中国恒河猴艾滋病病证复合模型创建及治疗艾滋病中药复方优化研究
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摘要
目的：
国内中医药防治艾滋病的基础研究相对于西医药防治艾滋病的同类研究起步晚,目前尚属探索阶段。利用适宜的动物艾滋病模型进行艾滋病研究具高度的可控性、可靠性、可重复性,同时可缩短研究进程,减少干扰因素,降低试验风险。在某种程度上来说艾滋病研究中,采用动物艾滋病模型研究比采用人体试验进行研究更易实现。国内在上世纪90年代后期已经开始利用动物模型进行艾滋病的研究,但具有中医证特征的艾滋病动物模型研究方面属于空白,这对中医药治疗艾滋病疗效的科学客观评价带来了明显障碍。恒河猴艾滋病模型为艾滋病研究中动物模型的金标准,并被世界卫生组织WHO所推荐。国外研究多采用印度恒河猴,但由于印度恒河猴目前数量减少、价格昂贵、出口受严格限制,已经不能满足日益增长的科研需求,这给开展艾滋病的深入研究带来了明显阻碍。越来越多的研究采用来源相对充足的中国恒河猴替代印度恒河猴进行艾滋病研究。本研究在国家“十一五”科技重大专项资助下,对中国恒河猴艾滋病模型以及具中医证特征的中国恒河猴艾滋病模型开展深入详细研究,并利用这些动物艾滋病模型进行中医药治疗艾滋病方面的研究。希望通过上述研究后实现以下的目标：(1)通过开展对中国恒河猴实验基础规范的标准化制定及基础背景的研究,进而深入研究中国恒河猴艾滋病模型/病证复合模型,深化中医对艾滋病的认知；(2)在中医理论指导下采用猴艾滋病病证复合模型观察中药干预的效果,探讨中医理论对艾滋病治疗的意义；(3)采用猴艾滋病模型对中药进行疗效优化,为进一步进行新药开发提供实验依据；(4)通过中西药代谢性相互作用研究,为进一步探讨中西医结合治疗艾滋病的合理性提供范例及实验依据。
方法：
1、文献研究：使用ENDNOTE软件收集国内外主流医学杂志文献,详细阅读后剔除重复或者资料不齐全文献,对文献进行整理分类和有效信息集中,最后选择有代表性文献,进行总结归纳,了解现代医学在艾滋病研究方面的最新状况。文献研究主要集中在三个方面,具体为艾滋病西医研究进展方面、艾滋病中医研究进展方面、动物疾病模型在中医药研究中的应用进展方面。在完成文献研究后提出本实验研究的整体设计思路。
2、中国恒河猴实验基础规范的标准化制定及基础背景研究：本部分研究将进行各项实验基础规范化的研究,保证实验研究结果的准确、可靠、可重复性；同时对中国恒河猴基础背景进行研究,深入了解了动物模型的研究背景。研究具体分为中国恒河猴艾滋病动物模型实验研究相关标准操作／作业流程(SOP)制定、健康中国恒河猴指标参考范围的研究两大部分。其中中国恒河猴艾滋病动物模型实验研究相关标准操作／作业流程制定部分为主要针对中国恒河猴动物试验、检测、操作、观察规范化进行标准制定；而健康中国恒河猴指标参考范围的研究部分采用健康中国恒河猴横断面调查研究,制定在本试验研究条件下多项检测指标正常参考范围。
3、中国恒河猴艾滋病动物模型研究：本部分研究主要针对中国恒河猴艾滋病动物模型进行深入细化研究,研究涉及猴艾滋病的疾病各阶段。研究第一步是对急性感染过程中体质量指标、T淋巴细胞亚群指标、病毒载量指标、血常规指标等多项指标的变化情况进行分析比较；研究第二步对长期感染过程中T淋巴细胞亚群指标、病毒载量指标、中医证候学指标以及临床症状变化情况等进行长期序贯性监测；研究第三步是对中国恒河猴艾滋病动物模型生存状况的调查；研究第四步是综合前三步试验结果对猴艾滋病模型进行中医基本病因病机的探索。通过这些研究试图找到疾病变化的规律特征,在此基础上为中医药治疗艾滋病提供理论指导。
4、中国恒河猴艾滋病病证复合模型研究：本部分研究主要针对具中医证特征的中国恒河猴艾滋病动物模型进行深入细化研究。研究的内容为创建具中医脾虚证特征的中国恒河猴艾滋病病证复合模型和具中医肾虚证特征的中国恒河猴艾滋病病证复合模型,通过试验对比分析病证复合模型的指标变化规律特征,同时比较中医疗法中的补肾疗法、补脾疗法、清热疗法在艾滋病治疗中的疗效。并对各种疗法的优点和特点进行汇总分析,对艾滋病治疗方案提出优化后的具体方药。
5、研究成果拓展应用：本部分研究利用前面研究的成果进行了拓展应用。利用总结出的基本病因病机选择药物测试艾滋病晚期状态下中医药干预的效果；通过中西药代谢性相互作用研究,测试实验进程中优化后的中医药组方对植物单体化学物/西药在各种模型及健康志愿者中的增效作用。
结果：
1、中医药防治艾滋病疗效显著,不易产生耐药性,毒副反应较少或较轻。恒河猴艾滋病模型是研究艾滋病的动物模型“金标准”,广泛应用于艾滋病研究的各个方面。中医药研究中广泛使用动物模型进行具中医证特征的动物模型构建。利用中国恒河猴创建具中医证特征的艾滋病动物模型,进行中医药治疗艾滋病疗效的科学客观评价是必要和可行的方案。研究整体设计思路第一步为中国恒河猴实验基础规范的标准化制定及基础背景的研究；第二步为中国恒河猴艾滋病动物模型研究；第三步为中国恒河猴艾滋病病证复合模型研究；第四步为利用不同的动物艾滋病模型进行抗艾滋病中药复方优化的研究。
2、在中国恒河猴实验基础规范的标准化制定及基础背景的研究中,首先制定出1、恒河猴动物试验SOP；2、基础研究试验检测SOP；3、基础研究观察SOP；4、动物模型构建SOP；5、试验药物制作及使用SOP；6、恒河猴疾病防治SOP；7、试验数据处理SOP。对健康中国恒河猴研究后确立了血常规13项、肝肾功能4项、中医证学相关12项、T淋巴细胞亚群7项等共36个监测指标的正常参考范围。
3、对中国恒河猴艾滋病模型基础背景进行了深入的研究。对艾滋病急性感染期病毒载量、T淋巴细胞亚群等指标变化分析表明：中国恒河猴艾滋病模型血浆病毒峰值在10～14天,随后病毒载量整体出现波动下降的趋势。而T淋巴细胞亚群中CD4+%呈现下降趋势,CD4+/CD8+比值感染后在病毒载量高峰时期出现最低值,随后出现上升趋势。CD8+%则在急性感染阶段出现上升趋势。对中国恒河猴艾滋病模型感染18个月长期观察显示：T淋巴细胞亚群CD4+%、CD4+/CD8+比值随着感染时间延长呈现波动下降趋势；病毒载量在感染后2-3个月进入病毒调定点,此后病毒载量一般在病毒调定点上下0.5个对数波动；而中医证学相关指标皮质醇(Cortisol, Cor)在感染后随着感染时间延长呈现波动下降趋势。对因艾滋病死亡的中国恒河猴艾滋病模型数据分析表明：CD4+%、CD4+/CD8+比值、CD4+计数与病情密切相关；T3指标下降程度与病情恶化程度密切相关,Cor水平则在死亡前1-2个月出现上升趋势。对中国恒河猴艾滋病模型生存情况分析表明采用SIVmac239病毒株创建的猴艾滋病模型中位生存时间为2年左右。对艾滋病动物模型中医基本病因病机探索结果表明：本病的病因为“艾邪”。模型在急性感染期病机表现为外邪侵扰,邪正相争；在病毒潜伏期表现为邪毒内伏,暗伤正气；艾滋病前期表现为邪气亢盛,正气亏损；典型艾滋病期表现为正气亏虚。
4、通过不同的复合模型构建方案,试验成功首创了两种共四类具中医证特征的艾滋病病证复合模型。其中两种病证复合模型为中国恒河猴脾虚艾滋病病证复合模型和中国恒河猴肾虚艾滋病病证复合模型。四类为脾虚Ⅰ型病证复合模型(简称脾虚Ⅰ型)、脾虚Ⅱ型病证复合模型(简称脾虚Ⅱ型)、肾虚工型病证复合模型(简称肾虚Ⅰ型)、肾虚Ⅱ型病证复合模型(简称肾虚Ⅱ型)。其中中国恒河猴脾虚艾滋病病证复合模型临床症状表现出摄食减少、体质量增长缓慢、腹泻等,血液学指标三碘甲腺原氨酸(T3)、CD4+%、CD4+/CD8+比值水平均显著下降,而中国恒河猴肾虚艾滋病病证复合模型临床症状表现出畏寒、精神萎靡、消瘦、腹泻等,血液学指标三碘甲腺原氨酸(T3)、皮质醇(Cor)、CD4+%、CD4+/CD8+比值水平均显著下降。对四类病证复合模型深入研究后发现脾虚工型模型在复合模型创建阶段中建模因素对艾滋病病情影响不显著,但建模后造模因素对治疗阶段病情产生显著影响,能加速艾滋病病情的进展。在此类艾滋病模型中补脾法疗效优于清热法疗效；脾虚Ⅱ型模型研究结果显示脾虚Ⅱ型与脾虚Ⅰ型病情进展相类似,但比脾虚Ⅰ型病情更为严重,但在脾虚Ⅱ型试验中补脾法疗效并不优于清热法；肾虚Ⅰ型模型研究结果显示复合模型创建阶段中建模因素对艾滋病病情影响不显著,但建模后造模因素对治疗及停药期影响显著,能加速病情的进展。肾虚Ⅰ型试验中补肾法疗效优于清热法疗效,特别是补肾法在停药后8个月疗效与清热法差异非常显著；肾虚Ⅱ型模型研究结果显示肾虚Ⅱ型比肾虚Ⅰ型病情严重、肾虚Ⅱ型在复合模型创建期间造模因素对艾滋病病情影响显著,肾虚Ⅱ型试验中补肾法疗效优于清热法、肾虚Ⅱ型非治疗情况下病情进展更加迅速。在总结艾滋病动物模型/中国恒河猴艾滋病病证复合模型中医干预的经验基础上,针对猴艾滋病的基本病机,对原补肾处方进行药物优化,优化出新的补肾治疗方药(优化后组方)。
5、在研究成果拓展应用中,在艾滋病中医病因病机探索研究而归纳出的结论指导下,采用大补肾(元)阳药物参附注射液对中国恒河猴艾滋病晚期模型进行干预,成功的延长了模型的生存时间,减轻了临床症状,延缓了病情的进展。另外对优化后组方联合植物单体化学物治疗猴艾滋病动物模型的结果显示：中药和植物单体化学物E联用能明显增强植物单体化学物E的疗效,同时无明显毒副作用,具有良好的新药开发前景。进一步进行的优化后组方对猴艾滋病动物模型/健康志愿者服用蛋白酶抑制剂茚地那韦血药浓度影响的试验提示：优化后组方和茚地那韦联用能明显增加茚地那韦血药浓度,具减缓健康志愿者茚地那韦清除率作用。
结论：
1、文献研究表明了中医药在艾滋病防治方面具有明显疗效,对中医药防治艾滋病方面进行深入研究可能会取得较大的进展或突破。明确了在中医药防治艾滋病研究中采用适宜的恒河猴艾滋病动物模型进行研究比采用人体试验更容易实现。而且动物试验研究具高度可控、可靠、可重复性,可缩短研究进程,减少干扰因素,降低试验风险。明确了中国恒河猴创建具中医证特征的艾滋病病证复合模型的理论可行性。
