小鼠下颔下腺中肥大细胞异质性研究

取小鼠下颌下腺,用甲苯胺蓝染色、Alcian蓝一藏红染色及免疫组织化学ABC法显示肥大细胞的异质性.结果表明;肥大细胞主要分布于间质的小血管、小叶间导管及神经节周围.此外还发现有些肥大细胞沿腺实质表面排列,有些肥大细胞伸出突起与相邻的神经元或肥大细胞接触.肥大细胞呈降钙素基因相关肽免疫反应阳性,但仅为相邻切片甲苯胺蓝染色的14 %.提示下颌下腺中肥大细胞的存在可能与腺体分泌活动及血流调节有关.     

Comparison of immunohistochemical markers in the differential diagnosis of adrenocortical tumors: immunohistochemical analysis of adrenocortical tumors.

Most adrenocortical tumors (ACTs) can be diagnosed directly by a combination of morphologic features and clinical findings. However, sometimes it may be difficult to distinguish ACTs from other neoplasms such as pheochromocytomas and some metastatic tumors, particularly for small biopsy specimens because they may be morphologically similar. Expression of calretinin has recently been suggested as a valuable immunomarker for the differential diagnosis between ACTs and other tumors; however, its diagnostic value is still under debate. To determine the diagnostic value of calretinin in Chinese patients with adrenocortical and non-ACTs, we employed both polyclonal and monoclonal anticalretinin to characterize the expression of calretinin in adrenal tissues and compared its expression with that of inhibin alpha, Melan-A, cytokeratin, or CD99 by immunohistochemistry in tissue microarrays and standard tissue sections of 414 specimens. Our results revealed that calretinin was expressed by adrenocortical cells, but not by the other cells tested and the percentage of calretinin-positive ACTs reached 99% when stained with polyclonal antibodies, which was higher than that with monoclonal anticalretinin (91.3%), anti-Melan-A (90.3%), antiinhibin alpha (81.6%). In addition, our results also revealed that ACTs were stained by cytokeratin (AE1/AE3) with variable degrees (58.7%). Furthermore, unlike anti-Melan-A that stained all metastatic malignant melanoma, anticalretinin did not recognize other tested tumors. Therefore, immunohistologic staining with polyclonal anticalretinin is more sensitive than other antibodies tested for the diagnosis of ACTs. However, monoclonal anticalretinin appeared to be more specific. Importantly, our data suggested that the fried-egg-like staining pattern, but not the mere cytoplasmic staining, was characteristic of anticalretinin staining in adrenocortical tissues. Notably, a few anticalretinin negative-ACTs were stained by other immunomarkers that we tested. Thus, the combinational characterization of calretinin (either by polyclonal or monoclonal antibody), inhibin alpha, and Melan-A expression is of great significance in the differential diagnosis of ACTs.     

In vitro degradation of silk fibroin

A significant need exists for long-term degradable biomaterials which can slowly and predictably transfer a load-bearing burden to developing biological tissue. In this study Bombyx mori silk fibroin yarns were incubated in 1mg/ml Protease XIV at 37 degrees C to create an in vitro model system of proteolytic degradation. Samples were harvested at designated time points up to 12 weeks and (1) prepared for scanning electron microscopy (SEM), (2) lyophilized and weighed, (3) mechanical properties determined using a servohydraulic Instron 8511, (4) dissolved and run on a SDS-PAGE gel, and (5) characterized with Fourier transform infrared spectroscopy. Control samples were incubated in phosphate-buffered saline. Fibroin was shown to proteolytically degrade with predictable rates of change in fibroin diameter, failure strength, cycles to failure, and mass. SEM indicated increasing fragmentation of individual fibroin filaments from protease-digested samples with time of exposure to the enzyme; particulate debris was present within 7 days of incubation. Gel electrophoresis indicated a decreasing amount of the silk 25 kDa light chain and a shift in the molecular weight of the heavy chain with increasing incubation time in protease. Results support that silk is a mechanically robust biomaterial with predictable long-term degradation characteristics.     

