白花蛇舌草总黄酮的提取及其对BGC-823胃癌细胞增殖周期与凋亡的影响

[目的]提取白花蛇舌草总黄酮,探讨白花蛇舌草总黄酮对BGC-823胃癌细胞周期和凋亡的影响。[方法]用乙醇回流法提取白花蛇舌草总黄酮,标准曲线法测定其含量,AB-8大孔吸附树脂进行纯化,然后作用于BGC-823肿瘤细胞,流式细胞仪检测FHD对细胞周期及细胞凋亡的影响。[结果]乙醇回流法提取白花蛇舌草总黄酮,含量为67.2mg/g(总黄酮/干浸膏),纯度为6.72%。AB-8大孔吸附树脂纯化白花蛇舌草总黄酮后所得精制品中总黄酮纯度约为23.82%,纯化后的总黄酮纯度提高约4倍。白花蛇舌草总黄酮组较对照组G1期细胞增加,S期细胞明显减少;实验组凋亡细胞比例明显高于对照组。[结论]AB-8大孔吸附树脂对白花蛇舌草中总黄酮具有很好的纯化富集作用;白花蛇舌草总黄酮对胃癌细胞BGC-823株的生长有明显的抑制作用,作用机制可能与抑制DNA的合成,诱导细胞凋亡有关。     

大孔吸附树脂提取纯化杜仲叶绿原酸的研究

目的 提取纯化杜仲叶绿原酸，确定最佳吸附树脂吸附-解吸条件。方法 以乙醇提取液为吸附原液，静态吸附实验筛选树脂；动态吸附实验确定最佳树脂的吸附-解吸条件。结果 NKA-Ⅱ树脂最佳吸附-解吸条件：吸附流速2ml／min，原液过柱吸附2次，最佳吸附剂为40％乙醇。结论 NKA-Ⅱ树脂是吸附绿原酸的理想吸附剂，适宜于产业化生产。     

二色补血草总黄酮的提取纯化及其对大鼠激素水平的影响

目的 以二色补血草醇提液为原料,利用大孔树脂对其总黄酮进行纯化处理,并探讨其对去势大鼠血清雌二醇(E2)及孕酮(P)的影响.方法 总黄酮的提取工艺为料液比1∶15,浓度为60％的乙醇为提取剂,于50℃水浴中提取3h,共回流提取2次.用D-101大孔吸附树脂分别用水(3倍量)和70％乙醇(5倍量)以1.5 ml/min的流速进行洗脱,合并70％乙醇洗脱液,回收乙醇,干燥后得二色补血草总黄酮浸膏.采用去势造模法建立动物模型,分为二色补血草总黄酮低、中、高剂量组、宫血宁对照组、模型对照组和假手术组.每天灌胃给药1次,连续给药14 d,末次给药后进行眼眶取血待测.结果 二色补血草醇提液浓缩至生药量0.24 g/ml,纯化后浸膏中总黄酮含量为65.46％,即每克浸膏中含总黄酮0.065 46 9.结论 给药后,二色补血草总黄酮各剂量组和宫血宁组均可提高去势大鼠血清中E2和P的含量,其中二色补血草总黄酮高剂量组对模型大鼠血清中的E2水平具有明显的改善作用,且呈现一定的量效关系.     

多定量指标研究千里香提取物工艺

目的优选千里香药材乙醇提取物最佳工艺条件。方法以千里香醇提浸膏量、浸膏中九里香酮和5'-九里香酮的含量为考察指标,采用正交实验法对影响醇提工艺的主要因素:乙醇浓度、乙醇用量、提取时间和提取次数进行研究。结果千里香乙醇提取物提取最佳工艺为:用浓度为80%的乙醇提取,乙醇用量为药材量10倍,提取两次,每次1小时。结论该提取工艺方法可行、简便、合理。     

