2.Construction of serine protease gene mutant and study on pathogenicity of serine protease gene of Entero-coccus faecalis
Li-Xian WU ; Wen HUANG ; Sun XIANG ; Ping XIAO ;
Chinese Journal of Infectious Diseases 2000;0(02):-
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.
3.Evaluation on Efficacy and Safety of Jinying Capsule in Treatment of Pelvic Inflammatory Disease Patients with Accumulated Damp-heat Syndrome.
Qin LI ; Chun-yan CHEN ; Yu-ping SUO ; Min HUANG ; Xian-hua HUANG
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(12):1459-1462
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
4.Selection of bFGF Mimic Peptide by Phage Display
Hui-Xian HUANG ; Xiao-Ping WU ; Shao-Hui CAI ; Xiao-Kun LI ;
China Biotechnology 2006;0(05):-
Objective: To obtain the bFGF mimic peptide binding to FGFR via phage display, and to provide the base for developing peptide agonist of bFGF. Methods: Using Balb/c 3T3 cells as the target cells and COS-7 cells as the subtractive panning, the phage display heptapeptide library was biopanned for 4 rounds to obtain the single phage clones. The affinity and the specificity of the clones were assessed by ELISA. DNA sequencing was applied to further analyze the positive clones. Results: Twelve positive clones were selected from the enriched phages. A group of hydrophobic peptides containing a conserved motif, PR, was identified. Conclusion: Two bFGF mimic heptapeptides binding to FGFR were selected, which may be used as the candidates for bFGF agonist.
5.Cloning and Expression of Acyl Carrier Protein Gene from Schizochytrium
Zhi-Ping LI ; Xian-Zhang JIANG ; Bao-Yu TIAN ; Zheng-Yu SHU ; Jian-Zhong HUANG ;
Microbiology 1992;0(02):-
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.
6.Study on a Novel Hepatitis B Virus Multi-epitope DNA Vaccine with siRNA,hIL-12 Coexpressing
Jing-Xian HUANG ; Yi-Cheng CAO ; Zheng-Ping DU ; Chang-Li TAO ; Hua-Qiang YANG ;
China Biotechnology 2006;0(08):-
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.
7.Pterygium conjunctive reverse transplantation combined with amniotic membrane transplantation on recurrent pterygium
Ting, LI ; Shu-Xian, CHEN ; Xu-Guang, XIA ; Zheng, YIN ; Mu-Zhen, HUANG ; Ping-Ying, GUO
International Eye Science 2014;(9):1715-1716
To discuss the effective method of decreasing the postoperative recurrence rate of recurrent pterygium.
●METHODS:Totally 126 cases (126 eyes) with recurrent pterygium were randomly divided into A group (56 cases) and B group ( 70 cases ). Group A was treated by pterygium conjunctive reverse transplantation combined with amniotic membrane transplantation, group B was treated by amniotic membrane transplantation. The followed-up time after surgery was 6-24mo.
●RESULTS:ln group A, postoperative 5-7d (average 5. 62± 1. 38d), cornea epithelium was repaired. ln group B, postoperative 7- 10d ( average 7. 38 ± 1. 12d), the corneal wound was healed. There was statistical significant difference between two groups (t = 4. 307,P<0. 05). Three cases recurrence were noted in A therapeutic group (56 cases), the recurrent rate was 5. 4%; Twelve cases recurrence were noted in B compared group (70 cases), the recurrent rate was 17. 1%. There was statistical significant difference between two groups(P<0. 05).
●CONCLUSlON: lt is suggested that pterygium conjunctive reverse transplantation combined with amniotic membrane transplantation is effective in the treatment of recurrent pterygium.
8.Clinical Analysis on 118 Children with Acute Lower Respiratory Tract Infection Caused by Escherichia Coli
ming-bo, ZHAO ; qian, WU ; yong-kun, HUANG ; lin-xian, NI ; ping, LU ; jing, XU
Journal of Applied Clinical Pediatrics 2003;0(10):-
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.
9.The relationship with fat liver in children with simple obesity
Dong-Ping LIU ; Xian-Ming GONG ; Chun-Hua LI ; Hai-Zhong HUANG ;
Chinese Journal of Primary Medicine and Pharmacy 2006;0(07):-
Objective To search the relationship among blood pressure,blood lipid and fat liver in childrcn with simple obesity.Methods Blood pressure,blood lipid and ultrasound of liver of 40 cases with simple obesity and 20 cases in normal control had been detected.Results there were 11 cases without complication in children with simple obesity,the simple obesity with hypertension complication were 14 cases,and the simple obesity with high blood lipid were 15 cases.The level of total cholestrrol(TC),triglyceride(TG),light density lipoprotein(LDL)and very light density lipoprotein(VLDL)in obesity children were significantly higher than those of control group(P
10.Relationship of serum levels of PCT and organ disfunction in patients with severe multiple trauma
Jun FEI ; Hong-Jun YU ; Hua-Ping HANG ; Xian-Kai HUANG ; Yao-Guang JIANG ;
Chinese Journal of Emergency Medicine 2006;0(06):-
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