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

OBJECTIVETo study the relationship between severity of skeletal Class III malocclusion and the patient's emotional status, as well as motivation for seeking surgical correction and satisfaction with the outcome of the surgery.

METHODSOne hundred and forty consecutive Chinese patients with skeletal Class III malocclusion who had been treated with a combined orthodontic and surgical approach were studied. Sixty-seven percent (40 males and 54 females) responded to a questionnaire. Fifty-four percent had two jaw deformities, 32% mandibular hyperplasia and 14% maxillary hypoplasia. Surgical procedures: 77% received two jaw surgeries, 15% maxillary advancement and 8% mandibular setback. This was a retrospective study based on questionnaires with numerical scale ranked answers (0: not at all; 1: a little; 2: moderately; 3: quite a bit; and 4: extremely).

RESULTSANB angle was significantly negatively correlated with feelings about the nickname related to their facial problems (embarrassment: gamma =-0.30, P < 0.01; worn out gamma =-0.32, P < 0.01; angry gamma =-0.24, P < 0.05). ANB angle also had a significant negative correlation with the reasons for having the surgery (pressure from their friends: gamma =-0.21, P < 0.05, and referred by physician: gamma =-0.24, P < 0.05). Changes in life style as a result of surgery were significantly negatively correlated with the ANB angle before treatment, positive influence on relationships with the opposite sex (gamma =-0.25, P < 0.05), positive influence in social activities (gamma =-0.22, P < 0.05).

CONCLUSIONThe psychological status before surgery and the outcome following orthognathic surgery in patients with skeletal Class III malocclusion were closely related to severity of the malocclusion.

Adolescent ; Adult ; Female ; Humans ; Jaw Abnormalities ; surgery ; Male ; Malocclusion, Angle Class II ; pathology ; psychology ; surgery ; Mandible ; abnormalities ; Maxilla ; abnormalities ; Motivation ; Retrospective Studies

Bovine theileriosis is a tick-borne disease that is hampering the development of the domestic cattle industry in northern China. This study involved a molecular survey of bovine Theileria species in 137 blood samples from cattle in the Jilin province of China. The DNA samples were screened by species-specific 18S rRNA PCR. Results revealed that 19.7% (27/137), 17.5% (24/137) and 10.9% (15/137) were found to be infected with Theileria sinensis, Theileria orientalis, respectively. Mixed infection was found in 8.8% (12/137). The overall detection rates of Baishan, Yanji, Jilin and Liaoyuan districts was 60.0%, 17.5%, 5.3% and 0%, respectively. There is little information on the detection and distribution of bovine Theileria species in northern China. Therefore, this study provides important data for understanding the epidemiology of Theileria species and designing appropriate approaches for the diagnosis and control of bovine theileriosis in northern China.

OBJECTIVE To separate and identif y the chemical constituen ts in 70％ ethanol extract of Sabia parviflora ，and to preliminarily evaluate their in vitro antioxidant activity. METHODS The chemical constituents were separated and purified by silica gel，ODS reversed-phase silica gel ，Sephadex-LH20 column and preparative high performance liquid chromatography. The structures of the isolated compounds were identified by 1H-NMR，13C-NMR and ESI-MS. The in vitro antioxidant activities of the compounds were investigated by 2，2-diphenyl-1-picrylhydrazyl radical （DPPH·），2，2′-azino-bis（3-ethylbenzothiazoline-6- sulfonate）diammonium radical （ABST+）and hydroxyl radical （OH·）. RESULTS A total of 9 compounds were isolated from the 70％ ethanol extracts of S. parviflora . They were identified as rutin （1），diiononyl phthalate （2），dibutyl phthalate （3），vomifoliol （4），rhododendrol（5），quercetin-3-O-gentiobioside（6），narcissoside（7），kaempferol-3-O-rutinoside（8）and bonaroside （9）. The in vitro antioxidant results showed that compound 1-9 showed certain in vitro antioxidant activity ，and the half scavenging concentrations of compound 1，6，7 and 8 to DPPH ·，ABST+，OH·were lower than 70 μg/mL. CONCLUSIONS Vomifoliol， rhododendrol and bonaroside are isolated from S. parviflora for the first time ，and rutin ，quercetin-3-O-gentiobioside，narcissoside and kaempferol- 3-O-rutinoside show good in vitro antioxidant activity.

