This study investigated the therapeutic effect of Shegan Mahuang Decoction(SGMHD) on cold-induced asthma in rats and explored its underlying mechanism. Seventy-two healthy male SD rats of specific pathogen free(SPF) grade were randomly divided into a blank group, a model group, a positive control group(dexamethasone, 0.4 mg·kg~(-1)), and low-, medium-, and high-dose SGMHD groups(3.2, 6.4, and 12.8 g·kg~(-1)). The blank group received saline, while the other groups were sensitized by intraperitoneal injection of ovalbumin(OVA) solution. Subsequently, the rats were placed in a cold chamber adjustable to 0-2 ℃, and OVA solution was ultrasonically nebulized to induce cold-induced asthma in rats. After three weeks of treatment, the general behaviors of rats were observed. Hematoxylin-eosin(HE) staining was used to evaluate pathological changes in lung tissues, periodic acid-Schiff(PAS) staining assessed mucin changes, and Masson staining was performed to examine collagen deposition. Enzyme-linked immunosorbent assay(ELISA) was used to measure the levels of the inflammatory factors interleukin-4(IL-4) and vascular endothelial growth factor(VEGF) in serum and bronchoalveolar lavage fluid(BALF). Real-time quantitative polymerase chain reaction(RT-PCR) was employed to assess the mRNA expression levels of transient receptor potential vanilloid subfamily member 1(TRPV1), nuclear respiratory factor 1(NRF-1), and mitochondrial transcription factor A(mtTFA) in lung tissues. Western blot was used to measure the protein expression levels of TRPV1, NRF-1, and mtTFA in lung tissues. Compared with the blank group, the model group exhibited signs of rapid respiration, increased frequency of defecation with looser stools, and disheveled and dull fur. Pathological results showed significant infiltration of inflammatory cells in lung tissues, narrowing of bronchial lumens, increased mucin secretion, and enhanced collagen deposition in the model group. Additionally, the levels of IL-4 and VEGF in serum and BALF were significantly elevated, and the mRNA and protein expression levels of TRPV1, NRF-1, and mtTFA in lung tissues were significantly increased. Compared with the model group, SGMHD improved the behaviors of rats, alleviated pathological changes in lung tissues, mucin production, and collagen deposition, significantly decreased the levels of IL-4 and VEGF in serum and BALF, and reduced the mRNA expression levels of TRPV1, NRF-1, and mtTFA in lung tissues, with the medium-dose SGMHD group showing the most significant effect. Moreover, the protein expression levels of TRPV1, NRF-1, and mtTFA in lung tissues were also reduced, with the medium-dose SGMHD group exhibiting the most significant effect. In conclusion, this study demonstrates that SGMHD can alleviate airway inflammation and inhibit airway remodeling in cold-induced asthma rats. These effects may be associated with the modulation of the TRPV1/NRF-1/mtTFA signaling pathway.
Rats ; Male ; Animals ; Mice ; Interleukin-4/metabolism* ; Vascular Endothelial Growth Factor A/metabolism* ; Rats, Sprague-Dawley ; Asthma/genetics* ; Lung ; Bronchoalveolar Lavage Fluid ; RNA, Messenger/metabolism* ; Collagen/metabolism* ; Mucins/therapeutic use* ; Ovalbumin ; Disease Models, Animal ; Mice, Inbred BALB C ; TRPV Cation Channels/metabolism* ; Drugs, Chinese Herbal

Mucins,a family of heavily glycosylated proteins,present mainly in epithelial cells.They function as essential barriers for epithelium and play important roles in cellular physiological processes.Aberrant expression and glycosylation of mucins in gastric epithelium occur at pathological conditions,such as Helicobacter pylori infection,chronic atrophic gastritis,intestinal metastasis,dysplasia,and gastric cancer.This review addresses the major roles played by mucins and associated O-glycan structures in normal gastric epithelium.Further,we expound the alterations of expression patterns and glycan signatures of mucins at those pathological conditions.
Gastric Mucosa/pathology* ; Glycosylation ; Helicobacter Infections/pathology* ; Helicobacter pylori/metabolism* ; Humans ; Mucins/metabolism* ; Stomach Neoplasms/pathology*

