A study of lectin bindings to the fetal membranes.

Human tissues contain carbohydrates for a main component, functioning as a source and reservoir of energy, connective and supporting element, recognition site and related tasks. Our main interest is to reveal the synthesis and distribution of carbohydrate elements in human fetal membranes. The aim of our work was to clarify, which kinds of elements containing carbohydrates, existed in the fetal membranes. Therefore we applied a lectin-binding study using the following FITC labelled lectins: ConA, WGA, PNA, LCA, RCA. This lead to the result, that ConA, LCA, WGA and RCA produced a positive reaction in the amnion epithelium, which was negative when using PNA. The basement membrane I showed an intense fluorescence when we used ConA, LCA and WGA, using RCA it was weaker and using PNA fluorescence was nearly missing. The examination of the amniotic fibroblast and intercellular substance showed a positive reaction with all lectins, but the intercellular substance lead to weaker fluorescence. The chorionic fibroblasts, intercellular substance and basement membrane II produced fluorescence using ConA, LCA, WGA and PNA, but no reaction could be examined, when using RCA. The trophoblastic cells did not react with LCA and RCA. The intercellular substance reacted positively with all lectins.     

Metastatic uterine leiomyosarcoma to the submandibular gland

Metastasis of uterine leiomyosarcoma to the salivary glands is exceedingly rare. In this report, we present the clinicopathologic features of one such case. The patient was diagnosed with uterine leiomyosarcoma at age 58 years and was treated by hysterectomy only. The tumor was confined to the uterine body. The patient remained free of disease for 4 years and then developed a left submandibular mass. Imaging studies also showed lung involvement but no tumor in the abdomen or pelvis. The submandibular mass was resected; it had histologic and immunohistochemical features similar to those seen in the previous uterine leiomyosarcoma. After 7 months of follow-up, the patient is alive with disease, with progression of disease in the lungs despite receiving chemotherapy.     

胎儿颌下腺内神经肽免疫组织化学研究

目的 :对胎儿颌下腺内神经肽进行定性和定位研究 ,以探讨其功能属性。方法 :采用免疫组织化学ABC方法。结果 :第 16～ 18周 ,胎儿颌下腺呈P物质 (SP)、血管活性肠肽 (VIP)及神经肽Y (NPY)免疫反应性 ,为弱阳性～中等阳性 ,主要分布于导管上皮胞质内 ,细胞核为阴性。至 2 2～ 2 4周左右 ,SP和VIP免疫反应强度达到强阳性反应 ,随后反应强度减弱。NPY无明显增龄变化。结论 :(1)人胎颌下腺内含有SP、VIP及NPY等神经肽类物质 ;(2 )这些神经肽共存于颌下腺导管上皮细胞内 ,上述结果为深入研究颌下腺分泌功能属性提供了形态学资料     

Subcellular localization of epidermal growth factor receptor in human submandibular gland

The subcellular distribution of the epidermal growth factor receptor (EGFr) was demonstrated in the normal human submandibular gland by means of immunogold cytochemistry. EGFr labelling appeared in both acinar and ductal cells, where strong immunoreactivity was associated with a tubulovesicular system near the basolateral surfaces. In addition, groups of reactive vesicles were highlighted among secretory granules of both serous and mucous cells and at the apex of ductal cells. Basolateral vesicles were interpreted as being a result of EGFr internalization after activation by an exogenous ligand, although the functional meaning of those located apically remains unclear.     

Reaction of the rat submandibular salivary gland to injury to the contralateral gland

After burns of resection of the submandibular salivary gland the intact contralateral gland in rats responds by increased proliferative activity. The number of mitoses reached a maximum 72 h after injury in the case of burns and 48 h after resection. Burns of the salivary gland cause lasting but weak compensatory hypertrophy of the contralateral gland. Hypertrophy of the gland is accompanied by an increase in size of the cells and nuclei, the area of which rises by 10 and 17% respectively. Resection of the salivary gland causes an increase in weight of the intact gland only in the early period of observation; by the 30th and 45th days after the operation the weight of the experimental glands was not significantly different from the control. Differences in compensatory growth of the intact glands observed after two types of injury of the contralateral gland evidently depend on the quantity of tissue breakdown products and the duration of their presence in the body.Department of Biology and General Genetics, Moscow Medical Stomatological Institute. (Presented by Academician of the Academy of Medical Sciences of the USSR A. P. Avtsyn.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 82, No. 9, pp. 1108–1110, September, 1976.     

