DF—88消毒洗涤剂杀菌抗病毒效果的初步观察

本文报道一种广谱、高效且有一定洗涤作用的杀菌抗病毒溶液——DF-88消毒洗涤剂，对其杀灭常见细菌和灭活乙肝病毒的效果及其稳定性、毒性作用等进行了初步观察。结果表明，当有效碘含量为2.54mg时，能完全杀死常见细菌；有效碘含量为23mg时，可有效地灭活乙肝病毒。此外，其LD_(50)为487mg/kg，皮肤刺激试验无任何充血、红斑等改变。     

乙型肝炎表面抗原“免疫逃避”突变体在哺乳动物细胞中的表达

利用乙型肝炎病毒ＤＮＡ开放框架上的ＢａｍＨＩ和ＨｐａＩ位点，酶切消化质粒载体ＰＥｃｏｂ６（含双拷贝ＨＢＶＤＮＡ），得到约９００ｂｐ的ＨＢＶ－Ｓ基因片断。将其插入到噬菌粒载体ＰＢｌｕｅｓｃｒｉｐｔｓｋ＋的ＳｍａＩ位点上。然后通过体外寡核苷酸介导的人工定点突变获得一系列（共１２种）Ｓ基因“免疫逃避”突变型。再通过ＥＢ病毒真核表达载体ｐＭＥＰ４上的ＢａｍＨＩ和Ｋｐｎｌ位点将噬菌粒ｐＢｌｕｅｓｃｒｉｐｓｋ＋上的Ｓ基因突变型片断定向克隆到ＰＭＥＰ４上，从而构建了含乙肝Ｓ基因突变型的重组质粒ｐＭＥＰ４ＨＢＳＭ。用其转染人肝癌传代细胞系ＨｅｐＧ２，经潮霉素选择，三周后获得抗性细胞克隆。经用抗ＨＢｓ单克隆抗体（含针对ＨＢｓＡｇ“ａ”抗原决定簇）检测除含变异体１４５（即１４５位上甘氨酸为精氨酸替代）外其余抗变异体ＨＢｓＡｇ均为阳性。经Ｗｅｓｔｅｒｎｂｌｏｔ证实变异体１４５，在分子量约为２３ＫＤ处有一特异ＨＢｓＡｇ蛋白带。     

Surgical strategies for treatment of giant or large intracranial aneurysms: our experience with 139 cases.

Giant or large intracranial aneurysms are the vascular neurosurgeon's greatest challenge. At our department, we have treated one hundred and thirty nine patients with giant or large intracranial aneurysms between 1975 and 2001. These included 37 partially thrombosed giant aneurysms. 75 aneurysms were giant (> 2.5 cm) and 64 were large aneurysms (2-2.5 cm). Three-dimensional computed tomography angiograms were performed in patients besides MRI angiography and digital subtraction angiography. These were found to be very valuable in the preoperative assessment of surgical anatomy of the aneurysm with respect to the branch arteries and perforators origin besides knowing the relations to the skull base. With our experience in surgical treatment of these 139 cases, we find that the basic technique is trapping and evacuation and not just clipping of the aneurysm neck but also reconstruction of the artery bearing the aneurysm, especially with wide-necked aneurysms. Use of multiple clipping, tandem clipping or dome clipping as per the intraoperative situation, is very helpful in dealing with giant aneurysms as also is the use of different types of clips like fenestrated clip with straight clip (combination clipping), booster clip, dome clips etc. While selecting surgical strategy for partially thrombosed giant aneurysm, securing the neck is most important. If the neck is too narrow to reconstruct, aneurysmectomy with anastomosis is one of the surgical strategies. An extracranial intracranial bypass should be considered in cases where clipping or parent artery ligation is expected to be associated with compromise of cerebral circulation.     

