1.Construction and regulated expression of mouse TGF-?1 recombinant adenovirus vector
Yan ZHANG ; Yulian XU ; Qian SHEN
Chinese Journal of Pathophysiology 1989;0(06):-
AIM: To construct a recombinant adenovirus vector which regulated expression of mouse transforming growth factor beta (TGF-?1). METHODS: Total RNA was extracted from Balb/c mice spleen pre-treated with Con A to clone TGF-?1. pTRE-shuttle vector was used as mediator to ligate TGF-?1 gene and the backbone of the replication-incompetent adenoviral vector. The constructed recombinant adenovirus contains tetracycline-responsive element which can regulate the expression of inserted genes. Identifying the desired recombinant adenovirus, it was packaged in HEK 293 cells. Supernatant of high titer adenovirus was collected to detect the TGF-?1 gene expression by ELISA kit. RESULTS: The constructed recombinant adenovirus had been identified by restriction endonucleases cutting, sequencing and PCR. In the supernatant of infected cells, it expressed the mice TGF-?1 in high concentration to 91.16 ng/L. CONCLUSION: The mice TGF-?1 recombinant adenovirus has been built which make a basis for further research and use in gene therapy.
2.Effects of probucol on the NF-κB-like activity of aortic smooth muscle cell in vitro
Jiangtao GU ; Zonggui WU ; Qian SHEN ; Yulian XU
Academic Journal of Second Military Medical University 2001;22(2):151-154
Objective:To observe the inhibitory effects of pr obucol on nuclear factor-κB (NF-κB)-like activity of vascular smooth mus cle cells(VSMCs). Methods:The effects of probucol and H2O2 on NF-κB-like activity of VSMCs were investigated by electrophoretic mobility shift assay(EMSA). Results:NF-κB activation of VSMCs could be induced by H2O2 or new born calf serum(NCS) for 72 h, the gray-measure of the protein bindings measured through computer scanning was 63.9 and 46.6 respe c tively. NF-κB activity of VSMCs stimulated by H2O2 or NCS might be prevent ed by 100 μmol/L probucol, the inhibitory rate was 37.1%, 14.8% respectively. Conclusion:probucol can inhibit NF-κB activity of VSMCs stim ulated by NCS or H2O2. These effects of probucol might help to re duce development of atheroclerosis and restenosis after percutaneous transluminal coronary angioplasty.
3.Effect research of aliskiren on bradykinin and high sensitive C-reactive protein in hypertension patients
Fengyi GUO ; Zhenhong HUO ; Yulian SHEN ; Dayan ZHANG
Chinese Journal of Postgraduates of Medicine 2012;35(30):23-25
Objective To explore the value of a novel non-peptide renin inhibitor aliskiren on hypertension patients,and analyze the change of bradykinin (BK) and high sensitive C-reactive protein (hs-CRP).MethodsEighty patients with mild to moderate hypertension were selected.Patients were randomized divided into ramipril group,aliskiren 75 mg/d group,aliskiren 150 mg/d group,aliskiren 300 mg/d group with 20 cases each by random digits table method and received the following types of intervention:ramipril 5 mg/d,aliskiren 75 mg/d,aliskiren 150 mg/d and aliskiren 300 mg/d.Enzyme-linked immunosorbent assay method was used for detecting hs-CRP.Radioimmunoassay method was used for detecting BK.Results After treatment,the levels of BK and hs-CRP in aliskiren 75 mg/d group,aliskiren 150 mg/d group and aliskiren 300 mg/d group [(5.06 ± 1.61),(5.05 ± 1.87),(5.27 ± 1.39) μg/L and (0.38 ± 0.11 ),(0.25 ± 0.05),(0.33 ± 0.11 ) mg/L] were significantly lower than those in ramipril group [ (7.12 ± 1.12) μ g/L,(0.49 ± 0.19) mg/L ] (P<0.05 ).After treatment,the levels of SBP and DBP had no significant difference among the four groups (P>0.05).Conclusion Aliskiren has a very good effect in decreasing blood pressure and does not cause the levels of BK and hs-CRP increasing.
