1.Effects of inactivation of vitamin D receptor in intestinal tumor growth of APCmin/+ mice
Hong KANG ; Wei ZHENG ; Hong YU
Journal of Chinese Physician 2009;11(9):1174-1177
Objective To explore effects and mechanism of inactivation of vitamin D receptor (VDR) in intestinal tumor growth of APCmin/+ mice. Methods To generate APCmin/+VDR -/- mice through breeding, the number and size of small intestinal and colonic tumors were assessed and compared between APCmin/+ and APCmin/+ VDR -/- mice. The intestinal tumors were diagnosed with HE stain. The expressions of BCL-2,vimentin-1,Stat-1 and MSH-2 proteins of tumors were determined by immanohistocbemistry and compared be-tween APCmin/+ and APCmin/+VDR -/- mice. Results A comparison between APCmin/+ and APCmin/+ VDR -/- mice revealed that the number of the tumors, which were larger than 3mm, was significantly increased in APCmin/+ VDR -/- mice at 4 months of age (P< 0.01). HE staining indicated fistulous adenomas from small intestine and colon of two groups, Immunostaining showed Stat - 1 level in APC-min/+ tumors were increased and MSH-2 and vimentin-1 levels in APCmin/+ VDR-/- mice were increased, compared to APCmin/+tumors. Conclusion These observations suggested that inactivation of VDR promoted the intestinal tumor growth of APCmin/+ mice.
2.Determination of 10-Hydroxy-2-decenoic acid in Chongcaojing Wangjiang Capsule by HPLC
Jianpin YU ; Jie FENG ; Wei ZHENG
Traditional Chinese Drug Research & Clinical Pharmacology 2000;0(06):-
To determine the content of 10-Hydroxy -2-decenoic acid in Chongcaojing Wangjiang Capsule by HPLC.The column was Discovery C 18 .The detective wavelength was at 209 nm. The mobile phase was acetonitrile: 0.5% phosphoric acid (25∶75) with the flow rate being 1.0 mL/min and the column temperature at 25℃.There was a good linearity (r=1) winthin the range of 0.02144 ~ 0.38592 ?g. The average recovery rate was 101.8%, RSD= 1.6%. [Conclusion] This method can be used for the quality control standard of Chongcaojing Wangjiang Capsule.
3.Effects of ELOVL4 gene overexpress on the synthesis efficiency of n3 and n6 very long chain polyunsaturated fatty acids
Man, YU ; Wei, LIN ; Bo, CHEN ; Zheng-Zheng, WU
International Eye Science 2014;(8):1386-1390
AIM:To compare the synthesis efficiency of n3 and n6 very long chain polyunsaturated fatty acid ( VLC-PUFA ) by overexpressing ELOVL4 protein, providing guidance for treating Stargardt-like macular dystrophy (STGD3).
METHODS:To establish recombinant adenovirus with the ELOVL4 protein and green fluorescent protein, transferred into cultured PC12 cells. The cells were divided into 3 groups: PC12, PC12 + Ad- GFP and PC12 + Ad-ELOVL4, former two groups serve as controls. ELOVL4 gene expression was quantified by qRT-PCRs. ELOVL4 protein was analyzed by Western - Blot ( WB ) . The transduced cells were treated with both EPA and AA (1:1). After 48h of incubation, cells were collected, total lipids extracted and fatty acid methyl esters prepared and analyzed by gas chromatography-mass spectrometry ( GC-MS) .
RESULTS:When supplemented together, 20:5n3 (EPA) and 20:4n6 ( AA) were efficiently taken up at almost the same amounts in the PC12 cells regardless of ELOVL4 expression. The ELOVL4-expressing cells elongated both EPA and AA to a series of n3 and n6 VLC-PUFAs. From 20:5n3/EPA, 34:5n3 and 36:5n3 account for 0. 71% and 1.6%, respectively. From 20:4n6/DHA, 34:4n6 and 36:4n6 were only 0. 46% and 0. 61%, respectively. The total relative mol% of n3 VLC-PUFAs synthesized from EPA was almost two times that of n6 VLC-PUFAs synthesized from AA.
CONCLUSION: ELOVL4 protein preferentially elongates n3 PUFA to VLC - PUFAs over n6 PUFA. Dietary supplementation of appropriate n3/n6 PUFAs may provide STGD3 patients with some therapeutic benefits.
