Antigens, Neoplasm ; genetics ; Apoprotein(a) ; genetics ; Carrier Proteins ; genetics ; Endometriosis ; metabolism ; pathology ; Endometrium ; metabolism ; pathology ; Female ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; Membrane Proteins ; genetics ; Mutation, Missense ; genetics ; Nuclear Proteins ; genetics ; Ovarian Neoplasms ; metabolism ; pathology ; Proprotein Convertase 5 ; genetics ; Salivary Cystatins ; genetics ; Ubiquitin-Protein Ligases ; genetics ; Whole Exome Sequencing

Inflammatory bowel disease (IBD) is an intestinal immune-dysfunctional disease worldwide whose prevalence increasing in Asia including China. It is a chronic disease of the gastrointestinal tract with unknown cause. Exosomes are small vesicles in various body fluids. They have diameters of 40-120 nm, and one of their functions is long-distance transfer of various substances. In this study, we investigated the contents of salivary exosomes in patients with IBD and in healthy controls to explore a new biomarker in patients with IBD. In this study, whole saliva was obtained from patients with IBD (ulcerative colitis (UC), n = 37; Crohn's disease (CD), n = 11) and apparently healthy individuals (HC, n = 10). Salivary exosomes were extracted from samples, and the proteins within the exosomes were identified by liquid chromatograph-mass spectrometer (LC-MS/MS). The results showed that more than 2000 proteins were detected in salivary exosomes from patients with IBD. Through gene ontology analysis, we found that proteasome subunit alpha type 7 (PSMA7) showed especially marked differences between patients with IBD and the healthy controls, in that its expression level was much higher in the CD and UC groups. This exosomal protein is related to proteasome activity and inflammatory responses. So we conclude that in this research, salivary exosomal PSMA7 was present at high levels in salivary exosomes from subjects with IBD. It can be a very promising biomarker to release the patients from the pain of colonoscopy.
Animals ; Biomarkers ; metabolism ; Female ; Humans ; Inflammatory Bowel Diseases ; metabolism ; Male ; Proteasome Endopeptidase Complex ; metabolism ; Salivary Proteins and Peptides ; metabolism

OBJECTIVETo investigate the correlation between nurse job burnout and salivary lysozyme activity.

METHODSThe saliva samples of 131 subjects were collected at four time points for two consecutive days with saliva collection tubes. The acquisition time points were 8:00 (baseline concentration), 10:00 (morning), 15:30 (afternoon), and 17:30 (recovery period). At the same time every subjects completed the job burnout questionnaire to investigate their general demographic characteristics and job burnout level. The salivary lysozyme concentration was measured with ELISA. The data were analyzed by partial correlation analysis and multiple stepwise regression analysis.

RESULTSThere were significant differences in the salivary lysozyme activity between subjects with different ages, working years, and education levels. The work period vitality and the average energy of ≤ 30 age group were higher than other two groups and the recovery energy was higher than >35 age group. Working period vitality, the average energy of group >15 years were less than ≤ 10 years group. The work period energy and the average energy of university (college) and above group were lower than high school (secondary) and the following group. Job burnout and its three dimensions had a significant negative correlation with salivary lysozyme concentration (P < 0.01). Depersonalization and emotional exhaustion were the negative impact factors for salivary lysozyme activity at baseline. Emotional exhaustion and personal fulfillment were the negative impact factors for salivary lysozyme activity during the working period. Personal fulfillment was the negative factor for salivary lysozyme activity during the recovery period and the average salivary lysozyme activity.

CONCLUSIONSalivary lysozyme activity is sensitive for nurse job burnout, so it can be used as an objective evaluation index of job burnout.

