Immunocytochemical study of granular duct cells with a hormonally enhanced granular cell phenotype in the mouse parotid gland

In the parotid glands (PGs) of intact male mice (12 weeks of age, ICR strain), immunofluorescence labels for a true tissue kallikrein, mK1, and for nerve growth factor (NGF) were recognized through the subluminal edges of the striated duct (SD) segments and interlobular duct segments. Because of their small size, secretory granules were not detectable by light microscopy in any of the duct cells. Full-fledged granular cells, containing large secretory granules that were visible by light microscopy, were induced in the SD segments of male mice after the injection of 5α-dehydrotestosterone (DHT) and triiodothyronine (T₃), given either alone or in combination every other day for 2 weeks. A stronger effect was detected in the mice that were concomitantly injected with DHT and T₃, and more abundant, fully developed granular cells appeared in the SD segments of these mice. These full-fledged granular cells were immunoreactive for mK1, NGF, and epidermal growth factor, but not for renin. The present results indicate that some of the SD cells with small granules in the mouse PG can develop a granular cell phenotype, producing more kinds of growth factors, as a result of the actions of androgen and thyroid hormone.     

Morphologic changes in the granular convoluted tubule cells of the mouse submandibular gland following hypophysectomy and hormonal replacement

 Morphological changes in the granular convoluted tubule (GCT) cells of the male mouse submandibular gland (SMG) were examined following hypophysectomy and hormonal replacement. Semithin sections stained with Heidenhain's iron hematoxylin showed that hypophysectomy severely regressed the GCT phenotype. Although only a few dispersed cells containing secretory granules were observed in the GCT segments under a light microscope, electron microscopy revealed that many regressed cells continued to constitutively elaborate apical secretory granules (although they were very small) and contained a euchromatic nucleus at the center of the cell, poor rough endoplasmic reticulum (RER) and Golgi apparatus in the perinuclear region, and well-developed basal infoldings. These findings suggest that hypophysectomy resulted in atrophy of GCT cells, but that they retained evidence of being secretory cells. 5α-Dihydrotestosterone (DHT); 3,5,3′-triiodo-l-thyronine (T₃); and dexamethasone (Dex) each enhanced the GCT phenotype of hypophysectomized males to some degree. Combined hormonal replacement with DHT + T₃ in hypophysectomized males restored a nearly normal male GCT phenotype with a full complement of secretory granules and rare basal infoldings, whereas T₃ alone induced a normal female-like GCT phenotype, with considerably abundant secretory granules and the usual short basal infoldings, in hypophysectomized male glands. Furthermore, Dex was found to synergistically enlarge secretory granules when administered with T₃ and/or DHT, although it was only weakly effective in enhancing the GCT phenotype when used alone. Taken together, the above findings confirmed that the GCT phenotype of the mouse SMG is regulated by the synergistic action of pituitary-dependent hormones. Received: October 1, 2001 / Accepted: May 13, 2002     

A型肉毒素对腮腺分泌功能影响的研究

目的 探讨大鼠腮腺注射A型肉毒素(botulinum toxin type A,BTX-A)后组织学和免疫变化的机制.方法 18只雌性Wistar大鼠分为A、B组.A组动物(n=6)右侧腮腺内注射0.1 ml的生理盐水;B组动物(n=12)右侧腮腺注射BTX-A(2.5 U/0.1 ml).两组动物分别在注射后第7、12、35天处死并行组织学及免疫组化观察.结果 A组、B组注射后第7天部分腺泡细胞轻度萎缩,血管活性肠肽免疫反应(vasoactive intestinal polypeptide immunoreaction,VIP-IR)阳性纤维密度显著减低(n=4,P＜0.05);12 d萎缩的腺泡细胞和腺管明显增多,VIP-IR阳性纤维密度减低更明显(n=4,P＜0.001);35 d腺细胞和VIP-IR纤维密度与生理盐水组基本相同.结论 局部注射BTX-A可以导致大鼠腮腺腺泡细胞和腺管短时性萎缩,VIP-IR表达减少,提示BTX-A通过抑制调控腮腺分泌的血管活性肠肽(vasoactive intestinal polypeptide,VIP)释放导致腺组织非损伤性萎缩,而产生腺体分泌减少.     

