Objective To construct a qseC-deleted mutant strain of E.coli by Red recombination and to study the effect of qseC gene on biofilm formation in the mutants.Methods The chloramphenicolresistant gene flanked by homologues of target genes was amplified by PCR and electro-transformed into E.coli MC1000.When induced by L-arabinose,the plasmid pKD46 could express three recombinant proteins of λ-prophage,which led to the replacement of target gene(qseC) with chloramphenicol-resistant gene.Then the chloramphenicol-resistant gene was eliminated by FLP-promoted recombination events.The biofilm formation of wild-type and mutant strain was detected by crystal violet staining.Results The qseC-deleted mutant of E.coli was confirmed by various PCR and DNA sequencing.Gene qseC was completely deleted.There was no significant difference in growth ability between the qseC mutant strain and the wild-type strain MC1000.The biofilm formation of wild-type and mutant strain was quantified by crystal violet staining.The absorbance determined with a plate reader at 570 nm was 1.00±0.15 and 0.47±0.10 respectively.Conclusion The qseC-deleted mutant of E.coli was constructed successfully.And the qseC gene plays an important role in regulation of biofilm formation in E.coli.

The ptrpose of this study was to investigate the effects of long-term estradiol and progesterone combined therapy on bone mineral density (BMD) in patients with Turner syndrome.Eight patients with Turner syndrome received estradiol and progesterone combined therapy for six years were observed and BMD was measured for each of them before hormone replacement therapy (HRT) and at an interval of one to two years after HRT and compared with that in normal age-sex matched women.BMD was significantly lower in patients with Turner syndrome than that in normal controls before HRT.All the patients with Turner syndrome had breast enlargement and irregular vaginal bleeding after HRT.BMD increased slightly in the patients with Turner syndrome after six-year HRT ,but still much lower than that in normal controls.Their BMD of the 2nd to 4th lumbar vertebra increased to (0.84±0.22) g/cm2 after HRT from (0.75±0.12)g/cm2 before it,with Z-score increased to -2.2±0.6 from -3.2±0.9,respectively; and overall BMD of the hip increased to (0.81±0.08) g/cm2 after HRT from (0.68±0.07) g/cm2 before it,with Z-score increased to-1.2 ± 0.3 from-2.2 ± 0.5,respectively.Long-term HRT can improve their BMD for patients with Turner syndrome but can not restore it to normal.

Objective To study the effect of Bi-level positive airway pressure (BiPAP) on hemodynamics in patients with the chronic obstructive pulmonary disease (COPD) combined coronary heart disease.Methods One hundred patients with COPD combined coronary heart disease treated by BiPAP ventilation were enrolled.The blood gas analysis and the hemodynamics were monitored and analyzed in patients with the COPD combined coronary heart disease before treatment and after BiPAP ventilation treatment for 2 hours,24 hours,72 hours and 1 week.Results PaCO2 decreased significantly after 2-hour's treatment by BiPAP ventilation( P ＜ 0.05) and the heart rate and systolic blood pressure also decreased significantly after 24-hour's treatment by BiPAP ventilation.The left ventricurlar ejection fraction( [ 65.63 ± 6.86 ] ％ vs.[ 56.21 ±5.26]％,P ＜ 0.05 )was significantly improved after BiPAP reatilation treatment for one week.The mean pulmonary arterial pressure ( [ 3.74 ± 0.96 ] vs [ 5.12 ± 1.12 ] kPa,P ＜ 0.01 ),angina pectoris ( [ 0.20 ± 0.01 ]time/d vs [ 0.69 ± 0.03 ] time/d,P ＜ 0.05 ) were significantly decreased.Conclusion COPD combined coronary heart disease patients may achieve an optimal effect by BiPAP ventilation.BiPAP ventilation has no impact on the hemodynamics in patients with the COPD combined coronary heart disease.

The mechanism of Tongmai Oral Liquid (TOL) for chronic nephritis rat model with Qi-deficiency and blood-stasis syndrome was studied.The model was established by adding Qi-deficiency and blood-stasis syndrome to the chronic nephritis rats. The experimental rats were randomly allocated to 5 groups: Group A (treated with high dosage of TOL), Group B (treated with low dosage of TOL), Group C (treated with Jinshuibao), Group D (model control group) and Group E (normal control group). After 4 weeks of treatment, general health state, biochemical indexes including T lymphocyte subgroup and blood rheology, and pathological damage of kidney tissue were much improved in Group A than Group B and Group C. It is indicated that TOL can improve the renal function and delay the occurrence of glomerular arteriosclerosis in rats by reinforcing Qi, activating blood flow, regulating immune function, lessening the hypercoagulative state and reducing renal damage.

