1.Effects of MiR-21 on Exercises-induced Right Ventricularfibrosis
Zhijian RAO ; Yun CHANG ; Shiqiang WANG
Chinese Journal of Sports Medicine 2017;36(7):571-577
Objective To explore the expression level of miR-21 in exercise-induced right ventricular (RV) fibrosis,and to analyze the role of miR-21 in exercise-induced right ventricular fibrosis.Methods Seventy-two male Sprague-Dawley rats were randomly divided into a sedentary (Sed) group,a moderate exercise (ME) group and an intensive exercise (IE) group,each of 24.Rats in the Sed group were free of exercises,while those in ME and IE groups ran an hour on treadmill at 5°and 10° slopes at the speed of 15.2 m/min and 28 m/min respectively for 8 weeks,12 weeks or 16 weeks every day,5days per week.Twenty-four hours after the last training,all rats were sacrificed after blood sampling.The right ventricles were removed,and the collagen volume fraction (CVF) was tested using Sirius red staining,Collagen Ⅰ (Col Ⅰ) content was quantified using Immunofluorescence.The expression level of miR-21 was measured using the reverse transcription-polymerase chain reaction (RT-PCR).Results Af ter 12 weeks (P<0.05,P<0.05) and 16 weeks (P<0.01,P<0.01) of intensive exercises,the average CVF in the right ventricular was significantly higher than that of Sed and ME rats.Compared to Sed and ME groups,12 weeks (P<0.01,P<0.01) and 16 weeks (P<0.01,P<0.01) of intensive exercises significantly increased RV collagen Ⅰ content.Compared to the Sed group,the expression of miR-21 in RV increased significantly in the IE group (P<0.01,P<0.05 and P<0.05).After 16-week intensive exercises,the miR 21 expression was positively correlated with the RV Col Ⅰ content.Conclusion The right ventricular fibrosis induced by long-term intensive exercises is associated with increased miR-21 expression level.Therefore,miR-21 is a potent therapeutic target and novel biomarker of the exercise-induced right ventricular fibrosis.
2.Research progress in exosomes in tissue fibrosis
Zhijian RAO ; Yun CHANG ; Shiqiang WANG
Chinese Journal of Pharmacology and Toxicology 2017;31(3):262-268
Exosomes are extracellular nanovesicles secreted by a variety of cell types such as cardiomyocyte, hepatocytes, and stem cells. They carry specific sets of mRNA, microRNA, and proteins, which play a role in intercellular communication in almost each physiological and pathological process. Exosomes, which are released after tissue cell injury, can initiate repair/regeneration responses by triggering inflammation and active fibroblast, and finally lead to tissue fibrosis. However, exosomes released by stem cells can retard tissue fibrosis by enhancing cell survival and reducing apoptosis. In this paper, we reviewed the research progress in the relationship between exosomes and tissue fibrosis.
