96 tumor vaccine on large-scale, and analyzed its induction of specific anti-tumor immunoresponses by ELISPOT, IFN-gamma intracellular staining and cytotoxicity assays. Immunization with rgp96-tumor antigen complexes significantly inhibited B16 tumor growth compared with either rgp96 or tumor antigens alone and led to enhancement of tumor-specific T-cell activities, which was found similar to that of tumor tissue derived gp96. Our results therefore may provide bases for large-scale preparation of the new generation of gp96 tumor vaccines.
Animals ; Cancer Vaccines ; biosynthesis ; genetics ; immunology ; therapeutic use ; Female ; Heat-Shock Proteins ; biosynthesis ; genetics ; immunology ; therapeutic use ; Melanoma, Experimental ; therapy ; Mice ; Mice, Inbred C57BL ; Neoplasm Transplantation ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; therapeutic use ; Skin Neoplasms ; therapy ; Yeasts ; genetics ; metabolism

Objective:To investigate the expression and its diagnostic value of long-chain non coding RNA (lncRNA) and colon cancer-related transcript-2 (CCAT2) in serum of patients with cervical cancer (CC).Methods:Serum samples of 100 CC patients, 60 CIN patients and 80 healthy people enrolled by Nantong Tumor Hospital from January 2016 to June 2017 were collected.The expression levels of CCAT2 in sera of CC patients and their corresponding postoperative patients, CIN patients and healthy controls were detected by real-time fluorescent quantitative PCR (RT-qPCR). The correlation between CCAT2 and clinicopathological features, as well as the traditional auxiliary diagnostic makers of CC, such as carbohydrate antigen 125 (CA125) and squamous cell carcinoma antigen (SCC) was analyzed. The working characteristic curve (ROC) of the subjects was used to evaluate the CCAT2 pair in the auxiliary diagnostic value of cervical cancer.Results:The relative expression levels of serum CCAT2 in patients with cervical cancer, patients after operation, patients with CIN and healthy controls were1.689 (0.616, 4.776), 1.018 (0.227, 3.328), 0.815 (0.453, 1.266) and 0.740 (0.271, 1.670), respectively.The relative expression of CCAT2 in cervical cancer patients was significantly higher than that in post-operative patients, CIN patients and healthy controls. The difference was statistically significant( t=6.999,8.193,9.345, P<0.001).There was no significant difference in the relative expression of CCAT2 between CIN patients and healthy controls ( t=0.327, P>0.05).The relative expression of CCAT2 in serum of cervical cancer patients had no significant difference in age 1.636(1.000,2.370),1.705(1.095,2.243) (χ 2=0.137, P=0.712) and menopause 1.672(1.059,2.342),1.659(1.068,2.298) (χ 2=0.000, P=1.000), but had significant difference with tumor size expression1.189(0.916,1.725),2.019(1.537,2.497)(χ 2=17.508, P=0.000),International Federation of Obstetrics and Gynecology (FIGO) staged expression stage 0.993(0.779,1.266),2.056(1.547,2.549),3.987(3.699,4.275)(χ 2=36.075, P=0.000) and lymph node metastasis 1.434(1.007,2.251),2.019(1.731,3.098) (χ 2=8.634, P=0.003). There was no correlation between the relative expression of serum CCAT2 and CA125 ( r2=0.003, P=0.563) and SCC (r 2=0.128, P=0.000).The diagnostic efficacy of serum CCAT2, CA125 and SCC in cervical cancer patients was analyzed by ROC curve. When compared with CIN patients, the areas under the curve were 0.890, 0.549 and 0.744, respectively. When compared with healthy patients, the areas under the curve were 0.857, 0.650 and 0.758, respectively. Conclusions:The level of serum CCAT2 in patients with cervical cancer is significantly higher than that in patients with cervical cancer, CIN patients and healthy controls. Serum CCAT2 may be a relevant marker for the diagnosis and prognosis of cervical cancer.