Polyclonal anti-PSA is more sensitive but less specific than monoclonal anti-PSA: Implications for diagnostic prostatic pathology

Prostate-specific antigen (PSA) production by nonprostatic tissues has been reported, casting doubts on its specificity. The immunohistochemical relative specificity and sensitivity of PSA expression using monoclonal and polyclonal anti-PSA was analyzed on 60 prostate carcinomas, 40 normal seminal vesicles, and 310 nonprostatic tumors. All nonprostatic tumors proved negative with both antibodies. However, 13 (32%) seminal vesicles showed immunoreactivity with polyclonal anti-PSA, but none showed immunoreactivity with the monoclonal antibody. The sensitivity of the 2 antibodies for prostate cancer varied with tumor grade. In Gleason pattern 3, both antibodies showed diffuse immunostaining in all cases. In Gleason pattern 5, polyclonal anti-PSA showed diffuse (>95%) tumor cell positivity in 18 cases (90%), while with the monoclonal antibody, 7 cases (35%) showed only focal (<10%) tumor cell immunoreactivity. Thus, monoclonal anti-PSA seems to be useful in small gland proliferations in which the differential diagnosis includes seminal vesicle, while for poorly differentiated neoplasms, polyclonal anti-PSA is considered superior. Sections of high-grade prostate cancer should be included as positive controls for PSA immunostaining.     

Benign tumors and tumor-like lesions of the adult kidney. Part I: Benign renal epithelial neoplasms

The spectrum of renal neoplasms has expanded in recent years. Although most of the work taking place in this field has concerned malignant neoplasms of the kidney, there have been significant improvements in our knowledge of benign renal tumors and tumor-like lesions, especially in renal cell adenoma, renal oncocytoma, and renal angiomyolipoma. Awareness and knowledge of these benign lesions is important because they are often included in the differential diagnoses of malignant tumors, with which they may be confused both clinically and pathologically. The authors review the topic of benign renal neoplasms and tumor-like lesions that occur in adults, emphasizing some of the newly described aspects of these lesions.     

急性鸭乙型肝炎病毒感染病毒清除机理研究

目的 :进一步阐明嗜肝病毒自然感染过程中病毒清除机理。方法 :7只 2～ 3月龄成年重庆麻鸭静脉接种 10～ 2 0mlDHBV阳性血清 (5× 10 8～ 1× 10 9genome) ,接种后每周采血检测外周血中DHBVDNA、DHBsAg和特异抗体的产生 ;感染后第10、35天分离外周血单个核细胞用于抗原特异细胞增殖实验 ;第 5、30、6 0天取肝组织标本进行DHBVDNASouthern杂交、DHB sAg免疫组化及肝组织病理检测。结果 :DHBV感染成年鸭在 1～ 2w潜伏期后出现急性、一过性感染 ,感染高峰期肝内存在多拷贝的DHBV所有复制中间体形式 ,包括cccDNA。进一步分析显示病毒血症的消失是在快速抗原特异细胞增殖反应和高滴度特异抗体产生之后 ;与此同时 ,整个急性感染期间 ,肝细胞并无明显的损害。结论 :非细胞直接损伤机制在嗜肝病毒清除过程中发挥了重要作用。     

RT-PCR-ELISA方法检测甲肝减毒活疫苗病毒滴度的初步研究

目的 应用核酸扩增产物测定的固相杂交酶联显色法(RT-PCR-ELISA)检测甲肝减毒活疫苗病毒滴度。方法 应用RT-PCR-ELISA。将标记有生物素的寡核苷酸引物所扩增的疫苗病毒基因产物。与微孔反应板上的特异性探针进行快速杂交，通过辣根过氧化物酶标记的链亲和素进行酶联显色。读取吸光度(A值)。判断结果。应用此法检测了11批甲肝活疫苗滴度。并与常规细胞培养法(CCID50)比较。结果 本方法与细胞培养法的敏感性相仿，具有简便、快速、特异的优点。结论 RTPCR-ELISA法有望代替常规细胞培养法应用于甲肝减毒活疫苗病毒滴度的检测。     

Chromophobe renal cell carcinoma: a comparative study of histological, immunohistochemical and ultrastructural features using high throughput tissue microarray

