耐辐射奇球菌PprI蛋白质在毕赤酵母菌中的高效表达及纯化

目的 利用真核毕赤酵母高效表达原核的耐辐射奇球菌pprI基因,建立高效表达及纯化PprI蛋白质的技术路线。方法 根据毕赤酵母密码子的偏爱性,改造耐辐射奇球菌pprI基因的编码序列,利用PCR技术全合成改造过的pprI基因,并在其N末端添加一个6×His标签。PCR产物经纯化后克隆到毕赤酵母表达载体pHBM-905A中,利用Cop I和Not I双酶切并回收线性化的目的片段后,转化毕赤酵母GS115菌株。将获得的毕赤酵母转化子诱导表达,SDS-PAGE、Western blot和质谱检测培养上清液,用Ni-NTA柱纯化目的蛋白,BCA法测定蛋白浓度。结果 新合成的耐辐射奇球菌pprI基因编码序列正确,毕赤酵母转化菌株的培养上清液经SDS-PAGE和Western blot均可检测到相对分子质量为43000的目的蛋白条带,经质谱检测证实该目的蛋白为耐辐射奇球菌PprI蛋白。选用Ni-NTA柱获得了大量纯化的PprI蛋白,应用浓度为250 mmol/L的咪唑淋洗时,PprI蛋白洗脱率最高。BCA法测得纯化蛋白浓度为0.35 mg/ml。结论 建立了一种新的适于毕赤酵母表达的耐辐射奇球菌pprI基因,成功构建了分泌表达PprI蛋白质的重组毕赤酵母工程菌株,建立了高效表达目的蛋白的技术路线,并获得了高效表达和纯化的PprI蛋白。     

基于文献计量学的耐辐射异常球菌研究态势分析

目的:分析国内外关于耐辐射异常球菌（DR）的研究现状,明确前沿与热点问题,初步评析其发展前景,为进一步的研究提供信息和方向。方法:以Web of Science和中国知网数据库为文献来源,检索2009至2018年主题词为"耐辐射异常球菌"、"耐辐射奇球菌"、" Deinococcus radiodurans...     

Comparison of T1 and T2 weighted images of the lumbar spine

In order to develop an optimal routine magnetic resonance (MR) spine scanning protocol, we have compared the relative efficacy of performing T1- and T2-weighted images in patients with various disorders of the lumbar region. Forty cases were randomly selected from studies performed from 1984 to 1987 and the TI- and T2-weighted images of each case were separated and interpreted blindly and independently by two neuroradiologists.Our results indicate no significant difference between T1- and T2-weighted images in the depiction of disc protrusion. The T2-weighted images were superior in depicting disc dessication, but the clinical significance of identifying a dessication disc remains uncertain. No significant difference in the depiction of osteophytes was seen between T1- and T2-weighted images. Cases of tethered cord, metastatic disease, and arachnoiditis were better delineated with T1-weighted images. The increase in signal of cerebrospinal fluid (CSF) with T2-weighting often obscured lesions within the spinal canal. In a signal case of postoperative diskitis, the T2-weighted images disclosed disc space abnormalities and epidural fluid collections not appreciated on T1 -weighted images. The T1-weighted images, however, did show thecal sac and adjacent epidural extension more clearly. In cases of metastatic disease, increase in the signal of metastases with T2 weighting often rendered them isointense to surrounding medullary bone.Given the lack of superiority of T2-weighted images over T1-weighted images in evaluating intervertebral disc protrusions and the superiority of T1-weighted images in depicting nearly all other abnormalities observed in this series, we no longer acquire T2-weighted images of the lumbar spine on a routine basis. In selected cases, such as evaluation of infected diskitis, T2-weighted imaging may have additional value.     

NOMOS-Kommentar zum Strafgesetzbuch (NK/StGB). Herausgegeben von Urs Kindh?user, Ulfried Neumann und Hans-Ullrich Paeffgen.

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变态反应性接触性皮炎体内外模型制作探讨

 目的 建立变态反应性接触性皮炎(allergic contact dermatitis,ACD)体内外模型。方法 采用二硝基氟苯(2,4-dinitrofluorobenzene,DNFB)建立小鼠耳廓ACD模型,计算耳肿胀度、HE染色;离体培养ACD小鼠T淋巴细胞,MTT法分析细胞增殖活性,免疫荧光检测CD4的表达水平,双抗体夹心ELISA法检测细胞因子IL-2和IL-4的含量。结果 小鼠ACD模型耳可见明显肿胀、大量炎性反应细胞浸润;模型小鼠T淋巴细胞体外培养能维持一定时程的高反应性及CD4的高水平表达,IL-2的峰值显著高于IL-4峰值。结论 成功建立了简单、快速、可靠的体内外ACD模型,可为评估变应原对人类健康的风险及筛选抗ACD免疫调节新药积累实验依据。     

