1.Establishment of a WHBE rabbit knee osteoarthritis model and therapeutic effect of platelet-rich fibrin releasates
Junjie HUANG ; Minli CHEN ; Keyan ZHU ; Jingao JIANG ; Jinghui ZHANG ; Chao JIANG ; Yongming PAN
Chinese Journal of Comparative Medicine 2024;34(5):65-73
		                        		
		                        			
		                        			Objective To investigate the mechanism of cartilage injury and inflammation in the WHBE rabbit KOA model and the effect of platelet-rich fibrin releasates(PRFr)treatment on the KOA process,we established a WHBE rabbit KOA model by excision of medial collateral and partial patellar ligaments and administered a PRFr solution.Methods Twenty-four WHBE rabbits were randomly divided into three groups:normal control(NC)group(n=6),model(KOA)group(n=12),and cure(PRFr)group(n=6).KOA and PRFr groups were injected with 0.5 mL saline and PRFr into both joint cavities on 7 and 14 postoperative days,respectively.At 4 and 8 weeks of modeling,the knee joint grade scoring,X-ray imaging,and gross scoring were performed.Serum levels of IL-1β,TNF-α,and MMP-13 were measured by ELISA.At 4 weeks,6 animals in the KOA group were euthanized,and at 8 weeks,the remaining animals in each group were euthanized.Pathological sections were prepared after decalcification,and then HE,toluidine blue,and safranin O-fast green staining and immunohistochemical analysis of TGF-β,BMP3,and NF-κB were conducted.Results The Lequesne MG behavioral score,Mankin's score,and Pelletier score of WHBE rabbits after the operation were significantly increased compared with the NC group(P<0.01).Pathological observations revealed surface defects of the cartilage and partial loss of chondrocytes.These result indicated that the KOA model was established successfully.In KOA rabbits,knee joint swelling,joint pain stimulation,and movement limitation were obvious.X-rays showed a high-density soft tissue shadow,indicating more joint effusion and a rough articular surface in general.After PRFr treatment,the serum levels of proinflammatory factors IL-1β,TNF-α,and MMP-13 in KOA model rabbits were significantly reversed(P<0.05,P<0.01).Additionally,the cartilage surface became smooth,and most chondrocytes were neatly distributed.Expression levels of TGF-β,BMP3,and NF-κB induced by KOA were also significantly decreased(P<0.01).Conclusions We successfully established a KOA model in WHBE rabbits,and PRFr improved the cartilage injury and inflammation of the WHBE rabbit KOA model through TGF-β/BMP and NF-κB pathways.
		                        		
		                        		
		                        		
		                        	
2.Study on the protective effect of Guanxinning Tablet on myocardial ischemia reperfusion injury in rats
Fei HU ; Yili RONG ; Keyan ZHU ; Hong LU ; Cheng CHEN ; Minli CHEN ; Yongming PAN
Chinese Journal of Comparative Medicine 2017;27(5):76-82
		                        		
		                        			
		                        			Objective To observe the effect of Guanxinning Tablet (GXNT) on myocardial infarction and cardiac autonomic nervous function in rats with myocardial ischemia reperfusion injury (MI/RI).Methods Seventy SD rats were divided into 7 groups randomly (n=10);the sham group, the MI/RI group, 75 mg/kg, 150 mg/kg, 300 mg/kg and 600 mg/kg GXNT groups and 300 mg/kg Compound Danshen Tablets (DST) group.All rats were administered orally for 7 days, and then the MI/RI model was made by ligating the left anterior descending branch of coronary artery in rats.The changes of electrocardiogram were recorded and the electrocardiogram of J points and heart rate variability (HRV) parameters were analyzed.At the end of reperfusion, the myocardial infarct size was measured by using Evans blue and tetrazolium chloride (TTC) double staining, and pathological changes of myocardium were observed by HE staining.The changes of serum lactate dehydrogenase (LDH), creatine kinase (CK), malondialdehyde (MDA) and nitric oxide (NO) levels were also detected.Results Compared with MI/RI group, GXNT and DST groups were significantly reduced myocardial infarct size and inhibited the rising of serum LDH and CK activities (P < 0.05, P < 0.01), and also reduced the total or average value of J point during reperfusion (P < 0.05, P < 0.01).Meanwhile, GXNT and DST groups were markedly increased HRV and serum NO level as well as decreased serum MDA content (P < 0.05, P < 0.01), and improved myocardial tissue pathology.Conclusions GXNT can reduce the myocardial infarction in rats with MI/RI, and also improve the cardiac autonomic nervous function.
		                        		
