Antifungal alkaloids from the fresh rattan stem of Fibraurea recisa Pierre.

Aim of the study

The rattan stem of Fibraurea recisa Pierre. is known as an ethno-remedy commonly used for the treatment of various skin diseases by the minority Yao, Zhuang and Miao in Yunnan Province of China. The present study was designated to evaluate its antifungal activity, and to root out the antifungal substances from this ethical herb.

Materials and methods

The in vitro antifungal assay was performed by agar diffusion test for extracts and fractions. Then, the active fractions were submitted to column chromatography on silica gel and LH-20 to isolate their compounds. And the antifungal activity of pure compounds has been examined by checkerboard microdilution test. Nine Candida strains and one Cryptococcus strain were used for the bioassay.

Results

The MeOH extract exhibited significant antifungal activity, and the alkaloidal fractions were deduced as main active component. Subsequent studies led to the identification of a new alkaloid fibrecisine (1) and 21 known alkaloids including berberines, tetrahydroberberines and aporphine derivatives. The bioassay result indicated that the berberines showed more potent activity than aporphine derivatives against the test Candida strains, while tetrahydroberberines showed very weak activity against Cryptococcus neoformans.

Conclusion

The new alkaloid fibrecisine (1) was identified as 1,2-methylenedioxy-8-hydroxy-6a(R)-aporphine by detailed spectral analysis. The rattan stem of Fibraurea recisa Pierre. is an effective antifungal herb, and its major active component is alkaloidal compounds. Bioassay tests revealed that the water-soluble berberines are the most important antifungal substances. The study provides preliminary scientific validation for the traditional medicinal use of this ethno-remedy.     

吉西他滨固定剂量率输注联合奥沙利铂一线治疗晚期胰腺癌的Meta分析

背景与目的:有研究提示吉西他滨(Gemcitabine,GEM)固定剂量率(Fixed-dose rate,FDR)输注治疗晚期胰腺癌似有较好的疗效,Meta分析也显示含铂类联合化疗优于GEM单药化疗,本文试图通过Meta分析,探讨GEM FDR输注联合奥沙利铂(GEMOX)一线治疗晚期胰腺癌的地位和价值.方法:通过MEDLINE、EMBASE、ASCO等数据库及论文集检索国内外的相关文献.选择治疗组为GEMOX方案化疗,对照组为标准GEM单药化疗的晚期胰腺癌随机对照试验.由2位评价者分别按上述检索策略收集资料,按纳入标准入选,主要对总生存率及主要不良反应进行Meta分析.结果:从182篇文献中筛选出符合纳入标准的2个随机对照试验,共涉及869例患者.与GEM单药组比较,GEMOX组半年生存率提高9%(95%CI 0.03～0.16,P=0.005),1年生存率提高5%(95%CI-0.01～0.11,P=0.08),客观有效率提高6%(95%CI 0.02～0.10,P=0.006);WHO Ⅲ/Ⅳ度贫血发生率下降5%(95%CI-0.08～-0.01,P=0.01),恶心/呕吐提高13%(95%CI 0.08～0.18,P＜0.001),神经毒性增加14%(95%CI 0.04～0.24,P=0.009),粒细胞减少症、血小板减少症两组相似,差异无统计学意义.结论:现有的证据提示,GEM固定剂量率输注联合奥沙利铂组成的GEMOX方案一线治疗晚期胰腺癌可能有较好的应用前景,值得进行进一步的临床试验.     