腮腺脂肪瘤样多形性腺瘤的诊断与治疗分析

目的 探讨腮腺脂肪瘤样多形性腺瘤的病理特点、免疫组织化学特点、诊断标准和有效治疗手段.方法 腮腺脂肪瘤样多形性腺瘤患者2例,行瘤体加瘤周5 mm部分腺体摘除术,术后肿物标本行常规病理学和免疫组织化学检查;患者术后随诊26 ～ 36个月,观察预后情况.结果 常规病理镜下见肿瘤组织包膜完整,肿瘤实质由黏液软骨样基质、脂肪组织及肌上皮细胞构成;脂肪组织弥漫均匀分布于瘤组织内,呈蜂窝状结构.免疫组织化学检查发现肿瘤组织发现,胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)在肌上皮细胞胞浆阳性表达;S-100蛋白表达阳性,主要在脂肪细胞表达;上皮膜抗原(epithelial membrane antigen,EMA)阳性,胞膜表达,主要在内层导管细胞表达;细胞角蛋白AE1/AE3阳性;细胞核增殖抗原Ki-67在细胞核的阳性表达率1％;患者术后随诊期内未见肿物复发.结论 病理和免疫组织化学检查是确诊腮腺脂肪瘤样多形性腺瘤的依据,手术是治疗本病的有效手段.     

Bcl-2及Bax在大鼠腺体萎缩过程中的表达及临床意义

目的：：探讨大鼠腮腺主导管结扎后B cell lymphoma 2(Bcl-2)、Bcl-2-associated X protein(Bax)在腺体萎缩过程中的表达及临床意义。方法：对72只Wistar大鼠进行不同时间点腮腺主导管结扎建立腮腺萎缩模型;分别于术后1、3、5、7、14、21、30、60、90、150、180 d取材,采用免疫组织化学染色法检测Bcl-2、Bax在不同时间点的表达及分布情况。结果：主导管结扎后7 d多数腺泡细胞萎缩消失。Bcl-2和Bax在正常腺体组织中呈低水平表达。主导管结扎后不同时间点Bcl-2和bax在腺体中呈高表达,3 d时Bax表达达到峰值(1.99±0.10),21 d时Bcl-2的表达达到峰值(3.02±0.10),随后Bcl-2及Bax表达均降低。 Bcl-2/Bax比值在1~21 d升高,随后降低并趋于稳定。结论：腮腺主导管结扎后Bax、Bcl-2的表达与腮腺萎缩过程相关。     

腮腺细胞程序性死亡分子5的表达及其与细胞凋亡关系的动物实验

目的 探讨大鼠腮腺主导管结扎后细胞程序性死亡分子5(programmed cell death5,PDCD5)的表达及其与腮腺细胞凋亡的关系,以期了解PDCD5在腮腺萎缩中的作用.方法 对66只Wistar大鼠建立腮腺主导管结扎动物模型,分别在结扎0、1、3、5、7、14、21、30、60、90和120 d(每个时间点6只大鼠)获取腮腺标本,免疫组化法检测腮腺细胞中PDCD5的表达,脱氧核糖核酸末端转移酶介导的原位缺口末端标记法检测腮腺细胞的凋亡.结果 PDCD5蛋白在正常的腺泡细胞和导管细胞中呈低水平表达,均匀分布于胞质.腮腺主导管结扎后,PDCD蛋白的表达水平显著上调,在结扎后3d腺泡细胞中PDCD5的吸光度值达峰值(1.261±0.048),胞质和胞核中均有表达.在结扎后1、3d,腺泡细胞中PDCD5的吸光度值均显著高于导管细胞,二者差异有统计学意义(P＜0.01).细胞凋亡检测结果显示,结扎后3d腮腺细胞凋亡指数显著增高,腺泡细胞凋亡指数[(21.750±0.119)％]显著高于导管细胞[(5.720±0.205)％],二者差异有统计学意义(P＜0.01).PDCD5蛋白的表达水平与腮腺主导管结扎诱导的细胞凋亡呈显著正相关(r=1,P=0.003).结论 大鼠腮腺主导管结扎后PDCD5的表达与腮腺萎缩过程密切相关,PDCD5在腮腺结扎后的细胞凋亡途径中可能发挥重要作用.     