		                        		
		                        		
		                        	
3.Establishment of chronic myocardial ischemia model in minipigs and application of noninvasive telemetry technique
Minli CHEN ; Yongming PAN ; Liang CHEN ; Xiaoping XU ; Dejun WANG ; Yuwei YANG ; Fangming CHEN ; Yueqin CAI ; Jianqin XU ; Keyan ZHU
Chinese Journal of Comparative Medicine 2017;27(5):16-18
		                        		
		                        			
		                        			In order to establish the minipig model of chronic myocardial ischemia and apply non-invasive telemetry technique, the minipig model of chronic myocardial ischemia was made induced by Vitamin D3, isoproterenol and combined with high fat diet, and the non-invasive telemetry technique was used to detect and evaluate the symptoms of minipig model of chronic myocardial ischemia.Moreover, the effects of transport stress and the risk factors of atherosclerosis (AS) induced by high fat diet among Wuzhishan minipigs, Bama minipigs and Tibetan minipigs were also evaluated.Our study has successful established the Bama minipig model of chronic myocardial ischemia and the technical specification for evaluation,.The non-invasive telemetry technique can be used to detect and evaluate the symptoms of chronic myocardial ischemia model, and defines minipigs at least need to keep for more than 4 weeks after transport stress to adaptive recovery period.In addition, the different characteristics of AS risk factors such as hyperlipidemia, hypertension and hyperinsulinemia were observed in Wuzhishan minipigs, Bama minipigs and Tibetan minipigs in high fat environment, and this provides a reference for the selection and application of minipigs in the research of cardiovascular diseases.
		                        		
		                        		
		                        		
		                        	
4.Application of enhanced recovery after surgery in the perioperative period of patients undergoing hepatectomy
Yuanpeng WANG ; Huichun LIU ; Wei HUANG ; Keyan PAN ; Hao JIN ; Lei ZHOU
Chinese Journal of General Surgery 2017;32(3):243-246
		                        		
		                        			
		                        			Objective To observe the safety and clinical efficacy of enhanced recovery after surgery in the perioperation period of hepatectomy.Methods 92 patients with primary hepatic cancer who underwent surgical resection in our hospital from December 2014 to May 2016 were divided into two groups,50 patients received well-organized and consecutive clinical interventions guided by ERAS.42 patients underwent traditional perioperative management.Results Compared with traditional group,ERAS group had reduced hospital cost [(40 633.12 ± 6 336.46) RMB vs.(46 139.23 ± 9 605.88) RMB,P < 0.05],shorter postoperative hospital stay[(10.24 ± 1.6) d vs.(13.35 ± 4.86) d,P < 0.05],earlier flatus and defecation [(33.34 ±6.01) h vs.(50.31 ± 3.53) h,P < 0.05],and improved satisfaction rate for pain management[45/50(90%) vs.22/42(52.4%),P < 0.05];while the postoperative adverse events and complications of the two groups showed no difference (P > 0.05).Conclusion The application of ERAS in the perioperation period of hepatectomy is safe and effective,reducing hospital cost,postoperative hospital stay,improving satisfaction rate to pain management,and facilitating recovery in hepatic surgery.
		                        		
		                        		
		                        		
		                        	
5.Observation on the sensitivity of Wuzhishan, Tibetan and Bama minipigs to exogenous fats
Liang CHEN ; Yongming PAN ; Keyan ZHU ; Xiaoping XU ; Jianqin XU ; Minli CHEN
Chinese Journal of Comparative Medicine 2017;27(6):12-16
		                        		
		                        			
		                        			Objective To observe the sensitivity of Wuzhishan, Tibetan and Bama minipigs to exogenous fats.Methods A total of 15 male minipigs including 5 WZS minipigs, 5 Tibetan minipigs and 5 Bama minipigs, were used in this study.The minipigs were intravenously injected with fat emulsion and fed with high-fat diet, and the changes of serum total cholesterol (TC), low density lipoprotein (LDL-C), high density lipoprotein (HDL-C) and triglyceride (TG) levels were detected at 0.5 h, 1 h, 1.5 h, 2 h, 2.5 h and 3 h after intravenous injection of fat emulsion and at 3 h and 12 h, 24 h, 36 h, 48 h, 60 h, 72 h and 84 h after fed with high-fat diet, respectively, and calculated the changes of area under the curve (ΔAUC) values.Results The triglyceride (TG) in the three kinds minipigs were significantly increased (P<0.01), and reached a peak at 0.5 h after injection.The degree of elevation of ΔAUC values were different showing on increasing order: Tibetan minipig > Wuzhishan minipig > Bama minipig, while TC, LDL-C and HDL-C showed no significant changes (P>0.05).Moreover, TC, LDL-C, HDL-C and TG were significantly increased in the three stocks of minipigs induced by feeding with high-fat diet (P<0.05, P<0.01).Among them, the TC and LDL-C of Wuzhishan minipigs peaked at 36 h, HDL-C peaked at 48 h and TG peaked at 24 h after feeding, respectively.TC, LDL-C and HDL-C in the Bama minipigs and Tibetan minipigs peaked at 48-60 h and TG peaked at 36 h after feeding, and the ΔAUC values of TC, LDL-C and HDL-C were in an increasing order of Wuzhishan minipigs > Bama minipigs > Tibetan minipigs.Conclusions The three stocks of minipigs are sensitive to TG after intravenous injection of fat emulsion, and the lipid tolerance values are in an order of Tibetan minipig > Wuzhishan minipigs > Bama minipigs.Meanwhile, the three stocks of minipigs are also sensitive to TC, LDL-C and HDL-C after feeding with high-fat diet, and the lipid tolerance values are in an increasing order of Wuzhishan minipig > Bama minipigs > Tibetan minipigs.
		                        		
