1.Effects of miR-135a on HOXA10 expression, proliferation and apoptosis of ovarian cancer cells
Weiwei TANG ; Guiping WAN ; Yicong WAN ; Lin ZHANG ; Wenjun CHENG
Chinese Journal of Obstetrics and Gynecology 2013;(5):364-369
Objective To investigate the effects of miR-135a on HOXA10 expression,proliferation and apoptosis of SKOV3 cells.Methods (1) Through computer-aided algorithms,the predicted target gene of miR-135a (HOXA10)were determined.(2) miR-135a mimics,miR-135a inhibitor and negative control were transfected into SKOV3 cells,respectively.Reverse transcription (RT)-PCR,western blot analysis were used to examine the expression levels of HOXA10 at different times (24,48 and 72 hours).(3) A luciferase reporter assay was used to confirm the direct regulation between miR-135a and HOXA10.(4) SKOV3 cells proliferation at different times (24,48 and 72 hours) was detected by methyl thiazolyl tetrazolium(MTT) assay [quantified by absorbance(A)].Western blot was used to examine the expression of apoptosis-associated protein bcl-2,bax and caspase-3 in SKOV3 cells after 48 hours transfection.Results (1) HOXA10 was predicted to be the target gene of miR-135a by computer-aided algorithms.(2) RT-PCR shown that HOXA10 mRNA levels were decreased over time (24,48 and 72 hours) after miR-135a mimics transfectionin SKOV3 cells (0.94 ±0.04 vs 0.78 ±0.03 vs 0.70 ±0.03,P <0.05).While,the expression of HOXA10 mRNA was increased over time after miR-135a inhibitor transfection (1.14 ± 0.05 vs 1.16 ±0.03 vs 2.60 ±0.08,P <0.05).After transfected with miR-135a mimics or miR-135a inhibitor over 48 and 72 hours,the HOXA10 expression levels in SKOV3 cells were significantly lower or higher than each control group,respectively (all P < 0.01).Western blot analysis of HOXA10 expression in SKOV3 cells confirmed the results of RT-PCR detected.(3) After cotransfection of miR-135a plasmid and pMIR-REPORT luciferase plasmid containing HOXA10,luciferase reporter assays showed that the luciferase activity reduced by 67.8% (P <0.01).(4) MTT showed that SKOV3 cells growth after miR-135a mimics transfection for 48 and 72 hours were significantly lower than those in control group (0.38 ± 0.03 vs 0.52 ± 0.05,0.67 ±0.05 vs 0.75 ± 0.06 ; respectively,all P < 0.05).While,SKOV3 cells transfected with miR-135a inhibitor for 72 hours grew significantly faster than that in control group (0.95 ± 0.05 vs 0.75 ± 0.06,P < 0.01).After miR-135a mimics transfection,the level of bcl-2 protein was significantly lower than that in control group (0.28 ±0.06 vs 0.76 ±0.09,P <0.01).The activity of caspase-3 was significantly higher than that in control group (115.0 ± 2.4 vs 95.4 ± 2.1,P < 0.01).While,there was no statistical difference of bax expression (P =0.142).However,after miR-135a inhibitor transfection,the expression level of bcl-2 protein was significantly higher than that in control group (0.92 ± 0.03 vs 0.76 ± 0.09,P =0.037) and the activity of caspase-3 was significantly lower than that in control group (59.5 ± 4.1 vs 95.4 ± 2.1,P < 0.01).There was also no statistical difference of bax expression (P =0.066).Conclusion miR-135a may play an important role in cell proliferation and apoptosis of ovarian cancer cells by regulating HOXA10 and its downstream pathways.
