高寒环境实验兔骨骼肌火器伤组织病理学观察

目的:探讨高寒低温干燥自然条件下,火器伤病理组织学变化特点,为临床救治及预防冻-火器复合伤提供依据。方法:对一组实验兔,在-18℃～-22℃低温干燥自然条件下,用5.62 mm小口径手枪,距0.5cm射击双后肢,制做冻-火器复合伤模型;观察骨骼肌火器伤病理组织学改变及超微结构特征。结果:高寒低温干燥自然条件下冻-火器复合伤的病理改变,挫伤区组织以坏死为主,震动区以变性为主。提示高寒低温环境肌组织火器伤病理改变是火器与高寒低温共同引起损伤,高寒条件加剧损伤程度;而在创伤肌组织震荡区中,仍保存较多的肌性修复功能,呈现以肌巨细胞增生性为主的修复过程。结论:高寒干燥自然条件下早期脱离高寒环境,可以较好地减轻或预防冻-火器复合伤的程度。     

猪腹部肠管火器伤后肠源性内毒素血症与胃损伤

目的:探讨腹部火器伤肠管穿透后胃的损伤性变化.方法:健康长白仔猪42头随机等分为对照组和腹部火器伤肠管穿透伤后1h、2h、4h、8h、12h和24h实验组.测定各组动物血浆内毒素,光镜下观察各组胃的组织形态学变化.结果:实验组各组血浆内毒素水平均明显高于对照组,内毒素于伤后8小时出现高峰,伤后12小时仍维持在高峰值水平(p<0.05);伤后各组出现逐渐加重的胃粘膜下充血、水肿,12h、24h组出现炎细胞浸润、浆膜下出血.结论:腹部火器伤肠管穿透后,胃损伤性变化随时间延长而加重;肠源性内毒素血症在火器伤后继发性胃损伤中可能起重要作用.     

急性兔脑栓塞模型的建立

目的建立稳定的兔脑栓塞模型,以期为进一步开展脑栓塞的病理生理变化及影像学研究提供可靠实用的工具。方法30只健康新西兰兔,随机分成3组,其中A组3只,为空白对照组;B组5只,为假手术对照组;C组22只,为栓塞组。分离右侧颈部血管,经颈外动脉向颈内动脉注入直径约0.5~1.0 mm的SiO2颗粒10枚左右,栓塞后30 min行CT灌注检查,利用多层螺旋CT灌注成像对各组动物的脑缺血情况进行观察。24 h处死动物取脑组织进行病理研究。结果A、B组CT灌注及病理均未见异常。C组栓塞过程中3只兔死亡。16只兔CT灌注异常,表现为右侧局部CBF降低、MTT延长、CBV无明显变化或轻度上升、下降。3只兔灌注未见异常。HE染色可见8只兔脑梗塞,7只兔脑缺血,4只兔未见明显异常。结论采用该方法和技术能够建立稳定的兔脑栓塞模型,结果可靠,具有可操作性和重复性。     

兔体外辅助循环模型的建立

用40只新西兰兔,在常温不停跳下,采用左心房插管引流,左股动脉灌注的方法,建立兔左心辅助体外循环动物模型,39只模型成功建立.术中血压、心率及体温等各项生理指标基本稳定,与开胸前柑比,各项指标变化均小于5%.     

兔不同急性肝衰竭模型的建立与评价

目的:探讨一种理想的急性肝衰竭(AHF)模型建立方法.方法:36只实验兔随机分为3组:①改良药物手术诱导组(A组,n=12),先用D-氨基半乳糖(D-Gain)和脂多糖(LPS)腹腔注射,同时加用乳果糖,注射后2h以氟烷作为麻醉剂,切除约50%肝脏组织.术中经肝静脉注入5%葡萄糖氯化钠溶液10ml/kg体重;②传统手术诱导组(B组,n=12),切除约95%肝脏组织,术中不行肝静脉穿刺注射5%葡萄糖氯化钠溶液;③药物诱导组(C组,n=12),用D一氨基半乳糖(D-Gain)和脂多糖(LPS)一次性腹腔注射.比较建模死亡率、建模后24h兔存活率、血谷丙转氨酶(ALT)、血氨(NH3)、总胆红素(TB)和血糖(BS).结果:B组手术死亡率高于A组死亡率(41.77%vs0%),A组、B组兔建模成功后24h存活率及C组兔建模成功后72h存活率分别为0%,0%,25%,A组ALT和NH3水平显著高于C组(P＜0.05),TB和BG水平低于C组,但差异无显著性.结论:通过改进的50%肝切除术可建立较理想的兔AHF模型,以氟烷作为麻醉剂,药物诱导注射同时加用乳果糖,术中经中叶肝静脉注入5%葡萄糖氯化钠溶液可减少手术死亡率.     

