两种检测方法在寻常型天疱疮诊断中的作用

目的 研究桥粒芯糖蛋白-酶联免疫吸附测定(desmoglein enzyme-linked immunosorbent assay,Dsg-ELISA)法检测寻常型天疱疮(pemphigus vulgaris,PV)抗体在PV诊断中的作用,以期为临床提供参考.方法 用Dsg-ELISA、猴食道为底物的间接免疫荧光法(...     

寻常型天疱疮患者血清抗桥粒芯蛋白3抗体水平及其影响因素分析

目的    研究寻常型天疱疮患者血清抗桥粒芯蛋白3（desmoglein 3,Dsg3）抗体水平及其影响因素。方法    选取2017年6月至2020年12月于南京市口腔医院口腔黏膜病科就诊的寻常型天疱疮患者30例,检查患者口腔病损部位,评估口腔损害严重程度,并采集患者血清样本。采用ELISA检测血清样本中与致病相关的抗Dsg3抗体水平,并分析血清抗Dsg3抗体水平的相关影响因素。结果    寻常型天疱疮患者血清中抗Dsg3抗体水平为（116.52 ± 7.35）U/mL。抗Dsg3抗体水平与寻常型天疱疮患者年龄、性别及口腔病损部位均无关（均P > 0.05）,与口腔损害严重程度有关（Z = -3.070,P = 0.002）。结论    血清抗Dsg3抗体水平能反映寻常型天疱疮患者口腔损害的严重程度,可用于监控病情和指导治疗方案的调整,提高诊疗水平。     

二期梅毒口腔临床表现及血清学检测结果分析

目的    分析二期梅毒患者血清学检测结果与其口腔临床表现的相关性,为临床诊疗提供参考依据。方法   选取2016年1月至2020年12月于南京大学医学院附属口腔医院·南京市口腔医院就诊的二期梅毒患者30例进行回顾性分析,研究患者口腔临床表现（包括病损数目和临床病损分级）与血清学检测结果［快速血浆反应素环状卡片试验（rapid plasma reactin ring card test,RPR）滴度］的相关性。结果    纳入研究的30例患者中,男14例,女16例;青年组（≤ 44岁）11例,中年组（45 ~ 59岁）10例,老年组（≥ 60岁）9例;各年龄组患者的性别分布差异无统计学意义（P > 0.05）。患者的口腔黏膜病损表现为黏膜斑、黏膜炎,好发部位依次为唇、舌、咽、腭和颊部。与低RPR滴度组（< 1∶128）相比,高RPR滴度组（≥ 1∶128）患者口腔黏膜病损数目较多、临床病损分级较高,差异均有统计学意义（均P < 0.05）。结论    伴有口腔黏膜病损的二期梅毒患者RPR滴度与口腔黏膜病损数目及临床病损分级密切相关。     

寻常型天疱疮诊断和治疗的研究进展

天疱疮是一种严重的累及皮肤和（或）粘膜的大疱性自身免疫性疾病,可危及生命。根据靶抗原和临床表现的不同可分为多种类型,其中以寻常型天疱疮最为常见,约占80％。在寻常型天疱疮中,75％～80％最先表现为口腔病损,患者往往最先求治于口腔科,因此口腔科医生在天疱疮的鉴别、诊断和治疗的过程中扮演重要角色。     