2、中国恒河猴实验基础规范的标准化制定为本项目各试验的规范化研究提供了保证。制定出健康中国恒河猴多项指标正常参考范围,为实验结果的准确判断提供了相关试验依据。
3、中国恒河猴艾滋病动物模型能较好反应艾滋病病情变化特征,可适用于艾滋病多种类型的研究中。中国恒河猴艾滋病模型艾滋病的病因为艾邪,疾病过程病机变化整体表现为艾邪侵扰,邪正相争,随后邪毒内伏,暗伤正气,之后邪气亢盛,正气亏损,最后元气亏虚,五脏六腑气血阴阳亏竭。
4、具中医证特征的艾滋病病证复合模型因模型不一样,在艾滋病病情进展方面和中药干预效果方面也不完全一致。试验整体显示出具中医证特征的艾滋病病证复合模型病情比单纯的艾滋病模型严重。治疗效果比较中表明采用中药治疗远远优于不治疗效果,其中补肾疗法优于清热疗法,而补脾疗法与清热疗法在不同艾滋病动物模型中疗效不同。
5、研究成果的拓展应用证实了中医药在艾滋病治疗中的疗效,也再次证实了中西医联合治疗艾滋病的益处。对从理论上说明中药联合植物单体化学物／西药治疗艾滋病的合理性提供了范例和研究思路。
Objective
The basic research on treating AIDS with TCM in China, nowadays in its early stage, is conducted later than that of modern medicine. Research on AIDS with proper animal models is a controllable, reliable and repeatable procedure which can shorten the research process, reduce the confounding factors, and decrease the test risk. Therefore, to some degree, it is much easier to use animal AIDS models to do AIDS research rather than human trial. Although Chinese scholars have done the researches from late1990s, research on animal models with TCM symptoms is still in blank state, which is an obvious barrier to the scientific evaluation of efficacy of TCM on the treatment of AIDS. Rhesus monkey model for AIDS becomes a gold standard for researches on AIDS, and is recommended by WHO. Studies abroad usually use India rhesus monkey. However, due to the fact that the number of the India rhesus monkey deceases and they are expensive and strictly restricted to export, they can no longer meet the increasing needs of the scientific research and inhibits the in-depth research on AIDS. Therefore, more and more scholars use sufficient Chinese rhesus monkey to do their research instead of India rhesus monkey. With the fund by "Eleven-Five" national technology support plans, the study investigated a detailed study on Chinese rhesus monkey model for AIDS, including characteristics of healthy monkeys, characteristics of Chinese rhesus monkey model for AIDS and composite model of Chinese rhesus monkey for AIDS symptoms, treatment of rhesus monkey with TCM, and application of the research results. The targets of this case study include:(1) to study the rhesus monkey AIDS model/Syndrome composite model and deepen the awareness of TCM for AIDS through the basic research of the Chinese rhesus monkey;(2) to explore the significance of the theory of Chinese medicine for AIDS treatment by observing effect of TCM intervention under the guidance of TCM theory through composite model of the monkey AIDS syndromes;(3) to optimize the efficacy of Chinese medicine through monkey AIDS model and provide the experimental basis for the further development of new drugs;(4) to study the metabolic interaction of Chinese and Western medicine and provide some examples and experimental evidence to the reasonability of treating AIDS with the combined TCM and Western medicine.
Methods
1. Theoretical researches and literature review
The software of ENDNOTE was used to collect related literature in mainstream medical journals at home and abroad. After close reading of all the literature, we deleted the duplicate or incomplete information, classified the effective literature, and chose the representative ones to summarize the information to understand the latest researches on AIDS and put forward our own ideas. The literature review mainly focused on the following three aspects: the advancement of research on AIDS in modern medicine, the advancement of research on AIDS in TCM, and the advancement of research on animal models. The overall research design was proposed after the literature review.
2. Standardization of experimental basis of Chinese rhesus monkey and research on its background
