彝药“麻补”止血活性物质基础及机理研究

目的：阐明彝药“麻补”（狭叶重楼）的止血活性物质基础及机理。方法：采用植物化学技术从“麻补”药材中分离鉴定出一种C27甾体皂苷类化合物——重楼皂苷H（PSH）。测定了PSH 对正常小鼠的断尾出血时间（BT）、凝血酶原时间（PT）、活化的部分凝血活酶时间（APTT）、血浆纤维蛋白原（FIB）等凝血功能指标的影响。结果：PSH 可显著缩短BT,显示出明显的止血活性;PSH 对缩短正常小鼠PT、APTT 作用不明显,但可显著提高FIB。结论：重楼皂苷H 对内源凝血通路及外源凝血通路无明显促进作用,PSH 升高FIB 为其发挥止血活性的重要途径。     

Paris saponin I induces G<Subscript>2</Subscript>/M cell cycle arrest and apoptosis in human gastric carcinoma SGC7901 cells

The aim of this study was to investigate the effect of Paris saponinⅠ(PSⅠ)on human gas-tric carcinoma cell growth and apoptosis and to explore the potential mechanisms.The proliferation of SGC7901 cells was monitored by the MTT cell viability assay,while the nuclear morphology of apoptotic cells was assessed by Hoechst 33258 staining.Flow cytometry was performed to analyze the cell cycle progression of propidium iodide(PI)-stained SGC7901 cells and the apoptotic rate of annex-inⅤ/PI-stained cells.Western blotting was used to examine the expression of several cell cycle proteins,including cyclin B1 and Cdk1,and the apoptosis-regulated proteins Bcl-2,Bax,cytochrome c,procas-pase-9,and procaspase-3.The MTT assay demonstrated that PSⅠ could induce significant dose-and time-dependent inhibition of SGC7901 cell proliferation.Marked morphological changes,including condensation of chromatin,nuclear fragmentation and apoptotic bodies were clearly shown on Hoechst 33258 staining.PSⅠ treatment also resulted in the disruption of the cell cycle at G2/M and the induction of apoptosis.Following PSⅠ treatment,the cell cycle-related proteins cyclin B1 and Cdk1 were down-regulated.Expression of the pro-apoptotic protein Bax was increased,while anti-apoptotic protein Bcl-2 decreased.PSⅠ treatment resulted in elevated cytoplasmic cytochrome c and activation of the apoptotic proteases caspase-9 and caspase-3.These data indicate that PSⅠ acts as an inhibitor of proliferation in SGC7901 cells by inducing cell cycle arrest and mitochondria-dependent apoptosis.PSⅠ is a potential therapeutic agent against human gastric carcinoma.     

Survenue de cancers au cours de la sclérodermie systémique : facteurs de risque,impact sur la survie et revue de la littérature

IntroductionPatients with systemic sclerosis (SSc) have an increased risk of malignancy. In this study, we aimed to analyze the prevalence of cancer, the risk factors and the impact on overall survival.Patients and methodsWe analyzed clinical (history of cancer, toxic exposition, organ involvement), immunological and treatment data in a monocentric cohort of SSc patients followed between January 2004 and December 2017.ResultsTwo hundred and ten patients with SSc were included. During the follow-up, twenty-one patients (10 %) were diagnosed with malignancies. The underlying malignancies were breast adenocarcinoma (n = 6, 28%), lung cancer (n = 6, 28%), colorectal (colic adenocarcinoma, carcinoid tumor of the appendix), ovarian and cervix uteri, melanoma, kidney and papillary thyroid carcinoma (one of each). The median time between the first visit and the diagnosis of cancer was 4 [2–10] years. The overall survival in SSc patients with cancer was not significantly different from patients without cancer, with median survival during the first quartile (75%) at 12 years for patients with cancer and 11.6 years for those without cancer (P = 0.9). The history of renal scleroderma crisis (HR 10.99, IC95% [1.95–62.07]; P = 0.006) and the presence of anti-topoisomerase I antibodies (HR 5.5, IC95% [1.40–21.67]; P = 0.01) were associated with an increased risk of cancer, whereas the presence of gastroesophageal reflux was inversely associated with the cancer occurrence (HR 0.22, IC95% [0.056–0.867]; P = 0.03).ConclusionThe history of renal scleroderma crisis and the positivity of anti-topoisomerase I antibodies were associated with an increased risk of cancer in SSc patients in this monocentric study.     

