HPLC法测定泌炎康胶囊中绿原酸的含量

目的：建立高效液相色谱法（HPLC）测定泌炎康胶囊中绿原酸含量的方法。方法：采用YMC-C18柱。流动相为乙腈-0.4%磷酸（13：87）；检测波长为324nm。结果：该方法的线性范围为0.08-0.42μg（r=0.9999,n=6)平均回率为100.35%，RSD=1.37%。结论：本方法简便，准确，灵敏度高，重现性好，可用于该制剂成分的含量测定。     

金粉蕨素拮抗氧化损伤所抑制的内皮细胞增殖的作用及其机制

目的　探讨金粉蕨素拮抗Menadione氧化损伤所抑制的内皮细胞增殖的作用及其机制。方法　以Menadione(O- 2 )损伤人脐静脉内皮细胞作为氧化损伤模型 ,采用MTT法和细胞计数法 ,观察不同浓度金粉蕨素对Menadione损伤内皮细胞生长抑制率的影响 ;利用硝酸还原酶法测定培养液中NO含量 ;以Westernblot检测细胞eNOS活性及磷酸化ERK1 / 2的表达。结果　金粉蕨素保护组与损伤组相比 ,内皮细胞的生长抑制率明显降低 ,培养液中NO含量增高 ,eNOS活性增强 ,磷酸化ERK1 / 2表达上调。结论　NO和ERK1 / 2通路可能介导了金粉蕨素拮抗Menadione氧化损伤所抑制内皮细胞增殖的保护作用     

上海骨关节疾病患者口服吲哚美辛不良反应危险因子筛选

目的　在上海骨关节疾病患者中筛选吲哚美辛引起不良反应的危险因素。方法　利用回顾性流行病学调查方法 ,调查服用吲哚美辛的骨关节疾病患者的一般情况、原发疾病、疾病家族史、服用非甾体抗炎药前生活质量评估、饮食习惯、生活方式、非甾体抗炎药服用情况和不良反应发生情况等 ,通过单因素和多因素分析从中筛选吲哚美辛不良反应发生的相关因素。结果　上海骨关节疾病患者中口服吲哚美辛的不良反应发生率为 48 1%。因子“合并药物治疗”(比值比 0 166,95%可信区间 0 0 3 7～ 0 74)、“饮酒史”(比值比 0 795,95%可信区间 0 675～ 0 93 7)、“来自于健康问题的何种程度的紧张或压力会影响你的生活” (比值比0 917,95%可信区间 0 848～ 0 992 )评分的降低分别使口服吲哚美辛不良反应发生的危险增高。而因子“与 6mon前比较 ,服用非甾体抗炎药前的健康状况如何” (比值比1 2 3 6,95%可信区间 1 0 2 2～ 1 496)评分的升高将使口服吲哚美辛不良反应发生的危险增高。结论　“合并药物治疗”、“饮酒史”、“与 6mon前比较 ,服用非甾体抗炎药前的健康状况如何”是吲哚美辛不良反应发生的危险因子     

氟他胺与2-羟基氟他胺对大鼠前列腺增生的抑制作用(英文)

目的 :研究氟他胺与 2 羟基氟他胺对诱导的前列腺增生大鼠前列腺萎缩和细胞凋亡的作用。方法 :SD大鼠去势 ,皮下注射丙酸睾丸素诱导前列腺增生。前列腺增生大鼠随机分成以下 5组 :对照组、氟他胺组 5 0与 1 0 0mg·kg-1,2 羟基氟他胺组2 5与 5 0mg·kg-1,用药时间为 1 0d。比较前列腺各侧叶湿重 ,以HE染色及TUNEL染色法观察细胞凋亡并计算凋亡发生率。结果 :氟他胺及 2 羟基氟他胺可明显降低前列腺增生大鼠前列腺各侧叶湿重 ,差异有显著意义 ,并可观察到明显的凋亡细胞。结论 :氟他胺与 2 羟基氟他胺可诱导前列腺增生大鼠的前列腺萎缩和细胞凋亡     

