农药Atonik的致癌性

目的　探讨新植物生长调节剂Ａｔｏｎｉｋ致癌的潜在可能性 ,为其安全使用 ,农药登记提供技术依据。方法　大鼠ＳＤ种 ,6～ 7周龄 ,每组 10 0只 ,雌雄各半。 3个染毒组和对照组分别每天饮用含Ａｔｏｎｉｋ 6 0 0、15 0、38和 0ｍｇ Ｌ的饮水 ,连续 2年。结果　高、中、低染毒组和对照组的 2年存活率分别为6 7%、6 2 %、6 6 %和 6 6 %。 2年内共检出肿瘤 93例 ,肿瘤检出率分别为 2 2 2 %、2 2 0 %、2 2 4%和 2 9 3 % ,瘤样增生检出率为 7 9%、5 2 %、5 8%和 7 9% ,均无明显的组间差异。 93例肿瘤中 ,良性 78例 ,恶性 15例 ,其中乳腺瘤 3…     

甲状腺肿瘤细胞中P53、P63蛋白表达的比较研究

目的探讨P5 3和P6 3蛋白在甲状腺肿瘤各型细胞中的表达情况 ,并观察它们的相似及差异性。方法采用免疫组织化学 (EnvisionTM)法检测 6 0例甲状腺肿瘤中P5 3和P6 3蛋白的表达。结果P5 3蛋白在甲状腺腺瘤及腺癌中的阳性表达率分别为 0 0 0 % (0 /30 )和 2 0 0 % % (6 /30 ) ,两者比较差异有显著意义 (P <0 0 5 ) ;P6 3蛋白在甲状腺腺瘤及腺癌中的阳性表达率分别为 4 6 7% (14 /30 )和 2 0 0 % (6 /30 ) ,两者比较差异有显著意义 (P <0 0 5 ) ;P5 3在甲状腺乳头状腺瘤及滤泡状腺瘤中均不表达 ,而在甲状腺乳头状腺癌及滤泡状腺癌中的阳性表达率分别为 13.3% (2 /15 )和 33.3% (5 /15 ) ;P6 3蛋白在甲状腺乳头状腺瘤及滤泡状腺瘤中的阳性表达率分别为 6 0 .0 % (9/15 )和 33.3% (5 /15 ) ,而在甲状腺乳头状腺癌及滤泡状腺癌中的阳性表达率均为 2 0 .0 % (3/15 )。结论P5 3和P6 3蛋白在甲状腺肿瘤细胞中表达的差异 ,可能提示P5 3和P6 3蛋白在肿瘤抑制和组织发育中起着不同的作用。     

注射用盐酸地尔硫(艹卓)的HPLC测定

采用 HPL C法测定注射用盐酸地尔硫的含量及有关物质。色谱柱为 Diamonsil C1 8( 15 0 mm× 4.6 m m,5μm ) ,0 .1m ol/ L醋酸钠缓冲液 ( p H6 .2 ) -甲醇 -乙腈 ( 5∶ 6∶ 6 )为流动相 ,检测波长为 2 40 nm。检测限为 0 .2 ng,线性范围为 6～ 14μg/ ml( r=0 .9998) ,RSD为 0 .11% ,平均回收率为 10 0 .2 % ,RSD为 0 .5 2 %。     

注射用雷替曲塞的HPLC法测定

用HPLC法测定注射用雷替曲塞的含量及有关物质。采用PhenomenexC18柱 ,流动相为 0 15mol/L乙酸水溶液-甲醇 (5 0∶5 0 ) ,检测波长为 2 2 8nm ,流速为 0 8ml/min。雷替曲塞在 0 0 2 97～ 0 0 6 93mg/ml之间呈良好的线性关系 (r =0 9998) ,平均回收率为 99 4 % ,(RSD =0 6 6 % )     

精神分裂症300例复发因素调查分析

精神分裂症易复发 ,为弄清其原因 ,我们对 30 0例复发的精神分裂症进行调查分析 ,现报告如下。1　资料与方法1.1　临床资料30 0例患者中男 171例 ,占 5 7% ,女 12 9例 ,占 43% ;年龄 18～ 6 4岁 ,平均年龄 (34± 10 )岁。已婚 15 0例 ,占 5 0 % ;未婚 90例 ,占 30 % ;离婚 6 0例 ,占 2 0 %。农民 12 0例 ,占40 % ;工人 12 0例 ,占 40 % ;其他职业 6 0例 ,占 2 0 %。有阳性家族史者 45例 ,占 15 %。病程 0 .5～ 30 a,平均 (15± 8)a。已住院次数 1～ 2次者 180例 ,占 6 0 % ;3次以上者 12 0例 ,占 40 %。1.1.1　定期复查的时间与复发 :定期复…     