BACKGROUND AND OBJECTIVES: Insulin is a peptide hormone that regulates the metabolism of carbohydrates and fats by promoting the absorption of glucose from the blood to skeletal muscles. Insulin has been reported to be closely related to cardiovascular, respiratory, and endocrine disease. However, the effect of insulin on production of major mucins in human airway epithelial cells has not been reported. Therefore, this study investigated the relationship between high levels of insulin and mucin in human airway epithelial cells. MATERIALS AND METHODS: This study analyzed the effect of high level of insulin on MUC4, MUC5AC, and MUC5B expression using reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay in human airway epithelial cells. RESULTS: In human NCI-H292 airway epithelial cells, high level of insulin significant increased MUC4, MUC5AC, and MUC5B mRNA expression and glycoprotein production. In the primary cultures of normal nasal epithelial cells, high level of insulin also increased MUC4, MUC5AC, and MUC5B expression. CONCLUSION: These results suggest that insulin plays a role in control of mucus hypersecretion in human airway epithelial cells.
Absorption ; Carbohydrates ; Endocrine System Diseases ; Enzyme-Linked Immunosorbent Assay ; Epithelial Cells* ; Fats ; Glucose ; Glycoproteins ; Humans ; Insulin ; Metabolism ; Mucins ; Mucus ; Muscle, Skeletal ; RNA, Messenger

This study aimed at constructing a draft genome of the adult female worm Toxocara canis using next-generation sequencing (NGS) and de novo assembly, as well as to find new genes after annotation using functional genomics tools. Using an NGS machine, we produced DNA read data of T. canis. The de novo assembly of the read data was performed using SOAPdenovo. RNA read data were assembled using Trinity. Structural annotation, homology search, functional annotation, classification of protein domains, and KEGG pathway analysis were carried out. Besides them, recently developed tools such as MAKER, PASA, Evidence Modeler, and Blast2GO were used. The scaffold DNA was obtained, the N50 was 108,950 bp, and the overall length was 341,776,187 bp. The N50 of the transcriptome was 940 bp, and its length was 53,046,952 bp. The GC content of the entire genome was 39.3%. The total number of genes was 20,178, and the total number of protein sequences was 22,358. Of the 22,358 protein sequences, 4,992 were newly observed in T. canis. Following proteins previously unknown were found: E3 ubiquitin-protein ligase cbl-b and antigen T-cell receptor, zeta chain for T-cell and B-cell regulation; endoprotease bli-4 for cuticle metabolism; mucin 12Ea and polymorphic mucin variant C6/1/40r2.1 for mucin production; tropomodulin-family protein and ryanodine receptor calcium release channels for muscle movement. We were able to find new hypothetical polypeptides sequences unique to T. canis, and the findings of this study are capable of serving as a basis for extending our biological understanding of T. canis.
Adult ; B-Lymphocytes ; Base Composition ; Classification ; DNA ; Female ; Genome* ; Genomics ; Humans ; Larva Migrans, Visceral* ; Metabolism ; Mucins ; Peptides ; Protein Structure, Tertiary ; Receptors, Antigen, T-Cell ; RNA ; Ryanodine Receptor Calcium Release Channel ; T-Lymphocytes ; Toxocara canis* ; Toxocara* ; Transcriptome ; Ubiquitin-Protein Ligases

OBJECTIVETo study the clinicopathologic features of combined hepatocellular-cholangiocarcinoma (cholangiolocellular type, CLC type) with stem cell features and its relationship to hepatic progenitor cells (HPCs).

METHODSClinical and histologic features of 26 cases of combined hepatocellular-cholangiocarcinoma (CLC type) were reviewed. Histochemistry was performed to confirm the type of mucin and immunohistochemical study was carried out for hepatocytic markers (Hep Par-1 and AFP) and biliary/HPCs markers (CK7, CK9, EMA, EpCAM, NCAM, CKIT).

RESULTSThe age of patients ranged from 51 to 82 years (mean 64 years). All 26 cases contained CLC and hepatocellular carcinoma components. CLC area was composed of mixtures of small monotonous glands with abundant fibrous stroma and lymphocytic infiltrate. Tumor cells were cuboidal, smaller in size than normal hepatocytes, with basophilic cytoplasm and round nuclei. All cases, especially at the tumor boundary, showed HCC-like trabecular areas characterized by mildly atypical tumor cells with abundant eosinophilic cytoplasm and little stroma. Out of 26 cases, 21 showed definite glandular formation with mucin production, representing intrahepatic cholangiocarcinoma areas. The three distinct areas showed transitional zones merging with each other. The surrounding liver tissue showed cirrhosis and chronic hepatitis with varying degrees of fibrosis and periportal ductular reaction. Immunohistochemistry showed that biliary/HPC markers (CK7, CK9, EMA, EpCAM, NCAM and CKIT) were strongly positive in CLC area in almost all cases, similar to the staining pattern of ductular reaction. In HCC-like areas, CK7 and CK19 were positive in all cases and the expression rates of EMA, EpCAM, NCAM, CKIT, AFP, Hep Par-1 were 80.8% (21/26), 88.5% (23/26), 84.6% (22/26), 88.5% (23/26), 46.2% (12/26) and 53.8% (14/26) respectively, similar to the staining pattern of intermediate hepatocytes. In ICC areas, CK7, CK9, EMA and EpCAM were positive in all cases without the expression of NCAM and CKIT.