4.Molecular typing of Salmonella paratyphi A strains isolated in Hangzhou area from 2002 to 2013
Haoqiu WANG ; Wei ZHENG ; Hua YU ; Wei ZHANG ; Jincao PAN
Chinese Journal of Microbiology and Immunology 2017;37(1):57-61
Objective To analyze the molecular characteristics of Salmonella paratyphi A ( S. pa-ratyphi A) strains prevailing in Hangzhou area in recent years. Methods Pulse field gel electrophoresis ( PFGE) and multiple-locus variable-number tandem repeat analysis ( MLVA) were performed for molecular typing and epidemiological analysis of 72 S. paratyphi A strains isolated in Hangzhou area during 2002 to 2013. Results The 72 S. paratyphi A strains were divided into 11 PFGE ( by using restriction enzymes of Xba Ⅰ and Bln Ⅰ) and 6 MLVA types. Among the selected 34 variable number tandem repeat ( VNTR) sites, 4 sites (1188K, 2075K, 2201K and 4346K) showed high polymorphism, in which PFGE displayed a higher resolution than MLVA. Except for the 5 PFGE types of X4B5, X7B7, X8B8, X9B9 and X10B10, the other 6 PFGE types belonged to a same clone sharing a similarity of greater than 95%, and the S. Paratyphi A strains in that clone accounted for 93. 1% of the total strains isolated in Hangzhou. Conclu-sion The occurrence of paratyphoid A in Hangzhou area from 2002 to 2013 was mainly caused by S. para-typhi A strains belonging to the same clone. Combination of PFGE with MLVA was conducive to epidemiolog-ical investigation of paratyphoid A.
5.Metabolic shift of Corynebacterium acetoacidophilum-deltaldh under oxygen deprivation conditions.
Qian YANG ; Pu ZHENG ; Fang YU ; Wei LIU ; Zhihao SUN
Chinese Journal of Biotechnology 2014;30(3):435-444
Lactate and succinate were produced by Corynebacterium acetoacidophilum from glucose under oxygen deprivation conditions. To construct knockout mutant, lactate dehydrogenase gene (ldh) of C. acetoacidophilum was deleted by double-crossover chromosome replacement with sacB gene. Comparing with the wild strain ATCC13870, ldhA-deficent mutant produced no lactate with glucose consumption rate decreased by 29.3%, while succinate and acetate concentrations were increased by 45.6% and 182%, respectively. Moreover, the NADH/NAD+ rate was less than 1 (about 0.7), and the activities of phosphoenolpyruvate carboxylase and acetate kinase of the ldhA-deficent mutant were enhanced by 84% and 12 times, respectively. Our studies show that succinicate and acetate production pathways are strengthened by blocking lactate synthesis. It also suggests that improving NADH supply and eliminating acetate generation are alternative strategies to get high succinate-producer.
Corynebacterium glutamicum
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genetics
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metabolism
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Glucose
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Industrial Microbiology
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L-Lactate Dehydrogenase
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genetics
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metabolism
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Lactic Acid
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metabolism
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Oxygen
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metabolism
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Phosphoenolpyruvate Carboxylase
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Succinic Acid
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metabolism
6.Expression of human papillomavirus L1 protein in lesions of condyloma acuminatum
Zhangyu BU ; Wei ZHENG ; Liming WU ; Hong ZHOU ; Xiaohong YU
Chinese Journal of Dermatology 2013;(2):130-132
Objective To investigate the expression of human papillomavirus (HPV) L1 protein in lesional tissue of condyloma acuminatum and its clinical significance.Methods Forty-three patients with condyloma acuminatum were included in this study,and received CO2 laser therapy.Tissue samples were resected from the lesions of these patients at their first visit before treatment.Immunohistochemistry was used to determine the expression of HPV L1 protein in these samples.Follow up was carried out once every 2 weeks for 3 months to assess the recurrence rate and frequency in these patients after treatment.Results HPV L1 protein was detected in 83.72% (36/43) of these tissue samples.The expression rate of HPV L1 protein decreased sequentially from patients infected with HPV 6/11,patients with both HPV 6/11 and 16/18,to those with HPV16/18 (x2 =17.90,P < 0.01).During the 12 weeks of follow up,the recurrence rate was 69.77% (30/43) with the average number of recurrence of 2.16.There was a sequential reduction in the recurrence rate and number from patients with strong expression of HPV L1 protein,to those with moderate expression,slight expression and negative expression (x2 =8.02,46.92,P < 0.05 and 0.01,respectively).The expression intensity of HPV L1 protein was negatively correlated with the recurrence rate and frequency (rs =-0.429,-0.696,respectively,both P < 0.01).Conclusion HPV L1 protein may serve as a helpful molecular biomarker for the prediction of prognosis and recurrence of condyloma acuminatum.