248株念珠菌和新型隐球菌对四种抗真菌药的敏感性

目的　检测 2 48株念珠菌和新型隐球菌对四种抗真菌药氯康唑、两性霉素B、5 氟胞嘧啶和酮康唑的耐药状况。方法　收集临床菌株新型隐球菌 6 0株和念珠菌 188株 ,采用NCCLSM2 7 A微量稀释法测定新型隐球菌和念珠菌临床分离株对四种抗真菌药物的最低抑菌浓度 (MIC)。结果　对 16 0株白念珠菌 ,氟康唑的MIC为 0 .0 6～≥ 6 4μg/ml,敏感 135株 ( 84.38% ) ,中度敏感 10株 ( 6 .2 5 % ) ,耐药 15株 ( 9.37% ) ;5 氟胞嘧啶的MIC为 0 .0 6～ 16 μg/ml ,敏感 15 6株 ( 97.5 % ) ,中度敏感 2株 ( 1.2 5 % ) ,耐药 2株 ( 1.2 5 % ) ;两性霉素B的MIC为 0 .0 6～ 4μg/ml全部敏感 ;酮康唑的MIC为 0 .0 3～ 32 μg/ml ,敏感 93.75 % ,中度敏感 6 .2 5 %。对 6 0株新型隐球菌 ,氟康唑的MIC 1～≥ 6 4μg/ml,敏感 35株 ( 5 8.4% ) ,中度敏感 2 0株 ( 33.3% ) ,耐药 5株( 8.3% ) ;5 氟胞嘧啶MIC 0 .2 5～ 8μg/ml,敏感 5 3株 ( 88.3% ) ,中度敏感 7株 ( 11.7% ) ;两性霉素B的MIC为 0 .0 6～ 0 .5 μg/ml全部敏感 ;酮康唑MIC 0 .0 3～ 4μg/ml全部敏感。热带念珠菌和光滑念珠菌对氟康唑耐药率分别为 10 %和2 2 .2 %。结论　新型隐球菌和念珠菌对氯康唑的耐药率趋高 ,临床不容忽视。     

HPLC法测定市售连翘中连翘酯苷的含量

建立了连翘中连翘酯苷的HPLC测定方法 ,采用TIANHEC18柱 (15 0mm× 4 6mm) ,流动相为乙腈—水—醋酸 (15∶85∶0 2 ) ,检测波长为 332nm ,流速为 1 0ml·min-1。连翘酯苷在 0 2 0 2～ 16 2 μg范围内线性关系良好 (r =0 9999) ,平均回收率为 10 0 5 0 % ,RSD =1 6 1%。并测定了陕西市场上连翘药材中连翘酯苷的含量 ,对其质量进行评价。     

林可霉素利多卡因凝胶含量测定方法的改进

目的　改进林可霉素利多卡因凝胶的含量测定方法。方法　采用高效液相色谱法。色谱柱为AlltimaC18柱 (15 0mm× 4 6mm ,5 μm) ,流动相为 0 0 5mol/L硼砂溶液 (用 85 %磷酸调pH值至 5 0 ) 甲醇 乙腈 (6 0∶37∶3) ;检测波长为 2 14nm ;流速为 1 0ml/min。结果　盐酸林可霉素和盐酸利多卡因分别在浓度为 0 0 2～ 0 4 0mg/ml(r=0 9999,n =6 )和 0 0 1～ 0 32mg/ml(r =1 0 0 0 0 ,n =6 )范围内呈良好线性关系。盐酸林可霉素和盐酸利多卡因的平均回收率分别为 10 0 3% (RSD为 0 6 2 % ,n=6 )、10 1 1% (RSD为 0 4 6 % ,n =6 )。结论　该方法操作简便 ,结果准确 ,重现性好。     

HPLC测定甘芪胃疡宁颗粒中芍药苷的含量

目的　建立甘芪胃疡宁颗料中芍药苷含量测定方法。方法　用HPLC测定 ,分析柱为ODS柱 (2 0 0mm×4 6mm ,5 μm) ,流动相为 0 0 5 %磷酸二氢钾溶液 -乙晴 (85∶15 ) ,检测波长 2 30nm ,流速为 0 9ml·min-1。结果　线性范围 0 2 31～ 1 15 8μg(r=0 9999)。平均回收率 98 2 7% ,RSD =0 86 % (n =6 )。 结论　该法简便、灵敏、准确。可用于甘芪胃疡宁颗粒的质量控制     