NF-κB介导的EBV-LMP1在Rat-1细胞转化和成瘤中的作用

背景与目的:EB病毒潜伏性膜蛋白1(latent membrane protein 1,LMP1)是病毒基因组编码的致瘤蛋白.本研究拟探讨LMP1在细胞转化和致瘤过程中的作用机制.方法:采用基因重组方法构建LMP1和显性负突变IкBα逆转录病毒质粒pLNSX-LMP1和pBabe-IкBα,分别与含有转录因子NF-кB启动子的荧光素酶表达质粒共转染293细胞,单光子检测仪测定LMP1对NF-кB的活化作用及IкBα对NF-кB的抑制作用;同时将2种重组病毒载体分别导入PA317细胞包装,获取2种逆转录病毒(retrovirus,RV),单独(RV-LMP1)或先后(先RV-LMP1后RV-IкBα)感染体外培养的Rat1细胞,检测转导细胞的集落形成能力及裸鼠成瘤能力.结果:当pBabe-IкBα与pLNSX-LMP1以1∶1共转染,IкBα能使LMP1活化NF-кB的作用降低75%;当以3∶1共转染,LMP1的活化作用几乎全部被抑制.IкBα能明显抑制RV-LMP1感染细胞的恶性表型:平皿克隆形成数从368±7和287±17分别降至59±6和8±2(n=3,P＜0.001);软琼脂集落形成数从477±13和347±10分别降至61±15和95±7(n=3,P＜0.001);裸鼠成瘤能力显著降低,瘤体积自(1.61 0.23)cm3降至(0.20±0.08)cm3(n=5,P＜0.001).结论:EB病毒LMP1促Rat1细胞恶性转化作用主要依赖NF-кB的活化.     

鼠双微体2与核糖体蛋白L23在胃癌组织中的表达及临床意义

目的:研究胃癌中鼠双微体2(murine double mimute 2,MDM2)与核糖体蛋白L23(ribosomal protein L23,RPL23)的表达及其临床相关性,初步探讨他们在胃癌发生发展中的生物学意义.方法:采用免疫组织化学染色方法检测90例胃癌组织和30例胃癌旁正常组织中MDM2和RPL23蛋白的表达;统计学分析他们表达的相关性及其与临床病理因素、患者生存时间的联系.结果:胃癌组织中MDM2表达阳性率与癌旁对照组比较显著性增高(62.2%vs 40.0%,P0.05),而RPL23蛋白表达显著性降低(30.0%vs 63.3%,P0.05).MDM2和RPL23在胃癌中的表达呈负相关(r=-0.23,P=0.029).多因素分析显示MDM2高表达和RPL23低表达、淋巴结转移、肿瘤入侵深度、肿瘤的大小对胃癌患者生存预后的影响有统计学意义.结论:MDM2、RPL23的表达与胃癌的发生发展具有一定的相关性,针对两者的信号通路或许可作为胃癌预后的标志和新型治疗靶标.     

吉西他滨联合化疗一线治疗晚期胰腺癌生存结果的亚组meta分析

 【目的】Meta分析提示吉西他滨（GEM）联合化疗一线治疗晚期胰腺癌优于标准的GEM单药化疗,在此基础上进行亚组生存结果的meta分析及资料更新,旨在寻找确切有效的化疗方案。【方法】通过MEDLINE、EMBASE、ASCO、ECCO等数据库及论文集检索相关文献。按纳入标准筛选新增文献并进行资料更新。主要对各亚组进行半年生存率、其次是1年生存率的meta分析。【结果】17个随机对照临床试验（RCT）共3 821例患者纳入分析,按化疗方案分为GEM联合顺铂（GEMDDP）、GEM固定剂量率输注联合奥沙利铂（GEMOX）、联合5FU（GEMFU）、联合卡培他滨（GEMCAP）以及联合伊立替康（GEMIRI）等5个亚组,各亚组半年生存率的治疗优势（RD）分别为5%（P=0.24）、9%（P=0.005）、2%（P=0.46）、7%（P=0.03）和-1%（P=0.88）;1年生存率RD为6%（P=0.11）、5%（P=0.07）、4%（P=0.19）、5%（P=0.08）和0（P=0.97）。【结论】现有的证据提示GEMOX、GEMCAP联合化疗方案一线治疗晚期胰腺癌,有较好的应用前景,值得进一步的临床试验。     

Plasma pharmacokinetics and urinary excretion of the polyamine analogue 1, 19-bis(ethylamino)-5, 10, 15-triazanonadecane in CD2F1 mice