大鼠孤束迷走复合体儿茶酚胺能神经元向杏仁核的直接投射—HRP与免疫组织化学双标记研究

将HRP注入大鼠杏仁中央核(Ce)和基底外侧核(BL),发现在孤束迷走复合体(Sol/dmnx)尾段出现较多的逆行标记细胞。将WGA-HRP注入Sol/dmnx,在Ce和BL见到标记终末。用HRP逆行追踪和抗酪氨酸羟化酶(TH)抗体进行的免疫组化染色相结合的双重标记方法,发现从锥体交叉平面到最后区(AP)吻端平面,Sol/dmnx内有恒定的HRP逆行标记、TH样阳性和HRP/TH样双重标记的三种阳性神经元。双标记细胞主要分布于连合核(solC)、内侧亚核(solM),少量的位于间位核(solI)、背侧亚核(solD)及dmnx的背侧缘等。闩至最后区平面较多。阳性细胞皆以小型为主,多为圆形、三角形,soll中的则为长梭形。双重标记细胞占TH样免疫阳性细胞总数的18.8%,占HRP逆标细胞总数的90%,同侧为主,对侧偶见。本文对其功能意义作了探讨分析。     

4E-binding protein phosphorylation and eukaryotic initiation factor-4E release are required for airway smooth muscle hypertrophy

The molecular mechanisms of airway smooth muscle hypertrophy, a feature of severe asthma, are poorly understood. We previously established a conditionally immortalized human bronchial smooth muscle cell line with a temperature-sensitive SV40 large T antigen. Temperature shift and loss of large T cause G1-phase cell cycle arrest that is accompanied by increased airway smooth muscle cell size. In the present study, we hypothesized that phosphorylation of eukaryotic initiation factor-4E (eIF4E)-binding protein (4E-BP), which subsequently releases eIF4E and initiates cap-dependent mRNA translation, was required for airway smooth muscle hypertrophy. Treatment of cells with chemical inhibitors of PI 3-kinase and mammalian target of rapamycin blocked protein synthesis and cell growth while decreasing the phosphorylation of 4E-BP and increasing the binding of 4E-BP to eIF4E, consistent with the notion that 4E-BP1 phosphorylation and eIF4E function are required for hypertrophy. To test this directly, we infected cells with a retrovirus encoding a phosphorylation site mutant of 4E-BP1 (AA-4E-BP-1) that dominantly inhibits eIF4E. Upon temperature shift, cells infected with AA-4E-BP-1, but not empty vector, failed to undergo hypertrophic growth. We conclude that phosphorylation of 4E-BP, eIF4E release, and cap-dependent protein synthesis are required for hypertrophy of human airway smooth muscle cells.     

NOD小鼠胸腺细胞TCR介导的信号通路缺陷

目的 进一步研究NOD小鼠T细胞应答改变机理。方法 用抗TCR抗体、ConA激活NOD小鼠胸腺细胞，分析TCR介导的信号通路的水平。结果 与Balb／c小鼠胸腺细胞相比，抗TCR抗体诱导的增殖应答较弱，与年龄及NOD胸腺CD4^ CD8^-和CD4^-CD8^ SP细胞有关；rIL-2能部分恢复对TCR抗体应答的缺乏。NOD小鼠对PMA IONO和PMA anti—TCR-mAb应答正常，但对anti-TCRmAb IONO应答缺乏。结论 与年龄有关的NOD小鼠胸腺细胞对TCR抗体应答的缺乏与T细胞激活时上游PKC信号通路的缺乏有关。     

