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 investigate the effects of heart displacement on hemodynamics during off-pump coronary artery bypass grafting (OP-CABG) while the sites for anastomosis were being exposed. Methods Forty-seven patients of both sexes (36 male, 11 female) aged 50-82 years undergoing OP-CABG were enrolled in the study. Preoperative cardiac function was assessed : class Ⅱ in 22 patients; Ⅲ in 23 and Ⅳ in 2 according to NYHA classification.The mean ejection fraction was 0.55?0.14 before surgery.They received on average 3.2 grafts. Premedication consisted of intramuscular morphine 10 mg, midazolam 3-5 mg and scopolamine 0.3 mg.Before induction of anesthesia ECG and SpO2 were monitored and radial artery was cannulated for continuous direct BP monitoring. Anesthesia was induced with midazolam 0.1 nig?kg-1 , fentanyl 4?g?kg-1 and pancuronium 0.1 mg?g-1 iv.The patients were mechanically ventilated after tracheal intubation and PETCO2 was maintained at about 40 mm Hg. Anesthesia was maintained with isoflurane and 50%-60% N2O in O2 and intermittent intravenous boluses of fentanyl and pancuronium. Swan-Ganz catheter which can continuously monitor mixed venous blood O2 saturation (SvO2) was placed in pulmonary artery via right internal jugular vein. SvO2, cardiac output (CO), BP, pulmonary arterial pressure (PAP) and HR were continuously monitored. Right atrial pressure (RAP) and PAWP were measured intermittently. Cardiac index (CI),stroke index (SI),systemic vascular resistance index (SVRI),PVRI, left and right ventricular work index (LVWI,RVWI) and left and right ventricular stroke work index (LVSWI,RVSWI) were calculated. The hemodynamic parameters were recorded after induction of anesthesia before surgery (T1,baseline),before heart displacement (T2), while anastomosis to anterior descending branch was being made (T3), while anastonosis to right coronary artery or posterior descending branch (T4) and to left circumflex artery or diagonal branch (T5) was being made, after normal heart position was resumed (T6) and at the end of operation (T7). Results While anastomosis to the anterior descending branch was being made (T3) SI and LVSWI significantly decreased as compared with the baseline (P

Myeloid-derived suppressor cells (MDSCs) play a crucial role in T cell dysfunction, which is related to poor outcome in patients with severe trauma. Cyclooxygenase-2 (Cox-2) contributes to immune disorder in trauma and infection via production of prostaglandin E2. However, the role of Cox-2 in the accumulation and function of MDSCs after traumatic stress has not been fully elucidated. In the present study, we treated murine trauma model with NS398, a selective Cox-2 inhibitor. Then the percentages of CD11b+/Gr-1+ cells, proliferation and apoptosis of CD4+ T cells were determined. Arginase activity and arginase-1 (Arg-1) protein expression of splenic CD11b+/Gr-1+ cells, and delayed-type hypersensitivity (DTH) response were analyzed. The results showed that Cox-2 blockade significantly decreased the percentages of CD11b+/Gr-1+ cells in the spleen and bone marrow 48 and 72 h after traumatic stress. NS398 inhibited arginase activity and down-regulated the Arg-1 expression of splenic CD11b+/Gr-1+ cells. Moreover, NS398 could promote proliferation and inhibit apoptosis of CD4+ T cells. It also restored DTH response of traumatic mice. Taken together, our data revealed that Cox-2 might play a pivotal role in the accumulation and function of MDSC after traumatic stress.

Adult ; Angiomyolipoma ; diagnostic imaging ; metabolism ; pathology ; surgery ; Antigens, Neoplasm ; metabolism ; Epithelioid Cells ; pathology ; Female ; Humans ; Kidney Neoplasms ; diagnostic imaging ; metabolism ; pathology ; surgery ; Lymphatic Metastasis ; Melanoma-Specific Antigens ; Neoplasm Proteins ; metabolism ; Nephrectomy ; Tomography, X-Ray Computed

OBJECTIVETo study the relationship between TMPRSS2: ERG gene fusion and the pathological grade of prostate cancer (PCa).

METHODSWe collected fresh prostatic tissue samples from 62 patients with PCa and another 10 with benign prostatic hyperplasia ( BPH) and included 9 cancer cell strains as the control. We examined the TMPRSS2:ERG fusion gene in the PCa samples by nest RT-PCR, compared the Gleason scores between the TMPRSS2:ERG-positive and -negative cases, and analyzed the association of TMPRSS2: ERG fusion with the pathological features of PCa.

RESULTSThe TMPRSS2: ERG fusion gene was detected in 28 (45.16%) of the PCa cases, but in none of the 10 BPH cases or the 9 cancer cell strains. No statistically significant differences were found in the Gleason scores between the TMPRSS2:ERG-positive and -negative cases (Z = -0.609, P = 0.542), but the primary Gleason score was markedly higher in the former than in the latter (Z = -2.600, P = 0.009). Univariate logistic regression analysis showed that TMPRSS2:ERG was associated with the cribriform growth pattern (OR = 6.250, P = 0.002), foamy gland morphology (OR = 6.666, P = 0.023), and signet-ring cells (OR = 3.240, P = 0.035), but multivariate logistic regression analysis manifested that it was associated with the cribriform growth pattern only (OR = 3.750, P = 0.033).

