丁香配伍郁金对胃肠运动影响的药理实验研究(Ⅲ)——丁香配伍郁金对病理模型的影响

目的 :研究丁香配伍郁金后不同有效部分对模型动物胃肠运动的影响。方法 :用阿托品或吗丁啉模拟胃肠运动抑制或亢进 ,采用水蒸气蒸馏法提取挥发油 ,观察丁香配伍郁金后不同有效部分对小鼠胃排空及小鼠印度墨汁推进实验的影响。结果 :丁香煎液的药理作用可被郁金煎液减弱 ,呈现丁香恶郁金的关系。结论

心肌缺血动物实验技术方法在机能实验教学中的创新

心肌缺血动物模型的制备方法很多,目前应用比较广泛的是结扎左冠状动脉前降支复制心肌缺血模型[1]。根据多年科研与教学实验,发现如果实验动物外科手术顺利,术后抗感染和术后营养达到标准,给药组大鼠实际死亡量很小,而且各种生理功能指标均能恢复,实验动物甚至日常活动跟正常对照组,假手术组一样。但是同样的药物治疗对于人类心肌缺血患者尤其是长期慢性患者来说,治疗的效果不佳,有些人不得不接受支架,搭桥手术。

络脉绌急模型大鼠炎症相关细胞因子的变化研究

目的:探讨络脉绌急模型大鼠血清中TNF-α、IL-2、IL-6和IL-10等炎症相关细胞因子的变化规律,为阐明络脉绌急病理变化奠定理论基础。方法:模型组大鼠每天灌服L-蛋氨酸(1 g/kg),分别于第10 d、第20 d、第30 d腹腔注射垂体后叶素(10 U/kg),正常组大鼠则注射生理盐水。注射后1 h左颈总动脉采血,室温凝固后于4℃,2000转/min,离心15m in,分离血清检测TNF-α、IL-2、IL-6和IL-10的含量;结果:造模10 d时,模型组大鼠血清的TNF-α、IL-2、IL-6水平均升高,并且20 d、30 d时继续升高,与正常对照组比较差异有统计学意义,而IL-10水平在开始时升高(造模10 d、20 d),随后下降。结论:利用喂食蛋氨酸损伤血管内皮细胞,复制络脉绌急大鼠模型时可能引发炎症反应病理过程。

神经源性肌萎缩病理动物模型的制备

神经源性肌萎缩常见的原因为废用、营养障碍、缺血和中毒。脊髓前角病变、神经根、神经丛、周围神经的病变等均可引起神经兴奋冲动的传导障碍，从而使部分肌纤维废用，产生废用性肌萎缩。常见的疾病有：脊椎椎骨骨质增生、椎间盘病变、脊神经肿瘤、蛛网膜炎、神经炎、神经丛病变、脊神经肿瘤、神经损伤、脊髓空洞症、运动神经元性疾病、格林一巴利综合征、脑部病变和脊髓病变等。本文试用大鼠坐骨神经双重结扎的方法复制神经源性肌萎缩动物模型。

药物制剂靶向性评价方法的研究进展

靶向给药系统（targeting dmg delivery system,TDDS）又称靶向制剂,是通过载体使药物选择性的浓集于病变部位的给药系统。病变部位常被称为靶部位,可以是靶组织、靶器官,也可以是靶细胞或细胞内的某靶点。靶向给药可以增加药物在靶部位的浓度、降低药物在非靶部位的浓发、延长药物在靶部位的停留时间,从而提高给药后的疗效。药物制剂靶向系统需要建立一个能够全面、客观、准确地反映靶向性的评价体系,一方面可对制剂的靶向性有一个可视化的数据或图、表支持;另一方面可为制剂进行条件优化提供实验依据,因此为靶向性建立一个好的评价体系是在研究靶向制剂过程中必不可少的。靶向制剂的研制需要有可靠的评价标准来约束,这就刺激了各种评价标准的研究,建立好的评价标准,可以指导靶向制剂的研制,使其能够更好的满足临床的需要。本文就目前在药物靶向制剂领域应用比较广泛的靶向性评价方法进行综述。

