Objective To construct the serine protease gene(sprE)mutant and to study the pathogenicity of sprE gene of Enterococcus faecalis.Methods Recombinant suicide vector pCQ001 of Enterococcus faecalis with pTX4577,was constructed.Then,created isogenic sprE-deficient mu- tant(*sprE)by allelic replacement was constructed.Moreover,the growth ability and the virulence of the mutant were compared with those of the wide type in vitro and in vivo respectively.A mouse peritonitis model and a rabbit endocarditis model were utilized in the study.Results The *sprE was selected by kanamycin and identified by polymerase chain reaction(PCR),pulsed field gel electropho- resis(PFGE)and Southern blot.The evidences showed that the sprE gene had a major role in helping bacteria to resist the elevated temperature and oxidative stress.The virulence of mutant decreased af- ter sprE gene was knocked out.Conclusions The *sprE of Enterococcus faecalis is constructed suc- cessfully,sprE gene is important in the pathogenesis of Enterococcus faecalis,which probably is a major virulence factor of Enterococcus faecali.

[Objective]To explore the intracellular antimicrobial activities of erythromycin,ciprofloxacin,levofloxacin,moxifloxacin against Legionella pneumophila.[Methods]The minimum inhibition concentration(MIC)of each antibiotic was evaluated by E-test method and microdilution method respectively.The minimal extracellular concentration inhibiting intracellular multiplication(MIEC)of each antibiotic was evaluated by the MTT colorimetric assay system.[Results]The MIC concentration for each drug by E-test method were:erythromycin,0.047 μg/mL;ciprofloxacin,0.38 μg/mL;levofloxacin,0.125 μg/mL;moxifloxacin,0.125 μg/mL,the MIC concentrations for each drug by microdilution method were:erythromycin,0.125 μg/mL;ciprofloxacin,0.03 μg/mL;levofloxacin,0.016 μg/mL;moxifloxacin,0.016 μg/mL.The MIEC concentration for each drug were:erythromycin,0.25 μg/mL;ciprofloxacin,0.016 μg/mL;levofloxacin,0.016 μg/mL;moxifloxacin,0.004 μg/mL.[Conclusions]Fluoroquinolones have superior activity than erythromycin in U937 cells infected with L.pneumophila.Moxifloxacin is the most potent drug among the four tested antimicrobials.Our results indicated that the MTT assay system allows comparative and quantitative evaluations of the intracellular activities of antibiotics against L,pneumophila and efficient processing of a large number of samples.

Objective To construct a recombinant expressing plasmid of the hy1 gene of Enterococcus faecium and to express the recombinant Hy1 protein in E.coil.To explore the immune response in mice fed orally with Hyl protein.Methods hy1 gene was amplified by polymerase chain reaction(PCR)and inserted into a prokaryotic expression vector pQE-30.The recomhinant plasmids were transfected into DH5_?to express Hy1 fusion proteins,which were purified by Ni~--column. Western blot was employed to confirm the immunogenicity of the purified protein.Mice were immu- nized by feeding with the fusion protein.The concentrations of antigen-specific antibody in the serum, mucosal fluid and faces were detected by enzyme-linked immunosorbent assay(ELISA).The role of these antibodies in the anti-infection response was evaluated after the mice were challenged with TX0016.Results hy1 gene was sequenced as 1662 bp,the fusion protein encoding polypeptides of 553 amino acid residues.The relative molecular weight was 60 000 when it was determined by sodium dodecylsulfatepo-lyacry-lamide gel electropboresis(SDS-PAGE).The dissolvable expression protein accounted for 38% of total cell protein.After processed by affinity chromatography,the purity of fusion protein was above 92%.Western blot analysis confirmed that fusion protein could be specifically recognized by the anti-TX0016 serum.The concentrations of serum IgA,serum IgG,faeces sIgA and intestmucosal fluid sIgA was 0.365?0.048,0.431?0.064,0.743?0.056 and 1.112?0.113 respectively in hy1 groups and 0.051?0.013,0.098?0.019,0.102?0.032 and 0.187?0.051 respectively in control group.The differences were statistically significant.The mice survival rate after TX0016 challenge was 70% in hyl group and 50% in control group.There was significant difference between these two groups.Conclusion The results indicate that oral immunization with hyl can induce effective mueosal immune response and produce high level sIgA.

