Objective　To explore the possibility of R plasmid (pRST98) encoding resistance to antimicrobial agents in S. typhi mediate virulence to its host bacteria. Methods　pRST98 was transferred into a less virulent strain of S. typhimurium RIA for creating a transconjugant pRST98/RIA. The standard S. typhimurium virulence strain SR-11 which carries a 100kb virulence plasmid was used as a positive control, and RIA as a negative one. Infection with S. typhimurium was employed to assess the effect of pRST98 on the virulence of host bacteria by LD50 of peroral (p.o.) and intraperitoneal (i.p.) infection of BALB/c mice. After oral inoculation of the bacteria into BALB/c mice, mesenteric lymph nodes, spleen and liver were examined for quantity of bacteria and for the histopathological changes. The three strains were also studied for their in vitro adhesion and invasion of HEp-2, CHO and HeLa cells. Three chromosomally isogenic strains, i.e., plasmid-containing wild-type S. typhi strain, plasmid artificially cured strain and plasmid-reintroduced into the cured strain were compared for their resistance to the bactericidal activity present in human, rabbit and guinea pig sera. Results　The p.o. and i.p. LD50 of pRST98/RIA was 700 and 75 folds less than that of RIA. The bacteria counting in mesenteric lymph nodes, spleen and liver were more (P<0.05) and more severe histopathological changes were observed in pRST98 harboring S. typhimurium. However, pRST98 did not affect the adhesion and invasion of S. typhimurium to the cells. Experiments showed that pRST98 mediated resistance to serum bactericidal effect in host bacteria (P<0.05). Conclusion　This is the first report about a plasmid carrying genes coding for drug resistance and virulence in S. typhi. The data presented here suggested that novel virulence gene or related sequences, which are important in the pathogenesis of Salmonella infection could exist on pRST98.

Objective To assess the dagnostic value of computed tomography(CT) for recurrent carcinoma of larynx after operation. Methods 45 CT cases of recurrent carcinoma of larynx were analysed retrospective and were compared with pathology results.Results CT was capable of demonstrating the existence,border and range of the recurred tumor and showing the information about inetastasis in lymph node of neck,the involved throat space and the invaded blood vcsscls in cervical.Conclusion CT is an cxact,rcliable and convenient method to evaluate the recurrence of larynx carcinoma,and is helpful to make therapy planning.

Objective To investigate the relenvant factors of precocious puberty and provide the basis for precaution of high risk group of precocious puberty .Methods A 1:1 case-control study was performed on 50 healthy girls and 50 precocious puberty girls .Results Univariate analysis showed an significant differences in the exposure rates of following five factors between cases and controls:intake of nutritious and health products ,favoring meat and poultry,long time watching television,addiction to TV ads,family economic condition (The Chi-square values are 7.045,9.073,17.478,14.063,12.246 respectively,all P<0.05).The Logisitc regression showed that favoring meat and poultry,long time watching television and addiction to TV ads had significant differences among precocious puber -ty children(The Wald′s Value were 4.846,6.850,5.662 respectively,all P<0.05).Conclusion favoring meat and poultry,long time watching television and addiction to TV play an important roles in precocious puberty .

Objective Matrix metalloproteinase-10 (MMP-10) has been shown to be highly associated with atherosclerosis.Recent studies showed that levels of MMP-10 were elevated in infarcted tissues in acute ischemic stoke.However,serum levels of MMP-10 in patients with acute ischemic stroke have never been studied previously.This study aims to investigate the serum levels of MMP-10 in patients with acute ischemic stroke,and evaluate the association of serum levels of MMP-10 with stroke subtypes based on Trial of Org 10 172 in acute stroke treatment classifications,the severity of stroke,risk factors and carotid artery plaque.Methods The circulating levels of MMP-10 were measured by enzyme linked immunosorbent assay in 194 subjects,including 109 patients who were diagnosed as acute ischemic stroke in the Department of Neurology,Shengjing Hospital,China Medical University from April to December 2012,and the 85 healthy controls.Results Patients with acute ischemic stroke had higher serum levels of MMP-10 compared with healthy controls (6.59 (6.07,7.31) μg/L vs 5.16 (3.87,5.94) μg/L,Z =8.33,P ＜ 0.01).NIHSS score had positive correlation with serum levels of MMP-10 (r =0.204,P =0.037).Classified by risk factors,we compared the MMP-10 levels of subsets,and results displayed that statistically significant difference existed between dyslipidemia subset and non-dyslipidemia subset (Z =2.07,P =0.042).MMP-10 levels had positive correlation with serum levels of LDL-cholesterol (r =0.248,P =0.040),but negative correlation with thrombin-activatable fibrinolysis inhibitor (TAFI;r =-0.208,P =0.030).The subset with unstable plaques had higher MMP-10 levels than that with stable plaque (6.62 (6.13,7.36) μg/L) vs 6.10 (6.00,6.46) μg/L,Z =2.12,P =0.034),implying the relationship of MMP-10 and atherosclerosis.Conclusions Patients with acute ischemic stroke have higher serum levels of MMP-10 compared with the healthy controls,and MMP-10 levels have positive correlation with the severity of stroke.MMP-10 is associated with the subtypes of stroke classified by risk factors,and dyslipidemia subset has higher levels of MMP-10 than that of non-dyslipidemia subset.MMP-10 has positive correlation with LDL-cholesterol,but negative correlation with TAFI.MMP-10 may be involved in the process of formation and disruption of unstable plaques,which contribute to the stenosis of arteries and onset of acute ischemic stroke.