Nodular fasciitis in the submandibular gland

Nodular fasciitis (NF) is a benign, proliferative lesion of myofibroblasts. The most common site of the lesion is in the upper extremities or trunk. NF in the head and neck is next in frequency and is the most common site in infants and children. In adults, NF in the submandibular region is very rare. We report a case of a 53-year-old man with a submandibular mass, rapidly growing for 10 days. Cytologic findings revealed a few sheets or clusters of small, uniform spindle cells. The uniform spindle cells had centrally located round to ovoid nucleus, but there were no nuclear atypia and atypical mitosis. There were focally loose stroma but we could not find chondroid or myxoid components. A partial parotidectomy was performed. We describe a case of NF in the submandibular region, adjacent to the parotid gland and review the clinical, cytologic, histologic, and immunohistochemical features of NF.     

Histopathological study of the human submandibular gland in graft versus host disease

Major salivary gland dysfunction and severe xerostomia is one of the manifestations of graft versus host disease (GVHD). The histopathological evaluation of the major salivary gland in patients with GVHD has never been reported. The pathological findings of the submandibular glands in a GVHD patient who succumbed to the disease are described. Lymphocytic infiltration, parenchymal destruction, and fibrosis were observed, which may provide the pathophysiological mechanism for the xerostomia and hyposalivation observed in GVHD.     

下颌下腺移位预防放疗口干燥症的应用解剖

目的　为下颌下腺移位于颏下间隙预防头颈部放疗后口干燥症提供解剖学基础。 方法　30侧经颈外动脉灌注红色乳胶的正常成人头颈部标本,在2.0~3.0倍的便携式手术显微镜下对下颌下腺及血管进行显微解剖,观测下颌下腺腺体的体积、下颌下腺导管的长度及腺体周围动静脉腺支血管的走行、分布、毗邻关系等显微解剖特点。 结果　下颌下腺以下颌舌骨肌后缘为界分为较大的浅部和小的深部。浅部体积为（1.7±0.5）cm3;深部为下颌下腺的延长部分,体积为（1.0±0.8）cm3。下颌下腺导管由下颌下腺浅部的深面发出,长约5cm,开口于舌系带旁的舌下肉阜。 结论　下颌下腺移位于颏下间隙预防头颈部放疗后口干燥症的手术具有可行性。     

Phenotypic expression of human epidermal growth factor in foetal submandibular gland and pleomorphic adenoma of salivary gland

Summary The phenotypic expression of the human epidermal growth factor (EGF) was investigated immunohistochemically in human foetal submandibular glands from the 5th to 10th month of gestation, adult normal submandibular glands and 48 cases of pleomorphic adenomas. In foetal submandibular glands, both the terminal buds and primary ducts at the intermediate stage of gestation were positive for EGF, and in particular, the outer layer cells of primary ducts showed strong EGF-immunoreactivity. EGF-positive cells decreased as the gestational stage advanced and only ductal cells were weakly positive for EGF at the terminal stage of gestation. In the adult normal submandibular gland, weak immunoreactivity for EGF was restricted to ductal cells. However, 41 (86%) of the 48 pleomorphic adenomas had EGF-positive cells which were distributed among the ductal, chondroid and myxoid portion. No EGF-immunoreactivity was detected in the solid portion of pleomorphic adenomas. These results suggest that EGF may play an important role in the growth and differentiation of foetal cells as well as the proliferation of tumour cells in pleomorphic adenomas.     

Detection of human herpesvirus 6 and human herpesvirus 7 in the submandibular gland, parotid gland, and lip salivary gland by PCR.

In order to define the major sites of persistence of human herpesvirus 6 (HHV-6) and HHV-7, PCR with DNAs from more than 100 specimens of 3 different salivary glands was performed. HHV-6 DNA was detected in 52 (88.1%) of 59 submandibular gland, 17 (63.0%) of 27 parotid gland, and 9 (52.9%) of 17 lip salivary gland specimens. On the other hand, HHV-7 DNA was detected in 59 (100%) of 59 submandibular gland, 23 (85.2%) of 27 parotid gland, and 10 (58.8%) of 17 lip salivary gland specimens. These findings demonstrate that salivary glands are a site of persistent infection of both HHV-6 and HHV-7 and that among the three types of salivary gland examined, the submandibular gland is the primary one in which these herpesviruses, especially HHV-7, persist.     

