乙醇沉淀法与壳聚糖絮凝沉降法对疏郁复方颗粒中芍药苷含量的影响及纯化效果比较

目的比较乙醇沉淀法与壳聚糖絮凝沉降法对疏郁复方颗粒提取液的精制效果。方法采用正交试验法考察药液浓度、醇浓度、静置时间对乙醇沉淀法精制疏郁复方颗粒提取液的影响,并比较乙醇沉淀法与壳聚糖絮凝沉降法对固形物与芍药苷含量的影响。结果最佳的乙醇沉降工艺为:药液浓度0.8 g原药材/mL、醇浓度20%、静置6 h。乙醇沉淀法和壳聚糖絮凝沉降法均降低了疏郁复方颗粒煎煮液的出膏率,但壳聚糖絮凝沉降法对疏郁复方颗粒中芍药苷含量造成了较大损失。结论低浓度的醇沉工艺可以用于疏郁复方颗粒提取液的纯化,壳聚糖絮凝沉降法是否适用于该提取液的精制有待于进一步研究。     

Problems with PEG-based precipitation methods in the determination of HDL2- and HDL3-cholesterol

In this study a comparison of two polyethylene glycol (PEG)-based precipitation methods, that is, one using PEG-6000/DS-15000 and the other using two different concentrations of PEG-20000 are presented. The total HDL-cholesterol values measured with the PEG-6000 procedure were higher (13%) than those measured with PEG-20000 (r=0.91) in 16 pooled serum samples tested. The HDL₂- and HDL₃-cholesterol values measured with the two methods differed markedly from each other in spite of the moderate correlation (HDL₂: r=0.74; HDL₃: r=0.26) observed. The advantage of the PEG-20000 method as compared with the other method was its simplicity and rapidity. The separation of HDL₂/HDL₃ is more or less arbitrary in both methods used and further studies are needed for the estimation of optimal conditions for subclass separation.     

A novel high-pressure precipitation tandem homogenization technology for drug nanocrystals production – a case study with ursodeoxycholic acid

Abstract

To overcome the limitations of the conventional particle size reduction technologies, a novel combinative particle size reduction method for the effective production of homogeneous nanosuspensions was investigated. Ursodeoxycholic acid, a poorly soluble drug representative, was tried to prepare nanosuspension by homogenization technology and high-pressure precipitation tandem homogenization technology. It was shown that the combinative approach could significantly improve the particle size reduction effectiveness over conventional homogenization approach. The Box–Behnken design analysis for process optimization revealed that the acceptable UDCA-NS was obtained wherein the optimal values of A, B, C and D were 10%, 500?bar, 0.125 and 600?bar, respectively. SEM results demonstrated that no significant aggregation or crystals growth could be observed in the freeze-dried UDCA nanocrystals. The DSC and XRD results showed that UDCA remained in a crystalline state. Dissolution velocities of the freeze-dried UDCA-NS powder were distinctly superior compared to those of the crude powder and physical mixture. The high-pressure precipitation tandem homogenization technology can be a good choice for nanosuspension preparation of poorly soluble UDCA, due to high efficiency of particle size reduction.     

夏枯草水提液及水提醇沉上清液降血压药效评价

目的： 探讨夏枯草水提液及水提醇沉上清液对自发性高血压大鼠（spontaneously hypertensive rat,SHR）血压的量效和时效关系。 方法： 采用水提、水提醇沉2种方法处理夏枯草,以SHR大鼠（n=72）为模型,以卡托普利作为阳性对照,考察不同剂量（2.5,5,10 g·kg^-1）的夏枯草水提液和醇沉上清液单次灌胃给药后不同时间点（0,2,5,7,10 h）对SHR 大鼠收缩压、舒张压、平均动脉压、心率等指标的影响。 结果： 给药后2 h 夏枯草水提液和醇沉上清液不同剂量均能显著降低SHR收缩压（p<0.05）,醇沉上清液高剂量（10 g·kg^-1）组能显著降低SHR舒张压（P<0.05）;给药后5 h,夏枯草水提液低剂量（2.5 g·kg^-1）组开始对SHR收缩压无显著影响,醇沉高剂量（10 g·kg^-1）组进一步降低舒张压;给药后7 h,仅醇沉上清液高剂量对SHR收缩压有显著影响（P<0.05）,其余各组对SHR舒张压、SHR平均动脉压和心率均无显著性影响。 结论： 夏枯草水提醇沉所得的上清液在去除部分杂质的情况下保留和富集了夏枯草降血压有效组分,为进一步研究夏枯草降血压物质基础及其制剂工艺提供参考。     

