IFN-α在原发性舍格伦综合征患者唇腺组织中的表达

目的:检测原发性舍格伦综合征患者唇腺组织中IFN-α的表达水平,探讨IFN-α在原发性SS发病中的作用。方法:对37例原发性SS患者和24例非SS患者的唇腺组织进行IFN-α免疫组化染色,比较分析其在两组之间的差异。使用SAS6.12统计软件包对数据进行t检验。结果:37例原发性SS患者中,22例(59.46%)表达阳性,15例(40.54%)表达阴性;24例非SS组患者中3例(12.5%)表达阳性,其余21例(87.5%)均表达阴性。2组之间有显著差异。结论:IFN-α在原发性舍格伦综合征患者唇腺组织中存在表达异常。     

舍格伦综合征患者血清中骨桥蛋白等细胞因子的表达

目的:研究舍格伦综合征(SS)患者血清中骨桥蛋白(OPN)等细胞因子的含量及相互关系。方法:经病理学检查确诊的SS患者40例,其中原发性15型,继发性25例。酶联免疫吸附试验(ELISA)检测患者血清中骨桥蛋白,肿瘤坏死因子-α(TNF-α),转化生长因子-β1(TGF-β1)的含量。以20名健康志愿者作为正常对照。结果:与正常对照组相比,OPN和TNF-α水平在SS患者显著增加(P<0.05);继发性SS的TGF-β1血清水平显著高于原发性SS,但OPN和TNF-α的血清水平在原发性SS与继发性SS中没有显著差异(P>0.05)。结论:骨桥蛋白在原发性SS和继发性SS患者的血清中表达均高于正常对照组,但继发性SS较原发性SS表达更为显著。说明其表达与疾病的严重程度有关。继发性SS的TGF-β1血清水平显著高于原发性SS,说明继发性SS疾病的病理发生发展更为复杂。     

基质金属蛋白酶-3在舍格伦综合征患者唾液和唇腺中的表达

［摘要］ 目的 研究基质金属蛋白酶-3（matrix metalloproteinase-3,MMP 3）在舍格伦综合征患者唇腺和唾液中的表达水平,以及明确其表达水平与病理分级的关系。方法 采集35例舍格伦综合征患者（实验组）和20例黏液腺囊肿患者（对照组）的唇腺和唾液,应用酶联免疫吸附试验法（ELISA）检测唾液中MMP-3的表达水平,应用免疫组化方法观察唇腺中MMP-3的表达,使用图像分析软件（IPP 6.0）定量分析两组唇腺中MMP 3的表达强度,并将舍格伦综合征患者按照其病理分级分为低的1、2级和高的3、4级两组。结果 MMP-3在实验组唾液和唇腺中的表达高于对照组,差异有统计学意义（P<0.05）;MMP-3在病理分级高的患者唾液和唇腺中表达水平高于病理分级低的患者,差异有统计学意义（P<0.05）。结论 舍格伦综合征患者唇腺中的MMP-3可能参与了腺泡以及腺体基质的破坏,导致疾病的发生;唾液以及唇腺中MMP-3的表达水平可能成为舍格伦综合征的一项检测指标, 并为临床诊治提供参考依据。     

骨桥蛋白、TGF-β1及TNF-α在合格伦综合征中的表达及相关分析

目的:研究舍格伦综合征(Sj(o)gren's syndrome,SS)患者血清中骨桥蛋白(osteopontin,OPN)、TNF-α、TGF-β1三者间的相关性,为临床诊断提供依据.方法:选择经病理确诊的SS患者40例,其中原发性SS 15例,继发性SS 25例.ELISA检测患者血清中骨桥蛋白(OPN)、TNF-α、TGF-β1的含量,并利用SPSS 13.0软件包对数据进行统计学分析.结果:OPN和TNF-α在患者血清中的含量较正常人显著增高(P＜0.05),继发性SS的TGF-β1血清含量显著高于原发性SS(P＜0.05),TGF-p1的血清含量与正常对照组相比无显著差异.OPN与TGF-β1具有相关性.结论:OPN和TNF-d可作为SS检测的辅助手段,而TGF-β1可作为鉴别继发性SS和原发性SS的一种检测指标.OPN、TGF-β1在SS的发病过程中各自发挥作用的同时,还相互协调,相互牵制,充分体现了细胞因子网络的双向性、多样性和整体性.     

