用RP-HPLC指纹图谱控制复方丹参滴丸中低波长紫外吸收指纹成分

目的采用双定性双定量相似度作为评价指标,建立了复方丹参滴丸（CDDP）中低波长紫外吸收指纹成分的控制方法。方法采用RP-HPLC法以Century SIL C18BDS柱（200mm×4．6mm,5μm）;以色谱指纹图谱指数F为目标函数优化选择指纹图谱检测条件,确定流动相为水-乙腈低压梯度洗脱,紫外检测波长：203nm,柱温：（30．00±0．15）℃,进样量：10μL。以双定性双定量相似度法对10批CDDP进行质量评价。结果以人参皂苷Rg1为参照物峰,确定18个共有指纹峰,建立了CDDP低波长紫外吸收指纹成分的HPLC指纹图谱。评价出10批CDDP的双定性双定量相似度均合格,表明此10批样品的低波长紫外吸收指纹成分的数量、分布比例和含量特征是十分相似的,表现出很好的质量均一性。结论本试验证明双定性双定量相似度法可有效控制CDDP中低波长紫外吸收指纹成分。     

慢性乙型肝炎患者外周血IL-1β、mIL-2R、IL-10的检测及意义

目的 探讨细胞因子IL-1β、mIL-2R、IL-10与慢性乙型肝炎患者血清五项标志物的相关性.方法 应用化学发光免疫分析技术(CLIA)检测65例慢性乙型肝炎患者血清中IL-1β、mlL-2R、IL-10的水平;根据血清五项标志物检测结果,将65例慢性乙型肝炎患者分成两组,分别为HBeAg阳性组(33例,经检测HBV DNA全为阳性)、HBeAg阴性组(32例,经检测HBV DNA全为阴性),同时设正常对照组30例.结果 慢性乙型肝炎患者中HBeAg阳性组、HBeAg阴性组IL-1β、IL-10浓度明显高于正常对照组,经统计学分析差异有统计学意义(P<0.05);HBeAg阳性组、HBeAg阴性组mIL-2R浓度明显低于正常对照组,经统计学分析差异有统计学意义(P<0.05).结论 在慢性乙型肝炎发病过程中,mlL-2R等细胞因子水平降低,IL-1β、IL-10等细胞因子水平升高,可导致机体免疫功能紊乱,同时与机体乙型肝炎病毒感染后的慢性化密切相关.     

A catenary model to study transport and conjugation of baicalein, a bioactive flavonoid, in the Caco-2 cell monolayer: demonstration of substrate inhibition

The transport and metabolism of baicalein (Ba) was studied in vitro and in Caco-2 cells. Protein binding of Ba with Caco-2 lysate showed that Ba was bound to two classes of sites: a higher affinity, lower capacity site (K(A1) = 27.6 +/- 4.7 microM(-1), n(1) = 10.6 +/- 0.6 nmol/mg) and lower affinity, higher capacity site (K(A2) = 0.015 +/- 0.0013 microM(-1), n(2) = 413 +/- 21 nmol/mg). Incubation studies of Ba with Caco-2 lysate showed substrate inhibition of both glucuronidation and sulfation, with K(m) values of 0.14 +/- 0.034 and 0.015 +/- 0.0053 microM, and K(I) values of 6.75 +/- 1.70 and 0.37 +/- 0.16 microM, respectively. In the Caco-2 monolayer, Ba (8-47 microM) displayed good apparent permeabilities (P(app)) across the membrane; P(app) was found to be increased with elevated loading concentration in both the absorptive and secretory directions. However, the efflux ratio was less than unity, negating the involvement of apical efflux transporters. The concentration ratios of Ba sulfate (BS) and glucuronide (BG) decreased with increased loading Ba concentration, suggesting that BS and BG are apically excreted via transporters, likely breast cancer resistance protein and multidrug resistance-associated protein 2, respectively. Data fit to the catenary model, composed of basolateral, cellular, and apical compartments, showed a low cellular unbound fraction (0.0019 +/- 0.00018), a high passive diffusion clearance (0.012 +/- 0.00029 ml/min/mg), and substrate inhibition, with sulfation being more readily saturated and inhibited than glucuronidation, as evidenced by smaller K(m) value (0.35 +/- 0.078 versus 1.95 +/- 0.57 microM) and K(I) value (0.58 +/- 0.20 versus 7.90 +/- 1.10 microM); these patterns paralleled those observed in the lysate incubation studies. The results showed that the catenary model aptly predicts substrate inhibition kinetics and offers significant and mechanistic insight into the transport and atypical metabolism of drugs in the Caco-2 monolayer.     

Novel mutations of mitochondrial DNA associated with type 2 diabetes in Chinese Han population

Mitochondrial single nucleotide polymorphisms (mtSNPs) have been reported to associate with type-2 diabetes mellitus (T2DM), but mtSNPs appear to be considerably different among different populations and regions. To determine mtSNPs in Chinese Han patients with T2DM, the entire sequences of the mitochondrial genomes from 72 T2DM Chinese (59 +/- 4 years) and 50 age-matched healthy subjects (controls) in Chongqing region of Western China were directly sequenced and mtSNPs were analyzed. We found that M8, M9, D, G, R and A haplogroups exist in Chinese Han population and the frequency of haplogroup M9 was significantly higher in patients with T2DM than in the controls (p = 0.0006, OR 0.06 [95% CI 0.008-0.476]). MtSNPs T3394C in NADH dehydrogenase subunit 1 (ND1), G4491A in ND2, T16189C and T16519C were found with significantly higher frequency in patients with T2DM than in the controls (T16189C, p = 0.0045; T16519C, p < 0.0001; T3394C, p = 0.0015; G4491A, p = 0.0015). In contrast, the frequency of C5178A in ND2 and A10398G in ND3 was higher in the controls than in patients with T2DM (C5178A, p = 0.014; A10398G, p = 0.0011). Our results indicate that mtSNPs T3394C, G4491A, T16189C and T16519C show susceptible tendency to T2DM and mtSNPs C5178A and A10398G seem to be genetic factors for against T2DM. These mtSNPs determined in our study is useful and could be used for early diagnosis and prevention of T2DM in Chinese Han population.     

