双波长薄层扫描法测定血浆中硫酸奎尼丁浓度

本文报道用薄层扫描法测定血浆中硫酸奎丁尼浓度,方法简便灵敏。血浆样品用氯仿提取后,直接点于硅胶GF_(254)板上,以环己烷:氯仿:二乙胺(5∶4∶2)为展开剂,斑点于岛津CS-930扫描仪在λs=237nm,λ_R=257nm处进行扫描。在0.4μg～8.0μg之间,线性关系良好,平均回收率为98.78%。为临床监测硫酸奎尼丁的血药浓度及动力学研究提供了可行方法。     

Analysis of ganciclovir and its related substances using high performance liquid chromatography and liquid chromatography-mass spectrometry methods

Objective High performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC/MS) methods were developed for the determination of ganciclovir and its related substances. Methods A Hypersil ODS2 column (4.6mm×250mm, 5μm) was used with a mobile phase of 0.02M potassium 1.0mL/min, and UV detector set at 254nm was used for monitoring the eluents. Results The method was simple, rapid, selective and capable of separating all related substances at trace level with a detection limit of 0.04μg/mL. It has been validated with respect to accuracy, precision, linearity, and limits of detection and quantification. The linearity range was 10.2-153.0μg/mL with r=0.9998. The percentage recoveries ranged from 96.7% to 101.6%, and RSD was 1.24%-1.96% (n=5). Conclusion The method was found to be suitable not only for monitoring the reactions during the process development but also for quality control of ganciclovir. For identification of related substances, LC/MS was used. The mainly related substances of ganciclovir active pharmaceutical ingredients (API) were determined as guanine, (1, 3-dioxolan-4-yl) methyl acetate, and diacetyl guanine.     

HPLC法测定健宝胶囊中淫羊藿苷含量

目的　建立健宝胶囊中淫羊藿苷的含量测定方法。方法　采用高效液相色谱法 ,色谱柱为C18(15 0mm×4.6mm) ,流动相为乙腈 0 .0 75mol·L-1磷酸溶液 (2 5∶75 ) ,流速 1.0mL·min-1,检测波长为 2 70nm。结果　淫羊藿苷进样量在 0 .0 6～ 0 .30 μm范围内 ,线性关系良好 (r=0 .9991) ,方法回收率为 98.5 0 %(n =5 ,RSD =1.2 0 %)。结论　本方法准确、简便、快速 ,适合于该药的质量分析检验。     

~(125)Ⅰ—联结法标记人超氧化物歧化酶

~(125)I-人超氧化物歧化酶是采用联结法进行标记,先标记羟苯丙酸酯,(Bolton Hunter Reagent),再将~(125)I-B.H.R 与SOD 相联。~(125)I-B.H.R 标记率为95%,放射性比度可达0.376 mci/μg,稳定性在两个月以上。~(125)I-SOD 联结标记率为50%,放射性比度为3.04μci/μg,用醋酸纤维膜和聚丙烯酰胺凝胶电泳后进行放射性层析扫描,放化纯在95%以上,标记不影响该酶的生物活性和免疫活性,冷冻干燥,-20℃保存两个月不失去免疫活性,放化纯仍在95%以上。     

Simultaneous determination of 12, 13-dihydroxyeuparin and glycyrrhizic acid in Yanyanfang mixture by high performance liquid chromatography

A reversed phase high performance liquid chromatography （HPLC） method was established for the simultaneous determination of 12, 13-dihydroxyeuparin and glycyrrhizic acid in Yanyanfang mixture. A Grace Apollo Cl8 column （250 mm × 4.6 mm, 5 μm） was used as the stationary phase and the mobile phase was composed of acetonitrile and aqueous phosphoric acid （0.2%, v/v）. Gradient elution was carried out at the flow rate of 1.0 mL/min and the column temperature was 30 ℃. An ultraviolet （UV） detector was used with a selected wavelength of 240 nm. Calibration curves were linear within the concentration range of 4.6-45.75 μg/mL for 12, 13-dihydroxyeuparin （r〉0.9999） and 106.9-1068.9μg/mL for glycyrrhizic acid （r〉0.9999）, respectively. Recoveries were 102.18% for 12, 13-dihydroxyeuparin and 101.17% for glycyrrhizic acid. The method developed could be applied to the simultaneous determination of 12, 13- dihydroxyeuparin and glycyrrhizic acid in Yanyanfang mixture.     