Adult ; Burkitt Lymphoma ; metabolism ; pathology ; CD3 Complex ; metabolism ; DNA Nucleotidylexotransferase ; metabolism ; Diagnosis, Differential ; Female ; Granulosa Cell Tumor ; metabolism ; pathology ; Humans ; Ki-67 Antigen ; metabolism ; Leukocyte Common Antigens ; metabolism ; Ovarian Neoplasms ; metabolism ; pathology ; surgery ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ; metabolism ; pathology ; surgery ; Sarcoma, Myeloid ; metabolism ; pathology

AlM: To investigate the association between the degree of anterioruveitis and related factors including inflammatory markers as well as sacroiliac joint imaging in patients with ankylosing spondylitis ( AS) .
METHODS: Anterior changes evaluated by slit lamp, erythrocyte sedimentation rate ( ESR ) , C - reactive protein ( CRP ) and magnetic resonance imaging of 55 cases with AS associateduveitis were retrospectively analyzed. A modified endotoxin-induced uveitis ( ElU ) clinical standard was used for uveitis grading. SPARCC sacroiliac scoring was used to evaluate bone edema of sacroiliac joint. The correlation between the degree of uveitis and sacroiliitis was assessed.
RESULTS: ln the 55 patients with AS, ElU grading scored 2-10, and SPARCC index scored 0-22. Further analysis showed that the severity of uveitis was significantly correlated with ESR (r=0. 869, P<0. 001) and CRP (r=0. 485, P<0. 001). The degree of anterior uveitis in AS patients was not correlated with inflammation of sacroiliac joint (r=0. 237, P=0. 081).
CONCLUSlON: Local autoimmunity of uveitis and sacroiliac joint inflammation with subsequent bone formation in AS might be mutually independent processes.

Objective To study the relationship between environmental chemical elements,vires infection and dilated cardiomyopathy(DCM).Methods In 2008,233 patients with DCM(case group)and 150 patient with stable angina(control group)were chosen in Liaocheng People's Hospital and Yanggu People's Hospital,Shandong province.Population distribution and disease history were surveyed in the two groups.Human myocardial antibody IgG(AMA-IgG),Coxsackie B virus IgG(CBV-IgG),Adenofirus antibody IgG(ADV-IgG)were detected by ELISA in both the case group and the control group.Serum trace elements were detected in the two groups.The general chemical and toxicological indicators in drinking water of the high-and the low-incidence aireas of the disease were control group[60.00%(90/150),χ2=13.80,P<0.01)].Per capita annual income(Yuan,RMB)in the case group (3207.82±618.51)was lower than that of the control group[(5086.61±886.12),t=24.40,P<0.01].Personal alcohol consumption in the case group[(175.00±160.50)g/d]was higher than that of the control group[(110.22±100.03)g/d,t=4.40,P<0.01)].The rate of myocarditis in the case group[5.15%(12/233)]was higher than ADV-IgG in the cage group were 7.78%(7/90),6.67%(6/90)and 6.67%(6/90),respectively.Compared with those in the control group[3.33%(2/60),5.00%(3/60)and 5.00%(3/60),χ2=1.26,0.18,0.18,all P>0.05],no mg/L]in drinking water of the high-incidence areas were significantly higher than that of iron[(0.39±0.67)mg/L,t=2.11,P<0.05]and that of manganese[(0.15±0.14)mg/L,t=3.01,P<0.01]in the low-incidence arefas.The content of semm iron[(69.1±57.8)μmol/L]in the case group evidently exceeded the normal range(15.6-35.9 μmol/L)and obviously higher than that in the control group[(20.0±17.5)μmol/L,t=5.04,P<0.01].Conclusions Theso data do not support that DCM is related with persistent virus infection and autoimmunization.DCM is probably related with low incomes,high alcohol consumption,myocarditis,high iron and manganese contents in drinking water and high content of serum iron.