细胞角蛋白19和间隙连接蛋白43在4-亚基硝氧喹啉诱发大鼠舌黏膜癌变过程中表达的研究

目的 研究4-亚基硝氧喹啉（4NQO）诱发大鼠口腔黏膜癌变过程中细胞角蛋白19（CK19）和间隙连接蛋白43（Cx43）的表达,探讨口腔黏膜癌变过程中CK19与Cx43的相关性。方法 利用4NQO诱导SD大鼠的口腔黏膜癌变,运用免疫组织化学的方法检测CK19、Cx43在口腔黏膜癌变过程中各阶段的动态变化。结果 在大鼠正常舌黏膜组织中,CK19阳性染色的细胞散在分布于黏膜基底层;随着大鼠舌黏膜上皮异常增生程度的增加,CK19表达于黏膜基底上层;在口腔鳞状细胞癌（OSCC）组织中,CK19阳性染色细胞分布在黏膜各层。CK19在正常舌黏膜、轻度上皮异常增生、中度上皮异常增生、重度上皮异常增生、OSCC组织中阳性表达率分别为30.00%、50.00%、58.33%、80.00%、91.67%,差异有统计学意义（P<0.05）。在正常舌黏膜中Cx43蛋白主要表达于大鼠舌黏膜上皮细胞的细胞膜上,上皮的基底层、棘层和颗粒层呈阳性染色。随着大鼠舌黏膜上皮异常增生程度的增加,Cx43的表达明显下降。Cx43在正常舌黏膜、轻度上皮异常增生、中度上皮异常增生、重度上皮异常增生、OSCC组织中阳性表达率分别为100.00%、85.71%、66.67%、40.00%、33.33%,差异有统计学意义（P<0.05）。结论 在大鼠舌黏膜癌变过程中,CK19蛋白表达水平随病变程度加重显著升高,提示CK19与口腔上皮细胞的癌变有关;Cx43蛋白表达水平随病变程度加重显著下降,Cx43表达下降是口腔黏膜癌变的早期事件。CK19与Cx43蛋白表达呈负相关,CK19和Cx43的联合检测对OSCC的早期诊断有重要的作用,有利于提高OSCC早期诊断的灵敏度和特异性。     

免疫荧光在口腔粘膜天疱疮诊断中的应用

寻常性天疱疮属于自家免疫性疾患,口腔粘膜是最好发的部位,据Laskaris等统计,55—65%以上病例原发于口腔粘膜,且病变可持续几周、几月甚至几年而仅局限于口腔粘膜。Rosenberg等(1976)曾报导过一例病变局限在口腔粘膜达13年之久。自Beutnet和Jordon(1964)用间接免疫荧光实验(Indirect immunofluorescence,IIF)在患者血清中发现特异性抗体以     

p73在口腔黏膜白斑和口腔鳞癌中的表达

目的：观察p73在口腔黏膜白斑和口腔鳞癌中的表达,了解p73在口腔鳞癌发生过程中的变化规律。方法：白斑病理标本20例,口腔鳞状细胞癌病理标本31例,另取正常口腔黏膜10例,采用免疫组化检测方法观察p73的表达。应用SPSS11.0软件包对实验结果进行χ^2检验和单因素方差分析。结果：口腔白斑和口腔鳞癌病变组织中p73的表达率分别为65%和90.3%,均显著高于正常黏膜组（20%）,且随组织恶性程度增高呈明显上升趋势,正常黏膜上皮组、白斑组与鳞状细胞癌3组间比较,差异有统计学意义（P〈0.05）。结论：p73的表达与口腔鳞癌的癌变过程密切相关,可作为口腔白斑癌变的标志物。     

p63蛋白在鼠口腔黏膜干细胞中的时相表达和意义

目的：检测p63蛋白及其5种亚型在鼠口腔黏膜组织和口腔黏膜干细胞分化前后的表达特点,探讨p63作为口腔黏膜于细胞标记的可能。方法：以鼠抗人／鼠p63蛋白和兔抗人,鼠Oct－4蛋白为一抗,分别采用免疫细胞荧光和免疫组织化学技术检测p63蛋白在口腔黏膜上皮干细胞克隆、鼠上腭黏膜以及Oct－4在鼠腭黏膜中的表达情况;采用RT—PCR法检测口腔黏膜于细胞在培养5d和14d时,p63蛋白5种亚型的时相表达及变化特点。结果：免疫荧光显微镜观察,培养细胞胞核中发出p63蛋白的绿色荧光和DAPI蓝色荧光。免疫组化显示,p63蛋白主要表达在鼠腭黏膜基底层并部分表达于棘层,但Oct－4只表达于基底层。RT—PCR显示,细胞在培养5d时,表达p63蛋白的TA、△N、α和β亚型,不表达γ亚型;在培养14d时,表达p63蛋白的TA和γ亚型,不表达△N、α和β亚型。结论：p63蛋白可在口腔黏膜干细胞和次分化细胞中表达,单独标记尚不足以精确示踪口腔黏膜干细胞,但是其亚型△Np63α和（或）△Np63β可能是口腔黏膜干细胞的特异性标记。     

