目的:探究4℃冷藏过程中鲈鱼肌肉品质的变化。方法:测定4℃冷藏过程中鲈鱼肌肉pH值、菌落总数、总挥发性盐基氮(total volatile base nitrogen,TVB-N)含量、K值和质构等指标,综合评价鲈鱼生理生化品质的变化,采用Masson染色和免疫组化...目的:探究4℃冷藏过程中鲈鱼肌肉品质的变化。方法:测定4℃冷藏过程中鲈鱼肌肉pH值、菌落总数、总挥发性盐基氮(total volatile base nitrogen,TVB-N)含量、K值和质构等指标,综合评价鲈鱼生理生化品质的变化,采用Masson染色和免疫组化分析评价组织形态学变化,利用酶解荧光肽底物测定特异性酶解胶原蛋白的基质金属蛋白酶(matrix metalloproteinases,MMPs)活力变化。结果:鲈鱼在贮藏期间pH值呈先下降后升高趋势,菌落总数、TVB-N含量和K值随贮藏时间的延长而增加,并在第8天超过临界值。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析结果显示,贮藏至6 d鲈鱼肌肉蛋白组成没有明显变化,而在第8天肌球蛋白重链降解明显。Masson染色和I型胶原免疫组化分析结果显示,随贮藏时间的延长,肌内膜逐渐被降解,至第12天,肌膜I型胶原蛋白和肌纤维之间出现明显间隙。MMPs活力随贮藏时间的延长而明显增加,而鱼肉硬度不断下降。结论:鱼类冷藏期间肌肉的软化与胶原蛋白分解密切相关,而MMPs对I型胶原蛋白的降解可能是鱼死后肌肉软化的主要原因。展开更多
目的:观察扶正利湿解毒法基本方(简称基本方)对肝纤维化的逆转作用.方法:采用DMN诱导大鼠肝纤维化,造模结束后随机分为模型对照组,秋水仙碱组,基本方高剂量组,基本方中剂量组及基本方低剂量组并分组给药,同时设空白对照组.灌胃3wk后检...目的:观察扶正利湿解毒法基本方(简称基本方)对肝纤维化的逆转作用.方法:采用DMN诱导大鼠肝纤维化,造模结束后随机分为模型对照组,秋水仙碱组,基本方高剂量组,基本方中剂量组及基本方低剂量组并分组给药,同时设空白对照组.灌胃3wk后检测肝组织Hyp,ALT水平;HE,Masson染色.采用HSC-T6细胞培养,药物作用24h后光镜下观察拍照,检测药物对细胞的抑制率并在电镜下观察药物浓度为2g/L时细胞的变化.结果:基本方高、中剂量均可使大鼠肝组织Hyp,血清ALT显著下降(Hyp:213.73±58.05μg/g,245.95±67.70μg/g vs 317.37±42.06μg/g,P<0.01,P<0.05;ALT:33.56±8.16 kU/L,42.38±14.99 kU/L vs 58±10.32 kU/L,P<0.01,P<0.05);Masson染色肝纤维化分级评分显示,基本方高,中剂量均可使肝纤维化程度显著降低(2.33±0.87,2.63±0.74 vs 3.70±0.67,P<0.01,P<0.01),同时显著改善肝组织炎症、出血及纤维沉积;光镜观察及抑制率检测显示基本方呈剂量依赖性抑制HSC-T6细胞增值.电镜显示基本方主要导致HSC-T6细胞凋亡.结论:基本方显示了显著的体内外逆转肝纤维化作用.展开更多
Electrical stimulation has been shown to accelerate and enhance nerve regeneration in sensory and motor neurons after injury, but there is little evidence that focuses on the varying degrees of fibrosis in the delayed...Electrical stimulation has been shown to accelerate and enhance nerve regeneration in sensory and motor neurons after injury, but there is little evidence that focuses on the varying degrees of fibrosis in the delayed repair of peripheral nerve tissue. In this study, a rat model of sciatic nerve transec- tion injury was repaired with a biodegradable conduit at 1 day, 1 week, 1 month and 2 months after injury, when the rats were divided into two subgroups. In the experimental group, rats were treated with electrical stimuli of frequency of 20 Hz, pulse width 100 ms and direct current voltage of 3 V; while rats in the control group received no electrical stimulation after the conduit operation. His- tological results showed that stained collagen fibers comprised less than 20% of the total operated area in the two groups after delayed repair at both 1 day and 1 week but after longer delays, the collagen fiber area increased with the time after injury. Immunohistochemical staining revealed that the expression level of transforming growth factor ~ （an indicator of tissue fibrosis） decreased at both 1 day and 1 week after delayed repair but increased at both 1 and 2 months after delayed repair. These findings indicate that if the biodegradable conduit repair combined with electrical stimulation is delayed, it results in a poor outcome following sciatic nerve injury. One month after injury, tissue degeneration and distal fibrosis are apparent and are probably the main reason why electrical stimulation fails to promote nerve regeneration after delayed repair.展开更多