Objective To research the morphological classification of inferior angle of scapula based on CT 3D reconstruction and its clinical significance. Methods A total of 290 scapular bones data were collected from People's Hospital of Lu Xian County and measured based on CT 3D reconstruction. The scapular bones images were reconstructed by CT 3D technique. Reference points; g was the inferior scapular angle, n was the inferior glenotubular nodule, m was the intersection of the scapular ganglion and the medial edge of the scapula, k was the upper scapular angle, r was the notch of the scapular glenoid, h was the intersection of point g to mr. The parameters of inferior angles of 290 scapular bones, including the thickness of point g (the thickest part of inferior angle of the scapula), the length of the line gn, line gm, and line gh, and the angle of ∠ ngm, ∠ gmk (the projection of the scapular coronal plane), and ∠ gmk' (the projection of the scapular sagittal plane) were observed and measured by CT 3D technique. Results Under the CT three-dimensional reconstruction, the inferior angle of scapula was classified into three types; accessory angle-type ( 175, 60. 34%), U-type ( 81, 27. 93%) and V-type (34, 11. 73%). In addition, there was a significant difference between the auxiliary angle type and the V type on the thickest part of the g point and gn (P<0.05); U-type and V-type with paragonai gm-significant difference (P<0.05); There was a significant difference between the auxiliary angle type and the U shape on gh (P<0.05); there was a significant difference between the auxiliary angle type and the U shape on ∠ gmk' (P<0.05). There was a significant difference in the inferior angle of the bilateral scapula on mngm (P<0.05). Conclusion Based on the result of CT three-dimensional reconstruction, the inferior angle of scapulae have three anatomical types, with the accessory angle-type as the main type, and the morphology and classification of it are of certain clinical implication.

OBJECTIVETo explore the effects of n-hexane on expression of serum myelin proteins (MBP) in workers occupationally exposed to n-hexane.

METHODSIn this study, 269 workers exposed to n-hexane for more than one year and 104 subjects not exposed to n-hexane served as the exposure group and the control group, respectively. The urinary 2,5-hexanedione levels in all subjects were detected. On the basis of urinary 2,5-hexanedione levels, the exposure group was divided into the high exposure sub-group and low exposure sub-group. The serum myelin basic protein (MBP) levels were measured by ELISA kit.

RESULTSThe mean concentration of urinary 2,5-hexanedione in the exposed group was (3.10 +/- 1.35) mg/L. The concentration of urinary 2,5-hexanedione in the control group was undetectable. The levels of serum MBP in the high exposure sub-group and low exposure sub-group were (2.43 +/- 0.24) and (1.62 +/- 0.23) microg/L, respectively, which were significantly higher than that (0.78 +/- 0.12) microg/L in the controls (P < 0.01). Pearson correlation analysis showed the positive correlation between serum MBP levels and urinary 2,5-hexanedione levels (r = 0.781, P < 0.01).

CONCLUSIONThe results of present study showed that the serum MBP levels of workers occupationally exposed to n-hexane significantly elevated, and the serum MBP can serve as the effective biomarker of n-hexane exposure.

Adolescent ; Adult ; Case-Control Studies ; Female ; Hexanes ; adverse effects ; Humans ; Male ; Myelin Basic Protein ; blood ; Occupational Exposure ; Young Adult