In this part, the standardization of basic researches was done to ensure that the results are accurate, reliable and repeatable. Meanwhile, the background of Chinese rhesus monkey was studies to strengthen the understanding the research background of animal model. They can be divided into two major parts:the establishment of standard procedures of empirical study on Chinese rhesus monkey model for AIDS (Standard Operating Procedure in the lab), and research on the referrence ranges of indicators about the healthy Chinese rhesus monkey. The former was mainly about the establishment of standard on the experiment, testing, operation and observation of the Chinese rhesus monkey, and the latter was about the normal referrence ranges of various indicators under the condition of this study.
3. Research on Chinese rhesus monkey model for AIDS
This part was the detailed study on the Chinese rhesus monkey model and it involved various stages of AIDS. There were mainly four steps in the study: Firstly, we compared the changes of T-lymphocyte subsets and viral load during the acute infectious period; Secondly, we monitored the changes of T-lymphocyte subsets, viral load, hematological variables in TCM-syndrome and clinical symptoms regularly and sequentially; Thirdly, we investigated the survival condition of the Chinese rhesus monkey model and finally we explored the TCM pathogenesis of the disease based on the results of the previous three steps. The purpose of these researches was to find out the characteristics of the changes of the disease and provide some theoretical guidance for treating AIDS with TCM.
4. Research on composite model of Chinese rhesus monkey for AIDS symptoms
This part was the detailed study on Chinese rhesus monkey model with TCM symptoms. The key point of the study was to build composite model of Chinese rhesus monkey for AIDS with symptom of spleen-deficiency and composite model of Chinese rhesus monkey for AIDS with symptom of kidney-deficiency and compare the characteristics of the changes of the indicators through experiment. What's more, we compared the therapeutic effects of the TCM therapy like kidney-tonifying therapy, spleen-tonifying therapy and heat-releasing therapy in the treatment of AIDS. The advantages and characteristics of each therapy were analyzed and summarized to put forward an optimized TCM prescription for the treatment of AIDS.
5. Application of the research results
In this part, we applied the results of previous researches. Based on the summarized basic knowledge on pathogenesis, we selected drugs and tested the effect of TCM intervention in advanced AIDS period. We also tested the synergy of the refined and optimized TCM prescription on single plant chemicals/western medicine in various models and healthy volunteers through metabolic interaction studies on Chinese and Western medicine.
Results
1. TCM medicine is effective to prevent and treat AIDS, which is unlikely to develop the drug resistance and has a small or mild side effect. Rhesus monkey model for AIDS is the golden standard of the animal models for AIDS, and it is widely used in AIDS research. In the TCM research, animal models are used to establish the TCM-symptoms model, so it is necessary and feasible to establish the Chinese rhesus monkey model to evaluate the efficacy of TCM on the treatment of AIDS. The overall research design consisted of four steps: step1, standardization of experimental basis of Chinese rhesus monkey and research on its background; step2, research on Chinese rhesus monkey model for AIDS; step3, research on composite model of Chinese rhesus monkey for AIDS symptoms; and step4, research on the optimization of anti-AIDS TCM prescription through different animal AIDS model.
2. During the standardization of experimental basis of Chinese rhesus monkey and research on its background, the Standard Operating Procedure (SOP) on Chinese rhesus monkey model for AIDS was established. It includes SOP of the rhesus monkey experiment, SOP of the basic experimental testing, SOP of basic experimental observation, SOP of building the animal model, SOP of experimental drug production and use, SOP of disease prevention in rhesus monkey and SOP of data analysis. Furthermore, the normal referrence ranges of36indicators (including13items about routine blood test,4items about liver and kidney function,12items about TCM symptoms and7items about T-lymphocyte subset) were defined after the research on healthy Chinese rhesus monkey.
