The combination effects of Shen-Ling-Bai-Zhu on promoting apoptosis of transplanted H22 hepatocellular carcinoma in mice receiving chemotherapy

背景：肝细胞癌(Hepatocellularcarcinoma，HCC，简称肝癌)是我国常见恶性肿瘤之一，具有恶性程度高、生存时间较短、易转移复发、预后差等特点，死亡率在全世界所有肿瘤中高居第3位。我国肝癌高发，在肿瘤相关死亡中仅次于肺癌，居第2位。目前化疗(chemotherapy)是肿瘤治疗的常用手段，但同时易引起肝肾功能异常，导致骨髓抑制、消化道损伤，出现白细胞减少、恶心呕吐、食欲不振、腹痛腹泻、便秘等不良反应。西医尚缺乏具有良好疗效缓解化疗毒副反应的西药；中医临床诊疗实践发现，复方中药能够减轻患者化疗的毒副反应，具有增效减毒作用。大量临床观察证实健脾益气中药复方参苓白术散能改善肝癌患者...Background: Hepatocellular carcinoma (HCC) is one of the common malignant tumors in China, with a high degree of malignancy, short survival time, easy recurrence and poor prognosis. Hepatocellular carcinoma is currently the 3rd leading cause of cancer death in the world. While the mortality of hepatic carcinoma ranks second, just next to lung cancer, in China. At present, chemotherapy as an import...学位：医学硕士院系专业：医学院_中医内科学学号：2452014115357

Experimental Study on the Basic Effect of Huatan Anshen Decoction and on Orexin Expression in Hypothalamus of Insomnia Rats

目的：研究化痰安神方治疗失眠症的药效学基础及其对失眠大鼠下丘脑食欲素表达的影响,揭示本方部分药效学机制。方法：通过小鼠自发活动实验、戊巴比妥钠阈下及阈上睡眠协同实验探索化痰安神方治疗失眠症的药效学基础;通过对氯苯丙氨酸（PCPA）制造失眠大鼠模型,分为中药组、西药组、空白组、模型组,每组干预7天,使用免疫组化检测实验大鼠下丘脑食欲素（Orexin）。结果：（1）化痰安神方可抑制小鼠自发活动,与空白组比较,具有显著差异（P〈0.01）;（2）对戊巴比妥钠阈下入睡率的影响：中药组、对照组、西药组实验小鼠入睡率分别为：65%、72%和100%,西药组入睡率明显高于中药组、对照组（P〈0.01）;睡眠持续时间,中药组长于西药组（P〉0.05）,西药组长于对照组（P〈0.05）;（3）对戊巴比妥钠阈上睡眠情况的影响：中药组睡眠持续时间明显长于西药组（P〈0.01）,西药组长于对照组（P〈0.01）;（4）对氯苯丙氨酸（PCPA）致失眠大鼠各组下丘脑食欲素表达水平无差异（P〉0.05）。结论：化痰安神方具有一定的镇静催眠作用,可明显延长失眠患者的持续睡眠时间,其作用机理可能不是通过调节食欲素水平这一途径。Objective： To study the basic effect of Huatan Anshen decoction and the effect on orexin expressions in Hypothalamus of insomnia rats and reveal the partial pharmacodynamics mechanism. Methods： Spontaneous activity in mice and synergistic action test of pentobarbital sodium were observed to study the basic effect of Huatan Anshen decoction. The second part, through para chlorophenylalanine （PCPA） making rat model of insomnia, divided into Chinese medicine group and Western medicine group, blank group, model group, each group of rats were intervened for 7 days, Hypothalamus Orexin was detected by immunohistochemisty. Results： （1） Huatan Anshen Decoction can inhibit the spontaneous activity of mice, compared with the control group, with significant difference （P〈0.01）. （2） Effect on the rate of sleep pentobarbital sodium： Chinese medicine group and control group, western medicine group mice sleep rates were： 65%, 72% and 100%. The rate of western medicine group was significantly higher than that of traditional Chinese medicine group and control group （P〈0.01）. Sleep duration： Chinese medicine group was longer than the western medicine group （P〈0.05）, the western medicine group was longer than the control group （P〈0.05）. （3）Sleep duration of Chinese medicine group was significantly longer than the western medicine group （P〈0.01）, and the western medicine group was significantly longer than the control group（P〈0.01）. （4） There was no difference in the expression level of orexin （P〈0.05） in the hypothalamus of insomnia rats induced by phenylalanine （PCPA）. Conclusion： Huatan Anshen decoction has sedative and hypnotic effects, can prolong the duration of sleep in patients withinsomnia. The mechanism of Huatan Anshen decoction to prolong the duration of sleep in patients with insomnia may not be regulated by the level of orexin pathways.2013福建省中医药科研课题（批准号：wzln201311

Effects of Shenling Baizhu Powder on expression of apoptosis related proteins of tumor chemotherapy model mice with H22 hepatocellular carcinoma cells

