Biodegradable poly(ethylenimine) for plasmid DNA delivery

Poly(ethylenimine) (PEI) has been known as an efficient gene carrier with the highest cationic charge potential. High transfection efficiency of PEI, along with its cytotoxicity, strongly depends on the molecular weight. Synthesis of cationic copolymers derived from the low molecular weight of PEI and hydrophilic poly(ethylene glycol) (PEG), which are water soluble and degradable under physiological conditions, was investigated for plasmid delivery. Hydrophilic PEG is expected to reduce the toxicity of the copolymer, improve the poor solubility of the PEI and DNA complexes, and help to introduce degradable bonds by reaction with the primary amines in the PEI. Considering the dependence of transfection efficiency and cytotoxicity on the molecular weight of the PEI, high transfection efficiency is expected from an increased molecular weight of the copolymer and low cytotoxicity from the introduction of PEG and the degradation of the copolymer into low molecular weight PEIs. Reaction conditions were carefully controlled to produce water soluble copolymers. Results from a gel retardation assay and zetapotentiometer indicated that complete neutralization of the complexes was achieved at the charge ratios of copolymer/pSV-β-gal plasmid from 0.8 to 1.0 with the mean particle size of the polyplexes ranging from 129.8±0.9 to 151.8±3.4 nm. In vitro transfection efficiency of the synthesized copolymer increased up to three times higher than that of starting low molecular weight PEI, while the cell viability was maintained over 80%.     

Measurements of spontaneous ultraweak photon emission and delayed luminescence from human cancer tissues

OBJECTIVE: The aim of this study was to measure spontaneous photon emission (SPE) and delayed luminescence (DL) from various human cancer tissues. MATERIALS AND METHODS: A photomultiplier tube attached to a dark chamber was used for the detection of ultraweak photon emission from cancer tissues in the chamber. The samples were illuminated with a 150 W metal halide lamp for the measurement of delayed luminescence. Frozen tissues were provided by the hospitals and preserved in saline solution in a CO2 incubator for 1 hour before starting the measurement of spontaneous photon emission. We successively measured the afterglows from the samples after 30-second irradiation of the lamp. The samples were divided into two groups: tumor tissues and normal tissues around tumor tissues. We presented experimental data and interpreted their characteristic patterns of spontaneous photon emission and delayed luminescence. RESULTS: Mean values of spontaneous photon emissions from the normal tissues and the tumor tissues were measured with the standard errors of the mean as 625 +/- 419 counts/minute/cm2 (n = 6) and 982 +/- 513 counts/minute/cm2 (n = 14), respectively. Peak values of the intensity of delayed luminescence from normal and cancerous tissues were 63 +/- 20 counts/ms (n = 6) and 48 +/- 12 counts/ms (n = 14). CONCLUSIONS: The intensity of spontaneous photon emissions from cancer tissues were mostly discriminated from those of normal tissues, and their delayed luminescent properties were investigated.     

Characterization of a family of chimpanzee adenoviruses and development of molecular clones for gene transfer vectors

The high prevalence of preexisting immunity to the commonly used adenoviral vectors, as well as the requirement for readministration of vector for multiple therapeutic applications, necessitates the development of a panel of immunologically distinct adenoviral vectors against which neutralizing antibodies are rare in human populations. We have completely sequenced three chimpanzee-derived adenoviruses, Pan 5, Pan 6, and Pan 7, and have molecularly cloned E1-deleted vector genomes from each as bacterial plasmids. All the E1-deleted vectors were grown to high titer in HEK 293 cells. Neutralizing antibodies to the chimpanzee adenoviral vectors were not detected in serum samples from human subjects. In vitro cross-neutralization using rabbit antisera and in vivo readministration experiments in mice demonstrated that antibodies against Pan 5, Pan 7, or Pan 9 cross-neutralize one another but do not neutralize Pan 6. These results indicate that chimpanzee adenoviral vectors may be useful as vaccines or gene therapy vectors in human populations and should allow applications that require multiple vector administrations.     

