泪腺良性淋巴上皮病变免疫组织化学研究

目的 通过研究9种单克隆抗体在泪腺良性淋巴上皮病变中的表达,探讨其免疫病理本质及发病机制,为临床治疗及预后提供理论依据.方法 使用CD4、CD8、CD21、CD34、CD45、CD68、IgG、IgG4和增殖细胞核抗原这9种分子的单克隆抗体,应用免疫组织化学染色SP法对25例泪腺良性淋巴上皮病变的石蜡标本进行免疫组织化学染色.结果 B淋巴细胞表面抗原(CD21)、T淋巴细胞表面抗原(CD4、CD8和CD45)、黏附分子CD34及IgG在25例样本中均呈阳性表达,巨噬细胞表面抗原CD68、增殖细胞核抗原和IgG4这3种分子在25例样本中均有一定比例的阳性表达,表达率分别为12％、8％和80％.结论 泪腺良性淋巴上皮病变是一种主要以B淋巴细胞和T淋巴细胞激活为主要病理免疫基础的疾病,绝大多数属于IgG4相关性疾病的范畴,同时有发生恶变的潜能,临床治疗后的密切随访很有必要.     

Association analysis of BclI with benign lymphoepithelial lesions of the lacrimal gland and glucocorticoids resistance

AIM: To evaluate the relationship between gene polymorphism (BclI, ER22/23EK, N363S) and the occurrence, progression and sensitivity to glucocorticoid of lacrimal gland benign lymphoepithelial lesion (LGBLEL).METHODS: Clinical peripheral blood samples of 52 LGBLEL patients and 10 normal volunteers were collected for DNA extraction and polymerase chain reaction sequencing to analyze single nucleotide polymorphism (SNP) genotypes. The lacrimal tissues of LGBLEL were surgically removed and made into paraffin sections for subsequent hematoxylin-eosin (HE) and Masson staining analysis. The duration of disease and hormone use of LGBLEL patients from diagnosis to surgery were also analyzed. The Meta-analysis follows PRISMA guidelines to conducted a systematic review of human studies investigating the relationship between the NR3C1 BclI polymorphism and glucocorticoids (GCs) sensitivity.RESULTS: There was no association between ER22/23EK or N363S and the occurrence of LGBLEL or GCs sensitivity (P>0.05); BclI GC genotype was closely related to GCs resistance (P=0.03) as is the minor allele C (P=0.0017). The HE staining and Masson staining showed that the GC genotype of BclI remarkably slowed down the disease progression and reduced fibrosis (P<0.05), especially for GCs-dependent patients (P<0.0001). Meta-analysis showed that BclI was not significantly associated with GCs responsiveness.CONCLUSION: The LGBLEL patients who carry the NR3C1 BclI allele C may be more sensitive to GCs and associated with lower fibrosis and slower disease progression. The results may guide the clinical treatment strategy for the LGBLEL patients.     

B cell receptor signaling pathway involved in benign lymphoepithelial lesions of the lacrimal gland

AIM: To detect the expression of B cell receptor signaling pathway (BCRSP) in lacrimal gland benign lymphoepithelial lesions (LGBLEL). METHODS: Gene microarray was used to compare whole-genome expression in lacrimal gland tissues from LGBLEL patients to tissues from orbital cavernous hemangioma (control tissues). Expression of BCRSP was confirmed by polymerase chain reaction (PCR) and immunohistochemistry. RESULTS: The expression of 22 genes of the BCRSP increased significantly in LGBLEL patients. PCR analysis showed that CD22, CR2, and BTK were all highly expressed in LGBLEL tissues. Immunohistochemical analysis showed that CR2 protein was present in LGBLEL, but CD22 and BTK proteins were negative. CR2, CD22, and BTK were not observed in the orbital cavernous hemangiomas with either PCR or immunohistochemistry. CONCLUSION: BCRSP might be involved in the pathogenesis of LGBLEL.     

Different serum levels of IgG and complements and recurrence rates in IgG4-positive and negative lacrimal gland benign lymphoepithelial lesion