口腔黏膜癌变过程中中心体的扩增及其意义

目的　了解口腔黏膜癌变过程中中心体的状况,探讨中心体的异常在口腔鳞状细胞癌(OSCC)发生发展中的作用及其在口腔黏膜癌变过程中的意义。方法　选取正常口腔黏膜(12例)、轻度上皮异常增生(2例)、中度上皮异常增生(8例)、重度上皮异常增生(12例),口腔高分化鳞癌(10例)、中分化鳞癌(15例)、低分化鳞癌(7例)的石蜡包埋组织,采用间接免疫荧光双重染色(γ-微管蛋白单克隆抗体及细胞角蛋白多克隆抗体)显示上皮细胞中的中心体,分析其在口腔黏膜癌变过程中的变化趋势及在各组间的差异。结果　正常口腔黏膜上皮表现为大小数目正常的中心体,但在72·73%(16/22)的上皮异常增生及84·38%(27/32)OSCC组织中观察到部分上皮或肿瘤细胞中出现异常中心体,表现为中心体直径的增加或数目的增多。具有异常中心体的细胞数目随上皮异常增生程度的增加及肿瘤分化程度的降低而增加,二者存在明显的正相关关系(P<0·01)。结论　中心体的扩增是口腔黏膜癌变过程中的早期事件并与口腔癌发生发展进程有关,口腔黏膜癌变过程中的细胞形态学改变与中心体扩增之间可能存在直接机制上的联系。从调控中心体循环入手治疗口腔癌前病损及OSCC,有可能成为口腔癌预防和治疗的新途径。     

白念珠菌口腔黏膜感染小鼠模型的建立及评估

［摘要］ 目的 建立白念珠菌口腔黏膜感染ICR小鼠模型,动态观察口腔黏膜及相应的全身感染情况,并对变化情况进行初步分析。方法 将白念珠菌接种于免疫功能低下小鼠口腔,采用肉眼观察口腔舌背病损改变、口腔内白念珠菌菌量检测、组织病理学等方法评估口腔内黏膜感染情况,检测体重、载菌量（肾、肝、下降结肠内粪便）评估全身感染情况。观察1周,并记录数据。空白对照组为接种生理盐水的免疫功能低下组。结果 ①小鼠接种白念珠菌后第1天口腔白念珠菌菌量较低,随后迅速增长,3~7 d内趋于稳定,稳定于106~107CFU/mL。同时口腔舌背病损也出现类似趋势,第1天未见明显伪膜,3~7 d内可见舌背伪膜存在。②病理切片PAS染色显示接种后3~5 d内白念珠菌形成菌丝侵入并破坏上皮,到接种第7天时,黏膜上皮多见酵母细胞。③小鼠接种白念珠菌后体重逐渐下降。粪便载菌量逐渐上升,第5天达到最大,但肾、肝无白念珠菌感染。空白组未见白念珠菌感染。结论 通过在ICR小鼠口腔内接种白念珠菌可以建立白念珠菌口腔黏膜感染动物模型。接种后白念珠菌感染程度在1周内存在变化。动物实验应根据感染状况选择合适的研究时段。     

Relationship between the serum autoantibody titers and the clinical activity of pemphigus vulgaris

With human tonsillar epithelium as a substrate for the indirect immunofluorescence assay, a statistically significant relationship was demonstrated between the clinical activity of the disease and the autoantibody titers in seventy-eight serum samples obtained from patients with pemphigus vulgaris. High pemphigus antibody titers were associated with severe ulceration, intermediate titers with moderate lesions, and low titers with mild pemphigus. However, the sequential studies revealed that this relationship was not always consistent throughout the course of the disease, and in three out of fourteen patients the correlation was either poor or negative. These results indicate that serial pemphigus antibody titers may not be consistent enough to be used reliably as the sole guide for monitoring disease activity.     

