Flow-injection-enhanced chemiluminescence method for the determination of three β-blockers

Objective To develop a rapid, simple and sensitive chemiluminescence method for the determination of three β-blockers (bisoprolol, atenolol and propranolol). Methods The chemiluminescence of cerium (Ⅳ)-sulfite system was obviously sensitized by adding anyone of three β-blockers in acid media. A new chemiluminescence method was set up by combining with flow-injection technique and used to determine the three β-blockers. Results Good linear ranges were obtained at the concentrations of 2.0×10-7g/mL-4.0×10-5g/mL, 1.0×10-7g/mL-3.0×10-5g/mL and 7.0×10-7g/mL-1.0×10-5g/mL, respectively, with the detection limits of 5.0×10-8g/mL, 7.0×10-8g/mL and 5.0×10-8g/mL (S/N=3), respectively, and the relative standard deviations for 11 times consecutive injections of 1.0×10-6g/mL bisoprolol, atenolol and propranolol were 3.57%, 2.21% and 2.26%, respectively. Conclusion The developed method is sensitive, accurate, rapid and of low cost. And it can be applied to determine bisoprolol, atenolol and propranolol in pharmaceutical preparations.     

Simultaneous determination of berberine, matrine and oxymatrine in traditional Chinese medicines by using nonaqueous capillary electrophoresis

A rapid method for the simultaneous determination of berberine (BBR), matrine (MT) and oxymatrine (OMT) by nonaqueous capillary electrophoresis (NACE) was developed. Optimum separation of the analytes was obtained on a 50cm×50μm i.d. fused-silica capillary using a non-aqueous buffer system of 70mM ammonium acetate, 7.0% acetic acid and 10% acetonitrile at 25kV and 20℃. The relative standard deviations (R.S.D.) of the migration times and peak areas of the three active components were 0.06%-0.20% and 0.12%-3.41% for berberine, 0.11%-0.60% and 0.74%-1.63% for matrine, 0.15% and 0.45% for oxymatrine, respectively. Detection limits of berberine, matrine and oxymtrine were 0.18μg/mL, 4.08μg/mL and 4.16μg/mL, respectively. In the tested concentration range, good linear relationships (0.9992 for berberine, 0.9988 for matrine and 0.9988 for oxymatrine) were observed. The linear calibration ranges were 0.45-360.0μg/mL for berberine, 8.16-408.0μg/mL for matrine and 20.8-416.0μg/mL for oxymatrine. This method has been successfully applied to the phytochemical analysis of alkaloids extracts from two commonly used traditional Chinese herbal drugs: Sophora flavescens Ait. (Kushen) and Cortex phellodendri chinensis (Huangbai) and their medicinal preparations.     

D-山梨醇清除率法评估肝储备功能

目的探讨D-山梨醇肝脏清除率(CLh-s)法补充吲哚箐绿15min潴留率(ICGR15)和Child-Pugh分级相结合评估肝硬化门脉高压患者肝储备功能的可行性。方法41名肝硬化门脉高压患者分别给予Child-Pugh分级、ICGR15、CLh-s检测评估肝储备功能,对前两种评估方法出现交叉重叠的患者,进一步分析其CLh-s值与手术并发症的相关性。结果Child-Pugh分级与ICGR15值(以40%为界)交错重叠主要集中在B级和C级之间。在Child-Pugh分级B级的18例患者中,>40%的患者有5例,术后3例出现明显并发症(CLh-s分别为629.52、509.79、450.71 mL/min),2例未出现明显并发症(CLh-s分别为784.49、779.36 mL/min);在Child-Pugh分级C级的13例患者中,有4例ICGR15<40%的患者,2例行手术治疗,其中1例出现并发症(CLh-s为484.57 mL/min)。1例未出现明显并发症(CLh-s为723.36 mL/min)。概括之,CLh-s>700 mL/min者,手术并发症少;CLh-s<700 mL/min,者手术并发症多。结论CLh-s与肝硬化门脉高压患者手术并发症有关。该法可对ICGR15和Child-Pugh分级联合评估肝储备功能时出现交叉重叠者进一步分类,使肝储备功能评估更为准确可靠。     

