慢性乙型肝炎和肝硬化“异病同证”的全基因组甲基化分析

目的：本文从全基因组DNA甲基化的角度探讨慢性乙型肝炎（慢乙肝）和慢乙肝后肝硬化（肝硬化）湿热内蕴证、肝郁脾虚证和肝肾阴虚证异病同证的生物学基础。方法：采取慢乙肝及肝硬化湿热内蕴证、肝郁脾虚证和肝肾阴虚证患者和健康志愿者外周血样本,提取DNA进行HumanMethylation450K芯片检测及分析。结果：慢乙肝和肝硬化异病同证湿热内蕴证特异性差异甲基化位点有9个,覆盖9个基因;肝郁脾虚证有30个,覆盖20个基因;肝肾阴虚证有22个,覆盖14个基因。其中,与正常组相比,湿热内蕴证中KCTD2和NAV1,肝郁脾虚证中LGR6和SH2D4B及肝肾阴虚证中CYP2E1、PCSK6、DEXI、HIST1H3B和SULT1C2的差异位点甲基化程度变化较大（|Delta Beta|>0.15）。结论：慢乙肝与肝硬化基因甲基化可能与其湿热内蕴证、肝郁脾虚证和肝肾阴虚证的形成有关。     

Effects of different directions of moxibustion therapy on hemorheology in rats with blood stasis due to cold retention

Objective

To investigate the effects of different directions of moxibustion therapy on hemorheology in rat models with blood stasis due to cold retention.

Methods

A total of 32 Wistar rats were randomly divided into 4 groups, based on the random digits table, including a normal group, a model group, a moxibustion along the meridian group, and a moxibustion against the meridian group, with 8 rats in each group. Except the normal group, the other 3 groups were used to make the rat models with blood stasis due to cold retention. Rats in the moxibustion along and against the meridian groups accepted moxibustion therapy in different directions on the trunk segment of the Bladder Meridian after successful modeling. Moxibustion for 10 min every day, and 7 d as a course of treatment. Two courses of treatment (a total of 14 d) were carried out. Quantitative score of signs and symptoms change was observed once a week, with a total of 7 times. Hemorheological indexes of rats were detected when the treatment finished.

Results

The quantitative scores of signs and symptoms in the model, moxibustion along the meridian and against the meridian groups were significantly higher than those in the normal group after modeling (P<0.05). When the treatment finished, the quantitative score of signs and symptoms in the model group was significantly higher than that in the normal group (P<0.05); the quantitative scores of signs and symptoms of rats in the moxibustion along and against the meridian groups were significantly lower than that in the model group (P<0.05); the quantitative score of signs and symptoms in the moxibustion along the meridian group was significantly lower than that in the moxibustion against the meridian group (P<0.05); the whole blood viscosity and erythrocyte aggregation index in the model group were significantly higher than those in the normal group (P<0.05), however the increases of erythrocyte hematocrit and erythrocyte sedimentation rate were not obvious (P>0.05); whole blood viscosity and erythrocyte aggregation index in the moxibustion along and against the meridian groups were significantly lower than those in the model group (P<0.05), while there were no statistically significant differences between the moxibustion along the meridian group and moxibustion against the meridian group (P>0.05).

Conclusion

Moxibustion along and against the meridian both showed different degrees of improvement for hemorrheology and symptoms of blood stasis due to cold retention. Improvement for symptoms of blood stasis due to cold retention was better in the moxibustion along the meridian group than that in the moxibustion against the meridian group. The influence on rat hemorrheology showed no significant difference between moxibustion along the meridian and moxibustion against the meridian.