Analysis and determination of diterpenoids in unprocessed and processed Euphorbia lathyris seeds by HPLC-ESI-MS

Euphorbia lathyris （Caper spurge） is a toxic and potent Chinese materia medica （T/PCMM）. This study sought a method for identifying five diterpenoids （Euphorbia factors LI-L3, L7a, and Ls） with the spectra of UV and mass, quantifying three diterpenoids L1, L2, and L8 in crude extracts of unprocessed and processed E. lathyris seeds by liquid chromatography/ electrospray ionization mass spectrometry （LC-ESI-MS）. The analysis was achieved on an Agilent Eclipse XDB-C18 column （4.6 mm× 150mm i.d., 5 μm） with an isocratic elution with a mobile phase consisting of water and acetonitrile at a flow rate of 0.25 mL/min at column temperature of 30 ℃ and UV detection was set at 272 nm. An ESI source was used with a positive ionization mode. The calibration curve was linear in the ranges of 9.9-79 μg/mL for Euphorbia factor Lb 3.8-30.5μg/mL for Euphorbia factor L2, and 1.0-20.6 μg/mL for Euphorbia factor LB. The average recoveries （n=6） of three diterpenoids were 98.39%, 91.10% and 96.94%, respectively, with RSD of 2.5%, 2.4% and 2.1%, respectively. The contents of the three diterpenoids in processed E. lathyris seeds were 3.435, 1.367 and 0.286 mg/g, respectively, which decreased more sharply than those in unprocessed E. lathyris seeds which were 4.915, 1.944 and 0.425 mg/g, respectively. The method is simple, accurate, reliable and reproducible, and it can be applied to control the quality of unprocessed and processed E. lathyris seeds.     

甲状腺细胞的~3H-TdR摄取量测定的细胞因素影响

本文用TSH刺激FRTLS鼠甲状腺细胞,观察~3H-TdR掺入DNA的量,表达为每微克DNA内的~3H的每分钟放射计数(CPM/μgDNA)。细胞对刺激物的反应能力表达为刺激组细胞的~3H-TdR摄取值CPM/μgDNA除以对照组的CPM/μgDNA所得的“%的对照值”。-180℃冷冻2个月复苏的细胞对TSH200和2000μu/ml的反应能力分别为2741%和3028%,作为对比组的连续80天TSH培养的细胞反应能力分别为736%和719%,两组间存在统计学的显著差异。这提示用长期冷冻法可使因长期TSH培养而降低反应能力的细胞恢复良好的反应性。实验所需细胞数量应用一次种植、不经TSH增殖培养法,亦可提高细胞的反应性。     

ANTITUMOR MECHANISM OF GEM10 BY THE NATURAL KILLER ACTIVITY AND INTERLEUKIN-2 PRODUCTION

Studiesongermaniumcompoundsshowedthattheyhavemanybiologicaleffects ,suchasanti tu mor[1] ,improvingimmunity[2 ] ,reducingthecon centrationofbloodcholestrolandtrilycerides[3] ,in hibitingvirusreproduction ,butthesideeffectsoftoxicrestricttheirclinicaluses[4 ,5] .Therefore ,itisstillnecessarytodevelopothernewcompoundsoforganogermaniums.GeM 10 ,aneworganicgerma niumcompounds alkili metalcomplexcombinedwitheffectiveextractsofsomeChinesetraditionalherbs ,hasbeendemonstratedtobesaferandmorereliable.…     

Oleanolic acid-induced apoptosis and its relation with intracellular calcium in human lung adenocarcinoma A549 cells