OBJECTIV E To develop the infor mation-based pharmaceutical care pathway of anticoagulant therapy in patients with atrial fibrillation and improve the efficacy and safety of treatment for them. METHODS The“anticoagulant pharmaceutical care”module was developed on the basis of medical intelligent and decision system. Patients with atrial fibrillation were taken pharmaceutical care in the whole anticoagulant treatment by evaluating the thromboembolism and bleeding risks ，pre-reviewing antithrombotic prescriptions ，monitoring efficacy and drug interactions ，and warning adverse reactions. RESULTS A total of 1 228 patients receiving anticoagulant therapy were enrolled. It was found after analysis of their doctor ’s orders that 9.27％ of the patients adjusted the improper antithrombotic therapies ，3.99％ modulated treatments according to the effects of potential drug interactions or the risk of adverse reactions ，and 70.29％ of the wrong prescriptions were intervened successfully. After the information-based pharmaceutical care ，the anticoagulation treatment rate increased from 88.73％ to 97.40％，the rate of patients ’achievements to warfarin’s international normalized ratio in hospital increased from 38.64％ to 66.67％，and the incidence of serious bleeding events decreased from 2.94％ to 0.37％ （P＜0.05）. CONCLUSIONS The information-based pharmaceutical care path of anticoagulant therapy achieved comprehensive ，efficient and accurate management of patients with atrial fibrillation ，and improved the rationality ，effectiveness and safety of anticoagulant therapy.

AIMTo design and synthesize new chiral 8-(substituted) amino-analogues of 3-[(tetrahydro-2-furanyl)methyl] benzomorphans, to expand knowledge of the structure-activity relationship (SAR) for 8-aminobenzomorphan.

METHODSTarget compounds were synthesized from the 8-triflate of the optically active 3-[(tetrahydro-2-furanyl)methyl]-2,6-methano-benzomorphans using Pd-catalyzed aminations. Opioid receptor binding experiments were performed to evaluate their biological activities.

RESULTSBoth 8-amino and 8-phenylamino analogues showed lower binding affinity for mu, delta and kappa receptors than corresponding 8-hydroxy-3-[(tetrahydro-2-furanyl)methyl]-2,6-methano-benzomorphan in vitro.

CONCLUSIONThe relative poor binding affinity of the target compounds did not warrant conducting the in vivo studies to determine if they have the profile(kappa agonist/mu antagonist) that will be potentially useful in the treatment of drug addiction. Further study is in progress.

Animals ; Benzomorphans ; chemical synthesis ; chemistry ; pharmacology ; Brain ; metabolism ; Furans ; chemical synthesis ; chemistry ; pharmacology ; Guinea Pigs ; Molecular Structure ; Narcotic Antagonists ; chemical synthesis ; chemistry ; pharmacology ; Radioligand Assay ; Receptors, Opioid ; metabolism ; Receptors, Opioid, delta ; metabolism ; Receptors, Opioid, kappa ; metabolism ; Receptors, Opioid, mu ; metabolism ; Structure-Activity Relationship

OBJECTIVETo investigate the effect of interleukin-13 (IL-13) on mucus secretion in vivo and the possible mechanism.

METHODSThe SD rats were randomly divided into control group, IL-13 group and IL-13 plus SP600125 group. The phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2), c-Jun N-terminal kinase 1/2 (JNK1/2) and the level of MUC5AC in the lung tissues were examined using Western blotting. RT-PCR was performed to examine the mRNA level of STAT4 and STAT6, and electrophoretic mobility shift assays (EMSA) was used to detect the DNA-binding activities of Forkhead box a2 (FOXA2) and activator protein-1 (AP-1).

RESULTSIL-13 caused a significant increase in MUC5AC and p-JNK1/2 expression, but did not affect the phosphorylation of ERK1/2. The expression of MUC5AC was attenuated after treatment with SP600125. A significant increase in STAT6 was observed in IL-13 group compared with that in the control group, whereas the expression of STAT4 mRNA was not significantly affected. The DNA-binding activity of FOXA2 was down-regulated after IL-13 exposure, which did not affect the DNA-binding activity of AP-1.

CONCLUSIONIL-13 down-regulates mucus secretion via STAT6-FOXA2 pathway in vitro.

Animals ; Bronchi ; secretion ; Female ; Hepatocyte Nuclear Factor 3-beta ; genetics ; metabolism ; Interleukin-13 ; pharmacology ; Male ; Mucin 5AC ; metabolism ; Mucus ; secretion ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; STAT6 Transcription Factor ; genetics ; metabolism ; Signal Transduction ; drug effects

