CCL_4诱导大鼠肝损伤模型的代谢组学及茵陈蒿汤的干预作用研究

基于代谢组学的系统化理论,以UPLC/QTOF分析系统为核心手段,结合生物样品分析制备方法,研究CCL_4诱导肝损伤大鼠的代谢组变化,以及茵陈蒿汤干预后的肝损伤大鼠代谢组的回调趋势。同时,借助MS解析理论和主成分分析软件,明晰表征大鼠肝损伤的5个内源性特征生物标记物,并结合系统生物学的理论还原肝损伤的机理与实质,以及茵陈蒿汤的肝损伤防治机理,从药物代谢组学角度对经典方剂防治肝损伤给出全新解释。     

中药及其复方的体内代谢研究现状与未来发展

中药及复方的体内代谢研究是当今探讨体内过程和阐明药效物质基础及作用机制的前沿热点领域，也为深层诠释中医药科学内涵、中医药理论的现代创新以及中药制药产业链的现代化提供了有效的途径。中药及其复方体内代谢研究有助于揭示药效物质基础、药效成分的体内动态规律，明晰体内药效成分与表观药效间的时量与时效间的关联，破译合“诸药之长”以成“有制之师”背后隐藏的配伍规律科学真谛，为打破长久以来困扰中医药“知其然，不知其所以然”的尴尬局面提供契机。对于建设符合中医药学科规律的ADME／Tox新药筛选平台，指导脱胎于中医经方又高于经方的创新型中药制剂的开发奠定了基础。本文对近年来中药及其复方的体内代谢研究现状和未来发展进行了综述和评价。     

Lifespan extension by n-butanol extract from seed of Platycladus orientalis in Caenorhabditis elegans

Aim of the study

As a traditional Chinese medicine, seed of Platycladus orientalis(Linnaeus) Franco has been extensively used as a tonic and sedative remedy. The present study was conducted to investigate whether lifespan was extended and the mechanisms of n-butanol extract from seed of Platycladus orientalis (BSPO) in Caenorhabditis elegans. The findings could provide the pharmacological basis for a treatment in traditional medicine.

Materials and methods

Lifespan extension by BSPO was evaluated under normal culture conditions and in a stress test. A possible mechanism of the anti-aging effect of BSPO, a change in the stress-resistance of related proteins, was also investigated in C. elegans.

Results

It has been shown that BSPO could significantly extend lifespan of C. elegans in a concentration dependent manner under normal culture conditions and stress. Further studies demonstrated that BSPO treatment significantly decreased reactive oxygen species (ROS) accumulation, up-regulated resistance to stress of related proteins, including glutathione S-transferase-4 (GST-4) and heat shock protein-16.2 (HSP-16.2), and reduced the amount of lipofuscin in transgenic C. elegans.

Conclusion

These results indicated that BSPO extended the lifespan, which could be attributed to its direct ROS scavenging activity, reducing the amount of lipofuscin and increasing the expression of gens associated with resistance to stress. These obtained data provided valuable support for traditional clinical practice to extend lifespan and to provide tonic remedy.     

Identifying dysfunctional miRNA-mRNA regulatory modules by inverse activation,cofunction, and high interconnection of target genes: A case study of glioblastoma

Background

Accumulating evidence demonstrates that complex diseases may arise from cooperative effects of multiple dysfunctional miRNAs. Thus, identifying abnormal functions cooperatively regulated by multiple miRNAs is useful for understanding the pathogenesis of complex diseases.

Methods

In this study, we proposed a multistep method to identify dysfunctional miRNA-mRNA regulatory modules (dMiMRMs) in a specific disease, in which a group of miRNAs cooperatively regulate a group of target genes involved in a specific function. We identified dysfunctional miRNAs, which were differentially expressed and inversely regulated most of their target genes, by integrating paired miRNA and mRNA expression profiles and miRNA target information. Then, we identified cooperative functional units, in each of which a pair of miRNAs cooperatively repressed function-enriched and highly interconnected target genes. Finally, the cooperative functional units were assembled into dMiMRMs.