Objective To investigate the effect of oleanolic acid (OA) on apoptosis, correlation between apoptosis and intracellular calcium, and its mechanism in human lung adenocarcinoma cell line A549. Methods Human lung adenocarcinoma A549 cells were incubated in vitro and assigned with OA concentrations of 0, 10, 20 and 40μg/mL. The apoptosis status of A549 cell line was detected with Annexin V-FITC/PI by flow cytometry (FCM); fluorescence intensity (FI) of A549 cells was assessed and the level of intracellular calcium was calculated at 24 hour of OA intervention. The relation between apoptosis and calcium FI was illustrated by curve fitting. Results FCM showed that 10, 20 and 40μg/mL of OA could induce A549 cell apoptosis, which followed a concentration-effect pattern; 24-hour intervention with 20μg/mL and 40μg/mL OA showed increased A549 cell apoptosis, and was significantly different from that with 0μg/mL OA (P<0.01). The FI of intracellular calcium concentration in 10, 20 and 40μg/mL OA groups was significantly higher than that in 0μg/mL group after 24 hours' intervention, and the FI showed a trend of increase with increased OA concentration (P<0.01). Curve fitting showed a significant correlation between apoptosis rate and intracellular calcium concentration in A549 cells (r=0.981, P<0.01). Regression equation was Y=0.508X-1.627. Conclusion OA plays a role in inducing apoptosis of human lung adenocarcinoma cells in a concentration-dependent manner. The OA-induced apoptosis is responsible for intracellular calcium overload of the tumor.     

正常SD大鼠血清及组织内源性哇巴因的含量及其意义

探讨正常 SD大鼠血清及组织内源性哇巴因 ( EO)的含量 ,为 EO生理及病理作用的进一步研究提供理论及实验依据。方法　制备高度特异性的哇巴因抗体 ,应用酶联免疫吸附试验 ( ELISA)的方法 ,对正常 SD大鼠血清及组织标本测定其 EO含量。结果　血清、肝脏、肾脏、肾上腺、垂体及下丘脑内 EO含量分别为 ( 1 .1 2± 0 .1 7) μg/ L、( 1 .79± 0 .44) μg/ kg组织、( 1 .0 8±0 .63)μg/ kg组织、( 1 .73± 0 .81 )μg/ kg组织、( 2 7.54± 6.91 )μg/ kg组织、( 1 .83± 0 .54)μg/ kg组织、( 1 0 .1 0± 3.0 8) μg/ kg组织。肾上腺内 EO含量明显高于其它组织及血清 EO含量。结论　确定了正常 SD大鼠血清及多种组织内源性哇巴因的含量 ,提示肾上腺有可能是 EO的主要来源     

应用不同血管活性药物阴茎海绵体注射排除血管性阳萎

采用不同血管活性药物及不同剂量对670例阳萎患者进行阴望海绵体内注射以排除血管性阳萎。结果96.9％的非血管性阳萎患者在应用婴栗碱30mg加酚妥拉明1mg注射后可获充分勃起，2％在罂桑碱加至60mg时获充分勃起。而对罂染减注射反应不佳的非血管性阳萎患者行PGE25μg注射后有0．9％获充分勃起，PGE1加至50μg时又有0．2％获得勃起。应用罂粟碱的患者17．8％出现异常勃起，而应用PGEI的患者46．1％出现明茎疼痛。提示不应以某种单一剂量或药物注射来排除血管性阳萎，并宜从小剂量用起，减少共发症。     

治带片中苦参碱及氧化苦参碱的HPLC法测定

目的建立治带片中苦参碱及氧化苦参碱的高效液相色谱(HPLC)测定方法。方法色谱柱:Lichrospher-NH2(4.6 mm×250 mm,5μm),C18保护柱;流动相:乙腈-无水乙醇-0.5 mol/L磷酸水溶液(80∶10∶10);检测波长212 nm;流速1.0 mL/min;柱温:室温;进样量20μL。结果苦参碱和氧化苦参碱线性范围均为1.0-10.0μg/mL,回归方程苦参碱为:C=1.201×10-4A+0.161,r=0.9992;氧化苦参碱为:C=1.366×10-4A+0.221,r=0.9996,平均回收率分别为99.9%和99.4%,RSD分别为1.48%和4.33%。结论本法简便快捷,结果准确,可用于该制剂的质量控制。     