Scutellarin is a flavonoid extracted from a traditional Chinese herb, Erigeron breviscapus. The present study investigated the effect of scutellarin on MUC5AC mucin production and the possible mechanism. Human bronchial epithelial 16 (HBE16) cells were pretreated with scutellarin for 60 min, and then exposed to human neutrophil elastase (HNE) or interleukin (IL)-13 for 12 hr. RT-PCR and ELISA were performed to measure the amount of MUC5AC mucin production. The results showed that scutellarin inhibited MUC5AC expression both in mRNA and protein level induced by HNE in a concentration-dependent manner. However, scutellarin failed to inhibit MUC5AC mucin production induced by IL-13. To investigate the intracellular mechanisms associated with the effect of scutellarin on MUC5AC mucin production, western blotting was carried out to examine the phosphorylation of protein kinase C (PKC), signal transducer and activator of transcription 6 (STAT6) and extracellular signal-regulated kinase 1/2 (ERK1/2). The phosphorylation of PKC and ERK1/2 was attenuated after treatment with scutellarin, whereas STAT6 was not significantly affected. Therefore, it is suggested that scutellarin down-regulates MUC5AC mucin production on HBE16 cells via ERK-dependent and PKC-dependent pathways.
Apigenin/chemistry/*pharmacology ; Cells, Cultured ; Dose-Response Relationship, Drug ; Down-Regulation ; Epithelial Cells/*drug effects/metabolism ; Erigeron/chemistry ; Glucuronic Acids/chemistry/*pharmacology ; Humans ; Interleukin-13/*pharmacology ; Leukocyte Elastase/*pharmacology ; Mitogen-Activated Protein Kinase 1/metabolism ; Mitogen-Activated Protein Kinase 3/metabolism ; Mucin 5AC/genetics/*metabolism ; Phosphorylation ; Protein Kinase C/metabolism ; Respiratory Mucosa/drug effects/*metabolism ; STAT6 Transcription Factor/metabolism ; Signal Transduction

To determine the chemical composition of Galla chinensis extract (GCE) by several analysis techniques and to compare the efficacy of GCE and its main component(s) in inhibition of enamel demineralization, for the development of future anticaries agents, main organic composition of GCE was qualitatively determined by liquid chromatography-time of flight-mass spectrometry (LC-TOF-MS) and quantified by high-performance liquid chromatography-diode array detector (HPLC-DAD). Inorganic ions were tested by inductively coupled plasma-atomic emission spectroscopy and F was especially measured by ion chromatography. Then, bovine enamel blocks were randomly divided into four treatment groups and were subjected to a pH-cycling regime for 12 times. Each cycle included 5-min applications with one of four treatments: 4 g⋅L(-1) GCE solution, 4 g⋅L(-1) gallic acid (GA) solution, 1 g⋅L(-1) NaF solution (positive control), deionized water (DDW, negative control), and then 60-min application in pH 5.0 acidic buffer and 5-min application in neutral buffer. Acidic buffers were retained for calcium analysis. The main organic composition of GCE were GA and its isomer, and, to a lesser extent, small molecule gallotannins. The content of GA in GCE was 71.3%±0.2% (w/w). Inorganic ions were present in various amounts, of which Ca was (136±2.82) µg⋅g(-1), and Zn was (6.8±0.1) µg⋅g(-1). No F was detected in GCE. In pH cycling, GA showed an effect similar to GCE in inhibiting enamel demineralization (P>0.05). GA was found to be the main effective, demineralization inhibiting component of GCE and could be a promising agent for the development of anticaries agents.
Animals ; Calcium ; analysis ; Cariostatic Agents ; therapeutic use ; Cattle ; Chromatography, High Pressure Liquid ; Chromatography, Liquid ; Dental Enamel ; Drugs, Chinese Herbal ; chemistry ; therapeutic use ; Gallic Acid ; analysis ; therapeutic use ; Hydrolyzable Tannins ; analysis ; Mass Spectrometry ; Polyphenols ; analysis ; Random Allocation ; Tooth Demineralization ; prevention & control

Mammalian target of rapamycin (mTOR) is aberrantly activated in many cancer types, and two rapamycin derivatives are currently approved by the Food and Drug Administration (FDA) of the United States for treating renal cell carcinoma. Mechanistically, mTOR is hyperactivated in human cancers either due to the genetic activation of its upstream activating signaling pathways or the genetic inactivation of its negative regulators. The tumor suppressor liver kinase B1 (LKB1), also known as serine/threonine kinase 11 (STK11), is involved in cell polarity, cell detachment and adhesion, tumor metastasis, and energetic stress response. A key role of LKB1 is to negatively regulate the activity of mTOR complex 1 (mTORC1). This review summarizes the molecular basis of this negative interaction and recent research progress in this area.
AMP-Activated Protein Kinases ; metabolism ; Adenocarcinoma ; drug therapy ; metabolism ; Animals ; Antibiotics, Antineoplastic ; therapeutic use ; Disease Models, Animal ; Endometrial Neoplasms ; drug therapy ; metabolism ; Female ; Humans ; Mechanistic Target of Rapamycin Complex 1 ; Multiprotein Complexes ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Protein-Serine-Threonine Kinases ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Signal Transduction ; Sirolimus ; therapeutic use ; TOR Serine-Threonine Kinases ; metabolism ; Tumor Suppressor Proteins ; metabolism