目的:观察白虎加桂枝汤对尿酸性肾病(HN)大鼠的治疗效果,并从肾小管损伤角度探讨其作用机制。方法:将60只无特定病原体(SPF)级SD雄性大鼠随机分为正常对照组、模型组、白虎加桂枝汤高剂量组(BHJGZ高组)、白虎加桂枝汤中剂量组(BHJGZ中组...目的:观察白虎加桂枝汤对尿酸性肾病(HN)大鼠的治疗效果,并从肾小管损伤角度探讨其作用机制。方法:将60只无特定病原体(SPF)级SD雄性大鼠随机分为正常对照组、模型组、白虎加桂枝汤高剂量组(BHJGZ高组)、白虎加桂枝汤中剂量组(BHJGZ中组)、白虎加桂枝汤低剂量组(BHJGZ低组)、别嘌醇组,每组10只。采用腺嘌呤灌胃联合腹部皮下注射氧嗪酸钾构建大鼠尿酸性肾病模型,连续造模18 d。造模成功后,正常对照组及模型组给予等体积蒸馏水灌胃,其余各组给予对应药物灌胃,连续干预8周。观察大鼠体质量、活动能力、精神状态及毛色,检测大鼠血清尿酸(UA)、血清尿素氮(BUN)、血清肌酐(Scr)、24 h尿蛋白定量(24 h UTP),通过HE染色、过碘酸希夫(PAS)染色和Masson染色,观察高尿酸血症(HUA)大鼠肾脏组织形态学、肾小球系膜增生以及肾间质纤维化情况。结果:与模型组比较,治疗6周后,白虎加桂枝汤各剂量组大鼠体质量增加,活动能力、精神状态及毛色好转(P<0.05)。BHJGZ中组可显著降低大鼠血清Scr水平(P<0.01),BHJGZ中组、BHJGZ高组可明显降低大鼠血清BUN水平(P<0.01);白虎加桂枝汤各剂量组均可明显降低HUA大鼠血清UA、24 h UTP水平并改善肾组织形态学、肾小球系膜增生及肾间质纤维。结论:白虎加桂枝汤可能通过抑制肾小球系膜基质增生、减少肾小管损伤及肾间质纤维化,延缓HN进展。展开更多
Polyethylene glycol can connect the distal and proximal ends of an injured nerve at the cellular level through axonal fusion to avoid Wallerian degeneration of the injured distal nerve and promote peripheral nerve reg...Polyethylene glycol can connect the distal and proximal ends of an injured nerve at the cellular level through axonal fusion to avoid Wallerian degeneration of the injured distal nerve and promote peripheral nerve regeneration.However,this method can only prevent Wallerian degeneration in 10% of axons because the cytoskeleton is not repaired in a timely fashion.Reconstruction of the cytoskeletal trunk and microtubule network has been suggested to be the key for improving the efficiency of axonal fusion.As a microtubule-severing protein,spastin has been used to enhance cytoskeletal reconstruction.Therefore,we hypothesized that spastin combined with polyethylene glycol can more effectively promote peripheral nerve regeneration.A total of 120 male Sprague-Dawley rats were randomly divided into sham,suture,polyethylene glycol,and polyethylene glycol + spastin groups.In suture group rats,only traditional nerve anastomosis of the end-to-end suture was performed after transection of the sciatic nerve.In polyethylene glycol and polyethylene glycol + spastin groups,50 μL of polyethylene glycol or 25 μL of polyethylene glycol + 25 μL of spastin,respectively,were injected immediately under the epineurium of the distal suture.Sensory fiber regeneration distance,which was used to assess early nerve regeneration at 1 week after surgery,was shortest in the suture group,followed by polyethylene glycol group and greatest in the polyethylene glycol + spastin group.Behavioral assessment of motor function recovery in rats showed that limb function was restored in polyethylene glycol and polyethylene glycol + spastin groups at 8 weeks after surgery.At 1,2,4 and 8 weeks after surgery,sciatic functional index values and percentages of gastrocnemius muscle wet weight were highest in the sham group,followed by polyethylene glycol + spastin and polyethylene glycol groups,and lowest in the suture group.Masson staining was utilized to assess the morphology of muscle tissue.Morphological changes in skeletal muscle were detectable in sutur展开更多