		                        		
		                        		
		                        	
6.Effect of sevoflurane preconditioning on brain injury induced by cardiopulmonary bypass in rats
Nan ZHOU ; Yifei PAN ; Keyan CHEN ; Tiezheng ZHANG ; Jin ZHOU
Chinese Journal of Anesthesiology 2016;36(2):165-167
		                        		
		                        			
		                        			Objective To evaluate the effect of sevoflurane preconditioning on brain injury induced by cardiopulmonary bypass (CPB) in rats.Methods Forty adult male Sprague-Dawley rats,aged 6-8 months,weighing 350-450 g,were randomly divided into 5 groups (n=8 each) using a random number table:sham operation group (S group),CPB group,and preconditioning with different concentrations of sevoflurane groups (SP1,SP2 and SP3 groups).In SP1,SP2 and SP3 groups,sevoflurane with the final concentrations of 1.2%,2.4% and 3.6%,respectively,was inhaled for 1 h,and then CPB was started.After sevoflurane preconditioning and before CPB (T0),at 30 min of CPB (T1),at the end of CPB (T2),and at 1,2 and 3 h after termination of CPB (T3-5),venous blood samples were collected from the right internal jugular vein for determination of serum S100-β protein concentrations by enzyme-linked immunosorbent assay.Rats were sacrificcd at T5,and hippocampi were isolated for determination of neuronal apoptosis (by TUNEL) and NF-κB p65 expression (by immunohistochemistry).Results Compared with group S,the concentration of serum S100-β protein was significantly increased at T1-5,the number of apoptotic neurons was significantly increased,and the expression of NF-κB p65 was significantly up-regulated in CPB,SP1,SP2 and SP3 groups (P<0.05).Compared with group CPB,the serum S100-β protein concentration was significantly decreased at T1-5,the number of apoptotic neurons was significantly decreased,and the expression of NF-κB p65 was significantly down-regulated in SP1,SP2 and SP3 groups (P< 0.05).Compared with group SP1,the serum S100-β protein concentration was significantly decreased at T1-5,the number of apoptotic neurons was significantly decreased,and the expression of NF-κB p65 was significantly down-regulated in SP2 and SP3 groups (P<0.05).Compared with group SP2,the serum S100-β protein concentration was significantly decreased at T1-5,the number of apoptotic neurons was significantly decreased,and the expression of NF-κB p65 was significantly downregulated in group SP3 (P<0.05).Conclusion Sevoflurane preconditioning can attenuate CPB-induced brain injury probably by inhibiting activation of NF-κB in hippocampal neurons of rats.
		                        		
		                        		
		                        		
		                        	
7.Experimental research on the prevention of rabbit postoperative abdominal cavity adhesion with PLGA membrane.
Xiubing PANG ; Yongming PAN ; Fei HUA ; Chaoying SUN ; Liang CHEN ; Fangming CHEN ; Keyan ZHU ; Jianqin XU ; Minli CHEN
Journal of Biomedical Engineering 2015;32(1):146-162
		                        		