13例肝局灶性结节增生的CT和MR诊断

目的 探讨肝脏局灶性结节增生(FNH)的CT和MR表现及诊断价值。方法 回顾分析13例经手术病理证实为肝脏FNH的CT和MR表现，研究其影像的诊断价值。结果 13例FNH的CT和MR中7例表现较为典型，CT平扫等密度，T1WI为等信号，T2WI为等信号或稍高信号；部分病灶可见中央区低密度瘢痕，增强动脉期除中央瘢痕外均明显强化，门静脉期常见明显强化，延时期多为等密度，瘢痕可稍有强化；术前作出正确诊断。6例FNH表现不典型，CT低密度灶，T1WI为低信号，T2WI为高信号，无瘢痕和轻度强化；术前3例诊断为恶性肿瘤，3例诊断为血管瘤。结论 FNH的CT和MR表现在术前进行综合分析，多数可明确诊断。     

Validation of serotonin (5-hydroxtryptamine) as an in vitro substrate probe for human UDP-glucuronosyltransferase (UGT) 1A6.

Investigation of human UDP-glucuronosyltransferase (UGT) isoforms has been limited by a lack of specific substrate probes. In this study serotonin was evaluated for use as a probe substrate for human UGT1A6 using recombinant human UGTs and tissue microsomes. Of the 10 commercially available recombinant UGT isoforms, only UGT1A6 catalyzed serotonin glucuronidation. Serotonin-UGT activity at 40 mM serotonin concentration varied more than 40-fold among human livers (n = 54), ranging from 0.77 to 32.9 nmol/min/mg of protein with a median activity of 7.1 nmol/min/mg of protein. Serotonin-UGT activity kinetics of representative human liver microsomes (n = 7) and pooled human kidney, intestinal and lung microsomes and recombinant human UGT1A6 typically followed one enzyme Michaelis-Menten kinetics. Serotonin glucuronidation activity in these human liver microsomes had widely varying V(max) values ranging from 0.62 to 51.3 nmol/min/mg of protein but very similar apparent K(m) values ranging from 5.2 to 8.8 mM. Pooled human kidney, intestine, and lung microsomes had V(max) values (mean +/- standard error of the estimates) of 8.8 +/- 0.4, 0.22 +/- 0.00, and 0.03 +/- 0.00 nmol/min/mg of protein (respectively) and apparent K(m) values of 6.5 +/- 0.9, 12.4 +/- 2.0, and 4.9 +/- 3.3 mM (respectively). In comparison, recombinant UGT1A6 had a V(max) of 4.5 +/- 0.1 nmol/min/mg of protein and an apparent K(m) of 5.0 +/- 0.4 mM. A highly significant correlation was found between immunoreactive UGT1A6 protein content and serotonin-UGT activity measured at 4 mM serotonin concentration in human liver microsomes (R(s) = 0.769; P < 0.001) (n = 52). In conclusion, these results indicate that serotonin is a highly selective in vitro probe substrate for human UGT1A6.     

中国汉族人群遗传性聋基因座位STR位点的遗传多态性研究

目的 研究中国汉族人群10个遗传性聋基因座位的短串连重复序列(STR)的遗传多态性。方法 应用PCR方法对10个位点进行扩增，变性聚丙烯酰胺凝胶电泳分离，记录基因型。采用PPAP软件计算正常人各STR位点等位片段频率、基因型频率、预期基因型频率、多态信息量(PIC)和Hardy-Weinberg平衡吻合性检验。结果 中国汉族人群D6S287、D8S1132、D13S1275、D15S130、D1S2726、D4S3038、D2S2380、D22S282、D14S1042、D7S529各位点分别检出10个、9个、8个、7个、7个、7个、6个、6个、6个及5个等位片段，多态性分布均符合Hardy-Weinberg平衡定律。各位点PIC值分别为0．879、0．854、0．864、0．821、0．686、0．794、0．764、0．732、0．773、0．712；杂合度均高于0．7。结论 中国汉族人群10个位点STR均具有较高的杂合度和多态信息量，是较理想的遗传标记。     

Interleukin-12 enhances the sensitivity of human osteosarcoma cells to 4-hydroperoxycyclophosphamide by a mechanism involving the Fas/Fas-ligand pathway.