AIM: To analyze the differences in immune indicators and prognosis between IgG4-positive and negative lacrimal gland benign lymphoepithelial lesion (LGBLEL).METHODS: This was a single-center retrospective clinical study including 105 cases of IgG4-positive LGBLEL and 41 cases of IgG4-negative LGBLEL. Basic information, related indicators of peripheral venous blood samples using immunoscattering turbidimetry, treatment (partial surgical excision and glucocorticoid therapy) and prognosis (recurrence and death) were collected. Survival curves for recurrence were created using the Kaplan-Meier analysis. Univariate analysis and multivariate regression analysis were used to analyze prognostic factors.RESULTS: The mean age was 50.10±14.23y and 44.76±11.43y (P=0.033) in IgG4-positive and negative group respectively. The serum C3 and C4 was lower in IgG4-positive group (P=0.005, P=0.002), while the serum IgG and IgG2 was higher in IgG4-positive group (P=0.000 and P=0.008). Twenty-one cases had recurrence in IgG4-positive group and 3 cases recurrence in IgG4-negative group. The 5-year recurrence-free cumulative percentages of IgG4-positive group was 81.85%, and 83.46% in the IgG-negative group (P=0.216). The history of preoperative glucocorticoid therapy, serum C4, IgG1 and IgG2 were the factors affecting recurrence in IgG4-positive group, while serum C4, and IgG1 were the factors affecting recurrence of LGBLEL.CONCLUSION: Serum C4 and IgG1 are the factors affecting recurrence of LGBLEL, while the IgG4 does not affect recurrence of LGBLEL.     

IgG 亚型与泪腺良性淋巴上皮病变发病关系的研究

目的：检测泪腺良性淋巴上皮病变患者外周血IgG亚型的含量,探讨其与该病发病的关系。方法将2010年8月至2015年12月间来我院眼科就治,手术后病理组织学确诊的58例泪腺良性淋巴上皮病变患者为实验组,26例眼眶海绵状血管瘤患者为对照组,对临床资料及外周静脉血标本进行收集,采用酶联免疫吸附测定方法,分析血液中IgG亚型与泪腺良性淋巴上皮病变之间的关系。结果实验组与对照组外周血中IgG亚型含量的差异主要集中在IgG1、IgG2、IgG4和IgG（ P ＜0．05）,尤以IgG4和IgG差异最为显著。结论在诊断泪腺良性淋巴上皮病变中IgG4和IgG具有一定的参考价值。     

泪腺良性淋巴上皮病变的病理特点及IgG4和C3的表达

目的　探讨IgG4和C3在泪腺良性淋巴上皮病变发生发展中的作用。方法　收集2010年7月至2018年5月就诊于承德医学院附属医院眼科、解放军总医院眼科15例（15眼）泪腺良性淋巴上皮病变患者的泪腺肿物为试验组，10例（10眼）因其他疾病而行眶内容物摘除术的正常泪腺组织标本为对照组。采用HE染色法观察2组泪腺病理形态学变化及其组织病理学特点，采用免疫组织化学染色法检测2组泪腺组织中IgG4、C3的表达，并对二者表达相关性进行分析。结果　HE染色结果显示，对照组泪腺由正常腺泡及导管组成，导管上皮细胞排列整齐，细胞结构清晰，其间有少量的淋巴细胞；试验组泪腺组织中可见大量淋巴细胞、浆细胞浸润，淋巴滤泡形成，其间可见上皮-肌上皮岛结构改变，同时伴有不同程度纤维化。IgG4在试验组阳性表达面积为（30 934.80±16 057.17）像素，对照组阳性表达面积为（325.42±204.43）像素，试验组中明显高于对照组（t=-7.38，P=0.000）；C3在试验组阳性表达面积为（43 169.49±33 206.60）像素，对照组阳性表达面积为（323.24±271.29）像素，试验组中明显高于对照组，差异有统计学意义（t=-5.00，P=0.000）。试验组中IgG4与C3表达无相关性（r=-0.137，P=0.671）。结论　泪腺良性淋巴上皮病变患者的泪腺发生了明确的病理改变，这可能与IgG4和C3在泪腺中的高表达有关。     

泪腺良性淋巴上皮病变治疗方式的探讨

目的 探讨手术辅以激素治疗泪腺良性淋巴上皮病变（BLEL）的疗效.方法 回顾性系列病例研究.收集2010年8月至2013年2月间经病理组织学证实为泪腺BLEL患者的手术前后资料16例（16只眼）,分析和总结手术前后患者视力、眼压、眼睑肿胀、眼干及并发症.结果 16例患者术后眼睑肿胀明显减轻,眼睑位置及抬举功能正常;16例患者手术前后比较,视力升高1例,无变化15例;16例患者中仅有1例出现眼干症状,与术前比较术后眼干症状未见加重;16例患者仅1例发生手术后短暂的复视,1周后自行恢复.结论 以手术切除辅助激素治疗泪腺BLEL的方式是安全可行的.     

