Simultaneous development of six LC-MS-MS methods for the determination of multiple analytes in human plasma

Traditional sequential single analyte method development is both time-consuming and labor-intensive. In this report, a concept of simultaneously developing multiple liquid chromatography coupled with tandem mass spectrometry (LC–MS–MS) methods were proposed. Mass spectrometric and chromatographic conditions as well as sample preparation methods for all analytes were optimized concurrently. Mass spectrometric conditions for six analytes, i.e. clonidine (CLO), albuterol (ALB), fentanyl (FEN), ritonavir (RIT), naltrexone (NAL), and loratadine (LOR), were established simultaneously using the Sciex Analyst software. LC–MS–MS sensitivities obtained using gradient elution methods on reversed-phase Inertsil ODS3 and normal phase Betasil silica columns were compared. Sample extraction methods using protein precipitation, liquid/liquid extraction, or solid-phase extraction (SPE) were evaluated. Recovery of analytes was determined. Matrix effects and interference due to endogenous compounds were investigated. Selection of a potential internal standard was discussed.     

FKBP12 is the only FK506 binding protein mediating T-cell inhibition by the immunosuppressant FK506

BACKGROUND: FK506-binding proteins (FKBP) are immunophilins that interact with the immunosuppressive drugs FK506 and rapamycin. Several FKBP family members such as FKBP12, FKBP12.6, and FKBP51 are expressed in T cells. It has been speculated that these FKBPs are possibly redundant in the immunosuppressant-induced T-cell inactivation. To determine the pharmacological relevance of multiple FKBP members in the immunosuppressant-induced T-cell inactivation, we have investigated the physiological responses of FKBP12-deficient and FKBP12.6-deficient mutant T cells to the immunosuppressive agent FK506. METHODS: FKBP12-deficient and FKBP12.6-deficient T cells were isolated from genetically engineered FKBP12-deficient and FKBP12.6-deficient mice, respectively. T-cell growth inhibitory assay was used to assess their responses to immunosuppressant FK506 treatments. RESULTS: We found that growth inhibition induced by FK506 is abolished in FKBP12-deficient cells but not in FKBP12.6-deficient cells. CONCLUSIONS: FKBP12 is the only FKBP family member that plays a key role in immunosuppressant-mediated immunosuppression.     

烧伤皮肤脂蛋白复合物诱导严重烧伤大鼠肺间质水肿的机理研究

目的：探讨烧伤皮肤脂蛋白复合物（LPC）对肺血管通透性的影响及其机理。方法：用30％Ⅲ度大鼠烧伤模型,提取外周血LPC;建立大鼠离体肺灌注技术;根据灌流液内容不同将实验动物分为4组：LPC组,烧伤血清组,正常血清组和对照组。测定灌流后5min和35min的肺重量增加（LWG）和液体滤过系数（Kf)及白蛋白通透性与表面积乘积（PS）。结果：（1）烧伤血清和LPC均能使LWG增加,以烧伤血清更为明显;（2）烧伤血清能使Kf和PS均增加,但以Kf的增加更为显著,LPC只能全程Kf增加而对PS无明显影响。烧伤血清对Kf的作用较LPC更为明显。结论：严重烧伤后,LPC具有诱发烧伤后肺水肿的作用,LPC可能介导或参与了烧伤后肺血管对小分子物质通透性的影响。     

桑白皮的薄层色谱鉴别方法研究

目的 研究桑白皮具有专属性的薄层色谱鉴别法。方法 用薄层色谱法,取桑白皮与混淆品种根皮的乙醇提取物,选择不同的展开剂展开,在紫外灯下检视荧光或三氯化铁显色,找出桑白皮特有斑点。结果 以CHCl3-MeOH(5:1)为展开剂,三氯化铁显色,桑白皮色谱中,在与混淆品种色谱相应位置上,显一个特有的紫色斑点。结论 特有斑点可作为桑白皮专属性的定性对照品,波谱分析鉴定为桑根酮C。     

Phosphorylated tyrosine-containing proteins in primary lung cancer correlates with proliferation and prognosis