腮腺腺淋巴瘤中5种免疫组化标志物的定位研究

目的：检测腺淋巴瘤和腮腺正常组织中癌胚抗原(carcinoembryonic antigen,CEA)、抗糜蛋白酶(α_1- antichymotrypsin,α_1-ACT)、抗胰蛋白酶(α_1-antitrypsin,α_1-AT)、乳铁蛋白(lactoferrin,LF)、转铁蛋白(transferrin,TF)的表达及定位,对腺淋巴瘤上皮的组织发生学进行探讨。方法：应用免疫组织化学技术(SABC法)检测36例腺淋巴瘤和正常腮腺CEA、α_1-ACT、α_1-AT、LF和TF的表达,进行阳性着色评价和定位分析。统计分析采用SPSS10．0统计软件包进行数据处理。结果：正常腮腺中腺泡细胞和闰管细胞对CEA、α_1-ACT、LF和TF呈阳性反应；纹管细胞中CEA、LF和TF阳性表达,未见α_1-ACT和α_1-AT着色；排泄管腔细胞对LF及CEA着色,排泄管基底细胞仅对CEA阳性；肌上皮细胞仅对TF阳性。腺淋巴瘤上皮成分中,TF主要分布于基底细胞,CEA、α_1-ACT、LF和α_1-AT主要分布于腔细胞。结论：腺淋巴瘤上皮成分中2层细胞形态结构与免疫表达不同,功能分化有差异；腺淋巴瘤上皮成分和腮腺腺泡细胞、闰管细胞有相似的免疫特征,提示两者间可能存在组织发生学联系。     

Thyroid hormone regulation of granular intercalated duct cells in the submandibular glands of female mice

Intercalated ducts (IDs) in the submandibular glands (SMGs) of mice exhibit a sexual dimorphism, in which a few cells in the IDs of females, but not of males, possess secretory granules. The effects of a hypophysectomy (Hypox) followed by the administration of triiodo-l-thyronine (T₃) on such granular intercalated duct (GID) cells in the female gland were histologically examined. Semithin sections stained with Heidenhain's iron hematoxylin revealed that Hypox resulted in the complete disappearance of the GID cells. In Hypox females, electron-microscopy examination of the ID cells whose localization corresponded to that of the GID cells in normal females showed that these cells had a pale, centric nucleus, poorly developed rough-surfaced endoplasmic reticulum (RER) and Golgi apparatus, and no secretory granules. When T₃ (1mg/kg body weight) was given to Hypox female mice every other day for 2 weeks, the GID cells reappeared in most of the ID segments. Electron microscopy revealed that these cells had abundant secretory granules in their apical cytoplasm, a nucleus located near the base of the cell, and layered cisterna of RER and segments of Golgi apparatus in the perinuclear cytoplasm. The localization and distribution of the GID cells in the T₃-treated Hypox females were almost the same as those in normal females. Taken together, these results suggest that thyroid hormones upregulate the GID phenotype, and that thyroid hormones are essential for the exocrine activities of GID cells.     

儿童腮腺区Castleman病1例报告及文献复习

本文报告1例发生于儿童腮腺区的Castleman病病例。该患儿临床表现为右侧腮腺区无痛渐大性肿物,无明显其他伴随症状。采取手术切除,术后病理诊断为Castleman病(局灶性,透明血管型)。结合相关文献进行探讨,以进一步提高对发生于儿童腮腺区Castleman病的认识。     

The rare entity of cystadenocarcinoma (CAC) in parotid gland: A single-center experience

Purpose

Cystadenocarcinoma (CAC) is an extremely rare disease in parotid gland. This study aimed to identify the clinical characteristics of CAC, and the therapeutic options for its treatment. An attempt was also made to identify postoperative recurrence-related risk factors.

Immunocytochemical localization of mK1, a true tissue kallikrein,in the mouse parotid gland: sexual dimorphism and effects of castration and hypophysectomy

In the normal parotid glands of mice at 12 weeks of age, mK1, a true tissue kallikrein, was detected at the apical rim of the striated ducts (SDs). Sexual dimorphism in the immunostaining intensity in parotid glands was seen, i.e., immunostaining was more intense in males than in females. Under electron microscopy, secretory granules, being small in size, and condensed at the subluminal cytoplasm, were labeled with immunogold particles showing the presence of mK1. These secretory granules were rather abundant and large in males. Castration in males reduced the immunoreactivity of mK1 in the SD cells because of a decrease in the number and size of secretory granules as revealed by electron microscopy. Hypophysectomy in male mice resulted in considerable loss of immunoreactivity for mK1, which was characterized under electron microscopy by complete disappearance or significant reduction of secretory granules in many SD cells. These results suggest that mK1 expression in the SD cells of murine parotid glands is regulated by pituitary-dependent hormones, and sexual dimorphism of mK1 expression is regulated by androgens.     

