五氧化二砷体外对NB4细胞增殖及程序化死亡作用的研究

目的：探讨五氧化二砷（As2O3)对急性早幼粒白血病（APL）细胞NB4增殖及细胞程序化死亡的影响。方法：用细胞生长曲线，集落培养，流式细胞仪检测，DNA电泳，bcl-2蛋白和P53蛋白表达等方法观察NB4细胞增殖，凋亡和癌基因表达情况。结果：As2O5能显著地抑制NB4细胞增殖和细胞生长活力，诱导NB4细胞程序化死亡，下调p53,bcl-2蛋白表达，并能抑制造血祖细胞集落形成。结论：As2O5对APL细胞NB4具有较强的抑制和诱导凋亡作用，可能成为治疗APL有效的药物之一。     

全反式维甲酸相关多药耐药机制及其逆转的研究进展

全反式维甲酸（All-trans retinoic acid，ATRA）是急性早幼粒细胞白血病（acute promyelocytic leukemia，APL）有效的诱导分化剂，用于APL患者诱导分化治疗，完全缓解（complete recovery，CR）率可达85％-90％，其良好疗效已被国内外学者所公认。ATRA现已列为APL诱导缓解治疗的首选药物，且广泛用于其它恶性肿瘤的诱导分化治疗。     

前列腺素E_2与全反式维甲酸对人急性早幼粒细胞白血病细胞株NB_4增殖分化的协同作用

将人早幼粒细胞白血病（APL）细胞株NB4分别与前列腺素E2（PGE2）、全反式维甲酸（A－TRA）单独培养，并与一定浓度比例的上述两物质共同培养；结果发现两者在抑制细胞增殖、诱导细胞分化方面具有协同作用。联合应用可使ATRA的效应浓度比单独应用降低约20倍。这可能为对ApL，患者施行更好的口服分化治疗方案提供实验依据。     

全反式维甲酸联合丙戊酸钠诱导白血病细胞分化过程中BRD4表达变化及其作用的研究

目的探讨全反式维甲酸(ATRA)联合丙戊酸钠(VPA)诱导HL-60细胞分化过程中BRD4表达调控的分子机制。方法建立ATRA联合VPA诱导白血病细胞株HL-60的分化模型,瑞氏-吉姆萨(Wright-Gimesa)染色观察HL-60细胞形态,流式细胞术检测细胞表面分化抗原的表达,蛋白质印记从蛋白水平检测HL-60细胞经药物诱导24、48、72h后BRD4表达的变化。结果单独应用ATRA或VPA均能够明显抑制白血病细胞生长,诱导细胞分化,倒置显微镜下可观察到HL-60细胞向成熟粒细胞方向分化,细胞表面的分化抗原表达升高,联合诱导分化模型中细胞表面分化抗原表达升高更明显,两者均有统计学意义,在蛋白水平BRD4的表达逐渐降低,联合诱导分化模型中降低更明显,两者有统计学意义。结论 VPA可以明显的促进ATRA诱导白血病细胞的分化作用,BRD4基因的表达在随着分化程度的加强出现逐渐降低的趋势。     

白藜芦醇苷的合成及其维甲酸代谢阻断活性研究

目的 设计并合成白藜芦醇的多种糖苷衍生物,测定其与全反式维甲酸（ATRA）联合用药后对人类急性早幼粒细胞白血病细胞株（NB4）的诱导分化作用。方法 应用保护基策略,采用Schmidt成苷法制得目标产物;采用NBT还原法测定细胞分化百分率。结果与结论 合成了10个白藜芦醇的单糖苷化衍生物,其中,Res-3-Rha、Res-4′-Rha、Res-4′-Xyl、Res-4′-Ara是未见文献报道的新化合物;Res-3-Xyl和Res-3-Ara为首次合成的化合物,其结构均经核磁共振氢谱和质谱确证。白藜芦醇及其10个糖苷衍生物与ATRA的联合给药初筛结果表明,-Res-3-Glc、Res-3-Ara、Res-3-Rha、Res-4′-Glc、Res-4′-Ara、Res-4′-Xyl及5 μmol·L^-1的白藜芦醇和Res-4′-Rha均能明显提高ATRA的诱导细胞分化活性。     

全反式维甲酸对急性早幼粒细胞白血病出凝血及纤溶指标的影响

急性早幼粒细胞白血病（APL）是一类特殊类型的白血病，具有特征性的染色体t（15，17）易位，其特点是出血及弥散性血管内凝血（DIC）发生率高，这也是其早期死亡的主要原因。全反式维甲酸（ATRA）作为对APL诱导分化治疗的突破性药物，具有完全缓解率高、出血并发症少等优点。我们动态观察检测了70例APL患在应用ATRA后血小板计数和出血、凝血及纤溶方面指标的改变，旨在探讨其对出凝血指标的影响机制，指导临床减少APL的出血并发症，提高完全缓解率，减少其相关病死率。     