3. A long-term in-depth study on Chinese rhesus monkey model on AIDS was conducted. Analyzing the changes of indicators like T-lymphocyte subset and viral load in acute period, we found plasma viral load reached its peak in10to14days and then exhibited fluctuating decrease trend as a whole, while the percent of CD4+in T-lymphocyte subset decreased and the ratio of CD4+/and CD8+was at its lowest when viral load reached its peak and then the ratio increased. As for the percent of CD8+,it increased during the whole acute period. The18-month-observation to the monkey model indicated that the percent of CD4+and the ratio of CD4+and CD8+exhibited fluctuating decrease trend as the infection progressed, whereas the viral load fluctuated around the set point by0.5logarithmic fluctuations. The variable related to TCM-syndrome like Cor exhibited fluctuating decrease trend as the infection progressed. The analysis of the data about the monkey model between AIDS period and death showed that the CD4+%, CD8+%, the value of CD4+/CD8+, and CD4+counts were closely related to the disease condition, but the decrease of T3was closely related to the deterioration of the disease, while the Cor level increased lto2months before death. The analysis of the survival condition of the animal model indicated that the median survival time of the monkey model infected with SIVmac239was about2years. The exploration of TCM pathogenesis through AIDS model showed that the cause of AIDS was Ai-pathogen, and the pathogenesis manifested as the invasion of Ai-pathogen, the struggle between the Ai-pathogen and healthy qi in acute infection period, then the incubation of Ai-pathogen and injury to the healthy qi in incubation period, then excess of Ai-pathogen and insufficiency of healthy qi in pre-AIDS period, and finally the deficiency of vitality in typical AIDS period.
4. The study created two composite model of Chinese rhesus monkey for AIDS with symptoms of spleen-deficiency and two composite model of Chinese rhesus monkey for AIDS with symptoms of kidney-deficiency successfully for the first time. That is spleen-deficiency type I, spleen-deficiency type II, kidney-deficiency type I, and kidney-deficiency type II. The clinical symptoms of spleen-deficiency composite model include decreased food intake, slow growth of the body mass, diarrhea and significant decline of the ratio of hematological indicators like triiodothyronine(T3), CD4+%and CD4+/CD8+,whereas in the kidney-deficiency composite model, the clinical symptoms showed as chills, listlessness, weight loss, diarrhea and significant decline of the ratio of hematological indicators like triiodothyronine (T3), Cor, CD4+%and CD4+/CD8+. Detailed research found that in spleen-deficiency type I, the effect of modeling factors on the disease of AIDS was not significant in the composite model creation stage, but significant after composite model creation stage, which will accelerate the progression of AIDS disease. It also turned out that the therapeutic effect of spleen-tonifying therapy was better than that of heat-releasing therapy for this model. As for spleen-deficiency type II, it showed similarity with the spleen-deficiency type I, but the deficiency of the spleen was more serious and the therapeutic effect of spleen-tonifying therapy was not better than that of heat-releasing therapy. The researches on kidney-deficiency type I indicated that the effect of modeling factors on the disease of AIDS was not significant in model creation stage, but significant on treatment and withdrawal period after the model creation period, which will accelerate the progression of AIDS disease.. The therapeutic effect of kidney-tonifying therapy was better than that of heat-releasing therapy, especially the effect of kidney-tonifying therapy8month after withdrawal. With regard to kidney-deficiency type II, it was much serious than kidney-deficiency type I, and the modeling factors had significant effect on AIDS disease during the model creation stage. Meanwhile, the therapeutic effect of kidney-tonifying therapy was better than that of heat-releasing therapy and the disease with this model progressed quickly without treatment. Based on the summary of experience of TCM intervention through animal AIDS model/composite model of Chinese rhesus monkey for AIDS symptoms, we optimized the original kidney-tonifying prescription and put forward the new one (optimized prescription) according to the pathogenesis of AIDS in monkey models.
5. The research results can be applied in several aspects. Based on the researches results of the pathogenesis of AIDS, the use of Shenfu injection for treating Chinese rhesus monkey model in the late stage successfully prolonged the survival time of the diseased monkey, alleviated the clinical symptoms and slowed down the progression of the disease. In addition, the results of treating monkey AIDS model with the optimized prescription of combination of single plant chemicals showed that the combination of TCM and single plant chemical E can significantly enhanced the efficacy of single plant chemical E without any obvious side effect. The experimental study on the effect of protease inhibitors indinavir on the plasma concentration in monkey AIDS animal model/healthy volunteers indicated that the combination of optimized prescription and indinavir significantly increased the plasma concentration of indinavir and played a role in slowing down the clearing rate of indinavir in the healthy volunteers.
Conclusions
1. The literature review shows that TCM has a significant effect on the prevention and treatment of AIDS and indicates that the in-depth research of TCM on AIDS prevention and treatment may make big progress or have a breakthrough. It also clarifies the fact that it is easier to use proper animal AIDS models to do AIDS research rather than human trial and the researches with animal models are controllable, reliable and repeatable procedures which can shorten the research process, reduce the confounding factors, and decrease the test risk, which proves the feasibility of the establishment of composite model for ADIS with TCM symptoms by Chinese rhesus monkey.
2. The establishment of standard procedures of empirical study on Chinese rhesus monkey model for AIDS (Standard Operating Procedure in the lab) basically guarantees accuracy and reliability of the research results and lays foundations to the standardization of operation, testing and observation of the experiment on Chinese rhesus monkey. The definition of normal fluctuating ranges of indicators of Chinese rhesus monkey under healthy condition provides related criteria to the correct judgment of the results.
3. Chinese rhesus monkey model for AIDS can reflect the changes of disease condition of AIDS, which is applicable in the various researches on AIDS. According to the Chinese rhesus monkey model, the cause of AIDS was Ai-pathogen, and the pathogenesis manifested as the invasion of Ai-pathogen, the struggle between the Ai-pathogen and healthy qi, then the incubation of Ai-pathogen and injury to the healthy qi, then excess of Ai-pathogen and insufficiency of healthy qi, and finally the deficiency of vitality and exhaustion of five zang and six fu (the internal organs), qi blood and ying and yang.
4. The composite AIDS model with TCM symptoms varies, and they have different effect on the progression of AIDS and TCM intervention. The study indicates that the disease condition of the composite AIDS model with TCM symptoms is more serious than the simple AIDS model. The comparison of the therapeutic effect indicates that the treatment with TCM is much better than non-treatment, among which kidney-tonifying therapy is better than heat-releasing therapy and the therapeutic effect of spleen-tonifying therapy and heat-releasing therapy is different in different models.
5. The application of the results proves the therapeutic effect of TCM on the treatment of AIDS, and also the benefits of treating AIDS with combined drug use of TCM and modern medicine, which theoretically provides some examples and research ideas for the reasonability of treating AIDS with the combined TCM of single plant chemicals and Western medicine.