Clinical and cytological features of oral pemphigus

Nine patients with pemphigus vulgaris and one with pemphigus vegetans were studied. All ten patients had oral lesions, five of them skin involvement as well. The patients were followed up for variable periods at different intervals. At each visit, their clinical condition was assessed as mild, moderate, severe or remissive. At examinations, oral light microscopical smears and blood samples for indirect immunofluorescence were taken. From one patient with pemphigus vulgaris, smears from skin blister fluid were also studied. The highest intercellular antibody titers were found in severely affected patients, and those at the mild stage of the disease usually gave low or negative titers. A correlation between the number of acantholytic cells in smears and the clinical condition of the patient was usually found: improvement of the clinical state was followed by decreased numbers of acantholytic cells.

Acantholytic cells were rounded or ovoid, having an enlarged, hyperchromatic, centrally or eccentrally situated nucleus. The cytoplasm was usually eosinophilic.

Neutrophilic polymorphonuclear leucocyte/acantholytic cell rosettes were sometimes seen in the smears, as well as eosinophil/acantholytic cell rosettes. Variable numbers of leucocytes (from 3 to 11) were seen surrounding the acantholytic cell.

Both binucleated epithelial and acantholytic cells and multinucleated giant cells were found in the smears. Multinucleated giant cells were sometimes seen engulfing cells resembling acantholytic cells.     

Clinical and cytological features of oral pemphigus

Nine patients with pemphigus vulgaris and one with pemphigus vegetans were studied. All ten patients had oral lesions, five of them skin involvement as well. The patients were followed up for variable periods at different intervals. At each visit, their clinical condition was assessed as mild, moderate, severe or remissive. At examinations, oral light microscopical smears and blood samples for indirect immunofluorescence were taken. From one patient with pemphigus vulgaris, smears from skin blister fluid were also studied. The highest intercellular antibody titers were found in severely affected patients, and those at the mild stage of the disease usually gave low or negative titers. A correlation between the number of acantholytic cells in smears and the clinical condition of the patient was usually found: improvement of the clinical state was followed by decreased numbers of acantholytic cells. Acantholytic cells were rounded or ovoid, having an enlarged, hyperchromatic, centrally or eccentrally situated nucleus. The cytoplasm was usually eosinophilic. Neutrophilic polymorphonuclear leucocyte/acantholytic cell rosettes were sometimes seen in the smears, as well as eosinophil/acantholytic cell rosettes. Variable numbers of leucocytes (from 3 to 11) were seen surrounding the acantholytic cell. Both binucleated epithelial and acantholytic cells and multinucleated giant cells were found in the smears. Multinucleated giant cells were sometimes seen engulfing cells resembling acantholytic cells.     

口腔天疱疮短期疗效影响因素分析

目的：对166例口腔天疱疮的基线资料和47例口腔寻常型天疱疮（PV）的治疗进行回顾性分析。方法：分析166例口腔天疱疮的性别比例,发病年龄,好发部位,临床类型;47例口腔PV按不同指标进行分组,评价激素短期治疗效果,并建立有关疗效的Logistic回归方程。结果：166例口腔天疱疮无性别差异,平均发病年龄49.8岁,口内最好发于颊部;口腔天疱疮的短期疗效与伴有皮肤损害,首次就诊,激素最大使用剂量等因素有关。结论：糖皮质激素联合免疫抑制剂治疗方案是目前治疗口腔天疱疮的主要方法,口腔伴皮肤损害者、非首次就诊者、激素最大剂量高者其短期疗效不佳。     