三氧化二砷治疗急性早幼粒细胞白血病过程中外周血白细胞的变化

全反式维甲酸（all—trans retinoic acid，ATRA）和三氧化二砷（As2O3）开辟了急性早幼粒细胞白血病（APL）诱导分化治疗的新篇章，大大提高了APL患的长期无复发生存率。人们早已注意到ATRA诱导治疗APL时，高白细胞血症及维甲酸综合征（RAS）有一定的发生率，同样As2O3，治疗APL时也有同样的现象。我们详细观察了As2O3治疗APL过程中外周血白细胞数量及形态的变化，并介绍临床处理及转归。     

维甲类化合物R8607对人急性早幼粒白血病NB4细胞的分化及其作用机理

目的　研究维甲类化合物R8607对人急性早幼粒白血病NB4细胞的分化诱导作用及其作用机理。方法　采用NBT还原能力和细胞形态的方法观察R8607对NB4细胞功能和形态的分化作用,采用竞争性结合的方法测定R8607和维甲酸受体的结合能力并用流式细胞术观察其对细胞周期移行的影响。结果　R8607有抑制NB4细胞的生长并能使细胞的功能和形态发生分化的作用; 对维甲酸受体的3个亚型α,β,γ均有结合作用并将NB4细胞周期移行阻断在G₁期。结论　R8607通过维甲酸受体的调节通路将NB4细胞阻断在G1期并诱导细胞发生功能和形态的分化。     

粒系集落刺激因子与急性早幼粒细胞白血病关系的研究

24例急性早幼粒细胞白血病（APL）患者，在全反式维甲酸（ATRA）治疗过程中，用ELISA法检测其血清、骨髓单个核细胞及NB4细胞用ATRA诱导前后上清液及冻融物中G－CSF水平。结果表明，在治疗前血清中G－CSF检出率为20．8％，治疗后检出率升高，在白细胞达高峰前阳性率最高，而后逐渐下降。上清液及冻融物中G－CSF均未能检出。分析表明血清G－CSF水平与外周血白细胞呈正相关（P＜0．05）。结果提示G－CSF在APL患者用ATRA治疗过程中白细胞升高的发生机制中起一定作用，APL细胞不分泌G－CSF。     

维A酸联合亚砷酸治疗急性早幼粒细胞白血病

急性早幼粒细胞白血病（APL）是以早幼粒细胞增生为主的急性白血病,为FAB分型的M3型,起病急骤。维A酸（ATRA）诱导化疗虽能使90％患者达到完全缓解,但由于体内残留少量白血病细胞增殖,仍会有60％最终复发…,一直是临床上致力于解决的难题。近年来,笔者对APL患者应用三联疗法（ATRA＋亚砷酸＋化疗药物）,取得良好效果,现报告如下。     

全反式维甲酸对小细胞肺癌细胞的分化诱导作用研究

目的探讨全反式维甲酸(all-trans-retinoic acid,ATRA)对小细胞肺癌细胞的分化诱导作用。方法选择NCI-H446细胞株MTT法检测细胞体外增殖能力,电镜下观察形态,流式细胞仪进行细胞周期分析。RT-PCR法分析维甲酸受体亚型基因。结果全反式维甲酸诱导小细胞肺癌NCI-H446细胞株,细胞增殖能力下降,凋亡增加,更多的细胞被阻止于G1/G0期。结论全反式维甲酸诱导分化能有效抑制小细胞肺癌NCI-H446细胞株的增殖,促进凋亡。     

全反式维甲酸和阿糖胞苷对HL-60细胞端粒酶活性的影响

目的 探讨全反式维甲酸（all trans-retinoic acid,ATRA)和阿糖胞苷（Ara-c)诱导人早幼粒细胞白血病细胞凋亡和分化的机制。方法 采用PCR－ELISA方法观察ATRA和Ara-c对HL－60细胞端粒酶活性的影响。结果 ATRA和Ara-c在诱导HL－60细胞凋亡和分化过程中均可使端粒酶 活性减低。结论 ATRA和Ara-c抑制HK－60细胞端粒酶的活性,可能是其诱导白血病细胞凋亡和分化的重要机制之一。     

All-trans retinoic acid induced differentiation of rat mammary epithelial cells cultured in serum-free medium

Retinoids are applied to not only cancer prevention but also cancer chemotherapy by stimulating differentiation of cells. We studied differentiation inducing effect of all-trans retinoic acid (ATRA) by studying proportion of high dense fractions of stem-like cells and the size of S phase fraction in cell cycle. From mammary organoids obtained from 7- to 8-week old F344 female rat mammary gland, we cultured rat mammary epithelial cells (RMEC) and treated physiological doses of 10⁻⁶, 10⁻⁷, and 10⁻⁸ M ATRA from the first day and then cultured for 4, 7, and 14 days. After that, immunostaining was performed using peanut agglutinin (PNA) and anti-Thy-1.1 monoclonal antibody (Thy-1.1) that can be used as markers of differentiation. We separated four different cell subpopulations by flow cytometry: cells negative to both reagents (B-), PNA-positive cells (PNA+), Thy-1.1-positive cells (Thy-1.1+), and cells positive to both reagents (B+). We observed continuous decreases of high dense fractions of stem-like cells (PNA+ subpopulations) for 14 days and as much decreases as high doses of ATRA, which were thought to be proportional to doses of ATRA. We labeled RMEC with bromodeoxyuridine and investigated cell cycle fractions that went through S phase. We observed a tendency of decrease of S phase fraction with time in culture, which is though to be related to continuous decreases of PNA+ subpopulations and inhibitory role of ATRA on cell cycle. These results suggest that physiological doses of ATRA could stimulate differentiation of RMEC and convert stem-like RMEC to differentiated cells in SFM for a relatively long period of 14 days.     