3.A Comparative Study on Effects of Exercise Intensity and Duration on Left and Right Ventricular
Zhijian RAO ; Yun CHANG ; Shiqiang WANG
Chinese Journal of Sports Medicine 2017;36(2):111-121,105
Objective To examine the effects of exercise intensity and duration on the structure,function and fibrosis of the left and right ventricular,and to discuss the potential mechanism in these processes.Methods Forty-eight male Sprague-Dawley rats were randomly divided into a sedentary(Sed)group,a moderate exercise(ME)group and an intensive exercise(IE) group,each of 16.Rats in Sed group were not given any training,while those in ME group and IE group run on treadmill at the speed of 15.2 m/min with the slope gradient of 5° and 28 m/min with the slope gradient of 10 degree 1 hour per day,5 days per week.Eight and 16 weeks after the training,we recorded the body weight and measure end-diastolic diameter,end-diastolic wall thickness,and ejection fraction of both ventriculars using the ultrasonic testing.All rats were then sacrificed after blood sampling.Elisa was used to measure serum cTnI concentration,and sirius red staining was applied to evaluate collagen volume fraction of both ventriculars.Results Eight or sixteen weeks after the training,the average bi-ventricular end-diastolic diameter of ME and IE rats was bigger than Sed group.There were no differences in end-diastolic diameter of both ventricular between ME group and IE group after sixteen-week training,but the left ventricular end-diastolic diameter of IE group was greater than ME group.As exercise intensive increased and time accumulated,the end-diastolic wall thickness of both ventriculars increased but without statistical significance.At sixteen-week intervention,the bi-ventricular ejection fraction of IE rats was significantly lower than Sed and ME groups,while there was a decreasing trend eight weeks earlier without significant differences.After 8 or 16 weeks of training,the serum cTnI was significantly higher in IE rats than Sed group or ME group,but there was no significant differences between ME group and Sed group.After 16 weeks' exercises,the average bi-ventricular collagen volume fraction of ME or IE group was greater than that after 8 weeks' exercises.The average collagen volume fraction of the right ventricular was greater than Sed group at the same time points,and after sixteenweek training the right ventricular collagen volume fraction in IE group was significantly greater than ME group.However,there were no significant differences in the measurement of the left side among different groups.The serum cTnI was negatively correlated with the left and right ventricular systolic function(r=-0.327,P=0.029 and r=-0.582,P=0.000).Moreover,it was positively correlated with the right ventricular collagen volume fraction moderately,but had no correlation with the left ventricular collagen volume fraction.Conclusion(1)Sixteen-week moderate and intensive exercise result in left ventricular dilation,and the dilation increases with the increase of the exercise intensity.Only 8 weeks' exercise at the same intensity can lead to right ventricular dilation,but exercise intensity has little influence on the right ventricular dilation.(2)Long-term moderate or intensive endurance exercises may cause bi-ventricular hypertrophy potentially.The left ventricular hypertrophy and dilation may not be synchronous with hypertrophy followed by dilation,while the right ventricular hypertrophy and dilation is synchronous.(3)The temporary decrease in bi-ventricular systolic function after intensive endurance exercise may be caused by ventricular injury,with more serious injury in the right ventricular than in the left.Moderate exercises don't cause ventricular injury,thus there is little or no influence on ejection fraction.(4) Long-term (8 or 16 weeks)moderate or intensive endurance exercises can increase the right ventricular collagen volume fraction,which may indicate cardiac fibrosis following right ventricular injury but not in the left ventricular.The bi-ventricular collagen volume fraction at sixteenth week in ME and IE rats are greater than corresponding rats at eighth week.It may result from the hypertrophy of bi-ventricular cardiomyocyte after 8-week training,followed by increase in the extracellular matrix but not cardiac fibrosis.
4.Study on the activation effects to T lymphocytes by gastric cancer cell-dendritic cell fusion vaccines
Kun ZHANG ; Peiwu YU ; Pengfen GAO ; Yun RAO
Chinese Journal of General Surgery 1993;0(02):-
