CD44+/CD24-细胞在乳腺癌组织及细胞系中的数量与分布

Objective To study the distribution and quantity of CD44VCD24- cells in breast cancer tissue and the cell lines,and as well as its correlation with the expression of various breast cancer markers and molecular subtyping of breast carcinoma.Methods The expression of CD44 / CD24,estrogen receptor,progesterone receptor,HER2,human estrogen-induced protein PS2,bcl-2 and nm23 in 60 cases of invasive ductal carcinoma of breast were studied by either single or double immunohistochemical staining.The co-expression of CD44 and CD24 in 3 breast cancer cell lines (MCF-7,MDA-MB-468,and MDA-MB- 231) was also examined.Results The quantity and distribution of CD44 + /CD24- cells varied greatly and no specific patterns were identified.The percentage of CD44 + /CD24- in breast cancer was 65%.The amount of CD44+/CD24- cells did not correlate with the age of patients,lymph node metastasis,tumor　 size,molecular subtypes and expression of various breast cancer markers in breast carcinoma.The proportion of CD44+/CD24- cells in MCF-7,MDA-MB-468,and MDA-MB-231 cell lines was < 1%,5% and > 80% ,respectively.Conclusions CD44+ /CD24- cells are demonstrated in certain breast cancer tissues and cell lines.However,there is no relationship obtained between the quantity or the distribution of these cells and the molecular subtyping or the clinicopathologic parameters in breast cancer.     

Treatment plan modification using voxel-based weighting factors/dose prescription

Under various clinical situations, it is desirable to modify the original treatment plan to better suit the clinical goals. In this work, a method to help physicians modify treatment plans based on their clinical preferences is proposed. The method uses a weighted quadratic dose objective function. The commonly used organ-/ROI-based weighting factors are expanded to a set of voxel-based weighting factors in order to obtain greater flexibility in treatment plan modification. Two different but equivalent modification schemes based on Rustem's quadratic programming algorithms--modification of a weighting matrix and modification of prescribed doses--are presented. Case studies demonstrated the effectiveness of the two methods with regard to their capability to fine-tune treatment plans.     

Comparison of the unlabeled and labeled pre-mRNA splicing assays in vitro

Pre-mRNA splicing is a fundamental process required for the expression of most metazoan genes. It is carried out by the spliceosome that catalyzes the removal of non-coding intron sequences to ligate exons into mature mRNA prior to transport and translation. The purpose of our study is to explore whether the in vitro unlabeled pre-mRNA splicing assay could be performed as an alternative method of splicing reaction other than the radiolabeled one. Two different splicing methods in vitro , P labeled and unlabeled pre-mRNA as the substrates in the reaction, were investigated. The radiolabeled products were visualized by autoradiography while the unlabeled products were observed by Ethidium Bromide (EB) staining. As a result, although there are more unspecific bands in the EB staining assay than 32P labeled one, the RNA products of in vitro splicing could be observed clearly. This suggests that the unlabeled pre-mRNA splicing assay can be an optional substitution for the isotope-labeled assay.     

交互信息在心脏状态和心率研究中的应用

交互信息是一种检测系统之间相依性的方法 ,它可以同时检测线性和非线性相关。本文介绍了交互信息的计算方法和性质 ,讨论了它在单个时间序列上的应用 ,并将它应用于心脏状态研究上。利用实验动物的数据 ,我们发现了心搏的运动既不是随机的也不是周期的 ;并且 ,不同心脏状态下的交互信息有很大的差别 ,浅麻 ,机控呼吸和开胸状态下交互信息都相对较小 ,心肌损伤后有明显增大 ,提示心搏的交互信息与心脏健康程度有很大关系     

重组人bFGF的原核表达及其高效价抗血清的制备

目的 以重组人碱性成纤维生长因子为免疫原，制备高效价抗hbFGF抗血清。方法通过PCR方法改造5’编码区的12个密码子，构建hbFGF’原核表达载体并在大肠杆菌(E．coli)中表达，以纯化的hbFGF、免疫新西兰兔，制备高效价抗血清，用于重组hbFGF、的免疫印迹分析。结果经过改造的hbFGF基因在E．coli中获得较高水平表达。从可溶性部分纯化得到纯度95％以上的重组hbFGF，以该重组蛋白免疫兔子，在二次加强后以间接ELISA检测抗血清效价可达1：512000。免疫印迹分析显示该抗血清与E．coli中表达的重组hbFGF、和标准hbFGF、均有特异性反应，但与某些细菌蛋白存在弱交叉反应，经E．coli菌体蛋白吸附的抗血清，与菌体蛋白的弱交叉反应消失。结论以纯化的重组hbFGF为免疫原制备了高效价的特异性抗血清，经菌体蛋白吸附可消除存在的交叉反应性。     

