PURPOSE: This study aims to investigate the additive effect of the Hedera helix (HH) and Rhizoma coptidis (RC) extracts mixture on antitussive and expectorant activities in animals. MATERIALS AND METHODS: The expectorant assay was performed with phenol red secretion in mice trachea. Mice or guinea pigs were randomly divided into groups of 8 each, including negative and positive control groups. After gastric administration of the test extracts in mice, 2.5% phenol red solution (0.2 mL) was intraperitoneally injected. Trachea was dissected and optical density of tracheal secretion was measured. After gastric administration of the test extracts in guinea pigs, the antitussive activities were assessed using a citric acid-induced cough measurement. RESULTS: The extracts of HH and RC significantly increased tracheal secretion and inhibited cough. The mixture of HH and RC extracts in a 1:1 concentration at a dose of 200 mg/kg showed a more potent effect on phenol red secretion (25.25+/-3.14) and cough inhibition (61.25+/-5.36) than the individual use of each extracts [phenol red secretion; HH 13.39+/-4.22 (p=0.000), RC 20.78+/-2.50 (p=0.010), cough inhibition; HH 9.89+/-4.14 (p=0.010), RC 30.25+/-7.69 (p=0.000)]. A 3:1 ratio mixture of HH to RC demonstrated an optimal expectorant effect (p<0.001), and this mixture showed expectorant and antitussive effects in a dose-dependent manner. CONCLUSION: This study provides evidence for antitussive and expectorant effect of a 3:1 mixture of HH and RC, which may be a useful therapeutic option for respiratory diseases.
Animals ; Antitussive Agents/*administration & dosage/pharmacology/therapeutic use ; *Behavior, Addictive ; Cough/*drug therapy ; Drugs, Chinese Herbal/*administration & dosage/pharmacology/therapeutic use ; Ethanol ; Expectorants/*administration & dosage/pharmacology/therapeutic use ; Guinea Pigs ; Hedera/*chemistry/metabolism ; Male ; Mice ; Phytotherapy ; Plant Extracts/*pharmacology ; Plant Roots/chemistry ; Trachea/drug effects/metabolism

Pulmonary tuberculosis (PTB) is a common infectious disease worldwide. However, mediastinal tuberculous lymphadenitis complicated by oesophageal involvement and oesophago-respiratory fistula is now uncommon due to improved anti-tuberculous regimes and better general awareness. The overall incidence of acquired oesophago-respiratory fistula due to infection is low, and therefore, the lesion is not often a frontrunner in differential diagnosis. Still, tuberculous oesophago-respiratory fistulae can potentially occur in patients with retroviral disease, as they tend to have atypical and more virulent manifestations. In this study, we report the case of multiple oesophago-respiratory fistulae in a patient with PTB and retroviral disease, and highlight the computed tomography features of these lesions as an atypical presentation of PTB in retroviral disease. Clinicians should suspect oesophago-respiratory fistulae if patients present with Ono’s sign, and remain particularly vigilant for patients with underlying PTB and retroviral disease, as early diagnosis and treatment could help to reduce mortality.
Adult ; Diagnosis, Differential ; Esophagus ; physiopathology ; Fistula ; diagnosis ; Humans ; Lung ; pathology ; Male ; Radiography, Thoracic ; Retroviridae ; metabolism ; Tomography, X-Ray Computed ; Trachea ; physiopathology ; Treatment Outcome ; Tuberculosis, Lymph Node ; Tuberculosis, Pulmonary ; complications ; diagnosis

PURPOSE: This study was performed to evaluate the long-term effects and safety of intratracheal (IT) transplantation of human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs) in neonatal hyperoxic lung injury at postnatal day (P)70 in a rat model. MATERIALS AND METHODS: Newborn Sprague Dawley rat pups were subjected to 14 days of hyperoxia (90% oxygen) within 10 hours after birth and allowed to recover at room air until sacrificed at P70. In the transplantation groups, hUCB-MSCs (5x10(5)) were administered intratracheally at P5. At P70, various organs including the heart, lung, liver, and spleen were histologically examined, and the harvested lungs were assessed for morphometric analyses of alveolarization. ED-1, von Willebrand factor, and human-specific nuclear mitotic apparatus protein (NuMA) staining in the lungs and the hematologic profile of blood were evaluated. RESULTS: Impaired alveolar and vascular growth, which evidenced by an increased mean linear intercept and decreased amount of von Willebrand factor, respectively, and the hyperoxia-induced inflammatory responses, as evidenced by inflammatory foci and ED-1 positive alveolar macrophages, were attenuated in the P70 rat lungs by IT transplantation of hUCB-MSCs. Although rare, donor cells with human specific NuMA staining were persistently present in the P70 rat lungs. There were no gross or microscopic abnormal findings in the heart, liver, or spleen, related to the MSCs transplantation. CONCLUSION: The protective and beneficial effects of IT transplantation of hUCB-MSCs in neonatal hyperoxic lung injuries were sustained for a prolonged recovery period without any long-term adverse effects up to P70.
Animals ; *Cord Blood Stem Cell Transplantation ; Ectodysplasins/metabolism ; Humans ; Hyperoxia/*pathology ; Lung/metabolism/pathology ; Lung Injury/pathology/*surgery ; *Mesenchymal Stem Cell Transplantation ; Models, Animal ; Nuclear Matrix-Associated Proteins/metabolism ; Rats ; Trachea/*transplantation ; von Willebrand Factor/metabolism