2.Evaluation of the clinical screening and the follow-up for early colorectal cancer by colonoscopy in the elderly
Jun WAN ; Ziqi ZHANG ; Cheng ZHU
Chinese Journal of Geriatrics 2001;0(05):-
Objective To evaluate the importance of clinical screening and follow up by direct colonoscopy for colorectal cancer at an early and curable stage. Methods There were 2 196 elderly people aged between 60 to 89 years. The clinical screening by direct colonoscopy was performed according to the protocol. 1 740 of 2 196(79.2%) patients were followed up every year. Results Fifty two elderly persons were found to be colorectal cancer patients by colonoscopy, with the detectable rate being 2.4%. Nineteen were diagnosed early stage colorectal cancer, accounting for 36.5% of the detected colorectal cancer. Nine among the followed up cases were detected early colorectal cancer, accounting for 45 0% of the detected colorectal cancer. The resectable rate and the 5 year survival rate was 97 7% and 80 9% for colorectal cancer, respectively. 98 9% of the cecum intubation cases was successful. The incidence of complication for colonoscopy was 0 05%. Conclusions By clincal colonscopy screening and follow up study for colorectal cancer and precancerous changes in the elderly, the patients with adenomatoid polyps were early diagnosed and treated, so it raised the detectable rate of early colorectal cancer and the level of grade prevention of colorectal cancer.
3.The Educational Objectives-Oriented Requirements of Medical Students' Core Competence
Cheng ZENG ; Wei ZHANG ; Xiaoyan WAN
Chinese Journal of Medical Education Research 2002;0(01):-
Based on B.S. Bloom's taxonomy of educational objectives, development of medical students' core competence requirements is paid great attention to meet the needs of society within worldwide medical education community. The Institute for International Medical Education has developed the “Global Minimum Essential Requirements in Medical Education” of medical students' core competence oriented in the educational objectives of physicians.
6.Application of ISSR-PCR to identification of different Dendrobium Sw. species
Ying SHEN ; Cheng XU ; Xiaofeng WAN ; Ming ZHANG
Chinese Traditional and Herbal Drugs 1994;0(03):-
Objective To identify nine species of Dendrobium Sw. by inter-simple sequence repeats-polymerase chain reaction (ISSR-PCR) method and to investigate the difference among the species of Dendrobium Sw. at DNA molecular level. Methods Ten primers constituted by simple sequence repeats (SSR) were tested for PCR and sepharose electrophoresis of the nine species. Results Seven of the ten primers amplified polymorphic bands. The number of polymorphic band positions of each primer ranged from 7 to 14, and the size of fragments ranged from 220 to 1 260 bp. The amplification patterns of two primers, namely UBC-807 and UBC-864, were higher in terms of polymorphic and amplified band ratio. Each of them was able to distinguish all the examined species. Conclusion ISSR-PCR provides a quick, reliable molecular marker technique for identification of different species of Dendrobium Sw.
7.Long-term dynamic observation of erythromycin eye ointment entered into the orbit of guinea pigs by using magnetic resonance imaging
Lu YANG ; Jingliang CHENG ; Yong ZHANG ; Jingjing WAN ; Yu ZHU
Journal of Practical Radiology 2014;(6):1030-1034
Objective To evaluate the MRI features of erythromycin eye ointment entered into the orbital soft tissue of guinea pigs.Methods MRI was performed dynamically to observe the signal changes 1 day,1 week,2 weeks ,3 weeks and 4 weeks after erythromycin eye ointment was successfully injected into orbital soft tissue of guinea pigs ,as well as HE stainings to monitor retrob-ulbar soft tissue specimens at each time point were obtained.Results After the injection of erythromycin eye ointment 1 day,1 week,2 weeks,3 weeks and 4 weeks later,the signal intensity of erythromycin eye ointment was resemble to the surrounding soft tissue and indistinguishable on T2 WI.On T1 WI,the signal intensity of erythromycin eye ointment was higher than that of contralat-eral adipose body of orbit at 1day and 1 week,and slightly higher than the contralateral adipose body of orbit at 2 weeks and 3 weeks,but resemblance at 4 weeks.HE staining showed that inflammatory cell infiltration and tissue necrosis at 1 day,the presence of foreign body macrophages and fibroblasts at 2 weeks,and the inflammatory cells surrounding the optic nerve gradually subsided at 3 weeks and 4 weeks.Conclusion Fat-suppressed T1 WI is the best sequence to observe erythromycin eye ointment.Abnormal signal changes on T1 WI can reflect the histopathological changes after erythromycin eye ointment into the tissue.
