ABSTRACT
Objective:To investigate the effect of PKM2 on proliferation and apoptosis of nasopharyngeal carcinoma cells.Methods:Nasopharyngeal carcinoma cell CNE-1 was transfected with PKM2 small interfering RNA (PKM2 siRNA1 and PKM2 siRNA2) and negative controls (siRNA control),the levels of PKM2 in the cells were detected by fluorescent quantitative PCR and Western blot,screening interference PKM2 siRNA2 continued to study.Cell proliferation was detected by MTT,cell cloning test showed the ability of cell cloning,apoptosis was detected by flow cytometry,ROS level was detected by DCFH-DA,the levels of p38MAPK,p-p38MAPK,C-myc,β-catenin,Cleaved Caspase-3 protein were detected by Western blot.Results:After transfection of PKM2 siRNA1 and PKM2 siRNA2,the levels of PKM2 mRNA and protein were significantly decreased compared with those without transfection,and after transfection of PKM2 siRNA2,the level of PKM2 in cells decreased more,the levels of PKM2 in transfected siRNA control cells were not significantly different from those without transfection.The rate of apoptosis after down-regulation of PKM2 expression increased from (9.36 ± 1.04)% to (48.42 ± 5.28)%,and the rate of cell clone formation decreased from (75.48 ± 8.25)% to (46.15 ± 3.47)%,OD values from (0.86±0.11) down to (0.52±0.04),elevated levels of ROS in cells,the levels of p-p38MAPK,Cleaved Caspase-3 proteins in cells were also significantly increased, the levels of C-myc and β-catenin in cells were obviously decreased.Conclusion:Downregulation of PKM2 expression inhibits nasopharyngeal carcinoma cell growth,promoting apoptosis of naso-pharyngeal carcinoma cells,the mechanism of action may be related to p38MAPK and Wnt/β-catenin signaling pathway.
ABSTRACT
Purpose To investigate the clinicopathologic characteristtics, immunophenotype and differential diagnosis of uterine adenosarcoma with sarcomatous overgrowth. Metheds The clinicopathological data of 4 cases of uterine adenosarcoma with sarcomatous overgrowth were collected, the histopathologic and immunohistochemical features were investigated, and the rele-vant literatures were also reviewed. Results All of tumors were arised from the endometrium with complains of postmenopausal vaginal bleeding or prolonged menstrual period. There is a poly-poid nodular in the uterine cavity with a pedicle or no pedicel, or rough endometrium. On the cut surface, the tumor was fish-like without distinct from the surrounding tissue. Light microsco-py show the tumors were composed of benign glands and malignant mesenchymal components, the sarcomatouscomponents ac-counted for over 25％. In 4 cases, 2 cases had heterologous com-ponent of rhabdomyosarcoma. The component of sarcomatous were positive for vimentin and CD10. The heterologous component of rhabdomyosarcoma were positive for desmin, MyoDl, and Myogenin.3 cases were died at in 5, 10, and 19 months after operation, 1 patient was disease free survival for 3 months. Conclusion Uterine adenosarcoma with sarcomatous overgrowth has a bad prognosis.
ABSTRACT
Airway remodeling is an important pathological feature of asthma and the basis of severe asthma. Proliferation of airway smooth muscle cells (ASMCs) is a major contributor to airway remodeling. As an important Ca(2+) channel, transient receptor potential vanilloid 1 (TRPV1) plays the key role in the cell pathological and physiological processes. This study investigated the expression and activity of TRPV1 channel, and further clarified the effect of TRPV1 channel on the ASMCs proliferation and apoptosis in order to provide the scientific basis to treat asthmatic airway remodeling in clinical practice. Immunofluorescence staining and reverse transcription polymerase chain reaction (RT-PCR) were used to detect the expression of TRPV1 in rat ASMCs. Intracellular Ca(2+) was detected using the single cell confocal fluorescence microscopy measurement loaded with Fluo-4/AM. The cell cycles were observed by flow cytometry. MTT assay and Hoechst 33258 staining were used to detect the proliferation and apoptosis of ASMCs in rats respectively. The data showed that: (1) TRPV1 channel was present in rat ASMCs. (2) TRPV1 channel agonist, capsaicin, increased the Ca(2+) influx in a concentration-dependent manner (EC50=284.3±58 nmol/L). TRPV1 channel antagonist, capsazepine, inhibited Ca(2+) influx in rat ASMCs. (3) Capsaicin significantly increased the percentage of S+G2M ASMCs and the absorbance of MTT assay. Capsazepine had the opposite effect. (4) Capsaicin significantly inhibited the apoptosis, whereas capsazepine had the opposite effect. These results suggest that TRPV1 is present and mediates Ca(2+) influx in rat ASMCs. TRPV1 activity stimulates proliferation of ASMCs in rats.