国内中医药防治艾滋病的基础研究相对于西医药防治艾滋病的同类研究起步晚,目前尚属探索阶段。利用适宜的动物艾滋病模型进行艾滋病研究具高度的可控性、可靠性、可重复性,同时可缩短研究进程,减少干扰因素,降低试验风险。在某种程度上来说艾滋病研究中,采用动物艾滋病模型研究比采用人体试验进行研究更易实现。国内在上世纪90年代后期已经开始利用动物模型进行艾滋病的研究,但具有中医证特征的艾滋病动物模型研究方面属于空白,这对中医药治疗艾滋病疗效的科学客观评价带来了明显障碍。恒河猴艾滋病模型为艾滋病研究中动物模型的金标准,并被世界卫生组织WHO所推荐。国外研究多采用印度恒河猴,但由于印度恒河猴目前数量减少、价格昂贵、出口受严格限制,已经不能满足日益增长的科研需求,这给开展艾滋病的深入研究带来了明显阻碍。越来越多的研究采用来源相对充足的中国恒河猴替代印度恒河猴进行艾滋病研究。本研究在国家“十一五”科技重大专项资助下,对中国恒河猴艾滋病模型以及具中医证特征的中国恒河猴艾滋病模型开展深入详细研究,并利用这些动物艾滋病模型进行中医药治疗艾滋病方面的研究。希望通过上述研究后实现以下的目标：(1)通过开展对中国恒河猴实验基础规范的标准化制定及基础背景的研究,进而深入研究中国恒河猴艾滋病模型/病证复合模型,深化中医对艾滋病的认知；(2)在中医理论指导下采用猴艾滋病病证复合模型观察中药干预的效果,探讨中医理论对艾滋病治疗的意义；(3)采用猴艾滋病模型对中药进行疗效优化,为进一步进行新药开发提供实验依据；(4)通过中西药代谢性相互作用研究,为进一步探讨中西医结合治疗艾滋病的合理性提供范例及实验依据。
方法：
1、文献研究：使用ENDNOTE软件收集国内外主流医学杂志文献,详细阅读后剔除重复或者资料不齐全文献,对文献进行整理分类和有效信息集中,最后选择有代表性文献,进行总结归纳,了解现代医学在艾滋病研究方面的最新状况。文献研究主要集中在三个方面,具体为艾滋病西医研究进展方面、艾滋病中医研究进展方面、动物疾病模型在中医药研究中的应用进展方面。在完成文献研究后提出本实验研究的整体设计思路。
2、中国恒河猴实验基础规范的标准化制定及基础背景研究：本部分研究将进行各项实验基础规范化的研究,保证实验研究结果的准确、可靠、可重复性；同时对中国恒河猴基础背景进行研究,深入了解了动物模型的研究背景。研究具体分为中国恒河猴艾滋病动物模型实验研究相关标准操作／作业流程(SOP)制定、健康中国恒河猴指标参考范围的研究两大部分。其中中国恒河猴艾滋病动物模型实验研究相关标准操作／作业流程制定部分为主要针对中国恒河猴动物试验、检测、操作、观察规范化进行标准制定；而健康中国恒河猴指标参考范围的研究部分采用健康中国恒河猴横断面调查研究,制定在本试验研究条件下多项检测指标正常参考范围。
3、中国恒河猴艾滋病动物模型研究：本部分研究主要针对中国恒河猴艾滋病动物模型进行深入细化研究,研究涉及猴艾滋病的疾病各阶段。研究第一步是对急性感染过程中体质量指标、T淋巴细胞亚群指标、病毒载量指标、血常规指标等多项指标的变化情况进行分析比较；研究第二步对长期感染过程中T淋巴细胞亚群指标、病毒载量指标、中医证候学指标以及临床症状变化情况等进行长期序贯性监测；研究第三步是对中国恒河猴艾滋病动物模型生存状况的调查；研究第四步是综合前三步试验结果对猴艾滋病模型进行中医基本病因病机的探索。通过这些研究试图找到疾病变化的规律特征,在此基础上为中医药治疗艾滋病提供理论指导。
4、中国恒河猴艾滋病病证复合模型研究：本部分研究主要针对具中医证特征的中国恒河猴艾滋病动物模型进行深入细化研究。研究的内容为创建具中医脾虚证特征的中国恒河猴艾滋病病证复合模型和具中医肾虚证特征的中国恒河猴艾滋病病证复合模型,通过试验对比分析病证复合模型的指标变化规律特征,同时比较中医疗法中的补肾疗法、补脾疗法、清热疗法在艾滋病治疗中的疗效。并对各种疗法的优点和特点进行汇总分析,对艾滋病治疗方案提出优化后的具体方药。
5、研究成果拓展应用：本部分研究利用前面研究的成果进行了拓展应用。利用总结出的基本病因病机选择药物测试艾滋病晚期状态下中医药干预的效果；通过中西药代谢性相互作用研究,测试实验进程中优化后的中医药组方对植物单体化学物/西药在各种模型及健康志愿者中的增效作用。
结果：
1、中医药防治艾滋病疗效显著,不易产生耐药性,毒副反应较少或较轻。恒河猴艾滋病模型是研究艾滋病的动物模型“金标准”,广泛应用于艾滋病研究的各个方面。中医药研究中广泛使用动物模型进行具中医证特征的动物模型构建。利用中国恒河猴创建具中医证特征的艾滋病动物模型,进行中医药治疗艾滋病疗效的科学客观评价是必要和可行的方案。研究整体设计思路第一步为中国恒河猴实验基础规范的标准化制定及基础背景的研究；第二步为中国恒河猴艾滋病动物模型研究；第三步为中国恒河猴艾滋病病证复合模型研究；第四步为利用不同的动物艾滋病模型进行抗艾滋病中药复方优化的研究。
2、在中国恒河猴实验基础规范的标准化制定及基础背景的研究中,首先制定出1、恒河猴动物试验SOP；2、基础研究试验检测SOP；3、基础研究观察SOP；4、动物模型构建SOP；5、试验药物制作及使用SOP；6、恒河猴疾病防治SOP；7、试验数据处理SOP。对健康中国恒河猴研究后确立了血常规13项、肝肾功能4项、中医证学相关12项、T淋巴细胞亚群7项等共36个监测指标的正常参考范围。
3、对中国恒河猴艾滋病模型基础背景进行了深入的研究。对艾滋病急性感染期病毒载量、T淋巴细胞亚群等指标变化分析表明：中国恒河猴艾滋病模型血浆病毒峰值在10～14天,随后病毒载量整体出现波动下降的趋势。而T淋巴细胞亚群中CD4+%呈现下降趋势,CD4+/CD8+比值感染后在病毒载量高峰时期出现最低值,随后出现上升趋势。CD8+%则在急性感染阶段出现上升趋势。对中国恒河猴艾滋病模型感染18个月长期观察显示：T淋巴细胞亚群CD4+%、CD4+/CD8+比值随着感染时间延长呈现波动下降趋势；病毒载量在感染后2-3个月进入病毒调定点,此后病毒载量一般在病毒调定点上下0.5个对数波动；而中医证学相关指标皮质醇(Cortisol, Cor)在感染后随着感染时间延长呈现波动下降趋势。对因艾滋病死亡的中国恒河猴艾滋病模型数据分析表明：CD4+%、CD4+/CD8+比值、CD4+计数与病情密切相关；T3指标下降程度与病情恶化程度密切相关,Cor水平则在死亡前1-2个月出现上升趋势。对中国恒河猴艾滋病模型生存情况分析表明采用SIVmac239病毒株创建的猴艾滋病模型中位生存时间为2年左右。对艾滋病动物模型中医基本病因病机探索结果表明：本病的病因为“艾邪”。模型在急性感染期病机表现为外邪侵扰,邪正相争；在病毒潜伏期表现为邪毒内伏,暗伤正气；艾滋病前期表现为邪气亢盛,正气亏损；典型艾滋病期表现为正气亏虚。
4、通过不同的复合模型构建方案,试验成功首创了两种共四类具中医证特征的艾滋病病证复合模型。其中两种病证复合模型为中国恒河猴脾虚艾滋病病证复合模型和中国恒河猴肾虚艾滋病病证复合模型。四类为脾虚Ⅰ型病证复合模型(简称脾虚Ⅰ型)、脾虚Ⅱ型病证复合模型(简称脾虚Ⅱ型)、肾虚工型病证复合模型(简称肾虚Ⅰ型)、肾虚Ⅱ型病证复合模型(简称肾虚Ⅱ型)。其中中国恒河猴脾虚艾滋病病证复合模型临床症状表现出摄食减少、体质量增长缓慢、腹泻等,血液学指标三碘甲腺原氨酸(T3)、CD4+%、CD4+/CD8+比值水平均显著下降,而中国恒河猴肾虚艾滋病病证复合模型临床症状表现出畏寒、精神萎靡、消瘦、腹泻等,血液学指标三碘甲腺原氨酸(T3)、皮质醇(Cor)、CD4+%、CD4+/CD8+比值水平均显著下降。对四类病证复合模型深入研究后发现脾虚工型模型在复合模型创建阶段中建模因素对艾滋病病情影响不显著,但建模后造模因素对治疗阶段病情产生显著影响,能加速艾滋病病情的进展。在此类艾滋病模型中补脾法疗效优于清热法疗效；脾虚Ⅱ型模型研究结果显示脾虚Ⅱ型与脾虚Ⅰ型病情进展相类似,但比脾虚Ⅰ型病情更为严重,但在脾虚Ⅱ型试验中补脾法疗效并不优于清热法；肾虚Ⅰ型模型研究结果显示复合模型创建阶段中建模因素对艾滋病病情影响不显著,但建模后造模因素对治疗及停药期影响显著,能加速病情的进展。肾虚Ⅰ型试验中补肾法疗效优于清热法疗效,特别是补肾法在停药后8个月疗效与清热法差异非常显著；肾虚Ⅱ型模型研究结果显示肾虚Ⅱ型比肾虚Ⅰ型病情严重、肾虚Ⅱ型在复合模型创建期间造模因素对艾滋病病情影响显著,肾虚Ⅱ型试验中补肾法疗效优于清热法、肾虚Ⅱ型非治疗情况下病情进展更加迅速。在总结艾滋病动物模型/中国恒河猴艾滋病病证复合模型中医干预的经验基础上,针对猴艾滋病的基本病机,对原补肾处方进行药物优化,优化出新的补肾治疗方药(优化后组方)。
5、在研究成果拓展应用中,在艾滋病中医病因病机探索研究而归纳出的结论指导下,采用大补肾(元)阳药物参附注射液对中国恒河猴艾滋病晚期模型进行干预,成功的延长了模型的生存时间,减轻了临床症状,延缓了病情的进展。另外对优化后组方联合植物单体化学物治疗猴艾滋病动物模型的结果显示：中药和植物单体化学物E联用能明显增强植物单体化学物E的疗效,同时无明显毒副作用,具有良好的新药开发前景。进一步进行的优化后组方对猴艾滋病动物模型/健康志愿者服用蛋白酶抑制剂茚地那韦血药浓度影响的试验提示：优化后组方和茚地那韦联用能明显增加茚地那韦血药浓度,具减缓健康志愿者茚地那韦清除率作用。
结论：
1、文献研究表明了中医药在艾滋病防治方面具有明显疗效,对中医药防治艾滋病方面进行深入研究可能会取得较大的进展或突破。明确了在中医药防治艾滋病研究中采用适宜的恒河猴艾滋病动物模型进行研究比采用人体试验更容易实现。而且动物试验研究具高度可控、可靠、可重复性,可缩短研究进程,减少干扰因素,降低试验风险。明确了中国恒河猴创建具中医证特征的艾滋病病证复合模型的理论可行性。