目的:研究参苓白术散(SLBZP)对H22肝癌移植瘤小鼠化疗后肿瘤凋亡相关因子Caspase -3、; Caspase-9、X染色体连锁凋亡抑制因子(XIAP)、血小板源生长因子-BB(PDGF-BB)、血管生成素-1(Ang-1)、血管生成素-2; (Ang-2)表达的影响。方法:SPF级昆明小鼠64只,将肝癌H22细胞悬液(2*10~7/mL)接种于小鼠右前腋皮下,1周全部形成移植瘤后,行; 环磷酰胺(CTX,0.20g/kg,0.2mL/10g)腹腔注射,建立肿瘤化疗模型。随机分组为8组:模型组,CTX(0.02g/kg)组,SLB; ZP高、中、低(6.00、3.00、1.50g/kg)剂量组,SLBZP高、中、低剂量+CTX(0.02g/kg)组,给药14d后,运用蛋白印迹; 法(Western blot)测定小鼠瘤组织中Caspase-3、; Caspase-9、XIAP蛋白表达;酶联免疫吸附(ELISA)测定小鼠血清PDGF-BB、Ang-1、Ang-2表达。结果:SLBZP联合治疗; 各组抑瘤作用明显,CT X组,SLBZP高剂量组,SLBZP高、中剂量+C T X组抑瘤率分别为52.39%、; 45.84%、58.41%、52.77%。与模型组比较,各治疗组血清PDGF-BB、Ang-1、Ang-2及瘤组织中XIAP表达呈现出被下调的趋; 势,瘤组织中Caspase-3、Caspase-9表达呈现出被上调的趋势,且以SLBZP高剂量加CTX组变化幅度最为显著(P<0.01,P<0.; 05)。结论:SLBZP联合化疗可以更有效调节H22肝癌移植瘤小鼠肿瘤凋亡相关因子的表达,促进肿瘤细胞凋亡可能是其改善化疗的机制之一。Objective: To investigate the effects of Shenling Baizhu Powder (SLBZP); on expression of apoptosis related proteins such as Caspase-3,; Caspase-9, X-linked inhibitor of apoptosis (XIAP), PDGF-BB, Ang-1, Ang-2; of tumor chemotherapy model mice with H22 hepatocellular carcinoma; cells. Methods: Sixty-four Kunming mice were subcutaneously injected; with suspension of H22 hepatocellular carcinoma cells (2*10~7/mL) into; the right anterior armpit. After 1 week, all transplanted tumors were; formed and the mice were received intraperitoneal injection with cytoxan; (CTX) with the dosage of 0.2g/kg to establish the tumor chemotherapy; model. Then mice were randomly divided into eight groups. CTX group was; treated with CTX (0.02g/kg), the model group was treated with; physiologic saline, three SLBZP groups were treated with SLBZP (6.00,; 3.00, 1.50g/kg), and other three groups were treated with SLBZP (6.00,; 3.00, 1.50g/kg) plus CTX (0.02g/kg). After all groups were treated for; 14 days, Western blot was used to detect the expression of Caspase-3,; Caspase-9 and XIAP proteins in tumor tissue, and protein expression of; PDGF-BB, Ang-1, Ang-2 in blood was measured by enzyme-linked; immunosorbent assay (ELISA). Results: The combined treatment group had; obvious effect on tumor inhibition. Tumor inhibitory ratios in CTX; alone, SLBZP (H), SLBZP (H) plus CTX, and SLBZP (M) plus CTX groups were; 52.39%, 45.84%, 58.41% and 52.77% respectively. Compared to the model; group, the protein expressions of PDGF-BB, Ang-1, Ang-2, and XIAP in; tumor tissue were obviously lower than those in all treated groups. And; the expressions of Caspase-3 and Caspase-9 in tumor tissue showed a; trend of up-regulation. And the effect in SLBZP (H) plus CTX group was; the most significant (P<0.01, P<0.05). Conclusion: SLBZP can more; effectively adjust the expression of apoptosis related proteins of tumor; in mice with H22 hepatocellular carcinoma cells, and promoting tumor; apoptosis may be one of the possible mechanisms of SLBZP to improve; chemotherapy in treating hepatic cancer.国家自然科学青年基金项目; 福建省自然科学基金项目; 福建省大学生创新创业训练项

Effects of Taohong Siwu Decoction on the expression of pathological angiogenesis correlation factors including VEGF,KDR and Flt-1 in hepatic fibrosis mice