Objective To investigate the clinical effect of noninvasive mechanical ventilation in the treatment of chronic obstructive pulmonary disease (COPD) and the expression of insulin-like growth factor (IGF) Ⅰ, Ⅱand cathepsin B (CB) in patients.Methods 89 patients with chronic obstructive pulmonary disease from January2014 to June 2017 in the hospital for treatment were retrospectively chosen as the research objects. They were randomly divided into the control group (n = 43) and the study group (n = 46). The control group was treated with conventional therapy. The study group was treated with noninvasive mechanical ventilation on the basis of the control group. The observation period was 10 days of treatment, the level of the indicators of lung function, arterial blood gas changes of the both groups were detected, and the clinical treatment efficiency was taken for statistics. At the same time, the levels of IGF-Ⅰ, Ⅱ and CB in patients were analyzed.Results (1) The lung function of the study group: the lung function VC [ (302.72 ± 40.79) m L], the maximum expiratory flow PEF (421.72 ± 37.59) L/min], and the first expiratory volume FEV1 [ (83.24 ± 11.65) ％], were significantly higher than that of the control group (P＜0.05). (2) The arterial blood gas in the study group: arterial oxygen partial pressure (PaO2) [ (99.24 ± 10.16) mm Hg], partial pressure of carbon dioxide (PaCO2) [ (43.32 ± 5.37) mm Hg], oxygenation index (PaO2/FiO2) [ (281.02 ± 19.74) ％ ], were better than the level of detection in the control group (P＜0.05). (3) The effective rate of clinical treatment was 93.48％ in the study group, which was significantly higher than that in the control group of 67.44％ (P＜0.05). (4) The IGF-Ⅰ [ (11.47 ± 1.53) ug/L] and IGF-Ⅱ [ (28.27± 11.49) ug/L] in the study group were lower than those in the control group, while CB [ (1.03 ± 0.24) ng/ml]was higher than that of the control group (P＜0.05).Conclusions Noninvasive mechanical ventilation is helpful to clear the recovery of lung function in patients with chronic obstructive pulmonary disease, can alter the abnormal expression of insulin-like growth factor I, II and cathepsin B levels in patients, and the clinical treatment effect is significant.

The purpose of the present study was to explore the role of endogenous hydrogen sulfide (H2S) in pulmonary arterial hypertension induced by endotoxin. Adult male Sprague-Dawley (SD) rats were randomly divided into four groups: Control group (0.5 mL/kg body weight of normal saline, i.v.), lipopolysaccharide (LPS)-treated group (5 mg/kg body weight of LPS, i.v.), LPS + NaHS (5 mg/kg body weight of LPS, i.v., and 28 μmol/kg body weight of NaHS, i.p.) and LPS + PPG group (5 mg/kg body weight of LPS, i.v., and 30 μmol/kg body weight of PPG, i.p.). Rats were anesthetized with 20% urethane (1 g/kg body weight, i.p.). A polyethylene catheter was inserted into the pulmonary artery through the right external jugular vein to measure the mean pulmonary arterial pressure (mPAP) for 7 h, and then the pulmonary artery was isolated rapidly by the method described previously. Pulmonary arterial activity was detected. H2S concentration and cystathionine γ-lyase (CSE) activity in pulmonary artery tissues were determined by biochemical method. CSE mRNA expression was detected by competitive reverse transcriptase-polymerase chain reaction (RT-PCR). Compared with control, LPS significantly increased mPAP [(1.82±0.29) kPa vs (1.43±0.26) kPa, P<0.01], decreased H2S production [(26.33±7.84) vs (42.92±8.73) pmol/g wet tissue per minute, P<0.01), and reduced endothelium-dependent relaxation response [(75.72±7.22)% vs (86.40±4.40) %, P<0.01) induced by ACh (1×10(-6) mol/L). These effects were partly reversed by co-administration of NaHS and enhanced by co-administration of PPG. Both CSE activity and CSE mRNA expression were consistent with H2S production. It is suggested that the inhibitory effect of LPS on endothelium-dependent relaxation results in pulmonary hypertension, which might be mediated through H(2)S.
Animals ; Arterial Pressure ; Cystathionine gamma-Lyase ; metabolism ; Hydrogen Sulfide ; metabolism ; Hypertension, Pulmonary ; chemically induced ; metabolism ; Lipopolysaccharides ; adverse effects ; Male ; Rats ; Rats, Sprague-Dawley ; Sulfides ; pharmacology