AIMS: In some cases distinction between chromophobe renal cell carcinoma (CRCC), oncocytoma and clear cell (conventional) renal cell carcinoma (eosinophilic variant) using routine light microscopy remains problematic. The present study investigates the level of agreement in the diagnosis of CRCC, as well as the histological features most frequently used for this diagnosis by two pathologists with a special interest in renal neoplasia. The sensitivity and specificity of immunohistochemical markers in cases with overlapping histological features in the diagnosis of CRCC were also studied. Electron microscopy was performed, as a diagnostic gold standard, on all of the cases. METHODS AND RESULTS: Thirty-two renal tumours with predominantly eosinophilic cytoplasm were reviewed in a blinded fashion by two pathologists. The diagnosis and morphological features used to render each diagnosis were tabulated. Validation of the utility of keratin 7 and 20, epithelial membrane antigen (EMA), vimentin, CD10, parvalbumin, RCC antigen, antimitochondrial antibody and Hale's colloidal iron was performed by the construction of a tissue microarray (TMA) master block. Based on histological criteria alone, overall agreement on the diagnosis of these tumours was reached in 69% of the cases, while there was total disagreement in 12%. In 59% of the cases, total agreement was reached in classifying the case as a CRCC based on histology alone. Kappa statistics for interobserver variability were calculated as only slight agreement (kappa = 0.3). The histological features most frequently associated with a diagnosis of CRCC were accentuated cell borders (87%) and a combination of hyperchromatic wrinkled nuclei (79%) and perinuclear halos (74%). The most sensitive and specific marker for CRCC was parvalbumin (sensitivity 0.91; specificity 1.0). The immunohistochemical profile of EMA+/ vimentin- was useful but had low specificity (sensitivity 0.75; specificity 0.4). CD10 had the highest sensitivity (1.0) but worst specificity (0.25) for CRCC. Keratin 7 had high sensitivity (0.83) but fairly low specificity (0.37) for CRCC. Hale's colloidal iron and the RCC antigen marker were not contributory. Finally, the antimitochondrial antibody was found to be fairly sensitive (0.83) for excluding CRCC. CONCLUSIONS: A small but significant proportion of renal tumours with cells having eosinophilic cytoplasm cannot be classified, even by experienced pathologists, based on histology alone. In these cases it is imperative to use markers with known sensitivity and specificity for the diagnosis of CRCC.     

人源腺病毒伴随病毒Ⅱ型单克隆抗体Fab段及其全抗体的研制

目的 运用噬菌体表面表达技术，获得基因工程抗腺病毒伴随病毒抗体Fab、IgG。方法 从酶联免疫吸附试验阳性的人外周血中分离淋巴细胞。提取总RNA，逆转录cDNA，聚合酶链反应扩增人IgG Fab轻、重链基因，利用pComb3载体构建噬菌体抗体库。用纯化的腺病毒伴随病毒为固相抗原筛选抗体Fab段，并在E.coli中进行分泌性表达。通过ELISA和间接免疫荧光试验、筛选和鉴定Fab抗体，并进行序列测定。然后将其重链和轻链基因克隆到全抗体表达载体pAC-L-Fc上。转染昆虫sf-9细胞，利用杆状病毒，昆虫细胞系统实现全抗体的分泌型表达，免疫沉淀试验鉴定它的抗蛋白。结果 分离到一株Fab克隆AAVs-31，腺病毒伴随病毒抗原和抗Fab抗体直接ELISA检测阳性，间接免疫荧光试验呈阳性，序列分析结果表明是一新的序列，所获得的基因为人源IgG Fab基因。由Gamma链和Kappa链组成。表达的全抗体ELISA反应、免疫荧光反应和间接免疫荧光试验均为阳性，免疫沉淀试验结果显示它结合病毒颗粒，不结合任一VP1、VP2、VP3单独衣鞘蛋白。结论 用噬菌体表面表达技术首次获得了抗腺病毒伴随病毒Ⅱ型单克隆抗体Fab段，并在真核系统中表达了其全抗体，它们识别的可能是衣鞘蛋白组装时形成的表位。     

超声处理对羊瘙痒病脑组织中PrPSc聚集状态的影响

目的 研究超声处理对感染羊瘙痒症仓鼠脑组织中PrP^Sc聚集状态的影响，寻找产生PrP^Sc低聚体的条件。方法 裂解液制备脑组织提取物，用各种超声条件处理不同阶段的脑组织提取物；以蛋白酶消化后的Western blot方法和图象分析系统观测PrP^Sc蛋白的分布和聚集状态。结果 适当的超声处理(15s共30次)可增加脑组织匀浆上清中PrP^Sc含量1．29～1．58倍；同样条件下超声处理可明显增加羊瘙痒因子263K感染仓鼠脑组织匀浆上清中PrP蛋白总量，而对正常对照仓鼠脑组织匀浆上清中PrP蛋白总量影响不大；对经常规高速离心获得的PrP^Sc的超声处理显示约90％的PrP^Sc存在于离心上清液中。结论 对感染动物脑组织进行超声处理可增加PrP^Sc的提取量，利于实验室检测。适当的超声处理可破碎大的相对分子质量的PrP^Sc聚集物，产生小的相对分子质量的PrP^Sc产物。