以O/W型软膏基质作为载体,制备Rhc-Ⅲ(Recombinant human collagenⅢ,简称Rhc-Ⅲ)乳膏剂并对其质量和促创伤愈合作用进行评价。采用乳化法制备Rhc-Ⅲ乳膏剂,BCA法测定其总胶原蛋白的含量,并对其外观性状、pH、黏度、重金属和微生物等进...以O/W型软膏基质作为载体,制备Rhc-Ⅲ(Recombinant human collagenⅢ,简称Rhc-Ⅲ)乳膏剂并对其质量和促创伤愈合作用进行评价。采用乳化法制备Rhc-Ⅲ乳膏剂,BCA法测定其总胶原蛋白的含量,并对其外观性状、pH、黏度、重金属和微生物等进行质量检查;采用切除全层皮肤法建立大鼠伤口模型,以伤口的愈合率和伤口组织切片的HE染色、Masson染色结果为评价指标,对Rhc-Ⅲ乳膏剂促进伤口愈合作用进行药效学评价。乳化法制备的Rhc-Ⅲ乳膏剂外观为乳白色、黏稠、无异味且易于涂布的半固体,其pH为(6.66±0.03),黏度为(48 136±595)mPa·s,总胶原蛋白的含量为(1.38±0.15) mg/g,重金属和微生物限度符合要求;耐寒耐热试验后稳定性良好;模型大鼠给药后在3,7和14 d, Rhc-Ⅲ乳膏剂组的伤口愈合率均大于模型组和空白乳膏组;HE染色和Masson染色结果显示,Rhc-Ⅲ乳膏剂组肉芽组织出现早,成熟快,胶原纤维沉积量多,伤口愈合效果好。Rhc-Ⅲ乳膏剂制备方法简便、质量可控且稳定,具有促进伤口愈合的作用。展开更多
文摘目的:观察扶正利湿解毒法基本方(简称基本方)对肝纤维化的逆转作用.方法:采用DMN诱导大鼠肝纤维化,造模结束后随机分为模型对照组,秋水仙碱组,基本方高剂量组,基本方中剂量组及基本方低剂量组并分组给药,同时设空白对照组.灌胃3wk后检测肝组织Hyp,ALT水平;HE,Masson染色.采用HSC-T6细胞培养,药物作用24h后光镜下观察拍照,检测药物对细胞的抑制率并在电镜下观察药物浓度为2g/L时细胞的变化.结果:基本方高、中剂量均可使大鼠肝组织Hyp,血清ALT显著下降(Hyp:213.73±58.05μg/g,245.95±67.70μg/g vs 317.37±42.06μg/g,P<0.01,P<0.05;ALT:33.56±8.16 kU/L,42.38±14.99 kU/L vs 58±10.32 kU/L,P<0.01,P<0.05);Masson染色肝纤维化分级评分显示,基本方高,中剂量均可使肝纤维化程度显著降低(2.33±0.87,2.63±0.74 vs 3.70±0.67,P<0.01,P<0.01),同时显著改善肝组织炎症、出血及纤维沉积;光镜观察及抑制率检测显示基本方呈剂量依赖性抑制HSC-T6细胞增值.电镜显示基本方主要导致HSC-T6细胞凋亡.结论:基本方显示了显著的体内外逆转肝纤维化作用.
基金supported by grants from the National Program on Key Basic Research Project of China(973 Program),No.2014CB542206Program for Innovative Research Team in University of Ministry of Education of China,No.IRT1201+2 种基金the National Natural Science Foundation of China,No.31271284,31171150,81171146,30971526,31100860,31040043,31440055the Natural Science Foundation of Beijing of China,No.7142164Program for New Century Excellent Talents in University of Ministry of Education of China,No.BMU20110270
文摘Electrical stimulation has been shown to accelerate and enhance nerve regeneration in sensory and motor neurons after injury, but there is little evidence that focuses on the varying degrees of fibrosis in the delayed repair of peripheral nerve tissue. In this study, a rat model of sciatic nerve transec- tion injury was repaired with a biodegradable conduit at 1 day, 1 week, 1 month and 2 months after injury, when the rats were divided into two subgroups. In the experimental group, rats were treated with electrical stimuli of frequency of 20 Hz, pulse width 100 ms and direct current voltage of 3 V; while rats in the control group received no electrical stimulation after the conduit operation. His- tological results showed that stained collagen fibers comprised less than 20% of the total operated area in the two groups after delayed repair at both 1 day and 1 week but after longer delays, the collagen fiber area increased with the time after injury. Immunohistochemical staining revealed that the expression level of transforming growth factor ~ （an indicator of tissue fibrosis） decreased at both 1 day and 1 week after delayed repair but increased at both 1 and 2 months after delayed repair. These findings indicate that if the biodegradable conduit repair combined with electrical stimulation is delayed, it results in a poor outcome following sciatic nerve injury. One month after injury, tissue degeneration and distal fibrosis are apparent and are probably the main reason why electrical stimulation fails to promote nerve regeneration after delayed repair.
