Development and validation of a sensitive solid-phase extraction and high-performance liquid chromatography assay for the novel antitumour agent CT2584 in human plasma

A HPLC assay and solid-phase extraction technique from human plasma has been developed and validated for the novel anticancer agent CT2584, 1-(11-dodecylamino-10-hydroxyundecyl)-3,7-dimethylxanthine, which has recently completed a phase I trial at the Christie Hospital, Manchester under the auspices of the CRC phase I/II committee. Following addition of CT2576, 1-(11-octylamino-10-hydroxylundecyl)-3,7-dimethylxanthine, as internal standard, a solid-phase extraction cartridge (100 mg cyanopropyl) was used to isolate the drug CT2584 from human plasma. Analysis was performed by reversed-phase chromatography. CT2576 was used as internal standard at a concentration of 4 μg ml⁻¹ for the quantification of CT2584 from plasma for the duration of this work. The lower limit of quantification for the drug CT2584 in buffer using this assay was found to be 0.0122 μM (0.008 μg ml⁻¹) and 0.048 μM (0.027 μg ml⁻¹) when extracted from human plasma.     

花药培养对籼稻两用核不育系培矮64S的提纯与改良

以籼稻两用雄性核不育系（简称两用系）培矮６４Ｓ为供体,通过花药培养,获得了２６个愈伤系共１９６９株花粉植株。根据花粉一代（Ｈ１）的倍性和育性,可把它们分为三种类型,即ａ）单倍体：有１个愈伤系４１株全为单倍体植株;ｂ）不分离型：有１９个愈伤系的群体育性不分离,全部植株均在不育期表现败育,其它性状有差异但不显著;ｃ）分离型：其余６个愈伤系表现分离,其中３个各有２－７株单倍体植株出现,并且这些愈伤系中均     

脑膜炎球菌P64K蛋白作为新型抗原载体的研究进展

P64K是来自奈瑟氏属脑膜炎球菌的一种外层膜蛋白,发现于脑膜炎恢复期患者和接种过古巴VAMENGOCBC疫苗的个体血清内。编码P64K蛋白的基因称lpdA,其编码序列长1782bp。P64K蛋白含593个氨基酸,由两个结构域组成,有二氢硫辛酸酰胺酰基脱氢酶活性。P64K是抗脑膜炎疫苗的候选者。因为P64K的高分子量和强免疫原性,被认为是一种很有前途的抗原载体 。     

牛分枝杆菌mpb64基因的克隆、鉴定及其表达

以牛型分枝杆菌基因组DNA为模板,PCR方法扩增mpb64基因,纯化PCR产物并与pDM18-T载体连接、转化,经酶切及核苷酸序列鉴定为正确后,酶切产物亚克隆到原核表达载体pET30a(+)的KpnI／EcoRI位点,构建重组表达质粒pET30a+-mpb64,转化到大肠杆菌DE3内,以IPTG进行诱导,终浓度为1mmol／L,诱导产物进行SDS-PAGE电泳。结果表明,PCR方法成功扩增出mpb64基因,核苷酸序列测定验证了其正确性,重组表达质粒表达的pET30a+-mpb64融合蛋白相对分子量为30.4kDa,与实测相符。牛分枝杆菌pET30a+-mpb64的成功表达为牛结核病的诊断及新型疫苗的研究奠定了基础。     