Results

We applied our method to glioblastoma (GBM) and identified GBM-associated dMiMRMs at the population, subtype, and individual levels. We identified 5 common dMiMRMs using all GBM samples, 3 of which were associated with the prognosis in patients with GBM and were better predictors of prognosis than were miRNAs or mRNAs alone. By applying our approach to GBM subtypes, we found consistent dMiMRMs across GBM subtypes, and some subtype-specific dMiMRMs were observed. Furthermore, personalized dMiMRMs were identified, suggesting significant individual differences in different patients with GBM.

Conclusions

Our method provides the capability to identify miRNA-mediated dysfunctional mechanisms underlying complex diseases.     

中性粒细胞异常增殖相关的BCR/ABL-慢性白血病的临床特征

目的:探讨与中性粒细胞异常增殖相关的BCR/ABL-慢性白血病的临床特征.方法:报告3例不同类型与中性粒细胞异常增殖相关的BCR/ABL-慢性白血病并结合文献进行讨论.结果:3例均为老年男性,表现为以中性粒细胞为主的白细胞异常增多、BCR/ABL融合基因阴性.例1诊断为慢性中性粒细胞白血病(CNL),脾大,外周血以成熟中性粒细胞增高为特征,胞浆内中毒颗粒易见,无明显病态造血,中性粒细胞碱性磷酸酶(NAP)染色强阳性,骨髓粒系增生明显活跃.例2诊断为不典型慢性髓细胞白血病(aCML),血涂片示不成熟粒细胞比例增高伴粒系病态造血,尤其核染色质异常浓聚更为明显,骨髓象表现为髓系增生和病态造血,原始细胞轻度增多.例3诊断为慢性粒单核细胞白血病(CMML),外周血成熟单核细胞数量和比例异常偏高,粒系病态如核分叶过多、环状核和假性Pelger-Huet畸形等易见,骨髓髓系增生明显活跃伴病态造血,无原始细胞增多.结论:中性粒细胞异常增殖相关的BCR/ABL-慢性白血病较为少见,易于误诊,细胞形态学特征是诊断BCR/ABL-慢性白血病的基础.临床表现、多种实验室检查结果尤其是分子生物学等有助于正确诊断这类疾病.     

术前血清CA125、HE4和中性粒细胞与淋巴细胞比值联合检测在子宫内膜癌诊断中的作用

目的:探讨血清糖类抗原125（carbohydrate antigen 125,CA125）、人附睾蛋白4（human epididymis protein 4,HE4）、中性粒细胞与淋巴细胞比值（neutrophil-lymphocyte ratio,NLR）联合检测在子宫内膜癌诊断中的作用。方法:选取42例子宫内膜癌患者、50例子宫内膜良性疾病患者和50例健康体检人群。采用SYSMEX XN550全自动血液分析仪计数术前外周血中性粒细胞和淋巴细胞,计算NLR;采用Maglumi4000全自动化学发光仪检测术前血清CA125、HE4水平。采用ROC曲线分析CA125、HE4、NLR和三者联合指标在诊断子宫内膜癌中的作用。结果:外周血NLR在健康组、良性组和子宫内膜癌组中逐渐增高,且差异有统计学意义（P<0.05）;子宫内膜癌组HE4表达量显著高于良性组与健康组（P<0.05）,而良性组与健康组之间差异无统计学意义（P>0.05）;CA125表达量在三组中差异无统计学意义（P>0.05）。CA125、HE4、NLR及三者联合标记物的AUC分别为0.530、0.733、0.795、0.823,当分别取它们的临界值时,特异性分别为70.8%、85.1%、61.7%、83.0%,敏感性分别为40.0%、63.3%、86.7%、73.3%。单项指标NLR的敏感性最高,HE4的特异性最高,联合指标的特异性和敏感性都很高。结论:术前血清CA125、HE4和NLR联合检测具有较高的特异性和敏感性,联合检测可以互为补充,提高子宫内膜癌的诊断准确率,对子宫内膜癌的诊断具有指导意义。     