泪腺良性淋巴上皮病变的病因及发病机制

良性淋巴上皮病变以双侧或单侧泪腺、眼睑肿胀和（或）涎腺弥漫性无痛性肿大为主要临床表现.目前该病具体病因及发病机制尚未阐明.从病理学、感染学、免疫学、流行病学、分子生物学等多方面对该病的病因及发病机制进行总结,良性淋巴上皮病变的病因主要包括基底细胞增生浸润、病毒诱导、自身免疫功能障碍、性激素紊乱、蛋白紊乱、IgG4浸润等假说.这些假说都可以从不同侧面对良性淋巴上皮病变的病因及发病机制进行解释,但仍存一定争议,有待更深入研究,以便指导临床诊断及治疗.     

补体经典途径在泪腺良性淋巴上皮病变发生中的作用

目的：分析补体系统(CS)及其经典途径在泪腺良性淋巴上皮病变(LGBLEL)发病机制中的作用。

方法：采集LGBLEL患者和眼眶海绵状血管瘤(CH)患者的病变组织标本,使用蛋白质组学方法分析差异蛋白,而后采用逆转录-聚合酶链反应(RT-PCR)、免疫组织化学染色(IHC)和蛋白质印迹法(Western Blotting)验证CS信号通路中差异蛋白表达的变化,明确其在LGBLEL发病机制中的作用。

结果：蛋白质组学分析结果表明,相对于眼眶CH患者,LGBLEL患者病变泪腺组织中CS信号通路重要蛋白C3、C5、C9、C1q等表达均发生改变; RT-PCR检测结果显示,与眼眶CH患者相比,LGBLEL患者病变泪腺组织中C1qA、C5、C9 mRNA表达升高; 免疫组织化学染色结果显示,与眼眶CH患者相比,LGBLEL患者病变泪腺组织中C1qA、C3、C5、C9表达明显增多; Western Blotting检测结果显示,与眼眶CH患者相比,LGBLEL患者病变泪腺组织中C1qA、C3、C9蛋白表达水平明显升高。

结论：CS参与LGBLEL的发病机制,其经典途径可能是其发挥作用的途径之一。     

泪腺良性淋巴上皮病变临床表现及诊断思路的研究

目的探讨泪腺良性淋巴上皮病变（BLEL）的临床特点和诊断方法。方法回顾性系列病例研究。收集2010年8月至2011年10月间经病理证实的泪腺BLEL患者资料16例,分析和总结其临床特点、影像学表现及实验室检查结果。结果 16例患者中男：女=1：7;年龄为36～63岁,其中以41～50岁为多;单眼发病：双眼发病=1：7;上眼睑无痛性非充血性持续肿胀为其主要临床表现。眼眶MRI成像扫描显示,16例患者病变均累及眼睑和泪腺组织,受累泪腺明显肿大,T1WI和T2WI均呈等信号,增强扫描可见明显均匀强化,另外,3例患者眼外肌受累,1例颞肌受累,1例额神经受累;实验室检查仅1例患者丙肝抗体阳性,余未见异常。结论眼睑肿胀是BLEL的典型症状,受累泪腺以双侧居多;MRI成像检查在诊断BLEL中具有重要价值,但确诊该病需要有病理组织学检查结果的支持。     

泪腺良性淋巴上皮病变与眼眶海绵状血管瘤患者病变标本中差异表达基因的检测

背景 泪腺良性淋巴上皮病变是临床较少见的眼眶病,主要表现为双侧泪腺的对称性、无痛性肿大,其病因和发病机制至今尚未阐明. 目的 筛选泪腺良性淋巴上皮病变组织与眼眶海绵状血管瘤患者病变组织中的差异表达基因,从分子水平探讨泪腺良性淋巴上皮病变的发病机制.方法 收集2010年9月至2013年4月在首都医科大学附属北京同仁医院经病理学检查证实的泪腺良性淋巴上皮病变患者9例的病变标本,并收集同期眼眶海绵状血管瘤患者9例的组织标本作为对照.利用全基因组表达谱芯片技术和limma算法检测2个组患者组织标本中的差异表达基因,并采用实时荧光定量PCR法验证差异基因的表达,采用Fisher法和基因本体(GO)功能富集分析法对差异表达基因进行功能分析和信号通路分析,找到主要差异表达基因的功能群和信号通路. 结果 泪腺良性淋巴上皮病变患者与眼眶海绵状血管瘤患者中共筛选出5 260个差异基因,Fisher差异表达基因的功能显著性分析和信号通路分析显示,109个GO条目中的差异表达基因显著上调,101个GO条目中的差异表达基因显著下调,其中32个功能基因相关的信号通路显著上调,25个信号通路显著下调.GO分析显示,差异表达基因中补体受体介导的信号通路表达丰度最高,其次为T细胞信号通路和B细胞信号通路上调以及丝裂原激活蛋白激酶(MAPK)信号通路和转化生长因子-β(TGF-β)信号通路下调.实时荧光定量PCR结果显示,泪腺良性淋巴上皮病变组患者标本中TIPRL、TLR7和TLR10基因的相对表达量明显高于眼眶海绵状血管病组,差异均有统计学意义(Z=-2.03、-2.32、-2.32,均P＜0.05),与基因芯片检测结果一致.结论 人泪腺良性淋巴上皮病变组织与眼眶海绵状血管瘤病变组织中基因表达谱明显不同,这些差异表达基因除参与T细胞和B细胞信号通路的上调以及MAPK信号通路和TGF-β信号通路的下调外,还涉及补体系统的变化.泪腺良性淋巴上皮病变的发生和发展是多种基因和通路共同作用的结果.     

