Combined use of platelet-rich plasma and autologous bone grafts in the treatment of long bone defects in mini-pigs

The use of platelet-rich plasma (PRP) for improving of bone defect healing is discussed controversially. The aim of this study was to assess the effect of PRP in combination with autologous cancellous graft on bone defect healing in a critical metaphyseal long bone defect. A critical size defect in the tibial metaphysis of 16 mini-pigs was filled either with autologous cancellous graft as control group or with autologous cancellous graft combined with autologous PRP. Compared to native blood platelets were enriched about 4.9-fold in the PRP. After 6 weeks, the specimens were assessed by X-ray and histological evaluation. Histomorphometrical analysis revealed that the area of new bone was significantly higher in the PRP group concerning the central area of the defect zone (p < 0.02) as well as the cortical defect zone (p < 0.01). All defects showed substantial new bone formation, but only defects of the PRP group regenerated entirely. The PRP group was superior to the control group even in the semi-quantitative assessment of the osseous bridging in both observed areas of the defect. Within the limits of the present study it could be demonstrated that PRP combined with autologous cancellous graft leads to a significantly better bone regeneration compared to isolated application of autologous cancellous graft in an in vivo critical size defect on load-bearing long bones of mini-pigs.     

Long-term histological results following glutaraldehyde cross-linked collagen injection into the suburothelial space of the mini-pig bladder

Endoscopic correction of vesicoureteric reflux by subureteric injection has become an accepted method of treatment in selected cases. Its efficacy has been demonstrated in the treatment of well over 1,000 ureters. However, there are doubts as to the long-term safety of the commonly injected substance Teflon. Glutaraldehyde cross-linked bovine collagen, a biodegradable substance, was suggested as an alternative material for injection. The aim of this study was to evaluate the histological long-term behavior of glutaraldehyde cross-linked bovine collagen injected into the mini-pig bladder. One hundred and eight deposits of cross-linked collagen, each containing 0.2 ml (n = 36) or 0.6 ml (n = 72) collagen paste, were injected into the suburothelial space of the bladders of 24 mini-pigs. The histological behavior of the implants was studied at monthly intervals over a period of 12 months. After only 2 months a marked invasion of host fibroblasts and formation of endogenous type I and III collagen could be observed. Over 12 months the invasion of fibroblasts and the formation of new collagen increased dramatically. No migration of collagen particles into the local lymphatic system could be observed. A new staining technique (solophenyl red 3BL) was introduced to selectively demonstrate type I and III collagen fibres. In addition, the efficacy of the subureteric collagen injections could be demonstrated by successfully treating reflux that was iatrogenically induced in the mini-pig bladders.     

一次性静脉注射高剂量链脲佐菌素建立1型糖尿病小型猪模型

目的 探讨通过一次性注射高剂量链脲佐菌素（streptozotocin,STZ）方法建立1型糖尿病小型猪模型的可行性。方法 中华实验小型猪耳缘静脉一次性注射链脲佐菌素溶液150 mg/kg,分别在给药前和给药后10 min、30 min、90 min、第1天、第2天、第3天和第7天空腹采集静脉血,动态监测空腹血糖,并利用静脉糖耐量实验和C肽释放实验对模型进行鉴定。结果 给药后第1天开始,模型组空腹血糖明显升高并始终维持在16.7～20.6 mmol/L的浓度范围,达到糖尿病标准;静脉葡萄糖耐量试验和C肽释放实验结果表明,静脉注射体积分数50%的葡萄糖1 h后模型猪血糖浓度高于11.1 mmol/L,2 h后未能恢复至空腹血糖水平;而胰岛素和C肽在注入葡萄糖后基本未发生任何反应,始终保持痕量水平。结论 一次性静脉注射大剂量链脲佐菌素的方法能够成功建立1型糖尿病小型猪模型。     

小型猪肝硬化模型肝脏病理及生化指标的变化

目的 探讨小型猪肝硬化模型制备过程中肝脏病理学及生化指标的变化。方法 选用同种系巴马小型猪50头,随机分为实验组40头,对照组10头,雌雄不限,4月龄,体重15-20kg。实验组予四氯化碳复合因素造模法喂养,正常组予饲料及清水喂养。于造模的0、4、8、12、16、20、24周末进行肝脏穿刺活检及耳缘静脉抽血进行数据采集,对比分析肝纤维化-肝硬化制模过程中的肝脏病理学及生化指标的变化。结果24周末,对照组获得数据S0期70例次;实验组获得数据轻度肝纤维化（S1＋S2期）82例次,重度肝纤维化（S3＋S4期）62例次,肝硬化共39例次。从正常组到肝硬化组谷丙转氨酶、谷草转氨酶、总胆红素、血清肌酐及球蛋白呈逐渐升高的趋势,而白蛋白,A/G比值呈逐渐下降的趋势,凝血酶原时间则逐渐延长,统计学分析,组间存在明显差异（P＜0.01）。结论 四氯化碳复合因素造模法可以成功制备出小型猪肝硬化模型,肝纤维化不同阶段肝脏病理学和生化指标均发生明显变化。     

