1.Correlation between focal adhesion kinase phosphorylation and intimal hyperplasia following vascular denudation .in rats
Cuiyu BAO ; Fangzhou CHENG ; Jun KE
Chinese Journal of Tissue Engineering Research 2007;11(49):10014-10016
BACKGROUND: Focal adhesion kinase (FAK) phosphorylation influences the proliferation, migration and apoptosis of cells. However, there are no reports about the role of FAK phosphorylation in migration and proliferation of vascular smooth muscle cells during the process of vascular restenosis.OBJECTIVE : To analyze the correlation between FAK phosphorylation and vascular remodeling after balloon endothelial denudation in rats.DESIGN: Random controlled experiments in animals.SETTING: Cardiology Institute of Xianning College; Shenzhen Center for Chronic Disease Prevention and Control;Department of Cardiology at the Affiliated Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology.MATERrALS: The experiment was carried out in the Laboratory of Cardiology at the Affiliated Tongji Hospital from March to May in 2005. Forty male Wistar rats were used in the study, and they were managed according to the ethical standard of animals.METHODS: ①Forty rats were randomized into five groups (n =8): non-balloon endothelial denudation control group, and 4-day, 8-day, 16-day and 24-day model groups after balloon endothelial denudation. ②The rat models of restenosis following balloon endothelial denudation were built. At days 4, 8, 16 and 24 postoperatively, the denudated vessels were harvested for the detection of the morphological changes, which were taken as the judgment of successful models;Western blot method was used to detect the expression and activity of FAK. Abdominal aorta of left common carotid in rats was served as controls.MAIN OUTCOME MEASURES: The expressions of FAK and phosphorylation FAK in rat artery.RESULTS: Totally 40 rats were all involved in the result analysis.①A small quantity of FAK without phosphorylation could be detected in the controlled rats. Compared with the control group, the expression and activity of FAK in the model group rapidly increased at 4 days after balloon injury (P < 0.05), and they changed in a similar manner with the increase of injury duration. The expression of FAK reached a peak at 8 days and began to descend at 16 days. The expression of phosphorylation FAK reached a peak at 16 days and then rapidly descended. ②Focal hyperplasia of intima was found at 8 days after balloon endothelial denudation, and diffuse hyperplasia of intima was found at 16-24 days after denudation.CONCLUSION: FAK can take part in vascular remodeling after balloon endothelial denudation in rats. The level of intimal hyperplasia is closely associated with the level of FAK and FAK phosphorylation.
2.Effectiveness of propafenone for paroxysmal supraventricular tachycardia
Jun KE ; Yuanzhong LI ; Fangzhou CHENG
Clinical Medicine of China 2001;0(07):-
Objective To observe the clinical efficacy and safety of propafenone on paroxysmal supraventricular tachycardia (PSVT).Methods 37 patients with PSVT were injected with 70 mg propafenone intravenously in 5 minutes.The unrecovered patients in 20 minutes were injected with 70 mg propafenone intravenously again,who were given 70 mg propafenone intravenously once again if not controlled in late 30 minuted.Blood pressure,heart beat,12 lead electrocardiogram were recorded before and after use of drug.Results The significant effective rate was 77.5% and effective rate was 21.6% with overall effective rate of 97.3% and effective time of 1~55 (7.1?2.8) minutes.The average accumulated dose of propafenone was 105 mg.Some side effects were observed in part of patients,including slight reduction of blood pressure,P R interval.QRS wave and Q T interval prolonged,temporary type sino atrial block,vertigo and nausea,etc whice could vanish gradually if not treated.Conclusion Propafenone by intravenous injection can quickly contro PSVT with saftey and effectiveness.