2、中国恒河猴实验基础规范的标准化制定为本项目各试验的规范化研究提供了保证。制定出健康中国恒河猴多项指标正常参考范围,为实验结果的准确判断提供了相关试验依据。
3、中国恒河猴艾滋病动物模型能较好反应艾滋病病情变化特征,可适用于艾滋病多种类型的研究中。中国恒河猴艾滋病模型艾滋病的病因为艾邪,疾病过程病机变化整体表现为艾邪侵扰,邪正相争,随后邪毒内伏,暗伤正气,之后邪气亢盛,正气亏损,最后元气亏虚,五脏六腑气血阴阳亏竭。
4、具中医证特征的艾滋病病证复合模型因模型不一样,在艾滋病病情进展方面和中药干预效果方面也不完全一致。试验整体显示出具中医证特征的艾滋病病证复合模型病情比单纯的艾滋病模型严重。治疗效果比较中表明采用中药治疗远远优于不治疗效果,其中补肾疗法优于清热疗法,而补脾疗法与清热疗法在不同艾滋病动物模型中疗效不同。
5、研究成果的拓展应用证实了中医药在艾滋病治疗中的疗效,也再次证实了中西医联合治疗艾滋病的益处。对从理论上说明中药联合植物单体化学物／西药治疗艾滋病的合理性提供了范例和研究思路。
Objective
The basic research on treating AIDS with TCM in China, nowadays in its early stage, is conducted later than that of modern medicine. Research on AIDS with proper animal models is a controllable, reliable and repeatable procedure which can shorten the research process, reduce the confounding factors, and decrease the test risk. Therefore, to some degree, it is much easier to use animal AIDS models to do AIDS research rather than human trial. Although Chinese scholars have done the researches from late1990s, research on animal models with TCM symptoms is still in blank state, which is an obvious barrier to the scientific evaluation of efficacy of TCM on the treatment of AIDS. Rhesus monkey model for AIDS becomes a gold standard for researches on AIDS, and is recommended by WHO. Studies abroad usually use India rhesus monkey. However, due to the fact that the number of the India rhesus monkey deceases and they are expensive and strictly restricted to export, they can no longer meet the increasing needs of the scientific research and inhibits the in-depth research on AIDS. Therefore, more and more scholars use sufficient Chinese rhesus monkey to do their research instead of India rhesus monkey. With the fund by "Eleven-Five" national technology support plans, the study investigated a detailed study on Chinese rhesus monkey model for AIDS, including characteristics of healthy monkeys, characteristics of Chinese rhesus monkey model for AIDS and composite model of Chinese rhesus monkey for AIDS symptoms, treatment of rhesus monkey with TCM, and application of the research results. The targets of this case study include:(1) to study the rhesus monkey AIDS model/Syndrome composite model and deepen the awareness of TCM for AIDS through the basic research of the Chinese rhesus monkey;(2) to explore the significance of the theory of Chinese medicine for AIDS treatment by observing effect of TCM intervention under the guidance of TCM theory through composite model of the monkey AIDS syndromes;(3) to optimize the efficacy of Chinese medicine through monkey AIDS model and provide the experimental basis for the further development of new drugs;(4) to study the metabolic interaction of Chinese and Western medicine and provide some examples and experimental evidence to the reasonability of treating AIDS with the combined TCM and Western medicine.
Methods
1. Theoretical researches and literature review
The software of ENDNOTE was used to collect related literature in mainstream medical journals at home and abroad. After close reading of all the literature, we deleted the duplicate or incomplete information, classified the effective literature, and chose the representative ones to summarize the information to understand the latest researches on AIDS and put forward our own ideas. The literature review mainly focused on the following three aspects: the advancement of research on AIDS in modern medicine, the advancement of research on AIDS in TCM, and the advancement of research on animal models. The overall research design was proposed after the literature review.
2. Standardization of experimental basis of Chinese rhesus monkey and research on its background
In this part, the standardization of basic researches was done to ensure that the results are accurate, reliable and repeatable. Meanwhile, the background of Chinese rhesus monkey was studies to strengthen the understanding the research background of animal model. They can be divided into two major parts:the establishment of standard procedures of empirical study on Chinese rhesus monkey model for AIDS (Standard Operating Procedure in the lab), and research on the referrence ranges of indicators about the healthy Chinese rhesus monkey. The former was mainly about the establishment of standard on the experiment, testing, operation and observation of the Chinese rhesus monkey, and the latter was about the normal referrence ranges of various indicators under the condition of this study.
