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.     

Degree of obesity is not associated with more than one intubation attempt: a large centre experience

Background

The role of obesity as a risk factor for difficult intubation remains controversial. We primarily assessed the association between body mass index (BMI) and difficult tracheal intubation.

Transfection of IL-2 and／or IL-12 genes into spleen in treatment of rat liver cancer

METHODS: A retroviral vector GCIL12EIL2PN encoding human IL-2 (hIL-2) and mouse IL-12 (raiL-12) fused gene and its packaging cell were constructed. The packaging cell lines contained of IL-2 and/or IL-12 genes were injected intrasplenically to transfect splenocyte at different time.The therapeutic effect, immune function and toxic effect were evaluated.RESULTS: The average survival times of the 4 groups using IL genes at days 1, 3, 5 and 7 after tumor implantation were 53.3&#177;3.7, 49.3&#177;4.2, 31.0&#177;2.1 and 24.3&#177;1.4 d respectively in IL-2/IL-12 fused gene group, 25.0&#177;2.5, 23.5&#177;2.0,18.3&#177;2.4 and 12.0&#177;1.8 d respectively in IL-2 gene treatment group, and 39.0+4.8, 32.0+3.9, 23.0+2.5 and 19.4+2.1 d respectively in IL-12 gene treatment group (P&lt;0.01, n=-10).In the IL-12/IL-2 fused gene treatment group, 30% of rats treated at days 1 and 3 survived more than 60 d and serum raiL-12 and hIL-2 levels were still high at day 3 after treatment. Compared with IL alone, NK cell activity was strongly stimulated by IL-2/IL-12 gene. Microscopy showed that livers were infiltrated by a number of lymphocytes.CONCLUSION: IL-2 and/or IL-12 genes injected directly into spleen increase serum IL-2 and IL-12 levels and enhance the NK cell activity, which may inhibit the liver tumor growth. The therapy of fused gene IL-2/IL-12 is of low toxicity and relatively high NK cell activity. Our data suggest that IL-2/IL-12 fused gene may be a safe and efficient gene therapy for liver tumor. The gene therapy should be administrated as early as possible.     

Patients with early diabetic heart disease demonstrate a normal myocardial response to dobutamine

OBJECTIVES: We sought to use quantitative markers of the regional left ventricular (LV) response to stress to infer whether diabetic cardiomyopathy is associated with ischemia. BACKGROUND: Diabetic cardiomyopathy has been identified in clinical and experimental studies, but its cause remains unclear. METHODS: We studied 41 diabetic patients with normal resting LV function and a normal dobutamine echo and 41 control subjects with a low probability of coronary disease. Peak myocardial systolic velocity (Sm) and early diastolic velocity (Em) in each segment were averaged, and mean Sm and Em were compared between diabetic patients and controls and among different stages of dobutamine stress. RESULTS: Both Sm and Em progressively increased from rest to peak dobutamine stress. In the diabetic group, Sm was significantly lower than in control subjects at baseline (4.2 +/- 0.9 cm/s vs. 4.7 +/- 0.9 cm/s, p = 0.012). However, Sm at a low dose (6.0 +/- 1.3), before peak (8.4 +/- 1.8), and at peak stress (8.9 +/- 1.8) in diabetic patients was not significantly different from that of controls (6.3 +/- 1.4, 8.9 +/- 1.6, and 9.6 +/- 2.1 cm/s, respectively). The Em (cm/s) in the diabetic group (rest: 4.2 +/- 1.2; low dose: 5.0 +/- 1.4; pre-peak: 5.3 +/- 1.1; peak: 5.9 +/- 1.5) was significantly lower than that of controls (rest: 5.8 +/- 1.5; low dose: 6.6 +/- 1.5; pre-peak: 6.9 +/- 1.3; peak: 7.3 +/- 1.7; all p < 0.001). However, the absolute and relative increases in Sm or Em from rest to peak stress were similar in diabetic and control groups. CONCLUSIONS: Subtle LV dysfunction is present in diabetic patients without overt cardiac disease. The normal response to stress suggests that ischemia due to small-vessel disease may not be important in early diabetic heart muscle disease.