Distribution of calcitonin gene-related peptide immunoreactive nerve fibers in the human submandibular gland

The indirect immunofluorescence technique was used to study the distribution of calcitonin gene-related peptide (CGRP) in human submandibular gland. A relatively low number of thin varicose fibers with intense immunofluorescence for CGRP was seen in samples from seven glands. These CGRP-immunoreactive (CGRP-IR) nerve fibers were mainly seen around or in close contact with intra- and interlobular blood vessels. Some CGRP-IR nerve fibers were also found in association with intra- and interlobular salivary ducts and a few around the submandibular acini. By visual estimation there was no difference in the density of CGRP-IR nerve fibers between specimens of recurrent duct obstruction and laryngeal carcinoma. The present results show that the distribution of CGRP-IR nerve fibers in the stroma and in the glandular secretory elements of the human submandibular gland is quite similar to that seen in the rat and the ferret, which have been reported earlier. Furthermore, the regional distribution of CGRP-IR fibers in the human submandibular gland suggests that CGRP has a physiological role in the regulation of salivary gland function in human salivary glands, e.g. blood flow and secretion.     

Enhanced immunocytochemical expression of antioxidant enzymes in rat submandibular gland after normobaric oxygenation

In order to clarify the role of antioxidant enzymes in the male rat submandibular gland against short-term normobaric oxygenation, we performed immunocytochemical staining of manganese-containing superoxide dismutase (Mn-SOD), copper- and zinc-containing SOD (Cu/Zn-SOD), catalase (CAT), glutathione peroxidase, and glutathione S-transferases (GST alpha, GST mu, and GST pi) between days 1 and 7 after normobaric oxygenation. Ultrastructural alterations and immunoreactivities for malondialdehyde (MDA), a lipid peroxidation-related molecule, of the acinar and ductal cells after the oxygenation were also investigated. Immunoreactivity for MDA was exhibited in the acinar cells throughout the experiment. On the other hand, immunoreactivity for the SODs, CAT, and GSTs was not altered, when compared to that of controls, but was significantly elevated in the granular, striated, and excretory ductal cells. Since an increase of lipid peroxidation as indicated by enhanced immunoreactivity for MDA was detected in the acinar and intercalated ductal cells, the results indicate that the enhanced antioxidant enzymes in the granular, striated, and excretory ductal cells play a crucial role in the self-defense system of the male rat submandibular gland against normobaric oxygenation.     

Spontaneous salivation in the rabbit submandibular gland

1. Salivation has been studied in the submandibular gland of the rabbit. A very slow spontaneous salivation took place when all possibility of nerve influence had been excluded. Salivation was not due to ultrafiltration.2. The ;spontaneous' saliva had a mean K concentration of 148 mM and Na concentration of 46 mM. With increasing salivation rate produced by parasympathetic nerve stimulation, K concentration fell to a plateau level of about 30 mM whilst Na concentration fell rapidly to reach the low value of 3 mM, then began to rise again at the higher flow rates.3. Ligation of the submandibular duct produced a reversal of the ion concentrations in spontaneous saliva. By 4 days K concentration was lower and that of Na higher than control values until by 2 weeks the effect was maximal with mean concentrations of 25 mM for K and 153 mM for Na.4. Ouabain increased the spontaneous salivation rate and ethacrynic acid slowed or prevented it altogether. On the basis of the known sites of action of these drugs it is postulated that two pumps are involved in the regulation of spontaneous salivation. There appears to be basal activity of an acinar mechanism pumping NaCl into the lumen, taking water with it. This pump is activated directly or indirectly by the intracellular Na concentration which itself is controlled by an Na-K exchange pump.5. Excitation of the sympathetic trunk produced a small, though definite, increase in salivation rate. There was evidence that myoepithelial cells might also be involved in the sympathetic response and that they were activated by alpha receptor stimulation. Salivation evoked by sympathetic nerve stimulation would seem to be a response to beta receptor stimulation, but the possibility that activation of both alpha and beta receptors was required could not be excluded entirely.     

Regeneration of acini in submandibular gland autografts

Regeneration of submandibular gland (SMG) secretory parenchyma is remarkably impaired in salivary gland diseases and under experimental conditions such as in tissue culture and after isografting. In our study acinar regeneration was found to depend on the site where the SMG tissue was implanted. Implantation of several 2-3 mm3 fragments of SMG subcutaneously in the back of the same donor adult male rat resulted in initial necrosis and mononuclear cell infiltration of the autograft. Then there was epithelial proliferation with the appearance within 28 days of lobules which contained numerous duct-like structures and only a few or no acini. In contrast, implanting SMG fragments in the anatomical bed of the donor gland resulted in the appearance of a more differentiated autograft. Although the initial tissue changes were similar to those seen in the autografts in the subcutaneous tissues of the back, the SMG autograft in the neck also contained numerous acini by 42 and 56 days after implantation. These data support the view that the implantation site influences the course of cytodifferentiation in SMG autografts.     