Objective To observe serum secreted protein acidic and rich in cysteine (SPARC) levels in normal subjects,obese subjects,patients with type 2 diabetes mellitus (T2DM),and obese patients with T2DM,as well as the difference of SPARC expression in mesenteric adipose tissue of subjects with and without T2DM.Methods Serum SPARC level was measured with the ELISA.RT-PCR,Western blot,and immunofluorescence were used to examine SPARC mRNA and protein expressions in mesenteric adipose tissue.Results (1) SPARC levels were higher in obesity,T2DM,and T2DM with obesity groups compared with normal group [(1 191.6 ± 718.91,1 223.81 ± 645.96,1 538.01 ± 757.95 vs 851.07 ± 280.21) ng/L,P＜0.05].(2) Pearson correlation analysis showed that SPARC level was positively correlated with homeostasis model assessment for insulin resistance index (r =0.205,P＜ 0.05).(3) The expression levels of SPARC mRNA and protein in mesenteric adipose tissue of T2DM patients were higher than those of control subjects (P ＜ 0.05).Conclusion SPARC is closely related to the development of obesity,insulin resistance,and type 2 diabetes mellitus.

Objective To study the effect of epinephrine on biofilm formation of the qseC-deleted mutant of Escherichia coli on biomaterial.Methods The strains used in this study are Escherichia coli MC1000 and MC1000AqseC.LB was used for all the experiments.To determine the effect of epinephrine on motility,halos were measured in LB medium at 37℃ in the presence of epinephrine(50 μmol/L).LB with epinephrine and without epinephrine were used,and then the experiment of bacterial biofilm formation on PVC material was taken.The relative amount of biofilm was estimated.The thickness of bacterial community and bacterial community quantity in the unit area on PVC materials were measured by confocal laser scanning microscope( CLSM),and the surface structure of biofilm formation was observed by scanning electron microscope(SEM).Results The mutant strain formed less biofilm than the wild-type strain in LB.The increment in motility of wild-type strain due to epinephrine addition was shown,but mutant strain is unaffected.Similarly,biofilm formation of the wild-type strain was increased by epinephrine,but epinephrine did not affect the biofilm formation of the qseC mutant.The CLSM and SEM showed that epinephrine stimulated biofilm formation of wild-type strain on PVC materials,but had no effect on qseC-deleted mutant strain.Conclusion Epinephrine increases Escherichia coli biofilms on biomaterials through qseC.

Objective To investigate the role of quorum sensing Escherichia coli regulator C (qseC) in intestinal bacterial translocation in rats subjected to hemorrhagic shock.Methods Thirty Sprague-Dawley rats,weighing 250-300 g,were randomly divided into 5 groups (n =6 each):control group (group C),MC1000-sham shock group (group M-SS),MC1000qseC-sham shock group (group △-SS),MC1000-hemorrhagic shock group (group M-HS),and MC1000△ qseC-hemorrhagic shock group (group △-HS).The rats drank 150 μg/ml of disinfect water containing streptomycin in 3 consecutive days to inhibit the autochthonous flora in the intestinal tract.From 4th day,the rats were fed with Escherichia Coli MC1000 or MC1000△ qseC 1 ml/100 g by gastric perfusion once a day for another 3 consecutive days in the other 4 groups,while the rats were fed with normal saline instead in group C.Hemorrhagic shock was induced by blood-letting.The mesenteric lymph node (MLN),spleen and liver specimens were obtained at 24 h after operation for bacterial culture and the bacteria were identified.Bacterial translocation from gut to MLN,spleen and liver was observed and the number of bacteria in MLN,spleen and liver tissues were counted.Results The rate of bacterial translocation was significantly higher,and the number of bacterial colonies in MLN,spleen and liver tissues and the total number of bacterial colonies were significantly larger in groups M-HS and △-HS than in group C,and in group M-HS than in groups M-SS and △-SS (P ＜ 0.05).The rate of bacterial translocation was significantly lower,and the number of bacterial colonies in MLN,spleen and liver tissues and the total number of bacterial colonies were significantly smaller in group △-HS than in group MHS.Conclusion QseC is involved in the intestinal bacterial translocation following hemorrhagic shock in rats.