Burnout, Professional ; epidemiology ; psychology ; Emotions ; Fatigue ; Humans ; Muramidase ; analysis ; Nurses ; psychology ; Occupational Diseases ; epidemiology ; psychology ; Regression Analysis ; Salivary Proteins and Peptides ; analysis ; Surveys and Questionnaires

Lysophosphatidic acid (LPA) is a bioactive lysophospholipid involved in numerous physiological responses. However, the expression of LPA receptors and the role of the Hippo signaling pathway in epithelial cells have remained elusive. In this experiment, we studied the functional expression of LPA receptors and the associated signaling pathway using reverse transcriptase-PCR, microspectrofluorimetry, western blotting and immunocytochemistry in salivary gland epithelial cells. We found that LPA receptors are functionally expressed and involved in activating the Hippo pathway mediated by YAP/TAZ through Lats/Mob1 and RhoA/ROCK. Upregulation of YAP/TAZ-dependent target genes, including CTGF, ANKRD1 and CYR61, has also been observed in LPA-treated cells. In addition, based on data suggesting that tumor necrosis factor (TNF)-alpha induces cell apoptosis, LPA upregulates TNF-induced caspase-3 and cleaved Poly(ADP-ribose)polymerase (PARP). However, small interfering RNA treatment to Yes-associated protein (YAP) or transcriptional co-activator with a PDZ-binding motif (TAZ) significantly decreased TNF-alpha- and LPA-induced apoptosis, suggesting that YAP and TAZ modulate the apoptotic pathway in salivary epithelial cells.
Adaptor Proteins, Signal Transducing/genetics/metabolism ; *Apoptosis ; Cell Line ; Epithelial Cells/*cytology/metabolism ; Gene Expression Regulation ; Humans ; Intracellular Signaling Peptides and Proteins/genetics/metabolism ; Lysophospholipids/*metabolism ; Phosphoproteins/genetics/metabolism ; Protein-Serine-Threonine Kinases/*metabolism ; RNA Interference ; RNA, Small Interfering/genetics ; Receptors, Lysophosphatidic Acid/genetics/*metabolism ; Salivary Glands/*cytology/metabolism ; *Signal Transduction ; Tumor Necrosis Factor-alpha/metabolism ; rho-Associated Kinases/metabolism ; rhoA GTP-Binding Protein/metabolism

Botulinum toxin A (BTXA) has been used in several clinical trials to treat excessive glandular secretion; however, the precise mechanism of its action on the secretory function of salivary gland has not been fully elucidated. In this study, we aimed to investigate the effect of BTXA on secretion of submandibular gland in rabbits and to identify its mechanism of action on the secretory function of salivary gland. At 12 weeks after injection with 5 units of BTXA, we found a significant decrease in the saliva flow from submandibular glands, while the salivary amylase concentration increased. Morphological analysis revealed reduction in the size of acinar cells with intracellular accumulation of secretory granules that coalesced to form a large ovoid structure. Expression of M3-muscarinic acetylcholine receptor (M3 receptor) and aquaporin-5 (AQP5) mRNA decreased after BTXA treatment, and distribution of AQP5 in the apical membrane was reduced at 1, 2 and 4 weeks after BTXA injection. Furthermore, BTXA injection was found to induce apoptosis of acini. These results indicate that BTXA decreases the fluid secretion of submandibular glands and increases the concentration of amylase in saliva. Decreased expression of M3 receptor and AQP5, inhibition of AQP5 translocation, and cell apoptosis might involve in BTXA-reduced fluid secretion of submandibular glands.
Amylases ; drug effects ; Animals ; Apoptosis ; drug effects ; Aquaporin 5 ; antagonists & inhibitors ; Botulinum Toxins, Type A ; pharmacology ; Cell Membrane ; drug effects ; In Situ Nick-End Labeling ; Male ; Microscopy, Electron, Transmission ; Neuromuscular Agents ; pharmacology ; Organ Size ; Rabbits ; Random Allocation ; Receptor, Muscarinic M3 ; antagonists & inhibitors ; Saliva ; drug effects ; secretion ; Salivary Proteins and Peptides ; drug effects ; Salivation ; drug effects ; Secretory Rate ; drug effects ; Secretory Vesicles ; drug effects ; Submandibular Gland ; drug effects ; pathology ; secretion ; Time Factors