IFN-γ通过诱导上皮间质转化增强人乳头状甲状腺癌细胞株的侵袭转移能力

目的:观察IFN-γ对人乳头状甲状腺癌细胞株TPC-1侵袭和转移能力的影响,并探讨上皮间质转化过程（EMT）在其中的作用。方法:采用划痕愈合实验和Transwell侵袭实验检测IFN-γ对TPC-1侵袭转移能力的影响;实时定量PCR和Western检测EMT标志物E-cadherin,N-cadherin,Vimentin的mRNA水平及蛋白水平变化;免疫荧光检测上述标志物的定位表达。结果:划痕愈合实验和Transwell侵袭实验结果显示:IFN-γ增强了TPC-1的侵袭转移能力（P＜0.05）,实时定量PCR结果显示EMT的上皮标志物E-cadherin的mRNA表达水平明显下降,间质标志物Vimentin的mRNA表达上调,而N-cadherin的mRNA未见明显变化。Western结果显示E-cadherin在IFN-γ作用下表达下调;N-cadherin与Vimenin表达上升;免疫荧光结果与Western结果一致。结论:INF-γ可以通过诱导EMT,促进TPC-1侵袭转移,提示炎性介质IFN-γ可能参与人乳头状甲状腺癌的侵袭转移。     

紧密连接蛋白Claudin-3与三阴性乳腺癌患者临床特征的相关性

目的:探讨紧密连接蛋白Claudin-3与三阴性乳腺癌患者临床特征的相关性。方法:自2012年3月至2017年3月,收集我院收治的三阴性乳腺癌患者108例,行乳腺癌根治术,术中取乳腺癌标本和癌旁标本,检测组织中紧密连接蛋白Claudin-3表达情况,分析紧密连接蛋白Claudin-3与患者临床病理参数的相关性。对患者随访1年,分析术后1年复发患者与不复发患者组织中紧密连接蛋白Claudin-3的差异。结果:乳腺癌组织和癌旁组织中紧密连接蛋白Claudin-3表达无统计学差异(P=0.331)。与紧密连接蛋白Claudin-3阴性的患者相比,紧密连接蛋白Claudin-3阳性的患者TNM分期为Ⅲ期的比例显著增高(52.46% vs 29.79%,P=0.018),淋巴结转移比例增高(55.74% vs 29.79%,P=0.007),术后复发率显著增高(26.23% vs 10.64%,P=0.042)。结论:紧密连接蛋白Claudin-3与淋巴结转移和TNM分期有关,对预测患者术后复发具有一定价值。     

miR-193a-3p在食管癌细胞放射抵抗作用的初步研究

目的 探讨miR-193a-3p在食管鳞癌放射耐受性机制中的作用。方法 通过6 MV X射线对4个食管癌细胞系进行照射,采用MTT法检测出相对敏感系及耐受系细胞;茎环引物实时定量PCR法检测miR-193a-3p、miR-155、miR-22-3p在2个细胞中的表达,miR-193a-3p作为表达差异较为明显的microRNA被挑选进行下一步研究。分别合成并转染miR-193a-3p的mimic (3PM)或 antagomiR (3PA)序列及小干扰RNA (si-LOXL4)以提高或抑制其在细胞中的表达水平,MTT法和流式细胞分析术检测miR-193a-3p及其下游基因LOXL4对于放射敏感性的影响。结果 筛选出相对放射敏感细胞系(KYSE510)及耐受细胞系(KYSE410);miR-193a-3p在两系细胞中的表达水平差异显著高于miR-155、miR-22-3p (1.00:21.13);KYSE510细胞中转染mimic提高其表达后,与对照组相比,其放射敏感性降低,细胞凋亡比例显著下降11.10%(P<0.05),而KYSE410细胞中转染antagomiR后,其敏感性增加(P<0.05)。作为miR-193a-3p的下游基因,LOXL4的表达抑制也受到miR-193a-3p的调控,转染si-LOXL4降低其表达,与对照组相比放射敏感性也降低,细胞凋亡比例下降7.07%(P<0.05)。结论 miR-193a-3p可能通过调控基因LOXL4促进食管癌细胞放射耐受性。