恶性肿瘤患者HBV、HCV、HAV血清标志物检测分析

采用ELISA方法检测了恶性肿瘤患者 ,2 15 3例 ,正常人 5 79例的血清HBV、HCV、HAV标志物 (乙肝系列五项 ,抗 -HCV、抗 -HAVIgM)。经统计学处理 ,采用X2 检验 ,结果显示 :恶性肿瘤患者单项抗 -HBs阳性率 2 8 3% (60 9/ 2 15 3) ,低于正常人4 0 9% (2 37/ 5 79) ,差别显著 (P <0 0 5 )。HBV抗原阳性率 11 8% (2 5 3/ 2 15 3) ,高于正常人 8 6% (5 0 / 5 79) ,差别显著 (P <0 0 5 )。抗 -HCV ,肿瘤患者阳性率为2 6% (5 7/ 2 15 3) ,高于正常人 0 5 (3/ 5 79) ,差别非常显著 (P <0 0 1)。有HCV与HBV重叠感染 ,抗 -HAVIgM检出率极低 ,亦有HAV亦与HBV重叠感染 ,但未发现HBV、HCV、HAV三者同时感染的病例     

HPLC法同时测定腹泻康片中秦皮甲素和秦皮乙素的含量

目的建立同时测定腹泻康片中秦皮甲素和秦皮乙素含量的方法。方法采用高效液相色谱法,色谱柱:Kromasil C18柱(4.6 mm×250 mm,5μm);流动相:乙腈-0.1%磷酸(8∶92);检测波长:334 nm;流速:1 m L·min-1;柱温:30℃。结果秦皮甲素线性范围为47.2⁴72 ng,r=0.999 0,秦皮乙素线性范围为45.6472 ng,r=0.999 0,秦皮乙素线性范围为45.6⁴56 ng,r=0.999 2,秦皮甲素和秦皮乙素平均加样回收率分别为101.09%和100.51%。结论本方法简便、准确、可靠,适用于腹泻康片中秦皮甲素和秦皮乙素的质量控制。     

HPLC法同时测定消炎片中绿原酸、秦皮乙素和咖啡酸的含量

目的:建立HPLC法同时测定消炎片中绿原酸、秦皮乙素和咖啡酸的含量.方法:采用Agilent Eclipse XDB-C18 (250mm×4.6 mm,5μm)色谱柱,以甲醇-乙腈-0.04％磷酸溶液为流动相,梯度洗脱;检测波长为328 nm;流速1.0ml·min -1;进样量20μl;柱温35℃.结果:绿原酸、秦皮乙素和咖啡酸的线性范围分别为1.616 ～40.400μg·ml-1(r=0.999 9,n=7)、5.164～129.100μg ·ml-1(r=1.000 0,n=7)和1.344～ 26.880 μg·ml-1(r=1.000 0,n=7);提取回收率分别为99.08％(RSD=0.6％,n=6)、98.81％(RSD =0.4％,n=6)和98.92％(RSD=0.9％,n=6).结论:本法操作简便、快速、结果准确.     

Induction of apoptosis by esculetin in human leukemia cells

Esculetin, a coumarin compound, has been shown to exhibit antioxidant and anti-inflammatory effects. In the present study, esculetin was found to inhibit the survival of human promyelocytic leukemia HL-60 cells in a concentration-dependent and time-dependent manner. HL-60 cells underwent internucleosomal DNA fragmentation and morphological changes characteristic of apoptosis after a 24-h treatment with esculetin (100 microM). Flow cytometric analysis showed that the hypodiploid nuclei of HL-60 cells were increased to 40.93% after a 36-h treatment with esculetin (100 microM). Further investigation showed that esculetin induced the release of cytochrome c from mitochondria into cytosol in a time-dependent and concentration-dependent manner. Moreover, esculetin application reduced Bcl-2 protein expression to 58% after 9 h as compared with that time at 0. Cysteine protease 32 kDa proenzyme (CPP32), a caspase 3, was activated and its substrate, poly (adenosine diphosphate-ribose) polymerase, was cleaved after a 24-h treatment of HL-60 cells with esculetin. These data suggest that esculetin induces apoptosis in human leukemia cells by increasing cytosolic translocation of cytochrome c and activation of CPP32.     

