Humans ; Inflammation ; Lipoproteins, HDL ; blood ; chemistry

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.

OBJECTIVETo evaluate the efficacy and safety of Jinying Capsule (JC) in treating pelvic inflammatory disease patients with accumulated damp-heat syndrome (ADHS).

METHODSTotally 328 patients were recruited in a prospective, positive drug parallel controlled, and multi-center clinical trial. Of them 213 patients in the treatment group took JC (0.5 g per capsule), 4 capsules each time, 3 times per day, while 115 patients in the control group took Kangfuyan Capsule (KC, 0.4 g per capsule), 3 capsules each time, twice per day. The course of treatment was 4 weeks for all. Scores of Chinese medical syndromes, visual analogue scale (VAS) of the lower abdominal pain, and European quality of life-five dimension scale (EQ-5D) were observed before treatment and after 4 weeks of treatment.

RESULTSThere were 204 patients in the treatment group and 109 in the control group who completed this trial. The total effective rate of Chinese medical syndrome was 89.71% (183/204 cases) in the treatment group and 76.15% (83/109 cases) in the control group (P < 0.01). Compared with before treatment in the same group, EQ-5D scores increased, and VAS scores of the lower abdominal pain decreased in the two groups after treatment. EQ-5D scores was 0.857 ± 0.157 in the treatment group, obviously higher than that in the control group (0.753 ± 0.126, P < 0.05). VAS scores of the lower abdominal pain was 2.14 ± 1.23 in the treatment group, lower than that in the control group (2.33 ± 1.24), but with no statistical difference between the two groups (P > 0.05). No adverse reaction occurred in the two groups.

CONCLUSIONJC was superior to KC in improving Chinese medical syndrome and quality of life of pelvic inflammatory disease patients with accumulated damp-heat syndrome.

Capsules ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Hot Temperature ; Humans ; Medicine, Chinese Traditional ; Pelvic Inflammatory Disease ; drug therapy ; Phytotherapy ; Prospective Studies ; Quality of Life ; Safety ; Syndrome

OBJECTIVETo explore the function of vascular endothelial cell (VEC) in patients with coronary heart disease (CHD) of Xin-blood-stasis syndrome.

METHODSSome vasoactive substances produced by VEC were detected and analyzed in patients with CHD of or without Xin blood stasis syndrome in group A (n=112) and group B (n=108) respectively, also in patients with non-CHD but of Xin-blood-stasis syndrome in group C (n=110), and healthy persons in group D (n=100), including nitric oxide (NO), endothelin (ET), angiotensin H (Ag II), soluble intercellular adhesion molecule-1 (sICAM-1) and soluble vascular cell adhesion molecule -1 (sVCAM-1).

RESULTSThe abnormality degree of ET, Ag II , sICAM-1 and sVCAM-1 in various groups showed such a tendency as group A> group B> group D (P < 0.01 or P < 0.05), while no significant difference in these criteria between group A and group C was shown (P > 0.05).

CONCLUSIONThe vasoactive substances secreted by VEC are closely related to the formation and progression of CHD, and are likely to be important pathological markers of blood-stasis syndrome in CHD.

Adult ; Aged ; Aged, 80 and over ; Coronary Artery Disease ; physiopathology ; Diagnosis, Differential ; Endothelial Cells ; metabolism ; physiology ; Endothelins ; metabolism ; Female ; Humans ; Intercellular Adhesion Molecule-1 ; metabolism ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Nitric Oxide ; metabolism

According to the designed specific primers of gene fragment based on the Salvia miltiorrhiza transcriptome data, a full-length cDNA sequence of SQS2 from S. miltiorrhiza f. alba was cloned by the method of reverse transcription polymerase chain reaction (RT-PCR). The SmSQS2 cDNA sequence was obtained, this sequence is named SmSQS2 and its GenBank registration number is KM244731. The full length of SmSQS2 cDNA was 1245 bp, encoding 414 amino acids including 5'UTR 115 bp and 3'UTR 237 bp. Sequence alignment and phylogenetic analysis demonstrated that SmSQS2 had relative close relationship to the SQS2 of S. miltiorrhiza. The induction of E. coli [pET28-SQS2] in different temperature, induction time, IPTG concentrations and density of inducing host bacterium (A600) were performed, Shaking the culture at 30 degrees C until the A600 is approximately 0.6 and add IPTG to final concentration of 0.2 mmol x L(-1), and then the optimal expression of SmSQS2 recombinant protein were accumulated after the induction time of 20 h. The research provided important base for the study of sterol and terpene biosynthesis of SQS2 in S. miltiorrhiza f. alba.
Cloning, Molecular ; Farnesyl-Diphosphate Farnesyltransferase ; chemistry ; genetics ; metabolism ; Models, Molecular ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism ; Salvia miltiorrhiza ; chemistry ; classification ; enzymology ; genetics ; Sequence Alignment

OBJECTIVETo investigate therapeutic effects of vacuum sealing drainage (VSD) in the treatment of soft tissue defect combined with tendon and bone exposure.