Cyclophosphamide (CY) and its derivative ifosfamide are alkylating agents used to treat osteosarcoma (OS). The purpose of these studies was to determine whether alkylating agents affect the expression of Fas ligand (FasL) and whether interleukin 12 enhances the sensitivity of human OS cells to alkylating agents. 4-Hydroperoxycyclophosphamide (4-HC), the preactivated CY compound, and 4-hydroperoxydidechlorocloclophosphamide (4-HDC), its nonalkylating analogue, human OS LM6 cells, and a clone of cells derived by transfection with the interleukin 12 gene (LM6-#6) were used for these studies. Incubation of LM6 and LM6-#6 with 10 micro M 4-HC increased the expression of FasL mRNA (2.5- and 3.0-fold, respectively). By contrast, 4-HDC, Adriamycin (ADR), cisplatin (CDP), and methotrexate (MTX) had no effect on FasL mRNA expression. Increased FasL expression after treatment with 4-HC was also demonstrated by immunohistochemistry and flow cytometry. Drug-induced FasL was functional and mediated cell death. We examined the effect of FasL up-regulation by 4-HC on LM6 and LM6-#6 cells. Flow cytometry showed that LM6-#6 cells expressed 2.2-fold more Fas than LM6 cells. Cytotoxicity of 4-HC, 4-HDC, ADR, CDP, and MTX on LM6, LM6-neo, and LM6-#6 were quantified. Colony-forming assay revealed an IC(50) of 2.10 micro M for 4-HC in LM6-neo cells compared with 0.41 micro M in LM6-#6 cells. The IC(50) for 4-HDC, ADR, CDP, and MTX were not significantly different between the two cell lines. We concluded that the increased expression of Fas enhanced LM6-#6 sensitivity to 4-HC. These data indicate that Fas/FasL may be involved in the cytotoxic pathway of CY. Combining biological agents with chemotherapeutic agents that have complementary Fas/FasL pathway actions may offer new therapeutic alternatives.     

Increased Fas expression reduces the metastatic potential of human osteosarcoma cells.

PURPOSE: The process of metastasis requires the single tumor cell that seeds the metastatic clone to complete a complex series of steps. Identifying factors responsible for these steps is essential in developing and improving targeted therapy for metastasis. Resistance to receptor-mediated cell death, such as the Fas/Fas ligand pathway, is one mechanism commonly exploited by metastatic cell populations. EXPERIMENTAL DESIGN AND RESULTS: LM7, a subline of the SAOS human osteosarcoma cell line with low Fas expression, was selected for its high metastatic potential in an experimental nude mouse model. When transfected with the full-length Fas gene (LM7-Fas), these cells expressed higher levels of Fas than the parental LM7 cells or LM7-neo control-transfected cells. These cells were also more sensitive to Fas-induced cell death than controls. When injected intravenously into nude mice, the LM7-Fas cell line produced a significantly lower incidence of tumor nodules than control cell lines. Lung weight and tumor nodule size were also decreased in those mice injected with LM7-Fas. Levels of Fas were quantified in osteosarcoma lung nodules from 17 patients. Eight samples were Fas negative, whereas the remaining 9 were only weakly positive compared with normal human liver (positive control). CONCLUSIONS: Our results demonstrate that altering Fas expression can impact the metastatic potential of osteosarcoma cells. We conclude that the increase of Fas on the surface of the LM7 osteosarcoma cells increased their sensitivity to Fas-induced cell death in the microenvironment of the lung, where Fas ligand is constitutively expressed. Thus, loss of Fas expression is one mechanism by which osteosarcoma cells may evade host resistance mechanisms in the lung, increasing metastatic potential. Fas may therefore be a new therapeutic target for osteosarcoma.     

地高辛标记探针检测重组双功能水蛭素中的DNA残留量

为检测注射用重组双功能水蛭素半成品中的DNA残留量，应用地高辛标记重组双功能水蛭素工程菌DNA，并以此为DNA探针进行点杂交，同时做特异性及灵敏度实验，结果表明，该方法检测最低限值可达1pg,且重复性好，特异性强，操作较简便，可用于双功能水蛭素制备工艺的质控及半成品的检定。