Pharmacological evaluation of various metabolites and analogues of valproic acid. Anticonvulsant and toxic potencies in mice

Thirty-two metabolites and analogues of the antiepileptic drug valproic acid (2-propylpentanoic acid; VPA) were tested for anticonvulsant and toxic effects in mice, in an attempt to find out if any of these compounds were superior to valproic acid. Valproic acid and ethosuximide, another clinically established antiepileptic drug, were included in these studies for comparison. After intraperitoneal administration, the anticonvulsant potency of the various drugs was determined in three seizure tests: the threshold for maximal electroconvulsions, the maximal electroshock seizure test and seizures induced by subcutaneous injection of pentylenetetrazol. For the most potent compounds, median minimal neurotoxic doses (TD50S) and LD50S (after i.p. and i.v. injection) were determined. Valpramide, the primary amide of valproic acid, proved to be the most potent compound in the three seizure tests, used, being 2-5 times as potent as valproic acid, but valpramide was also considerably more sedative and toxic than valproic acid or ethosuximide. Of the metabolites of valproic acid tested, the unsaturated compounds 4-en-valproic acid (4-en-VPA) and the trans-isomer of 2-en-valproic acid (2-en-VPA) were most potent and, depending on the seizure test used, reached 60-100% of the efficacy of the parent drug. Both metabolites had LD50 values which were similar or greater than those of valproic acid but they were more sedative than the parent compound.(ABSTRACT TRUNCATED AT 250 WORDS)     

Novel mutual prodrug of retinoic and butyric acids with enhanced anticancer activity

Acyloxylalkyl esters of retinoic acid and small carboxylic acids (C3-5) were evaluated for anticancer activity. The derivative of butyric acid (BA) and all-trans-retinoic acid (ATRA)-retinoyloxymethyl butyrate (RN1)-acting as a mutual prodrug was a more potent inducer of cancer cell differentiation and inhibitor of proliferation than the parent acids. ED50 of RN1 for differentiation induction in HL-60 was over 40-fold lower than that of ATRA. The differentiating activity of ATRA compared to that of the acyloxylalkyl esters derived from butyric (RN1), propionic (RN2), isobutyric (RN3), and pivalic (RN4) acids was found to be: RN1 > RN2 > RN3 > ATRA approximately RN4. This observation implies that the activity of the prodrugs depends on the specific acyl fragment attached to the retinoyl moiety, and the butyroyl fragment conferred the highest potency. The IC50 values for inhibition of Lewis lung (3LLD122) and pancreatic (PaCa2) carcinoma cell line colony formation elicited by RN1 were significantly higher than those of ATRA. In addition to its superiority over ATRA or BA as growth inhibitors of the above cell lines, RN1 was also able to overcome the resistance to ATRA in 3LLD122 cells.     

Design,synthesis and pharmacological evaluation of caffeic acid phenethyl ester acylation as multifunctional neuroprotective agents against oxidative stress injury

4-Acylated or 3,4-diacylated caffeic acid phenethyl ester (CAPE) was prepared as prodrug to improve its stability and lipid solubility. Their neuroprotective activities were assessed by H2O2 model and 6-OHDA model. The results showed that target compounds displayed positive abilities to protect PC12 nerve cells from oxidative stress injury, superior to that of CAPE. Additionally, target compounds showed high blood-brain barrier permeability.     

丙戊酸钠及其代谢产物与肝损伤的相关性分析

目的研究丙戊酸钠及3个代谢产物(2-丙基-4-五烯酸、3-羟基丙戊酸、5-羟基丙戊酸)对肝损伤参考指标的相关性分析。方法共收集328例癫痫患者血样,其中,123例肝功能异常癫痫患者血样为试验组,205例肝功能正常癫痫患者血样为对照组,采用LC-MS/MS方法测定两组血样(丙戊酸钠及代谢产物)的血药浓度,通过ROC曲线分析丙戊酸及其代谢产物浓度对肝功能异常的诊断价值。结果肝功能异常组患者丙戊酸钠及其3个代谢产物平均血药浓度均高于对照组,差异有统计学意义(P<0.05)。丙戊酸钠及其代谢产物的血药浓度均可作为诊断肝损伤的参考指标,5-羟基丙戊酸比丙戊酸钠有更好的诊断价值。结论丙戊酸钠代谢产物与肝毒性有关,能够作为肝损伤的诊断指标,可将其应用于临床,为丙戊酸钠有效给药提供参考。