Aim of the present study is to investigate activation effect of nobiletin on cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel activity. CFTR-mediated iodide influx assay and patch-clamp tests were done on FRT cells stably co-transfected with human CFTR and EYFP/H148Q. Nobiletin potently activated CFTR chloride channel activity in a dose- and time-dependent manner. The CFTR blocker CFTR(inh)-172 could completely reverse the effect. Preliminary mechanism study indicated that nobiletin activated CFTR chloride channel through a direct binding way. In addition, ex vivo tests done on mice trachea showed that nobiletin time-dependently stimulated submucosal gland fluid secretion. Nobiletin may be a therapeutic lead compound in treating CFTR-related diseases including disseminated bronchiectasis.
Animals ; Benzoates ; pharmacology ; Cystic Fibrosis Transmembrane Conductance Regulator ; antagonists & inhibitors ; drug effects ; metabolism ; Dose-Response Relationship, Drug ; Epithelial Cells ; metabolism ; Exocrine Glands ; secretion ; Flavones ; administration & dosage ; pharmacology ; Humans ; Mice ; Patch-Clamp Techniques ; Rats ; Rats, Inbred F344 ; Thiazolidines ; pharmacology ; Thyroid Gland ; cytology ; Time Factors ; Trachea ; secretion

OBJECTIVETo study the analgesic, expectorant and antitussive effects of the compatible use of Aconiti Radix Cocta and Fritillaria cirrhosa or F. thunbergii with different matching ratio or dose in mice.

METHODThe two-factor, seven-level uniform design method was adopted to observe the analgesic, expectorant and antitussive effects of the oral administration with the two combined decoctions in rats, with frequency of body torsions induced by acetum, secretion of phenol red in tracheas and frequency of coughs as indexes. Significant matching proportions and doses were collected for verification.

RESULTThe effect on the frequency of body torsions: The combined decoctions could effectively reduce the frequency of body torsions. According to a regression analysis, Aconiti Radix Cocta and F. cirrhosa had the synergistic effect, which was maximized with a ratio of 1: 1. The 1: 1 combined decoction played the least role in reducing the frequency of body torsions with a total dose of more than 5 g x kg(-1). The effect on the secretion of phenol red in tracheas. The combined decoctions could effectively increase the secretion of phenol red in tracheas. According to a regression analysis, Aconiti Radix Cocta and F. thunbergii had the antagonism, which was maximized at the ratio of 1: 1, and minimized with a total dose of less than 10 g x kg(-1) and a ratio of 5: 1 between F. thunbergii and Aconiti Radix Cocta. The effect on the frequency of coughs. The combined decoctions could effectively reduce the frequency of coughs. According to a regression analysis, Aconiti Radix Cocta and F. cirrhosa had the antagonism, which was maximized at the ratio of more than 1: 5 and less than 10: 1. There was no interaction between Aconiti Radix Cocta and F. thunbergii. F. thunbergii could reduce the frequency of coughs, whereas Aconiti Radix Cocta showed no effect.

CONCLUSIONThe compatible application of Aconiti Radix Cocta and F. cirrhosa could enhance the analgesic effect of Aconiti Radix Cocta and reduce the expectorant and antitussive effects of F. cirrhosa, which vary according to different matching ratio and dose. The compatible application of Aconiti Radix Cocta and F. thunbergii shows no effect on the antitussive effect of F. thunbergii. This study provides experimental basis for in-depth studies on the combined effect of Aconiti Radix Cocta and Fritillaria--two of eighteen incompatible pairs.

