1.Construction and identification of plasmid mediated RNA interference targeting Nogo-66 receptor
Journal of Third Military Medical University 2003;0(13):-
Objective To construct and identify of a plasmid mediated high efficiency of RNA interference(RNAi) against Nogo-66 receptor(NgR).Methods After cloning of NgR by RT-PCR,the fragments were inserted into pcDNA3.1/CT-GFP-TOPO to produce plasmid expressing NgR-GFP fusion protein.Four pairs of oligonucleotide were designed according to NgR sequence and annealed.The resulting fragments were ligated into short hairpin RNA(shRNA) expressing plasmid.The plasmids expressing NgR-GFP fusion protein and shRNA were co-transfected into AAV-293 cells.RNAi efficiency against NgR was observed under fluorescent microscope and calculated by Western blotting.Results The interference efficiency of one sequence was above 90%.Conclusion A plasmid mediated high RNAi efficiency against NgR is constructed successfully.
2.Screening and identification of differentially expressed genes in hippocampus of mice during hypobaric hypoxic delayed preconditioning
Youming FAN ; Yuqi GAO ; Jian HUANG ; Weigong LIAO ; Mingchun CAI
Chinese Journal of Pathophysiology 1986;0(02):-
AIM: This study was designed to explore the differentially expressed genes between hypobaric hypoxic delayed preconditioning (HHDP) and normal mouse hippocampus. METHODS: HHDP was produced by treating the animals at a 7 000 m high altitude for 2.5 h/d for 3 d. At 36 h after last time decompression, total RNA was isolated from hippocampus. cDNA was synthesized and amplified by SMART PCR. cDNA libraries of differentially expressed gene between HHDP and control hippocampus were constructed. 452 clones from forward (subtracted control from preconditioning) cDNA library and 74 clones from backward (subtracted preconditioning from control) one were screened by reverse Northern hybridization. RESULTS: Screening with subtracted probes, hybridization signal of 85 gene fractions decreased and that of 217 gene fractions increased by more than 2 times in HHDP hippocampus compared with control. Screening with unsubtracted probe, hybridization signal of 44 gene fractions decreased and that of 135 gene fractions increased by more than 2 times in HHDP hippocampus compared with control. Some of the clones had been sequenced. Analysis and comparison with the data of GenBank were performed. The results showed that mouse cytochrome C oxidase subunit 1, NADH dehydrogenase subunit 1 and 6, deleted in split-hand/split-foot 1 region (DSS1) and cDNA corresponding to clone IMAGE: 5251089 of mice cDNA library were increased in hippocampus of HHDP mice. cDNA corresponding to clone IMAGE: 3593193, mus musculus adult male olfactory brain cDNA and mus musculus bladder RCB-0544 MBT-2 cDAN were decreased in hippocampus of HHDP mice. CONCLUSION: Many genes expresses differentially in hippocampus of mice during HHDP. This may be one of the molecular mechanisms of HHDP.
3.Effects of acute and chronic hypoxia on rat brain mitochondrial translation activity
Mingchun CAI ; Junze LIU ; Liping WU ; Bingyon SUN
Chinese Journal of Pathophysiology 1999;0(09):-
AIM: To investigate the effect of simulated high altitude hypoxia on rat brain mitochondrial translation activity. METHODS: Animals were continuously exposed to simulated 4 000 m high altitude in hypobaric chamber for three days and forty days. Mitochondria of rat brain were isolated by homogenizing brain tissue and following centrifuging program. Protein translation activity in isolated mitochondria in vitro was measured with -Lencine incorporation method. Products labeled with -methionine in isolated mitochondrial protein synthesis system in vitro were separated on SDS-PAGE and identified by autoradiography. RESULTS: Mitochondrial translation activity in vitro in acute hypoxia exposure were significantly lower than control( P
4.Effect of hyperin on acute liver injury in rats against oxidative stress-induced by CCl4
Mingchun HUANG ; Jianhong CHEN ; Xiaogang HU ; Peiyuan XIA ; Yongqing CAI ; Qiang WANG
Journal of Regional Anatomy and Operative Surgery 2013;(6):588-590,593
Objective To study the protective effect of hyperin( Hyp) on the acute liver injury in rats induced by CCl4 . Methods The acute liver injury model was induced by CCl4 . The effect of Hyp on acute liver injury of rat liver histopathology was observed;and the impact of changes of homogenates total superoxide dismutase ( T-SOD) ,glutathione ( GSH) activity and malondialdehyde ( MDA) in liver were de-tected. Results The acute liver injury of tissue induced by CCl4,apparent pathological inflammation and fibrous tissue degeneration and necrosis were observed by HE staining;At the high doses of 60 mg/kg and medium doses of 30 mg/kg treated by Hyp,liver pathology changes was significantly obvious. The T-SOD,GSH activity of liver tissue was significantly increased in the groups of Hyp treatment,MDA content was significantly decreased,which related to the dosages. Conclusion In the rat model of acute liver injury induced by CCl4 . A better therapeu-tic effect of Hyp was shown,the mechanism may be related to its antioxidant activity.
5.Course construction and practice of progress in pathophysiology for postgraduate students
Junze LIU ; Qingyuan HUANG ; Gang ZHANG ; Wenxiang GAO ; Xiaoling TAN ; Weigong LIAO ; Youming FAN ; Mingchun CAI ; Jian CHEN
Chinese Journal of Medical Education Research 2012;11(1):25-28
Proceeding from the requirements of teaching target for postgraduate students,the course of progress in pathophysiology was constructed and administrated.The course objective was defined which combined teaching knowledge with fostering students' ability together,especially the ability to think new ideas and to do scientific research.Aiming at this teaching target,the teaching contents which combined with the direction of scientific research of the department was growing together with scientific development,especially in new knowledge and new technique.Multiple teaching means and several mode of examine were adopted during the process of teaching practice.