Objective To study the influence of cochlear implantation on residual hearing in children .Methods Behavioral audiometry were performed pre -implant and 3~21 months post -implant on thirty -four cochlear implant recipients with severe to profound hearing loss .According to follow -up time ,they were divided into 2 groups which were Group A(3~12 months ,21 cases) and Group B(≥13 months ,13 cases) .The thresholds at 250 Hz ,500 Hz ,1 000 Hz and 2 000 Hz were analyzed .Results There were 25 out of 34 patients (73 .53% ) had partial residual hearing after cochlear implantation .Comparing to the hearing loss pre -operation and post -operation , which were most obvious at 500 Hz ,followed by 250 Hz ,1 000 Hz ,2 000 Hz (P<0 .05) ,and there were significant different among different frequencies .There was significant difference at different frequencies at hearing loss thresh-olds only in Group A .But there was no significant difference in Group B .With the prolonged time after the cochlear implantation ,residual hearing at all frequencies showed a trend of recovery .Conclusion The residual hearing could be partial preserved after cochlear implant in pediatric patients with severe to profound hearing impaired ,the residu-al hearing at lower frequencies (250 Hz ,500 Hz) were less affected than those at higher frequencies .With the pro-longed time after the cochlear implantation ,the residual hearing showed a certain degree of recovery .

Child ; Contracture ; congenital ; therapy ; Humans ; Male ; Neck Muscles ; pathology ; Torticollis ; congenital

Objective To explore whether different Epstein-Barr virus (EBV) infection status is involved in systemic lupus erythematosus (SLE) pathogenesis through type Ⅰ interferon pathway by observing the EBV antibodies,serum interferon α (IFN-α) level and four type Ⅰ interferon induced gene (ISGs;2'-5' oligoadenylate synthetase-like protein (OASL),myxovirous resistant 1 (MX1),interferon-stimulated gene 15 (ISG15) and lymphocyte antigen 6 complex,locus E (LY6E) expressions in SLE patients and healthy controls.Methods Forty-eight patients with SLE and 36 healthy controls were enrolled into this study.The serum antibodies of EBV capsid antigen-IgG/lgM and EBV nuclear antigen 1-IgG,and serum levels of IFN-α were measured by enzyme linked immunosorbent assay (ELISA) method.Real time reverse transcription polymerase chain reaction was used to test the mRNA levels of OASL,MX1,ISG15 and LY6E in the peripheral blood lymphocytes (2-△△Ct was used to indicate the gene expression).Statistical analysis was performed using t-test or Mann-Whitney U test,Chi square test and Spearman correlation test.Results ① The EBV lytic infection rate (VCA-IgM) in SLE patients was higher than that in healthy controls (40％ vs 11％,x2=5.381,P=0.027).② The serum levels of IFN-α in SLE patients were higher than those in the healthy controls [(206±151) ng/L vs (90± 76) ng/L,t=4.248,P＜0.05],as well as the mRNA levels of OASL,MX1,ISG15 and LY6E [1.8(0.6,5.1) vs 1.2 (0.5,1.4);1.9(1.0,4.4) vs 0.9(0.7,2.5);4.1(1.6,7.8) vs 0.8(0.5,1.7);1.6(0.7,3.3) vs 0.8(0.6,1.2),U=604,560,312,608;P＜0.05,respectively].The mRNA levels of OASL,MX1,LY6E and ISG15 were all positively related to SLE disease activity index (SLEDAI) scores (r=0.319,0.461,0.547,0.484,P＜0.05,respectively).Serum IFN-α levels were elevated in SLE patients with EBV lytic infection than in non-lytic infection patients [(282± 174) ng/L vs (157±114) ng/L,t=2.604,P＜0.05];The mRNA levels of OASL and ISG15 were also elevated in patients with lytic EBV infection [2.0(0.8,7.6) vs 1.2(0.6,3.1);6.2(2.4,15.5) vs 3.3(1.3,6.3),U=377,350,385,354;P＜0.05,respectively].The SLEDAI scores in patients with EBV lyric infection were higher than in patients with non-lyric infection (16±4 vs 12±8,P＜0.05).Conclusion EBV infection may be involved in the pathogenesis of SLE by activating the type Ⅰ interferon pathway.