兔脓毒性休克模型的建立

目的:建立相对稳定的兔脓毒性休克模型.方法:雄性新西兰大白兔20只,随机分为模型组和假手术组.麻醉后无菌操作下取中、下腹部正中切口,找出盲肠,距盲肠末端8.0cm处4号丝线结扎盲肠和相应血管;于盲肠游离末端避开血管戳孔2次,孔径0.5cm;将盲肠原位放回腹腔,缝合腹壁切口;腹腔内注射30ml/kg温生理盐水.假手术组只探查腹腔.术后每隔3小时监测肛温.颈动脉插管有创动脉压监测确定模型成功后送血培养、血气、血乳酸,监测4小时后处死动物,送腹水培养,取右肺中叶测定肺含水量,取心、肝、脾、肺、肾、肠常规切片HE染色.结果:制模成功时间为:(18.91±1.384)小时;模型平均动脉压MAP(95.00±10.817 vs 52.38±15.565,P＜0.05);血气:PH(7.40±0.047 vs 7.09±0.146,P＜0.01),PCO2(30.0±5.831 vs 19.80±4.104,P＜0.01),BE(-6.46±4.931 vs -24.11±4.276,P＜0.01),HCO3-(18.45±4.367 vs 5.73±2.422,P＜0.01);血乳酸(2.53±1.108 vs 7.85±5.834,P＜0.05);血培养(7/10)阳性:大肠埃希菌;腹水培养(10/10)阳性:大肠埃希菌及阴沟肠杆菌;肛温于术后呈先上升后下降,假手术组肛温较模型组差异有显著性(39.63±0.492 vs 36.82±0.999,P＜0.01);成模后肛温与平均动脉压呈正相关关系,r=0.748.P=0.013,方程:y(平均动脉压)=34.46x+0.45;肺干湿比及肺含水量无明显差异[(0.22±0.014 vs 0.19±0.288,P＞0.05),(78.17±1.375 vs 80.58±2.878,P＞0.05)].常规HE染色可见明显病理学改变.结论:本模型可模拟多微生物感染致脓毒性休克模型,模型相对稳定,可用于兔种属.     

兔慢性肾功能衰竭模型的建立与评价

目的建立兔慢性肾功能衰竭模型,为干细胞移植治疗和相关研究奠定基础。方法普通级大耳白兔随机分为正常对照组和单侧输尿管结扎（unilateral ureteral obstruction,UUO）组。UUO组于输尿管结扎后2、4、6、8周进行血生化肾功能指标检测,并取肾组织观察肾脏病理学改变,通过SPECT动态观察肾小球滤过率的变化,采用免疫组织化学方法观察肾组织转化生长因子-β1（TGF-β1）的表达情况。结果①UUO组术后第2周,出现明显的血肌酐升高,尿素氮术后第8周开始升高（P〈0.01）。②UUO组术后第4周,肾脏组织出现了早期间质纤维化的病理改变,术后第8周肾小球开始出现硬化,间质纤维化明显,皮质明显变薄。术后第12周,肾小球硬化比例增加,肾小管玻璃样变性,间质纤维化进一步加重（P〈0.05）。③SPECT动态观察肾小球滤过率,UUO组第4周GFR值比正常对照组降低,到第8周时,GFR值进一步下降,结扎侧肾脏功能降低甚至丧失。④免疫组织化学染色显示,TGF-β1在术后第4、8、12周均明显增强,并且各时间点表达均有显著差异（P〈0.05）。结论单侧输尿管结扎法成功制作比较稳定的慢性肾功能不全模型,UUO后第8周符合肾脏间质纤维化模型标准。     

