六味地黄汤现代药理学及化学的初步研究

Liuwei Dihuang decoction（LW），a classical traditional Chinese medicinal prescription with the effect of “nourishing Kidney-Yin”，consists of six herbs including Rehmannia glutinosa (Shudi),Cornus officinalis(Shauzhuyu)，Dioscorea opposita(Shanyao)，Alisma orientale(Zexie)，Porie cocos（Fuling）and Paeonia suffruticosa（Mudanpi）．It has long been used clinically in the treatments of many kinds of diseases with the sign of “Kidney-Yin deficiency”，such as cancer，diabetes，autoimmue diseases，menopausal syndrome in traditional Chinese medical treatments．The effectiveness of it has been well documented during the long-term clinical practices．In our study，the effects and the possible active mechanisms of LW on learning and memory ，neuroendocrine and immune functions were studies from the angle of neuroendocrine immunomodulation（NIM）network ．In chemical fractionation and purification of the immunoactive fractions or components contained in LW，the method of a close collaboration between pharmacological and chemical studies was employed．The chemical fractionation was begun from the decoction and guided by pharmacological activity evaluation．The results showed that oral administration of LW significantly improved learning and memory abilities in several kinds of model animals such as senescence accelerated mice (SAM)，chronic hanging stress-loaded mice and corticosterone-treated mice and the mechanism studies suggested that the effects were achieved through several ways．LW significantly decreased the plasma level of corticosterone in SAM-prone/8 (SAMP8) ，a substrain of SAM．SAMP8 showed a marked elevation of plasma level of corticosterone with advancing age in comparison with SAMR1 ，a substrain of senescence resistant of SAM．LW also significantly improved the secretion of testosterone by primary cultured testis cells from SAMP8．The immunopharmacological studies showed that oral administration of LW not only improved the immune functions in several immunodeficient model animals such as cyclophospharmide-treated mice and SAMP8，but also modulated or corrected the imbalance of immune functions manifested in autoimmune model animals includig adjuvant arthritis rats and campylobacter jejuni primed mice．Under the guidance of activity evaluation，several immunoactive fractions and pure compounds were obtained from the decoction．The effects of some fractions and compounds were studied and the results showed that they possessed potential immunopharmacological effects．The preliminary study suggested that LW possessed wide pharmacological effects and the basic effect of LW，we postulate，is modulation the balance of NIM network disturbed in many pathological conditions．It is also suggested that our method employed in the study of LW is an effective and practicable way in modern studies of tradltional Chinese medicinal prescriptions．

Objective To evaluate the use of the saphenous artery as an alternative access for endovascular procedures in the porcine model. Methods Fourteen adult pigs (25-35 kg) were used in this study, 3 pigs from an acute study and 11 from chronic studies. A 2-3 cm incision was made and a saphenous artery cutdown was performed in 24 sides. Micropuncture sets (Boston Scientific) or 18 G puncture needles were used to access the artery. Different sizes of introducers (4-7 F) were used to establish endovascular access. Angiographic catheters were then used to confirm if the access was usable. Four saphenous arteries were explanted in the pigs from a chronic study 4 to 28 days after surgical procedure. Results The saphenous artery was very easy to expose and 4-5 F introducer sheaths were able to be inserted to establish access for endovascular procedures in the pigs. The saphenous artery was unable to accomodate an introducer with a size larger than 6 F. Four saphenous arteries were injured when 5 and 6 F introducers were used, but angiographic procedures could still be performed. Morphologic evaluation of the explanted arteries demonstrated occlusion of the saphenous arteries without injury or disruption of the adjacent femoral arteries. Conclusion The saphenous artery can be used as an access site in pigs for angiographic and interventional procedures if the catheter size is less than 6 F. This vessel is easier to access and can preserve the femoral artery for repeat procedures in the future.