慢性咽炎血液流变学实验观察

目的 探讨慢性咽炎的发生机理。方法 采用氨水和松节油复合因素在日本大耳白兔咽部直接塑造慢性咽炎动物病理模型 ,设立正常对照组、病理模型组测动物血液流变学指标。结果 显示造模 2周时 ,模型组兔全血低切粘度、血浆粘度、红细胞刚性指数、红细胞聚集指数、血浆纤维蛋白含量较正常对照组显著增高 (P <0 .0 1)。结论 提示慢性咽炎可能有血液流变学的异常。

大鼠局灶性脑缺血白细胞流变性变化及乌龙丹的作用

目的 通过建立大鼠局灶性脑缺血病理模型,观察其白细胞流变性的变化,并探讨活血化瘀中药复方乌龙丹的影响。方法 选用Wistar大鼠结扎颈总动脉(CCA)后再阻断大脑中动脉(MCA)造模,实验分为假手术、模型、乌龙丹低剂量(WLDL)、乌龙丹高剂量(WLDH)四组,造模术后24h取血测定白细胞总数、白细胞变形指数、白细胞滤过堵塞的粒子浓度及红、白细胞滤过指数IF-红细胞滤过指数IF。结果 模型组的白细胞总数、白细胞变形指数、白细胞滤过堵塞的粒子浓度及红、白细胞IF-红细胞IF均明显高于对照组(P<0.01),表示白细胞的变形能力下降。乌龙丹给药组上述指标与模型组有显著性差异(P<0.01),显示出该方对梗塞后白细胞变形能力下降有一定的对抗作用。结论 急性局灶性脑缺血损伤中有白细胞流变性异常,变形能力下降。中药复方“乌龙丹”对此有一定对抗作用,增强白细胞的流变性和使其黏附性降低,是其活血化瘀作用的重要方面。

Anticancer Effect of PS-T on the Experimental Hepatocellular Carcinoma

Objective: To apply PS-T in di?erent phases of carcinoma formation and development, and research the mechanism of anti-carcinoma of PS-T in the cytological level. Methods: N-nitrosodiethylamine (DENA) and CCl4 were applied jointly to duplicate the rat hepatocirrhosis and hepatic cancer model. The rats were divided into 7 groups and were administrated via nasal-stomach tube with PS-T in di?erent phases to interfere the cancer genesis and development. All the rats were killed in 20 weeks for pathological observation. Results: The loss of body weight of rats slowed down in the PS-T-treated group, and the carcinogenesis rate was signi?cantly decreased correspondingly. PS-T could also inhibit the carcinogenesis by supressing the hepatocirrhosis, which showed the positive correlation between the curative e?ect and the curative period. Conclusion: Application of PS-T during cancer induction showed a signi?cant e?ect on preventing and supressing cancer. PS-T might be an ideal drug for clinical anti-cancer therapy. And it will be a main drug in both combined and single treatments for tumor.

Correlation between TIMP-1 expression and liver fibrosis in two rat liver fibrosis models

AIM： To evaluate serum TIMP-1 level and the correlation between TIMP-1 expression and liver fibrosis in immuneinduced and CCL4-induced liver fibrosis models in rats. METHODS： Immune-induced and CCL4-induced liver fibrosis models were established by dexamethasone （0.01 mg） and CCL4 respectively. Serum TIMP-1 level was detected with ELISA, while histopathological grade of liver biopsy was evaluated. Spearman rankcorrelation test was used to analyse the difference of the correlation between the TIMP-1 expression and hepatic fibrosis in the two fibrosis models. Furthermore,in situ hybridization was used to determine the expression difference of TIMP-1 mRNA in the two models. RESULTS： Positive correlation existed between serum TIMP-1 level of immune induced group and the histopathological stages of fibrosis liver of corresponding rats （Spearman rank-correlation test, rs = 0.812, P 0.05）, and the positive in situ hybridization signal of TIMP-1 mRNA was strong. In CCL4-induced liver fibrosis model, the correlation between the serum TIMP-1 level and the severity of hepatic fibrosis was not statistically significant（Spearman rank-correlation test, rs = 0.229, P 〉 0.05）. And compared with immune-induced model, the positivein situ hybridization signal of TIMP-1 mRNA was weaker, while the expression variation was higher in hepatic fibrosis of the same severity. CONCLUSION： The correlations between TIMP-1 expression and liver fibrosis in two rat liver fibrosis models are different. In immune-induced model, serum TIMP-1 level could reflect the severity of liver fibrosis, while in CCL4-induced model, the correlation between the serum TIMP-1 level and the severity of hepatic fibrosis was not statistically significant.