高血压病患者肺静脉血流的超声多普勒研究

应用脉冲多普勒技术对照研究了５８例正常人和５６例收缩功能正常的高血压病患者二尖瓣和肺静脉的血流频谱参数。结果显示：高血压病组二尖瓣舒张期血流参数Ｅｐ、Ｅｉ、Ｅ／Ａ较正常组显著降低，ＡＦＦ显著升高；其肺静脉血流参数Ｓｐ、Ｓｉ、ＳＦ显著升高而Ｄｐ、Ｄｉ显著降低，即心室收缩期肺静脉向左房的回流速度及回流分数增加，舒张期的回流则减慢。说明高血压病患者在心功能代偿期，不仅存在左室舒张充盈的异常，同时也存在肺静脉的回流异常。     

血小板膜糖蛋白Ib／IX／V复合物表达变化：健康者与2型糖尿病患者的样本分析

背景：研究已证实血小板活化参与了糖尿病血管病变的发生发展。血小板膜糖蛋白Ib／IX／V复合物是血小板膜上主要的糖蛋白分子之一，是血管性血友病因子和凝血酶的受体，在血小板活化中起重要作用。 目的：观察2型糖尿病患者血小板膜糖蛋白Ib／IX／V复合物及其组分糖蛋白Ibd表达的变化。 设计：病例一对照实验。 单位：华中科技大学同济医学院附属协和医院中西医结合科。 对象：根据1999年世界卫生组织制定的糖尿病诊断标准选择2005—12／2007-01本院门诊就诊的外周血小板计数〉50×10^9L^-1的2型糖尿病患者51例，男28例，女23例，纳入对象在检查前两周内未服用影响血小板功能的药物。根据病情控制情况将患者分为控制良好组（n=25）和控制不良组（n=26）：根据是否合并血管病变分为合并血管病变组（n=27）和无血管病变组（n=24）。选择同期本院健康体检者23名为健康对照组，年龄、性别与病例组匹配。纳入对象或家属对检测项目和试验知情同意。试验经医院伦理委员会批准。 方法：患者均在清晨就诊时抽取空腹肘静脉血进行检测。健康体检者于来院体检时进行相同检测。①生化指标检测：由本院检验科完成空腹血糖、糖化血红蛋白和空腹面胰岛素的检测。②糖蛋白Ib／IX／V复合物及其组分糖蛋白Ibα的表达的检测：采集清晨空腹静脉血3mL，38g／L枸橼酸钠抗凝后10g／L多聚甲醛固定45min。取50μL固定后全血加至流式检测管，同时分别向不同管中加入20μL糖蛋白Ib／IX／V复合物单克隆抗体CD42a—b-c-d和PE标记的糖蛋白Ibα单克隆抗体CD42b，混匀后室温避光孵育30min，然后CD42a—b-c-d标记的样品中加入20uLFITC标记的羊抗鼠IgG混匀后室温避光孵育30min。于FACS420型流式细胞仪上检测样本平均荧光强度。③血小板聚集率测定：参照文献检测血小?     

人EGFR胞外段原核表达及免疫原性研究

目的：研究人EGFR胞外段在原核本质的表达，纯化条件以及对小鼠的免疫原性。方法：以既往构建包含人EGFR胞外段基因的表达质粒为基础，采用基因工程方法构建pET原核表达载体。在大肠杆菌中表达目的蛋白，表达蛋白在变性条件下，通过Ni^2 柱亲和层析纯化以包涵体形式存在的His-Tag融合蛋白，SDS－PAGE检测其纯度。去余His-Tag的纯化蛋白在梯度尿素中透析复性。以复性蛋白免疫小鼠获得抗血清，western blotting及流式细胞仪器检测特异抗体的存在。结论：通过限制酶切分析及测序鉴定表明获得了人EGFR胞外段基因的pET原核表达载体，SDS－PAGE分析表明在IPTG诱导下可以表达90Kd目的蛋白其纯度大于95％，所产生抗体可以识别复性蛋白及表达于A549细胞上的EGFR。提示在大肠杆菌中表达的人EGFR胞外段重组蛋白具有免疫原性，可以进一步用于EGFR免疫研究。     

Determining the main risk factors and high-risk groups of breast cancer using a predictive model for breast cancer risk assessment in South Korea