眶内泪腺良性肿瘤的CT诊断临床研究

目的:探讨CT对泪腺良性肿瘤的诊断价值。方法:回顾性分析30例经病理证实良性泪腺肿瘤临床资料和CT影像资料。结果:泪腺良性肿瘤患者30例中26例位于泪腺眶部,泪腺睑部4例。肿瘤椭圆形18例,不规则形4例,边缘不规则呈现分叶状或结节状8例。CT表现大多数为密度均匀,眼眶皮质受压,凹陷18例,骨皮质破坏伴硬化肥厚12例。结论:泪腺良性肿瘤CT有一定特征,诊断准确率高。     

姜黄素对糖尿病大鼠视网膜中巨噬细胞游走抑制因子表达的影响

目的检测巨噬细胞游走抑制因子（MIF）在糖尿病大鼠视网膜中的表达,探讨MIF在糖尿病视网膜病变（DR）中的作用及姜黄素早期干预对其表达的影响。方法对30只清洁级SD大鼠行腹腔注射质量分数1%链脲佐菌素（STZ）诱导建立糖尿病动物模型,选取其中的20只随机分为糖尿病组和姜黄素治疗组,各10只,另取10只正常匹配的大鼠作为正常对照组。姜黄素治疗组自STZ注射后3d起给予含羧甲基纤维素的姜黄素200mg/（kg.d）灌胃,共8周,正常对照组和糖尿病组给予等量的羧甲基纤维素灌胃。动物处死后收集各组大鼠眼球,苏木精-伊红染色观察大鼠视网膜组织的病理变化;采用ELISA法检测各组大鼠视网膜组织中MIF的表达;应用免疫组织化学法观察各实验组大鼠视网膜中MIF的表达及定位;透射电镜下观察大鼠视网膜超微结构的改变。结果与正常对照组比较,糖尿病组大鼠和姜黄素治疗组大鼠血糖明显升高,差异均有统计学意义（P〈0.01）;姜黄素治疗组大鼠血糖水平明显低于糖尿病组,差异有统计学意义（P〈0.01）。与正常对照组比较,糖尿病组和姜黄素治疗组大鼠体重明显下降,差异均有统计学意义（P〈0.01）;姜黄素治疗组大鼠体重明显大于糖尿病组,差异有统计学意义（P〈0.01）。糖尿病组大鼠视网膜变薄,但姜黄素治疗组大鼠视网膜接近正常。免疫组织化学检测表明,糖尿病组视网膜组织中含有大量MIF阳性细胞,姜黄素治疗组视网膜中MIF表达明显少于糖尿病组（P〈0.01）。ELISA法检测结果显示,糖尿病组MIF的表达较正常对照组显著升高（P〈0.01）;姜黄素治疗组MIF的表达较糖尿病组明显下降（P〈0.01）。透射电镜下糖尿病组大鼠视网膜光感受器内外节水肿,毛细血管内皮细胞和周细胞染色体边集;姜黄素治疗组大鼠视网膜超微结构的改变轻微。结论 DR与视网膜的炎性改变有关,视网膜组织中MIF的高表达参与早期DR的发生发展。姜黄素可下调MIF的表达,其早期干预可延缓DR的发病进程。     

Expression of macrophage migration inhibitory factor in Aspergillus fumigatus keratitis