Ultrastructural study of hormonally responsive striated duct cells in the mouse sublingual gland

In semithin sections stained with Heidenhain's iron hematoxylin, a few scattered granular cells were observed in the striated ducts (SDs) of sublingual glands (SLGs) of the mouse; they were seen normally only in the glands of adult males. However, it was shown by electron microscopy that many SD cells, other than these granular cells, had apical secretory granules, thus forming a granular striated tubule (named the GST in this study) in a portion of SD segments in both sexes. Sublingual GST cells had very small dense secretory granules near the apical surface, with the nucleus in the apical one-third to one-half of the cell; small Golgi apparatus; sparse rough endoplasmic reticulum (RER); and well-developed basal infoldings. However, some granular cells in male GSTs had abundant large dense secretory granules in the apical two-thirds of the cell, a basal nucleus, and modest basal infoldings. Such granular SD cells disappeared after castration in males. Granular SD cells could be induced in the GSTs of females by the injection of 5α-dihydrotestosterone (DHT), triiodothyronine (T₃), and/or dexamethasone (Dex); given simultaneously, these hormones acted synergistically in this induction. These results indicate a close similarity between the duct systems of the SLG and those of the submandibulan gland (SMG) of the mouse: granular SD cells of the GST in the SLG resemble GCT cells in the SMG in expressing some of the same biologically active polypeptides, in being sexually dimorphic, and in being under the same multihormonal regulation. Received: October 19, 2000 / Accepted: April 24, 2001     

Induction of Sca-1 in the duct cells of the mouse submandibular gland by obstruction of the main excretory duct

The effect of ligation of the main excretory duct (MED) of the mouse submandibular gland (SMG) on the expression of Sca-1, a stem cell antigen, was examined by Western blotting and immunohistochemistry. By Western blotting, the expression of Sca-1 with a molecular weight of 18 kDa was identified in the normal gland. At 1 day post-ligation, the expression level of Sca-1 was strongly increased in the experimental gland and weakly in the contralateral gland, and such expression in both glands decreased at 6 days. By immunohistochemistry, Sca-1 was detected weakly in the apical membrane of excretory duct (ED) cells of the SMG under the normal condition. By duct ligation, Sca-1 became expressed strongly in most cells of the two major duct systems, i.e., the striated duct (SD) and granular convoluted tubules (GCT), but was not detected in the acinar (Ac) cells. By fluorescence-activated cell sorter (FACS) analysis, the number of side population (SP) cells in this gland was found to be increased by ligation. These results imply that Sca-1-positive cells may have a role in the duct cell proliferation in the regeneration step elicited by MED ligation-induced injury.     

Alterations of antioxidant biomarkers and type I collagen deposition in the parotid gland of streptozotocin-induced diabetic rats

Background and objective

Acarbose is a competitive inhibitor of intestinal alpha-glycosidases that slows the breakdown of sucrose and starch, thereby reducing glucose and fructose absorption. The aim of this study was to evaluate the effect of acarbose treatment on antioxidant parameters and deposition of type I collagen in the parotid glands of diabetic rats.

Methods

Diabetes mellitus was induced by intravenous injection of streptozotocin, and rats were divided into four groups: non-diabetic (NDM), diabetic (DM), diabetic treated with 25 mg/kg acarbose (DMA) and non-diabetic treated with acarbose (NDMA). Changes in enzymatic antioxidant systems, such as the activity of SOD and GPx enzymes, were evaluated, and the specific staining pattern of the type I collagen fibres was investigated in the rat parotid glands.