Objective To study the activation effects to cytotoxic T lymphocytes by gastric cancer cell-dendritic cell(DC) fusion vaccines and to provide theoretical data for biological therapy of gastric cancer based on(anti-tumor) fusion vaccines.Methods(1)Peripheral blood mononuclear cells from gastric cancer patients were separated and co-cultured with granulocyte macrophage colony stimulating factors,interleukin-4 and tumor necrosis factor-? to generate mature dendritic cells.(2)The dendritic cells and SGC7901 cells were fusioned by use of polyethylene glycol,and the pure fusion cells were screened out and cultured with HAT and HT(selective) culture systems.(3)The ability of fusion cells to activate the cytotoxic T lymphocytes was(investigated) by using MLA method.Results(1)Mature dendritic cells were gained from gastric cancer(patients)′ peripheral blood mononuclear cells by co-culturing with granulocyte-macrophage colony stimulating factors,interleukin-4 and tumor necrosis factor-?.(2)Dendritic cells and SGC7901 cells could be fusioned by PEG and could get pure fusion cells by culture with HAT/HT selective culture systems.(3)The fusion vaccines could strongly activate cytotoxic T lymphocytes,and there were significent differences between the(fusion) vaccine group and the control group.(P
5.Study on the induction culture and phenotypes of dendritic cells originating from peripheral blood of gastric cancer patients
Kun ZHANG ; Peiwu YU ; Pengfen GAO ; Yun RAO
Chinese Journal of General Surgery 1993;0(03):-
Objective To analyse the morphological and phenotypic characteristics of dendritic cells obtained by induction of peripheral blood monocytes of gastric cancer patients and provide an experimental basis of (dendritic) cells for the biological treatment of gastric cancer. Methods Peripheral blood monocytes of gastric cancer patients were co-cultured with GM-CSF, IL-4 and TNF-? to obtain the dendritic cells. The (morphological) characteristics of the dendritic cells were observed by light and electron microscopes and the (phenotypes) were studied by flow cytometry. Results Mature dendritic cells were oblained seven days after (induction) culture. These cells showed numerous typical dendritic protuberances on their surface.The (phenotypes) of the cells increased gradually with their maturation and reached stable levels seven days later. Conclusions Mature dendritic cells could be obtained from peripheral blood monocytes of gastric cancer (patients) after induction culture with GM-CSF,IL-4 and TNF-?. The obtained cells had typical morphological and phenotypic characteristics. These cells could be used for further research in the biological treatment of (gastric) cancer.
6.A study on the biological characters of SGC7901 gastric cancer cell-dendritic cell fusion vaccine
Kun ZHANG ; Pengfen GAO ; Peiwu YU ; Yun RAO ; Lixin ZHAO
Chinese Journal of General Surgery 1997;0(06):-
Objective To provide experimental basis for the study of SGC7901 gastric cancer cell-dendritic cell fusion vaccine. Methods The fusion of SGC7901 gastric cancer cells and dendritic cells was induced by PEG. We obtained pure fusion cells by selective culture with HAT/HT culture systems. The growth curve characters, clone abilities, abilities to irritate T lymphocytes and the ability to grow were detected. Results The fusion cells had weaker proliferative abilities and clone abilities with increased abilities to irritate T lymphocytes. The CPM value of T lymphocytes were 15083?231、 9608?83、 4214?135、 3020?28 respectively when they were co-cultured with the fusion cells on different proportions. Conclusion SGC7901 gastric cancer cell-dendritic cell fusion cells keep strong abilities to irritate T lymphocytes while inable to grow into new planted tumors in immunodeficiency animals.
7.Experimental study of dendritic cells inducing immunoeffects on anti-gastric cancer
Xiao LEI ; Yan SHI ; Peiwu YU ; Yun RAO
Chinese Journal of General Surgery 1997;0(04):-
Objective To investigate the anti tumor effects of cytotoxic T lymphocytes induced by tumor antigen specific dendritic cells. Methods Soluble tumor antigen was prepared by four freeze thaw cycles of gastric tumor cell line SCG7901. Anti gastric tumor vaccine was acquired by co incubation of tumor antigen and mouse bone marrow derived dendritic cells and cultured with granulocyte/macrophage colony stimulating factor and interleukin 4. Antigen specific cytotoxic T lymphocytes were induced by this tumor vaccine from the spleen and were applied to tumor bearing nude mice. Results Tumor growth was significantly inhibited and the apoptosis of tumor cells was promoted extensively. Conclusions Antigen specific dendritic cell tumor vaccine may play an important role in future immunotherapy of gastric cancer.
8.Chemical constituents of Aconitum brachypodum from Dong-Chuan area.