IFN-α在原发性合格伦综合征患者唇腺组织中的表达

目的：检测原发性合格伦综合征患者唇腺组织中IFN-α的表达水平，探讨IFN-α在原发性SS发病中的作用。方法：对37例原发性SS患者和24例非SS患者的唇腺组织进行IFN-α免疫组化染色，比较分析其在两组之间的差异。使用SAS 6．12统计软件包对数据进行t检验。结果：37例原发性SS患者中，22例（59．46％）表达阳性，15例（40．54％）表达阴性；24例非SS组患者中3例（12．5％）表达阳性，其余21例（87．5％）均表达阴性。2组之间有显著差异。结论：IFN-α在原发性合格伦综合征患者唇腺组织中存在表达异常。     

基质金属蛋白酶在舍格伦综合征患者唇腺组织中的表达及意义

目的:观察基质金属蛋白酶3、9(MMP-3、MMP-9)在舍格伦综合征患者唇腺中的定位及表达强度,探讨其在舍格伦综合征发病机制中的作用.方法:选取舍格伦综合征患者(病变组)和下唇黏液囊肿患者(对照组)唇腺组织标本各20例,通过免疫组织化学染色方法,观察MMp-3、MMP-9在2组唇腺组织中的表达;采用图像分析软件,定量分析MMP-3、MMP-9在2组唇腺组织中的表达强度,采用Mann-Whitney U检验进行统计学分析.结果:MMP-3、MMP-9表达于腺泡细胞及导管上皮细胞.MMP-9在病变组表达强度值为0.48±0.25,对照组为0.30±0.10,病变组表达显著高于对照组(P＜0.05);MMP-3在对照组无表达,在病变组强表达.结论:MMP-3、MMP-9在舍格伦综合征病变唇腺组织中的表达强度明显增强,特别是MMP-3;它们可能对淋巴细胞在腺实质细胞间的分布以及对腺体破坏发挥作用,参与腺泡及腺体基质的破坏过程,从而导致疾病的发生.     

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目的检测原发性干燥综合征（primary Sj？gren＇s Syndrome,pSS）患者唇腺组织中Caspase-3的表达水平,探讨其在pSS发病中的作用。方法分别采用免疫组化法和Western blot法对60例pSS患者和25名正常人唇腺组织中Caspase-3蛋白进行检测,分析并比较两组间的差异。使用SPSS17.0软件对数据进行分析。结果免疫组化结果显示：60例pSS患者唇腺组织中,8例Caspase-3表达阴性,52例表达阳性,阳性表达率86.7%;25名正常人唇腺组织中,Caspase-3阳性表达仅2名,阳性表达率为8.0%。两组间Caspase-3阳性表达率比较,差异有统计学意义（P〈0.05）。Western blot结果显示：Caspase-3蛋白在pSS患者唇腺组织中的表达（33.55±3.12）明显高于正常人唇腺组织（71.62±5.31）,差异有统计学意义（P〈0.05）。结论 Caspase-3在pSS患者唇腺组织中存在高表达。     

骨髓基质细胞抗原2在原发性舍格伦综合征中的表达及意义

目的: 检测骨髓基质细胞抗原2(bone marrow stromal antigen-2,BST-2)在原发性舍格伦综合征（primary Sjögren's syndrome,pSS）患者唇腺及外周血淋巴细胞中的表达,探讨BST-2对淋巴细胞增殖的作用,以期为原发性舍格伦综合征的发病机制研究及药物治疗靶点的开发提供理论基础。方法: 应用实时定量PCR技术检测30例pSS患者及30例对照患者外周血单个核细胞（peripheral blood mononuclear cells,PBMCs）和唇腺组织中BST-2的表达水平,并与临床指标进行相关性分析。采用免疫组织化学对唇腺组织中的BST-2进行定位及半定量分析。通过分选外周血中的淋巴细胞,实时定量 PCR明确BST-2异常表达的淋巴细胞亚群,唇腺免疫组织化学连续切片染色验证。慢病毒转染人B淋巴细胞系以过表达BST-2,CCK8检测BST-2对细胞增殖活性的影响,流式细胞术分析其对细胞凋亡的影响。采用SPSS 16.0软件包对数据进行统计学分析。结果: 与对照患者相比,pSS患者唇腺组织中BST-2表达升高,阳性细胞主要是浸润腺体的B淋巴细胞。同时,pSS患者外周血CD19⁺ B细胞BST-2与对照组相比表达升高。过表达B细胞中的BST-2可以促进细胞增殖,抑制细胞凋亡。结论: BST-2在pSS患者外周血及唇腺组织中表达升高,BST-2主要定位于B淋巴细胞。推测BST-2通过促进B细胞在腺体中的增殖和浸润,从而参与pSS的发生与发展。     