4.The effect of inhalation budesonide in the treatment of cough variant asthma and its influence on serum cytokine
Zhenhong HUO ; Fengyi GUO ; Yulian SHEN ; Dayan ZHANG
Chinese Journal of Primary Medicine and Pharmacy 2012;19(17):2583-2584
Objective To observe the effect of inhalation budesonide(pulmicort) in the treatment of cough variant asthma and(CVA) its influence on serum cytokine.Methods 112 children with CVA were randomly divided into the atomization group and control group,each group 56 cases.The atomization group was given pulmicort inhalation therapy,and the control group was given oral long-acting β2 agonist treatment.After 3 months of treatment,the efficacy and serum cytokine levels were compared between the two groups.Results 3 months after the treatment,the effective rate of the atomization group was 91.07%,and significantly higher than that of the control group(75.00% )( x2 = 5.13,P < 0.05 ) ; The serum interleukin-4,γ-interferon,IgE,eosinophil counts of the atomized group were significantly lower than those of control group ( t =2.10,2.34,27.63,3.13,all P < 0.05 ).Conclusion The effect of inhalation pulmicort in the treatment of cough variant asthma is good,and has good safety.
5.Effects of probucol on the NF-?B-like activity of aortic smooth muscle cell in vitro
Jiangtao GU ; Zonggui WU ; Qian SHEN ; Yulian XU ;
Academic Journal of Second Military Medical University 1982;0(02):-
Objective:To observe the inhibitory effects of probucol on nuclear factor ?B (NF ?B) like activity of vascular smooth muscle cells(VSMCs). Methods:The effects of probucol and H 2O 2 on NF ?B like activity of VSMCs were investigated by electrophoretic mobility shift assay(EMSA). Results:NF ?B activation of VSMCs could be induced by H 2O 2 or new born calf serum(NCS) for 72 h, the gray measure of the protein bindings measured through computer scanning was 63.9 and 46 6 respectively. NF ?B activity of VSMCs stimulated by H 2O 2 or NCS might be prevented by 100 ?mol/L probucol, the inhibitory rate was 37.1%, 14.8% respectively. Conclusion:probucol can inhibit NF ?B activity of VSMCs stimulated by NCS or H 2O 2. These effects of probucol might help to reduce development of atheroclerosis and restenosis after percutaneous transluminal coronary angioplasty. [
6.Effect of Treadmill Exercise on Expression of Matrix Metalloproteinase-2 in Rats with Traumatic Brain Injury
Xiafeng SHEN ; Shan TIAN ; Yi WU ; Yulian ZHU
Chinese Journal of Rehabilitation Theory and Practice 2017;23(5):525-528
Objective To determine the effect of treadmill exercise on expression of matrix metalloproteinase-2 (MMP-2) in rats follow-ing traumatic brain injury (TBI). Methods Fifty-five male adult Sprague-Dawley rats were randomly assigned to sham (n=15), control (n=20) and exercise (n=20) groups, the later two groups subjected to unilateral cortical contusion injury (CCI). All the rats were assessed with foot-fault test 6, 12, 18 and 24 days after CCI. Evans blue perfusion was used to evaluate the integrity of the blood-brain-barrier (BBB) 48 hours after CCI. Protein expression of MMP-2 was determined with Western blotting one week after CCI. Results The score of foot-fault test improved more in the exercise group than in the control group 12, 18 and 24 days after CCI (F>4.793, P<0.05). Evans blue extravasa-tion was less in the exercise group than in the control group (t=-8.091, P<0.001), as well as the expression of MMP-2 (t=-13.12, P<0.001). Conclusion Early treadmill exercise can improve the motor function in rats with TBI, which may associate with inhibition of MMP-2 ex-pression to protect BBB integrity.
7.Pathological observation of airway inflammation after neonatal CVB3 inoculation in rats
Chaoping FANG ; Feng FANG ; Qian SHEN ; Yulian XU ; Dalie MA ; Hongxia WEI
Academic Journal of Second Military Medical University 2001;22(5):472-474
Objective: To observe the effect of coxsackie virus B3 on airway tract and lung morphology, and to study the relation between CVB infection and asthma. Methods: We established CVB3 infective model: 5 d neonatal rats inhaled CVB3 by ultrasonic brume. CVB3-IgM was examined 10 d after inoculating of CVB3, and LW/BW, airway tract and lung pathological change 10 d and 30 d after inoculation of CVB3 were observed. Results: Rats from the virus group had higher D of CVB3-IgM than control's (+2s ) and had higher LW/BW 10 d after inoculation of CVB3 than control (P<0.01). Neonatal rats had acute inflammatory changes 10 d after inoculation of CVB3 and persistent changes in morphology and cytology. Conclusion: Neonatal rats virus model is established. Respiratory infection by CVB3 in neonatal rats has persistent changes in airway tract inflammatory and morphology.