碱化血浆紫外线蓝光照射树脂吸附清除胆红素

目的探讨用紫外线蓝光混合光照射碱化血浆再用树脂吸附对胆红素影响,建立净化人工肝分离血浆新方法。方法 （一）将人工肝分离出的血浆碱化成pH10（血浆：0.2mol/LNa2CO3=1∶1,V/V）,用紫外线蓝光照射（10～14）min,再用D4006（交联PS塑料）非极性吸附树脂流动吸附3min,检测胆红素和蛋白浓度。（二）将人工肝分离出的血浆碱化成pH10（血浆：2mol/LNa2CO3=10∶1,V/V）,用紫外线蓝光照射15min,再用D4006树脂流动吸附5min,检测胆红素和蛋白浓度。结果在方法一,碱化血浆照射组的胆红素（TBil、DBil、IBil）清除率分别为86%、84%、87.5%,Alb回收率99%;碱化血浆照射吸附组的TBil、DBil、IBil清除率均为98%,Alb回收率90%。在方法二,碱化血浆照射组的TBil、DBil、IBil清除率分别为85%、84%、86%,Alb回收率99%;碱化血浆照射吸附组的TBil,DBil,IBil清除率分别为90%、87%、91%,Alb回收率90%。结论紫外线蓝光照射碱化血浆可明显降低胆红素浓度,照射后再用D4006树脂吸附可进一步降低胆红素浓度,且Alb回收率达90%～99%。故本法有望用于人工肝血液净化。     

国产纳米炭对丝裂霉素的吸附和解吸附性能研究

目的 应用国产纳米炭(CNP)吸附丝裂霉素(MMC)制备淋巴靶向制剂.方法采用等比法设计CNP吸附MMC实验,以2n方式稀释CNP-MMC溶液设计CNP解吸附MMC实验,应用高压液相色谱法测定分离液中游离的MMC浓度,计算吸附率和解吸附率.结果 [1]吸附结果:含CNP0.25、0.5、1、2、4ml溶液中游离的MMC浓度分别为(1240±116)、(774±83)、(300±48)、(128±26)、(111±18)μg/ml,吸附率分别为94.5%、93.6%、85.0%、61.3%、38.0%.随CNP量的增加,吸附率逐渐增加(P<0.01),至4ml时吸附率与2ml时变化不大(P>0.05).[2]解吸附结果:稀释后溶液体积为4、16、32、128、512ml组对CNP-MMC解吸附后,溶液中游离MMC浓度分别为(128±26)、(122±25)、(117±25)、(50±16)、(13±2)μg/ml,解吸附率分别为6.6%、24.4%、46.8%、80.0%、83.2%(P<0.01).结论 CNP对MMC具有吸附和解吸附性能.1ml注射用水和1ml CNP溶解4mgMMC粉针剂可作为制剂的最佳吸附和解吸附配比.     

白果总黄酮超声波提取及抗衰老作用

目的探讨确立白果总黄酮最佳提取方法并研究其抗衰老活性。方法选取4个品种的白果:佛手、佛指、金坠、圆铃,采用正交法确立总黄酮最佳超声波提取工艺,并通过芦丁标准曲线测定提取物中的总黄酮含量;采用光度计比色法测定4个品种白果总黄酮提取物的清除二苯基苦基苯肼(DPPH)自由基,清除超氧阴离子自由基,清除2,2-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐(ABTS)自由基的能力以及还原能力。结果白果总黄酮超声波提取最佳工艺为:时间50 min、温度80℃、乙醇体积分数80%、料液比1∶15;4种白果总黄酮含量均较高,其中由高到低依次为:佛指金坠佛手圆铃。抗衰老作用研究显示,佛指清除DPPH自由基、超氧阴离子自由基、ABTS自由基和还原能力最强,其次为圆铃,两者均优于佛手、金坠;且4种白果总黄酮均在质量浓度400μg/ml时达到较高水平,之后进入平台期。结论白果总黄酮具有较好的抗衰老作用,其中以佛指总黄酮效果最佳。     

人用地鼠肾细胞狂犬病疫苗纯化工艺中上样量的选择

经过Ｓｅｐｈａｒｏｓｅ 4ＦＦ凝胶柱层析法〔1〕纯化的人用地鼠肾细胞狂犬病疫苗 ,其杂蛋白去除率可达99 %以上 ,副反应率较现行的 3- 5倍浓缩疫苗明显降低 ,且免疫效价可达 3.7ＩＵ /ｍｌ以上 ,值得大力推广并最终取代浓缩疫苗。在深入研究纯化工艺最佳条件时 ,发现待纯化的浓缩疫苗上样量的多少直接影响纯化后疫苗的质量 ,研究结果表明 ,上样量为柱床体积的 10 %以下时 ,纯化效果最佳。1　材料和方法1.1　细胞与病毒培养　使用 12～ 14ｇ健康金黄地鼠 ,无菌取肾 ,按常规方法制备单层细胞 ,用 3Ｌ转瓶 37℃及 33℃常规培养 ,于接种后第 4…     