Immunodiagnosis of pemphigus and mucous membrane pemphigoid

Pemphigus and pemphigoid are two of a group of bullous diseases affecting oral mucosa and skin. Mucous membrane pemphigoid (MMP) comprises a heterogeneous group of disorders characterized by subepithelial separation and the deposition of immunoglobulins and complement along the basement membrane zone (BMZ). The target antigens in the epithelium and BMZ determine the nature of the condition, and recently there have been considerable improvements in our understanding of the BMZ antigenic composition. Pemphigus vulgaris (PV) is characterized by autoantibodies of the IgG isotype to the desmosomal glycoprotein desmoglein (Dsg) 3, whereas pemphigus foliaccus targets Dsg1, although at least 50% of PV patients have additional autoantibodies to Dsg1. The clinical phenotype appears to be determined by the relative amounts of Dsg1 and Dsg3. Patients with oral or mucosal PV have predominantly Dsg3 autoantibodies. The most frequently targeted antigen in MMP is bullous pemphigoid antigen 180 (BP180), although bullous pemphigoid antigen 230 (BP230), laminin 5, and beta 4 integrin are also involved. Circulating IgG and IgA antibodies may bind to different epitopes of BP180 namely the NC 16A domain or COOH -terminal domain. Pure ocular disease has been associated with IgA antibodies to a 45-kDa antigen and IgG antibodies to the 205-kDa antigen b4 integrin. The use of salt-split skin substrate enables differentiation between epidermal and dermal 'binders'. Since both the specificity and the antibody titer appear to have direct relationships with the disease severity, and a combination of clinical score and antibody titer provides valuable prognostic data, these investigations should be carried out on a more routine basis.     

Immunodiagnosis of pemphigus and mucous membrane pemphigoid

Pemphigus and pemphigoid are two of a group of bullous diseases affecting oral mucosa and skin. Mucous membrane pemphigoid (MMP) comprises a heterogeneous group of disorders characterized by subepithelial separation and the deposition of immunoglobulins and complement along the basement membrane zone (BMZ). The target antigens in the epithelium and BMZ determine the nature of the condition, and recently there have been considerable improvements in our understanding of the BMZ antigenic composition. Pemphigus vulgaris (PV) is characterized by autoantibodies of the IgG isotype to the desmosomal glycoprotein desmoglein (Dsg) 3, whereas pemphigus foliaceus targets Dsg1, although at least 50% of PV patients have additional autoantibodies to Dsg1. The clinical phenotype appears to be determined by the relative amounts of Dsg1 and Dsg3. Patients with oral or mucosal PV have predominantly Dsg3 autoantibodies. The most frequently targeted antigen in MMP is bullous pemphigoid antigen 180 (BP180), although bullous pemphigoid antigen 230 (BP230), laminin 5, and beta 4 integrin are also involved. Circulating IgG and IgA antibodies may bind to different epitopes of BP180 - namely the NC16A domain or COOH-terminal domain. Pure ocular disease has been associated with IgA antibodies to a 45-kDa antigen and IgG antibodies to the 205-kDa antigen b4 integrin. The use of salt-split skin substrate enables differentiation between epidermal and dermal 'binders'. Since both the specificity and the antibody titer appear to have direct relationships with the disease severity, and a combination of clinical score and antibody titer provides valuable prognostic data, these investigations should be carried out on a more routine basis.     

A determination of the range of oral conditions submitted for microscopic and direct immunofluorescence analysis

BACKGROUND: Direct immunofluorescence (DIF) testing is a useful adjunct for the diagnosis of immune-mediated oral vesiculobullous diseases, helping to identify separate, histologically similar, but prognostically different, conditions. It is unknown how often biopsy of these lesions yields positive DIF results. METHODS: A total of 270 consecutive archival cases submitted to a reference laboratory in Buffalo, New York, over a 2-year span were examined. These specimens were submitted to establish or rule out a diagnosis of a DIF-positive oral vesiculobullous disease. Demographic, clinical, and diagnostic information, based on conventional microscopic and DIF analysis, was tabulated. To assess the contribution of DIF to successful diagnosis, three pathologists examined the hematoxylin and eosin-stained slides of the known DIF-positive specimens without knowledge of the DIF results. RESULTS: Approximately 48% of the specimens demonstrated positive DIF findings and consisted of pemphigus vulgaris, mucous membrane pemphigoid, lichen planus, linear immunoglobulin A disease, and chronic ulcerative stomatitis. The remaining specimens had negative DIF findings and consisted of numerous non-specific inflammatory conditions. Of particular interest were several cases of epithelial dysplasia and squamous cell carcinoma. Of the DIF-positive cases, only pemphigus vulgaris could be diagnosed reliably by conventional microscopy alone. CONCLUSIONS: Approximately half of biopsies of oral conditions that clinically resembled typically DIF-positive vesiculobullous diseases did not yield positive findings on DIF testing. Instead, a wide range of oral diseases can mimic these lesions clinically. With the exception of pemphigus vulgaris, DIF is essential for establishing a definitive diagnosis for known DIF-positive diseases.     