Results

The DM group presented high levels of SOD and GPx enzymes, which were reduced by acarbose treatment. Tissue damage, which was indicated by an increased MDA concentration in the parotid glands of rats in the DM group, was also reversed in the DMA group. Moreover, type I collagen fibres from DM rats were more intensely stained than those of NDM rats. Acarbose treatment was effective in decreasing collagen deposition, which was shown by a decrease in staining intensity of approximately 25%.

Conclusions

These results suggest that the diabetic state influences the type I collagen concentration in the parotid glands of rats. In addition, acarbose treatment was helpful in preventing the deposition of such fibres, as well the increase in oxidative stress induced by hyperglycemia.     

腮腺黏膜相关淋巴瘤临床及病理研究

目的：研究腮腺黏膜相关淋巴瘤(Mucosa—Associated Lymphoid Tissue Lymphoma，MALToma)病理诊断特征及临床治疗。方法：收集并分析临床资料，利用HE染色，LCA、CD20、UCHL-l，κ链、λ链免疫组织化学标记观察10例腮腺MALToma，结果：腮腺MALToma组织学具有中心细胞样细胞弥漫浸润，浆细胞分化，淋巴滤泡破坏以及形成上皮-肌上皮岛淋巴上皮病灶的特点。免疫组化证实其来源为B细胞淋巴瘤。结论：①本组ll例原发腮腺恶性淋巴瘤均属于MALToma。②部分(36％)伴良性淋巴上皮病及舍格林综合症提示，本组MALToma可能与自身免疫性疾病有关。③手术切除辅以适当化疗可获较好疗效。     

Immunohistochemical analysis of the proliterative capacity of duct and acinar cells during ligation-induced atrophy and subsequent regeneration of rat parotid gland

To study the proliferative capacity of salivary gland, an animal model of regeneration was developed. A clamp, which induced atrophy in parotid gland by obstructing the main excretory duct but allowed restoration of duct patency following removal, was implanted in a series of rats. When it was removed (Day 7), the weight of the glands was reduced by 50% and acinar cells had decreased from 93.8% to 8.2% of total cell population. Regeneration occurred rapidly following removal of the clamp. The number and location of cycling intercalated, striated, and excretory duct cells and acinar cells were monitored using an antibody to proliferating cell nuclear antigen (PCNA). All cell types were induced to cycle but the predominant cell to cycle was the acinar cell. During regeneration the number of PCNA+ acinar cells increased 38.7–fold from steady-state values. Results demonstrate that acinar cells have a significant potential for cycling, contrary to current histogenetic theories of salivary gland tumourigenesis which exclude acinar cells as potential progenitor cells on the grounds of their putative limited cycling capacity.     

Mucoepidermoid carcinoma-associated expression of MUC5AC,MUC5B and mucin-type carbohydrate antigen sialyl-Tn in the parotid gland

ObjectivesThe aberrant expression of mucins and mucin-type carbohydrates has been described in many types of cancer, including mucoepidermoid carcinoma (MEC), a malignant salivary gland tumor. In this study, we examined the aberrant expression patterns of mucins (MUC1, MUC4, MUC5AC and MUC5B), simple mucin-type carbohydrate antigens (Tn, sialyl-Tn and T) and mature carbohydrate antigens (Lewis^a and sulfo-Lewis^a antigens) in MEC originating from the parotid gland, which normally does not secrete mucins.DesignWe conducted an immunohistochemical study to investigate the presence of mucins and carbohydrates in 24 MEC samples originating from the parotid gland and in surrounding normal tissue of the same gland in comparison 6 samples of normal salivary glands. The expression levels were compared with respect to the histological grading. Furthermore, 24 MEC samples from non-parotid salivary glands were included.ResultsWe observed loss of topology of membrane-bound MUC1 and MUC4, and de novo expression of MUC5AC, MUC5B and sialyl-Tn in MEC that originated in the parotid gland. Furthermore, mucins MUC1, MUC4 and carbohydrate antigens Tn, sialyl-Tn, T, Lewis^a and sulfo-Lewis^a were overexpressed in MEC samples compared to surrounding normal salivary gland tissues. MUC1 was expressed in both low- and high grade MECs, whereas MUC4 was not expressed in high grade MECs of the parotid gland.ConclusionDuring the development of MEC in the parotid gland, the genes for gel-forming secretory mucins are switched on. Besides these MEC tissues overexpress short oligosaccharides, suggesting that the glycosylation machinery is altered.     