原发性干燥综合征患者唇腺组织中白细胞介素7的表达及临床意义

目的探讨原发性干燥综合征（pSS）患者唇腺组织中IL-7的表达及其与临床血清学参数间的关联。 方法收集2015年1月至2017年12月汕头市中心医院pSS患者135例,以及同期唇外伤正常对照个体12例。免疫组织化学法检测两组个体唇腺组织中IL-7的表达,分别评估腺上皮细胞与浸润淋巴细胞IL-7的表达情况。Pearson相关分析唇腺组织IL-7的表达与抗核抗体（ANA）、C3、C4、C反应蛋白（CRP）、IgG等血清学指标间的关联性。 结果IL-7在唇腺组织内淋巴细胞、腺泡导管及上皮细胞均可表达。浸润的淋巴细胞IL-7表达阳性率在pSS患者（88.14%,119/135）显著高于正常对照个体（8.3%,1/12）,差异有统计学意义（χ² = 46.82,P<0.001）。pSS患者唇腺组织中腺泡与导管上皮细胞与浸润淋巴细胞IL-7的表达相关（r = 0.27,P = 0.0012）,未发现有浸润淋巴细胞IL-7表达与血清指标间的相关性。 结论pSS患者唇腺组织中腺体上皮细胞、淋巴细胞IL-7异常高表达,且两者表达情况紧密相关,提示IL-7的表达可能参与了pSS腺体与淋巴细胞的相互作用,可作为pSS唇腺干预一个新的治疗靶点。     

腮腺磁共振造影在舍格伦综合征诊断中的价值

目的：探讨腮腺磁共振造影在舍格伦综合征（SS）诊断中的价值。方法：应用磁共振涎腺成像及磁共振造影（MRS）诊断腮腺表现为主的舍格伦综合征，在选择合理诊断标准的基础上，结合免疫学指标、病理学检查进行诊断。并与腮腺造影结果进行比较。结果：32例最终诊断为SS的25例患者中，23例MRS诊断出现典型的表现，即导管和腺泡不规则的扩张，并能排除腮腺占位性病变。敏感度：92％：特异度：71．4％；准确度：87．5％。伴有自身抗体出现的占68．％：唇腺活检符合SS表现的占72％。18例SS患者MRS与腮腺造影均有阳性表现，15例结果基本相同，占83.3％。结论：MRS能客观地反映舍格伦综合征患者受累腺体的病变情况和程度，是SS诊断和与腮腺肿瘤性疾病鉴别诊断的重要方法之一。     

Epithelial HLA-DR expression in labial salivary glands in Sjögren''s syndrome and nonspecific sialadenitis

The expression of the Class II major histocompatibility antigen HLA-DR was quantified in the epithelial cells of labial salivary glands from patients with Sjögrens Syndrome (SS) and compared with similar expression in glands showing non-specific sialadenitis and normal controls. In all glands more duct cells were positive than acinar cells but only in sialadenitis and SS was strong epithelial staining seen. The proportions of duct and acinar cells expressing HLA-DR were increased between normals and sialadenitis (P < 0.01) and between sialadenitis and SS (P < 0.001). However, for all cases increased expression of HLA-DR correlated to the increased proportion of inflammatory cells in the gland (P < 0.01). The results indicate that although HLA-DR is expressed on the epithelial cells in the glandular lesions of SS, this is not specific as it is also seen in sialadenitis. This supports the view that such expression is secondary to an inflammatory infiltrate and may not be of importance in initiating autoimmune tissue damage.     

Clinical and laboratorial profile and histological features on minor salivary glands from patients under investigation for Sj?gren’s syndrome

Diagnosis of Sjögren’s syndrome (SS) is complex and the usefulness of labial minor salivary glands biopsy in this process remains controversial. Objectives: to evaluate the clinical and laboratorial profile and histological features on labial minor salivary glands from patients under investigation of SS. Study Design: clinical charts from 38 patients under suspicion of SS and submitted to labial minor salivary glands biopsies were reviewed. Clinical and laboratorial data were retrieved from the clinical files and the HE-stained histological slides were reviewed under light microscopy. Results: mean age of the patients was 56.5 years and 97% were females; histological analysis showed that 42% of the cases showed ductal dilatation, lymphocytic foci were found in 52.6% and, from this group, 80% of the cases presented a foci/lobules ratio above 0.8. Acinar/ductal ratio was considered diminished in 39.5% of the samples. Thirty six (95%) and 32 (84%) patients, respectively, complained about xerostomia and xerophthalmia. A study of the time interval of the symptoms that led to SS investigation showed a mean of 116 months. Moreover, sixty-six percent of the patients had already been submitted to immunosuppressive therapy prior to the labial minor salivary gland biopsy. Age of the patients, scintigraphic alterations on salivary function, antinuclear factor (ANF), anti-Ro and anti-La did not show statistical significant association with the histological features. Lobules/foci ratio above 0.8 was the only histological parameter statistically associated with Sjögren’s syndrome diagnosis (p<0.0001). Conclusions: in the studied sample, lymphocytic foci on salivary glands were the only histological parameter associated to the diagnosis of SS. Early indication of labial minor salivary gland biopsy to patients under investigation of SS could limit the effects of immunosuppressive therapy on the histological features associated with the evolution of salivary gland involvement in SS. Key words:Sjögren syndrome, minor salivary glands, biopsy, lymphocytic foci.     