Rare temporal bone pathology of the Singa calvaria from Sudan

Evidence has recently accumulated that the Singa calvaria from Sudan probably dates from Oxygen Isotope Stage 6 (>130 ka). Morphological studies have indicated a mixture of archaic and more modern human traits, but such analyses are complicated by the possibility that the vault is pathologically deformed, although the exact etiology has not been established. Now computed tomography (CT) has revealed that the right temporal bone lacks the structures of the bony labyrinth. The most likely cause of this rare pathological condition appears to be labyrinthine ossification, in which newly deposited bone obliterates the inner ear spaces following an infectious disease or occlusion of the labyrinthine blood supply. A possible cause of vascular compromise could have been the presence of an expanding acoustic neuroma in the internal acoustic meatus, which is suggested by a significantly wider right meatus compared with the left side. Interestingly, labyrinthine ossification is also consistent with the controversial diagnosis that an anemia caused the characteristic diploic widening at the parietal bosses, because prime etiological factors of ossification are among the common complications of some of these blood diseases. CT examination of the vault and a review of the literature suggest that a blood disorder may well have caused the unusual parietal morphology. Given the nature of these pathological conditions, the Singa individual must have experienced a period of considerable disability. The morphological evidence from the normal bony labyrinth on the left side and from the CT evaluation of the vault is consistent with the interpretation of Singa as a late archaic hominid or an early representative of Homo sapiens drawn from a population which might be directly ancestral to modern humans. Am J Phys Anthropol 107:41–50, 1998. © 1998 Wiley-Liss, Inc.     

赛加羚羊头骨的三维重建研究及其精确度评价

为建立赛加羚羊（Saiga tatarica）头骨三维可视化模型,进一步研究赛加羚羊头骨的解剖特征,建立赛加羚羊头骨的“数字化标本”,以提高其疾病诊治的准确率和物种保护的有效性。本研究以国家林业局甘肃濒危动物保护中心的赛加羚羊为研究对象,经锥形束CT扫描获得影像数据,运用MIMICS 20.0软件对赛加羚羊头骨扫描数据进行处理和三维重建,以建立赛加羚羊的“数字化头骨”用于形态研究。运用MIMICS 20.0软件对数字化头骨的26项形态学指标（n = 8）进行三维测量,且与头骨标本游标卡尺的测量值（n = 8）进行配对样本t检验分析,经与真实测量值比较后评价数字化模型及其三维测量的准确性。结果显示,26项形态测量指标均与真实头骨无显著性差异（P > 0.05）,例如颅全长游标卡尺测量值为（227.147 ± 10.646）mm,三维测量值为（227.130 ± 10.638）mm,P = 0.635,表明三维重建后的数字化头骨和真实的头骨标本高度相似,且“数字标本”能够实现任意角度的旋转、剖切和测量。本研究可为赛加羚羊头部疾病如骨折等的治疗及骨骼系统的三维可视化研究提供基础依据和技术支撑,同时可为CT扫描和医学图像三维可视化技术在野生动物的临床应用和相关研究提供理论基础。     

杆状病毒囊膜糖蛋白gp64基因启动子活性分析

GP64是杆状病毒在其发育循环第一时相所产生的芽生型病毒粒子（budded voirus,BV)的特异性囊膜糖蛋白。它在病毒入侵细胞过程中具有重要作用。为了探明杆状病毒gp64基因的表达调控。从所克隆的BmNPV和AcMNPVgp64基因ATG上游437-439bp启动子的序列分析发现。该启动子同时具有早期和晚期转录模体,将所构建的该启动子控制下荧光素酶报告基因（Luc)的非融合表达质粒分别转染Bm-N和Sf-21细胞进行瞬间表达分析,BmNPVgp64启动子能被允许宿主Bm-N细胞RNA聚合酶Ⅱ所识别,AcMNPVgp64启动子既能被允许宿主Sf-21又能被非允许宿主Bm-N细胞RNA聚合酶Ⅱ所识别,同时,这两个启动子的转录活性在各自的允许宿主细胞中被相应的病毒因子所反式激活2.4-4倍。将家吞核多角体病毒同源重复序列3（BmNPV homologous region-3,hr3)克隆到该启动子控制下的Luc报告基因下游,用所构建的质粒分别转染细胞,瞬间表达分析结果表达,BmNPVhr3能分别增强该启动子在Bm-N和Sf-21细胞中的转录活性13-22倍和7000-14000倍以上,同时,相应的病毒因子能反式激活插入了hr3的gp64启动子在各自允许宿主细胞中转录活性73-78倍,这暗示着BmNPVhr3除了具有病毒DNA复制原点和增强子的功能外,它在病毒的反式激活过程中起着重要的作用。     

