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.
3.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.
4.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.
5.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.Does urothelial cancer of bladder behave differently in young patients?
Zhi-Hua WANG ; You-Yuan LI ; Zhi-Quan HU ; Hui ZHU ; Qian-Yuan ZHUANG ; Yong QI ; Zhang-Qun YE
Chinese Medical Journal 2012;125(15):2643-2648
BACKGROUNDBladder urothelial cancer has been diagnosed at an increasing rate among young adults in China while the clinical outcomes remain highly controversial. To optimize the management of young patients with bladder cancer, we examined whether bladder urothelial cancer in young patients behaved differently from that in the elder patients.
METHODSFrom 1994 to 2008, a database of bladder urothelial cancer patients at a major tertiary medical center was retrospectively reviewed. The clinical and pathological parameters of patients who were less than 40 years of age and a series of patients older than 40 years of age as the control group during the same period were compared. A survival analysis was performed using the Kaplan-Meier method and log-rank test, and Cox regression was performed to identify clinical parameters that affected the clinic outcomes.
RESULTSYoung bladder cancer patients had a lower male-to-female ratio and were less likely to have advanced stages and high-grade cancers at the initial diagnosis. Tumors in young bladder cancer patients tended to be less multifocal at diagnosis. In addition, young patients had a lower recurrence rate and longer recurrence interval than older patients. The Kaplan-Meier curve and Log-rank test showed that young patients had significantly better cancer specific survival than old patients. The univariate and multivariate Cox regression analysis revealed that tumor grade is the sole predictor for tumor recurrence in young patients.
CONCLUSIONSYoung patients with bladder cancer have favorable pathological features and clinical outcomes than older patients. These findings argue for more conservative management approaches for young patients with bladder cancer.
Adolescent ; Adult ; Age Factors ; Child ; Female ; Humans ; Male ; Retrospective Studies ; Sex Factors ; Urinary Bladder Neoplasms ; pathology ; Urothelium ; pathology ; Young Adult
9.Combined use of thin-section CT andF-FDG PET/CT for characterization of solitary pulmonary nodules.
Yun-Yan REN ; You-Cai LI ; Hu-Bing WU ; Quan-Shi WANG ; Yan-Jiang HAN ; Wen-Lan ZHOU ; Hong-Sheng LI ; Zhen WANG ;
Journal of Southern Medical University 2017;37(3):283-289
OBJECTIVETo investigate whether fluorine-18 fluorodeoxyglucose positron emission tomography/computed tomography (F-FDG PET/CT) combined with thin-section CT improves the diagnostic performance for solitary pulmonary nodules (SPNs).
METHODSA total of 267 patients underwent examinations withF-FDG PET/CT and thin-section CT for evaluating the SPNs with undetermined nature, which was further confirmed by pathological examination or clinical follow-up. The performance of two diagnostic criteria based on findings in PET/CT alone (Criterion 1) and in PET/CT combined with thin-section CT (Criterion 2) were compared.
RESULTSThin-section CT provided greater diagnostic information for SPNs in 84.2% of the patients. Compared with Criterion 1, the diagnosis based on Criterion 2 significantly increased the diagnostic sensitivity (80.4% vs 91%, P<0.01) and accuracy (76.4% vs 87.2%, P<0.01) for lung cancer. The lesion size and the CT features including lobulation, air bronchogram, and feeding vessel, but not SUVmax, were all helpful for characterizing non-solid SPNs. Thin-section CT rectified diagnostic errors in 50% (20/40) of the cancerous lesions, which had been diagnosed as benign by PET due to their low metabolism. For non-solid SPNs, Criterion 2 showed a significantly higher diagnostic sensitivity than Criterion 1 (90.0% vs 40.0%, P=0.000) but their diagnostic specificity were comparable (75.2% vs 58.3%, P=0.667). For solid nodules, the use of thin-section CT resulted in no significant improvement in the diagnostic performance (P<0.05).
CONCLUSIONThe combination of PET/CT and thin-section CT creates a synergistic effect for the characterization of SPNs, especially non-solid nodules.
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