8.The Clinical Investigation of Mycophenolate Mofetil for the Prevention of Acute Rejection
Yulian JI ; Yanqiang YANG ; Guangbao YANG ; Xueqing YU ; Zongpei JIANG ; Qingrui SHEN ; Peigen WU ; Lanying ZHU
Journal of Sun Yat-sen University(Medical Sciences) 2001;22(3):215-217
【Objective】To investigate the role of mycophenolate mofetil (MMF) in the prevention of acute rejection in renal transplantation.【Methods】A total of 106 patients were randomized into two groups.One group received MMF (n=56),the other received azathioprine (Aza) (n=50).The time of the following study was within the first 6 months after transplantation.【Results】The rate of acute rejection of group receiving MMF was 20%,it′s lower than that of the group receiving Aza 44% (P<0.01).The recovery rate of acute rejection treated by methylprednisolone (MP),in MMF group 82% was higher than Aza group 55%.(P<0.05).Meanwhile the hepatotoxicity as well as cytomegalovirus (CMV) infection were lower in MMF group than those in Aza group.【Conclusion】MMF as a new anti-rejection drug could more effectively prevent acute rejection than Aza after renal transplantation,and has lower toxicity and side effect.
9.Pathological observation of airway inflammation after neonatal CVB_3 inoculation in rats
Chaoping FANG ; Feng FANG ; Qian SHEN ; Dalie MA ; Hongxia WEI ; Yulian XU ;
Academic Journal of Second Military Medical University 1985;0(05):-
Objective: To observe the effect of coxsackie virus B 3 on airway tract and lung morphology, and to study the relation between CVB infection and asthma. Methods: We established CVB 3 infective model: 5 d neonatal rats inhaled CVB 3 by ultrasonic brume. CVB 3 IgM was examined 10 d after inoculating of CVB 3, and LW/BW, airway tract and lung pathological change 10 d and 30 d after inoculation of CVB 3 were observed. Results: Rats from the virus group had higher D of CVB 3 IgM than control's ( +2s ) and had higher LW/BW 10 d after inoculation of CVB 3 than control ( P
10.Expression of exon 13 from the Ki-67 gene in human cells and tissues by digoxigenin-labelled mRNA in situ hybridization
Yulian WU ; Chenghong PENG ; Hongwei SHEN
Chinese Medical Journal 2001;114(1):48-53
Objective To get insight on the regulatory mechanism of Ki-67 gene expression in malignant cell cycle. Methods Non-radioactive in situ hybridization (ISH) was undertaken, combined with immunohistochemistry to study the Ki-67 gene transcription and translation in various human cells and tissues. HeLa cells and fresh colon cancer cells, tonsil, normal pancreas and pancreatic cancer tissues were used in this study. A 435?bp cDNA fragment located in exon 13 of the Ki-67 antigen gene was amplified by polymerase chain reaction (PCR). Digoxigenin-labelled antisense and sense RNA probes were prepared for detecting Ki-67 mRNA, combined with MIB-1 immuno~histochemistry. Results Successful localization of Ki-67 mRNA in human HeLa cells, colon cancer cells, tissues specimen of the tonsil and pancreatic cancer tissue sections was accomplished by digoxigenin-labelling in situ hybridization technique. ISH to colon cancer cells and pancreatic cancer tissue slides showed that much stronger cytoplasm and perinuclear mRNA signals of the Ki-67 gene were present in malignant cells than in normal cells, which was in accordance with MIB-1 nuclear protein signals. Conclusions A sensitive and practical in situ hybridization method for the analysis of Ki-67 antigen mRNA in human cell and tissue was developed. Abnormal transcription of exon 13 of Ki-67 gene might be responsible for malignant cell proliferation in colon and pancreatic cancer.