Objective To evaluate modified Soave procedure for the treatment of Hirschsprung's disease for the neonate and infant. Methods Seventeen cases of short and sigmoid types of Hirschsprung's disease were treated by transanal modified Soave procedure. The incision was made 1cm posterioly and 2-3 cm anterioly above the dental line; the dissection progressed from the submucosa layer up to the perineal reflex into the pelvic cavity, and the rectum was pulled through. "V" shap muscular cuff was excised on the posterior wall. The proximal colon was anastomosed to the anal mucosa. Results Mean operative time was 160 min ? 45 min, mean blood loss was 45 ml ?35 ml. No enterocolitis, anastomotic leak, perianal infection and anastomotic stenosis occurred. All patients recovered with normal defecation, and no complication was observed from a follow-up of more than 4 mos. Conclusion Modified Soave procedure is safe and effective for the treatment of Hirschsprung's disease in the neonate and infant.

Objective To evaluate the influence of expression of pre S1 protein on the genomic expression of hepatitis B virus (HBV) infected hepatocyte with microarray. Methods The differentially expressed genes between the hepatoblastoma cell line HepG2 transfected by pcDNA3.1(-) and pcDNA3.1(-)-preS1, were respectively compared by cDNA microarray technique. The HBV pre-S1 coding DNA fragment was amplified with polymerase chain reaction (PCR) technique by using G376-7 plasmid DNA containing the full length of HBV genome as the template. The expressive vector of pcDNA3.1-preS1 was constructed by routine molecular biological methods. HepG2 cells were transfected by pcDNA3.1(-) and pcDNA3.1-preS1, respectively, using FuGENE6 Transfection Reagent. The total RNA was isolated and reversely transcribed. Results The cDNAs were subjected for microarray screening with 1152 cDNA probes. From the scanning results, it was found that 30 genes were up-regulated and 38 genes were down-regulated by pre-S1 protein of HBV. Conclusion The expression of pre-S1 protein affected the genomic expression spectrum of HBV infected hepatocyte(HepG2 cell line).

Objective To investigate the activity of HBV pre-S2 protein on thioredoxin reductase 1 (TXNRD1) gene promoter. Methods TXNRD1 gene promoter DNA sequence was identified in GenBank by bioinformatics and amplified from HepG2 genome by polymerase chain reaction (PCR). The amplified product was cloned into pCAT3-Basic reporter vector,named pCAT3-TXNRD1p. The HepG2 cells were transfected by pCAT3-TXNRD1p,and then co-tranfected by pCAT3-TXNRD1p and pcDNA3.1(-)-preS2 plasmids. The choloraphenical acetyltransferase(CAT)activity was assessed by enzyme linked immunosorbent assay(ELISA). Results The results indicate that HepG2 cells transfected by pCAT3-TXNRD1p had higher activity of CAT than that transfected by pCAT3-Basic. The expression of CAT in HepG2 cells co-transfected by pCAT3-TXNRD1p and pcDNA3.1(-)-preS2 was 2.2 times higher than that with pCAT3-TXNRD1p. Conclusions The TXNRD1 gene promoter identified in this study has transcription activity and HBV pre-S2 protein can transactivate the expression of TXNRD1 gene.

Objective To investigate the biological functions of HBeAg-binding protein 1(HBEBP1), suppression subtractive hybridization(SSH) technique was used to screen genes regulated by HBEBP1. Methods HBEBP1(GenBank number:AF529372) was screened and identified by yeast two-hybrid system 3 and co-immunoprecipitation technique. The HBEBP1 coding DNA fragment was amplified by reverse transcription polymerase chain reaction (RT-PCR) technique from HepG2 cell genome. The expressive vector of pcDNA3.1(-)-HBEBP1 was constructed by routine molecular biological methods. The HepG2 cells were transfected with pcDNA3.1(-) and pcDNA3.1(-)-HBEBP1, respectively by using FuGENE6 transfection reagent, then the mRNA was isolated. SSH method was employed to analyze the differentially expressed DNA sequences between the two groups. After restriction enzyme Rsa I digestion, small sized cDNAs were obtained. Then tester cDNAs were divided into two groups and ligated to the specific adaptor 1 and adaptor 2, respectively. After tester cDNAs were hybridized with driver cDNAs twice and underwent polymerase chain reaction (PCR) twice, they were then subcloned into pGEM-Teasy plasmid vectors to set up the subtractive library. Amplification of the library was carried out with E. coli strain DH5?. The cDNAs were sequenced and analyzed in GenBank with Blast search after PCR. Results The subtractive library of genes up-regulated by HBEBP1 was constructed successfully. The amplified library contained 85 positive clones. Colony PCR showed that these clones contained 200-1 000bp inserts. Sequence analysis was performed in 26 clones at random, and the full length sequences were obtained with bioinformatics method. Altogether 15 coding sequences were obtained. Conclusions The obtained sequences may be target genes up-regulated by HBEBP1, among which some genes coding proteins were involved in cell cycle regulation, metabolism, immunity and cell apoptosis. This finding brought some new clues for the study of the biological functions of HBEBP1 and HBeAg.

Computer science and technology has been used to promote the development of objectification of traditional Chinese medicine (TCM), which is also required for international development of TCM. In this paper, on the basis of current situation of Chinese medicine tongue objective research, the analysis was made on involved computer-related technology, relevant standards, and the future trend was discussed.