AIM: To investigate the expression of macrophage migration inhibitory factor (MIF) and detect its role in the innate immune response of fungal keratitis (FK). METHODS: We collected the paraffin-embedded cornea tissues from 10 FK and 6 ocular trauma patients to explore the MIF expression by immunohistochemistry. Then we cultured telomease-immortalized human corneal epithelial cells (THCEs), stimulated by the hyphae suspension of Aspergillus fumigatus (A. fumigatus) to detect the change of MIF with or without the pretreatment of MIF inhibitor [4-Iodo-6-phenylpyrimidine (4-IPP)] by real-time polymerase chain reaction (PCR). The protein level of MIF was also tested by immunohistochemistry, and the level of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) mRNA were compared between normal, hyphae stimulated and 4-IPP pretreated groups by real-time PCR to study the influence of MIF on the expression of TNF-α and IL-6. Corneal severity of rats’ FK models was documented by clinical scores, and real-time PCR. Western blot and immunohistochemistry were used to test the expression of MIF, TNF-α and IL-6 in rats’ corneas. RESULTS: In the corneas of FK patients, there was much stronger expression of MIF than that in the normal group showed by immunohistochemistry. In cultured THCEs stimulated by A. fumigatus, the expression of MIF became stronger in both immunohistochemistry and PCR at 16, 24, 32 and 48h post infection (p.i.; P<0.01, P<0.01, P<0.01, P<0.05). After pretreated with 4-IPP, the expression of MIF reduced at 4, 8, 16h p.i. (P<0.05, P<0.05, P<0.05) and the downstream TNF-α and IL-6 decreased obviously (P<0.05, P<0.01). In rats with A. fumigatus keratitis, the relative mRNA and protein level of MIF increased than those in the normal group by PCR (at 1d: P<0.01, 3d: P<0.01, 5d: P<0.01), Western blot and immunohistochemistry. After blocked MIF with 4-IPP, the clinical outcomes of rat keratitis showed markedly reduced inflammatory response (P<0.01), with TNF-α and IL-6 decreased in accordance with those in THCEs by PCR (P<0.05, P<0.01). CONCLUSION: The expression of MIF increased significantly in FK patients, THCEs and rats stimulated by A. fumigatus. After blocked with 4-IPP, the expression of MIF reduced, and so did its downstream cytokines: TNF-α and IL-6. The inflammation reaction of the rats’ corneas lightened after pretreated with 4-IPP. MIF may play a role in the innate immune response of the corneal resistance against A. fumigatus.     

Unusual presentations of pleomorphic adenoma and adenoid cystic carcinoma of the lacrimal gland

Purpose: To report two atypical cases of pleomorphic adenoma and adenoid cystic tumours of the lacrimal gland. Methods: Two case reports are presented. The first is of a 65-year-old female with a long history of right hypoglobus with sudden recent worsening. Computed tomography (CT) showed a round, well-defined lesion in the fossa for the lacrimal gland with an anterior hypodense exten- sion suggestive of possible malignancy in a pleomorphic adenoma. The tumour in the second case, a 35-year-old male, was diagnosed after presentation following a relatively minor periorbital injury. The smooth rounded mass on CT scan was suggestive of a benign lacrimal gland tumour. Results: The lesion in case 1 was excised with a diagnosis of haemorrhage within a pleomorphic adenoma. The lesion in case 2 was excised with a diagnosis of adenoid cystic carcinoma of the lacrimal gland with pseudocapsule. Conclusions: Haemorrhagic cyst developing beneath the pseudocapsule of a pleomorphic adenoma should be considered in the differential diagnosis of secondary development of malignancy in a pleomorphic adenoma. Adenoid cystic tumours of the lacrimal gland can present with a pseudocapsule.     

眼眶良性淋巴上皮病变泪液分泌功能研究

目的 探讨眼眶良性淋巴上皮病变的泪液分泌功能变化情况.方法 对10例眼眶良性淋巴上皮病变患者的两眼术前术后泪液分泌功能变化进行1:1配对比较分析.结果 治疗1个月后,非手术眼反射性泪液分泌较手术眼及治疗前均有显著性增多差异有统计学意义(P＜0.05),手术眼反射性泪液分泌同治疗前相比差异无统计学意义(P＞0.05).结论 眼眶良性淋巴上皮病变对糖皮质激素治疗敏感,治疗后泪腺仍然可以恢复部分泪液分泌功能,手术时不宜切除过多的肿大泪腺.     

Correlation of computed tomographic and histopathologic features in malignant transformation of benign mixed tumor of lacrimal gland

A 65-year-old man had painless progressive proptosis of five years' duration in his left eye. Sudden increase of the proptosis required a lateral canthotomy. A lateral orbitotomy was performed and a large, encapsulated, globoid mass was totally excised. Histopathologic diagnosis was adenoid cystic carcinoma arising in and surrounding a benign mixed tumor of the lacrimal gland. The computerized tomographic features correlated quite accurately with the gross appearance and histopathologic findings observed in the lacrimal gland mass. The authors discuss the clinical and radiographic features that should alert the clinician to the possibility of malignant transformation in a benign mixed tumor of the lacrimal gland.