New animal models for hepatitis C viral infection and pathogenesis studies

Hepatitis C virus （HCV） is a major cause of chronic liver disease, cirrhosis and hepatocellular carcinoma （HCC）. In man, the pathobiological changes associated with HCV infection have been attributed to both the immune system and direct viral cytopathic effects. Until now, the lack of simple culture systems to infect and propagate the virus has hampered progress in understanding the viral life cycle and pathogenesis of HCV infection, including the molecular mechanisms implicated in HCV-induced HCC. This clearly demonstrates the need to develop small animal models for the study of HCV-associated pathogenesis. This review describes and discusses the development of new HCV animal models to study viral infection and investigate the direct effects of viral protein expression on liver disease.

Animal models of portal hypertension

Animal models have allowed detailed study of hemodynamic alterations typical of portal hypertension and the molecular mechanisms involved in abnormalities in splanchnic and systemic circulation associated with this syndrome. Models of prehepatic portal hypertension can be used to study alterations in the splanchnic circulation and the pathophysiology of the hyperdynamic circulation. Models of cirrhosis allow study of the alterations in intrahepatic microcirculation that lead to increased resistance to portal flow. This review summarizes the currently available literature on animal models of portal hypertension and analyzes their relative utility. The criteria for choosing a particular model, depending on the specific objectives of the study, are also discussed.

Basal physiological parameters in domesticated tree shrews (Tupaia belangeri chinensis)

Establishing non-human primate models of human diseases is an efficient way to narrow the large gap between basic studies and translational medicine. Multifold advantages such as simplicity of breeding, low cost of feeding and facility of operating make the tree shrew an ideal non-human primate model proxy. Additional features like vulnerability to stress and spontaneous diabetic characteristics also indicate that the tree shrew could be a potential new animal model of human diseases. However, basal physiological indexes of tree shrew, especially those related to human disease, have not been systematically reported. Accordingly, we established important basal physiological indexes of domesticated tree shrews including several factors: (1) body weight, (2) core body temperature and rhythm, (3) diet metabolism, (4) locomotor rhythm, (5) electroencephalogram, (6) glycometabolism and (7) serum and urinary hormone level and urinary cortisol rhythm. We compared the physiological parameters of domesticated tree shrew with that of rats and macaques. Results showed that (a) the core body temperature of the tree shrew was 39.59±0.05 °C, which was higher than that of rats and macaques; (b) Compared with wild tree shrews, with two activity peaks, domesticated tree shrews had only one activity peak from 17:30 to 19:30; (c) Compared with rats, tree shrews had poor carbohydrate metabolism ability; and (d) Urinary cortisol rhythm indicated there were two peaks at 8:00 and 17:00 in domesticated tree shrews, which matched activity peaks in wild tree shrews. These results provided basal physiological indexes for domesticated tree shrews and laid an important foundation for diabetes and stress-related disease models established on tree shrews.