鸡眼草总黄酮的提取工艺条件及其体外抗氧化性

目的 研究鸡眼草总黄酮的提取工艺条件及其体外抗氧化活性.方法 通过单因素实验和正交试验对鸡眼草总黄酮的提取工艺条件进行优化.采用清除羟自由基(·OH)、清除超氧阴离子自由基(0(2))、抗O(2)活力和总抗氧化能力(T-AOC)等方法评价其黄酮类化合物的体外抗氧化活性.结果 鸡眼草总黄酮得的最佳提取工艺条件为:取温度为70℃、乙醇浓度为55％、提取时间为2.5h、料液比为1:50.随着鸡眼草总黄酮浓度的增加,对·OH和0(2)的清除率逐渐升高,抗0(2)活力和T-AOC亦随之增强.结论 优选的鸡眼草总黄酮提取工艺合理、稳定可行,鸡眼草总黄酮具有一定的体外抗氧化活性.     

经颅彩色多普勒超声对颅内段椎动脉狭窄血流动力学的评价标准

目的探讨经颅彩色多普勒超声(TCCS)诊断颅内段椎动脉狭窄的血流动力学参数标准。方法回顾性纳入622例门诊或住院可疑后循环缺血患者,从中选取经TCCS、彩色多普勒血流显像(CDFI)筛查和DSA检查的患者共216例,其中颅内椎动脉正常者33例(15.3%),狭窄率50%者45例(20.8%),狭窄率为50%~69%者44例(20.4%),狭窄率为70%~99%者94例(43.5%)。通过检测颅内段椎动脉、椎间隙段收缩期峰值流速(PSV1、PSV2)和舒张期末流速(EDV1、EDV2),分别计算颅内段平均流速(MV)、颅内段与椎间隙段收缩期峰值流速、舒张期末流速的比值SPRP(PSV1/PSV2)、SPRE(EDV1/EDV2)。以DSA检查结果为标准,计算受试者工作特征(ROC)曲线下面积,获得最佳截断点。结果 TCCS诊断颅内段椎动脉狭窄血流动力学参数最佳截断点分别为:狭窄率50%的参数标准为110 cm/s≤PSV1≤145 cm/s,65 cm/s≤MV≤85 cm/s;狭窄率为50%~69%的参数标准为145 cm/s≤PSV1≤190 cm/s,85 cm/s≤MV≤115 cm/s;狭窄率为70%~99%的参数标准为PSV1≥190 cm/s,MV≥115 cm/s。结论 TCCS可有效评价颅内段椎动脉狭窄的血流动力学变化,为颅内段椎动脉狭窄的超声评判标准提供参考。     

妊娠时间与肺结核复发的相关性研究及诊治对策

目的 分析肺结核史患者妊娠时间和肺结核复发间相关性.方法 选取我院收治的有肺结核史的妊娠妇女576例作为研究对象,对其妊娠前肺结核治疗、治愈后妊娠时间、妊娠后复发肺结核等进行分析,总结有肺结核史育龄女性的妊娠时间和肺结核复发之间的关系.结果 肺结核治愈后不同时间段妊娠者的结核复发率比较,差异具有显著性（P＜0.05）,停药后间隔时间越久妊娠,肺结核复发的几率越小.结论 加强孕期痰菌检查,及早发现复发肺结核,提高母婴安全.     

我国骨关节结核诊治的回顾与展望

骨关节结核是危害人们健康的严重感染性疾病,近95％由他处结核病继发而来.罹患骨关节结核疾病后几乎均将致残,严重影响人们的健康、工作和生活.建国以来在党和国家的关心和支持下,骨关节结核的诊治水平取得了长足进步.时至今日,由于多种原因,学科发展和被重视程度受到一定的制约,同整个医疗行业的发展不相适应.回顾过去,展望未来,我们需要重新审视骨关节结核的诊治方法,努力推进骨关节结核诊疗技术的科学发展.     

Effect of phosphorylation of MAPK and Stat3 and expression of c-fos and c-jun proteins on hepatocarcinogenesis and their clinical significance

AIM To study the effect of phosphorylation ofMAPK and Stat3 and the expression of c-fos andc-jun proteins on hepatocellular carcinogenesisand their clinical significance.METHODS SP immunohistochemistry was usedto detect the expression of p42/44~(MAPK), p-Stat3,c-fos and c-jun proteins in 55 hepatocellularcarcinomas (HCC) and their surrounding livertissues.RESULTS The positive rates and expressionlevels of p42/44~(MAPK), p-Stat3, c-fos and c-junproteins in HCCs were significantly higher thanthose in pericarcinomatous liver tissues (PCLT).A positive correlation was observed between theexpression of p42/44~(MAPK) and c-fos proteins, andbetween p-Stat3 and c-jun, but there was nosignificant correlation between P42/44~(MAPK) and p-Stat3 in HCCs and their surrounding livertissues.CONCLUSION The abnormalities of Ras/Raf/MAPK and JAKs/ Stat3 cascade reaction maycontribute to malignant transformation ofhepatocytes. Hepatocytes which are positive forp42/ 44~(MAPK), c-fos or c-jun proteins may bepotential malignant pre-cancerous cells.Activation of MAPK and Stat3 proteins may be anearly event in hepatocellular carcinogenesis.     