     

肝癌和大肠癌“异病同证”的证候分布及其与临床实验室指标的关联研究＊

目的 调查原发性肝癌（肝癌）与大肠癌患者的中医证候分布，探讨血常规、肝肾功能、免疫功能和肿瘤指标等临床实验室指标与中医证候的关联。方法 运用《中医肝癌与大肠癌病例汇报表（CRF表）》采集中医四诊信息、客观指标等，分析肝癌与大肠癌的证型分布及各证型在临床客观指标上的表现特点及“异病同证”的规律。结果 460例肝癌和759例大肠癌患者具有肝肾阴虚、脾虚、瘀血、湿热、实热、气虚、阴虚、隐证8种单证及各种兼证，其中脾虚证（27%）、肝肾阴虚证（12%）、隐证（11%）、湿热证（9%）、肝肾阴虚兼脾虚证（8%）为肝癌和大肠癌临床较为常见的五种中医证型。血常规HB、肝肾功能TBIL、ALB和GGT在五种证型间整体分布有显著差异（P < 0.05），AFP在肝癌证型间差异无统计学意义（P > 0.05），在大肠癌证型间差异显著（P < 0.001）。但免疫功能和细胞因子在上述五种证型间整体分布差异无统计学意义（P > 0.05）。结论 脾虚证、肝肾阴虚证、肝肾阴虚兼脾虚证、隐证、湿热证是肝癌与大肠癌证候分布的前五位，其证候分布与HB、TBIL、ALB、GGT和AFP存在关联，提示这些临床实验室指标可能作为肝癌和大肠癌“异病同证”辩证的参考依据。     

基于网络药理学方法的左金丸治疗结直肠癌的作用机制分析

目的：运用网络药理学方法探究左金丸治疗结直肠癌的潜在效应机制。方法：依托中药系统药理学分析平台收集左金丸中黄连和吴茱萸的化学成分和靶点信息,通过疾病基因库收集结直肠癌疾病基因,并通过维恩分析、蛋白互作分析以及Cytoscape的可视化功能,构建化合物-靶点网络、蛋白互作(PPI)网络和成分-靶点-通路网络,分析左金丸治疗结直肠癌的效应机制。结果:从左金丸化合物-靶点网络得到234种成分和482个核心靶点(其中活性成分36个,核心靶点72个),结直肠癌基因737个,PPI网络包含50个蛋白,其主要蛋白涉及PIK3CA、AKT1、TP53、HIF-1α、BAX和MYC等。基因GO本体条目283条,涉及核质、细胞凋亡、基因表达调控等方面, KEGG通路58条,主要涉及有癌症的途径、癌症中的蛋白多糖、结肠癌、膀胱癌、乙型肝炎,以及HIF-1信号通路、PI3K信号通路和FOXO信号通路等,其结果有待进一步的实验验证。结论：结合文献报道和预测分析,左金丸可能的作用机制与以下相关：①调节机体免疫耐受能力,增强宿主免疫抵抗,抑制炎症对肠粘膜的持续性刺激,进而阻止了细胞的异常增生;②通过上调抑癌基因,下调癌症相关因子,促进肿瘤细胞发生凋亡;③降低肿瘤细胞的耐受缺氧环境,促进细胞细胞发生凋亡及上皮间质的转移。     

大肠癌和肝癌术后“异病同证”的代谢组学研究

目的：本文主要探讨大肠癌和肝癌术后肝肾阴虚证、脾虚证和湿热证“异病同证”的血浆共同代谢物及其代谢机制。方法：对大肠癌和肝癌术后肝肾阴虚证、脾虚证、湿热证和隐证的血浆代谢物谱进行代谢组学（GC-MS）检测,采用主成分分析（PCA）方法寻找其“异病同证”的共同代谢物,并通过KEGG数据库分析代谢通路。结果：大肠癌和肝癌术后肝肾阴虚证共同的代谢物是甘氨酸,尿素,色氨酸和丙氨酸,其代谢调控以氨基酸的降解为主;脾虚证共同的代谢物是甘氨酸、尿素、色氨酸、葡萄糖、赖氨酸和肌醇,氨基酸的降解和糖类的分解功能受到影响;湿热证的共同代谢物是甘氨酸、尿素、色氨酸、葡萄糖、丙酸、甘露醇、山梨醇和赖氨酸,以影响糖类物质分解和供能过程为主要特征。结论：代谢组学方法是研究“异病同证”的有效工具之一。大肠癌、肝癌术后在肝肾阴虚证、脾虚证和湿热证各有其共同的代谢物以及代谢机制。