7.Induction chemotherapy combined with three-dimensional conformal radiation ther a py for locally advanced non small cell lung cancer
Aiqing ZHENG ; Jinming YU ; Xianguang ZHAO ; Xuetao WANG ; Guangsheng WEI
Chinese Journal of Radiation Oncology 1993;0(03):-
Objective To evaluate the effect and complication of inductio n chemot herapy combined with three-dimensional conformal radiation therapy (3DCRT) for l ocally advanced non small cell lung cancer (NSCLC). Methods Ninety-two such pa t ients were randomized into radiation therapy alone group(RT-, 50 patients) and i nduction chemotherapy combined radiotherapy group (CMT-, 42 patients). The indu c tion chemotherapy consisted of 2-4 cycles of platinum-based regimen. Results Th e overall median survival time was 15 months with 12 months in the RT group and 18 months in the CMT group(P=0.014)respectively. The 1-year o verall survival rates were 48.6% and 71.2% in RT and CMT group,respectively (P=0.004). The 2-year survival rates w ere 20.8% and 37.6% in RT and CMT group, respectively (P=0.0 41). Treatment was w ell tolerated and the toxicities were similar in either group. C onclusion The ad dition of induction chemotherapy to 3DCRT takes a survival advantage over 3DCRT alone for Stage Ⅲ NSCLC without increasing toxicities.
8.Expression and biological function identification of TCR?9/?2-Fc protein in baculovirus vector expression system
Yang GUO ; Jing ZHENG ; Yu HU ; Lianxian CUI ; Wei HE
Basic & Clinical Medicine 2006;0(12):-
Objective To establish an expression system of TCR?9/?2-Fc protein by baculovirus vector expression system and identify biological function of expressed TCR?9/?2-Fc protein.Methods ?9Fc and ?2(OT3)Fc gene fragments were amplified by overlap PCR and inserted into expression vector pBACp10ph.The recombinant plasmid pBACp10ph-?9/?2(OT3)-Fc and the baculovirus DNA were co-transfected into sf9 cells.The expression position of TCR?9/?2(OT3)-Fc was identified by Western blot and the expression efficiency of ?9Fc and ?2(OT3)Fc was tested by flow cytometry(FCM).Furthermore,the binding activity of TCR?9/?2(OT3)-Fc protein with SKOV3 cells and MNS protein was evaluated with laser scanning confocal microslopy and surface plasmon resonance(SPR).Results The recombinant vector pBACp10ph-?9/?2(OT3)-Fc was constructed and TCR?9/?2(OT3)-Fc protein was expressed in sf9 cells.However,the expression efficiency of ?9Fc and ?2(OT3)Fc was quite different.It was proved that purified TCR?9/?2(OT3)-Fc protein can bind with SKOV3 cell and MNS protein.Conclusion TCR?9/?2-Fc protein is successfully expressed in baculovirus vector expression system and TCR?9/?2-Fc protein can simulate the binding activity of TCR in vitro.
9.Duplication of animal models for spinal cord injury and its application in experimental therapeutics
Bing GU ; Zheng ZHANG ; Huanan LI ; Wei MENG ; Riyue YU
Chinese Pharmacological Bulletin 2009;25(12):1559-1562
The mechanism of spinal cord injury and repair therapy after nerve injury is currently a hotspot of neuroscience research.Duplicating animal models plays a key role in experimental therapeutics of spinal cord injury.This review systematically describes the progress in animal models for spinal cord injury including contusion, compression, transection, ischemic,distraction and chemical-mediated injury,which have been established at home and abroad.Based upon the aforementioned models,some applications in experimental therapeutics are simultaneously enumerated.All these information provides scientific guidance for the experimental novel drugs′screening.
10.Evaluation of multi-phase CT scanning for the liver transplantation
Hong ZHENG ; Xiaoping YU ; Wei CHEN ; Shuang YANG
Journal of Chinese Physician 2012;14(9):1193-1197
ObjectiveTo evaluation application of MSCTA technology in pre-and pro-liver transplantation.MethodsThirty - two patients with liver transplantation were performed with multi-phase CT scanning (MSCT).In hepatic artery and portal phase vascular 3D imaging reconstruction methods include multiple planar reconstruction ( MPR),maximum intensity projection ( MIP),volume rendering (VR).MIP images were measured with these parameters (CA,LGA,CHA,PHA,PV,SV,SMV in diameter).ResultsHepatic arterial vascular imaging scan can show clearly within the scope of the abdominal aorta,gastroduodenal artery,hepatic artery,the left hepatic,celiac trunk,and its branches.Portal vascular imaging clearly showed that the portal vein system.MIP can use accurate measurement of vascular diameter abdominal aorta and portal,superior mesenteric vein,and splenic vein diameter.Control group with liver cirrhosis and liver cancer artery diameter had no significant difference in patients with portal hypertension and the portal vein [ ( 16.7 ± 2.4 ) mm ],mesenteric vein [ ( 10.8 ± 2.1 ) mm ],and splenic vein diameter [(13.1±1.9)mm] compared to the control group[(13.8 ±1.6)mm,(8.2 ±1.7)mm,(8.9±1.1)mm ],the difference was statistically significant ( P < 0.05 ). Conclusions MSCTA can show the liver artery and portal vein system of the non-invasive method of checking,Joint MIP and VR application can provide more clinical liver transplant before the hepatic artery and portal vein of information,liver transplantation arteriovenous anastomoses provides for the monitoring of vascular diameter.