抗角蛋白自身抗体对豚鼠银屑病样皮炎的影响

目的 观察豚鼠抗角蛋白自身抗体（anti-keratin au-toantibody,AK auto Ab）滴度的消长对豚鼠银屑病样皮炎转归的作用。方法 应用心得安乳剂涂抹豚鼠耳廓造成银屑病样皮炎，采用腹腔注射纯化的豚鼠AK auto Ab（高滴度组）、皮下植入自身皮片消耗AK auto Ab（低滴度组）以及不作处理（正常滴度组）作为对照，根据组织病理变化评价豚鼠血清AK auto Ab滴度消长对皮炎转归的影响。结果 AK atuo Ab高滴度组银屑病样皮炎的恢复速度明显于对照组，而低滴度组皮炎恢复最缓慢。结论 AK auto Ab具有促进豚鼠银屑病样皮炎恢复的作用。     

In vitro keratin expression of hair cells.

Human hair follicles were isolated from the scalp by dispase and collagenase treatment and dispersed into a cell suspension by trypsin. These cells proliferated well and could be subcultured 7 to 8 times. The medium used was MCDB 153 HAA medium further supplemented with some amino acids, hydrocortisone, insulin, EGF, and bovine brain extract. The concentration of Ca++ was adjusted to 0.1 mM. Immunohistochemically, these cells were proved to possess keratins specific to hair forming cells.     

抗角蛋白自身抗体对白细胞介素1β诱发角朊细胞增殖的影响

目的：研究抗角蛋白自身抗体（ＡＫ ａｕｔｏ Ａｂ）对白细胞介素１β（ＩＬ－１β）诱发角朊细胞增殖的影响。方法：采用角朊细胞无血清培养方法，以＾３Ｈ－ＴｄＲ标示角朊细胞增殖程度，检测ＩＬ－１β刺激角朊细胞及ＡＫａｕｔｏ Ａｂ处理角朊细胞的增殖代谢。结果：１６００Ｕ／ｍｌ及以上浓度ＩＬ－１β对角朊细胞增殖作用有明显促进作用（Ｐ〈０．０１），正常血清浓度ＡＫ ａｕｔｏ Ａｂ对ＩＬ－１β引起的角朊细胞增殖     

Relationship between the expression of differentiation-specific keratins 1 and 10 and cell proliferation in epidermal tumors.

In normal epidermis, the expression of keratins 1 and 10 is associated with the loss of proliferative capacity and the onset of terminal differentiation. Keratins 1 (K1) and 10 (K10) are commonly expressed in the differentiating layer of benign tumors, but are lost during progression from the benign to the malignant state in skin carcinogenesis. Active gene constructs of mouse K1 and K10 were introduced into papilloma and carcinoma cell lines derived from keratinocytes to analyze the consequences of the expression of these keratins on the organization of the endogenous cytoskeletal network and on the mitotic activity of the recipient cells. Exogenous K1 integrated into the preexisting keratin K5/K14 network of both SLC-1 carcinoma and 308 papilloma cells. The formation of a recombinant cytoskeleton was more restricted for K10 than for K1 and appeared to be related to a requirement for cessation of cell division before K10 could integrate. The integration of exogenous K1 filaments into the endogenous keratin network was compatible with sustained proliferation of SLC-1 carcinoma cells in vitro. However, the exogenous gene was not expressed in tumor grafts in vivo. In contrast, stable K1 or K10 transfectants could not be selected in 308 cells, suggesting that benign tumor cells expressing suprabasal keratins cannot sustain proliferation.     

人发角蛋白材料植入修复受损骨骼肌后降解过程的观察

目的阐明人发角蛋白(HHK)材料植入体内后的降解过程。方法7只新西兰兔随机分为术后1、3、6周实验组和正常对照组。实验组进行骨骼肌切除后植入HHK材料,按期进行常规形态学和泛肽组化观察。HHK材料由3种降解速度的F、B、Z组分混合编制而成。结果光镜形态学观察显示材料植入后第1周出现HHK材料毛小皮脱落,HHK材料呈均质状,表面附着巨噬细胞和多核巨细胞;到第3周时可见降解成颗粒的材料被巨噬细胞和多核巨细胞吞噬。泛肽酶组化显示第1周时HHK材料及其周围的巨噬细胞、多核巨细胞呈阳性反应;第6周时材料进一步降解,同时伴有新生肌肉。电镜形态学观察显示毛发被降解成小的颗粒状。结论HHK材料的降解过程中,泛肽系统首先在细胞外将大体积的HHK材料降解成颗粒,随后细胞内泛肽系统通路和溶酶体通路分别对吞入的材料颗粒进行降解,且具有协同作用;同时肌卫星细胞被活化形成新生肌组织。     

从大容量噬菌体抗体库中筛选人源性抗角蛋白scFv

目的从大容量噬菌体抗体库中筛选人源性抗角蛋白单链抗体(scFv),测定其特异性结合活性并进行基因序列分析。方法以混合分子量人表皮角蛋白包被固相,对构建的大容量噬菌体抗体库进行筛选,经4轮“吸附洗脱扩增”后,挑取单克隆,用ELISA法测定特异性结合活性,并对阳性克隆抗体基因进行DNA指纹分析及序列同源性分析。结果经4轮筛选,获得9株可表达抗人角蛋白单链抗体的克隆,酶切鉴定正确后,经DNA指纹分析及序列同源性分析证明为不同的抗体基因。结论利用噬菌体抗体库技术获得了人源性抗角蛋白单链抗体,为临床应用研究提供了具有更广阔应用前景的人源小分子抗体。     

