人食管癌组织中热休克蛋白70和糖调节蛋白94的表达

为探讨人食管癌组织中热休克蛋白 70 (HSP70 )和糖调节蛋白 94( grp94)的表达及意义 ,采用SABC法检测人食管癌组织HSP70和grp94的表达 ,镜下观察HSP70和 grp94在癌组织及癌旁组织中的表达特点。结果 :92 .6%( 5 0 /5 4)食管癌组织中存在HSP70的表达 ,癌细胞存在胞质、胞核表达强度及部位的差异 ;而 grp94的表达阳性率为70 .4% ( 3 8/5 4) ,以胞质表达为主 ,而癌旁组织表达较少。提示 :人食管癌组织存在HSP70和 grp94的表达异质性 ,呈现了胞质、胞核的表达差异 ,HSP70和 grp94在人食管癌组织的表达特点为以后肿瘤抗原肽肿瘤疫苗的提取和实验奠定了基础。     

MS患者CD94/NKG2A-bright和CD94/NKG2A-dim NK细胞亚群对IFN-β的差异性反应

目的： 分析多发性硬化（MS）进展型患者不同表型NK细胞亚群对临床主要治疗方法的反应性差异.方法： 分离患者外周血中的NK细胞, 以流式细胞术根据表面抑制性受体CD94/NKG2A表达情况分为两个亚群CD94/NKG2A-bright和CD94/NKG2A-dim.分别加入IFN-β, 测定两个亚群表面CD94/NKG2A变化及细胞增殖, 同时检测两种亚群分泌IL-10和TGF-β情况.结果： CD94/NKG2A阳性表达的NK细胞占25.5%, 其中CD94/NKG2A-bright和CD94/NKG2A-dim分别占其中的23.6%和76.4%.加入IFN-β, CD94/NKG2A-bright组增殖率明显低于CD94/NKG2A-dim组, CD94/NKG2A表达峰度变化不大.CD94/NKG2A-dim组中CD94/NKG2A表达显著增加.两个亚群分泌的IL-10和TGF-β与未刺激组相比, 均有明显差异.CD94/NKG2A-bright和CD94/NKG2A-dim组间亦有明显差异.结论： IFN-β通过诱导NK细胞CD94/NKG2A表达在非特异免疫系统中抑制NK细胞; 同时刺激IL-10 和TGF-β分泌进一步发挥对免疫系统的抑制.CD94/NKG2A-bright和CD94/NKG2A-dim对IFN-β反应有差异性.     

Human CD8+ intraepithelial lymphocytes: a unique model to study the regulation of effector cytotoxic T lymphocytes in tissue

Summary:  The epithelium of the human small intestine contains a large population of intraepithelial cytolytic αβ T-cell receptor (TCR) CD8αβ T lymphocytes (IE-CTLs), whose main role is to sustain epithelial integrity by rapidly eliminating infected and damaged cells. In mouse, the recognition of inducible/modified self-molecules, i.e. non-classical major histocompatibility complex (MHC) class I molecules, is mediated by the TCR and natural killer receptors (NKRs) co-expressed on the cell surface of a non-conventional autoreactive CD8αααβTCR cell subset. In contrast, in humans, the recognition of non-classical MHC class I molecules induced by stress and inflammation on intestinal epithelial cells (IECs) is principally mediated by NKRs expressed on conventional CD8αβαβTCR cells. By sensing microenvironmental signals of inflammation and stress through NKRs, IE-CTLs fine tune their TCR activation threshold. Furthermore, IE-CTLs under particular conditions, involving interleukin-15 upregulation, acquire the capacity to kill distressed intestinal epithelial cells in an antigen non-specific manner. Adaptive IE-CTLs appear hence to have autoreactive properties and modulate their immune response based on innate signals, reflecting the fitness of the tissue.     