CONCLUSIONThe clinicopathologic findings and immunohistochemical results in this study highly suggest a hepatic progenitor cell origin of combined hepatocellular-cholangiocarcinoma (CLC type).

Bile Duct Neoplasms ; pathology ; Biomarkers ; metabolism ; Carcinoma, Hepatocellular ; pathology ; Cholangiocarcinoma ; pathology ; Hepatocytes ; cytology ; Humans ; Immunohistochemistry ; Liver Cirrhosis ; pathology ; Liver Neoplasms ; pathology ; Mucins ; metabolism ; Stem Cells ; cytology

OBJECTIVETo study the clinicopathologic and immunohistochemical features, and the prognosis of intraductal papillary mucinous neoplasms (IPMN) of the pancreas.

METHODSThe clinical findings, morphologic features, immunophenotype and prognosis were investigated in 61 cases of IPMN.

RESULTSOf these 61 cases, 33 were in the pancreatic head and 14 were in the body and tail, and 14 in the entire pancreas. The average patients' age was 61.8 years. The initial symptom was abdominal pain in 37 cases, and the tumors were detected at routine checkup in 14 cases. The imaging examination showed dilated ducts and/or cystic and solid masses. Grossly, 32 cases were multi-loculated cystic masses containing mucin and papillary areas; 13 cases were solid. Microscopically, the IPMN showed four patterns, including gastric-type (16 cases), intestinal-type (21 cases), pancreatobiliary-type (21 cases) and eosinophilic-type (3 cases). The IPMN cohort included 13, 13 and 6 IPMN with low, intermediate and high-grade dysplasia respectively, and 29 IPMN associated with invasive carcinoma. The IPMN associated carcinomas were mainly ductal adenocarcinoma (23/29, 79.3%), followed by colloid carcinoma (4/29, 13.8%) and undifferentiated carcinoma (2/29, 6.9%). Immunohistochemically, IPMN expressed MUC5AC (51/57, 89.4%), MUC2 (21/57, 36.8%), and MUC1 (13/46, 28.3%). The mean postoperative follow-up period was 32 months (range 12-112 months). Six of 61 patients were lost to follow-up. Overall 5-year survival rate was 76%. The 5-year survival rate of IPMN with low, intermediate or high-grade dysplasia was 100%, and recurrence was local in 3 patients. The 3-year survival rate of IPMN associated with invasive carcinoma was 55%. 12 of 13 patients died within 2 years after operation.

CONCLUSIONSIPMN is a common cystic neoplasm of the pancreas located in the ducts. The pathologic types and classifications are clearly defined. MUC stains are helpful for the diagnosis and papillary typing. IPMN with invasive carcinoma was associated with significantly worse survival than IPMN with dysplasia.

Adenocarcinoma, Mucinous ; diagnosis ; pathology ; Carcinoma, Pancreatic Ductal ; diagnosis ; pathology ; Humans ; Middle Aged ; Mucins ; metabolism ; Neoplasm Recurrence, Local ; Pancreas ; pathology ; Pancreatic Neoplasms ; diagnosis ; pathology ; Prognosis ; Survival Rate

Biomedical Research ; trends ; Breast Neoplasms ; metabolism ; Female ; Humans ; Mucins ; physiology ; Neoplasm Proteins ; physiology

Pigs are increasingly recognized as "natural" hosts of infection by human respiratory viruses because of their similarities to humans in terms of lung physiology, airway morphology, cell types, and distribution of cell receptors in the respiratory tract. We wished to explore the mechanisms of infection by respiratory viruses and screening of drug that could be used to treat respiratory-system diseases. Hence, we developed a model of well-differentiated porcine airway epithelial cells (PAECs) derived from pig-lung tissue and cultured them with serum-free medium under an air-liquid interface condition in vitro. We identified the PAEC model using scanning electron microscopy, electrophysiology, and immunohistology. To evaluate application of gene therapy of adeno-associated virus (AAV)6 on the PAEC model, we generated recombinant adeno-associated virus 6-green fluorescent protein (rAAV6-GFP) using the three-plasmid transfection method and infected PAECs from the apical surface with rAAV6-GFP. Results demonstrated that the PAEC model comprised a multilayer epithelial structure containing ciliated mucous secretory cells, with basal cells located directly beneath the multilayer. rAAV6-GFP could infect PAECs from the apical surface and efficiently transduce PAECs to mediate the long-term expression of the exogenous gene. Establishment of a model of well-differentiated PAECs in vitro could lay a solid foundation for the study of infection by respiratory pathogens, as well as the screening and gene therapy of agents used to treat diseases of the respiratory system.
Animals ; Cell Differentiation ; Dependovirus ; genetics ; Epithelial Cells ; cytology ; metabolism ; Green Fluorescent Proteins ; genetics ; HEK293 Cells ; Humans ; Lung ; cytology ; Membrane Potentials ; Mucins ; metabolism ; Swine ; Transduction, Genetic ; Tubulin ; metabolism