文摘目的:观察白虎加桂枝汤对尿酸性肾病(HN)大鼠的治疗效果,并从肾小管损伤角度探讨其作用机制。方法:将60只无特定病原体(SPF)级SD雄性大鼠随机分为正常对照组、模型组、白虎加桂枝汤高剂量组(BHJGZ高组)、白虎加桂枝汤中剂量组(BHJGZ中组)、白虎加桂枝汤低剂量组(BHJGZ低组)、别嘌醇组,每组10只。采用腺嘌呤灌胃联合腹部皮下注射氧嗪酸钾构建大鼠尿酸性肾病模型,连续造模18 d。造模成功后,正常对照组及模型组给予等体积蒸馏水灌胃,其余各组给予对应药物灌胃,连续干预8周。观察大鼠体质量、活动能力、精神状态及毛色,检测大鼠血清尿酸(UA)、血清尿素氮(BUN)、血清肌酐(Scr)、24 h尿蛋白定量(24 h UTP),通过HE染色、过碘酸希夫(PAS)染色和Masson染色,观察高尿酸血症(HUA)大鼠肾脏组织形态学、肾小球系膜增生以及肾间质纤维化情况。结果:与模型组比较,治疗6周后,白虎加桂枝汤各剂量组大鼠体质量增加,活动能力、精神状态及毛色好转(P<0.05)。BHJGZ中组可显著降低大鼠血清Scr水平(P<0.01),BHJGZ中组、BHJGZ高组可明显降低大鼠血清BUN水平(P<0.01);白虎加桂枝汤各剂量组均可明显降低HUA大鼠血清UA、24 h UTP水平并改善肾组织形态学、肾小球系膜增生及肾间质纤维。结论:白虎加桂枝汤可能通过抑制肾小球系膜基质增生、减少肾小管损伤及肾间质纤维化,延缓HN进展。
基金supported by the National Natural Science Foundation of China,No.81772327(to HDL)the Shuguang Program of Shanghai Education Development Foundation and Shanghai Municipal Education Commission,China,No.15SG34(to HDL)
文摘Polyethylene glycol can connect the distal and proximal ends of an injured nerve at the cellular level through axonal fusion to avoid Wallerian degeneration of the injured distal nerve and promote peripheral nerve regeneration.However,this method can only prevent Wallerian degeneration in 10% of axons because the cytoskeleton is not repaired in a timely fashion.Reconstruction of the cytoskeletal trunk and microtubule network has been suggested to be the key for improving the efficiency of axonal fusion.As a microtubule-severing protein,spastin has been used to enhance cytoskeletal reconstruction.Therefore,we hypothesized that spastin combined with polyethylene glycol can more effectively promote peripheral nerve regeneration.A total of 120 male Sprague-Dawley rats were randomly divided into sham,suture,polyethylene glycol,and polyethylene glycol + spastin groups.In suture group rats,only traditional nerve anastomosis of the end-to-end suture was performed after transection of the sciatic nerve.In polyethylene glycol and polyethylene glycol + spastin groups,50 μL of polyethylene glycol or 25 μL of polyethylene glycol + 25 μL of spastin,respectively,were injected immediately under the epineurium of the distal suture.Sensory fiber regeneration distance,which was used to assess early nerve regeneration at 1 week after surgery,was shortest in the suture group,followed by polyethylene glycol group and greatest in the polyethylene glycol + spastin group.Behavioral assessment of motor function recovery in rats showed that limb function was restored in polyethylene glycol and polyethylene glycol + spastin groups at 8 weeks after surgery.At 1,2,4 and 8 weeks after surgery,sciatic functional index values and percentages of gastrocnemius muscle wet weight were highest in the sham group,followed by polyethylene glycol + spastin and polyethylene glycol groups,and lowest in the suture group.Masson staining was utilized to assess the morphology of muscle tissue.Morphological changes in skeletal muscle were detectable in sutur