Levels of serum immunoglobulin G specific to bacterial surface protein A of Tannerella forsythia are related to periodontal status

Background: Tannerella forsythia (Tf) is a Gram‐negative anaerobe implicated in the development of periodontal disease. Bacterial surface protein A (BspA) is a surface‐expressed and ‐secreted protein that is recognized as an important virulence factor of Tf. This study was undertaken to determine whether Tf BspA induces an antibody response in periodontal disease. We hypothesized that serum immunoglobulin (Ig)G antibody levels against BspA correlate with the disease of patients. Methods: Sera were obtained from 100 patients with cardiac disorders and periodontal disease and 73 patients who experienced myocardial infarction but were periodontally healthy. Sera samples were assayed for anti‐BspA antibody (total IgG and IgG subtypes) by enzyme‐linked immunosorbent assay (ELISA). Antibody levels were measured in ELISA units by using an arbitrary patient as a standard. Results: A negative correlation was found with BspA‐specific total IgG antibody titers and the severity of disease measured as the clinical attachment level (CAL) when healthy and diseased groups were analyzed separately (healthy group: [?0.23, correlation value] Student's t value [73 degrees of freedom] = 1.99; P = 0.05; diseased group: [?0.21] t [100 degrees of freedom] = 2.12; P = 0.03]). However, there was a positive correlation ([0.18 correlation value] Student's t value [173 degrees of freedom] = 2.39; P = 0.017) when healthy and diseased groups were combined. A strong positive correlation ([0.338 correlation value] Student's t value [173 degrees of freedom] = 4.69; P <0.0001) between the BspA‐specific IgG titers and periodontal probing depth was observed when healthy and disease groups were combined. Conclusions: Data demonstrated that antibodies to Tf BspA were elicited in patients with periodontal disease, and antibody levels were associated with the disease severity. Furthermore, data suggested that anti‐BspA IgG might have a protective function in periodontal disease by minimizing the loss of tooth attachment tissue.     

The effects of periodontal therapy on serum antibody (IgG) levels to plaque microorganisms

The influence of periodontal therapy on serum antibody titers to selected periodontal disease-associated microorganisms was assessed in 23 patients having chronic inflammatory periodontal disease (CIPD). The immunoglobulin G (IgG) titers were determined by the microELISA technique in serum samples obtained prior to treatment; following a hygienic phase which included scaling, root planing, and oral hygiene instruction; following surgical treatment; and one year and two years following hygienic phase (maintenance phase). Considerable individual variability existed in the magnitude of immune response to specific bacterial preparations. Significant reductions in the mean antibody titers were seen to A. viscosus, S. sanguis, F. nucleatum, S. sputigena, B. gingivalis, B. intermedius, B. melaninogenicus, T. vincentii, and T. denticola by the end of the second year of maintenance. There was no consistent response to Capnocytophaga. When individual patient responses were examined, 6 of the 23 were found to have elevated titers to at least one of the microorganisms in the interval between pretreatment and the end of the hygienic phase; however, in all but one case, the titers at the end of the second year of maintenance were below pretreatment levels. Antibody levels to bacteria such as S. sanguis were modified during therapy. This would indicate that immune responses to microbes not generally considered to be "periodontal pathogens" may be modified by adjuvant activity associated with subgingival plaque or changes in the environment of the sulcus and that subsequent changes in titer do not necessarily reflect a role of that microorganism in the disease process.