Lack of PTEN expression in endometrial intraepithelial neoplasia is correlated with cancer progression

We tested the hypothesis that PTEN inactivation may stratify cancer progression risk among putative endometrial hyperplasias, classified prognostically by means of the morphometric D score (DS). The DS, calculated from 3 morphometric variables measured in routine hematoxylin-eosin-stained endometrial biopsy slides, is the most sensitive and specific method of endometrial cancer risk prediction currently available. Clinical outcomes of 103 women with endometrial hyperplasia on biopsy were tallied according to the DS. Seven (7/103; 7%) patients with carcinoma during follow-up were all distributed within the high-risk prognostic group (ie, DS <1 = endometrial intraepithelial neoplasia [EIN]) (7/21; 33% progression). None of the 82 cases with a DS higher than 1 progressed. All cases that progressed were PTEN null, indicating that this genotype is capable of further stratifying cancer progression risk in hyperplasias irrespective of histological categorization. However, only 16% of the PTEN-null cases progressed. When PTEN expression pattern was combined with EIN, the prognostic power was greatly increased (specificity from 63% for PTEN and 85% for EIN to 93% when combined; positive predictive value from 16% and 33% to 50%). We conclude that loss of PTEN expression is the first biomarker in EIN that increases the accuracy of the prognostic DS to predict cancer progression risk. Unless endometrial hyperplasias are stratified by histological morphometric D-Score, PTEN has a low positive predictive value.     

Association of stress proteins with autoantigens: a possible mechanism for triggering autoimmunity?

Patterns of autoantibody production are diagnostic of many autoimmune disorders; the recent observation of additional autospecificities towards stress-induced proteins may also provide insight into the mechanisms by which such responses arise. Grp78 (also known as BiP) is a target of autoaggressive B and T cell responses in our murine model of anti-Ro (SS-A) autoimmunity and also in rheumatoid arthritis. In this report we demonstrate reciprocal intermolecular spreading occurs between Ro52 and Grp78 in immunized mice, reflecting physiological association of these molecules in vivo. Moreover, we provide direct biochemical evidence that Grp78 associates with the clinically relevant autoantigen, Ro52 (SS-A). Due to the discrete compartmentalization of Ro52 (nucleocytoplasmic) and Grp78 (endoplasmic reticulum; ER) we propose that association of these molecules occurs either in apoptotic cells, where they have been demonstrated indirectly to co-localize in discrete apoptotic bodies, or in B cells themselves where both Ro52 and Grp78 are known to bind to immunoglobulin heavy chains. Tagging of molecules by association with Grp78 may facilitate receptor mediated phagocytotsis of the complex; we show evidence that exogenous Grp78 can associate with cell surface receptors on a subpopulation of murine splenocytes. Given the likelihood that Grp78 will associate with viral glycoproteins in the ER it is possible that it may become a bystander target of the spreading antiviral immune response. Thus, we propose a model whereby immunity elicited towards Grp78 leads to the selection of responses towards the Ro polypeptides and the subsequent cascade of responses observed in human disease.     

The inhibitory NK cell receptor CD94/NKG2A and the activating receptor CD94/NKG2C bind the top of HLA-E through mostly shared but partly distinct sets of HLA-E residues

The human non-classical MHC class I molecule HLA-E is a ligand for both an inhibitory NK cell receptor (CD94/NKG2A) and an activating receptor (CD94/NKG2C). To identify HLA-E surface recognized by both receptors, especially to determine if both receptors recognize the same epitope, we made a series of individually Ala-substituted HLA-E proteins and analyzed their binding to CD94/NKG2A orCD94/NKG2C. Eight HLA-E mutations that significantly impaired HLA-E binding to CD94/NKG2A are all found in the top of alpha1/alpha2 domain of HLA-E. These results suggest that CD94/NKG2A binds a HLA-E surface equivalent to a NKG2D binding site on MICA. Of the eight mutations that impaired HLA-E binding to CD94/NKG2A, six significantly impaired HLA-E binding to CD94/NKG2C suggesting that CD94/NKG2C also binds a similar surface of HLA-E. Unexpectedly, the two HLA-E mutations (D69A and H155A) selectively abrogated HLA-E binding to CD94/NKG2A, not largely affected CD94/NKG2C. These results indicate that a mostly shared, but partly distinct set of HLA-E residues is discriminated by the two receptors.     