There is a need recognized by the National Institute of Dental & Craniofacial Research and the National Cancer Institute to advance basic, translational and clinical saliva research. The goal of the Salivaomics Knowledge Base (SKB) is to create a data management system and web resource constructed to support human salivaomics research. To maximize the utility of the SKB for retrieval,integration and analysis of data, we have developed the Saliva Ontology and SDxMart. This article reviews the informatics advances in saliva diagnostics made possible by the Saliva Ontology and SDxMart.
Biomarkers ; chemistry ; Computational Biology ; methods ; Databases, Protein ; Genomics ; methods ; Humans ; Metabolomics ; methods ; Proteomics ; methods ; Saliva ; chemistry ; Salivary Proteins and Peptides ; chemistry ; classification ; physiology

OBJECTIVETo study the role of Slit-Robo signaling in the proliferation of human mucoepidermoid carcinoma Mc3 cells.

METHODSWe measured the expressions of Slit2 and Robo1 proteins in human mucoepidermoid carcinoma Mc3 cells using immunohistochemistry and flow cytometry. To test the role of Slit-Robo signaling in the proliferation of the cells, we treated the cells with the monoclonal antibody R5 against Robo1 receptor extracellular domain and observed the changes in the cell proliferation by cell counting and colonogenic assay; the expression of proliferating cell nuclear antigen (PCNA) in the cells following the treatment was measured with flow cytometry.

RESULTSTreatment of Mc3 cells with the monoclonal antibody R5 caused significantly suppressed cell growth and proliferation and obviously lowered the expression of PCNA.

CONCLUSIONSlit-Robo signaling can suppress the proliferation of Mc3 cells possibly by up-regulating the expression of PCNA.

Carcinoma, Mucoepidermoid ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Intercellular Signaling Peptides and Proteins ; metabolism ; Nerve Tissue Proteins ; metabolism ; Proliferating Cell Nuclear Antigen ; metabolism ; Receptors, Immunologic ; metabolism ; Salivary Gland Neoplasms ; pathology ; Signal Transduction

OBJECTIVETo study the effect of reserpine (RSP) for changing salivary protein secretion in Pi-deficient rats and to explore its possible mechanism.

METHODSTwenty rats allocated in the RSP group were given subcutaneous injection of RSP [0.4 mg/(kg x d)] for 9 successive days, while the other 20 rats in the control group were injected with same volume of saline instead. On the 10th day, ten rats randomly selected from each group were subjected for extracting saliva to detect salivary amylase activity (sAA) before and after an acid stimulation; and drawing blood from the orbital vein to measure the contents of vasoactive intestinal peptide (VIP) and cyclic adenosine monophosphate (cAMP). Then they were sacrificed and their parotids were taken out for pathological examination with HE staining, as well as for VIP and cAMP measuring, and zymogen granules counting under a transmission electron microscope. The remainder animals were stopped injecting and normally fed to 40 days, then subjected to be detected as above-mentioned.

RESULTSFood intake and body weight reduction were more significantly in the RSP group than in the control group. On the 10th day, the ratio of sAA before/after stimulation in the RSP group was 0.39 +/- 0.18, significantly lower than that in the control group (0.80 +/- 0.21, P < 0.01), but it was restored rapidly, reaching the normal range on the 25th day, on the 40th day, it became significantly different to the level on the 10th day (P < 0.05) and approached the level in the control group (P > 0.05). No significant pathological change of parotid was found in both groups; but the number of zymogen granules in the RSP group was remarkably more than that in the control group (41.4 +/- 4.9 vs 34.6 +/- 5.2, P < 0.01). Serum level of VIP in the RSP group was significantly less while that of cAMP was higher than that in the control group (22.5 +/- 13.1 mg/L vs 38.5 +/- 14.1 mg/L, and 125.8 +/- 15.5 micromol/L vs 105.3 +/- 16.7 micromol/L, both P < 0.05), but no inter-group difference was found in parotid tissue contents of both VIP and cAMP. All the indices detected became equivalent in the two groups on the 40th day.