BACKGROUND: Denddtic cells (DCs) constitute the dominant population of antigen presenting cells (APCs) by possessing potent ability to initiate T cell immunity. The ultrastructure study of DCs is less reported. OBJECTIVE: To investigate the ultrastructure of DCs from mice bone marrow at different maturation stages, and the morphology of DCs between CD40 ligation and tumor necrosis factor-alpha (TNF-α) stimulation in vitro. METHODS: Mice myeloid DCs were generated from bone marrow in vitro using granulocyte-macrophage colony-stimulating factor (GM-CSF)and interleukin-4 (IL-4). Immature DCs were loaded with apoptotic tumor cells (AP-DC), and AP-DC was then stimulated with CD40L-CHO cells and TNF-α for 48 hours, respectively. DCs were routinely sectioned, and ultrastructure was observed under transmission electron microscope. RESULTS AND CONCLUSION: Immature DCs showed a few short and blunt cytoplasmic processes, there were specific morphology lysosomes that liked earphone in some cells; DCs engulfing the apoptotic bodies were observed; sub-cellular structures between CD40 ligation and TNF-α stimulated DCs were different, the former had typical morphology of mature DCs which exhibited many dendritic protrusions, however, some DCs displayed apoptosis and autophagy after TNF-α stimulation. In a conclusion, CD40 ligation plays an essential role in myeloid DCs differentiation and maturation, TNF-α can mediate apoptosis and autophaqy of DCs.

Objective:To explore the efficacy of group cognitive-behavioral therapy (GCBT) to generalized anxiety disorder (GAD).Methods:In this randomized controlled trial,the control group (n =23) and the intervention group (n =33) were included,all of the participants received duloxetine (30-120 mg/d) as pharmacotherapy.The intervention group received 8 group cognitive-behavioral therapy sessions weekly,90 minutes for each time.Assessments were conducted with the Hamilton Anxiety Scale (HAMA),Hamilton Depression Scale (HMAD) and Clinical Global Impression (CGI) at baseline,mid-treatmentand post-treatment.CGI included three factors,the severity of illness (SI),the globalimprovement (GI) and the efficacy index (EI).Results:The repeated measures analysis of variance of HAMA showed that,there were statistical significance on the HAMA for interaction between measure time and group processing (F =4.35,P ＜ 0.05).Compared with the control group,the intervention group got higher decreased scores of HAMA at the 4th week and 8th week,and higher prevalence of being cured and efficient at the 4th week.At the 8th week,the decreased scores of HMAD were higher in the intervention group than in the control group,and the scores of CGI-SI and the CGI-IE were lower in the intervention group.Conclusion:It suggests that group cognitive-behavioral therapy combined with pharmacotherapy could be earlier to be effective,and the symptoms of anxiety,depression and the state of illness could be improved more significantly compared with pharmacotherapy alone.

BACKGROUND:Previous studies discovered that pRST98, original y isolated from Salmonel a enterica serovar typhimurium (S.typhimurium) could promote bacterial biofilm formation. In addition, bacterial harboring pRST98 can promote the secretion and expression of interleukin-10 after infection in cells and animals.
OBJECTIVE:In vitro studies have discovered the effects of interleukin-10 at varying concentrations and conjugative plasmid pRST98 on the biofilm formation of S.typhimurium.
METHODS:S.typhimurium wild-type strainχ3306, virulence plasmid-deletion S.typhimurium strainχ3337 and pRST98-transconjugant S.typhimuriumχ3337/pRST98 were established in vitro and cultured for biofilm formation. 1, 10, 100 μg/L interleukin-10 were added during the biofilm formation. 0 μg/L interleukin-10 was set as a control. Crystal violet staining method, semi-quantitative method, confocal laser scanning microscopy and scanning electron microscopy were used to determine the effects of interleukin-10 on the biofilm formation and compare the effects of S.typhimurium with or without pRST98.
RESULTS AND CONCLUSION:Intra-group comparison showed that, compared with the control group, S.typhimurium gathered together and formed thicker biofilm in concentration of 1 and 10 μg/L of interleukin-10. The promotion effects of S.typhimurium on biofilm formation were greatly improved in 10 μg/L. Interleukin-10 in 100 μg/L inhibited S.typhimurium biofilm formation. Inter-group comparison showed that, A570 inχ3337/pRST98 was greatly higher than that inχ3306 andχ3337 under the same concentration of interleukin-10. The results indicate that both 1 and 10 μg/L of interleukin-10 promote biofilm formation, especial y bacteria harboring pRST98.