CONCLUSIONTMPRSS2:ERG gene fusion was associated with higher pathological grades of prostate cancer.

Gene Fusion ; Humans ; Male ; Oncogene Proteins, Fusion ; genetics ; Prostatic Hyperplasia ; genetics ; Prostatic Neoplasms ; genetics ; pathology

BACKGROUND:Macrophage migration inhibitory factor is involved in the process of a variety of diseases, and plays a very important role in the tumor, autoimmune diseases, inflammation, angiogenesis, fibrotic diseases and so on. These biological characteristics are similar to keloids. OBJECTIVE: To compare the distribution and number of macrophage migration inhibitory factor in normal skin, hypertrophic scar and keloid. METHODS: We colected 40 clinical pathological scar specimens after surgery, including 20 hypertrophic scars and 20 keloids. Another 10 samples of the normal skin were used as control group. Hematoxylin-eosin staining and immunohistochemistry staining were performed to test the expression of macrophage migration inhibitory factor in pathological scars and normal skin. RESULTS AND CONCLUSION:Macrophage migration inhibitory factor was positively expressed in the normal skin, hypertrophic scar and keloid, and the expression of macrophage migration inhibitory factor in keloid was significantly higher than that in hypertrophic scar and normal skin (P < 0.01). It means that the abnormal infiltration of macrophage migration inhibitory factor may be associated with the formation of keloid.

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.

Objective To compare the postoperative complications rate of total mesorectal excision(TME) performed by laparoscopic and conventional laparotomy method in treatment of rectal cancer.Methods The clinical data of patients with rectal cancer treated by laparoscopic surgery(n=42) or traditional laparotomy(n=50) were retrospectively analysed to analysis of the cause and occurrence rate of complications.(Results) The laparoscopic group had an advantage over the conventional laparotomy group in bowel movement recovery,earlier mobility and short hospital stay time.There was no difference in intraoperative injury,(anastomotic) leakage,sexual function preservation,long-term urinary dysfunction,and local recurrence or(incision) metastasis rates.Conclusions Laparoscopic TME treatment of rectal cancer had the same therapeutic effect as the conventional laparotomy procedure did,and has a better subjective outcome.

AIM: To study the inhibitory effects of 10-23 deoxyribozyme (10-23 DRz) on Escherichia coli ?-lactamase gene expression. METHODS: According to the gene sequence of Escherichia coli ?-lactamase gene blashv-5, 10-23 DRz and antisense oligonucleotides (As-ODN) were designed and synthesized. 10-23 DRz, As-ODN or control oligonucleotides were respectively introduced into Escherichia coli by the method of electroporation. Following electroporation, bacterial viability in liquid medium contained ceftazidime was detected, bacterial ?-lactamase expression was analysed by using IEF-PAGE and the ?-lactamase band was measured with gel documentation-analyzing system. RESULTS: A_ 600 in 10-23 DRz transfected Escherichia coli was lower compared with that in As-ODN transfected Escherichia coli (P

Objective To investigate the therapeutic effect of large dose mucosolvan combined variant flow rate continuous positive airway by nasal mask on neonatal respiratory distress syndrome(NRDS).Methods One hundred and fourteen newborns with NRDS were randomly divided into treatment group(58 cases) and control group(56 cases),on the base of same combined therapy,the cases in control group only underwent oxygen-absorbing by head set with the flow rate 4-6 L/min,and the cases in treatment group were given large dose mucosolvan(Ambroxol Hydrochloride) 30 mg/(kg?d) + 5%GS 20 mL,for two times and variant flow rate continuous positive airway by nasal mask(NCPAP),the parameter setting flow rate 6-8 L/min,FiO_2 0.4-0.6,pressure 5-8 cm H_2O.The clinical symptom and blood gas analysis after 12 and 48 hours were observed and compared the changes of pa(O_2),pa(CO_2),pa(O_2)/FiO_2 in two groups.Results The dyspnea and groan in 44 cases in the treatment group lessoned or vanished,pa(O_2) rised and pa(CO_2) lowered,the oxygenation index obviously increased,the cases with RDS grade Ⅰand gradeⅡ had better therapeutic effect,and the cases with RDS grade Ⅲ(X-ray)and Ⅳ had not manifest effect,the total effective rate was 75.8% in treatment group and 26.7% in control group.There were significant difference in therapeutic effect and oxygenation index between two groups.Conclusions Large dose mucosolvan(combi)-ning variant flow rate continuous positive airway by nasal mask can significantly improve the ventilation and oxygenation function and there are significant therapeutic effect in NRDS,especially in the NRDS grade Ⅰand gradeⅡ,the trachea cannula may be avoided and mechanical ventilation rate may be decreased if the therapeutic method can be used in earlier period.