四味芳香开窍药的挥发性成分对缺血缺氧PC12细胞及细胞内Ca2+的影响

目的研究四味芳香开窍药的挥发性成分对连二亚硫酸钠(Na2S2O4)致PC12细胞缺血缺氧损伤及细胞内Ca2+的影响。方法体外实验采用5mmol/L的Na2S2O4导致PC12细胞缺血缺氧性损伤,然后观测细胞形态;用二甲基噻唑二苯基四唑溴盐(MTT)比色法,考察水蒸汽蒸馏法提取的麝香、苏合香、安息香挥发性成分及冰片对缺血缺氧PC12细胞活性的影响;并按照试剂盒要求测定模型细胞内Ca2+的含量。结果麝香4、8、16μg/mL,冰片8、16μg/mL,苏合香20、40μg/mL及安息香16μg/mL组均能显著升高模型细胞的吸光度(A)值(P<0.05,P<0.01),显著抑制Na2S2O4对PC12细胞的损伤;苏合香40、80μg/mL,安息香4、8、16μg/mL组均能显著降低模型细胞内Ca2+含量(P<0.05,P<0.01);麝香、冰片有降低模型细胞内Ca2+含量的趋势。结论麝香、苏合香、安息香的挥发性成分及冰片均有保护缺血缺氧PC12细胞的作用,其机制可能与降低细胞内Ca2+的含量,防止钙超载有关。     

Capillary electrophoresis coupled with electrochemiluminescence for determination of cloperastine hydrochloride

Objective To investigate the electrochemiluminescence (ECL) behavior of cloperustine hydrochloride.Methods ECL intensity of tris (2,2'-bipyridyl) rutheniumo(Ⅱ) was enhanced, the method for the determination of dupernstine hydrochloride was established using capillary electrophoresis (CE) coupled with electrochemilumolinescence (ECL) detection. Results Under the optimum conditions, ECL intensity varied linearly with cloperastine hydrochloride concentration from 7.0×10-6 g/mL to 1.0×10-4 g/mL. The detection limit (S/N=3) was 8.05×10-7g/mL. he relative standard deviation of the ECL intensity and the migration time for 11 consecutive injections of 1.0 ×10-5g/mL cloperastine hydrochloride was 2.9% and 1.5%, respectively. This method was successfully applied to eloperastine hydrochloride tablet determination. Conclusion The method has been established, validated and applied for determination of cloperastine hydrochloride.     

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.     

流动注射-化学发光法测定氢化可的松

目的确定以高锰酸钾-亚硫酸钠体系测定氢化可的松的流动注射-化学发光分析方法。方法在酸性条件下,氢化可的松对高锰酸钾-亚硫酸钠体系发光反应具有明显的增敏作用。据此,建立了流动注射化学发光测定氢化可的松的分析方法。结果在优化的实验条件下,氢化可的松质量浓度在1.0×10-9-1.0×10-6g/mL范围内与发光强度呈良好的线性关系,检出限(3R)为4.0×10-10g/mL,对氢化可的松进行11次平行测定,其相对标准偏差为2.2%。结论本方法应用于注射液中氢化可的松含量的测定,快速、准确、简便,灵敏度高、线性范围宽。     

A rapid method for the determination of dopamine in porcine muscle by pre-column derivatization and HPLC with fluorescence detection

A rapid method has been developed based on the sample preparation procedure named as QuEChERS （Quick, Easy, Cheap, Effective, Rugged and Safe）, combined with reversed-phase high performance liquid chromatography with fluorescence detector and C18 column after precolumn derivatization using o-phthalaldehyde and 2-mercaptoethanol to determine dopamine in porcine muscle. Methanol and deionized water （0.1% acetic acid, v/v） with a ratio of 60：40 was used as mobile phase. The flow rate was 0.8 mL/min and dopamine was eluted within 15 min. The linearity range was 0.003-8 μg/mL with r=0.9992. The detection limit for dopamine was 4 μg/kg and the quantification limit was 9 μg/kg. Recovery studies were carried out at 0.1, 0.5 and 1.0 mg/kg fortification levels and the average recoveries obtained ranged from 90.4% to 98.2% with relative standard deviations between 3.5% and 8.1%. The method was found to be suitable for detection of dopamine in animal product tissues at the maximum residue level.     