Aconitum ; chemistry ; Analgesics ; pharmacology ; Animals ; Antitussive Agents ; pharmacology ; Behavior, Animal ; drug effects ; Drug Synergism ; Drugs, Chinese Herbal ; pharmacology ; Expectorants ; pharmacology ; Fritillaria ; chemistry ; Male ; Mice ; Phenolsulfonphthalein ; metabolism ; Trachea ; drug effects ; metabolism

OBJECTIVE: To construct phosphorylation sites domain (PSD) mutant of myristoylated alaninerich C kinase substrate (MARCKS) and explore the role of transient receptor potential melastatin 8 cation channels (TRPM8) and MARCKS in cold-induced synthesis and exocytosis of mucin (MUC) 5AC. METHODS: Human placental cDNA was used as a template to amplify the full coding region of MARCKS cDNA by PCR. Ser159, Ser 163, Ser 167, Ser 170 in the PSD were mutated to aspartic acids by an overlap PCR method. The resultant PSD mutant cDNA and the wild-type MARCKS cDNA were each subcloned into a mammalian expression vector pcDNA3.0. Recombinant constructs were confirmed by restriction enzyme digestion analysis and DNA sequencing. In intervention experiments, cells were pretreated with the TRPM8 channel antagonist BCTC and transfected with MARCKS-PSD mutant cDNA, and thereafter cold stimulation was applied. The levels of MUC5AC were measured by immunofluorescence and ELISA to clarify the roles of TRPM8 and PSD mutant on the synthesis and secretion of MUC5AC induced by cold, respectively. RESULTS: Restriction enzyme digestion analysis and DNA sequencing revealed that the pcDNA3.0- MARCKS and pcDNA3.0-MARCKS-PSD mutants were successfully constructed. The levels of intracellular and secreted MUC5AC of cold treated group were significantly higher than those of control group (P<0.05). BCTC attenuated the cold-induced synthesis and secretion of MUC5AC when compared with cold treated group (P<0.05). Transfection of 16HBE cells with the MARCKS-PSD mutant cDNA resulted in significant inhibition of mucin secretion in response to cold, and significantly higher level of intracellular MUC5AC than that of control group (P<0.01), whereas transfection with the vector DNA or the wild-type MARCKS cDNA had no effect on the mucin synthesis and secretion in response to cold (P>0.05). CONCLUSION TRPM8 and phosphorylation of MARCKS-PSD mediates the cold-induced exocytosis of MUC5AC by airway epithelial cells.
Base Sequence ; Cell Line ; Cold Temperature ; Epithelial Cells ; cytology ; metabolism ; Exocytosis ; physiology ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; metabolism ; Membrane Proteins ; genetics ; metabolism ; Molecular Sequence Data ; Mucin 5AC ; metabolism ; Mutation ; Myristoylated Alanine-Rich C Kinase Substrate ; Phosphorylation ; TRPM Cation Channels ; metabolism ; Trachea ; cytology ; metabolism

Recently, the world's first transgenic dogs were produced by somatic cell nuclear transfer. However, cellular senescence is a major limiting factor for producing more advanced transgenic dogs. To overcome this obstacle, we rejuvenated transgenic cells using a re-cloning technique. Fibroblasts from post-mortem red fluorescent protein (RFP) dog were reconstructed with in vivo matured oocytes and transferred into 10 surrogate dogs. One puppy was produced and confirmed as a re-cloned dog. Although the puppy was lost during birth, we successfully established a rejuvenated fibroblast cell line from this animal. The cell line was found to stably express RFP and is ready for additional genetic modification.
Animals ; Animals, Genetically Modified ; Cloning, Organism/methods/*veterinary ; Dogs/*genetics ; Female ; Gastrointestinal Tract/metabolism ; Gene Expression Regulation ; Kidney/metabolism ; Liver/metabolism ; Luminescent Proteins/*genetics/metabolism ; Lung/metabolism ; Male ; Myocardium/metabolism ; Nuclear Transfer Techniques/veterinary ; Spleen/metabolism ; Trachea/metabolism

BACKGROUNDAn important physiological feature of asthma is the phenotypic change of airway smooth muscle cells (ASMCs), but the precise mechanisms behind the ASMCs' change remains unknown. Our study assessed whether p21Ras can directly modulate the phenotype of ASMCs.

METHODSRat ASMCs were treated with FTP III, a highly specific p21Ras inhibitor. ASMCs were identified via immunocytochemistry. The ultrastructure of cells was observed by electron microscopy, and the expression of α-actin was evaluated by Western blotting analysis. The levels of IL-6 and RANTES were measured by enzyme linked immunosorbent assay (ELISA).

