构建以少突胶质细胞前体为主与人类早产儿脑室周围白质软化病理相似的动物模型

目的:建立以晚期少突胶质细胞前体为主、与人类早产儿脑室周围白质软化病理相似的可靠动物模型。方法:实验于2005-10/2006-06在上海交通大学医学院附属新华医院科研中心完成。2d龄和7d龄SD同窝清洁级新生大鼠各43只,雌雄不拘,以随机数字表法分为4组,即2d龄模型组(n=25)、2d龄假手术组(n=18)、7d龄模型组(n=25)和7d龄假手术组(n=18),脑室周围白质软化动物模型建立:结扎双侧颈总动脉,手术时间短于10～15min,术后将新生大鼠送入缺氧箱缺氧30min,混合气体为体积分数0.08的O2和0.92的N2,输入流量为1～2.5mL/min。假手术组游离双侧颈总动脉,但不予结扎和缺氧。将各组大鼠分别于术后1d测定脑梗死体积,术后2d进行少突胶质细胞系列免疫组织化学染色分析、脑片TTC染色观察脑梗死情况以及光镜下脑病理研究,术后21d进行电镜下病理研究。结果:实验86只新生大鼠,14只制作脑室周围白质软化模型死亡,余72只计入统计。①术后1d各组脑片大体观察:模型组脑内呈现大面积白色梗死区,多为大脑前、中动脉供血区域,2,7d龄模型组梗死体积分别为(53.45±33.90),(68.78±20.22)mm3,梗死百分比分别为(24.98±15.44)%,(11.84±4.14)%;假手术组脑片颜色鲜红,未见白色梗死区。②新生大鼠少突胶质细胞系列标志物免疫组织化学结果:2d龄新生大鼠脑白质内以少突胶质细胞前体为主,7d龄新生大鼠则以成熟少突胶质细胞为主。模型大鼠的相应少突胶质细胞系列阳性标记物的积分吸光度值均显著低于同日龄对照新生大鼠(P<0.05～0.01)。③各组大鼠术后2d光镜下脑病理检查结果:2d龄模型组新生大鼠的脑室周围以及皮层下白质呈现囊性坏死和细胞凋亡,而皮质神经元损伤轻微;而7d龄模型组在白质和皮质部位均呈明显损伤。④术后21d电镜下各组幼鼠脑病理检查结果:2d龄模型组幼鼠的脑白质内未见髓鞘形成,7d龄模型组幼鼠中见少量髓鞘形成,而同日龄假手术对照幼鼠的脑白质内则髓鞘形成正常。结论:通过对2d龄新生大鼠双侧颈总动脉结扎伴缺氧30min缺氧缺血法创建的脑室周围白质软化新生大鼠模型中存在少突胶质细胞前体和少突胶质细胞的明显受损和丢失,成功建立了2d龄新生大鼠以少突胶质细胞前体为主、与人类早产儿脑室周围白质软化病理相似的脑室周围白质软化动物模型。     

应用质谱、红外光谱及核磁共振解析验证合成物9,10-二羟基硬脂酸的纯化性

目的：为获得大量的9,10-二羟基硬脂酸（Dihydroxy Stearic Acid,DHSA）,应用质谱、红外光谱及核磁共振解析验证合成物为DHSA。 方法：实验于2007-01/05在解放军总医院营养科实验室完成。在烧瓶中加入油酸后,再加入甲酸、过氧化氢,并不断搅拌。在整个反应过程中维持反应温度在40℃,反应四五小时后,反复水洗油层并收集。在油层中加入3mol/L NaOH溶液25mL,于100℃水浴加热1h,趁热加入3mol/L HCl50mL,剧烈搅拌,析出白色沉淀。用水反复清洗使清洗液pH值为中性,收集沉淀物。在沉淀物中加入体积分数为0.95的乙醇10mL,加热溶解,抽滤,将滤液于0℃析出结晶,抽滤,将抽滤物用体积分数为0.95的乙醇10mL加热溶解,于0℃结晶,抽滤。同样方法再结晶2次,最后得到白色结晶物,晾干,收率68.8%。将结晶物分别上FTS-65A红外光谱仪、ISONS EA1108型元素分析仪,Varian INOVA 600型核磁共振仪,安捷伦XCR-trap液相质谱（LC-MS）联用仪进行分析和结构鉴定。 结果：①LC-MS分析提示合成的DHSA经过多次重结晶后,纯度较高,可达95%。MS分析提示DHSA相对分子质量为315.1,与理论值316.48相近。②DHSA元素分析实验值C68.78%,H11.53%,与理论值C68.77%,H11.58%接近。③红外光谱分析提示DHSA为含有两个羟基的十八碳长链脂肪酸。④核磁共振的氢谱和碳谱均证实合成的化合物为9,10-二羟基十八碳脂肪酸。 结论：合成的DHSA经过3次重结晶后,纯度可95%以上,质谱、红外光谱及核磁共振解析结果证实合成的化合物是9,10-二羟基硬脂酸。     