Luminol-potassium permanganate chemiluminescence system for the determination of three anthracycline antibiotics

Objective To establish a flow-injection chemiluminescence method for the determination of doxorubicin, epirubicin and mitoxantrone and study its reaction mechanism. Methods In alkaline medium, chemiluminescence of luminol-potassium permanganate system could be inhibited obviously by anthracycline antibiotics. Combined with flow-injection technique, a new chemiluminescence method for determining the anthracycline antibiotics was set up. The chemiluminescence mechanism of the luminol-potassium permanganate system was also discussed. Results Under optimal conditions, the good linear ranges of doxorubicin, epirubicin and mitoxantrone were 5.0×10-9-1.0×10-6g/mL, 1.0×10-9-1.0×10-5g/mL and 3×10-9-1.0×10-6g/mL, respectively. The detection limits of doxorubicin, epirubicin and mitoxantrone were 3.0×10-9g/mL, 5.0×10-8g/mL and 2.0×10-9g/mL, respectively. During eleven repeated inter-day and intra-day precision tests of 1.0×10-6g/mL samples, the relative standard deviations corresponded to reference values of 3.0%, 2.8% and 2.1%. Conclusion The developed method is sensitive, accurate, rapid and of low cost. It can be applied to determine doxorubicin hydrochloride, epirubicin hydrochloride and mitoxantrone hydrochloride in injection preparations.     

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.     

几种静脉全麻药麻醉中知晓及遗忘作用的临床比较

目的　对比观察异丙酚、乙咪酯、硫喷妥钠三种静脉全麻药麻醉中知晓及麻醉遗忘作用 ,以寻求更适合腹腔镜胆囊切除术快速连台要求的速短效麻醉方法。方法　同期腹腔镜胆囊切除术32 5例 ,随机分为三组 ,分别用异丙酚 2mg·kg 1 、乙咪酯 0 4mg·kg 1 、硫喷妥钠 5mg·kg 1 诱导麻醉。其他诱导药物相同 (有东莨菪碱 0 .3mg、芬太尼 5 μg·kg 1 、琥珀胆碱 1 .5mg·kg 1 )。麻醉维持用 1 %普鲁卡因和 0 .0 8%琥珀胆碱复合液。术后 48h内双盲法收集麻醉中知晓资料。结果　三组知晓分别为 1 4%、5 .6%、1 3.5 % (P <0 .0 5 ) ,无痛知晓异丙酚优于其他二种 (P <0 .0 5 ) ,术毕下台知晓、离室知晓、离室清醒例数异丙酚高于另二种 (P <0 .0 5 ) ,术中作梦发生率分别为 7.9%、3.7%、4.8%(P >0 .0 5 ) ,拔管时间及离室时间异丙酚最快 (P <0 .0 5 ) ,硫喷妥钠与乙咪酯差别不大。结论　异丙酚麻醉作用时间最短 ,知晓发生率最高 ;乙咪酯麻醉作用时间长于异丙酚 ,遗忘作用最强 ;硫喷妥钠麻醉苏醒类似于乙咪酯 ,术中知晓与异丙酚相似。应根据手术 ,有针对性的选择用药。     

尿NMP22筛检复发性膀胱癌临床意义的研究

目的　探讨尿NMP2 2预测膀胱癌术后患者肿瘤复发的临床意义。方法　膀胱癌术后患者 5 0例 ,留晨尿检测NMP2 2水平 81人次。所有病例均行膀胱镜检查 ,并对可疑病变处取膀胱组织送病检。尿NMP2 2浓度采用双抗体酶标免疫分析法测定。结果　膀胱癌复发 19例 ,未复发 31例 ,两者尿NMP2 2中位数的 95 %可信区间分别为 13.9～ 5 0 .3U·mL- 1 和 4 .6～ 7.6U·mL- 1 ,无重叠。初次膀胱镜检和 或病理检查无肿瘤复发 36例 ,尿NMP2 2测值异常 (NMP2 2≥ 14 .0U·mL- 1 )和正常 (NMP2 2 <14 .0U·mL- 1 )者分别为 11例和 2 5例。经随诊前者膀胱癌复发 5例 (45 .5 % ) ,后者为 0 (0 2 5 ) ,两者复发百分比有显著性差异 ,相对危险性 (RR)为 12 5 0。结论　尿NMP2 2可作为膀胱癌术后预测肿瘤复发的指标 ,与膀胱镜相比 ,其具有可早期预测、简单、方便等优点。     