To determine the usefulness of tyrosine phosphorylation in evaluating biological characteristics, we attempted to evaluate the relationship between the amount of phosphorylated tyrosine-containing proteins and clinicopathological factors, cell proliferation and outcome in non-small cell lung cancer. To evaluate phosphorylated tyrosine-containing proteins we used 96 surgically resected materials of non-small cell lung cancer and normal peripheral lung, while immunohistochemical evaluation was performed. Cell proliferating ability was evaluated using the labelling index of proliferating cell nuclear antigen-positive nuclear staining cells. There were statistically significant differences between the expression levels of phosphorylated tyrosine-containing proteins of normal and cancerous tissues (P<0.0001). Evaluations based on clinicopathological factors apart from histopathological differentiation, showed no statistically significant differences of phosphorylated tyrosine-containing proteins expression. However, phosphorylated tyrosine-containing proteins correlated with cell proliferation activity evaluated (P((Low, High))<0.0001; P((Low, Int)) <0.0001; P((Int, High))<0.0001). Furthermore, non-small cell lung cancer cases with high expression and intermediate expression of phosphorylated tyrosine-containing proteins had a significantly shorter disease-free postoperative survival than those with low expression of phosphorylated tyrosine-containing proteins using log-rank analysis (P((Low, Int)) <0.0028; P((Low, High))=0.0002). Furthermore, phosphorylated tyrosine-containing proteins expression level statistically contributed to disease-free survival in Cox's proportional hazard model. Therefore, phosphorylated tyrosine-containing proteins in non-small cell lung cancer tissues seem to reflect its biological malignancy, and this evaluation may be valuable for constructing the most appropriate therapeutic strategy.     

原发性脑干出血65例分析

目的：通过原发性脑干出血的临床与头颅CT分析，探讨脑干出血的出血量、出血部位与脑干出血症状、预后关系。方法：利用头颅CT检查对65例原发性脑干出血进行病因、症状和出血量、出血部位研究。结果：65例患者死亡51例，存活14例。出血量在1—18ml，出血横径5～35mm。结论：（1）血肿横径〉20mm或出血量〉5ml；（2）出血破入脑室；二者预后不良。出血量小于3ml者的轻型患者预后良好。     

支气管动脉介入栓塞术治疗7例重症支气管扩张伴大咯血

目的:探讨重症支气管扩张伴大咯血介入治疗的临床应用价值.方法:对7例重症支气管扩张伴大咯血患者采用支气管动脉造影和血管内栓塞术.结果:7例重症支气管扩张伴大咯血介入栓塞止血成功7例.结论:支气管动脉栓塞术在救治重症支气管扩张伴大咯血是替代保守治疗和外科手术治疗的另一种有效治疗方法.     

加味四逆散对肝纤维化大鼠血清透明质酸、层黏连蛋白、Ⅳ型胶原及肝脏超微结构的影响

目的：研究观察加味四逆散对实验性肝纤维化的预防和治疗效果，初步探索其防治机理。方法：采用人血清白蛋白（HSA）复制免疫性肝纤维化大鼠模型，设秋水仙碱为对照，观察加味四逆散对肝纤维大鼠血清透明质酸，层黏连蛋白，Ⅳ型胶原（放免法）及肝脏组织病理学和超同结构的影响。结果：肝纤维化大鼠在应用加味四逆散防治后，血清HA，LN，Ⅳ－C含量明显降低，肝脏组织病理损害明显减轻，胶原沉积减少，纤维增生显著改善，肝细胞坏死得以修复，与病理模型组比较有显著性差异（P＜0．05）。结论：加味四逆散能有效地改善肝脏微循环，保护肝细胞，恢复肝功能，抑制星状细胞活性，减少肝内胶原蛋白合成和沉积，促进胶原降解，具有肯定的抗大鼠肝纤维化作用。     

西洋参经验鉴别的体验与探讨

西洋参经验鉴别是判断 其优劣，区分真伪的有效而简便的方法。通过研究西洋参经验鉴别的文献，西洋参种植、采收、加工和干燥过程对其质量的影响，总结多年来西洋参鉴别的工作经验，对西洋参药材在形体、质量、表面纹理、横断切面及滋味气味等层面，提出一些西洋参经验鉴别的观点。     

再论按自身规律发展中医

以抽象概念演绎藏象、证候假说模型的唯象认知方式是学术的更高层次。由抽象思维推演的中医逻辑概念是驾驶实证技术的科学，具有广泛的临床指导意义而区别于西方医学，这是中医学的一大学术特色。然而，在现实中总有一种强行以西医辨病的实证技术方式来“改造中医”的说法和做法。有鉴于此，在1997年发表的《按自身规律发展中医》一文的基础上，再就上述观点作深入阐述，呼吁要确实注意中医不能丢。