Objective To establish a polymerase chain reaction method for the detection of the rcp virulence gene of L. pneumophila. Methods Sixteen strains of Legionella pneumophila, L. dumiffii, L. bozemanii and L. Longbeachae isolated from the cooling towers of the centralized air-conditioning systems in Tianjin form 2005 to 2007 were detected by polymerase chain reaction method using L. pneumophila rcp virulence gene-specific primer according to GenBank published nucleotide sequence. Results The 900 bp rcp genetic fragment was amplified in three strains of L. pneumophila, while others not. Conclusion The rcp virulence gene-based polymerase chain reaction method for the detection of L. pneumophila has been successfully established and is the foundation for the studies of Legionella virulence.

Objective To construct the multi-epitope antigen(mea) gene of G2 glycoprotein of Hantavirus SEO type L99 strain.Methods The B cell epitopes was chosen and connected by three-peptide GPG after the amino acid sequence of G2 protein was analyzed and predicted by bioinformatical soft wares.The corresponding gene mea was constructed by overlap PCR,and cloned into the prokaryotic expression plasmid pET32a(+).Results The mea gene was successfully constructed by the five epitopes being chosen.The recombinant plasmid pET32a-mea was acquired by directional cloning.Conclusion The mea and its expression system E.coli BL21/pET32a-mea were constructed for the first time.The foundation was laid for the expression of the mea and its immunological applications.

Objective To report a new minimally invasive and cosmetic approach for atrial septal defect repair. Methods 46 patients ranged 3.6 to 32 (12.5?7 7) years underwent minimal right vertical infra axillary thoracotomy for atrial septal defect repairs under beating hearts from January 1997 to March 2000. One had a functional single atrum,two had an associated partial anomalous pulmonary venous connection,and three had the moderate pulmonary hypertension. Results The average bypass time was (30 3?7 8) min. There was no operative or late mortality and no morbidity directly related to the thoracotomy with beating heart. 37 patients had been Followed up for 3 months to 2 4 years(1 3?0 6) years. All of patients were free of symptoms. Conclusions The minimal right vertical infra axillary thoracotomy is a safe,cosmetic and minimal invasive approach to median sternotomy for repair of atrial septal defect or anomalous pulmonary venous connection.

Objective To evaluate the role of autophagy on acute kidney injury after liver transplantation.Method Fifty-six healthy male Sprague-Dawley rats were randomly assigned into 4 groups:sham group,orthotopic liver transplantation (OLT) group,sirolimus pretreated (SRL) group and 3-methyladenine pretreated(3-MA) group.OLT model was established.Then the rats were sacrificed at 6 h after reperfusion.The renal function and the extent of oxidative stress relative proteins malondialdehyde (MDA) and superoxide dismutase (SOD) were observed.The levels of apoptosis relative genes caspase-3 and cyt c and the expression of autophagy relative proteins were detected.The pathological changes were microscopically examined in renal tissues.TUNEL staining was used to observe the apoptosis of tubular epithelial cells.Transmission electron microscopy was applied to observe the ultrastructure changes of tubular epithelial cells.Result As compared with sham group,OLT and 3-MA groups showed a serious renal injury including cellular vacuolization,loss of brush borders,and a significant rise in BUN,Cr and MDA,while a decrease in SOD activity.The levels of caspase-3 mRNA and cyt c rnRNA were increased significantly.Whereas compared to OLT and 3-MA groups,renal function and oxidative stress levels in SRL group ameliorated,and histopathologic damage and apoptosis alleviated after OLT.Simultaneously,the levels of caspase-3 mRNA and cyt c mRNA were decreased.The expression of beclin-1 and LC3-]Ⅱ was effectively upregulated.Conclusion Autophagy could alleviate acute kidney injury after liver transplantation through inhibiting oxidative stress and apoptosis.

Objective To study proteome variation between uropathogenic E. coil (UPEC)132, UPEC J96 and non-uropathogenic E. coli K-12 MG1655. Methods Two-dimensional gel electrophoresis(2-DE) was applied to compare the differential expression proteins between UPEC 132, UPEC J96 and non-uro-pathogenic E.coli K-12 MG1655. The differential expression proteins were digested in gel by enzyme. The mass of generated peptides were measured by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). The data obtained from peptide mass fingerprinting (PMF) were re-searched using the internet available database. Results The number of protein spots recognized from UPEC 132 was 466±11, significantly more than that of E. coli K-12 MG1655 (338±15) and UPEC J96 (382±12); there were 298 protein spots shared by the three E.coli strains, 56 protein spots shared by two UPEC strains, and 89 protein spots characterized by UPEC 132. Twenty-two differential expression or significantly increased expression protein spots, involved in virulence factors, metabolism and transportation, regulation of protein synthesis, biological oxidation and unknown functions, were successfully identified by MALDI-TOF-MS. Condusion The proteome from UPEC 132 and non-uropathogenic E. coli K-12 MG1655, or UPEC 132 and UPEC J96 was differentially expressed. It will provide important information on the pathogen-esis of UPEC 132.