The minor salivary gland proteome in Sjögren's syndrome

Objective:  To identify the global protein expression (the proteome) in the minor salivary glands from primary Sjögren's syndrome (pSS) patients and non-SS controls.
Materials and methods:  Minor labial salivary glands were obtained from six pSS patients and from six age-matched non-SS controls, lysed in SDS buffer and pooled into two groups, respectively. The lysates were analysed by liquid chromatography electrospray ionization combined with tandem mass spectrometry. Also, the proteins were separated by two-dimensional polyacrylamide gel electrophoresis and protein spots were subjected to mass spectrometry.
Results:  Heat shock proteins, mucins, carbonic anhydrases, enolase, vimentin and cyclophilin B were among the proteins identified. The differences in the proteomes of minor salivary glands from pSS patients and non-SS controls were mainly related to ribosomal proteins, immunity and stress. Alpha-defensin-1 and calmodulin were among six proteins exclusively identified in pSS patients.
Conclusion:  We have identified several minor salivary gland proteins that may have implications for clarifying the SS pathophysiology. This experiment adds to the knowledge of proteins produced in salivary glands in health and disease, and may form the basis of further studies on biomarkers of prognostic and diagnostic value.     

Quantification of plasma cells in labial salivary glands: increased expression of IgM in Sjögren''s syndrome

Plasma cells expressing IgG, IgA and IgM were quantified in labial salivary glands from patients with Sjögren's syndrome (SS) and compared with glands showing non-specific inflammatory changes and normal controls. In all glands the predominant isotype was IgA but in SS there was a significant increase in both the number and proportions of IgG and IgM positive cells (P> 0.002). In particular, all SS cases contained greater than 10% IgM positive cells (mean = 26.8± 15.5). The results suggest that accumulation of IgM positive plasma cells may be a specific finding in SS and support the concept that the glandular lesions may be a site of B-cell clonal expansion. Since most B-cell hyperproliferative states in SS, including lymphoma, are associated with synthesis of IgM simple quantification of plasma cells may have important diagnostic and prognostic significance.     

唾液析晶形态试验在诊断舍格伦综合征中的应用

目的探讨唾液析晶形态试验(salivaferningtest,SFT)在舍格伦综合征(Sjgren′ssyndrome,SS)诊断中的价值。方法共收集78例确诊为SS的患者(采用欧洲标准),在进行唇腺活检之前取非刺激性新鲜唾液,光镜下观察其结晶形态,根据Maragou标准记录分型,根据Tarpley标准记录唇腺活检结果，并比较两种方法的阳性检出结果。同期随机选择80名健康人设为对照组，观察唾液析晶形态，检测SFT的敏感性和特异性。结果①SFT具有较高的敏感性（89.74%）和特异性（83.75%）。②SFT阳性结果在SS病例组和对照组比较差异有显著性（P<0.01）。③SFT和唇腺活检在病例组的阳性检出结果差异无显著性(P>0.05)。结论SFT对SS患者的早期诊断及治疗具有重要的参考价值。     

The salivary gland component of Sj?gren's syndrome: an evaluation of diagnostic methods

The diagnostic value of sialography, scintigraphy, sialometry, and labial salivary gland biopsy--as indicators of salivary gland dysfunction in Sj?gren's syndrome (SS)--was evaluated in 41 patients suspected of having SS. In about 70% of the cases, each of the four examinations showed changes that could indicate a salivary gland component of SS. However, the disease specificity of sialographic and scintigraphic examinations was low. Although the specificity of the labial salivary gland biopsy examination is considered high, our study revealed some cases in which a focus score of 1 to 3 was not accompanied by abnormal changes in either sialometry, sialography, or scintigraphy. At a focus score of more than 3, a fair amount of agreement on the results of the various diagnostic procedures was found. Because labial salivary gland examination shows only minor salivary glands and sialography and scintigraphy include only major salivary glands, variance between different diagnostic procedures is expected. This indicates a potential diagnostic role for sialographic and scintigraphic examination when the labial salivary gland biopsy is insufficient.     