猪腹部火器伤肠管穿透后心肌损伤的变化

目的:观察腹部火器伤肠管穿透后心肌损伤的变化.方法:健康长白仔猪42头随机分为对照组以及伤后1h、2h、4h、8h、12h和24h组.实验组建立腹部火器伤肠管穿透模型后,分别测定伤后1 h、2h、4h、8h、12h和24h组各组动物血清中LDH、CK、CK-MB水平,并与对照组比较,观察实验组各时间点心脏组织学变化?结果:伤后各组血清LDH、CK、CK-MB水平均高于对照组.伤后8h、12h、24h组光镜下出现逐渐加重的心肌细胞水肿、变性;电镜下4h、8h、12h、24h组出现逐渐的线粒体肿胀、溶解;对照组光、电镜下未见明显的损伤性变化.结论:腹部火器伤肠管穿透导致心肌形态和酶的损伤性变化,随着伤后时间的延长而加重.     

猪腹部火器伤肠管穿透后继发性脑损伤变化

目的:观察腹部火器伤肠管穿透后继发性脑损伤的变化.方法:健康长白仔猪42头随机分为对照组以及伤后1h、2h、4h、8h、12h和24h组,实验组建立腹部火器伤肠管穿透模型后,观察实验组各时间点脑组织学和超微结构的变化.结果:伤后12h、24h组光镜下出现逐渐加重的神经细胞水肿、变性;电镜下8h、12h、24h组出现逐渐加重的内质网扩张、线粒体肿胀、溶解;对照组光、电镜下未见明显的损伤性变化.结论:腹部火器伤肠管穿透后导致继发性脑组织形态学变化,随着伤后时间的延长而逐渐加重.     

火器伤合并海水浸泡后对大鼠伤口愈合的影响

目的制造火器伤合并海水浸泡后的动物愈合模型,观察火器伤合并海水浸泡后对伤口愈合过程的影响。方法20只雄性Wistar大鼠随机分为2组：对照组（n=10）及实验组（n=10）。对照组为单纯火器伤。实验组（海水浸泡组）为单纯火器伤后合并海水浸泡30 min。检测愈合过程中2组动物体重增加量变化、面积变化,并对已愈伤口组织行病理学检查。结果实验组伤口愈合时间为16-20 d（17.7±1.3）d,对照组为14-17 d（15.8±0.6）d。对照组动物体重平均增加量显著高于实验组。实验组已愈合伤口组织中存在特殊的病理学变化。     

百草枯致急性肺损伤大鼠模型的建立

目的建立一种百草枯诱导的急性肺损伤（ALI）大鼠模型。方法将20只SD大鼠随机分为正常对照组10只、实验组10只。实验组一次性口服灌胃百草枯（PQ）80mg／kg,于给药后1天处死大鼠,观察光镜下肺组织病理改变、肺动脉血氧分压（PaO2）、支气管肺泡灌洗液（BALF）蛋白含量等。结果给予百草枯1天后肺形态学出现显著异常,PaO2及BALF蛋白含量出现显著改变。结论一次性灌胃百草枯80mg／kg成功建立急性肺损伤动物模型。     

N-Terminal Arginylation of Sciatic Nerve and Brain Proteins Following Injury

N-terminal protein arginylation has been demonstrated in vitro and in situ and has been reported to increase following injury to sciatic nerves of rats. The present study attempts to demonstrate these reactions in vivo by applying [³H]Arg to the cut end of sciatic nerves in anesthetized rats and assaying for N-terminal arginylation using Edman chemistry and acid precipitation of labeled proteins in the proximal nerve segment. No evidence was found for arginylation in an aqueous soluble fraction. However, N-terminal arginylation was detected in a urea soluble fraction at 2 hours after nerve crush. The data show that arginylation of rat sciatic nerve proteins occurs in vivo and suggest that the arginylated proteins formed an aqueous insoluble/urea soluble aggregate after arginylation. In other experiments, rat brains were injured and assayed for arginylation in vitro to test the hypothesis that injury causes an up-regulation of these reactions. Results showed an activation of the reaction at 2 hours post crush and indicate that increases in N-terminal arginylation are likely to be a general response to injury in nervous tissue.     