不同性别的原发性膜性肾病的临床及病理分析

目的了解不同性别的原发性膜性肾病的临床、病理特点及其有无区别。方法回顾分析我院482例原发性膜性肾病的临床及病理资料。结果①发病年龄:男(46.89±16.73)岁,女(48.39±15.45)岁,两组差异无统计学意义(P>0.05);②临床表现:在病程、血尿、高血压的发生率方面两组无差异(P>0.05),而浮肿的发生率男性81.43%,女性88.57%,女性高于男性(P<0.05);临床主要表现为肾病综合征,男、女性分别为82.41%(253/307)、82.29%(144/175),两组无差异(P>0.05);③实验室检查:尿蛋白定量男性(4.79±2.88)g/(1.73m2.d),女性(4.15±1.85)g/(1.73m2.d),血浆白蛋白男性(25.86±7.87)g/L,女性(27.05±6.57)g/L,血IgG水平男性(7.11±3.66)mmol/L,女性(8.05±3.33)mmol/L,分别行两组间比较均有差异(P<0.05);尿蛋白定量与血浆白蛋白、血IgG呈负相关,与胆固醇呈正相关(P<0.05),男女间结果一致;尿素氮男性(6.67±4.21)mmol/L,女性(5.43±2.96)mmol/L,肌酐男性(102.82±110.3)μmol/L,女性(83.06±89.61)μmol/L,两组均有差异(P<0.05),但经体表面积校正后两组肌酐清除率比较无差异(P>0.05);④病理方面:免疫荧光检查除无肾小球的外,均为免疫复合物性肾小球肾炎,以IgG沉积为主,男性占98.96%,女性99.40%,病理类型以Ⅰ期膜性肾病最常见,男女性分别为62.54%、61.14%,两组间无差异;⑤危险度评价,男性较女性高。结论①原发性膜性肾病好发于中老年,男性多于女性;②临床主要表现为肾病综合征;③男性的尿蛋白量大于女性,而血浆白蛋白、血IgG低于女性,胆固醇高于女性;④病理以Ⅰ期膜性肾病常见。     

ICP-AES法测定血硼及儿童血硼质量浓度调查

目的探索电感耦合等离子体原子发射光谱(ICP-AES)法测定血硼的方法,确定南京地区儿童血硼质量浓度正常值范围。方法全血经1 mol/L HNO3处理后用ICP-AES法测定,并用该方法测定南京地区1 032名健康儿童及1 364名就诊儿童血硼。结果方法精密度为1.60%-4.31%,回收率为93.3%-98.9%。1 032名健康儿童全血硼质量浓度为(41.8±16.7)μg/L,1 364名就诊儿童血硼质量浓度为25.1-58.5μg/L(77%)。结论全血经硝酸处理后测定血硼,精密度及回收率均较理想,可以作为测定血硼的参考方法。并初步提出该方法的南京地区儿童血硼参考范围为25.1-58.5μg/L(1岁以下儿童除外)。     

大脑皮层下轴突纤维横断模型的建立

目的　建立一种机械性大脑皮层下轴突纤维横断动物模型。方法　对 1 0例正常 2 1 0～2 4 0 g雄性SD大鼠的大脑皮层厚度和第V层神经元深度进行测定。根据测定结果决定三组不同的横切深度 (每组用SD大鼠 8只 )。在立体定位下进行脑皮层下轴突纤维横断 ,观察动物术后肢体功能恢复情况。 1 7d后取脑进行切片和染色 ,对横切深度进行测量 ,并对神经元和神经纤维状况进行观察。结果　正常 2 1 0～ 2 4 0g雄性SD大鼠大脑皮层厚度为 ( 1 85± 0 1 2 )mm ;第V层神经元深度为 ( 1 1 7± 0 0 5 )mm。对同一预定深度所获的实际深度之间最大相差 1 60 μm。三个损伤组实际深度与预定深度差值的绝对值 |d|分别为 ( 5 0 1 7± 2 6 2 0 ) μm、( 4 6 68± 32 84)μm和 ( 4 8 70± 2 9 76) μm ,最大 |d|值为 90 μm。在损伤深度分别为 2 0 0 μm、40 0 μm和 60 0 μm时 ,神经元存活率分别为 ( 4 2 5 5± 5 1 0 ) %、( 68 96± 5 2 1 ) %和 ( 74 5 2± 5 37) %。结论　该模型模拟了多种脑白质病变导致轴突损伤的共同特征 ,重复性好 ,是进行脑轴突损伤后神经修复与再生研究较理想的模型。     