新型细胞凋亡分子探针99Tcm-CP3-peptide的制备及在肺腺癌细胞系体内外的实验研究

目的 制备含天冬氨酸-谷氨酸-缬氨酸-天冬氨酸（DEVD）核心的多肽细胞凋亡分子探针99Tcm-CP3-peptide,研究其在肺腺癌细胞系A549化疗后体内外的生物学分布及SPECT显像。 方法 采用双功能螯合法合成99Tcm-CP3-peptide,分别于标记后0、1、2、4、6 h行高效液相色谱（HPLC）法检测放射性化学纯度。对经紫杉醇化疗的肺腺癌A549细胞进行体外细胞结合实验,对照组未经化疗,测量细胞内外放射性比值（Cin/Cout）,并用流式细胞仪检测细胞凋亡率。经紫杉醇化疗的荷瘤裸鼠尾静脉注射99Tcm-CP3-peptide 3.7 MBq,分别于5、15、30、60、120、240 min测其生物学分布,并行SPECT静态显像,勾画感兴趣区（ROI）,计算肿瘤/对侧正常肌肉组织的比值（T/NT）。取显像后的肿瘤组织进行苏木精-伊红染色,观察肿瘤组织的形态学变化。两组比较采用t检验,相关性研究采用双变量相关分析。 结果 99Tcm-CP3-peptide在0、1、2、4和6 h的放射化学纯度均>97%,合成率为（64.5±5.2）%。体外细胞结合实验结果表明,经紫杉醇化疗后24h,肿瘤细胞Cin/Cout为10.27±2.02,对照组Cin/Cout为1.09±0.03,化疗组为对照组的7.3倍。对同状态的肿瘤细胞行流式细胞检测,化疗组细胞凋亡率为（75.62±2.57）%,对照组细胞凋亡率为（3.42±0.32）%,化疗组和对照组Cin/Cout与流式细胞仪检测的凋亡细胞百分比呈明显正相关（r=0.970,P<0.05）。荷瘤裸鼠体内分布实验结果显示,99Tcm-CP3-peptide在血液中清除速度较快,且主要经肾脏代谢;心脏、脾脏、肺等重要器官放射性摄取值均较低;但在肝脏中摄取稍高且代谢速度较慢;肿瘤组织摄取值在注射后1 h达到峰值[（4.26±1.03）%ID/g]。荷瘤小鼠给药后1 h即可获得清晰图像,勾画ROI显示:化疗组T/NT为3.83±0.11,显著高于对照组的1.57±0.09,两者间差异有统计学意义（t=16.19, P<0.05）。病理结果显示化疗组肿瘤组织见大量核固缩、核碎裂及凋亡细胞;对照组仅见少量核固缩、核碎裂及少量凋亡细胞。 结论 99Tcm-CP3-peptide是一种靶向caspase-3的显像剂,具有较好的生物学分布,可应用于动物模型肿瘤组织凋亡显像,在监测化疗后肿瘤细胞凋亡方面有一定的临床潜在价值。      

Gerhard Rompe, Arnold Erlenk?mper, Begutachtung der Haltungs- und Bewegungsorgane.

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电离辐射对肺癌A549细胞miR-21体内外表达的影响及意义

目的 研究miR-21在肺癌组织及血清中的表达与肺癌预后及放疗因素的关系,进而探讨电离辐射对人肺癌A549细胞体内外表达miR-21的影响。方法 收集肺癌患者病理组织及血清样本,检测不同病理类型肺癌组织及血清中的miR-21的表达水平,同时检测是否接受放疗的非小细胞肺癌患者血清中miR-21的表达水平,并进行生存分析;以2、4 Gy X射线照射体外培养的A549细胞,并以A549细胞制备裸鼠肺转移癌模型,分别检测A549细胞与裸鼠血清及肺中miR-21的表达水平。结果 肺癌组织中miR-21高表达占60.0%;肺癌血清中miR-21高表达占50.5%,腺癌与鳞癌检出率差异有统计学意义（χ²=5.766,P<0.05）;87例非小细胞肺癌中放疗患者miR-21检出率66.7%显著高于非放疗患者39.6%（χ²=6.321,P<0.05）;Kaplan-Meier法生存分析显示,miR-21高表达患者预后明显低于低表达患者,差异有统计学意义（P<0.05）;Cox回归模型分析显示,miR-21高表达、区域淋巴结转移及放疗均为影响患者预后的独立危险因素。2、4 Gy X射线照射A549细胞后不同时间点miR-21表达显著升高（t=-7.552~-1.206,P<0.05）,miR-21在裸鼠血清及肺组织中的表达水平显著升高（t=-47.845~-2.356,P<0.05）。结论 电离辐射可上调A549细胞体内外miR-21的表达水平,可能与增强A549细胞侵袭转移能力相关。     

Total splenic infarct due to Aspergillus and AIDS

Invasive aspergillosis is a rare but important cause of morbidity and mortality in patients with AIDS. In the majority of patients, invasive aspergillosis involves the lungs. Extrapulmonary involvement with total splenic infarction has not been previously described. We illustrate a patient who developed total splenic infarction and septic emboli to the brain from Aspergillus.     

Determination of k0-values for the reactions Zr (n, γ) Zr and Zr (n, γ) Zr–Nb by irradiation in highly thermalized neutron flux

Determination of k₀-factors for zirconium isotopes was performed by co-irradiation of Zr and Au–Al. Due to the highly thermalized irradiation position at FRM-II, interferences from epithermal neutrons were found largely decreased for ⁹⁶Zr (n, γ) ⁹⁷Zr–^97mNb, the reaction with the highest Q₀-value in all (n, γ) reactions. Results showed that, the ⁹⁵Zr k₀-values from this work were the same as the recommended ones. For ⁹⁷Zr–^97mNb 743.4 keV gamma-line, the new k₀-value was 2.8% higher compared to the recommended value, which is not a significant difference. These results are helpful in clarifying the suspicion about the Zr k₀-factors.