RP-HPLC method for the quantitation of β-Sitosterol in Elaeagnus Gon yanthes Benth

Objective To establish an RP-HPLC method for the determination of β-Sitosterol in Elaeagnus Gonyanthes Benth. Methods The separation was performed on a luna C8 (2) (150 mm×4.6 mm, 5μm) column with the mobile phase of methanol-water (88∶12, v/v) at a flow rate of 1.0 mL/min, the detection wavelength was set at 210 nm, and the temperature of the column was maintained at 35 ℃. Results The calibration curve of β-Sitosterol was linear over the concentration range of 0.075-0.375 mg/mL (r=0.9999) and the average recovery of β-Sitosterol was 96.30% with RSD of 3.60%(n=3). Conclusion The method is simple, rapid, and accurate, and can be used for the quality control of Elaeagnus Gonyanthes Benth.     

不同镇痛方法对子宫切除术后患者血糖、胰岛素和皮质醇水平的影响

目的　评价4 种常见的不同镇痛方法对子宫切除术后应激反应的抑制程度,以选择较为合理的镇痛方法。方法　根据镇痛方法的不同,将48例行子宫切除术患者随机分为4组:罗哌卡因复合芬太尼硬膜外镇痛组(Ⅰ组);芬太尼静脉镇痛组(Ⅱ组);罗哌卡因复合曲马多硬膜外镇痛组(Ⅲ组);曲马多静脉镇痛组(Ⅵ组)。观察麻醉前、手术后2、24、48、72 h血糖、胰岛素和皮质醇水平的变化。结果　①4种镇痛方法均能提供良好的镇痛效果;②在抑制术后高血糖和术后应激方面Ⅰ组和Ⅲ组好于Ⅱ组和Ⅵ组,尤以Ⅲ组为优。结论　硬膜外镇痛可以更有效的降低术后高血糖反应,抑制术后应激反应,以罗哌卡因复合曲马多组为更优。     

心肌肌钙蛋白-I在病毒性心肌炎诊断及预后判断中的应用

目的　探讨心肌肌钙蛋白 I(cTnI)在病毒性心肌炎诊断及预后判断中的应用。方法　选择病毒性心肌炎患者 85例 ,测定血清cTnI及肌酸激酶 MB同工酶 (CK MB) ,长期随访观察cTnI;采用超声心动图测定 ,观察心脏大小及心室功能变化。结果　①cTnI诊断病毒性心肌炎阳性率显著高于CK MB。②将患者按入院时cTnI水平由低至高分为 <0 .2 5 μg·L- 1 组 ,0 .2 5～<1 0 μg·L- 1 组及≥ 1 0 μg·L- 1 组 ,随着血清cTnI浓度的升高 ,左室舒张末内径 (LVDd)逐渐增大 ,射血分数 (EF)逐渐降低 ;随访期 ,随着入院时血清cTnI水平的由低至高 ,LVDd显著增大而EF显著降低 (P <0 .0 5 )。结论　①cTnI作为心肌损伤指标优于CK MB。②病毒性心肌炎患者 ,cTnI水平高低与患者心脏大小以及远期心室功能发展有关 ,cTnI增高者心室腔扩大而心室功能下降     

缺血后处理对缺血-再灌注大鼠肠黏膜的抗损伤作用

目的探讨缺血后处理对缺血-再灌注大鼠肠黏膜的抗损伤作用。方法复制SD大鼠肠缺血-再灌注损伤模型。实验分为4组(n=8):假手术组(S)、缺血-再灌注组(I/R)、缺血预处理组(IPC)、缺血后处理组(I-post)。比较各组大鼠肠黏膜丙二醛(MDA)含量、超氧化物歧化酶(SOD)和髓过氧化物酶(MPO)的活性变化;末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法(TUNEL)检测肠黏膜细胞凋亡发生率;Chius评分法观察肠黏膜的损伤情况。结果与I/R组相比,IPC组和I-post组大鼠肠黏膜MDA含量和MPO活性均显著降低,而SOD活性明显升高;再灌注后可见明显的肠黏膜细胞凋亡现象,I-post组和IPC组凋亡指数较I/R组显著下降,组织病理损伤亦明显减轻。I-post组和IPC组相比,各项指标无显著性差异。结论缺血后处理可通过抑制再灌注后氧自由基的过量产生,保护抗氧化系统,从而抑制肠黏膜细胞凋亡,减轻肠缺血-再灌注损伤。     