Inhibitory effects of esculetin on melanin biosynthesis

To investigate the structure-activity relationship of coumarins for the inhibitory activity on mushroom tyrosinase, the 50% inhibitory concentration (IC50 values) of 18 coumarins and four cinnamic acid derivatives were measured. Among these compounds, esculetin had the strongest inhibitory activity (IC50=43 microM) on mushroom tyrosinase. Introduction of a hydroxy group to the C6 and C7 positions of the coumarin ring and no substitution on the lactone ring played an important role in the expression of the strong inhibitory activity of esculetin. We performed further studies to estimate the in vitro inhibitory effects of esculetin on melanogenesis. Esculetin 5 microM significantly suppressed melanin production in murine B16 melanoma cells without affecting cell growth. Furthermore, the number of 3,4-dihydroxyphenylalanine (DOPA)-positive melanocytes in the split-epidermal sheets treated with 0.05% or 0.1% esculetin was significantly lower than that in the control. From these results, it is suggested that esculetin has inhibitory effects on tyrosinase activity in vitro. However, further detailed studies are necessary to understand the inhibitory mechanism of esculetin.     

Anti-oxidative and photo-protective effects of coumarins isolated fromFraxinus chinensis

Free radicals and reactive oxygen species (ROS), which are generated by UV irradiation, may cause serious injury to skin cell membranes, DNA and functional proteins. In addition, these agents stimulate the expressions of matrix metalloproteinases (MMPs), which can degrade most components of the extracellular matrix (ECM), including collagen. In order to develop new anti-photoaging agents, five major components from the extract of Fraxinus chinensis extract (FCE) were identified. Two of the major components of FCE were found to be esculin (11.2%) and esculetin (1.9%). FCE (IC50: 50.0 microg/mL 1, 1-diphenyl-2-picrylhydrazyl (DPPH); 19.8 microg/mL, superoxide anion radical) and esculetin (IC50: 2.1 microg/mL DPPH; 0.6 microg/mL, superoxide anion radical) showed strong antioxidative activities. Of the compounds tested, esculetin showed the strongest scavenging activity against DPPH radicals, followed by superoxide anions from the xanthine/xanthine oxidase system. The intracellular ROS scavenging activity showed that oxidation of 5-(6-)-chloromethyl-2', 7'-dichlorodihydrofluorescein diacetate (CM-H2DCFDA) was effectively inhibited by esculetin, with potent free radical scavenging activity was also shown in UVB-irradiated human dermal fibroblasts (HDFs). Moreover, treatment of UVA-irradiated HDFs with esculetin resulted in dose-dependent decreases in the expression levels of MMP-1 mRNA and protein. From these results, FCE and one of its components, esculetin, were predicted to be potentially useful as ingredients in cosmetics for protecting against photoaging.     

Differential effects of esculetin and daphnetin on in vitro cell proliferation and in vivo estrogenicity

Esculetin (6,7-dihydroxycoumarin) and daphnetin (7,8-dihydroxycoumarin) are secondary metabolites of plants used in folk medicine. These compounds have showed great antiproliferative activity in several tumor cell lines and have been proposed as potential anticancer agents. However, the estrogenic potential of these two compounds has to date not been reported. The present study compared esculetin and daphnetin on the inhibition of cell proliferation and cell cycle progression of the MCF-7 estrogen-responsive human carcinoma cell line. In vivo and in vitro estrogenic activity for both compounds was also evaluated. Esculetin inhibited cell proliferation after 72 h exposure (IC50=193 ± 6.6 μM), while daphnetin evidenced inhibiting effects starting at 24-h exposure (72 h, IC50=73 ± 4.1 μM). Both effects showed changes in cyclin D1 gene expression. In non-estrogenic conditions (E-screening assay), esculetin produced biphasic response on proliferation of the MCF-7 cells; at 10(-8)-10(-6)M, concentrations induced proliferative effects as EC50=4.07 × 10(-9)M (E(2)=2.91 × 10(-12)M); at higher concentrations (10(-5)-10(-4)M), cell proliferation was inhibited. Relative proliferative effect at E(2) was 52% (E(2)=100), relative proliferative potency was 0.072 (E(2)=100). Additionally, esculetin tested in vivo showed estrogenic effects at 50-100mg/kg doses; relative uterotrophic effect at E(2) was 37%, with relative uterotrophic potency registered at 0.003. In contrast, daphnetin did not induce estrogenic effects in vitro or with in vivo models. The low estrogenic activity of esculetin could prove useful in postmenopausal therapy but not as a safe antitumor agent in estrogen-dependent tumors. Daphnetin-based antiproliferative selectivity with MCF-7 cells showed that daphnetin is a promising antitumoral agent also acting on estrogen dependent tumors.     