膝悦颗粒提取及纯化工艺优选

目的:优选膝悦颗粒的提取及纯化工艺。方法:采用HPLC-ELSD测定黄芪甲苷含量,流动相乙腈-水(35∶65);采用HPLC测定龙胆苦苷含量,流动相甲醇-水(27∶73),流速0.8 mL·min-1,检测波长271 nm。以黄芪甲苷和龙胆苦苷含量的综合评分为指标,选择加水量、提取时间、提取次数为考察因素,采用正交试验优选膝悦颗粒的提取工艺;以黄芪甲苷、龙胆苦苷含量及出膏率为评价指标,通过单因素试验筛选膝悦颗粒的纯化工艺。结果:最佳提取工艺为加10倍量水煎煮3次,每次1 h;黄芪甲苷、龙胆苦苷质量分别为4.34,43.7 mg。采用醇沉法,使含醇量达50%;黄芪甲苷21.30 mg(保留率85.61%),龙胆苦苷216.32 mg(保留率97.74%),出膏率22.13%。结论:优选的提取及纯化工艺稳定可行,为膝悦颗粒的工业化生产提供参考。     

Recruitment of ataxia-telangiectasia mutated to the p21(waf1) promoter by ZBP-89 plays a role in mucosal protection

BACKGROUND & AIMS: Histone deacetylase inhibitors (HDACi) induce growth arrest, apoptosis, and differentiation, particularly in colon cancer cells where they are potential chemopreventive agents. HDACi induction of the cyclin-dependent kinase inhibitor p21(waf1) has been shown to require ataxia-telangiectasia mutated (ATM). Nevertheless, how ATM participates in p21(waf1) gene expression has not been defined. METHODS: In vivo protein complexes forming in response to butyrate were studied using co-immunoprecipitation and mass spectroscopy. DNA elements in the p21(waf1) promoter were analyzed in vivo by chromatin immunoprecipitation and in vitro DNA affinity precipitation assays. The expression of p21(waf1) was analyzed by immunoblots and reporter assays. RESULTS: Reduction of ZBP-89 or ATM with small interfering RNAs blocked HDACi-induced p21(waf1) expression. Chromatin immunoprecipitation and DNA affinity precipitation assays showed that both ZBP-89 and ATM are recruited to the GC-rich DNA elements of the p21(waf1) promoter with HDACi treatment. Co-immunoprecipitation revealed that ATM associates with ZBP-89 in an HDACi-dependent manner. Serial deletions revealed that ATM interacts with both the N-terminal and DNA binding domains of ZBP-89. Moreover, we found that immunodepletion of ZBP-89 prevented recruitment of ATM to the p21(waf1) promoter in vitro. Silencing of ZBP-89 expression blocked HDACi-induced phosphorylation of ATM(Ser1981) and p53(Ser15). ATM(Ser1981) phosphorylation in the colons of mutant mice expressing an N-terminally truncated form of ZBP-89 was not observed after ingestion of dextran sodium sulfate and correlated with exacerbation of the mucosal injury. CONCLUSIONS: ZBP-89 interacts with ATM in a butyrate-dependent manner and is essential for colonic homeostasis in the setting of acute mucosal injury.     

