2.Cloning and Expression of dauW Gene from Daunorubicin-producing Streptomyces coeruleorubidus SIPI-1482
He HU ; Ke SHANG ; You-Jia HU ; Bao-Quan ZHU ; Chun-Bao ZHU ;
China Biotechnology 2006;0(12):-
A novel gene,located between dnrX and drrB in the genome of daunorubicin-producing strain Streptomyces coeruleorubidus SIPI-1482,was cloned and named as dauW.The full sequence of dauW was submitted to GenBank(Accession No.EF523565).Blast result indicated that it showed high homology with dnrW in GenBank.The exact function of dauW is as yet unknown despite the possibility that it might belong to a family of FAD-dependent oxidoreductases on the basis of conserved domain analysis.dauW was cloned into expression plasmids pET-28a(+)and pET-32a(+),respectively,and was successfully expressed in E.coli DE3 after induction with IPTG.The preliminary results of the expression of dauW suggested that it might be involved in the self resistance in Streptomyces coeruleorubidus due to the increased resistance to daunorubicin in the E.coli host.
4.Disruption and Compensation of dnmV Gene from Daunorubicin-producing Strain Streptomyces coeruleorubidus SIPI-1482
Ke SHANG ; Qian GONG ; You-Jia HU ; Chun-Bao ZHU ; Bao-Quan ZHU ;
China Biotechnology 2006;0(07):-
TDP-4-ketohexulose reductase, encoded by dnmV, is important in daunorubicin biosynthesis. To obtain a daunorubicin block mutant, double cross-over plasmid pYG817 was constructed by inserting apramycin resistant gene and amplified dnmV together with upstream dnmU into vector pUC18. dnmV was successfully disrupted after transformation of daunorubicin-producing strain SIPI-1482 by pYG817. Daunorubicin was absent from metabolites of the resulting transformant, and its biosynthesis can be reconstituted by introducing dnmV expression plasmid into the disruptant, although the yield is lower than wild-type SIPI-1482, according to HPLC analysis. This mutant can be a good candidate for production of anthracycline such as epi-daunorubicin by introducing exogenous gene into the host.
5.DNA Shuffling of Vitreoscilla Hemoglobin
Ning YUAN ; You-Jia HU ; Chun-Bao ZHU ; Bao-Quan ZHU ;
China Biotechnology 2006;0(11):-
To improve the growth enhancement activity of Vitreoscilla hemoglobin(VHb), Vitreoscilla hemoglobin gene(vgb) was mutated by error-prone PCR and then reconstituted by DNA shuffling. The shuffling library was constructed by inserting the shuffled genes into the downstream of vgb natural promoter and transforming them into E.coli DH5?. Mutated active VHb proteins were first screened in test tubes according to host cell pellets color and then in shake flasks according to host pellets wet weight .One active mutant protein, VHb′042506, was obtained after second screening. It could increased the host wet weight by 31.25% and 58.75% than that of the control which bearing natural VHb under microaerobic and extremely microaerobic conditions, respectively. Sequencing and alignment results showed that 11 nucleotides were mutated, thus resulted in 4 amino acids changes occurred in this mutant protein. CO difference spectrum test also indicated that it had higher specific absorption.
6.Expression and Characterization of Thrombin Inhibitor TTI
Li-Ping XIE ; Ke SHANG ; Chun-Bao ZHU ; Bao-Quan ZHU ; You-Jia HU ;
China Biotechnology 2006;0(10):-
TTI gene coding for Tsetse thrombin inhibitor was modified with E.coli bias codon and expressed in Escherichia coli with high efficiency.Recombinant protein was purified to more than 98% purity.Assay for enzyme activity determination was set up.The result showed that the fusion protein exhibited inhibiting activity for thrombin.Inhibitory rate of purified TTI was 73% when concentration of thrombin and substrate was 10U/ml and 250?mol/L respectively.Inhibition pattern was determined as competitive with Ki at 35?mol/L.
7.Tandem Repeat Expression and Inhibitory Activity of a ?-lactamase Inhibitory Peptide
Ming-Fei XU ; Li-Ping XIE ; Tao YANG ; Chun-Bao ZHU ; Bao-Quan ZHU ; You-Jia HU ;
China Biotechnology 2006;0(09):-
?-lactamase inhibitor research is popular for its potential on ?-lactam antibiotics resistant strain.A ?-lactamase binding peptide SIPIS04-01 was obtained by the yeast two-hybid system.In vitro assay showed that it can inhibit the ?-lactamase activity.In order to improve the expression level of the recombinant peptide,a two-copy expression plasmid pYG563 was constructed by random orientation tandem repeat method after codon modification,the two-copy plasmid was successfully expressed and the product was increased by 48.4% than that of one-copy plasmid.Purified peptide showed inhibitory activity against TEM-1 ?-lactamase in vitro and the inhhibitory constant Ki was measured.
8.A case-control study on correlation of pesticide exposure with childhood acute leukemia.
Yan ZHANG ; Sha ZHU ; Yu GAO ; Xiao-jin WANG ; Tao CHEN ; You YANG ; Guo-quan WANG ; Guo-hua HU ; Rong SHI ; Ping JIN ; Ying TIAN
Chinese Journal of Preventive Medicine 2011;45(1):41-46
OBJECTIVETo evaluate the correlation of pesticide exposure with childhood acute leukemia.