CONCLUSIONThe reduction of salivary protein in Pi-deficient rats induced by RSP may be related to the regulatory pathway of VIP and cAMP.

Animals ; Cyclic AMP ; blood ; Male ; Medicine, Chinese Traditional ; Rats ; Rats, Sprague-Dawley ; Reserpine ; adverse effects ; pharmacology ; Salivary Proteins and Peptides ; metabolism ; Salivation ; drug effects ; Vasoactive Intestinal Peptide ; blood

Salivary glands provide saliva to maintain oral health, and a loss of salivary gland function substantially decreases quality-of-life. Understanding the biological mechanisms that generate salivary glands during embryonic development may identify novel ways to regenerate function or design artificial salivary glands. This review article summarizes current research on the process of branching morphogenesis of salivary glands, which creates gland structure during development. We highlight exciting new advances and opportunities in studies of cell-cell interactions, mechanical forces, growth factors, and gene expression patterns to improve our understanding of this important process.
Animals ; Cell Communication ; physiology ; Embryonic Development ; physiology ; Epithelium ; embryology ; Extracellular Matrix ; physiology ; Humans ; Intercellular Signaling Peptides and Proteins ; physiology ; Morphogenesis ; physiology ; Salivary Glands ; embryology

BACKGROUNDA20, also known as tumor necrosis factor alpha induced protein 3 (TNFaip3), is a cytoplasmic zinc finger protein that inhibits nuclear factor kappa-B (NF-kappaB) activity and prevents tumor necrosis factor (TNF)-mediated programmed cell death. NF-kappaB is a transcription factor that regulates expression of genes involved in cell proliferation, cell survival and anti-apoptosis. Several studies have implicated that the NF-kappaB signal pathway is associated with angiogenesis and clinico-pathological process of adenoid cystic carcinoma (ACC) of the salivary glands.

METHODSThe ability of overexpression of A20 to influence the biological behavior and invasion of ACC cells was examined. The cells were stably transfected with full-length A20 cDNA. Stable gene transfer was verified by realtime-polymerase chain reaction (PCR) and Western blot analysis. The change of cell biological behavior was examined by methyl thiazolyl tetrazolium (MTT) and NF-kappaB luciferase reporter assay and the invasion of the cells was examined by a Matrigel invasion chamber.

RESULTSpEGPFN3-A20 gene was stably transferred into ACC-2 cells and overexpressed. When cells were treated with TNFalpha, the NF-kappaB activity of ACC-2-A20 cells could be down-regulated about 46.32% in contrast to ACC-2-GFP cells (P < 0.05). A20 potently inhibited growth of A20 transfectant ACC-2-A20 compared with control vector transfected groups and the ACC-2 empty control group (P < 0.05). The ACC-2-A20 cells showed significantly reduced ability to invade through Matrigei-coated filters compared to ACC-2-GFP and ACC-2 cells. The inhibition rate was up to 71.05% (P < 0.05).

CONCLUSIONSA20 gene transfer is associated with decreased tumor invasion, in part via the down-regulation of NF-kappaB expression, providing evidence for a potential application of A20 in designing a treatment modality for salivary gland cancers such as ACC.

Carcinoma, Adenoid Cystic ; pathology ; therapy ; Cell Line, Tumor ; DNA-Binding Proteins ; Genetic Therapy ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; NF-kappa B ; antagonists & inhibitors ; Neoplasm Invasiveness ; Nuclear Proteins ; genetics ; Salivary Gland Neoplasms ; pathology ; therapy ; Transfection ; Tumor Necrosis Factor alpha-Induced Protein 3