Smad3 knock-out mice as a useful model to study intestinal fibrogenesis

AIM： To evaluate the possible differences in morphology and immunohistochemical expression of CD3, transforming growth factor 131（TGF-131）, Smad7, α-smooth muscle actin （α-Sma）, and collagen types Ⅰ-Ⅶ of small and large intestine in Smad3 null and wild-type mice. METHODS： Ten null and ten wild-type adult mice were sacrificed at 4 mo of age and the organs （esophagus, small and large bowel, ureters） were collected for histology（hematoxylin and eosin, Masson thrichrome, silver staining）, morphometry and immunohistochemistry analysis. TGF-β1 levels of intestinal tissue homogenates were assessed by ELISA. RESULTS： No macroscopic intestinal lesions were detected both in null and wild-type mice. Histological and morphometric evaluation revealed a significant reduction in muscle layer thickness of small and large intestine in null mice as compared to wild-type mice. Immunohistochemistry evaluation showed a significant increase of CD3＋T cell, TGF-β1 and Smad7 staining in the small and large intestine mucosa of Smad3 null mice as compared to wild-type mice. α-Sma and collagen Ⅰ-Ⅶ staining of small and large intestine did not differ between the two groups of mice. TGF-β1 levels of colonic tissue homogenates were significantly higher in null mice than in wildtype mice. In preliminary experiments a significant reduction of TNBS-induced intestinal fibrosis was observed in null mice as compared to wild-type mice.

Generation and characterization of a transgenic mouse model for pancreatic cancer

AIM： To generate a SV40Tag transgenic tumor animal model and to study the mechanism underlying tumorigenesis. METHODS： A mammary gland expression vector containing SV40Tag DNA was generated. Transgene fragments were microinjeted into fertilized eggs of FVB mice. The genetically manipulated embryos were transferred into the oviducts of pseudo-pregnant female mice. PCR and Northern blot analysis were used for genotype analysis of F1 and F2 mice. Transgene expression was detected by RT-PCR and immunohistochemistry. RESULTS： SV40Tag gene was detected in two lines of transgenic mice. One of them delivered the transgene to F1 and a tumor was found in the pancreas of these mice. RT-PCR and immunohistochemistry showed that SV40Tag gene was expressed in the tumor. Pathological characterization of the transgenic mice demonstrated that the tumor belonged to pancreatic cystic neoplasm. CONCLUSION： SV40Tag transgenic mouse model can be successfully established. The transgenic mice develop a pancreatic tumor, which can be used for investigation of the molecular mechanism of tumorigenesis in vivo.

Systematic review of robust experimental models of rheumatoid arthritis for basic research

Rheumatoid arthritis(RA)is a common autoimmune disease characterized by progressive joint inflammation and destruction,deformity,loss of mobility,and permanent disability.Although the cellular and molecular mechanisms involved in RA are understood in detail,no drugs or therapies can completely cure RA.Many long-term efforts have been directed towards a better understanding of RA pathogenesis and the development of new classes of therapeutics.Thus,the ongoing elucidation of pathogenic events underlying RA mostly relies on studies of animal models.Herein,we comprehensively review and discuss the characteristics,challenges,and unresolved of issues of various experimental models of RA to provide a basis and reference for the rational selection of experimental RA models for basic investigations into traditional Chinese medicine(TCM).

A simple and sensitive method to assess ischemia occurrence in the setting of focal cerebral ischemia in rat: A comparative study

Objective: Neurological evaluation is commonly applied to identify ischemia in focal cerebral ischemia model though it might not be sensitive. In present study, we hired sleeping time to assess ischemia occurrence. Methods: Permanent middle cerebral artery occlusion was induced in Sprague-Dawley rats under pentobarbital and ketamine anesthesia respectively. Sleeping time was recorded. Neurological evaluation was conducted by modified Bederson’s scoring system at 4 h and histopathological evaluation was performed at 3 d after middle cerebral artery occlusion. Results: Slices of brain stained by TTC, H&E and hoechst 33258 revealed extensive lesion in the two ischemic groups. The sensitivity to identify ischemia by neurological evaluation was 62.5%, but it was 81.3% and 80% respectively when evaluating by sleeping time (pentobarbital: ≥90.7 min, ketamine: ≥36.1 min). The sensitivity to identify ischemia by sleeping time was significantly higher than that by neurological evaluation (P<0.05). Conclusion: Our results suggested that to identify ischemia by sleeping time is a simple and sensitive method in the setting of focal cerebral ischemia in rat.