硫酸羟氯喹联合氟康唑治疗绝经期妇女口腔扁平苔藓临床疗效观察

目的观察硫酸羟氯喹联合氟康唑抗念珠菌治疗绝经期妇女口腔扁平苔藓（OLP）中的临床疗效。方法128例确诊为OLP的绝经期妇女随机分为两组,治疗组66例采用硫酸羟氯喹联合氟康唑治疗,对照组62例采用硫酸羟氯喹治疗。结果治疗组治疗6月后糜烂型OLP患者中有效率明显优于对照组（P〈0.01）,非糜烂型OLP疗效与对照组比较差异无显著性（P〉0.05）。结论硫酸羟氯喹联合氟康唑治疗能提高糜烂型OLP疗效,减少复发。     

踝关节骨挫伤X线与磁共振成像的诊断价值

目的探讨踝关节骨挫伤X线与磁共振成像(MRI)的诊断价值。方法对40例踝关节骨挫伤分别行X线和MRI检查,并观察其表现。结果40例患者X线可诊断7例明显骨折,MRI可诊断出全部40例骨挫伤及骨折,部分合并韧带损伤及关节积液。结论X线平片为踝关节外伤的最基本检查,但MRI能早期明确显示踝关节骨挫伤和部位,优于X线平片。     

布地奈德／福莫特罗治疗哮喘为期三个月的非干预性调研

目的 评价和调查布地奈德/福莫特罗常规临床治疗哮喘的疗效和使用情况.方法 开展多中心、开放的、非干预性调查研究,观察哮喘患者使用布地奈德/福莫特罗治疗12周后的哮喘控制情况,以哮喘控制调查问卷(ACQ)与改良的哮喘控制调查问卷(ACQ5)平均分值为主要疗效观察指标.结果 共入选受试者360例,其中成人哮喘228例,儿童哮喘132例.经12周治疗后,所有患者ACQ平均分值由治疗前的2.03(成人2.20,儿童1.74)下降为0.60(成人0.78,儿童0.29),ACQ5平均分值由治疗前2.47(成人2.24,儿童1.76)下降为0.47(成人0.62,儿童0.20),差异均有统计学意义(P<0.0001).结论 布地奈德/福莫特罗在常规临床应用时可以有效地改善哮喘,并达到哮喘的良好临床控制.     

Vaccination strategies to enhance local immunity and protection against Mycobacterium tuberculosis

To determine the immunogenicity and protective efficacy of the Mycobacterium tuberculosis 10 kD culture filtrate protein (CFP10), and to evaluate strategies that enhance local immunity, we used C57Bl/6 DR4 mice that were transgenic for human HLA DRB1*0401, because CFP10 contains epitopes for DRB1*0401 but not for C57Bl/6 mice. Intramuscular immunization with a DNA vaccine encoding CFP10 elicited production of IFN-γ by systemic CD4+ T cells, and one intravenous dose of the CFP10-based DNA vaccine coated with polyethylenimine (PEI) stimulated IFN-γ production by lung CD4+ cells and reduced the pulmonary bacillary burden. We conclude that CFP10 is a potential vaccine candidate and that coating vaccines with PEI enhances local protective immunity to tuberculosis.     

急性冠脉综合征患者血清hs-CRP和sCD40L临床特点

目的观察急性冠状动脉综合征（ACS）患者血清超敏C反应蛋白（hs-CRP）和可溶性CD40配体（sCIMOL）的变化及临床意义。方法采用夹心法酶联免疫吸附测定分析（ELISA）及微粒子增强透射免疫分析法分别测定对照组、稳定型心绞痛（SAP）组、ACS组的血清sCD40L和hs-CRP水平,并观察经皮冠状动脉介入（PCD术对其的影响。结果（1）ACS组血清hs-CRP及sCIMOL水平均高于SAP及对照组（P〈0．01）,SAP组与对照组无统计学差异（P〉0．05）;（2）相关分析显示血清sCD40L水平与hs-CRP呈正相关（r=0．787,P〈0．001）;（3）SAP和ACS组PCI术后血清hs-CRP、sCD40L水平均较术前显著升高（P〈0．01）。结论ACS组患者血清hs—CRP、sCD40L水平显著增高,提示冠心病的不稳定状态,可能有助于临床识别和预测ACS。     

Tea polyphenols exerts anti-hepatitis B virus effects in a stably HBV-transfected cell line

In this study, the anti-HBV effects of tea polyphenols （TP） were examined. After cells were exposed to TP for 3, 6, 9 days, amounts of HBsAg, HBeAg and HBV-DNA released into the supernatant of the cultured HepG2 2.2.15 cells were detected. TP, to some extent, inhibited the secretion of HBsAg and strongly suppressed the secretion of HBeAg in a dose-dependent （P〈0.01） and time-dependent manner, with 50% maximal inhibitory concentration （IC50） value being 7.34μg/mL on the 9th day, but the time-dependence was not significant （P=0.051）. Expression of HBV-DNA in the supernatant of the cell culture also was significantly decreased in a dose-dependent fashion （P〈0.01）. The ICS0 of TP in inhibiting HBV DNA was 2.54 pg/mL. It concluded that TP possessed potential anti-HBV effects and may be used as a treatment alternative for HBV infection.