Objective To explore the effect of quality control circle ( QCC ) on non-standard implementation of aseptic operation related medical staffs. Methods Through QCC activities, a total of 950 operation related medical staffs had been investigated non-standard implementation status of aseptic operation, and analyzed the causes, made out and carried out its countermeasures. Results Non-standard implementation of aseptic operation declined from 690 person-time to 175 person-time, and the target completion rate was 105%, improvement rate 75%, of which spreading towels did not reached standard declined from 327 person-time to 99 person-time. The off operation table people, who was too near from the sterile stage, declined from 134 person-time to 33 person-time, while not using the non-contact wear gloves declined from 71 person-time to 12 person-time. Conclusions QCC activities can significantly reduce the person-time of non-standard implementation of aseptic operation and effectively improve the quality of work.

OBJECTIVETo evaluate the risk factors associated with pathogenesis of cerebral palsy (CP) in young children.

METHODSA cross-sectional survey on the prevalence of CP was conducted in young children aged one to six years in Leshan area, Sichuan Province. Cluster sampling and a 1:2 case-control design were used to investigate the risk factors for pathogenesis of CP.

RESULTSTotally, 148,723 children were surveyed, in which 308 (2.07 per thousand ) were diagnosed as CP. Low birth weight, twins and premature birth were associated with significantly increased prevalence of CP, 16.32, 4.16 and 22.21 times as that in normal birth weight, single birth and full-term birth, respectively. Simple factor analysis showed varied factors involved in pathogenesis of CP. The multivariate analysis revealed that delivery at home, low Apgar score in five minutes, illness during the first month of life, maternal "cold" with fever in their early gestation, low protein (meat and egg) intake during pregnancy and lower education level of mother were risk factors for CP.

CONCLUSIONSPrevalence and clinical features of CP in Leshan was comparable to those in the developed countries. Relevant risk factors could be seen primarily in gestational and perinatal periods, which might involve in mothers, children, environment and heredity, etc. To attach more importance in gestational and perinatal care for mothers and babies will be crucial measures to reduce occurrence of CP in young children.

Case-Control Studies ; Cerebral Palsy ; epidemiology ; Child ; Child, Preschool ; China ; epidemiology ; Humans ; Infant ; Infant, Low Birth Weight ; Infant, Newborn ; Infant, Premature ; Multivariate Analysis ; Prevalence ; Risk Factors

Objective:To describe the prevalence of food allergy among children aged 0-5 years in China and to explore related influencing factors.Methods:Multistage stratified random sampling method was used to collect data from 275 surveillance sites of the China National Nutrition and Health Survey of Chinese children and lactating mothers programs in 31 provinces (autonomous regions and municipalities) of China in 2016-2017. A total of 70 107 participants aged 0-5 years were included in this study. The study collected information of participants' demographic characteristics and food allergies by face-to-face questionnaire. The prevalence of food allergy was analyzed, using the complex data weighting method. The logistic regression models were used to analyze the influencing factors related to food allergy.Results:The overall prevalence of self-reported food allergy among children aged 0-5 years was 4.81%. Prevalence rates in infants aged 0-5 months, and 6-23 months and preschool children aged 2-5 years were 0.81%, 4.68% and 5.26%, respectively. The results of logistic analysis showed that there was a significantly positive correlation between factors including children from 6 months to 5 years old, urban area, southwest area, first-born, mothers with college education or above, and the prevalence of food allergy in children. Shrimp, poultry eggs, crab shellfish, fruit, milk and fish appeared the common allergic foods in children aged 0-5 years, with prevalence rates of self-reported food allergy as 1.55%, 1.25%, 0.99%, 0.97%, 0.87% and 0.86%, respectively. The proportion of single food allergy in children with allergies was 69.85%.Conclusions:Among children aged 0-5 years, the prevalence of self-reported food allergy increases with age, in China. Foods that is prone to allergies include fish, shrimp, crab, shellfish, poultry eggs, milk and fruits, etc. Most allergies were only caused by single food in children, under observation.