加速溶剂萃取-HPLC-TOF/MS法同时测定重楼中6种甾体皂苷类成分

目的引入加速溶剂萃取(accelerated solvent extraction, ASE)技术,并结合HPLC-TOF/MS法同时测定重楼中6种甾体皂苷类成分的含量。方法加速溶剂萃取仪在120℃、1.17 MPa (1 700 psi)压力下,用70%的乙醇静态萃取样品6 min;6种重楼皂苷的含量用HPLC-TOF/MS同时测定,色谱柱: MGC18柱(3.0 mm×100 mm,3.0 μm);流动相: 乙腈与0.1%(V/V)甲酸水,梯度洗脱;流速: 0.8 ml/min;柱温: 25℃;进样量: 3 μl;离子源: ESI正离子模式;雾化气压力: 275.85 kPa(40 psi);干燥气流速: 10 L/min;干燥气温度: 350℃。结果重楼皂苷Ⅰ、重楼皂苷Ⅱ、重楼皂苷Ⅵ、重楼皂苷Ⅶ、薯蓣皂苷和纤细薯蓣皂苷的线性范围(μg/ml)分别为: 0.500 0~50.00(r=0.999 8)、0.422 3~42.23(r=0.999 7)、0.612 0~61.20(r=0.999 9)、0.714 0~71.40(r=0.999 5)、0.448 0~44.80(r=0.999 9)和0.436 0~43.60(r=0.999 7);各成分的平均回收率(n=6)分别为98.9%、98.0%、102.5%、101.9%、103.1%和97.9%。结论该法溶剂消耗少、耗时短、便于自动化,可实现高通量分析,可用于重楼中皂苷类成分的含量测定。     

滇重楼环阿屯醇合酶基因的克隆及序列分析

目的 获取滇重楼甾体皂苷合成途径关键酶环阿屯醇合酶基因(PpCAS)的全长cDNA序列,并进行序列分析.方法 利用同源克隆和RT-PCR技术获得PpCAS基因保守片段,采用RACE技术获得PpCAS基因的3'及5'末端序列,并采用生物信息学方法进行序列分析.结果 PpCAS基因全长cDNA为2 309 bp,其开放阅读框(ORF)为2 283 bp,可编码760个氨基酸的蛋白质;PpCAS推导的蛋白质相对分子质量为8.69×104,等电点(pI)为6.54;其氨基酸序列与GenBank中其他植物CAS的同源性在60％～83％ PpCAS蛋白.结论 从滇重楼中首次获得PpCAS基因cDNA全长序列,该基因具有CAS同源基因的典型特征.     

HPLC同时测定不同产地滇重楼中的6种重楼皂苷

目的对不同产地滇重楼的质量进行评价研究,摸清目前云南省境内的滇重楼资源情况,为滇重楼资源合理利用和保护提供科学依据。方法收集产自云南省境内13个州(市、地区)的滇重楼样品进行分析,采用HPLC方法测定其6种重楼皂苷Ⅰ,Ⅱ,Ⅵ,Ⅶ,PA,H。结果不同产地滇重楼皂苷含量差异较大。结论 重楼皂苷质量受产地、环境因素影响较大,滇重楼品质表现出一定的地域及生境依赖性。     

Biochemical response to ursodeoxycholic acid among PBC patients: a nationwide population-based study

Objective: To assess the proportion of PBC patients with a biochemical response to ursodeoxycholic acid (UDCA) in a population-based cohort and the association of biochemical response with outcomes.

Methods: All patients diagnosed with PBC in Iceland from 1991–2015 were identified. Patients taking UDCA for an adequate period of time were analyzed for treatment response according to the Barcelona, Paris I, Paris II and Toronto criteria and outcomes.

Results: Overall 182 females and 40 males were diagnosed with PBC and 135 patients were treated with UDCA. Overall 99 (73%) patients had adequate data on UDCA treatment and results of liver tests to assess biochemical response according to the Barcelona criteria, 95 (70%) according to the Toronto criterion and 85 (63%) according to the Paris I and II criteria. In all 74% (n?=?63), 67% (n?=?64), 54% (n?=?53) and 46% (n?=?39) responded to treatment according to the Paris I, Toronto, Barcelona and Paris II criteria. Among nonresponders according to the Paris I, Toronto, Paris II and Barcelona criteria, 50%, 39%, 33% and 30% developed cirrhosis versus 10%, 6%, 5% and 11% of responders, HR 5.36 (p?=?.002), 6.61 (p?=?.002), 10.94 (p?=?.003) and 2.21(p?=?.11), respectively. Age-adjusted mortality was significantly lower among responders according to the Paris I and Paris II criteria, HR 0.33 (p?=?.02) and 0.31 (p?=?.02), respectively.

Conclusion: Development of cirrhosis and higher mortality was significantly associated with a lack of biochemical response to UDCA. Frequent development of cirrhosis and increased mortality in nonresponders underlines the need for a more effective therapy than UDCA for this sizeable subgroup of patients.