		                        			
		                        			The aim of this paper is to explore the prevention of rabbit postoperative abdominal cavity adhesion with poly (lactic-co-glycotic acid) (PLGA) membrane and the mechanism of this prevention function. Sixty-six Japanese white rabbits were randomly divided into normal control group, model control group and PLGA membrane group. The rabbits were treated with multifactor methods to establish the postoperative abdominal cavity adhesion models except for those in the normal control group. PLGA membrane was used to cover the wounds of rabbits in the PLGA membrane group and nothing covered the wounds of rabbits in the model control group. The hematologic parameters, liver and kidney functions and fibrinogen contents were detected at different time. The rabbit were sacrificed 1, 2, 4, 6, 12 weeks after the operations, respectively. The adhesions were graded blindly, and Masson staining and immunohistochemistry methods were used to observe the proliferation of collagen fiber and the expression of transforming growth factor β1 (TGF-β1) on the cecal tissues, respectively. The grade of abdominal cavity adhesion showed that the PLGA membrane-treated group was significant lower than that in the model control group, and it has no influence on liver and kidney function and hematologic parameters. But the fibrinogen content and the number of white blood cell in the PLGA membrane group were significant lower than those of model control group 1 week and 2 weeks after operation, respectively. The density of collagen fiber and optical density of TGF-β1 in the PLGA membrane group were significant lower than those of model control group. The results demonstrated that PLGA membrane could be effective in preventing the abdominal adhesions in rabbits, and it was mostly involved in the reducing of fibrinogen exudation, and inhibited the proliferation of collagen fiber and over-expression of TGF-β1.
		                        		
		                        		
		                        		
		                        			Abdominal Cavity
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		                        			surgery
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		                        			Animals
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		                        			Collagen
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		                        			metabolism
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		                        			Lactic Acid
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		                        			Polyglycolic Acid
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		                        			Rabbits
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		                        			Tissue Adhesions
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		                        			prevention & control
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		                        			Transforming Growth Factor beta1
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		                        			metabolism
		                        			
		                        		
		                        	
8.Effects of testosterone deficiency on serum lipid levels and hepatic lipid accumulation in miniature pigs fed a high-fat diet
Zhaowei CAI ; Yongming PAN ; Liang CHEN ; Keyan ZHU ; Fangming CHEN ; Yueqin CAI ; Xiaoping XU ; Minli CHEN
Chinese Journal of Comparative Medicine 2015;(1):40-44
		                        		
		                        			
		                        			Objective The aim of this study was to explore the effect of testosterone deficiency on serum lipid levels and hepatic lipid accumulation in miniature pigs fed a high-fat diet (HFD).Methods Eighteen sexually mature male Chinese Wuzhishan miniature pigs (6~7 months old) were used in this study.The pigs were divided in three groups ( n =6 animals/group ) as follows: intact male pigs , castrated male pigs and castrated male pigs with testosterone replacement .They were fed a HFD diet for 12 weeks and body weights were recorded weekly .Serum levels of testosterone , total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C) and triglycerides (TG) were measured.Hepatic TG and TC levels were also determined , and liver tissues were embedded in paraffin and stained with hematoxylin and eosin (H&E).Results (1) The body weights of pigs in each group were found to be linearly elevated over time .Though castrated pigs gained less weight than did pigs in the other groups , no significant differences were found between them .( 2 ) Castration caused a significant decrease in serum testosterone levels in pigs . This effect was recovered by testosterone treatment .(3) Serum levels of TC, LDL-C and TG were significantly increased in castrated pigs.However, castration had no significant effect on serum HDL-C levels.Testosterone treatment reduced the increased serum lipids in castrated pigs .(4) Hepatic TG and TC contents in castrated pigs were also significantly higher than those in other groups of pigs .Testosterone treatment reduced the increased hepatic lipids in castrated pigs .( 5 ) Compared with other groups of pigs , castrated pigs showed increased steatosis .However , testosterone treatment attenuated hepatic steatosis in castrated pigs .Conclusion Testosterone deficiency caused severe dyslipidemia , and increased hepatic lipid accumulation in miniature pigs fed a high-fat diet.
		                        		
		                        		
		                        		
		                        	
9.Establishment of a minipig model of chronic myocardial ischemia of phlegm-blood stasis syndrome type
Yongming PAN ; Liang CHEN ; Xiaoping XU ; Keyan ZHU ; Jianqin XU ; Minli CHEN
Chinese Journal of Comparative Medicine 2015;(9):1-7
		                        		