A Macaca mulatta model of fulminant hepatic failure

AIM:To establish an appropriate primate model of fulminant hepatic failure (FHF).METHODS:We have,for the first time,established a large animal model of FHF in Macaca mulatta by intraperitoneal infusion of amatoxin and endotoxin.Clinical features,biochemical indexes,histopathology and iconography were examined to dynamically investigate the progress and outcome of the animal model.RESULTS:Our results showed that the enzymes and serum bilirubin were markedly increased and the enzyme-bilirubin segregation emerged 36 h after toxin administration.Coagulation activity was significantly decreased.Gradually deteriorated parenchymal abnormality was detected by magnetic resonance imaging (MRI) and ultrasonography at 48 h.The liver biopsy showed marked hepatocyte steatosis and massive parenchymal necrosis at 36 h and 49 h,respectively.The autopsy showed typical yellow atrophy of the liver.Hepatic encephalopathy of the models was also confirmed by hepatic coma,MRI and pathological changes of cerebral edema.The lethal effects of the extrahepatic organ dysfunction were ruled out by their biochemical indices,imaging and histopathology.CONCLUSION:We have established an appropriate large primate model of FHF,which is closely similar to clinic cases,and can be used for investigation of the mechanism of FHF and for evaluation of potential medical therapies.

Study of a Novel Small Caliber Vascular Graft in a Canine Model with Optical and Scanning Electron Microscopy

A novel biological small-diameter vascular graft was evaluated in a canine model. 3 cm long segments with 4 mm I.D. were implanted end-to-end in the carotid position of 12 dogs for 6 months. Color Doppler sonography was performed at the first week post-operation, and angiography was then administered to 9 grafts at 4th week, 12th week and 24th week respectively to monitor the graft pantency and blood flow characteristics. Vascular samples containing the grafts were collected at 1st week, 8th week, 12th week and 24th week after implantation. Morphological changes of the grafts were observed by optical and scanning electron microscopic (SEM) studies and compared with that of the original prosthesis and the normal host vessel. All grafts were patent throughout the experiment except one graft. Histopathology and SEM demonstrated both a nearly complete inner capsule of varied thickness lining the graft luminal surface and connective tissue adventitia formation at one-week post-operation. The neointima became confluent at 8 weeks and then compact but had no signs of hyperplasia up to 12 weeks; meanwhile on the neointimal surface newly grown endothelial-like cells were migrating from the stoma to the middle portion. The grafts also illustrated endothelialization in many “islands” in the mid-segment luminal surface of the grafts. In addition, the closer distance the cells towards the stoma were, the more morphological similarity the cells with the normal endothelial were. Taken together, the biological vascular graft remained patent for 24 weeks as a carotid prosthesis, characterized by the early and complete neointima formation plus endothelialization starting before 12 weeks post grafting. Therefore, the graft seems suitable for reconstruction of vascular lesions in dogs. Further studies may be carried out to extend the graft application for the clinical use.

THE PATHOGENESIS OF EXPERIMENTAL MODEL OF MITOCHONDRIAL MYOPATHY INDUCED BY GERMANIUM DIOXIDE

Objective. The purpose of the study was to build up an animal model of mitochondrial myopathy in order to analyse the pathogenesis of the disease. Methods. The skeletal muscles from Wistar rats treated with germanium dioxide for 24 weeks were analysed by histopathologic and electron- microscopic studies. A quantitative analysis was carried out in mitochondrial DNAs of these samples. The biological function of the model was determined. Results. An animal model of mitochondrial myopathy was built up, in which oxygen free radicals were increased and mitochondrial DNA copies were decreased contrasted with controls. Conclusion. It suggested that environmental toxin may play a role in the pathogenesis of mitochondrial myopathy. The increase of oxygen free radicals is an important link causing the disease.