Effect of phosphorylation of MAPK and Stat3 and expression of c-fas and c-jun proteins on hepatocarcinogenesis and their clinical significance

AIM To study the effect of phosphorylation ofMAPK and Stat3 and the expression of c-fos andc-jun proteins on hepatocellular carcinogenesisand their clinical significance.METHODS SP immunohistochemistry was usedto detect the expression of p42/44MAPK, p-Stat3,c-fos and c-jun proteins in 55 hepatocellularcarcinomas (HCC) and their surrounding livertissues.RESULTS The positive rates and expressionlevels of p42/44MAPK, p-Stat3, c-fos and c-junproteins in HCCs were significantly higher thanthose in pericarcinomatous liver tissues (PCLT).A positive correlation was observed between theexpression of p42/44MAPK and c-fos proteins, andbetween p-Stat3 and c-jun, but there was nosignificant correlation between p42/44MAPK and p-Stat3 in HCCs and their surrounding livertissues.CONCLUSION The abnormalities of Ras/Rat/MAPK and JAKs/ Stat3 cascade reaction maycontribute to malignant transformation ofhepatocytes. Hepatocytes which are positive forp42/ 44MAPK, c-fos or c-jun proteins may bepotential malignant pre-cancerous cells.Activation of MAPK and Stat3 proteins may be anearly event in hepatocellular carcinogenesis.     

Overweight and Obesity and Progression of ADPKD

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Replication and Inhibitors of Enteroviruses and Parechoviruses

The Enterovirus (EV) and Parechovirus genera of the picornavirus family include many important human pathogens, including poliovirus, rhinovirus, EV-A71, EV-D68, and human parechoviruses (HPeV). They cause a wide variety of diseases, ranging from a simple common cold to life-threatening diseases such as encephalitis and myocarditis. At the moment, no antiviral therapy is available against these viruses and it is not feasible to develop vaccines against all EVs and HPeVs due to the great number of serotypes. Therefore, a lot of effort is being invested in the development of antiviral drugs. Both viral proteins and host proteins essential for virus replication can be used as targets for virus inhibitors. As such, a good understanding of the complex process of virus replication is pivotal in the design of antiviral strategies goes hand in hand with a good understanding of the complex process of virus replication. In this review, we will give an overview of the current state of knowledge of EV and HPeV replication and how this can be inhibited by small-molecule inhibitors.     

Effects of sex and time of day on metabolism and excretion of corticosterone in urine and feces of mice

Non-invasive techniques to monitor stress hormones in small animals like mice offer several advantages and are highly demanded in laboratory as well as in field research. Since knowledge about the species-specific metabolism and excretion of glucocorticoids is essential to develop such a technique, we conducted radiometabolism studies in mice (Mus musculus f. domesticus, strain C57BL/6J). Each mouse was injected intraperitoneally with 740 kBq of 3H-labelled corticosterone and all voided urine and fecal samples were collected for five days. In a first experiment 16 animals (eight of each sex) received the injection at 9 a.m., while eight mice (four of each sex) were injected at 9 p.m. in a second experiment. In both experiments radioactive metabolites were recovered predominantly in the feces, although males excreted significantly higher proportions via the feces (about 73%) than females (about 53%). Peak radioactivity in the urine was detected within about 2h after injection, while in the feces peak concentrations were observed later (depending on the time of injection: about 10h postinjection in experiment 1 and about 4h postinjection in experiment 2, thus proving an effect of the time of day). The number and relative abundance of fecal [3H]corticosterone metabolites was determined by high performance liquid chromatography (HPLC). The HPLC separations revealed that corticosterone was extensively metabolized mainly to more polar substances. Regarding the types of metabolites formed, significant differences were found between males and females, but not between the experiments. Additionally, the immunoreactivity of these metabolites was assessed by screening the HPLC fractions with four enzyme immunoassays (EIA). However, only a newly established EIA for 5alpha-pregnane-3beta,11beta,21-triol-20-one (measuring corticosterone metabolites with a 5alpha-3beta,11beta-diol structure) detected several peaks of radioactive metabolites with high intensity in both sexes, while the other EIAs showed only minor immunoreactivity. Thus, our study for the first time provides substantial information about metabolism and excretion of corticosterone in urine and feces of mice and is the first demonstrating a significant impact of the animals' sex and the time of day. Based on these data it should be possible to monitor adrenocortical activity non-invasively in this species by measuring fecal corticosterone metabolites with the newly developed EIA. Since mice are extensively used in research world-wide, this could open new perspectives in various fields from ecology to behavioral endocrinology.     