3. Research on Chinese rhesus monkey model for AIDS
This part was the detailed study on the Chinese rhesus monkey model and it involved various stages of AIDS. There were mainly four steps in the study: Firstly, we compared the changes of T-lymphocyte subsets and viral load during the acute infectious period; Secondly, we monitored the changes of T-lymphocyte subsets, viral load, hematological variables in TCM-syndrome and clinical symptoms regularly and sequentially; Thirdly, we investigated the survival condition of the Chinese rhesus monkey model and finally we explored the TCM pathogenesis of the disease based on the results of the previous three steps. The purpose of these researches was to find out the characteristics of the changes of the disease and provide some theoretical guidance for treating AIDS with TCM.
4. Research on composite model of Chinese rhesus monkey for AIDS symptoms
This part was the detailed study on Chinese rhesus monkey model with TCM symptoms. The key point of the study was to build composite model of Chinese rhesus monkey for AIDS with symptom of spleen-deficiency and composite model of Chinese rhesus monkey for AIDS with symptom of kidney-deficiency and compare the characteristics of the changes of the indicators through experiment. What's more, we compared the therapeutic effects of the TCM therapy like kidney-tonifying therapy, spleen-tonifying therapy and heat-releasing therapy in the treatment of AIDS. The advantages and characteristics of each therapy were analyzed and summarized to put forward an optimized TCM prescription for the treatment of AIDS.
5. Application of the research results
In this part, we applied the results of previous researches. Based on the summarized basic knowledge on pathogenesis, we selected drugs and tested the effect of TCM intervention in advanced AIDS period. We also tested the synergy of the refined and optimized TCM prescription on single plant chemicals/western medicine in various models and healthy volunteers through metabolic interaction studies on Chinese and Western medicine.
Results
1. TCM medicine is effective to prevent and treat AIDS, which is unlikely to develop the drug resistance and has a small or mild side effect. Rhesus monkey model for AIDS is the golden standard of the animal models for AIDS, and it is widely used in AIDS research. In the TCM research, animal models are used to establish the TCM-symptoms model, so it is necessary and feasible to establish the Chinese rhesus monkey model to evaluate the efficacy of TCM on the treatment of AIDS. The overall research design consisted of four steps: step1, standardization of experimental basis of Chinese rhesus monkey and research on its background; step2, research on Chinese rhesus monkey model for AIDS; step3, research on composite model of Chinese rhesus monkey for AIDS symptoms; and step4, research on the optimization of anti-AIDS TCM prescription through different animal AIDS model.
2. During the standardization of experimental basis of Chinese rhesus monkey and research on its background, the Standard Operating Procedure (SOP) on Chinese rhesus monkey model for AIDS was established. It includes SOP of the rhesus monkey experiment, SOP of the basic experimental testing, SOP of basic experimental observation, SOP of building the animal model, SOP of experimental drug production and use, SOP of disease prevention in rhesus monkey and SOP of data analysis. Furthermore, the normal referrence ranges of36indicators (including13items about routine blood test,4items about liver and kidney function,12items about TCM symptoms and7items about T-lymphocyte subset) were defined after the research on healthy Chinese rhesus monkey.