P53基因产物在肝细胞癌中的表达及其临床意义

目的 研究P53基因产物在肝细胞癌中的表达及与肝细胞癌临床病理学特征的关系。方法 应用免疫组织化学技术,检测50例肝癌及癌旁组织中P53蛋白的表达。结果 肝癌突变型P53蛋白表达阳性率为32%(16/50),癌旁组织均为阴性;有门静脉癌栓31例,P53蛋白阳性率为45%(14/31),而无门静脉癌栓19例仅2例为阳性(P＜0..05);血清甲胎蛋白(AFP)≥400μg/L 36例,P53蛋白阳性率为41%(15/36),AFP＜400μg/L 14例仅1例阳性(P＜0..05)。结论 P53基因的突变发生在肝癌启动阶段之后;P53基因突变与AFP关系密切;P53蛋白对判断肝癌预后有重要意义。     

陕西省部分地区蔬菜与水果中有机磷农药残留的调查报告

目的　了解目前陕西省部分地区蔬菜与水果食品中有机磷农药残留量的整体水平,寻求相应的预防措施。方法　采用随机与定点抽样的方法采集我省部分地区蔬菜与水果140份食品样品,通过气相色谱法测定其中8种有机磷农药残留。结果　蔬菜与水果食品中有机磷农药的检出率分别为20%和1.7%,其中蔬菜样品中对硫磷和水果样品中乐果的平均残留量分别为6.22μg/kg和3.02μg/kg,大大超过国家的限量标准。结论　陕西省存在着有机磷农药使用的不科学性,应加强管理,开展食品中农药残留的快速、常规监测,将农药残留量高的食品杜绝在进入销售市场之前。     

INHIBITION AND RADIATION SENSITIZING EFFECT OF INDOLEACETIC ACID COMBINED WITH HORSERADISH PEROXIDASE ON HELA CELLS

Indoleaceticacid (IAA )isagrowthhormoneforplants.ResearchinrecentyearshasshownthatIAApresentsacytotoxiceffectonhumancancersaf teritisoxidizedbyhorseradishperoxidase (HRP) .Becauseoflowtoxicityandstrongeffectonanaero biccells ,ithasattractedpeople sattentionasapro drugfortargettreatmentofcancers .ItsmechanismmightberelatedwiththefactthatIAAisoxidizedinthecellsofthelivingbodytoproducefreeradi cals.Theanaerobiccellgroupsincancersareoneofthemaincausesforradiationresistance .Therehavebeenfewr…     

Detection of captopril based on its enhanced resonance light scattering signals of fluorosurfactant-capped gold nanoparticles

In this study, based on its enhancement effect on resonance light scattering (RLS) of fluorosurfactant (FSN)-capped gold nanoparticles (GNPs), we reported a simple approach for the rapid sensing of captopril. Under optimum conditions, the lowest detectable concentration of captopril through this approach (S/N=3) was 0.01μg/mL. The calibration curve was linear over the range of 0.08-4.0μg/mL for the detection of captopril. The recoveries of captopril were found to fall in the range between 99% and 100%. We have validated the applicability of our method through the analyses of captopril in pharmaceutical formulations. Good agreements were obtained for the determination of captopril between the present approach and official method.     

INTRAUTERINE GROWTH RETARDATION AT FULL TERM PREGNANCIES WITH ENDOCRINE FACTORS

Intrauterinegrowthretardation(IUGR)isoneofthecriticalcomplicationsduringpregnancyandremainsthemajorcauseofperinatalmorbidityandmorta1ity.Despiteadvancesintheresearchofeti-ology,pathogenesis,diagnosis,andmanagementforIUGR,doctorscouldneitherdiagnoseitcor-rectlynormanageitefficiently.ThedataprovidedbynationalresearchinggroupforbirthdefectsshowedthattheincidencerateofIUGRwas6.38%inChina.Amongthetotalperinataldeathofneonates,themortalityofIUGRaccountsfor42.3%.Moreover,evenaftertheirbirth,th…