Spatiotemporal Expression of PSD-95 and nNOS After Rat Sciatic Nerve Injury

Neuronal nitric oxide synthase (nNOS) has been implicated to influence peripheral nerve lesion and regeneration. Post-synaptic density-95 (PSD-95) is one of nNOS-anchoring proteins and plays an important role in specifying the sites of reaction of NO in nervous system. Here we established a rat sciatic nerve crush (SNC) model to examine the spatiotemporal expression of PSD-95 and nNOS. At gene levels, PSD-95 mRNA diminished shortly after crush, and significantly elevated from 2 days to 2 weeks, whereas nNOS decreased progressively post-operation, reached the valley at 1 day, and markedly up-regulated from 1 to 2 weeks after SNC. The expression of both molecules returned to the control level at 4 weeks post-injury. At protein levels, PSD-95 and nNOS underwent the similar changes as their gene expression except for a time lag during up-regulating. At their peak expression, PSD-95 co-labeled with nNOS in Schwann cells (SCs) of sciatic nerve within 0.5 mm from the lesion site, but had few colocalization in axons. In addition, the interaction between PSD-95 and nNOS enhanced significantly at 2 weeks after SNC. These results suggest a correlation of PSD-95 up-regulation with nNOS in reactive SCs of crushed sciatic nerve, which may lead to understanding the function of PSD-95 during peripheral nerve regeneration. Shangfeng Gao and Min Fei contributed equally to this work.     

阿霉素心肌损伤大鼠模型的制备与评价

目的制备阿霉素心肌损伤大鼠模型,并对其进行评价。方法24只雄性SD大鼠随机分2组：正常对照组（CON,n=9）和阿霉素模型组（ADR,n=15）。ADR组腹腔注射阿霉素2 mg/kg,每周3次,连续2周,CON组注射相同体积的生理盐水,注射完毕后饲养5周;实验期间观察大鼠一般情况及死亡率;7周后检测心脏血流动力学及组织形态学变化,并进行心肌氧化损伤生化测定。结果ADR组大鼠死亡率为40%,CON组无死亡。与CON组比较,ADR组大鼠左室舒张末压（LVEDP）及左室内压最大下降速率（-LVdP/dtmax）显著升高（P〈0.001,P〈0.05）;组织学检查结果符合心肌损伤病理学改变的典型特征;心肌丙二醛（MDA）含量明显增加（P〈0.001）;谷胱甘肽过氧化物酶（GSH-Px）活性显著降低（P〈0.01）。结论按12 mg/kg的ADR总剂量,以每周3次,共两周,每次2mg/kg腹腔注射方式给药,7周后大鼠心脏产生明显功能及形态学异常,可成功建立ADR心肌损伤大鼠模型。     

[PEAD:肝素:NGF]生物材料促进大鼠坐骨神经损伤恢复

目的:探讨新型材料poly(ethylene argininylaspartate diglyceride)(PEAD)结合肝素包裹神经生长因子组成的三元复合体比单纯运用NGF治疗大鼠坐骨神经损伤效果明显,为临床治疗外周神经损伤提供实验依据。方法:24只200g左右Wistar大鼠,分成生理盐水组,NGF组,NGF凝聚体三组,每组各8只,距梨状肌下缘远侧约1.5cm处运用静脉夹夹紧坐骨神经2min,采用无创细线(5/0)缝合肌肉和皮肤,并用碘伏进行消毒,NGF组每天沿坐骨切迹肌注80ngNGF,持续30天;NGF凝聚体组仅在造模时肌注复合体(内含2.4μg的NGF);生理盐水组给予等体积的生理盐水。术后每周运用脚步印迹法评价动物的行为学,并于30天后灌流、收集各组损伤侧坐骨神经,运用HE染色及投射电镜观察坐骨神经结构恢复情况,免疫荧光标记MBP,观察其蛋白的表达。结果:NGF组,NGF凝聚体组在行为学、病理结构及蛋白的表达远高于生理盐水组,并且NGF凝聚组的治疗效果优于NGF组。结论:新型凝聚体包载NGF具有明显的促进周围神经损伤后的修复与再生作用,能够在一定程度上提高单纯运用NGF治疗大鼠坐骨神经损伤的不足,达到更加理想和显著的促恢复效果。     