		                        			
		                        			ObjectiveToestablishadiseasesyndromecombinedanimalmodel,theminiaturepigmodelof chronic myocardial ischemia of phlegm-blood stasis syndrome type , by high fat/cholesterol diet feeding and intravenous injection with VD3 and isoproterenol.Methods Miniature pigs were randomly divided into the control (Ctr) group, high fat/cholesterol diet ( HFC) group and chronic myocardial ischemia model of phlegm-blood stasis syndrome ( CMI) group, 5 pigs in each group .The Ctr group was fed with normal regular chow diet , HFC group was fed with high fat/cholesterol diet , while the CMI group was fed with high fat/cholesterol diet and intravenous injection with VD 3 and isoproterenol .The experiment lasted for 24 weeks.The model establishment and its pathological process of phlegm-blood stasis syndrome were evaluated by examinations of body weight , electrocardiogram, activity, blood lipid, myocardial enzymes, hemorheology, inflammation, cardiac index(CI) and myocardial ischemia size (MIS).Results Compared with the Ctr group, the body weight, heart rate(HR), Total cholesterol(TC), low density lipoprotein-cholesterol (LDL-C), high density lipoprotein-cholesterol ( HDL-C ) , atherosclerosis index ( AI ) , low/middle/high shear rate of whole blood viscosity and reduced viscosity, erythrocyte electrophoresis time(EPT), high-sensitivity C-reactive protein (hs-CRP) and IL-6 levels in the HFC group were significantly increased (P <0.05, P <0.01), while the body weight, heart rate, total ST, ST_average, activity, TC, TG, LDL-C, HDL-C, AI, CK, LDH, cTn-1, low/middle/high shear rate of whole blood viscosity and reduced viscosity, EPT, Casson viscosity(CV), hs-CRP, IL-6, CI and MIS in the CMI group were significantly increased (P<0.05, P<0.01), and APN level in the CMI group was significantly decreased (P<0.05).Moreover, AI, CK, LDH, cTn-1, low/middle/high shear rate of whole blood viscosity , EPT, CI and MIS in the CMI group were significantly higher than those of HFC group (P<0.05, P<0.01), while APN in the CMI group was significantly lower than that of HFC group (P<0.05).Correlation analysis showed that MIS was closely correlated to TC , LDL-C, AI, CK, LDH, cTn-1, APN, high/middle/low shear rate of whole blood viscosity , EPT, CV, hs-CRP and IL-6 (P<0.05, P<0.01).The linear regression analysis also showed that phlegm-blood stasis was closely correlated to TC , LDL-C, AI, CK, LDH, cTn-1, APN, CV, EPT, hs-CRP, and IL-6 ( P <0.01), and further linear stepwise regression analysis showed that the evolution of phlegm-blood stasis was closely related to TC , CK and IL-6.Conclusions Minipig model of chronic myocardial ischemia of phlegm-blood stasis syndrome type can be established by high fat /cholesterol diet feeding and intravenous injection with VD 3 and isoproterenol .Their blood lipid metabolism , hemorheology , myocardial enzymes and inflammatory indexes can be used as external biochemical basis of phlegm-blood stasis syndrome type , which may reflect related biological basis of the traditional Chinese medicine theory of “phlegm and stasis cementation , blood-stasis & toxin causing catastrophe”.
		                        		
		                        		
		                        		
		                        	
10.Effects of PLGA absorbable membrane on preventing postoperative abdominal adhesion in rabbits.
Xiubing PANG ; Yongming PAN ; Fei HUA ; Chaoying SUN ; Liang CHEN ; Fangming CHEN ; Keyan ZHU ; Jianqin XU ; Minli CHEN
Chinese Journal of Medical Instrumentation 2014;38(5):389-392
OBJECTIVETo investigate the effects of PLGA absorbable membrane in prevention of postoperative abdominal adhesion in rabbits.
METHODS66 Japanese white rabbits were randomly divided into three groups: the normal control group n = 6, model control group n = 30 and PLGA group n = 30. Rabbits were received multifactor methods to establish postoperative abdominal adhesion models except for normal control group. The cecum wound was covered PLGA membrane in the PLGA group. At postoperative 1, 2, 4, 6 and 12 weeks, the abdominal cavities were reopened and the adhesive severity was graded blindly, and the hydroxyproline level in cecum tissue was measured and the cecum histopathology was observed.
RESULTS(1) the degree of adhesion and hydroxyproline level in model control group were significantly higher than those of normal control group (P < 0.05, P < 0.01), while the degree of adhesion and hydroxyproline level in PLGA group were significantly lower than those of model control group (P < 0.05). (2) HE staining showed that cecum serosa had obviously inflammatory cell infiltration and fibroblast proliferation, while PLGA could inhibit fibroblast proliferation and reduce the inflammatory cell infiltration and collagen.
CONCLUSIONPLGA absorbable membrane can inhibit fibroblast proliferation and collagen to prevent the experimental postoperative peritoneal adhesions.
Abdominal Cavity ; pathology ; Animals ; Cell Proliferation ; Collagen ; analysis ; Fibroblasts ; cytology ; Lactic Acid ; chemistry ; Membranes, Artificial ; Polyglycolic Acid ; chemistry ; Postoperative Complications ; prevention & control ; Rabbits ; Tissue Adhesions ; prevention & control
            
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