Determination of berberine, palmatine and jatrorrhizine in rabbit plasma by liquid chromatography-electrospray ionization-mass spectrometry

Incurred rabbit plasmas samples were utilized for method quality assessment in this study, where an optimized protein precipitation method for the preparation of rabbit plasma samples and a rapid and sensitive liquid chromatography-electrospray ionization-mass spectrometry for the simultaneous determination of berberine, palmatine and jatrorrhizine was described. Plasma samples (100 μl) were pretreated by protein precipitation with the mixture of 3% formic acid and 50 ng/ml clozapine (internal standard) in acetonitrile followed by LC analysis using a C₁₈ column and a mobile phase composed of 0.4% formic acid solution and 0.2% formic acid solution of methanol (60:40, v/v) operated at a flow rate of 0.4 ml/min. The analysis was performed in the multiple reaction monitoring mode via electrospray ionization source operating in the positive ionization mode. The method was linear over the concentration range of 0.1-400 ng/ml for all target components. The lower limits of quantification were 0.1 ng/ml for all analytes, all intra- and inter-day precision values were less than 7.10%, and accuracy (bias, %) was within ±7.11%. The mean absolute recovery was more than 72% for all analytes. The validated method has been successfully applied to the pharmacokinetic study of berberine, palmatine and jatrorrhizine in rabbit plasma after oral administration of San-Huang decoction to rabbits.     

臭氧联合椎间孔胶原酶注射法治疗极外侧型腰椎间盘突出临床疗效观察

目的探讨治疗极外侧型腰椎间盘突出症的治疗方法及疗效。方法在C型臂监视下,对30例极外侧型腰椎间盘突出症患者进行了盘内臭氧联合患侧椎间孔处注射胶原酶治疗方法。结果术后电话随访6～12个月,按改良Macnab评判标准,优：20例,术后疼痛消失,直腿抬高试验〉70,活动无限制,良：6例,偶有非神经根性疼痛,术前体征明显改善,可从事轻体力劳动,不需服用镇痛药。可：3例,症状和体征有些改善,但仍疼痛,直腿抬高试验较前增高,但〈70°,常有疼痛,活动受一定限制,偶有服用镇痛药。差：1例,仍有神经根性疼痛,症状和体征无改善。治疗优良率87%,有效率96.6%。结论在X线监视下,对极外侧突出椎间盘内、椎小关节旁注射臭氧联合患侧椎间孔处注射胶原酶,可有效解除髓核组织对神经根的压迫,位置准确,疗效可靠,是治疗极外侧型腰椎间盘突出症的良好方法。     

HPLC法测定人血浆中吉非替尼的浓度

目的:建立一种简便、快速的高效液相色谱法测定人血浆中吉非替尼的浓度。方法:在100μL含待测药物的血浆中加入内标厄洛替尼和乙腈混匀沉淀蛋白。采用Agilent Eclipse XDB-C18柱(4.6 mm×150 mm,5μm),流动相为甲醇-10mmol.L-1醋酸铵溶液(70∶30),流速为1.0 mL.min-1,进样量为20μL,柱温为室温,分析时间为12 min。结果:吉非替尼在89.38～8938 ng.mL-1的浓度范围内线性关系良好(r>0.999),日内与日间精密度(RSD)<7.02%,准确度(RE)<±8.84%,所有稳定性考察结果均符合要求。结论:本文采用蛋白沉淀预处理方法以及高效液相色谱法测定人血浆中吉非替尼的浓度,能够满足临床试验中生物样品分析的需要。     

Phase matching of high harmonic generation in the soft and hard X-ray regions of the spectrum

We show how bright, tabletop, fully coherent hard X-ray beams can be generated through nonlinear upconversion of femtosecond laser light. By driving the high-order harmonic generation process using longer-wavelength midinfrared light, we show that, in theory, fully phase-matched frequency upconversion can extend into the hard X-ray region of the spectrum. We verify our scaling predictions experimentally by demonstrating phase matching in the soft X-ray region of the spectrum around 330 eV, using ultrafast driving laser pulses at 1.3-μm wavelength, in an extended, high-pressure, weakly ionized gas medium. We also show through calculations that scaling of the overall conversion efficiency is surprisingly favorable as the wavelength of the driving laser is increased, making tabletop, fully coherent, multi-keV X-ray sources feasible. The rapidly decreasing microscopic single-atom yield, predicted for harmonics driven by longer-wavelength lasers, is compensated macroscopically by an increased optimal pressure for phase matching and a rapidly decreasing reabsorption of the generated X-rays.