A method of streptomycin resistance screening was applied to improve t he productivity of Natamycin by Streptomyces gilvosporeus(ATCC13326) The sp ores treated with UV light were regenerated on agar plates containing 0 6?g/mL stre p tomycin 122 streptomycin resistant(str) mutants were obtained The Natamycin y iel ds of 13 mutants were higher than the original strain The mutants with high Na t amycin productivity were screened at a high frequency(10 6%) The highest one that demonstrated 1 46 times that of the original strain in Natamy cin productivity was obtained

Objective To establish RT-PCR-RFLP method for studying the genotype of wild mea-sles virus strains isolated from Tianjin area from 2002 to 2008. Methods Isolations of measles virus were carried out by tissue culture method from urine and throat swab specimens collected from suspected cases. RNA were extracted from the virus specimens. The 594 bp fragment of C terminal of the N (nucleoprotein) gene was amplified by one-step RT-PCR, then the PCR products were digested with Bcn I , separated on agarose gel electrophoresis and then analyzed by the method of RFLP (restriction fragment length polymor-phism). In addition, above results were compared with DNA sequencing. Phylogenetic tree was plotted based on the results for the genetic relationship and distance analysis. Results Sixty-nine measles virus strains were isolated from 189 specimens from 2002 to 2008, of which the C terminals of N gene were all de-tected positive. Among the 69 strains of measles virus isolates, 98.55% (68/69) belonged to Hla sub-geno-type which was the predominant sub-genotype, and only one strain (1.45%) belonged to H1b sub-genotype by RFLP analysis which was in accordance with the results by DNA sequencing method. Phylogenetic tree analysis indicated the H1a sub-genotype measles virus strains should be further divided into 2 clades, and the variation fluctuated between 0.2% and 3.8%. There were transmission chains caused by different virus strains co-cireulation. Conclusion A genotype, H1a and H1b sub-genotype can be identified by RT-PCR-RFLP assay specically based on the restriction enzyme Bcn I .The RT-PCR-RFLP assay can be a rapid, simple, accurate and efficient method for large-scale surveillance of measles virus strains in China.

Objective:To research the clinical effects of ulinastatin in the treatment of sepsis induced acute renal injury and its possible mechanisms.Methods:114 cases of patients with sepsis induced acute kidney injury from 2014.02 ～ 2016.08 were selected and randomly divided into the control group (n=57) and experimental group (n=57) according to the draw method,the control group was given conventional treatment,while the experimental group was treated by ulinastatin based on the control group,the urine urinary injury molecule-1 (KIM-1),atrialnatriuretic peptide (ANP),cyscatin-c (CYS-C),interleukin l,6 (IL-1,IL-6),c-reactive protein (CRP),tumor necrosis factor-α(TNF-α),nitric oxide (NO),endothelin 1 (ET-1),immunoglobulin A,G,M (IgA,IgG,IgM) levels,APACHE-Ⅱ score were compared between two groups before and after the treatment.Results:After treatmented,the urine of KIM-1,ANP,serum of CYS-C,IL-l,IL-6,CRP,TNF-α,ET-1 levels and APACHE-Ⅱ score of experimental group were significantly lower than those of the control group (P＜0.05).The serum NO,IgA,IgG,IgM levels of experimental group were significantly higher than those of the control group (P＜0.05).Conclusion:Ulinastatin could significantly relieve sepsis induced acute renal injury,which might be related to the inhibition of inflammatory response,improvement of the renal blood flow and immune function.

Objective To explore the intervention effect of ketogenic diet (KD) on neurobehavioral damages after flurothyl-induced neonatal recurrent seizures in rats and on the expression of ZIP7.Methods Postnatal day 8 SD rats (n=24) were divided into four groups randomly:normal group (NS+ND group),non-seizure and ketogenic diet group (NS+KD group),seizure and normal diet group (RS+ND group),seizure and ketogenic diet group (RS+KD group),n=6 in eacb group.At postnatal day 31,the grip-strength test and open field test were monitored.At postnatal day 32,rats were sacrificed and the expression of ZIP7 protein level in cerebral cortex was detected with Western blot.Results (1) The grip-strength test:compared with NS+ND group ((32.67±2.42) s),the time needed to hold on glass bar in RS+ND group ((19.17±2.48) s) was shorter significantly (P＜0.05).Compared with RS+ND group,the time needed to hold on glass bar in RS+KD group ((26.25±2.87) s) was significantly longer (P＜0.05).(2) Open field test:compared with NS+ ND group ((2.00± 0.63) times),the times of grooming in RS+ND group ((4.00±0.63) times) were more (P＜0.05).Compared with RS+ND group,the times of grooming in the RS+KD group ((2.17±0.75) times) were fewer (P＜0.05).(3)Western blot:compared with NS +ND group,the level of ZIP7 of the RS+ND group in cerebral cortex were lower (P＜0.05).Compared with RS+ND group,the level of ZIP7 of the RS+KD group in cerebral cortex were higher (P＜0.05).Conclusion Neonatal recurrent seizures may damage neurobehavior,and the neuroprotective effects of ketogenic diet may be associated with the increasing of ZIP7 in cerebral cortex.

Objective To analyze the characteristics of antibiotic resistance in group A Streptococ-cus ( GAS) strains isolated from children with scarlet fever in Tianjin in order to provide reference for clinical drug administration. -ethods GAS strains were collected from 2011 to 2016. A total of 276 isolates were analyzed by antibiotic susceptibility test and emm typing. Results All of the isolates were susceptible to penicillin, cefazolin and vancomycin, while 98. 2％ were susceptible to both chloramphenicol and levofloxa-cin. The resistance rates to azithromycin, erythromycin, clarithromycin, clindamycin and tetracycline were 97. 8％, 97. 1％, 94. 2％, 94. 2％ and 79. 3％. The concomitant resistance to erythromycin, azithromycin, clarithromycin, clindamycin and tetracycline was 73. 2％. The resistance rates of GAS strains isolated from different years to tetracycline, clindamycin, clarithromycin, erythromycin and azithromycin were significantly different. A statistically significant difference was found between the percentages of emm12 and emm1 strains resistant to tetracycline (84. 0％ vs 59. 5％, χ2=13. 820, P=0. 000). Conclusions The isolated GAS strains are sensitive toβ-lactams and highly resistant to macrolide antibiotics, clindamycin and tetracycline. Penicillin remains the preferred treatment for GAS infection and cephalosporins may be used as a substitute if the patient is allergic to penicillin.