3.Interventional effect of telmisartan on expression and activation of focal adhesion kinase during vascular-injured remodeling in rats
Fangzhou CHENG ; Cuiyu BAO ; Ji PENG ; Jun KE
Chinese Journal of Tissue Engineering Research 2007;11(41):8401-8403
BACKGROUND:Adherent migration of vascular smooth muscle cells and proliferated remodeling of vessel walls following vascular endothelial injury play a key role in onset of restenosis after percutaneous transluminal coronary angioplasty,while expression and phosphorylation activation of focal adhesion kinase are attacked during this period.OBJECTIVE:To observe the interventional effect of angiotensin receptor antagonist,telmisartan,on the expression and activation of focal adhesion kinase during vascular-injured remodeling.DESIGN:Randomized controlled animal study.SETTING:Shenzhen Center for Chronic Disease Control and Prevention;Department of Cardiology,Affiliated Hospital of Xianning Medical College;Department of Cardiology,Tongji Hospital Affiliated to Tongji Medical College,Huazhong University of Science and Technology.MATERIALS:The experiment was carded out in the Cardiovascular Laboratory,Wuhan Tongji Hospital from March to May 2005.A total of 36 male Wistar rats weighing 300-360 g were randomly divided into control group,model group and treatment group with 12 in each group.METHODS:Aortal restenosis models were established after endothelial denudation.Foley's tube technique was used to strip vascular endothetium of rats in the model group and the treatment group;while,rats in the control group and the model group were fed and drunk normally; in addition, rats in the treatment group were also given 5 mg/(kg·d) telmisartan solution.Thirty clays after successive administration,vessels at injured sites were collected to observe the morphological changes and extract RNA and protein.Reverse transcriptase polymerase chain reaction (RT-PCR) was used to measure mRNA expression of focal adhesion kinase and Westem blot technique was USod to measure total protein and phosphorylated protein of focal adhesion kinase.MAIN OUTCOME MEASURES: Proliferation of vessel wall at injured sites; mRNA expression, total protein and phosphorylated protein of focal adhesion kinase at injured sites at 30 days after administration.RESULTS:A total of 36 rats Were involved in the final analysis.Thirty days after operation, aortic tunica intima was thickened in the model group, mRNA expression of focal adhesion kinase was increased, and total protein and phosphorylated protein were higher in the model group than those in the control group;however,proliferation of tunica 5ntima vasorum was lightened in the treatment group, activity of mRNA expression of focal adhesion kinase was decreased, and total protein and phosphorylated protein were lower in the treatment group than those in the model group.CONCLUSION: Telmisartan can remarkably relieve proliferation of tunica intima after denudation and inhibit expression and activation of focal adhesion kinase after vascular injury.Effect of telmisartan on vascular-injured remodeling may be related to inhibiting expression and activation of focal adhesion kinase.
4.Screening and Identification of The Proteins Interacting with The Transcription Factor X-box Binding Protein 1
Fengjin GUO ; Fangzhou SONG ; Faping YI ; Haien CHENG
Progress in Biochemistry and Biophysics 2006;33(8):745-753
X-box binding protein1 (XBP1) is an important transcription factor, which participates in many signal transduction procession.To investigate the biological function of XBP1, yeast two-hybrid system to screen proteins interacting with XBP1 in hepatocytes was performed. The XBP1 coding sequence was amplified by polymerase chain reaction (PCR) method, and was cloned in pGEM-T vector. After the target region was sequenced, it was subcloned into the bait plasmid pGBKT7, then was transformed into yeast AH109(a type). After the expression of bait plasmid pGBKT7-XBP1 in AH109 yeast strains were proved by Western blot. The transformed yeast AH109 was mated with yeast Y187(α type) containing hepatocyte cDNA library plasmids pACT2 in 2 ×YPDA medium. Diploid yeast was plated on synthetic dropout nutrient medilium (SD/-Trp-Leu-His-Ade) containing X-α-gal for selection and screening. After sequencing and verifying ORF of positive colonies,7 different kinds of proteins were obtained. In order to further testify the interaction between the screened proteins and XBP1, one of positive colonies MT1E was cloned. The interaction between MT1E and XBP1 in vitro/in vivo were examined successfully with GST pulldown and coimmunoprecipitations. It was shown that MT1E would be a new regulatory protein of XBP1. These screened proteins by yeast two-hybird system were closely correlated with liver fundamental metabolism,protein synthesis and transport,cell proliferation and apoptosis. The results mentioned above contributed to reveal the XBP 1 biological function, and brought some new clues for further exploration of the expressing and regulating mechanism of XBP1.
5.Effects of TGF-?_1 and signal protein Smad3 on rat cardiac myocyte hypertrophy
Jun HUANG ; Fangzhou CHENG ; Junming LI ; Yexin MA
Chinese Journal of Pathophysiology 1989;0(05):-
AIM: To investigate the effects of TGF-?_1 and signal protein Smad3 on rat cardiac myocyte hypertrophy.METHODS: The total protein was analyzed by flow cytometry and the ANF mRNA expression was measured by RT-PCR to judge the hypertrophy of cultured neonatal cardiac myocytes.Smad3 mRNA expression in cardiac myocytes was measured by RT-PCR,and the protein expression of Smad3 was analyzed by Western blotting.RESULTS: TGF-?_1 significantly increased the total protein in cardiac myocytes and promoted ANF mRNA expression,compared with control group.In cultured neonatal myocytes,AS-ODN of Smad3 inhibited myocyte hypertrophy induced by TGF-?_1.Smad3 mRNA and protein expression increased at 15 min after incubated with TGF-?_1,reached the peak at 1 h,and declined at 4 h.CONCLUSION: TGF-?_1 and signal protein Smad3 may participate in the progress of rat cardiac myocyte hypertrophy.