Objective To evaluate the utility of multi-slice computed tomography (MSCT) in assessing acute non-reperfused myocardial infarct size. Methods Seven domestic pigs (mean weight 17.3 ± 1.9 kg) underwent ligation of the distal left anterior descending artery to establish a model of acute myocardial infarction (MI). MSCT and triphenyltetrazolium chloride (TTC) staining were performed two hours later. The following data were acquired and analyzed:MI volume (%), CT values of the infarcted region, left ventricular cavity and normal cardiac tissue at various scanning time-points (1, 5, 10, 15, 20 min after contrast injection). Results Using MSCT, the overall MI volume showed a time-dependent decrease, with a reduction of 28.87%after 20 min. The greatest reduction occurred at the 5 min time-point. In TTC staining, MI volume was 9.87%± 2.44%. When MI size, as determined by MSCT, was compared with that by TTC staining in Bland-Altman plots, there was a better agreement at 5, 10, and 15 min time-points at 1 and 20 min. Conclusions The study indicates that double-phase scanning examination using MSCT is a useful tool to assess MI size, and the optimal late-phase scanning time-point set within 5-15 min of contrast injection.

新型室间隔缺损封堵器的动物实验研究

心房颤动动物模型制备

心房颤动(房颤)是临床常见的心律失常,也是增加心血管病致残率和致死率的重要因素,至今仍未能阐明其确切机制.对房颤模型的深入研究为我们提供了许多重要认识.房颤模型也由初的心房肌组织片计算机模型和Lan-gendoff灌注离体心脏房颤模型发展到现今的在体房颤动物模型.本文就目前房颤动物模型的制备作一综述.

骨肉瘤动物模型的构建及其应用现状

骨肉瘤是一种具有高度局部复发和远处转移倾向的原发性恶性骨肿瘤,多见于儿童及青少年[1].目前,尽管采用了加强化疗和适当的手术切除,仍有30%﹪～40%﹪的患者死于该疾病,其中肺转移是主要原因[2].建立接近人类骨肉瘤临床特征的肿瘤动物模型,对于研究骨肉瘤及寻找新的治疗方法具有重要的现实意义.

Objective To undertake animal experimentation and clinical study on the safety and efficacy of percutaneous transluminal angioplasty (PTA) and intraarterial papaverine (IAP) infusion for treatment of refractory symptomatic cerebral vasospasm (CVS). Methods In the experimental study, vasospasm was induced in rabbits by double injections of blood into the cisterna magna, IAP infusion was given on either the 4th day or the 7th day after occurrence of subarachnoid hemorrhage (SAH), and then neurological observation, angiography, light and electron microscopy were done. In the clinical study, since September 1996, 22 patients with refractory symptomatic CVS involving 50 vascular territories received dilation therapy by PTA and IAP within 24 hours of clinical neurological deterioration. Results In the experimental study, all the rabbits except two in the 'the 4th day' group showed angiographic dilation in all of the spastic basilar arteries, and neurological improvement; in the ' the 7th day' group angiographic dilation appeared in 4 (57. 1% ) out of 7 rabbits. After 24 hours, 1 rabbit in each group had recurrence of neurological deficits and angiographic constriction. In the clinical study after aneurysm clipping or endovascular coil embolization was done, within 72 hours of SAH all patients underwent endovascular treatment: PTA alone in 3 cases, IAP alone in 14 cases, PTA and IAP in the remaining 5 cases. All vessel segments were dilated satisfactorily after endovascular treatment. Clinical improvement was significant in 13 cases,moderate in 7, minimal or none in 2; 2 cases died on the 7th day after endovascular dilation treatment. Conclusion Endovascular dilating techniques, namely, PTA, IAP and a combination of PTA and IAP, are safe and effective for treatment of symptomatic CVS refractory to medical therapy.

A brachial plexus injury model was established in rabbits by stretching the C6 nerve root. Imme-diately after the stretching, a suspension of human amniotic epithelial cells was injected into the injured brachial plexus. The results of tensile mechanical testing of the brachial plexus showed that the tensile elastic limit strain, elastic limit stress, maximum stress, and maximum strain of the injured brachial plexuses were signiifcantly increased at 24 weeks after the injection. The treat-ment clearly improved the pathological morphology of the injured brachial plexus nerve, as seen by hematoxylin eosin staining, and the functions of the rabbit forepaw were restored. These data indicate that the injection of human amniotic epithelial cells contributed to the repair of brachial plexus injury, and that this technique may transform into current clinical treatment strategies.