Hong-Yun WANG ; Ai-Xue ZUO ; Yun SUN ; Gao-Xiong RAO
China Journal of Chinese Materia Medica 2013;38(24):4324-4328
Aconitum brachypodum is traditionally known to be toxic chinese medicie, but its chemical constituents is not enough studied to date. To further elucidate the chemical constituents of A. brachypodum, 80% ethanol extract of A. brachypodum collected from Dong-Chuan area was investigated, which led to isolation of seventeen compounds. By spectroscopic methods, their structures were determined as hypaconitine (1), mesaconitine (2), talatisamine (3), neoline (4), fuziline (5), aconine (6), bullatine A (7), lepeine (8), songrine (9), isocorydine (10), beta-sitosterol (11), daucosterol (12), stearic acid (13), triacontanol (14), palmitic acid (15), benzoic acid (16), and inosine (17), respectively. All compounds except for compounds 1 and 7 were isolated from A. brachypodum for the first time.
Aconitum
;
chemistry
;
China
;
Drugs, Chinese Herbal
;
chemistry
;
Magnetic Resonance Spectroscopy
9.Expressions and clinical significance of CD83 and S100 on gastric cancer infiltrating dendritic cells
Xiao LEI ; Peiwu YU ; Yan SHI ; Bo TANG ; Yun RAO ; Miao WU ; Lixin ZHOU
Chinese Journal of General Surgery 2001;0(10):-
Objective To investigate the expression and clinical significance of tumor infiltrating dendritic cells(TIDCs) within gastric tumor tissues.Methods Immunohistochemistry(IHC),in-situ hybridization(ISH) and flow cytometry were applied to detect the expression of S100,CD83 mRNA and CD83 on DCs in 45 gastric adenocarcinoma tissues.The co-relationship of the S100 and CD83 expression with clinical(pathological) features was analyzed.Results IHC showed that S100 expression was unevenly distributed(within) 42 cancer tissue and CD83 was only expressed in tumor-adjacent tissue and normal tissue.ISH showed that CD83 mRNA was limitedly expressed within 7 samples.S100 expression had no significant(correlation) with clinical pathological features,while CD83 reversely correlated with TNM stages.Detected by flow cytometry,CD83 was expressed in low level in all 45 gastric cancer tissues and negative correlations were found with lymph node metastasis and TNM stages of gastric cancer(r=-0.879,P
10.Effects of CO2 pneumoperitoneum on cell cycle and cell cycle protein of a gastric cancer cell line
Yingxue HAO ; Hua ZHONG ; Peiwu YU ; Chao ZHANG ; Dongzhu ZENG ; Yan SHI ; Yun RAO
Chinese Journal of General Surgery 2010;25(5):389-392
Objective To investigate the effects of different CO2 pneumoperitoneum on cell cycle and cell cycle protein of a gastric cancer cell line. Methods Human gastric cancer cell line MNK-45 were exposed to 0,10,12 and 15 mm Hg CO2 pneumoperitoneum in vitro for 4 hrs. Cell cycle was measured by flowcytometry, the expression of CDK4 ,Cyclin D1、Rb and pRb was studied by Western-blot, and the binding ability of CDK4 and Cyclin D1 was evaluated by immunoprecipitation. Results The cell proliferation index in 15 mm Hg CO2 pneumoperitoneum group dropped significantly (27.4% ± 3. 7%) vs. (36. 4% ±3. 3%) ,P <0. 05, while that in other groups did not change significantly. The protein of CDK4、Cyclin D1and binding ability of Cyclin D1 and CDK4 dropped dramatically in the 15 mm Hg CO2 pneumoperitoneum group (0.71%±0.12%),(0.93% ±0.21%),(0.54%±0.11%),(0.18% ±0.02%) vs. (1.05% ±0.16%),(1.40% ±0.24%),(0.75% ±0.14%),(0.31% ±0.02%), all P<0.05. There were no changes of Rb in protein levels, while the phosphorylated Rb dropped obviously. Conclusion There was no obvious effects of clinical CO2 pneumoperitoneum on gastric cancer cells growth and proliferation.