Expression and localization of chromogranin A gene and protein in human submandibular gland

Human saliva chromogranin A (CgA) is clinically promising as a psychological stress marker. However, expression of CgA is poorly understood in humans, although salivary gland localization of CgA in other mammals, such as rodents and horses, has been demonstrated. In the present study, we investigated the expression and localization of CgA in the human submandibular gland (HSG) using various methods. CgA was consistently localized in serous and ductal cells in HSG, as detected by immunohistochemistry and in situhybridization. Reactivity was stronger in serous cells than in ductal cells. In addition, strong immunoreactivity for CgA was observed in the saliva matrix of ductal cavities. Western blotting gave one significant immunoreactive band of 68 kDa in the adrenal gland, HSG and saliva. Finally, CgA was detected in secretory granules of serous and ductal cells by immunoelectron microscopy. In conclusion, CgA in humans is produced by HSG and secreted into saliva.     

Differentiation of primary human submandibular gland cells cultured on basement membrane extract

There is no effective treatment for the loss of functional salivary tissue after irradiation for head and neck cancer or the autoimmune disease Sj?gren's syndrome. One possible approach is the regeneration of salivary glands from stem cells. The present study aimed to investigate whether small pieces of human submandiblar gland tissue contain elements necessary for the reconstruction of salivary rudiments in vitro via acinar and ductal cell differentiation. Primary submandibular gland (primary total human salivary gland; PTHSG) cells were isolated from human tissue and cultured in vitro using a new method in which single cells form an expanding epithelial monolayer on plastic substrates. Differentiation, morphology, number, and organization of these cells were then followed on basement membrane extract (BME) using RNA quantitation (amylase, claudin-1 (CLN1), CLN3, kallikrein, vimentin), immunohistochemistry (amylase and occludin), viability assay, and videomicroscopy. On the surface of BME, PTHSG cells formed acinotubular structures within 24 h, did not proliferate, and stained for amylase. In cultures derived from half of the donors, the acinar markers amylase and CLN3 were upregulated. The PTHSG culture model suggests that human salivary gland may be capable of regeneration via reorganization and differentiation and that basement membrane components play a crucial role in the morphological and functional differentiation of salivary cells.     

A proteomics approach to characterizing human submandibular gland cell lines by fluorescent two-dimensional differential in-gel electrophoresis

The human salivary glands have a variety of histologic features such as intercalated duct cells, myoepithelial cells and acinar cells. A neoplastic human salivary intercalated duct cell line (HSG) and its derivatives, HSG with a myoepithelial cell phenotype (HSG-AZA1) and HSG with an acinar cell phenotype (HSG-AZA3) induced by 5-aza-2'-dC treatment of HSG cells, have been reported. To identify characterization of intercalated duct cells, myoepithelial cells and acinar cells in the salivary gland, we selected HSG, HSG-AZA1 and HSG-AZA3 cell lines to perform two-dimensional electrophoresis analysis. We used a fluorescent two-dimensional differential in-gel electrophoresis (2-D-DIGE) for comparative proteomics, which improved the reproducibility and reliability of differential protein expression analysis between the samples. Furthermore, matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) peptide mass fingerprinting (PMF) was used to identify the proteins. These methods were combined to approach the protein profiles associated with characterization between HSG, HSG-AZA1 and HSG-AZA3 cells. Using these strategies, we identified seven HSG associated proteins, such as actin-beta, hydrocephalus inducing protein, L-plastin, KIAA0657 protein, septin 6 isoform A, lamin A/C isoform 2 and superoxide dismutase 2, three HSG-AZA1 associated proteins such as ubiquitin carboxyl-terminal esterase L1, myosin light chain 2 and muscle creatine kinase, and two HSG-AZA3 associated proteins, microtubule-associated protein 6 and Annexin A3. These results suggest that the proteins are associated with characterization of the salivary gland.     

胎儿下颌下腺内瘦素和瘦素受体的表达及定位

目的：观察胎儿下颌下腺内瘦素和瘦素受体的表达、分布及发育规律。方法：HE染色法和免疫组织化学SABC法。结果：胎儿下颌下腺内纹状管和小叶问导管上皮细胞呈瘦素及瘦素受体免疫阳性反应,免疫反应产物分布于导管上皮细胞胞质内,细胞核星阴性反应。在胚胎发育的不同阶段,免疫反应强度亦不等。结论：瘦素和瘦素受体表达于胎儿下颌下腺内,可能参与调节胎儿下颌下腺和胃肠的发育及功能活动。