Neural stem cells are characterized by the ability to differentiate and stably express exogenous ge-nes. Vascular endothelial growth factor plays a role in protecting local blood vessels and neurons of newborn rats with hypoxic-ischemic encephalopathy. Transplantation of vascular endothelial growth factor-transfected neural stem cells may be neuroprotective in rats with cerebral palsy. In this study, 7-day-old Sprague-Dawley rats were divided into ifve groups: (1) sham operation (control), (2) cerebral palsy model alone or with (3) phosphate-buffered saline, (4) vascular en-dothelial growth factor 165 + neural stem cells, or (5) neural stem cells alone. hTe cerebral palsy model was established by ligating the letf common carotid artery followed by exposure to hypox-ia. Phosphate-buffered saline, vascular endothelial growth factor + neural stem cells, and neural stem cells alone were administered into the sensorimotor cortex using the stereotaxic instrument and microsyringe. Atfer transplantation, the radial-arm water maze test and holding test were performed. Immunohistochemistry for vascular endothelial growth factor and histology using hematoxylin-eosin were performed on cerebral cortex. Results revealed that the number of vas-cular endothelial growth factor-positive cells in cerebral palsy rats transplanted with vascular endothelial growth factor-transfected neural stem cells was increased, the time for ifnding water and the ifnding repetitions were reduced, the holding time was prolonged, and the degree of cell degeneration or necrosis was reduced. hTese ifndings indicate that the transplantation of vascu-lar endothelial growth factor-transfected neural stem cells alleviates brain damage and cognitive deifcits, and is neuroprotective in neonatal rats with hypoxia ischemic-mediated cerebral palsy.

Endoplasmic reticulum stress is closely involved in the early stage of diabetic retinopathy. In the present study, a streptozotocin-induced diabetic animal model was given an intraperitoneal injection of tauroursodeoxycholic acid. Results from immunofluorescent co-localization experiments showed that both caspase-12 protein and c-Jun N-terminal kinase 1 phosphorylation levels significantly in-creased, which was associated with retinal ganglion celldeath in diabetic retinas. The C/ERB ho-mologous protein pathway directly contributed to glial reactivity, and was subsequently responsible for neuronal loss and vascular abnormalities in diabetic retinopathy. Our experimental findings in-dicate that endoplasmic reticulum stress plays an important role in diabetes-induced retinal neu-ronal loss and vascular abnormalities, and that inhibiting the activation of the endoplasmic reticulum stress pathway provides effective protection against diabetic retinopathy.

Because there is no curative treatment for spinal cord injury, establishing an ideal animal model is important to identify injury mechanisms and develop therapies for individuals suffering from spinal cord injuries. In this article, we systematically review and analyze various kinds of animal models of spinal cord injury and assess their advantages and disadvantages for further studies.

Objective: To establish an animal model of like-auditory neuropathy in neonatal rat. Methods The ani-mals were injected with phenylhydrazine hydrochloride or saline at 7-day of age. ABR and DPOAE were performed to assess the auditory function. The cochlea basilar membrane stretched preparation and cochlear frozen sections were prepared for immunohistochemical staining to examine the morphological change of hair cells and spiral ganglion cells (SGNs). Results At 7-day age the ABR waveI, III, V, latencies andI-III,I-V IWIs in the experimental group were significantly prolonged compared with those in the control group. The ABR thresholds were also elevated in the experimental group. We found there is no significant differ-ence in DPOAE in phenylhydrazine hydrochloride exposure group compare to control group. The cochlear hair cells showed no signs of loss in both group, but the total number of neurofilaments positive cells in SGNs were significantly reduced in the phenylhydrazine treated animals. Conclusion Our study suggests that phenylhydrazine hydrochloride can change the auditory function and induce peripheral nerve pathology by targeted mainly the SGNs in neonatal rat.

Objective To investigate the suitability of miniature pigs as an animal model for otological research. Methods Microdissection of the temporal bone was performed on 10 miniature pigs and recorded on photo-graphs. Results The morphology and measurement of the external, middle and inner ear and the lateral re-cess of the miniature pigs were obtained by microdissection. Conclusion Compared to traditional animal models, the miniature pig may be a better model for biomedical research because of its many similarities in physiological functions with humans. Similarities of the temporal bone structures, including the external, middle and inner ear and the lateral recess, between the miniature pig and human make the animal a poten-tially useful model for otological research.