LC-ESI-MS/MS, a modified method for simultaneous quantification of isoflavonoids, flavonoids, alkaloids and saponins in a Chinese herbal preparation Gegen-Qinlian decoction

A sensitive and reliable liquid chromatography-electrospray ionisation-tandem mass spectrometry (LC-ESI-MS/MS) method was established to simultaneously quantitate four categories of compounds (isoflavonoids, flavonoids, alkaloids and saponins) in Gegen-Qinlian decoction (GQD). These compounds were separated by a Shiseido CAPCELL PAK C18 column with a linear gradient consisting of 0.1% (v/v) formic acid in water (A) and 0.1% (v/v) formic acid in acetonitrile (B), and delivered at a flow rate of 0.3 mL/min. All the analytes were determined by electrospray positive ionization tandem mass spectrometry in a multiple reaction monitoring (MRM) mode. Linearity, accuracy, precision, recovery and stability of the method were evaluated with the validation over the range of 4.0-538 5 ng/mL. The proposed method was applied to the analysis of a Chinese herbal preparation GQD successfully.     

高血压脑出血患者细胞免疫及IL-2、sIL-2R表达的意义

目的　研究高血压脑出血患者外周血中红细胞免疫和T淋巴细胞亚群的改变以及白细胞介素 2 (IL 2 )、可溶性白细胞介素 2受体 (sIL 2R)表达的变化规律 ,探讨它们之间相互关系及临床意义。方法　采用酶联免疫吸附 (ELISA)法测定患者IL 2、sIL 2R的含量 ,免疫粘附法测定红细胞免疫活性及其调节功能 ,S P(链亲和素 过氧化物酶 )一步法测定CD3、CD4、CD8的细胞数。结果　高血压脑出血组IL 2含量较对照组明显降低 (P <0 .0 5 ) ,sIL 2R则明显升高 (P <0 .0 5 ) ;红细胞C3b受体 (RBC .C3bR)、红细胞免疫调节促进因子 (RFER)明显降低 (P <0 .0 0 1) ,红细胞免疫复合物 (RBC .ICR)、红细胞免疫调节抑制因子 (RFIR)则明显升高 ;CD3、CD4细胞数明显降低 (P <0 .0 1) ,CD8无明显差异 (P >0 .0 5 )。结论　高血压脑出血患者出现免疫功能低下。通过临床检测患者IL 2、sIL 2R含量、红细胞免疫功能、T细胞亚群活性在高血压脑出血的发生、发展、治疗、转归及预后方面具有重要的临床指导意义     

Study of the mechanism on the apoptosis induced in Human leukemia cell line K562 by the combination of indole-3-acetic acid and horseradish peroxidase

Objective To investigate the mechanisms of apoptosis induced in Human leukemia cell line K562 by the combination of indole-3-acetic acid and horseradish peroxidase. Methods Human leukemia cell line K562 were exposed to indole-3-acetic acid （IAA） at 20, 40, 60, 80 or 100 mol/L and horseradish peroxidase（HRP） at 1.2 g/mL for varying times. MTT assay was applied to detect the cell proliferation. Flow cytometry was performed to detect the arrest of cell cycle. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling （TUNEL） assay was used to measure apoptosis. 2, 7-dichlorofluorescin diacetate （DCFH-DA） uptake was measured to determine free radical by confocal microscope. Content of malondiadehyde （MDA） and activity of superoxide dismutase （SOD） were measured by biochemical methods. Results IAA/HRP initiated growth inhibition of K562 cells in a dose- and time-dependent manner. Flow cytometry revealed that cell cycle arrested at G1/G0 after 24 hours treatment. After 72 hours treatment, apoptotic rate of 100 mol/L IAA group increased to 43.9 %, which was 5 times that of control（P〈0.01）. Content of MDA and activity of SOD increased respectively in treatments compared with control. Meanwhile, IAA/HRP stimulated the formation of free radical, which was increased by IAA concentration-dependently. Conclusion The combination of IAAand HRP can inhibit the growth of Human leukemia cell line K562 in vitro by inducing apoptosis which is associated with the increase of free radical. The combination of IAA and HRP might be a promising chemopreventive and chemotherapeutic agent against human leukemia.