8.A hemodynamic and hemolysis study of the axial pump with hydrodynamic-magnetically levitated impeller in swine
Lufeng ZHANG ; Zhiming SONG ; Xinghua CHENG ; Guorong LI ; Feng WAN
Chinese Journal of Thoracic and Cardiovascular Surgery 2015;31(4):239-241
Objective To explore the in-vivo hemodynamic and hemolysis effect of a newly designed axial continuousflow ventricular assist device(VAD) in swine.Methods Under general anesthesia,each of 5 swine [weight (40.0 ± 5.2)kg] was implanted with the axial continuous-flow VAD into the apex of left heart ventricle,and the outflow graft was anastomosised to descending aorta.Results All of the axial continuous-flow VAD were implanted successfully with post-operative survival rate 100%.All 5 animals survived over one week.There was a positive correlation between pump speed and assistance effect.The mean left ventricular systolic pressure was (131.6 ± 28.0) mmHg(1 mmHg =0.133 kPa).While the axial continuous-flow VAD was working,left ventricular end diastolic pressure decreased,along with mean intraventricular pressure declined.Peripheral hemodynamics was stable and peripheral blood pressure was not remarkably different from the pressure preoperation.Daily urine volume was in normal range within 1 week post operation.Free hemoglobin in plasma was slightly elevated on the surgery day,and gradually dropped to normal level within 1 week.International Normalized Ratio(INR) was maintained between 2.0-2.5 with oral adminiatration of warfarin of 3 mg/day.There was no thrombosis existing in VAD at autopsy.Conclusion The application of the axial pump with hydrodynamic-magnetically levitated impeller in animal experiment can provide stable hemodynamics,advanced heart unloaded effect,favorable peripheral perfusion,and blood compatibility is satisfactory.
9.Purification and Characterization of L-AI from Bacillus Stearothermophilis IAM 11001 Expressed in E.coli
Li-Fang CHENG ; Wan-Meng MU ; Tao ZHANG ; Bo JIANG ;
China Biotechnology 2006;0(09):-
Thermostable L-arabinose isomerase (L-AI) is the most potential enzyme for the biological production of D-tagatose from D-galactose, a novel functional factor. Gene araA encoding the L-arabinose isomerase from Bacillus stearothermophilis IAM 11001 was cloned and expressed in Escherichia coli. The araA gene of 1491 bp has 95% identity with L-AI from Thermus sp. IM6501. The GenBank accession number for the nucleotide sequence of this araA gene determined in this work is EU394214.The bacterium was induced by IPTG and analyzed by SDS-PAGE, approximately 59 kDa exogenous protein was observed on the SDS-PAGE. The recombinant L-AI was purified to electrophoretical homogeneity with affinity chromatography, and the activity of recombinant L-AI was also studied. The bioconversion rate of D-galactose to D-tagatose reached 39.4% after 24h whole cell reaction.
10.Research Progress on the Microbial L-Arabinose Isomerase
Li-Fang CHENG ; Wan-Meng MU ; Tao ZHANG ; Bo JIANG ;
Microbiology 2008;0(10):-
L-arabinose isomerase (L-AI) can isomerize L-arabinose and D-galactose into L-ribulose and D-tagatose, respectively, which is currently the most effective biological catalyst for D-tagatose production. The crystal structure of L-AI has been solved recently and its gene has been cloned, sequenced and overex- pressed. L-AI improved by protein engineering will be the dominant enzyme for industrial production of D-tagatose. This paper reviewed researches on protein structure and function, properties and application in D-tagatose production of L-AI, and the long-term potential development of L-AI was prospected.