重组人免疫缺陷病毒Ⅰ型逆转录酶的纯化及其动力学性质

目的纯化重组人免疫缺陷病毒Ⅰ型逆转录酶（ＨＩＶ－１ＲＴ），筛选新的ＨＩＶ－１ＲＴ抑制剂。方法在适宜的培养条件下诱导工程菌Ｅ．ｃｏｌｉＪＭ１０９（ＰＫＲＴ２）可高效表达重组人免疫缺陷病毒Ⅰ型（ＨＩＶ－１）逆转录酶（ＲＴ）。应用ＤＥＡＥ－纤维素和磷酸纤维素离子交换柱层析法从细菌裂解液中分离、纯化重组ＲＴ。结果１升细菌培养液可得到１．１ｍｇ产物。ＳＤＳ－聚丙烯酰胺凝胶电泳分析显示所纯化的重组ＲＴ为由两个分子量分别为６６ｋＤ和５１ｋＤ的亚基组成的杂二聚体。酶活性测定结果表明，经纯化的重组ＲＴ具有很高的逆转录酶活性（比活力为１．４×１０４ｕｍｇ）。结论本文通过对ＲＴ反应条件的研究，优化了ＲＴ反应系统，并测定了磷甲酸钠（ＰＦＡ）对重组ＲＴ的抑制效应，结果表明ＰＦＡ对重组ＲＴ的抑制反应动力学机制与天然ＲＴ相同，从而进一步说明用此法纯化的重组ＲＴ可直接用于抗ＨＩＶ药物的筛选与评价。     

总责任护士主导的医护一体化护理模式在创伤骨科的应用

目的探讨总责任护士主导的医护一体化临床护理模式在创伤骨科的应用效果。 方法选择2016年7月至2017年6月在南京大学医学院附属鼓楼医院创伤骨科行手术治疗的骨折患者200例，按照入院先后顺序分为对照组90例(实施创伤骨科常规护理)和观察组110例(实施医护一体化工作模式，包括成立医护合作小组、实施总责任护士主导的医护一体化查房、个体化诊疗方案的共同决策、疼痛管理、早期出院计划与延续护理)。比较2组患者的术后护理效果和医护合作满意度。对数据行t检验和χ²检验。 结果总责任护士主导的医护一体化模式下，观察组患者术后72 h内数字评分法(NRS)评分为(4.09±1.64)分，疼痛干预有效率为77.57%，对照组术后72 h内NRS评分为(5.15±1.69)分，疼痛干预有效率为57.95%，2组比较差异均有统计学意义(P值均小于0.05)；观察组下肢深静脉血栓、泌尿系统感染、便秘的发生例数分别为2、1、3，对照组下肢深静脉血栓、泌尿系统感染、便秘的发生例数分别为9、7、11，两组比较差异均有统计学意义(χ²＝4.486、4.296、6.812，P＝0.027、0.036、0.009)。观察组术后康复开始时间为(21.48±6.41)h，术后住院时间为(11.08±2.19)d；对照组术后康复开始时间为(36.48±10.75)h，术后住院时间为(13.95±2.93)d，2组比较差异均有统计学意义(t＝11.670、－7.616，P值均小于0.05)。2组医护合作满意度结果显示，观察组患者信息的交流得分为(3.78±0.85)分，共同参与治疗或护理决策得分为(3.86±0.87)分，医师和护士的关系得分为(4.00±0.84)分，总分为(3.87±0.67)分；对照组患者信息的交流得分为(3.32±1.00)分，共同参与治疗或护理决策得分为(3.23±0.93)分，医师和护士的关系得分为(3.45±0.96)分，总分为(3.33±0.84)分，2组比较差异均有统计学意义(P值均小于0.05)。 结论总责任护士主导的医护一体化护理模式的应用，以亚专科为核心，重新整合医护资源，共同参与制订患者的诊疗护理方案，共同聚焦患者安全与质量，加强了医护间的配合，提高了医疗护理服务质量，提升医护合作满意度。     

层粘连蛋白受体、Ⅳ型胶原的表达及微血管密度与乳腺癌转移的关系

目的：探讨乳腺癌组织层粘连蛋白受体（LN－R）、Ⅳ型胶原（Col Ⅳ)、间质微血管密度与乳腺癌淋巴结转移的关系。方法：应用免疫组织化学S－P方法标记82例乳腺癌组织中LN－R、Col Ⅳ、FⅧRAg，并在第Ⅷ因子相关抗原（FⅧRAg)标记切片上计算微血管密度（MVD）。结果：LN－R表达强度及MVD的高低在乳腺癌有淋巴结转移组和无淋巴结转移组间有差异；ColⅣ在两组间无差异。结论：乳腺癌间质微血管密度增加及LN－R表达增强可作为预测肿瘤转移及判断预后的指标。