目的:从整体水平研究复方桃红四物汤对四氯化碳所致肝纤维化模型小鼠肝组织血管内皮生长因子(VEGF)、含激酶插入区受体(KDR)及Flt-1蛋白与mRNA表达的影响。方法:昆明小鼠60只,随机分组为空白组,模型组,秋水仙碱组,桃红四物汤高、中、低(17.00、8.50、4.25g/kg)剂量组,采用左侧腹股沟处皮下注射四氯化碳花生油溶液建立小鼠肝纤维化模型,给药6周后,运用酶联免疫吸附测定(ELISA)法检测肝组织VEGF蛋白含量;蛋白印迹法(Western blot)测定小鼠肝脏KDR、Flt-1蛋白的表达;逆转录PCR(RT-PCR)法测定小鼠肝脏VEGF、KDR及Flt-1 mRNA表达。结果:桃红四物汤治疗后,桃红四物汤高、中剂量组肝组织VEGF蛋白表达量较模型组降低(P<0.05,P<0.01),中药高剂量组肝组织KDR与Flt-1蛋白表达较模型组显著降低(P<0.05);而桃红四物汤高、中剂量组肝组织VEGF、KDR与Flt-1 mRNA表达较模型组、秋水仙碱组明显减少(P<0.01),并且呈现一定的量效正相关。结论:桃红四物汤能够降低肝纤维化病理血管生成相关因子的表达,减缓病理性的微血管生成可能是其改善肝纤维化的机制之一。Objective:To investigate the effects of Taohong Siwu Decoction(THSWD) on protein and mRNA expression of vascular endothelial growth factor(VEGF),kinase insert domain containing receptor(KDR) and Flt-1 in hepatic fibrosis model mice caused by CCI_4.Methods:Sixty Kunming mice were randomly divided into blank,model,colchicine(control)and three THSWD(17.00,8.50,4.25g/kg) groups,with 10 mice in each group.Except the blank group,the other groups' mice were subcutaneously injected with carbon tetrachloride(CCI_4)-arachis oil solution in left inguen to establish hepatic fibrosis mouse model.After administration for six weeks,the content of VEGF protein in liver tissue was measured by enzyme-linked immunosorbent assay(ELISA),protein expressions of KDR and Flt-1 in liver tissue were detected by Western blotting,and mRNA expressions of VEGF,KDR and Flt-1 in liver tissue were measured by reverse transcription polymerase chain reaction(RT-PCR).Results:After treatment,the content of VEGF protein in THSWD(17.00,8.50g/kg) groups were lower than that in model group(P<0.05,P<0.01).The protein expressions of liver tissue KDR and Flt-1 in THSWD(17.00g/kg) group were less than that in model group(P<0.05).The mRNA expressions of liver tissue VEGF,KDR and Flt-1 were significantly lower than that in both model and colchicine groups(P<0.01),which showed a certain positive correlation between dose and effect.Conclusion:Taohong Siwu Decoction could decrease the expression of pathological angiogenesis correlation factors of hepatic fibrosis,thus helpful to induce the formation of pathological capillary vessels,which may be one of the possible mechanisms of ameliorating hepatic fibrosis.国家自然科学青年基金项目(No.81202659);; 福建省自然科学基金项目(No.2014J01373);; 厦门大学新聘教师科研启动基金项目(No.ZK1014);; 福建省大学生创新创业训练项目(No.DC2013174)~

抑郁症案

患者,女,25岁。就诊日期:2014年9月2日。主诉:入睡困难、情志淡漠1年,加重1个月。病史:1年前因工作压力大,逐渐出现入睡困难,睡眠较浅,夜间易醒,醒后不易入睡,每晚睡眠时间约1H,伴情志淡漠、寡言少语、情绪低落、心烦易怒、兴趣减退、悲观、思维迟钝。于当地医院诊断为抑郁症,予口服阿米替林、多虑平等多种抗抑郁药,症状有所缓解,但因药物不良反应较大,患者间断停药致病情

Quick identfication for pathogenic vibrio of aquacultivated creatures by UPPCR-SSCP

采用通用引物PCR配合SSCP分析即UPPCR SSCP技术 ,对弧菌科弧菌属中的副溶血弧菌、溶藻酸弧菌、费尼斯弧菌、鳗弧菌、哈维氏弧菌的 1 6S RNA基因进行检测。结果发现 ,UPPCR SSCP技术能较好地鉴别细菌的不同种 ,但对同种不同株细菌的区分较困难。认为PCR SSCP是将病原菌判定至种的有力工具。此研究对水产养殖病原菌的快速诊断与及时防治有重要意义。By universal primer PCR (UPPCR) amplification and SSCP, 16S DNA gene fragments of Vibrio parahaemolyticus, Vibrio alginolyticus, Vibrio furnissii, Vibrio anguillarum, Vibrio harveyi had been detected in this paper. It was found that different species of bacteria could been easily identified by this technique, but the same species could not. The findings suggest that UPPCR SSCP may be a potential tool for identifying the species of pathogenic bacteria. Therefore, this study will be of benefit in quick diagnosis and precaution in aquacultivated creatures.国家自然科学基金!(39770 5 85号 );; 国家海洋局第三海洋研究所海洋生物工程重点实验室开放课题;; IFS基金资助项 目!(A/ 2 338 2号