METHODSFrom October 2007 to February 2011, 397 patients (412 feet) with open ankle fracture and dislocation combined with soft tissue defected were treated by VSD. There were 301 males and 96 females with an average age of 36 years (ranging age from 20 to 73 years). According to AO classification, 74 feet were type I, 211 feet were type II, 108 feet were type III and 19 feet were type IV. The mean time from injury to operation was 5.6 h ( 2 to 12 h). The mean treatment time of was 10 months (4 to 19 months).

RESULTSOne hundred and forty-one patients were primarily healed, 97 patients were sutured at stage II. Split-thick skin grafting was performed at stage II was performed in 103 patients; free flap transplantation was performed in 25 patients. Three of the 34 patients with infection were removed steel plate; Eviscerate flap coverage wound was performed in 14 patients caused by the first metatarsal bone exposure; Toe amputation were performed in 22 cases caused by toes necrosis. Tarsometatarasl joints perforators' surgery was performed in 10 patients with forefeet necrosis. Thirty hundred and six patients were followed up from 3 to 20 months (averaged 10 months). The wounds healed well.

CONCLUSIONVSD for soft tissue defects caused by ankle injury is a simple and effective method, but can not replace debridement and transfer flap.

Adult ; Aged ; Ankle Fractures ; Debridement ; Drainage ; methods ; Female ; Humans ; Joint Dislocations ; surgery ; Male ; Middle Aged ; Skin Transplantation ; Treatment Outcome ; Vacuum ; Young Adult

Objective To understand the prevalence,drug resistance and clinical features of children with acute lower respiratory tract infection(ALRI) caused by escherichia coli.Methods From Oct. 2005 to Oct. 2006,659 patients with ALRI who were admitted to hospital were chosen and their nasopharyngeal secretions were obtained and cultured.K-B disc diffusion for antibiotic susceptibility were performed for these clinical isolates.Results Among 659 patients,118 cases were isolated escherichia coli,the rate was 17.99% which had 90 boys and 28 girls.Eighty-seven of 118 E.coli strains were with extended-spectrum ?-lactamase(ESBLs),the rate was 73.73%.All of strains were sensitive to imipenem.For ESBLs-producing strains,the ratio of resistance tocefotaxime,ceftriaxone,cefuroxime,ampicillin,piperacillin were 78.81%,73.73%,73.73%,76.27%,78.81%,respectively.Conclusions The positive rate of ESBLs producing E coli in Kunming area is high and drug resistance is severe gradually.Imipenem can be the first selection for treatment on these infections.

Acyl carrier protein is an essential component involved in the biosynthesis of DHA(Docosahexaenoic Acid) via PKS(Polyketide synthase) pathway,which takes the growing acyl chain from one enzyme to another.One cDNA clone,with high homology of ACP,was isolated from Schizochytrium sp.FJU-512 cDNA library.The deduced amino acid sequence contained 142 residues with isoelectric point of 5.04 and had the 4'-phosphopantetheine prosthetic(4'-PP) binding site.The target fragment was digested with BamHⅠ/HindⅢand inserted into the expression vector pET-30a resulting in the plasmid pET-30a/acp.The recombinant vector was transformed into E.coli BL21(DE3) and induced by IPTG.SDS-PAGE analysis demonstrated that ACP was effectively expressed.

Objective:To construct a DNA vaccine co-expressing the HBV compound multi-epitope antigen gene, the hIL-12 and the anti-HBV siRNA genes, and to express this DNA vaccine in HepG2 cells. Methods:The HBV multi-epitope antigen gene was designed and synthesized before it was fused with enhanced green fluorescent protein(EGFP) gene, and cloned into the multi-clone site(MCS) of the eukaryotic expression vector pVAX1. The expressinig units of hIL-12 and siRNA were cloned into the BspH I and Mlu I site of pVAX1 respectively. Then the recombinant plasmid pVAX1-siHBV-HB-EGFP-hIL12 was transiently transfected HepG2 cells. The expression of HBV compound multi-epitope gene was observed through EGFP report gene. The expression of hIL-12 was analyzed by ELISA and the effects of anti-HBV siRNA was confirmed with rtPCR . Results: The analysis of enzyme digestion and sequencing both demonstrated that the trible-expressing HBV DNA vaccine has been constructed successfully. The green fluorescent image was detected in the transfected cells which could confirm the expression of the multi-epitope antigen gene. The amount of hIL-12 secretion was 1289pg/ml in supernatant at 48h after transfection and 1712pg/ml at 72h after transfection. The mRNA amount of HBV S gene, which was the siRNA target, had been obviously knockdown. Conclusion: The DNA vaccine co-expressing the HBV compound multi-epitope antigen gene, the hIL-12 and the siRNA genes was constructed and transiently expressed in HepG2 cells, and siRNA had shown us a good anti-HBV effect. It laid a foundation of further study on anti-HBV effect of the new DNA vaccine.

20(Z= -2.117, P=0.034), and between the patients with OD and without OD (Z=-3.089, P=0.002), but PCT was not so between the non-surviror and survivor (Z=-1.307, P=0.191). The serum PCT level correlated with the incidence of organ dysfunction (x~2=14.82, P=0.033) and APACHEII (x~2=12.83, P