肩关节上关节囊重建治疗肩袖损伤适应证和移植物处理方法研究进展

目的对肩关节上关节囊重建治疗肩袖损伤适应证和移植物处理方法的研究进展进行综述。方法广泛查阅近年来国内外有关肩关节上关节囊重建治疗肩袖损伤的文献，并进行总结分析。结果肩关节上关节囊重建可以有效恢复肩关节上方稳定性，其手术适应证包括不可修复巨大肩袖撕裂、巨大肩袖撕裂合并肩关节假性麻痹、肩袖分层撕裂以及中或大的肩袖撕裂合并肩袖严重退变。为了获得较好的腱骨愈合以及降低移植物远期再撕裂率，必须选择合适厚度的移植物，并以适当的强度固定移植物并恢复其连续性。结论肩关节上关节囊重建的手术应用已发展至上关节囊加强重建，手术适应证从肩袖实质性巨大缺损扩展至肩袖严重退变。移植物处理是上关节囊重建手术成功的关键。     

锌指蛋白 A20 对兔腰椎间盘退变影响的实验研究

目的探讨锌指蛋白 A20 对兔腰椎间盘退变的影响。方法取 3 月龄新西兰大白兔 26 只，体质量 2.0～2.5 kg，经腹细针穿刺法制备 L_3、4、L_4、5、L_5、6 椎间盘退变模型，其中 24 只术后 4 周 MRI 检查明确造模成功，随机分为 4 组（n=6），于目标椎间盘中分别注射锌指蛋白 A20 过表达腺病毒（过表达 A20 组）、空载体腺病毒（空载体组）、PBS 液（对照组）、锌指蛋白 A20干扰腺病毒（干扰 A20 组）。于注射前 1 d 及注射后 1、2、3、6 d 行生物反应综合评分；注射后 2、4、8 周，各组行 MRI 检查并测量 T2 弛豫时间（T2 信号值）后，取材行阿利辛蓝染色观察椎间盘髓核细胞退变情况，免疫组织化学染色检测锌指蛋白 A20 以及椎间盘退变相关指标（Ⅱ型胶原、蛋白聚糖）的表达，Western blot 检测锌指蛋白 A20、NF-κB 结合蛋白［P65、磷酸化 P65（phosphate P65，P-P65）、Ⅱ型胶原、蛋白聚糖］、自噬相关蛋白［LC3 （LC3Ⅱ/LC3Ⅰ）、P62］以及炎症因子（TNF-α、IL-1β）的表达。 结果各组注射后各时间点生物反应综合评分均明显低于注射前 1 d（P<0.05）；注射后 6 d 干扰 A20 组评分明显低于其他组（P<0.05），其他组间比较差异均无统计学意义（P>0.05）。MRI 检测提示，注射后 2、4、8 周过表达 A20 组 T2 信号值均最高（P<0.05），2、4 周时干扰 A20 组最低（P<0.05），其余组间差异均无统计学意义（P>0.05）。阿利辛蓝染色显示，注射后 4 周过表达 A20 组蛋白聚糖含量最高（P<0.05）、干扰 A20 组最低（P<0.05）；8 周时过表达 A20 组蛋白聚糖含量显著高于其他组（P<0.05），其他组间比较差异无统计学意义（P>0.05）。免疫组织化学染色示，锌指蛋白 A20、Ⅱ型胶原、蛋白聚糖表达过表达 A20 组最高（P<0.05），干扰 A20 组上述蛋白表达最低（P<0.05）。Western blot 检测示锌指蛋白 A20、蛋白聚糖、Ⅱ型胶原、LC3 （LC3Ⅱ/LC3Ⅰ）蛋白相对表达量过表达 A20 组最高、干扰 A20 组最低，而 P-P65、TNF-α、IL-1β、P62 蛋白相对表达量过表达 A20 组最低、干扰 A20 组最高，与其他组比较差异均有统计学意义（P<0.05）；各组 P65 蛋白相对表达量差异均无统计学意义（P>0.05）。 结论锌指蛋白 A20 能通过抑制炎症反应，有效延缓兔腰椎间盘退变的进程。