家兔子宫内膜炎模型的建立和临床病理学观察

目的建立家兔子宫内膜炎模型,观察模型的临床病理学变化,为家畜子宫内膜炎的诊断和防治研究提供材料。方法造模通过子宫灌注病原菌的方法,临床病理学观察通过临床症状、体温、血常规、子宫分泌物和尿液检查、子宫剖检、子宫内膜的显微和超微结构观察。结果模型兔精神变差、采食量减少,阴门肿胀,流出脓性分泌物;每次灌注细菌后8 h内体温升高0.5℃左右,血液中性粒细胞的比例升高;子宫分泌物中混有大量白细胞、脓球和少量脱落的子宫内膜上皮细胞;子宫显著肿大,子宫内膜溃疡和出血,上皮结构不完整,细胞变性、坏死,微绒毛和纤毛脱落,上皮下出血、瘀血、水肿和炎性细胞浸润。结论通过子宫灌注病原菌的方法可以成功制备家兔子宫内膜炎模型,模型的临床病理变化以局部炎性为主。     

Investigations on the Incorporation of a Tritiated Ganglioside, GM1, in the Injured and Intact Sciatic Nerves of the Rat

Following injury of their left sciatic nerves by means of a standardized procedure, male rats received intravenous injections of a tritiated ganglioside. GM1, on different days during the process of regeneration. The rats were killed at two different times after the injection and the concentrations of the total radioactivity, nonvolatile radioactivity, and labelled GM1 were estimated in six segments of the crushed and intact sciatic nerves. The segments of the damaged nerves showed higher concentrations of radioactivity and a higher content of GM1 than the corresponding segments of the contralateral nerves. Within the immediate area of the lesion the highest levels were found on the 3rd and 6th days after the injury; the segments distal from the lesion showed the highest levels of activity on days 9 and 12. The nerve segments proximal to the site of the injury showed a low rate of radioactivity incorporation. The higher concentrations of [3H]GM1 in damaged nerves as well as the rate of incorporation as a function of time indicate that exogenous gangliosides may be involved in the processes of regeneration and have a bearing on the latter.     

Changes in Aldose Reductase After Crush Injury of Normal Rat Sciatic Nerve

The response of aldose reductase (AR) to crush injury was studied in normal rat sciatic nerve. Enzyme activity and immunoreactivity of AR were determined at intervals of 1, 5, 14, 28, and 35 days after crush and correlated with histologic and immunocytochemical observations. During nerve degeneration in the distal segments of crushed nerves, a significant reduction in AR activity was detected. At 5 and 14 days, coincident with Schwann cell proliferation, enzyme activity decreased by nearly two- and fourfold, respectively. Although activity of AR increased by 28 days during nerve regeneration, it was not restored to normal levels at 35 days. Similar reductions were observed with the immunoblotting of the enzyme. Quantitative analysis of immunogold labelling on electron micrographs confirmed that proliferating as well as remyelinating Schwann cells contained reduced gold particle density compared to Schwann cells of noncrushed myelinated fibers. Immunoblots of P0, a marker for the degree of Schwann cell differentiation or myelination, showed that the temporal sequence of changes in P0 paralleled that of AR. Thus expression of AR is a function of differentiated or mature Schwann cells. The putative volume regulatory role of AR in Schwann cells may become superfluous during Wallerian degeneration.     

Axonal Transport Characteristics of Gangliosides in Sensory Axons of Rat Sciatic Nerve

The distribution of axonally transported gangliosides and glycoproteins along the sciatic nerve was examined from 3 h to 4 weeks following injection of[3H]glucosamine into the fifth lumbar dorsal root ganglion of adult rats. Incorporation of labeled precursor into these glycoconjugates reached a maximal level in the ganglion within 6 h. Outflow patterns of radioactivity for glycoproteins showed a well-defined crest with a transport rate of approximately 330 mm/day. In contrast, the crest of transported gangliosides was continuously attenuated, implying a significant deposition along the axon, and an alternative method of calculating velocity was required. Analysis of accumulation of labeled material at double ligatures demonstrated both anterograde and retrograde transport of glycoproteins and gangliosides and allowed for the calculation of an anterograde transport rate of about 270 mm/day for each. Additional evidence of ganglioside transport is provided in that the TLC pattern of transported radioactive gangliosides accumulating at a ligature is significantly different from the pattern seen in the dorsal root ganglion or following intraneural administration of the labeled precursor. These data indicate that gangliosides are transported at the same rapid rate as glycoproteins but are subject to a more extensive exchange with stationary material than are glycoproteins.