应用3.0T MRI评价缺血性脑血管病与鼻窦炎性病变的关系

目的:应用3.0TMRI从影像学角度探讨缺血性脑血管病与鼻旁窦炎性病变的关系及鼻旁窦炎性病变的MRI表现。方法:①分组及检测:2006-10/2007-08河北医科大学第二医院收治经临床及影像学证实为缺血性脑血管病患者218例为病例组,健康体检者218例为对照组,应用美国GE公司Signa Excite HD 3.0T高场强MR扫描仪常规行MRI及MRA检查。②评估指标:MRI图像观察以轴位T2-FSE、轴位T1-FLAIR序列为主,辅以轴位T2-FLAIR、矢状位T1-FLAIR序列,观察并记录脑实质MRI表现、鼻旁窦有无炎性病变。MRA图像观察以MIP重组后图像为主,辅以MRA源图像,观察并记录颅内血管异常情况。主要观察指标包括额窦、筛窦、蝶窦、上颌窦受累情况,各窦病变类型(黏膜增厚、炎性积液、黏膜下囊肿)及相应的MRI表现。结果:①病例组所有患者前循环及后循环系统血管均存在不同程度的动脉粥样硬化改变,143例患者脑实质出现缺血性病变;对照组脑实质及脑血管均未见异常改变。②病例组鼻旁窦有病变者123人次(56.4%),上颌窦病变96例,筛窦病变58例,蝶窦病变30例,额窦病变15例;对照组鼻旁窦有病变者54人次(24.8%),上颌窦病变8例,筛窦病变34例,蝶窦病变2例,额窦病变4例。病例组鼻旁窦炎性病变明显较对照组多(χ2=45.281,P=0.000)。相关系数C=0.91。结论:缺血性脑血管病患者鼻旁窦炎性病变发病率高,缺血性脑血管病与鼻旁窦炎性病变间有一定的相关性。     

聚乙二醇在猪组织工程瓣膜制备中的应用

目的:观察聚乙二醇法在组织工程瓣膜准备中的应用价值,比较聚乙二醇去细胞前后组织工程瓣膜的物理特性。方法:实验于2005-10/2006-03在华中科技大学同济医学院基础医学院生物化学系实验室完成。①实验分组:取猪10只,由于猪主动脉瓣为三叶瓣结构,共取得瓣叶组织30个,麻醉后宰杀取其心脏动脉瓣膜,分为去细胞组和对照组,每组各15个。②实验方法:去细胞组用聚乙二醇和DNase I处理;瓣叶组织放入1kg/L聚乙二醇,室温下浸泡30～45min,振荡器加以振荡;含抗生素磷酸盐缓冲液浸泡24h,反复3次洗脱;以5×104U/L DNase I液浸泡处理1h;对照组仅以含抗生素磷酸盐缓冲液浸泡24h,反复3次洗脱。③实验评估:苏木精-伊红染色、扫描电镜观察去细胞情况,吸光度(A)值,计算去细胞率(%)=(对照组A值-去细胞组A值)/对照组A值×100%。猪去细胞瓣膜条置于力学测试仪测定最大负荷、最大应力、最大应变和弹性模量。结果:纳入猪10只,均进入结果分析。①去细胞组织形态学观察:去细胞组猪瓣膜组织中看不到细胞成分,且细胞外基质结构保存完整,胶原纤维排列整齐,无明显断裂,仍呈波浪状平行排列,结构紧凑,弹性纤维结构清晰,组织无明显水肿。②DNA含量分析:聚乙二醇处理后去细胞百分率为95.32%。③生物力学检测:与对照组比较,去细胞组瓣膜组织最大负荷[(12.586±1.693),(10.242±1.435)N,P>0.05]、最大应力[(2.346±0.342),(1.877±0.572)N/mm,P>0.05]、弹性模量(15.152±1.579,14.549±0.678,P>0.05)、最大应变[(31.685±7.533),(28.118±6.045)mm/N,>0.05]等均无显著差异。P结论:聚乙二醇法去除细胞完全,细胞外基质保存完整,对组织机械性能无明显影响,适于构建组织工程瓣膜。     