OBJECTIVE To investigate the protective effect of curcumin on Aβ1-42 damaged cells. METHODS SH-SY5Y cells were cultured with Aβ1-4210μmol · L-1 in the absence or presence of curcumin 1, 5 or 10 μmol · L-1. Cell viability was assayed by MTT. Cell membrane damage was detected by the concentration of lactate dehydrogenase (LDH) in culture medium. Cell apoptosis was measured by flow cytometry with Annexin Ⅴ-FITC/PI staining. Mitochondrial membrane potential was characterized by fluorescence of JC-1 dye. Enzymatic activity of caspases-9 and-3 was measured by colorimetric assay. Protein expression of caspase-3 was detected by Western blotting. RESULTS Compared with vehicle control, the cell viability, concentration of LDH and both early and late apoptosis in Aβ1-4210 μmol · L-1 damaged group were decreased(P<0.01). However, the cell viability, release of LDH and both early and late apoptosis in curcumin group were promoted compared with that in Aβ1-4210μmol·L-1 damaged group. Curcumin inhibited Aβ1-42-induced depolarization of mitochondrial membrane potential(P<0.01), and attenuated Aβ1-42-induced activation of both caspases9 and caspases3 in a concentration-dependent manner, respectively(r=0.990, P<0.01; r=0.996, P<0.01). There were no significant differences in the above detected indexes between curcumin 10 μmol · L-1 group and vehicle control group. CONCLUSION Curcumin inhibits Aβ1-42-induced cell damage and apoptosis by promoting mitochondrial membrane potential and depressing the activation of caspases.

T cell acute lymphoblastic leukemia (T-ALL) is an aggressive leukemia. However the poor prognosis and low morbidity restrict further analysis of the disease. Therefore there is an increasing demand to develop animal models for identifying novel therapeutic approaches. In this study, we inoculated the anti-mouse CD122 monoclonal antibody conditioned NOD/SCID mice with the leukemia cells from 9 T-ALL patients and 1 cell line via the tail vein. Four of the 9 patients and the cell line were successfully engrafted. Flow cytometry detected high percentage of human CD45(+) cells in recipient mice. Immunohistochemistry showed infiltration of human CD45(+) cells in different organs. Serial transplantation was also achieved. In vivo drug treatment showed that dexamethasone could extend survival, which was consistent with clinical observation. These results demonstrated that we successfully established 5 xenotransplantation models of T-ALL in anti-mCD122 mAb conditioned NOD/SCID mice, which recapitulated the characteristics of original disease.

Objective To explore the value of static and dynamic MRI before and after operation of pelvic organ prolapse (POP). Methods 29 patients with POP (POP group)and 12 normal women (control group)underwent static and dynamic MRI.The morphologic changes of pelvic floor were observed on MR images.The measurements of bladder,uterus,Douglas pouch to pubococcygeal line (B-PCL,U-PCL,D-PCL),the puborectal hiatus line (H-line),muscular pelvic floor descent (M-line),the levator hiatus size (LHS),the levator plate angle (LPA),the iliococcygeus angle (ICA)and the urethral inclination angle (UA)were recorded on dynamic MR images.Results 19 cystoceles,28 uterine prolapses,4 rectoceles and 14 hernias of Douglas pouch were detected with MRI.29 cases of pelvic floor relaxation,27 cases of levator ani muscle defect and 24 cases of pubocervical fascial defect were found.The values of B-PCL,U-PCL, D-PCL,H-line,M-line,LHS,LPA,ICA and UA of POP group were larger than control group (P<0.01).The positions of pelvic organ returned to normal in 9 cases of 21 postoperative cases,while 12 cases remained prolapses.There was no displacement of mesh in 8 cases of mesh implant.The values of B-PCL,U-PCL,D-PCL,UA after operation were smaller than those before operation (P<0.05).Conclusion Static and dynamic MRI can evaluate morphological and functional changes of pelvic floor before and after operation of POP comprehensively,and may reveal those invisible pelvic floor dysfunction and postoperative remnant defects.