3.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.
4.Disrupting sfa1 Gene to Enhance Biosynthesis of Ethanol in Saccharomyces cerevisiae
Hao-Lei SONG ; Xiao-Xian GUO ; Yan-Zun WANG ; Xian-Zhang JIANG ; Jian-Zhong HUANG ;
Microbiology 1992;0(03):-
The sfa1 gene encoded a bifunctional enzyme with the activities of both alcohol dehydrogenase and glutathione-dependent formaldehyde dehydrogenase in Saccharomyces cerevisiae.The gene disruption cassette produced by PCR using the same long oligonucleotides which comprise 19 or 22 nucleotides complementary to sequences in the templates(pUG6 and pUG66 marker plasmid)at 3' end and 45 nucleotides at 5' end that annealed to sites upstream or downstream of the genomic target sequence to be deleted.After two linear disruption cassettes with a Cre/loxP mediated marker were transformed into the cells of Saccharomyces cerevisiae YS-1,the positive transformants were checked by PCR to correct the integration of the cassette and concurrent deletion of the chromosomal target sequence.Once correctly integrated into the genome,the select marker can be efficiently rescued by transformating the plasmid pSH47 into YS-1 and inducing the Cre expression with a Cre/loxP-mediated marker removal procedure.The expression of the Cre recombinase finally resulted in the removal of the marker gene,leaving behind a single loxP site at the chromosomal locus.The diploid mutant YS-1-sfa1 was generated,which could enhance the output of ethanol with 8.0% by shaking culture in flask compared with the original strain YS-1.
5.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.
6.Perioperative fluid management during pediatric kidney transplantation
Peijiao LIAO ; Cuiling SU ; Yun XIAO ; Xiaoling LIANG ; Lijuan HUANG ; Xian REN
Modern Clinical Nursing 2013;(8):35-37
Objective To explore the strategies and effectiveness of perioperative fluid administration during pediatric kidney transplantation.Methods In our study,we evaluated the heart function and compliance of pediatric patients preoperatively,and then formulated perioperative fluid management strategy according to children’s circulation feature.The electrolytic balance was kept and complications were prevented.Results Twenty-nine out of all 48 children experienced electrolyte disturbance to different extents on day 1 postoperatively,which improved on day 2 after prompt management.There were five cases of oliguria,three cases of delayed graft function(DGF),one case of heart failure and multiple organ failure respectively.Conclusion Such nursing strategies like formulation of reasonable fluid infusion plan according to the physiological features of pediatric recipients,keeping the electrolytic balance and careful management of fluid at polyuria,oliguria and anuria,care critical for the children to live through the perioperative periods.
7.Combination of interferon α with mannan peptide in treatment of HBeAg-positive chronic hepatitis B
Xiao LING ; Zhenxiang TANG ; Shuquan CHENG ; Yongchao XIAN ; Xin YE ; Yifeng CAI ; Chengjun HUANG ; Hui NI
Chinese Journal of Clinical Infectious Diseases 2009;2(5):268-272
Objective To investigate the clinical effect of IFNα combined with mannan peptide in treatment of patients with HBeAg-positive chronic hepatitis B ( CHB ). Methods Eighty HBeAg-positive CHB patients with HBV DNA quantity ranging from 10 to 10 eopies/mL were enrolled and randomized into the treatment group and the control group ( n = 40 for each ). Patients in treatment group were given daily subcutaneous injection of IFNα-2b 5,000,000 U for 52 weeks, and received mannan peptide 10 mg per intravenous injection or 2. 5 mg per intramuscular injection for a total of 2 to 3 treatment courses (12 weeks for each). The control group received only IFNα-2b treatment. Liver function, serum markers of hepatitis B, HBV DNA quantity and blood tests were performed before the treatment and at 2, 4, 8, 16, 26 and 52-week during the treatment; and the adverse effects were recorded. Results The rates for ALT normalization, negative HBsAg, negative HBeAg, HBeAg seroconversion and negative HBV DNA were 91. 8% , 17. 5% , 52. 5% , 27. 5 % and 47. 5% at 52nd week in the treatment group, while those in the control group were 80. 0% , 12. 5% , 30. 0% , 10. 0 % and 25. 0% , respectively. There were significant differences in HBeAg-negative, HBeAg-seroeonversion and HBV DNA-negative rates between two groups (χ2 = 4. 178, 4.021 and 4.381, P < 0. 05 ) , and these indexes in the treatment group were increased to 57. 5% , 30. 0% and 50. 0 respectively at 52nd week after drug withdraw. White blood cells began to be elevated at 4th week and were restored to the normal levels at 8th week in the treatment group, while the count in the control was lower than the normal value even at 52nd week of the treatment with the average of (3.45±1. 18)×109/L. Conclusion Alpha-interferon combined with mannan peptide therapy is effective for patients with HBeAg-positive CHB, which may restore the declined peripheral WBC counts induced by interferon and improve the compliance.
9.Analysis on principle of treatment of phlegm retention syndrome of Ma Peizhi based on apriori and clustering algorithm.
Jia-Rui WU ; Wei-Xian GUO ; Xiao-Meng ZHANG ; Xiu-Qin HUANG ; Bing YANG
China Journal of Chinese Materia Medica 2014;39(4):631-633
Analyzed the prescriptions for phlegm retention syndrome that built by Ma Peizhi by the association rules and clustering algorithm, the frequency of drug usage and the relationship between drugs could be get. And from that we could conclude the experiences for phlegm retention syndrome of Ma Peizhi of menghe medical genre. The results of the analysis were that 18 core combinations were dig out, such as Citri Exocarpium Rubrum-Eriobotryae Folium-Citri Reticulatae Pericarpium. And there were 9 new prescriptions were found out such as Aurantii Fructus-Citri Exocarpium Rubium-Eriobotryae Folium-Citri Reticulatae Pericarpium. The results of the analysis were proved that Ma Peizhi of Menghe Medical Genre was good at curing phlegm retention syndrome by using the traditional Chinese medicine of mild and light, such as the medicines of mild tonification, and clearing damp and promoting diuresis.
Algorithms
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Data Mining
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Databases, Factual
;
Drug Prescriptions
;
standards
;
Drugs, Chinese Herbal
;
therapeutic use
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Humans
;
Mucus
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chemistry
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Practice Patterns, Physicians'
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Respiratory Tract Diseases
;
drug therapy
10.Analysis on medication rule in prescriptions for hemoptysis of Menghe physician Ma Peizhi based on knowledge-discovery in database.
Jia-Rui WU ; Wei-Xian GUO ; Xiu-Qin HUANG ; Bing ZHANG ; Xiao-Meng ZHANG
China Journal of Chinese Materia Medica 2014;39(4):627-630
The clinical prescriptions of hemoptysis that built by Menghe physician Ma Peizhi were collected, and were analyzed by the method of unsupervised data mining, such as association rules and clustering algorithm. From the prescriptions, we got the frequency of drugs, the association rules among drugs, 10 core drug combinations and 5 new prescriptions. Accordingly, after the analysis, Menghe physician Ma Peizhi had rich experience in the treatment of hemoptysis. His therapy for hemoptysis was clearing heat and moistening lung, dispel heat from blood to stop bleeding, and enriching yin and blood. And we can also make a conclusion that the TCM inheritance support system is of great practical value for minning the old doctors' clinical experiences.
Data Mining
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Databases, Factual
;
Drug Prescriptions
;
standards
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Drugs, Chinese Herbal
;
chemistry
;
therapeutic use
;
Hemoptysis
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drug therapy
;
Humans
;
Practice Patterns, Physicians'