The use of a recombinant immunoblot assay in the interpretation of anti- hepatitis C virus reactivity among prospectively followed patients, implicated donors, and random donors

Samples from prospectively followed recipients, their respective donors, and a cohort of random donors were used to evaluate the specificity and efficacy of a recombinant immunoblot assay (RIBA) as an adjunct to anti-hepatitis C virus (HCV) testing by enzyme immunoassay (EIA). RIBA reacted (RIBA+) in 100 percent of patients who developed hepatitis associated with anti-HCV seroconversion documented by EIA and in 100 percent of the EIA-positive (EIA+) donors implicated in these cases. In contrast, RIBA reacted in none of 10 recipients who were EIA+ but did not develop hepatitis, in none of 7 EIA+ patients with hepatitis B or cytomegalovirus infection, in 33 percent of EIA+ donors who were not implicated in hepatitis transmission, and in 37 percent of EIA+ random donors. Hence, the vast majority of EIA+ individuals who have ancillary evidence of HCV infection react on RIBA, whereas the majority of EIA+ individuals in low-risk settings do not react (RIBA-negative, or RIBA-). There was a strong association between RIBA reactivity and the presence of a surrogate marker (elevated alanine aminotransferase [ALT] and/or antibody to hepatitis B core antigen); 43 percent of RIBA+ implicated donors had a surrogate marker as compared to none of 14 EIA+, RIBA- donors. Among EIA+ random donors, 77 percent of those with a surrogate marker were RIBA+, as compared with 29 percent of those without a surrogate marker. In addition, in EIA+ donors, RIBA reactivity correlated with the extent of ALT elevation; 86 percent of those with an ALT greater than 135 IU per L were RIBA+ compared with 18 percent of those with an ALT less than 30 IU per L.(ABSTRACT TRUNCATED AT 250 WORDS)     

应用遥测肌电技术实测分析人体助行带的生物力学作用

目的:人体助行带为一种随身佩带的动力装置,应用遥测肌电技术测定和分析自行研制的人体助行带的生物力学作用机制。方法:实验于2005-12/2006-06在天津体育大学运动生物力学实验室进行。选取天津市天津医院30名受试对象,均对本实验知情同意。按照生物力学的测试要求,首先对受试者进行5次以上行走状态下的遥测肌电测定,然后在同等条件下对受试者佩带助行带实施对照研究。实验采用的助行带为自行研制的,由腰至足以双层弹力带作为动力装置的弹性结构。结果:行走摆动相大腿肌肌电:佩带助行带后肌内侧肌、肌二头肌、半腱肌肌电高于佩带前(P<0.01)。支撑相小腿肌肌电:佩带助行带后胫前肌、腓骨长肌、腓肠肌肌电高于佩带前(P<0.01)。结论:通过弹力带施加于下肢的外部载荷,能够起到增强下肢肌电以及产生助行的生物力学功效。     

Detection of antibodies to Trypanosoma cruzi among blood donors in the southwestern and western United States. II. Evaluation of a supplemental enzyme immunoassay and radioimmunoprecipitation assay for confirmation of seroreactivity

BACKGROUND: Chagas' disease, caused by the protozoan parasite Trypanosoma cruzi, is endemic to Latin America and may be transmitted in the United States via blood donated by infected immigrants. Blood- borne pathogens such as T. cruzi require supplemental testing for confirmation of seroreactivity. STUDY DESIGN AND METHODS: A study was undertaken to determine an optimal scheme for confirmation of seroreactivity in repeatedly reactive samples identified by the Chagas antibody enzyme immunoassay (EIA). The procedure for initial confirmation involves three purified antigens coated onto three separate polystyrene beads and uses an EIA format. If the sample is reactive with two of three or three of three antigens, it is confirmed as seroreactive. If none or one of three beads is reactive, the sample is indeterminate and subjected to a radioimmunoprecipitation assay (RIPA). The RIPA must demonstrate characteristic bands at 32, 34, and 90 kDa. RESULTS: When tested with sera from persons with potentially cross-reactive diseases (n = 39) or against a presumed negative population from southeast Wisconsin (n = 289), the confirmatory EIA had a specificity of 100 percent. Sensitivity was 100 percent (28/28) with xenodiagnosis-positive sera and 97.6 percent (80/82) with chagasic sera from Latin America. The RIPA showed a specificity of 100 percent in EIA- nonreactive samples (n = 100) and a sensitivity of 100 percent with both xenodiagnosis-positive (28/28) and chagasic (82/82) sera. CONCLUSION: The confirmatory EIA and the RIPA together provide a highly specific and sensitive means of confirming seroreactivity for antibodies to T. cruzi.     