 The pharmacokinetics of 1, 19-bis(ethylamino)-5, 10, 15-triazanonadecane (BE-4-4-4-4) were determined in CD₂F₁ female mice after administration of i.v. bolus doses of 20 mg/kg (approximately the dose lethal to 10% of the study animals, ∼LD₁₀) as well as 15, 10, and 5 mg/kg and after s.c., i.p., or p.o. doses of 20 mg/kg. BE-4-4-4-4 in plasma and urine was derivatized with dansyl chloride and measured by gradient high-performance liquid chromatography (HPLC) with fluorescence detection. Data were modeled by noncompartmental and compartmental methods. The declines observed in plasma BE-4-4-4-4 concentrations after i.v. delivery of 20, 15, 10, and 5 mg/kg were modeled simultaneously using an interval of 2000 min between doses and were best approximated by a two-compartment, open, linear model. The time courses of plasma BE-4-4-4-4 concentrations after i.p. and s.c. delivery were fit best by a two-compartment, open, linear model with first-order absorption. Peak plasma concentrations of BE-4-4-4-4 measured following an i.v. dose of 20 mg/kg ranged between 30 and 33 μg/ml, the terminal elimination half-life was 94 min, and the volume of distribution (V_dss) was 850 ml/kg. The plasma pharmacokinetics of BE-4-4-4-4 were linear with dose. BE-4-4-4-4 (0.5 and 2.0 μM) in mouse plasma was approximately 67% protein-bound. Bioavailabilities after i.p., s.c., and p.o. delivery were 40%, 50%, and approximately 3%, respectively. Urinary excretion of parent BE-4-4-4-4 in the first 24 h after dosing accounted for less than 30% of the delivered dose. As BE-4-4-4-4 proceeds toward and undergoes clinical evaluation, the data and analytical method presented herein should prove useful in formulating a dose-escalation strategy and, possibly, evaluating toxicities encountered. Received: 25 November 1994/Accepted: 25 August 1995     

原代脐静脉内皮细胞与脐静脉内皮细胞株的生物学特性比较

目的:建立高效脐静脉内皮细胞(human umbilicalvein endothelial cell,HUVEC)体外培养方法,与HUVEC株生物学特性进行对比研究,为组织工程及相关医学基础研究提供更好的细胞来源.方法:在新鲜脐静脉内灌注2.5 g/L胰蛋白酶消化获得内皮细胞,内皮细胞培养基(endothelial cellmedium,ECM)进行培养;RPMI1640培养HUVEC株.比较两组细胞形态;免疫组化、RT-PCR检测vWF,CD144的表达;Western Blot检测两组冻存复苏后细胞vWF,CD144蛋白水平的表达.结果:原代HUVEC体外生长2～3 d可铺满单层,细胞呈梭形、饱满,漩涡状排列生长,传代后可见管腔样结构;HU-VEC株胞质多颗粒,细胞单薄.免疫组化原代HUVEC的vWF和CD144表达明显高于HUVEC株[vWF分别为(142.7±3.3),(113.6±0.7);CD144分别为(118.6±0.5),(74.2±0.4);P<0.01].在mRNA水平,原代HUVEC基因CD144(577 bp)的表达量为HUVEC株的(3.15±0.12)倍(P<0.05);vWF(434 bp)表达量为HUVEC株的(6.15±0.37)倍(P<0.05).蛋白水平冻存复苏的原代HUVEC的CD144(135ku)表达量为HUVEC株的(2.07±0.31)倍(P<0.05);vWF(210 ku)表达量为HUVEC株的(5.38±0.23)倍(P<0.05).结论:脐静脉灌注胰蛋白酶消化法配合ECM培养可迅速获取大量内皮细胞,其生物学特性明显优于HUVEC株,是组织工程及相关医学基础研究更好的细胞来源.     

Association between HLA class II locus and the susceptibility to Artemisia pollen-induced allergic rhinitis in Chinese population.

OBJECTIVE: The aim of the study was to determine whether susceptibility or resistance to Artemisia pollen-induced allergic rhinitis was associated with HLA class II DQA1, DQB1 loci.Study design and setting Forty-one subjects with allergic rhinitis and 41 healthy controls from Beijing were genotyped at HLA class II DQA1, DQB1 alleles by polymerase chain reaction amplification with sequence-specific primers-based technique. RESULTS: The allele frequencies of HLA-DQA1*0201, DQB1*0602 were lower in patients with allergic rhinitis compared with the controls (24.39% versus 46.34%, P = 0.038; 4.88% versus 26.83%, P = 0.007), and the frequency of DQA1*0302 was higher among patients than the controls (58.54% versus 14.63%, P = 0.00004, Pc = 0.0004). CONCLUSION AND SIGNIFICANCE: HLA-DQA1 and -DQB1 genes may be involved in the development of Artemisia pollen-induced allergic rhinitis. HLA-DQA1*0201, DQB1*0602 alleles may be a protective factor and DQA1*0302 may be a susceptible factor for Artemisia pollen-induced allergic rhinitis.