Follicular dendritic cells confirm lymphoid organization in the minor salivary glands of primary Sjögren’s syndrome

Background: Sjögren’s syndrome (SS) is an autoimmune chronic inflammatory disorder affecting the salivary and lacrimal glands. The aim of this study was to explore immunophenotypic features of chronic inflammatory reactions in the minor salivary glands in patients with primary SS (pSS). Methods: Formalin‐fixed, paraffin‐embedded labial minor salivary gland tissue sections from randomly selected patients with pSS (n = 60) were investigated for the expression of CD21, CD23, CD35 and IgD by immunohistochemistry. Results: Based on the distribution and staining pattern of CD21, CD23, CD35 and IgD in lymphoid aggregates, several stages of chronic inflammatory reactions were observed. In 12/60 (20%) patients, lymphoid infiltrates with germinal centre (GC)‐like features such as extensive networks of CD21‐, CD23‐ and CD35‐positive cells were observed in the minor salivary gland tissue. Smaller networks and /or focal infiltrates with scattered CD21⁺, CD23⁺ and CD35⁺ cells were observed in the remaining 48/60 (80 %) cases. When dividing patients according to the presence (GC+) or the absence (GC?) of GC in the minor salivary glands, the mean focus score was significantly higher in the GC+ patients (P < 0.05). Double staining of the minor salivary glands revealed focal infiltrates with follicular dentritic cell networks and B cells resembling normal GCs in tonsillar tissue. Conclusion: A particular cellular profile was demonstrated in a sub‐group of patients with pSS and could be linked to serological aberrations. These findings warrant further prospective studies.     

Vascular endothelial growth factor (VEGF) and chronic graft-versus-host disease (cGVHD) in salivary glands of bone marrow transplant (BMT) recipients

BACKGROUND: The purpose of this study was to compare the immunolocalization of vascular endothelial growth factor (VEGF) in salivary glands of bone marrow transplant (BMT) recipients with normal controls and between the different stages of chronic graft-versus-host disease (cGVHD). In addition, the impact of the immunolocalization of VEGF on the survival rate of BMT patients was investigated as well. METHODS: Labial salivary glands obtained at the day 100+ from 36 consecutive patients, who underwent BMT, were included in the study. The streptavidin-biotin-peroxidase complex stain was used to detect VEGF in the salivary glands. Time of death after BMT was displayed by means of the Kaplan-Meier method for the following parameters: age and gender of the patients, donor gender, acute GVHD, cGVHD staging at the labial salivary glands, primary disease, platelet and neutrophils counts on day of biopsy, stem cell, oral mucositis, parenteral nutrition, oral lichenoid lesions of GVHD, conditioning regimen and immunolocalization degree of VEGF in labial salivary glands. The data were initially analyzed by means of the log-rank test and then included in the Cox's proportional hazard model. RESULTS: No differences on the immunolocalization of VEGF in the labial salivary glands of BMT recipients and control group or between the different stages of glandular cGVHD were noted. Both univariate and multivariate analysis of the survival rate showed significance of 5% only for platelet count over 100 x 109/l on the day of biopsy and male donor gender. CONCLUSIONS: Platelet count over 100 x 109/l and male donor gender are positive predictive factors on the survival rate after BMT. In addition, the immunolocalization of VEGF in salivary glands is not altered in BMT recipients at day 100+ and is not influenced by the stage of cGVHD.     

An association between salivary gland disease and serological abnormalities in Sjögren's syndrome

In the labial salivary glands (LSGs) of 16 primary and 18 secondary Sjögren's syndrome (SS) patients, infiltrating lymphocytes were histologically and immunohistochemically examined: also, the serum levels of rheumatoid factor, antinuclear antibodies, anti-DNA antibodies, anti-SS-A and anti-SS-B antibodies, and immunoglobulins (including IgG, IgM and IgA) were all assayed. An immunohistochemical analysis of the lymphocyte subsets in LSGs revealed that severe lymphocytic infiltration was frequently accompanied by marked B cell accumulation both in primary and secondary SS patients. Furthermore, local B cell accumulation was also closely associated with elevated levels of anti-SS-A and anti-SS-B antibodies and IgG, and this association was statistically significant in the group with primary SS but not in the group with secondary SS. Thus, local lymphocytic infiltration, especially B cell accumulation, in the salivary glands is suggested to be involved in serological abnormalities in primary SS. while complicated autoimmune diseases other than SS may also be involved in serological abnormalities in secondary SS.