Selective mutagenesis of lysyl residues leads to a stable and active form of delta 9 stearoyl-CoA desaturase

Stearoyl-CoA desaturase (SCD) is a short-lived integral membrane protein of the endoplasmic reticulum (ER) that catalyzes the insertion of a double bond in the delta 9 position of saturated fatty acids. Its expression has been difficult in heterologous systems. In this study, recombinant adenovirus vector was used to express both wild-type (wt) and engineered forms of rat SCD in human transformed kidney cells. In the engineered form of SCD, lysyl residues at positions 33, 35, and 36 were mutated to alanine (SCD K/A). The recombinant adenovirus also contains a cDNA encoding the green fluorescent protein (GFP). The stable reporter GFP was used to analyze the efficiency of transfection and the stability of expressed SCDs. The wt SCD was unstable upon expression, whereas expression of SCD K/A resulted in the stabilization of the protein. The proteasome inhibitor MG132 did not affect the rapid degradation of expressed wt SCD, implying that proteasome is not involved in this degradation. Functional analysis of microsomes from infected cells expressing SCD K/A resulted in the formation of holoenzyme with desaturase activity. Here we report engineering a stabilized form of a rapidly degraded membrane protein for production of an active mutant form of SCD. The adenovirus transformed cells may provide a model for the study of the effects of positive SCD expression.     

建立正常人肺血管容量的无创精确测量方法的初步报告*

目的: 由于传统呼吸调控环路忽略了对血液循环的决定性作用,肺（静脉）血管容量相关研究甚少,亟需建立肺血管容量测量方法。方法: 选择正常志愿者完成CT全肺扫描,图像数据经过计算机软件分析处理,从肺尖到肺底以40~50层进行肺野手工切划,相邻层间由计算机自动模拟连接,在去除干扰后进行全肺血管（≥0.6 mm）高精度三维立体成像技术处理,进而计算全肺和肺血管容积。结果: 12例正常志愿者从肺尖到肺底CT扫描图片层数为530±98（431~841）张。全肺和肺血管的总容积是3705±857（2398~5383）ml ,肺血管血液总的容积是125±32（94~201）ml。按肺静脉系统血管容量约为全肺血管血液容量一半计算,应该是63±16（47~100）ml。结论: 肺CT扫描数据分析三维立体成像建立肺血管容量无创测量方法精确可行。     

肺癌影像引导放疗人工图像配准方法分析

目的分析千伏锥形束CT（KVCBCT）引导肺癌放疗人工图像配准法的重复性。方法选择16例在我院行根治性放疗的非小细胞肺癌患者,每周行KVCBCT在线引导体位校正一次,获取患者KVCBCT影像。图像配准选择肺尖和椎体作为参考标记,在矢状位、冠状位和横断位等中心层面上配准患者KVCBCT影像和计划设计cT影像。比较同一名医生相隔一周两次配准,不同医生之间配准和医生与技术员之间配准结果的差异,用于评价KVCBCT引导肺癌放疗人工图像配准法的重复性。结果同一位医生相隔一周两次配准同一幅KVCBCT影像与计划设计cT影像,配准结果在患者左右（LR）、头脚（sI）和前后（AP）三个方向上,差值大于3mm所占的比例分别为：0,13％和6％。不同医生之间的配准结果在LR、SI和AP三个方向上,差值大于3mm所占的比例分别为：11％,19％和14％。医生与技术员的配准结果在LR、SI和AP三个方向上差值大于3mm所占的比例,分别为：16％,27％和27％。结论KVCBCT引导肺癌放疗人工图像配准法的重复性有待进一步提高。尤其表现为不同医生,医生与技术员之间应用该方法的重复性较差。KVCBCT引导肺癌放疗的图像配准方法需要进一步研究。