OBJECTIVETo investigate the expression of mucin 15 (MUC15) in hepatocellular carcinoma and explore its association with the prognosis of patients.

METHODSThe expression of MUC15 was detected by quantitative RT-PCR and Western blotting in liver cell line L02, liver cancer cell lines HepG2, MHCC-97H, and SMMC-7721, and in 122 HCC and corresponding adjacent non-tumor liver tissues. The association of MUC15 expression in HCC tissues with the clinical parameters and the patients' survival was analyzed.

RESULTSThe 1iver cell line L02 showed significantly higher MUC15 expression level than the liver cancer cell lines HepG2, MHCC-97H, and SMMC-7721 (P<0.05). The expression level of MUC15 was markedly lower in the HCC tissues than in the adjacent non-tumor liver tissues (P<0.05). MUC15 expression in the HCC tissues was significantly correlated with the tumor TNM stage, intrahepatic or lymphatic metastasis, portal vein thrombosis and tumor differentiation (P<0.05). Kaplan-Meier survival analysis suggested that a low MUC15 expression was associated with a poor clinical prognosis of the patients.

CONCLUSIONThe expression of MUC15 is correlated with the clinicopathological features and prognosis of HCC patients, and may potentially serve as a novel prognostic marker for HCC.

Carcinoma, Hepatocellular ; metabolism ; Cell Line, Tumor ; Hep G2 Cells ; Humans ; Kaplan-Meier Estimate ; Liver Neoplasms ; metabolism ; Lymphatic Metastasis ; Mucins ; metabolism ; Neoplasm Staging ; Prognosis

PURPOSE: Recent discoveries suggest that aberrant DNA methylation provides cancer cells with advanced metastatic properties. However, the precise regulatory mechanisms controlling metastasis genes and their role in metastatic transformation are largely unknown. To address epigenetically-regulated gene products involved in ovarian cancer metastasis, we examined the mechanisms regulating mucin 13 (MUC13) expression and its influence on aggressive behaviors of ovarian malignancies. MATERIALS AND METHODS: We injected SK-OV-3 ovarian cancer cells peritoneally into nude mice to mimic human ovarian tumor metastasis. Overexpression of MUC13 mRNA was detected in metastatic implants from the xenografts by expression microarray analysis and quantitative reverse-transcription polymerase chain reaction (qRT-PCR). The DNA methylation status within the MUC13 promoter region was determined using bisulfite sequencing PCR and quantitative methylation-specific PCR. We evaluated the effects of exogenous MUC13 on cell invasion and migration using in vitro transwell assays. RESULTS: MUC13 mRNA expression was up-regulated, and methylation of specific CpG sites within the promoter was reduced in the metastatic implants relative to those in wild-type SK-OV-3 cells. Addition of a DNA methyltransferase inhibitor to SK-OV-3 cells induced MUC13 expression, thereby implying epigenetic regulation of MUC13 by promoter methylation. MUC13 overexpression increased migration and invasiveness, compared to control cells, suggesting aberrant up-regulation of MUC13 is strongly associated with progression of aggressive behaviors in ovarian cancer. CONCLUSION: We provide novel evidence for epigenetic regulation of MUC13 in ovarian cancer. We suggest that the DNA methylation status within the MUC13 promoter region may be a potential biomarker of aggressive behavior in ovarian cancer.
Animals ; Cell Line, Tumor ; *DNA Methylation ; Epigenesis, Genetic ; Female ; *Gene Expression Regulation, Neoplastic ; Heterografts/metabolism ; Humans ; Mice ; Mice, Nude ; Mucins/*genetics/metabolism/physiology ; Neoplasm Invasiveness/genetics ; Ovarian Neoplasms/genetics/*metabolism/pathology ; RNA, Messenger/metabolism