This study was aimed at developing a predictive model for assessing the breast cancer risk of Korean women under the assumption of differences in the risk factors between Westerners and Koreans. The cohort comprised 384 breast cancer patients and 2 control groups: one comprising 166 hospitalized patients and the other comprising 104 nurses and teachers. Two initial models were produced by comparing cases and the 2 control groups, and the final equations were established by selecting highly significant variables of the initial models to test the accuracy of the models in terms of disease probability and predictability. Both the initial models and the final disease-probability models were confirmed to exhibit high degrees of accuracy and predictability. Major risk factors determined by comparing the patients with hospitalized controls were a family history, menstrual regularity, total menstrual duration, age at first full-term pregnancy, and duration of breastfeeding. Major risk factors determined by comparing patients with nurse/teacher controls were age, education level, menstrual regularity, drinking status, and smoking status. The predictive model developed here shows that risk factors for breast cancer differ between Korean and Western subjects in the aspect of breastfeeding behavior. However, identifying the relationship between genetic susceptibility and breast cancer will require further studies with larger samples. In a model with nurse/teacher controls, drinking and higher education were found to be protective variables, whereas smoking was a risk factor. Hence the predictive model in this group could not be generalized to the Korean population; instead, breast cancer incidence needs to be compared among nurses and teachers in a nurse-and-teacher cohort.     

抑制miR-133b通过靶向FOXP3对PD大鼠调节性T淋巴细胞、炎性反应和神经元凋亡的影响

目的探究抑制微小RNA(microRNA,miR)-133b通过靶向叉头盒蛋白3(forkhead box protein 3,FOXP3)对帕金森病(Parkinson’s disease,PD)大鼠调节性T细胞(regulatory T cells,Treg)的影响。方法 32只PD模型大鼠随机分为PD组和PD+miR-133b antagomir组(n=16),健康大鼠16只作为对照组。尾静脉注射miR-133b antagomir(300μg)来抑制miR-133b的水平。检测和比较各组大鼠神经功能、黑质损伤、细胞凋亡、炎性反应、Treg细胞水平、miR-133b、FOXP3 mRNA和蛋白表达水平;通过双荧光素酶报告验证miR-133b和FOXP3的靶向关系。结果 PD组的逃避潜伏期、旋转速率、细胞凋亡情况、IL-6、miR-133b水平显著高于对照组,穿越次数、IL-10、FOXP3 mRNA和蛋白表达量显著低于对照组(P<0.05)。PD+miR-133b antagomir组的逃避潜伏期、旋转速率、细胞凋亡情况、IL-6、miR-133b水平显著低于PD组,穿越次...     

Hyperprolactinemia due to pituitary metastasis: A case report

BACKGROUNDPituitary metastasis is an uncommon manifestation of systemic malignant tumors. Moreover, hyperprolactinemia and overall hypopituitarism caused by metastatic spread leading to the initial symptoms are rare.CASE SUMMARYA 53-year-old male patient was admitted to our hospital with complaints of bilateral blurred vision, dizziness, polyuria, nocturia, severe fatigue and somnolence, decreased libido, and intermittent nausea and vomiting for more than 6 mo. During the last 7 d, the dizziness had worsened. Laboratory investigations revealed overall hypofunction of the pituitary gland, but the patient had an elevated serum prolactin level (703.35 mg/mL). Preoperative magnetic resonance imaging revealed a tumor in the sellar region, accompanied by intratumoral hemorrhage and calcification. Thus, transnasal subtotal resection of the lesion in the sellar region was performed. The histopathological and immunohistochemical examinations of the resected lesion revealed metastasis of lung adenocarcinoma to the pituitary gland. Oral hydrocortisone (30 mg/d) and levothyroxine (25 mg/d) were given both pre- and postoperatively. Post-operatively, the clinical symptoms were significantly improved. However, 4 mo following the surgery, the patient succumbed due to multiple organ failure.CONCLUSIONHyperprolactinemia is one of the markers of poor prognosis in patients with carcinoma that metastasizes to the pituitary gland. Exogenous hormone supplementation plays a positive role in relieving the symptoms of patients and improving quality of life.