RESULTSIt was observed that ASMCs in asthma exhibited a proliferative/secretory phenotype and were larger, denser and had many pseudopods, as well as increased signs of secretory organelles. Additionally, the level of α-actin, a marker of ASMCs, was reduced in asthmatic ASMCs and the secretion of IL-6 and RANTES was increased. When FTP III was added to asthmatic ASMCs it induced a contractile phenotype, with increased α-actin levels and reduced secretion of IL-6 and RANTES.

CONCLUSIONSIt appears that p21Ras induces asthmatic ASMCs to a proliferative/secretory phenotype, but its inhibitor FTP III, can significantly reverse this phenotype. The role of p21Ras in the ASMCs may be a new target for asthma treatment.

Actins ; metabolism ; Animals ; Asthma ; metabolism ; Blotting, Western ; Cells, Cultured ; Chemokine CCL5 ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Interleukin-6 ; metabolism ; Lung ; cytology ; Male ; Microscopy, Electron, Transmission ; Myocytes, Smooth Muscle ; drug effects ; metabolism ; ultrastructure ; Organophosphonates ; pharmacology ; Proto-Oncogene Proteins p21(ras) ; antagonists & inhibitors ; metabolism ; Rats ; Rats, Sprague-Dawley ; Trachea ; cytology

OBJECTIVEIn this study, we pretreated the mice ASMCs by dexamethasone (Dex) within 10 min, to test the peak of [Ca2+]i and phospho-PLCbeta (ser1105) in the cells by treated with Ach.

METHODSThe peak of [Ca2+]i was measured by Fura-2/AM methods and the phospho-PLCbeta-ser1105 was by Western blot, and compared with dexamethasone pretreated groups. Glucocorticoid receptor antagonist RU486 and the protein synthesis inhibitor cycloheximide groups were settled in our study.

RESULTSGlucocorticoids (GCs) significantly decreased the resting values and peak of [Ca2+]i elevation and elevated the intracellular levels of phospho-PLCbeta (ser1105) in 10 min. Neither the RU486 nor cycloheximide could alter the inhibitory effects of glucocorticoids stated above.

CONCLUSIONOur results demonstrate that glucocorticoids exert rapid inhibitory effects. The series of signal changes in this process that restrain the peak of [Ca2+]i may be responsible for the rapid nongenomic inhibitory effects of GCs by reducing the activity of PLC.

Animals ; Calcium ; metabolism ; Cells, Cultured ; Dexamethasone ; pharmacology ; Glucocorticoids ; pharmacology ; Guinea Pigs ; Male ; Mifepristone ; pharmacology ; Muscle, Smooth ; drug effects ; metabolism ; Phospholipase C beta ; metabolism ; Rats ; Rats, Sprague-Dawley ; Trachea ; cytology

OBJECTIVETo observe the effect of repeated esophageal acid infusion on specific airway resistance (sRaw) and airway reactivity in the guinea pigs and explore the mechanism.

METHODSsRaw and airway reactivity were measured by double-chamber plethysmography in normal control group (group N), saline control group (group NS), and repeated acid irrigation group (group H). The initial measurement was used as the baseline sRaw and airway reactivity (1d1), and 2 h after the initial measurement, sRaw and airway reactivity were measured again (1d2). Similarly, such measurements were repeated on the 15th day for all the guinea pigs (15d1, 15d2) with a 2-h interval. The content of Substance P (SP) and vasoactive intestinal peptide (VIP) in lung tissue, trachea, BALF and ganglion were detected by ELISA.

RESULTSThe percent change of sRaw, (15d2-1d1)/1d1 in group H was significantly higher than that in group N. The differences in the airway reactivity of the group N, group NS, and group H were not statistically significant. The SP content in the lung, trachea, ganglion and bronchoalveolar lavage fluid (BALF) in group H was significantly higher than those in group N. The SP content in ganglion showed a significant positive correlation to that in the trachea. No significant differences were found in the VIP content in the lung, trachea, ganglion or BALF between the groups.

CONCLUSIONRepeated esophageal acid infusion increases the airway resistance, but not the airway reactivity in normal guinea pigs. SP may be involved in development of high sRaw through the esophageal-tracheobronchial reflex.

Airway Resistance ; Animals ; Bronchoalveolar Lavage Fluid ; chemistry ; Esophagus ; Gastroesophageal Reflux ; metabolism ; physiopathology ; Guinea Pigs ; Lung ; metabolism ; Male ; Respiratory System ; Substance P ; metabolism ; Trachea ; metabolism ; Vasoactive Intestinal Peptide ; metabolism