RF、CCP和AKA联合检测在类风湿关节炎诊断中的评价

目的 联合检测类风湿关节炎（RA）患者血清中的类风湿因子（RF）、抗环瓜氨酸肽抗体（CCP）和抗角蛋白抗体（AKA），探索其在RA诊断中的意义。方法 对69例RA患者、163例非RF的风湿免疫病患者血清同时进行测定了RF（速率散射比浊法）、CCP（ELISA法）和AKA（间接免疫荧光法），并对三者之间检测的一致性以及与其他临床指标的关系进行了分析。结果 RF、CCP和AKA对RA的敏感性分别为79.7％（55／69）、68.1％（47／69）和29.0％（20／69），特异性分别为75．5％、98．2％和100％。CCP的检测阳性率高于AKA（P＜0．01），与RF无差异（P=0．057），且与二者的检测具有一定的吻合度（k分别为0．271和0．483，P〈0．01）；CCP阳性组患者病程和RF表达量高于阴性组患者（P＜0，05），但患者年龄、ESR和CRP等无差异（P＞0．05）。结论 CCP抗体对RA具有较高的敏感性和高度的特异性，且对RA的病情如预后等可能有一定意义，值得临床推广应用。     

人发角蛋白植入大鼠损伤脊髓部位的电镜观察

目的:脊髓外伤后的继发性病理改变,是影响脊髓神经组织再生修复的重要因素。采用人发角蛋白(humanhairkeratin,HHK)植入脊髓损伤(spinalcordinjury,SCI)部位,以期达到减轻继发性损害,诱导和促进损伤脊髓组织的再生。方法:采用改制Ⅱ型纽约大学(NewYorkUniversity,NYU)装置,在建立大鼠脊髓损伤模型基础上,将经过特殊处理后能在体内降解的HHK植入大鼠损伤脊髓部位,对植入后1,4,12,26周的损伤脊髓组织进行电镜观察。结果:第1周为急性炎症时期,HHK周围结构紊乱,集聚大量的炎症细胞,灰质出现坏死;第4周时,炎症细胞减少,巨噬细胞吞噬髓鞘,胶质细胞增生;第12周时,多核巨噬细胞出现在HHK周围,HHK开始崩解,崩解物被多核巨细胞所吞噬;第26周时,神经轴突沿HHK间隙排列生长,灰质中神经元数量增加,HHK周边细胞有序生长。结论:植入的人发角蛋白具有诱导神经胶质细胞增生,阻止脊髓空洞的形成,从而减轻了脊髓损伤组织的继发性伤害的程度,改善了神经元再生的外环境,并可以桥接诱导神经轴突定向再生的作用。     

Keratin inclusions alter cytosolic protein localization in hepatocytes

Aim:  Mallory bodies have been observed in various liver diseases, however, the precise mechanism and significance of these structures have yet to be determined.
Methods:  Previously we reported on the redistribution of cytosolic proteins to keratin inclusions in mutant keratin 18-transfected cells. In this study, we treated green fluorescent protein-tagged wild-type keratin 18-transfected cells with several proteasome inhibitors and performed immunofluorescent analyses.
Results:  Proteasome inhibitors induced intracellular keratin inclusions, and desmoplakin, zonula occludens-1 and β-catenin were relocated to keratin inclusions, while theintegral membrane proteins were intact. The cytosolic proteins, 14-3-3 ζ protein and glucose-6-phosphate dehydrogenase were also relocated to inclusions. Moreover, E-cadherin, a basolateral membrane protein, was present on both the apical and basolateral domains in inclusion-containing cells.
Conclusion:  These data are identical to those in the mutant keratin 18 transfection study and suggest that keratin inclusions induced by different treatments affect localization of various cytosolic components, which may influence cellular functions performed by these proteins.     

Evolutionary aspects of the development of teeth and baleen in the bowhead whale

In utero, baleen whales initiate the development of several dozens of teeth in upper and lower jaws. These tooth germs reach the bell stage and are sometimes mineralized, but toward the end of prenatal life they are resorbed and no trace remains after birth. Around the time that the germs disappear, the keratinous baleen plates start to form in the upper jaw, and these form the food‐collecting mechanism. Baleen whale ancestors had two generations of teeth and never developed baleen, and the prenatal teeth of modern fetuses are usually interpreted as an evolutionary leftover. We investigated the development of teeth and baleen in bowhead whale fetuses using histological and immunohistochemical evidence. We found that upper and lower dentition initially follow similar developmental pathways. As development proceeds, upper and lower tooth germs diverge developmentally. Lower tooth germs differ along the length of the jaw, reminiscent of a heterodont dentition of cetacean ancestors, and lingual processes of the dental lamina represent initiation of tooth bud formation of replacement teeth. Upper tooth germs remain homodont and there is no evidence of a secondary dentition. After these germs disappear, the oral epithelium thickens to form the baleen plates, and the protein FGF‐4 displays a signaling pattern reminiscent of baleen plates. In laboratory mammals, FGF‐4 is not involved in the formation of hair or palatal rugae, but it is involved in tooth development. This leads us to propose that the signaling cascade that forms teeth in most mammals has been exapted to be involved in baleen plate ontogeny in mysticetes.