METHODSAn exploratory case-control study was conducted among childhood acute leukemia patients under 15 years old in Shanghai, China. From January 1st, 2006 to December 31st, 2008, a total of 80 newly diagnosed acute leukemia patients were recruited from Shanghai Children's Medical Center for the case group. Another 96 age-matched patients who visited the hospital for health examination, pediatric treatment or osteological therapy excluding hematological system diseases and neoplastic disease, were recruited for the control group. A questionnaire survey was conducted in both groups; and a 30 - 40 ml random urine sample was collected from each participant. Five types of organophosphorus pesticide metabolites was then detected among the samples, using Gas Chromatography with Flame Spectrophotometry.
RESULTSAccording to result of the questionnaire survey, more participants (55.0% (44/80)) in case group than in the control group (33.3% (32/96)) reported using mosquitocidal, which might increase the risk of childhood acute leukemia (OR = 2.444; 95%CI: 1.326 - 4.506). At the same time, the detection showed that the concentration (median) of organophosphate metabolites diethyl phosphate, dimethyl phosphate, dimethyl thiophosphate, diethyl thiophosphate and diethyl dithiophosphate in case group were 0.0682, 0.0082, 0.0183, 0.0233, 0.4259 µg/g Cr, which were all significantly higher than in control group (0.0865, 0.0025, 0.0112, 0.0123, 0.1207 µg/g Cr) except the concentration of diethyl phosphate (Z = -1.081, P = 0.279). The difference showed statistical significance (Z = -5.752, -2.800, -3.316, -8.120, P < 0.05).
CONCLUSIONPesticide exposure may be one of the risk factors for childhood acute leukemia.
Acute Disease ; Adolescent ; Case-Control Studies ; Child ; Child, Preschool ; Environmental Exposure ; adverse effects ; Female ; Humans ; Infant ; Infant, Newborn ; Leukemia ; etiology ; Male ; Maternal Exposure ; Pesticides ; adverse effects ; Pregnancy ; Risk Factors
9.Association of haplotype formed on HLA-DRB1 and HLA-DQA1 alleles with outcomes of hepatitis B virus infection.
Liang-ping LU ; Xing-wang LI ; Ying LIU ; Guo-chang SUN ; Zhi-hai CHEN ; Xi-lin ZHU ; Quan-you HU ; Hui LI
Chinese Journal of Medical Genetics 2006;23(4):427-430
OBJECTIVETo assess the association of haplotype of HLA-DRB1 and HLA-DQA1 alleles with outcomes of hepatitis B virus infection in Han population of north China.
METHODSTwo hundred and seven chronic hepatitis B (HB) patients, two hundred and twelve chronic asymptomatic hepatitis B virus (HBV) carriers (HBV carrier) and one hundred and forty-eight self-limited HBV infection were investigated for HLA-DRB1 and HLA-DQA1 alleles by sequence specific-polymerase chain reaction (PCR-SSP).
RESULTSThe frequency of DRB1*04-DQA1*0301 haplotype was 10.03% in self-limited HBV infection subjects, significantly higher than that in chronic HB patients (3.66%) (P=0.0005)ûthe frequency of DRB1*15/*16-DQA1*0102 haplotype was 6.80% in self-limited HBV infection subjects, significantly higher than 1.94% in chronic HB patients (P=0.0012) and 1.65% in asymptomatic HBV carriers (P=0.0004)ûwhile the frequency of DRB1*04-DQA1*0302 haplotype was 3.10% in chronic HB patients, higher than that in self-limited HBV infection subjects (0.39%) (P=0.0077).
CONCLUSIONIndividuals with different haplotypes composed of HLA-DRB1 and HLA-DQA1 might have different outcomes of HBV infection.
Adolescent ; Adult ; Alleles ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; genetics ; HLA-DQ Antigens ; genetics ; HLA-DQ alpha-Chains ; HLA-DR Antigens ; genetics ; HLA-DRB1 Chains ; Haplotypes ; Hepatitis B ; genetics ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction
10.Chuzhen therapy for sub-health: a randomized controlled study.
You-Ping HU ; Zhen-Jie XU ; Jia WU ; Lin XING ; Lei-Xiao ZHANG ; Jin-Xing ZHANG ; Hui TAN ; Shi-Zhi MAO ; Cong-Quan ZHOU
Chinese Acupuncture & Moxibustion 2012;32(11):1019-1023
OBJECTIVETo observe the effect on intervention of sub-health with pestle needle (Chuzhen).
METHODRandomized controlled trail was adopted for this research. One hundred and fifty-three cases were randomly divided into two groups of a Chuzhen group (79 cases) and a massage group (74 cases). Acupoint of Bazhen (Baihui Bazhen, Shendao Bazhen, Zhiyang Bazhen, Mingmen Bazhen, Yaoyangguan Bazhen), Hechelu on the head, the neck and the lumbar area were adopted in Chuzhen group. While regular whole-body massage was applied in the massage group. The human sub-health score, the cornell medical index (CMI) and thermal texture maps system (TTM) technology of the two groups before and after the intervention were observed.
RESULTS1) After treatment, sub-health condition score, the CMI score, the M-R score and the TTM index were all increased in both groups (all P<0.01) 2) Comparison of D-value of the two groups before and after the intervention: the level of the sub-health score, the total score of CMI, and the index of sleep, pressure, Governor Vessel, Hukou (first web), blood lipid, viscosity of blood, microcirculation of TTM index of the Chuzhen group changed more obvious (all P<0.01), but there was no statistic significances in the M-R score and blood sugar of the TTM (both P>0.05). 3) The sub-health condition score in Chuzhen group was higher than that in the massage group (P<0.01).
CONCLUSIONChuzhen therapy has definite effect on intervention of sub-health, which is better than regular general massage.
Acupuncture Points ; Acupuncture Therapy ; Adolescent ; Adult ; Aged ; Cornell Medical Index ; Female ; Humans ; Male ; Massage ; Middle Aged ; Young Adult