2型糖尿病小型猪勃起功能障碍及NOS变化

目的观察一氧化氮合酶(NOS)在糖尿病性勃起功能障碍中的作用。方法采用高脂高糖饮食喂养的方法建立广西巴马小型猪糖尿病性阴茎勃起功能障碍模型(DMED),对正常对照组(CD)和高脂高糖组(HFSD)动物于造模第12月后进行阿朴吗啡阴茎勃起实验,用酶联免疫吸附试验(ELISA)检测阴茎海绵体组织中NOS活性。结果 HFSD组动物空腹血糖和血清总胆固醇明显升高(P0.01),HFSD组动物阴茎勃起率较正常组明显降低(P0.01),NOS活性亦显著降低(P0.05)。结论糖尿病严重影响小型猪勃起功能,阴茎海绵体组织NOS活性的降低可能是其机理之一。     

一种改进的Guyton流导测定法及对动物经脉流导的测量

在Ｇｕｔｏｎ１９６６年创立的流导测定法基础上，我们创立了一种新方法。用压差传感器代替Ｇｕｙｔｏｎ实验中的弹性腔和压力传感器产生压力差并直接转换成电信号输入计算机，使用两个ＷＩＮ针头作测量，通过对数转换和线性回归计算出流导率。在麻醉的大白鼠和小型猪活体的循经低电阻线皮下组织和对照区上进行了测量，分别使用２．５ｃｍＨ２Ｏ和１０ｃｍＨ２Ｏ两种压力差。结果表明，大白鼠经脉和非经脉组织的流导率分别为１４．５±１５．９２和９．５５±１４，０１（×１０－８，ｃｍ４／ｄｙｎｅ－ｓｅｃ），小型猪经脉组织和非经脉组织的流导率分别为１，７２±２．１９和０．７８±３．１７（×１０－５；ｃｍ４／ｄｙｎｅ－ｓｅｃ），经脉上的流导率比经脉外显著高（Ｐ＜０．０５）。本方法可消除同步干扰，两种压力差下测得的流导率数值与Ｇｕｙｔｏｎ在非水肿和水肿两种状态下测得的流导率基本相同，说明了间隙组织流导的生物流变特性。     

西藏小型猪MSTN基因分子克隆及序列分析

目的扩增西藏小型猪的肌肉生长抑制素(MSTN)基因编码区全长序列,并对序列进行生物信息学分析。方法提取西藏小型猪肝脏组织的RNA,反转录成为c DNA,利用RT-PCR方法扩增MSTN基因编码区全长序列,将纯化的片段与pMD19-T载体连接,转化大肠杆菌DH5α,筛选阳性克隆并测定其序列,利用生物信息学方法分析其序列特征,并与其他12种物种构建系统进化树。结果西藏小型猪MSTN基因编码区全长1128bp,编码375个氨基酸,氨基酸序列同源性分析表明,与版纳微型猪MSTN氨基酸序列相似性最高,为99%。结论 MSTN基因西藏小型猪除了与鸡和斑马鱼亲缘关系较远外,与人、犬、版纳微型猪、绵羊、山羊、牛、马、黑猩猩、大鼠、小鼠的关系都较为接近,其中与版纳微型猪的亲缘关系最为接近。     

猪体外并行循环模型的建立及管理

目的 探讨小型猪体外并行循环模型的建立及管理.方法 小型猪12例,全麻诱导插管后,开放股动、静脉建立体外循环,并于建立体外循环前与建立体外循环后每1h监测动脉压、心率、血氧饱和度、呼气末二氧化碳分压、中心静脉压以及红细胞比积.结果 体外循环建立前后监测数据均在正常范围内,两者在统计学上无明显差异(P＞0.05).结论 小型猪并行体外循环在实验中具有稳定性,可作为CPB相关研究的动物模型.     

24h内不同俯脚卧位通气时间比对急性肺损伤模型病理的影响

目的比较24h内不同俯“印卧位辅助通气时间比对小型猪急性肺损伤（ALI）病理结果的影响。方法选取7曲个月龄健康雌性小型猪15只，平均体质量（30．2-3．2）kg，身长（76．5±5．9）cm。麻醉后行气管插管机械通气，经气管插管注射pH1．0盐酸制备呼吸窘迫综合征（ARDS）模型。随机分为8h组（俯卧8h±仰卧16h组），16h组（俯卧16h±仰卧8h组），24h组（持续俯卧24h组）各5只小型猪。24h后对模型动物实施安乐死，取出肺组织观察大体标本，计算肺湿干重比（W，D）。肺组织甲醛固定，苏木素-伊红（HE）染色切片，参照半定量评分标准进行评分。结果3组间肺炎评分与W／D比较，16h组与24h组均低于8h组，但差异不显著。结论早期足量（16h或以上）的俯卧位辅助通气可能减少急性肺损伤动物模型的肺部炎性反应。