血清C-反应蛋白和红细胞分布宽度在AECOPD与稳定期的变化及意义

目的观察慢性阻塞性肺疾病(COPD)患者急性加重期入院、出院时血清C-反应蛋白(CRP)和红细胞分布宽度(RDW)与正常人的差异。方法分别检测健康老年组(年龄≥65岁)、COPD急性加重期患者入院、出院时CRP和RDW水平,比较组间各指标的差异。结果急性加重期患者入院、出院时CRP和RDW水平均高于健康老年组,入院时CRP水平高于出院时(P0.01),入院、出院时RDW水平无统计学差异。结论 CRP和RDW可一定程度评估COPD病情及反映病情变化。     

心电图、心电向量图与冠状动脉造影结果对比分析

目的:通过分析心电图(Electrocardiogram,ECG)和心电向量图(Vectorcardiogram,VCG)的改变与冠脉造影（CAG）结果进行对比,探讨ECG、VCG在冠状动脉病变中的诊断价值。方法: 选择2008年1月~2009年12月临床拟诊断为冠心病患者108例,行常规ECG、VCG检查,并于1周内进行CAG,对检查结果依据各自的诊断标准进行判定,以CAG为标准诊断法,利用四格表法,计算相关评价真实性的指标并进行比较。结果: ①VCG检测的灵敏度、特异度、准确度显著高于ECG(P<0.05,P<0．01)。②ECG、VCG阳性率与冠脉病变支数组间比较:在单支病变、双支病变中,VCG阳性率明显高于ECG(P<0.05),左主干或三支病变无统计学意义;组内比较:ECG组左主干或三支病变组较单支病变、双支病变阳性率高(P<0.05,P<0.01);VCG组左主干或三支病变组较单支病变阳性率高(P<0.05);与双支病变阳性率比较无统计学意义;③ECG、VCG阳性率与冠脉病变程度组间比较:冠脉病变狭窄50%～69%的VCG阳性率明显高于ECG (P<0.05),其他两组阳性率比较无统计学意义;组内比较:ECG组冠脉病变狭窄≥90%较50%～69%、70%～89%的阳性率高(P<0.05,P<0.01); VCG组狭窄≥90%较50%～69%阳性率高（P<0.01),其他无统计学意义。结论: VCG对冠心病检测价值显著高于ECG。     

Detection and characterization of the product of hydroethidine and intracellular superoxide by HPLC and limitations of fluorescence

Here we report the structural characterization of the product formed from the reaction between hydroethidine (HE) and superoxide (O(2)(.-)). By using mass spectral and NMR techniques, the chemical structure of this product was determined as 2-hydroxyethidium (2-OH-E(+)). By using an authentic standard, we developed an HPLC approach to detect and quantitate the reaction product of HE and O(2)(.-) formed in bovine aortic endothelial cells after treatment with menadione or antimycin A to induce intracellular reactive oxygen species. Concomitantly, we used a spin trap, 5-tert-butoxycarbonyl-5-methyl-1-pyrroline N-oxide (BMPO), to detect and identify the structure of reactive oxygen species formed. BMPO trapped the O(2)(.-) that formed extracellularly and was detected as the BMPO-OH adduct during use of the EPR technique. BMPO, being cell-permeable, inhibited the intracellular formation of 2-OH-E(+). However, the intracellular BMPO spin adduct was not detected. The definitive characterization of the reaction product of O(2)(.-) with HE described here forms the basis of an unambiguous assay for intracellular detection and quantitation of O(2)(.-). Analysis of the fluorescence characteristics of ethidium (E(+)) and 2-OH-E(+) strongly suggests that the currently available fluorescence methodology is not suitable for quantitating intracellular O(2)(.-). We conclude that the HPLC/fluorescence assay using HE as a probe is more suitable [corrected] for detecting intracellular O(2)(.-).