6.Inhibition effect of silencing HPV16 E6 gene on human cervical cancer CaSki cell carcinoma xenograft in nude mice
Haien CHENG ; Yi ZHANG ; Cuili ZHANG ; Weiwei PAN ; Hua SHI ; Faping YI ; Fengjin GUO ; Fangzhou SONG
Journal of Third Military Medical University 1988;0(06):-
Objective To learn the effect on carcinoma xenograft in nude mice by inhibiting human papillomavirus 16(HPV16) E6 gene expression in CaSki cell. Methods The recombinant plasmids expressing HPV16 E6 small interference RNA (siRNA) were transfected into CaSki cell. The cells expressing recombinant plasmid was screened out with G418. The expression of E6 mRNA was determined by RT-PCR. The cells were inoculated into BALB/c nude mice subcutaneouly and the growth of the xenograft carcinoma was observed. After the pGensil-CH2 recombinant was injected into the carcinoma, the growth of carcinoma and pathological changes of carcinoma were observed. Results The CaSki cell expressing E6 siRNA was obtained, and HPV16 E6 mRNA expression in CaSki cell was down-regulated. The oncogenicity of the CaSki cell expressing E6 siRNA was degraded, the inhibition rate was up to 71.4% as compared with that of control group. The growth of tumor in nude mice was inhibited after the E6 siRNA plasmids were injected into the nude mice. The volume and weight of the tumor treated by siRNA were smaller than that of control group significantly. More necrotic area and less cell division phase were observed under light microscope in the E6 siRNA treated tumor. Conclusion The oncogenicity of the CaSki cell was degraded after silencing HPV16 E6 gene in CaSki cell by E6 siRNA.
7.Effects of norepinephrine preconditioning and ischemic preconditioning on apoptosis and Bcl-2, Bax expression in rat myocardial cells during myocardial ischemic reperfusion
Fangzhou CHENG ; Guohua TANG ; Gengshan LI ; Xiqiu YU ; Yuanzhong LI ; Xinhua HE ; Cuiyu BAO ; Yu ZHOU
Chinese Journal of Pathophysiology 2000;0(08):-
AIM: To study the effects of norepinephrine preconditioning(NE-P) and ischemic preconditioning (IP)on apoptosis and Bcl-2, Bax expression in rat myocardial cells in myocardial ischemic reperfusion (I/R). METHODS: The model of rat ischemic-reperfusion was used to conduct NE-preconditioning. Apoptotic myocytes were detected with TUNEL. Bcl-2, Bax expression were detected with immunohistochemistry. RESULTS: The rate of apoptosis cells in I/R group was higher, the rate of apoptosis cells in NE-P group and IP was lower significantly than that in I/R group( P0.05). CONCLUSION: NE-P reduced myocyte apoptosis by I/R in rats; The expression of Bcl-2 ,Bax genes played an important role in myocardial apoptosis.
8.Contamination of Staphylococcus aureus in food sold in Jiading District, Shanghai from 2021 to 2023
Peichao CHEN ; Fangzhou CHENG ; Qiang HUANG ; Huijuan CHEN ; Pan SUN ; Yuting DONG ; Qian PENG
Shanghai Journal of Preventive Medicine 2024;36(7):644-649
ObjectiveTo investigate the contamination status of Staphylococcus aureus in food and the presence of enterotoxin genes in Jiading District, Shanghai, and to provide a basis for the prevention and treatment of foodborne Staphylococcus aureus disease. MethodsFrom 2021 to 2023, 15 types of food were sampled for S. aureus testing, and the presence of five enterotoxin genes, including sea⁃see, was tested in the strains. ResultsOut of 705 food samples, 88 (12.48%) were positive for S. aureus. S. aureus was detected in 12 of the 15 food types, with the three food types with the highest positive rates being cold noodles (45.00%), raw poultry (26.25%), and vegetable salads (20.00%). The enterotoxin gene carriage rate was 32.95% in food strains. The carriage rates for sea, seb, and sec were 7.95%, 12.50%, and 14.77%, respectively. Neither sed nor see was detected. The detection rate of strains carrying two types of enterotoxin genes was 2.27%. The enterotoxin carriage rates in strains from vegetables, beverages, and raw meat were 57.14%, 40.00%, and 30.00%, respectively. ConclusionThe S. aureus detection rate in food in Jiading District is much higher than the national average. The enterotoxin gene carriage rates are high, with food strains carrying sea, seb, and sec, with sec being the most prevalent. There is a need to enhance monitoring of S. aureus and enterotoxins, especially in high-risk foods such as noodles, vegetables, and non-packaged beverages.