3. A long-term in-depth study on Chinese rhesus monkey model on AIDS was conducted. Analyzing the changes of indicators like T-lymphocyte subset and viral load in acute period, we found plasma viral load reached its peak in10to14days and then exhibited fluctuating decrease trend as a whole, while the percent of CD4+in T-lymphocyte subset decreased and the ratio of CD4+/and CD8+was at its lowest when viral load reached its peak and then the ratio increased. As for the percent of CD8+,it increased during the whole acute period. The18-month-observation to the monkey model indicated that the percent of CD4+and the ratio of CD4+and CD8+exhibited fluctuating decrease trend as the infection progressed, whereas the viral load fluctuated around the set point by0.5logarithmic fluctuations. The variable related to TCM-syndrome like Cor exhibited fluctuating decrease trend as the infection progressed. The analysis of the data about the monkey model between AIDS period and death showed that the CD4+%, CD8+%, the value of CD4+/CD8+, and CD4+counts were closely related to the disease condition, but the decrease of T3was closely related to the deterioration of the disease, while the Cor level increased lto2months before death. The analysis of the survival condition of the animal model indicated that the median survival time of the monkey model infected with SIVmac239was about2years. The exploration of TCM pathogenesis through AIDS model showed that the cause of AIDS was Ai-pathogen, and the pathogenesis manifested as the invasion of Ai-pathogen, the struggle between the Ai-pathogen and healthy qi in acute infection period, then the incubation of Ai-pathogen and injury to the healthy qi in incubation period, then excess of Ai-pathogen and insufficiency of healthy qi in pre-AIDS period, and finally the deficiency of vitality in typical AIDS period.
4. The study created two composite model of Chinese rhesus monkey for AIDS with symptoms of spleen-deficiency and two composite model of Chinese rhesus monkey for AIDS with symptoms of kidney-deficiency successfully for the first time. That is spleen-deficiency type I, spleen-deficiency type II, kidney-deficiency type I, and kidney-deficiency type II. The clinical symptoms of spleen-deficiency composite model include decreased food intake, slow growth of the body mass, diarrhea and significant decline of the ratio of hematological indicators like triiodothyronine(T3), CD4+%and CD4+/CD8+,whereas in the kidney-deficiency composite model, the clinical symptoms showed as chills, listlessness, weight loss, diarrhea and significant decline of the ratio of hematological indicators like triiodothyronine (T3), Cor, CD4+%and CD4+/CD8+. Detailed research found that in spleen-deficiency type I, the effect of modeling factors on the disease of AIDS was not significant in the composite model creation stage, but significant after composite model creation stage, which will accelerate the progression of AIDS disease. It also turned out that the therapeutic effect of spleen-tonifying therapy was better than that of heat-releasing therapy for this model. As for spleen-deficiency type II, it showed similarity with the spleen-deficiency type I, but the deficiency of the spleen was more serious and the therapeutic effect of spleen-tonifying therapy was not better than that of heat-releasing therapy. The researches on kidney-deficiency type I indicated that the effect of modeling factors on the disease of AIDS was not significant in model creation stage, but significant on treatment and withdrawal period after the model creation period, which will accelerate the progression of AIDS disease.. The therapeutic effect of kidney-tonifying therapy was better than that of heat-releasing therapy, especially the effect of kidney-tonifying therapy8month after withdrawal. With regard to kidney-deficiency type II, it was much serious than kidney-deficiency type I, and the modeling factors had significant effect on AIDS disease during the model creation stage. Meanwhile, the therapeutic effect of kidney-tonifying therapy was better than that of heat-releasing therapy and the disease with this model progressed quickly without treatment. Based on the summary of experience of TCM intervention through animal AIDS model/composite model of Chinese rhesus monkey for AIDS symptoms, we optimized the original kidney-tonifying prescription and put forward the new one (optimized prescription) according to the pathogenesis of AIDS in monkey models.
5. The research results can be applied in several aspects. Based on the researches results of the pathogenesis of AIDS, the use of Shenfu injection for treating Chinese rhesus monkey model in the late stage successfully prolonged the survival time of the diseased monkey, alleviated the clinical symptoms and slowed down the progression of the disease. In addition, the results of treating monkey AIDS model with the optimized prescription of combination of single plant chemicals showed that the combination of TCM and single plant chemical E can significantly enhanced the efficacy of single plant chemical E without any obvious side effect. The experimental study on the effect of protease inhibitors indinavir on the plasma concentration in monkey AIDS animal model/healthy volunteers indicated that the combination of optimized prescription and indinavir significantly increased the plasma concentration of indinavir and played a role in slowing down the clearing rate of indinavir in the healthy volunteers.
Conclusions
1. The literature review shows that TCM has a significant effect on the prevention and treatment of AIDS and indicates that the in-depth research of TCM on AIDS prevention and treatment may make big progress or have a breakthrough. It also clarifies the fact that it is easier to use proper animal AIDS models to do AIDS research rather than human trial and the researches with animal models are controllable, reliable and repeatable procedures which can shorten the research process, reduce the confounding factors, and decrease the test risk, which proves the feasibility of the establishment of composite model for ADIS with TCM symptoms by Chinese rhesus monkey.
2. The establishment of standard procedures of empirical study on Chinese rhesus monkey model for AIDS (Standard Operating Procedure in the lab) basically guarantees accuracy and reliability of the research results and lays foundations to the standardization of operation, testing and observation of the experiment on Chinese rhesus monkey. The definition of normal fluctuating ranges of indicators of Chinese rhesus monkey under healthy condition provides related criteria to the correct judgment of the results.
3. Chinese rhesus monkey model for AIDS can reflect the changes of disease condition of AIDS, which is applicable in the various researches on AIDS. According to the Chinese rhesus monkey model, the cause of AIDS was Ai-pathogen, and the pathogenesis manifested as the invasion of Ai-pathogen, the struggle between the Ai-pathogen and healthy qi, then the incubation of Ai-pathogen and injury to the healthy qi, then excess of Ai-pathogen and insufficiency of healthy qi, and finally the deficiency of vitality and exhaustion of five zang and six fu (the internal organs), qi blood and ying and yang.
4. The composite AIDS model with TCM symptoms varies, and they have different effect on the progression of AIDS and TCM intervention. The study indicates that the disease condition of the composite AIDS model with TCM symptoms is more serious than the simple AIDS model. The comparison of the therapeutic effect indicates that the treatment with TCM is much better than non-treatment, among which kidney-tonifying therapy is better than heat-releasing therapy and the therapeutic effect of spleen-tonifying therapy and heat-releasing therapy is different in different models.
5. The application of the results proves the therapeutic effect of TCM on the treatment of AIDS, and also the benefits of treating AIDS with combined drug use of TCM and modern medicine, which theoretically provides some examples and research ideas for the reasonability of treating AIDS with the combined TCM of single plant chemicals and Western medicine.
引文
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