Objective To investigate the distribution and antimicrobial resistance of pathogens isolated from blood culture of children in a pediatric intensive care unit (PICU),provide reference for empirical treatment of bloodstream infection in critically ill children.Methods Pathogenic bacteria isolated from blood culture of children in a PICU in 2011-2015 were identified and performed antimicrobial susceptibility testing.Results A total of 180 strains of pathogens were isolated from 3 215 blood specimens,the positive rate was 5.60 ％,153 (85.00 ％) of which were grampositive bacteria and 27 (15.00 ％) were gram-negative bacteria.The top five isolated pathogens were Staphylococcus epidermidis (26.67 ％),Staphylococcus hominis (25.00 ％),Staphylococcus haemolyticus (11.66 ％),Escherichia coli (5.55 ％),and Staphylococcus aureus (3.89 ％).The resistance rates of Staphylococcus spp.to linezolid,vancomycin,and quinupristin/dalfopristin were all 0;the detection rates of methicillin-resistant coagulase-negative staphylococci (MRCNS) and methicillin-resistant Staphylococcus aureus(MRSA) were 70.18％ and 42.68％ respectively;Escherichia coli had high resistance rates to ampicillin,cefazolin,ceftriaxone,gentamycin,and compound sulfamethoxazole (50.00 ％-80.00 ％).Conclusion CNS and Escherichia coli are the main pathogens in blood culture of children in PICU,differences in antimicrobial resistance exist among different types of CNS.

Adult ; Aged ; Biomarkers ; blood ; Female ; Humans ; Laminin ; blood ; Male ; Middle Aged ; Occupational Exposure ; Risk Assessment ; Silicon Dioxide ; adverse effects ; Silicosis ; blood ; diagnosis ; Superoxide Dismutase ; blood ; Transforming Growth Factor beta ; blood

STUDY DESIGN: Changes of morphology and phenotype of cultured cells in media added lactate were observed. OBJECTIVES: To evaluate the effect of lactate on morphology and phenotype of cultured intervertebral disc cell. SUMMARY OF LITERATURE REVIEW: It was reported that lactate and pH were important factor in the degeneration of intervertebral disc. However the effect of lactate on morphology and phenotype of cultured intervertebral disc cell have not been studied. MATERIALS AND METHODS: Cells were dissociated enzymatically from rabbit nucleus pulposus. After attaining monolayer growth, the cells were incubated in media added 2mM or 5mM lactate. Total cell counts and morphological changes of the cells were periodically observed. Changes in cell phenotype were investigated by use of anti-collagen antibody stain. RESULTS: The cell groups added no lactate and 2mM lactate showed no difference in cell counts, morphology and phenotype. The cell group added 5mM lactate showed a reduction in final cell Counts and highel'ratio of fibroblastic cell in total population. Anti-collagen I Ab stained the Intra-and extra-cellular area of fibroblastic cells and intracellular area of chondrocytic cells. CONCLUSIONS: The current study suggests that high concentration of lactate inhibit intervertebral disc cell proliferation and accelerate morphological and phenotypical change to fibroblastic cell.
Cell Count ; Cell Proliferation ; Cells, Cultured ; Fibroblasts ; Hydrogen-Ion Concentration ; Intervertebral Disc* ; Lactic Acid* ; Phenotype*

Forty-two patients with type 2 diabetes mellitus (DM) were refered to 3 groups:type 2 DM without diabetic nephropathy (DN),type 2 DM with DN in the initial stage and type 2 DM with DN in the clinical stage.Ten healthy subjects were served as control group.Plasma adrenomedullin (ADM),urinary?_1- microglobulin (MG) and?_2-MG were detected.The results showed that the level of plasma ADM rose gradually with the development of DN and was positively correlated with markers of tubulointerstiial injury such as urinary?_1- MG and?_2-MG (both P

Objective Precondition is an approach to myocardial protection during ischemia-reper-fusion by inhibiting myocardial cell apoptosis.The purpose of this study was to evaluate the cardioprotective effect using 99Tcm-syuaptotagmin I (Syt I)-C2A to detect myocardial cell apoptosis in the myocardial is-chemia-repedusion rat model.Methods (1) The C2A domain of Syt I was labeled with 99Tcm using 2-iminothiophene hydrochloride (IT) method.Radiochemical purity was determined with thin layer chroma-tography.The binding activity of radiolabeled protein was assessed using eamptothecin-treated Jurket cells.(2) One group of 6 rats was prepared for myocardial ischemia-reperfusion model(A group),and another group of 6 rats was prepared for myocardial ischemia precondition model(B group).99Tcm-Syt I-C2A was injected via the tail vein at a dosage of about 7.4 MBq.At 1h after injection,the rat was sacrificed,and the heart was removed to rinse with saline and dye with triphenyl tetrazolium eoride (TTC).According to the resdt of myocardial dye,theischemic myoeardium was separated from the viable myocardium and weight was measured,and then its radioactivity was determined by gamma counting.The difference of radioactive uptake in the ischemic myocardium between these two group models was compared using percentage activity of injection dose per gram of tissue(%ID/g)±standard deviation(x±s).SPSS 12.0 was used for data analy-sis,and t-test was used to compare data.Results (1) The radiochemical purity of 99Tcm-Syt I-C2A was (98.90±0.43)%,and the radioactivity in the camptothecin-treated group was (10.99±0.55) folds higher than that of non-treated viable control group.(2)In the ischemia-reperfusion model,the radioactive uptake of 99Tcm-Syt,I-C2A was(2.41±0.32)%ID/g in the ischemic myocardium,and(0.16±O.02)%ID/g in the nomud myocardiunm.However,in the myocardial ischemia precondition model,(0.46±0.05)%ID/g in the isehemic myocardium was measured,and(0.20±0.05)%ID/g in the normal myocardium.Uptake of 99Tcm-Syt I-C2A in ischemic myocrdium showed statistically significant difference (t=8.52,P