肌上皮细胞在大鼠腮腺萎缩过程中的转归

目的 研究大鼠腮腺主导管结扎后萎缩腮腺内肌上皮细胞（MEC）的转归规律。方法 通过结扎大鼠右侧腮腺主导管诱导腺体萎缩,采用苏木精-伊红（HE）染色观察正常腮腺及导管结扎后1、3、5、7、14、21、30、60、 100、150 d萎缩性腮腺的组织学变化,并采用免疫组织化学染色方法定量分析MEC在腮腺萎缩不同时间点的数量及分 布情况。结果 组织学观察发现：腮腺主导管结扎5 d后,大部分腺泡细胞出现凋亡丧失,在腺体萎缩过程中形成 大量导管样结构,间质逐渐纤维化并伴有炎性细胞浸润。免疫组织化学染色定量分析结果显示：导管结扎后5 d 内,MEC数量快速增加,随后其数量增长缓慢,维持在一定的范围内,100 d时MEC数量达到峰值,其后快速减少; 在腮腺萎缩早期,MEC位于腺泡及闰管表面,呈星形或梭形,随着腺体的萎缩,最终呈梭形分布于导管样结构的外层。结论 在导管结扎后5 d内,MEC出现反应性大量增殖,随后数量增长缓慢,维持在一定的范围内;随着腮腺的逐渐萎缩,100 d后MEC数量快速减少。     

细胞外基质金属蛋白酶诱导因子表达与唾液腺肿瘤侵袭性的关系

目的:检测唾液腺肿瘤组织中细胞外基质金属蛋白酶诱导因子(EMMPRIN)的表达和定位,探讨其与唾液腺肿瘤侵袭性生物学行为的关系。方法:采用免疫组织化学方法,检测9例唾液腺正常组织、28例多形性腺瘤(PA)、25例黏液表皮样癌(MC)、33例腺样囊性癌(ACC)中EMMPRIN的表达和定位。采用SPSS10.0软件包进行多个样本率间的χ2检验或确切概率分析。结果:EMMPRIN在正常唾液腺组织、多形性腺瘤、黏液表皮样癌、腺样囊性癌的表达阳性率分别为11.11%、53.71%、84.00%和90.91%(P<0.05)。在正常唾液腺组织的表达主要见于导管上皮细胞的细胞膜;EMMPRIN蛋白在多形性腺瘤、黏液表皮样癌、腺样囊性癌的表达见于肿瘤细胞的胞膜或胞质。腺样囊性癌嗜神经现象组中,EMMPRIN表达的阳性率高于无嗜神经现象组,但差异无统计学意义。结论:EMMPRIN的表达与唾液腺肿瘤侵袭性生物学行为密切相关。     

Electron transfer kinetics and morphology of cytochrome c at the biomimetic phospholipid layers

Electrochemistry of cytochrome c (cyt c) at biomimetic phospholipid layers was studied in a phosphate buffer solution, which were formed with dilauroyl phosphatidic acid (DLPA, C12:0), dipalmitoyl phosphatidic acid (DPPA, C16:0), distearoyl phosphatidic acid (DSPA, C18:0), and palmitoyl–oleoyl phosphatidic acid (POPA, C16:0–18:1). The lipid-layers formed firstly at the air/water interface were immediately transferred onto the electrode surface using the Langmuir–Blodgett (LB) technique. The electrochemical properties of cyt c at the lipid covered electrodes depended on the orientation, number of layers of phospholipids, tail (or head) group down, and vice versa. Atomic force microscopy (AFM) images of cyt c adsorbed on the POPA monolayer (showing the head group diameter of POPA to be ca. 0.7 nm) formed on highly oriented pyrolytic graphite (HOPG) displayed uniform surface morphology of lipid layer and clumps of aggregated cyt c molecules with a minimum size corresponding to four cyt c molecules. The heterogeneous electron transfer rate constants, k⁰ values, of cyt c were determined to be 1.02 × 10⁻³, 0.98 × 10⁻³, and 0.67 × 10⁻³ cm/s for the lipid monolayer in the tail down orientation (X-type) of POPA, DLPA, and DPPA, and 0.67 × 10⁻³ and 0.50 × 10⁻³ cm/s for the head down orientation (Z-type) of POPA and DLPA monolayers, respectively.