30例麻疹暴发疫情的调查分析

目的调查本地某乡镇一次麻疹疫情的流行强度,描述其三间分布,分析暴发原因及影响因素。方法现场流行病学调查,酶联免疫捕获法检测麻疹IgM抗体,作免疫空白与麻疹发病关联的病例对照研究。结果2006年5月～7月,该乡镇发生30例麻疹局部暴发,其中29例儿童麻疹,1例成人麻疹,5例麻疹IgM抗体阳性。发病地点主要为小学和和幼儿园,罹患率分别为7.94%、7.87%。免疫空白与发病关系研究表明,初种免疫空白OR=7.2,X2=6.84,复种免疫空白OR=11.0,X2=9.48;影响免疫空白形成的社会因素调查,未接受麻疹疫苗初种或复种儿童的父母双亲常年在外务工占82.4%(28/34),有麻疹疫苗接种史儿童的父母常年在外务工占35.3%(12/34),二者有统计学差异(X2=15.54)。结论免疫空白是麻疹暴发形成的重要因素,农村儿童的父母常年在外务工形成隔代抚养教育对计免工作有负面影响。     

音猬因子的功能受体斑片在培养神经干细胞中的表达

目的　观察在培养的神经干细胞内是否有发育调控分子———音猬因子 (sonichedgehog)功能受体———斑片 (patched)表达。　方法　神经干细胞克隆在体外培养传代后 ,用patched的特异性引物对培养的神经干细胞进行RT PCR分析 ,PCR产物经克隆测序后 ,用地高辛标记克隆的探针 ,对神经干细胞进行原位杂交分析。　结果　神经干细胞克隆内大量的细胞均可表达sonichedgehog的功能受体patched ,patched阳性细胞间未见明显差别 ,克隆边缘与中央的patched分布也未见明显差别。　结论　sonichedgehog信号传导路可能在神经干细胞的增殖与分化过程中起重要作用。     

SARS病毒S1蛋白重组C端片段免疫效果的实验研究

为获得纯化的重组SARS病毒S1蛋白C端 ,研究其刺激机体产生针对SARS病毒免疫应答的规律和机制 ,将编码SARS病毒S1蛋白C端 311个氨基酸残基的基因克隆 ,并在原核表达系统中表达 ,纯化获得了的重组蛋白。利用SARS患者恢复期的血清 ,对纯化的重组S1蛋白进行血清学分析。结果表明 ,本研究中克隆表达的重组蛋白序列与公布的SARS病毒S1蛋白C端的序列相同 ,其编码的重组蛋白相对分子质量约为 5 90 0 0Mr。SARS患者恢复期的血清均与重组蛋白反应 ,在5 9 0 0 0Mr处形成特异性的反应条带 ,而来自SARS流行前的正常人对照血清则不能与重组蛋白反应。在本研究中获得的重组蛋白可以为研究SARS病毒识别宿主细胞受体的过程及其机制提供条件。     

中晚期重型乙型肝炎血浆置换与内科治疗生存率的比较

观察血浆置换、内科治疗对中晚期重型乙型病毒性肝炎生存率的影响,探索中晚期重型肝炎的治疗方法。方法观察在内科治疗基础上给予血浆置换患者的肝功能、并发症及疾病的转归,与同期仅予内科综合治疗的患者相比较,对相应的临床资料进行统计学分析,从而了解两种治疗方法的疗效。结果血浆置换组36例,18例好转、18例死亡,内科治疗组32例,13例好转、19例死亡,两组之间患者生存情况无统计学差异;肝功能指标(ALT、AST、SB、ALB、TC、ChE和凝血酶原时间(PT)也无统计学差异。结论与内科治疗相比较,在其基础上给予血浆置换并不能提高中晚期重型病毒性肝炎生存率;中晚期重型肝炎患者的预后决定于其肝功能衰竭的程度。     

肝脏特异性表达载体Kbpala/Alb-ATP7B 的构建及表达

目的：构建小鼠白蛋白启动子引导下携带野 生及常见突变型Arg778Leu、His1069Gln的人ATP7B cDNA的肝脏特异 性表达载体Kbpala/Alb-ATP7B，并检测其表达情况。方法:定点突变法获取人群中常见的Arg778Leu及His1069Gln两种ATP7B突变体，定向克隆将小鼠白蛋白启动子Alb序列及野生和突变的ATP7B cDNA依次亚克隆至转基因载体Kbpala上，得到可在肝脏特异性表达的正常及突变型人 ATP7B的转基因载体Kbpala/Alb-ATP7B。将其 短暂转染BRL细胞及BHK细胞，通过Western blotting检测其蛋白表达情况。结果:经酶切鉴定及定点测序证实，转基因载体Kbpala/Alb-ATP7B 构建成功；Western blotting显示仅在肝脏细胞实现表达。结论:带 有人类ATP7B cDNA的野生及常见致病突变型的转基因载体Kbpala/Alb -ATP7B构建成功并在肝脏细胞特异性表达。