缺氧预适应过程中的小鼠脑一氧化氮下调

Downregulation of nitric oxide in the brain of mice during their hypoxic preconditioning.J Appl Physiol 91:1193～1198,2001.An animal model of hypoxic preconditioning was produced in mice by repeated exposure to autohypoxic condition.The animals' tolerance times to hypoxia were 1.7,1.8,2.1 and 2.3 times longer in runs 2,3,4 and 5,respectively,than that in run 1,and their oxygen consumption and heart and respiration rates were progressively and significantly slowed down during the repetitive exposure to hypoxia.L-arginine concentration,nitric oxide (NO) synthase-positive cells,NO synthase activity,and NO content in the whole brain and the subregions telencephalon,diencephalons,and brain stem were significantly increased during the first exposure and were,instead of continuing to increase,significantly decreased in run 4 after the second and third exposure.Tolerance times under the hypoxic condition were significantly shortened and prolonged when preadministration of L-arginine and its analog,respectively,was made.These results indicate that NO in the brain is downregulated under condition of hypoxic preconditioning and negatively involved in increased tolerance to hypoxia.

从动物模型到个体化治疗——中山大学黄如训教授谈脑梗死的科研与临床策略

研究脑卒中的机制并制订合理的治疗方案是十分复杂和困难的,尤其是常见的脑梗死,直至目前尚无达成共识和普遍适用的治疗方案.多年来,国内外进行了大量有关脑梗死的基础和临床研究,为探索有效的治疗方法提供了依据.中山大学的黄如训教授在动物实验研究上取得了一系列的创新性成果,随后又提出了卒中应采取个体化治疗方案的观点,受到了国内外同行的关注.针对脑梗死的科研与临床策略问题,黄如训教授阐述了他的观点.

Ⅱ型糖尿病实验性动物模型和转基因动物模型的研究与应用

近年来,糖尿病发病率上升很快,其中Ⅱ型糖尿病(NIDDM)的发病率占糖尿病发病率的90%以上,合适的糖尿病模型无疑可对糖尿病研究提供重要的作用.本文对实验性Ⅱ型糖尿病模型研究的现状及相关资料综述如下.

自发性高血压大鼠行为学的初步观察

背景:注意缺陷多动障碍 (attention deficit hyperactivity disorder, ADHD)存在着强化机制的异常,而多动性是强化机制异常的一种表现.目的 :观察 ADHD动物模型自发高血压大鼠( SHR)的多动性变化,以探究其强化机制.设计:非随机对照的实验研究.地点、材料和干预:实验在挪威奥斯陆大学医学系生理实验室进行.实验选用 12只雄性 SHR大鼠和 12只雄性 Wistar Kyoto( WKY)大鼠.实验在标准 Campden连续反应测试箱中进行,大鼠禁水 22 h,采用长变异间期方案(变异指数 180 s),分析后 10 d稳定曲线的均值.主要观察指标:大鼠按杠次数、推门次数及在不同阶段的变化.结果 :① SHR组大鼠按杠次数 [(642.9± 173.1)次 ]显著多于 WKY组 [(219.9± 79.5)次〗 (t=7.52, P《 0.01).②实验过程的 3个相同阶段 SHR大鼠的按杠次数比较,差异无显著性意义 (F=0.10, P 》0.05).③ SHR组的推门次数明显多于 WKY组 (t=4.43, P《 0.01),而获水次数与 WKY组比较,差异无显著性意义 (t=1.86, P 》0.05).结论: SHR大鼠的多动性较 WKY大鼠强,强化机制异常,有助于了解 ADHD的发病机制、临床治疗及预防.

Usefulness of animal models for tuberculosis research-A review

Preparation animal model of intracristal and membranous ventricular septal defects\室间隔缺损封堵治疗的疗效和安全性

先天性胆道扩张症动物模型的制作——P-Ch吻合术的实践与结果

作者从事先天性胆道扩张症的病因学研究,采用日本学者大川氏的手术方法成功地制造了狗的动物模型,并进行了有关X线学、生化学、病理学的研究.本文还就本症病因学研究的各种方法进行了讨论.