The effect of interferon-α on the expression of cytochrome P450 3A4 in human hepatoma cells

Esculetin (6,7-dihydroxy coumarin), is a potent antioxidant that is present in several plant species. The aim of this study was to investigate the mechanism of protection of esculetin in human hepatoma HepG2 cells against reactive oxygen species (ROS) induced by hydrogen peroxide. Cell viability, cell integrity, intracellular glutathione levels, generation of reactive oxygen species and expression of antioxidant enzymes were used as markers to measure cellular oxidative stress and response to ROS. The protective effect of esculetin was compared to a well-characterized chemoprotective compound quercetin. Pre-treatment of HepG2 cells with sub-lethal (10-25 μM) esculetin for 8 h prevented cell death and maintained cell integrity following exposure to 0.9 mM hydrogen peroxide. An increase in the generation of ROS following hydrogen peroxide treatment was significantly attenuated by 8 h pre-treatment with esculetin. In addition, esculetin ameliorated the decrease in intracellular glutathione caused by hydrogen peroxide exposure. Moreover, treatment with 25 μM esculetin for 8 h increased the expression of NAD(P)H: quinone oxidoreductase (NQO1) at both protein and mRNA levels significantly, by 12-fold and 15-fold, respectively. Esculetin treatment also increased nuclear accumulation of Nrf2 by 8-fold indicating that increased NQO1 expression is Nrf2-mediated. These results indicate that esculetin protects human hepatoma HepG2 cells from hydrogen peroxide induced oxidative injury and that this protection is provided through the induction of protective enzymes as part of an adaptive response mediated by Nrf2 nuclear accumulation.     

香附中mesocyperusphenol A的含量测定研究

目的：建立香附中 mesocyperusphenol A 的超高效液相色谱（UHPLC）含量测定方法,为香附的合理质量控制提供依据。方法采用 Poroshell 120 EC - C18色谱柱（3.0 mm ×150 mm,1.8μm）,流动相乙腈-水（28∶72,V∶V）,检测波长：325 nm。结果 Mesocyperusphenol A 在0.784~7.84μg·mL -1范围内线性关系良好,r ＝0.9997（n＝6）,加样回收率为101.6%,RSD ＝1.39%（ n ＝9）;6批香附药材中 mesocyperusphenol A 的含量为3.72~10.00μg·g -1。结论 Mesocyperusphenol A 为香附子药材中的特征活性成分,所建立的测定方法操作简便可靠,可为香附药材的质量控制提供依据。     

HPLC法测定复方当归注射液中两种成分的含量

目的：建立同时测定复方当归注射液中羟基红花黄色素 A 和阿魏酸含量的方法。方法采用高效液相色谱法。色谱柱为 Inertsil ODS -3（4.6 mm ×250 mm,5μm）,流动相为甲醇-0.1％磷酸溶液（35∶65,V/ V）,流速为1.0 mL·min -1,检测波长为403 nm（0～10 min,羟基红花黄色素 A）、321 nm（10～20 min,阿魏酸）,柱温为35℃,进样量为10μL。结果羟基红花黄色素 A、阿魏酸的进样量分别在0.0426～1.7060、0.0112～0.4480μg（r＝1.000）范围内与相应峰面积呈良好的线性关系;二者精密度、稳定性、重复性试验的 RSD 均＜2.0％;平均加样回收率分别为99.76％、100.50％,RSD 分别为1.35％、1.19％（n ＝9）。结论本法简便、快速、重复性好,可用于复方当归注射液的质量控制。     

RP-HPLC法测定清胰利胆颗粒中绿原酸和芍药苷的含量

目的建立同时测定清胰利胆颗粒中绿原酸和芍药苷含量的方法。方法采用HPLC法。色谱柱:Kromasil C18柱(250 mm×4.6 mm,5μm),流动相:乙腈(A)-体积分数为0.1%的磷酸水溶液(B),梯度洗脱程序:0 min-10%(φ)A、9 min-13%(φ)A、13 min-13%(φ)A、14 min-16%(φ)A、22min-17%(φ)A、30 min-17%(φ)A、40 min,40%(φ)A,流速:1.0 mL.min-1,检测波长:327 nm(0～14 min)和230 nm(14～40 min)。结果绿原酸和芍药苷质量浓度分别在5.2～51.6 mg.L-1(r=0.999 6,n=6)、9.9～99.2 mg.L-1(r=0.999 8,n=6)内与峰面积线性关系良好,平均回收率分别为100.5%(RSD=1.9%,n=6)、100.2%(RSD=2.2%,n=6)。结论该方法可作为清胰利胆颗粒质量控制方法之一。