Coinage metal complexes against breast cancer

Breast cancer is still the leading cause of cancer deaths among women worldwide, and new therapies to treat this dangerous disease are desperately needed. The serendipitously found anticancer drug cisplatin and its second-generation congener carboplatin appear to be promising drug systems for the treatment of breast tumors, in particular of multidrug resistant and highly aggressive triplenegative subtypes. In the wake of these platinum drugs, complexes of the coinage metals copper, silver, and gold were developed that showed enhanced selectivity for breast cancer while causing fewer and weaker side-effects. This review takes stock of the latest developments in the field of coinage metal anticancer drugs with an emphasis on their biological and mechanistic aspects. Pertinent literature is covered up to 2012.     

A novel SRY missense mutation affecting nuclear import in a 46,XY female patient with bilateral gonadoblastoma

Patients with disorders of sex development (DSD), especially those with gonadal dysgenesis and hypovirilization, are at risk of developing the so-called type II germ cell tumors (GCTs). Both carcinoma in situ and gonadoblastoma (GB) can be the precursor lesion, resulting in a seminomatous or non-seminomatous invasive cancer. SRY mutations residing in the HMG domain are found in 10–15% of 46,XY gonadal dysgenesis cases. This domain contains two nuclear localization signals (NLSs). In this study, we report a unique case of a phenotypical normal woman, diagnosed as a patient with 46,XY gonadal dysgenesis, with an NLS missense mutation, on the basis of the histological diagnosis of a unilateral GB. The normal role of SRY in gonadal development is the upregulation of SOX9 expression. The premalignant lesion of the initially removed gonad was positive for OCT3/4, TSPY and stem cell factor in germ cells, and for FOXL2 in the stromal component (ie, granulosa cells), but not for SOX9. On the basis of these findings, prophylactical gonadectomy of the other gonad was performed, also showing a GB lesion positive for both FOXL2 (ovary) and SOX9 (testis). The identified W70L mutation in the SRY gene resulted in a 50% reduction in the nuclear accumulation of the mutant protein compared with wild type. This likely explains the diminished SOX9 expression, and therefore the lack of proper Sertoli cell differentiation during development. This case shows the value of the proper diagnosis of human GCTs in identification of patients with DSD, which allows subsequent early diagnosis and prevention of the development of an invasive cancer, likely to be treated by chemotherapy at young age.     

Current concepts in stereotactic radiosurgery - a neurosurgical and radiooncological point of view

Stereotactic radiosurgery is related to the history of "radiotherapy" and "stereotactic neurosurgery". The concepts for neurosurgeons and radiooncologists have been changed during the last decade and have also transformed neurosurgery. The gamma knife and the stereotactically modified linear accelerator (LINAC) are radiosurgical equipments to treat predetermined intracranial targets through the intact skull without damaging the surrounding normal brain tissue. These technical developments allow a more precise intracranial lesion control and offer even more conformal dose plans for irregularly shaped lesions. Histological determination by stereotactic biopsy remains the basis for any otherwise undefined intracranial lesion. As a minimal approach, it allows functional preservation, low risk and high sensitivity. Long-term results have been published for various indications. The impact of radiosurgery is presented for the management of gliomas, metastases, brain stem lesions, benign tumours and vascular malformations and selected functional disorders such as trigeminal neuralgia. In AVM''s it can be performed as part of a multimodality strategy including resection or endovascular embolisation. Finally, the technological advances in radiation oncology as well as stereotactic neurosurgery have led to significant improvements in radiosurgical treatment opportunities. Novel indications are currently under investigation. The combination of both, the neurosurgical and the radiooncological expertise, will help to minimize the risk for the patient while achieving a greater treatment success.