ABSTRACT
The proliferation of cardiac fibroblasts (CFs) is a key pathological process in the cardiac remodeling. To establish an objective, quantitative method for the analysis of cell proliferation and cell cycle, we applied the high-content screening (HCS) and flow cytometry (FCM) techniques. CFs, isolated by enzyme digestion from newborn C57BL/6J mice, were serum starved for 12 h and then given 10% fetal bovine serum (FBS) for 24 h. Followed by BrdU and DAPI (or 7-AAD) staining, CFs proliferation and cell cycle were analyzed by HCS and FCM, respectively. Discoidin domain receptor 2 (DDR2) staining indicated that the purity of isolated CFs was over 95%. (1) HCS analysis showed that the ratio of BrdU-positive cells was significantly increased in 10% FBS treated group compared with that in serum-free control group [(12.96 ± 0.67)% vs (2.77 ± 0.33)%; P < 0.05]. Cell cycle analysis showed that CFs in G0/G1 phase were diploid, and CFs in S phase were companied with proliferation, DNA replication and enlarged nuclei; CFs in G2 phase were tetraploid, and CFs in M phase produced two identical cells (2N). (2) FCM analysis showed that the ratio of BrdU-positive cells was increased in 10% FBS treated group compared with that in the control group [(11.10 ± 0.42)% vs (2.22 ± 0.31)%; P < 0.05]; DNA content histogram of cell cycle analysis indicated that the platform of S phase elevated in 10% FBS group compared with control group. (3) There were no differences between the two methods in the results of proliferation and cell cycle analysis. In conclusion, HCS and FCM methods are reliable, stable and consistent in assessment of the proliferation and cell cycle in CFs.
Subject(s)
Animals , Mice , Cell Cycle , Cell Proliferation , Fibroblasts , Cell Biology , Flow Cytometry , Mice, Inbred C57BL , Mitosis , Myocardium , Cell BiologyABSTRACT
Airway remodeling is an important pathological feature of asthma and the basis of severe asthma. Proliferation of airway smooth muscle cells (ASMCs) is a major contributor to airway remodeling. As an important Ca(2+) channel, transient receptor potential vanilloid 1 (TRPV1) plays the key role in the cell pathological and physiological processes. This study investigated the expression and activity of TRPV1 channel, and further clarified the effect of TRPV1 channel on the ASMCs proliferation and apoptosis in order to provide the scientific basis to treat asthmatic airway remodeling in clinical practice. Immunofluorescence staining and reverse transcription polymerase chain reaction (RT-PCR) were used to detect the expression of TRPV1 in rat ASMCs. Intracellular Ca(2+) was detected using the single cell confocal fluorescence microscopy measurement loaded with Fluo-4/AM. The cell cycles were observed by flow cytometry. MTT assay and Hoechst 33258 staining were used to detect the proliferation and apoptosis of ASMCs in rats respectively. The data showed that: (1) TRPV1 channel was present in rat ASMCs. (2) TRPV1 channel agonist, capsaicin, increased the Ca(2+) influx in a concentration-dependent manner (EC50=284.3±58 nmol/L). TRPV1 channel antagonist, capsazepine, inhibited Ca(2+) influx in rat ASMCs. (3) Capsaicin significantly increased the percentage of S+G2M ASMCs and the absorbance of MTT assay. Capsazepine had the opposite effect. (4) Capsaicin significantly inhibited the apoptosis, whereas capsazepine had the opposite effect. These results suggest that TRPV1 is present and mediates Ca(2+) influx in rat ASMCs. TRPV1 activity stimulates proliferation of ASMCs in rats.
Subject(s)
Animals , Rats , Antipruritics , Pharmacology , Apoptosis , Physiology , Bronchi , Cell Biology , Metabolism , Calcium Signaling , Physiology , Capsaicin , Pharmacology , Cell Proliferation , Myocytes, Smooth Muscle , Cell Biology , Metabolism , Rats, Sprague-Dawley , TRPV Cation Channels , MetabolismABSTRACT
Bronchial asthma is a common chronic airway inflammatory disease. Asthma is associated with high mortality, especially in the elderly patients. Repeated exacerbations cause disease progression. Therefore, identifying the onset of acute elderly asthma as soon as possible and giving the effective treatment is crucial to improve the prognosis. This study was to investigate the significance of fractional exhaled nitric oxide (FeNO) combined with serum procalcitonin (PCT) and C-reactive protein (CRP) in the evaluation of elderly asthma. A total of 120 elderly patients with an acute attack of asthma from July, 2010 to May, 2012 were studied. On presentation, FeNO, serum PCT and CRP concentrations were measured and sputum culture was also performed. The elderly patients were re-evaluated when they had returned to their stable clinical state. The elderly patients were classified into two groups: positive bacterial culture group (A) and negative bacterial culture group (B). The results showed that: (1) In patients with an acute exacerbation of asthma, 48 (40%) patients had positive sputum bacterial culture and 72 (60%) had negative sputum bacterial culture. (2) The levels of FeNO in patients with acute exacerbation of asthma were significantly higher than in those with no acute exacerbation state (63.8±24.6 vs. 19±6.5 ppb, P0.05). (3) The levels of PCT and CRP in group A patients with an acute exacerbation of asthma were significantly higher (P0.05) when compared with the exacerbation group. There were no significant differences in the levels of PCT and CRP between the two groups in non-acute exacerbation state (P>0.05). These results suggest that the increase in FeNO indicates the acute exacerbation of asthma, and the elevation of serum PCT and CRP levels may be associated with bacterial infection.
ABSTRACT
Bronchial asthma is a common chronic airway inflammatory disease. Asthma is associated with high mortality, especially in the elderly patients. Repeated exacerbations cause disease progression. Therefore, identifying the onset of acute elderly asthma as soon as possible and giving the effective treatment is crucial to improve the prognosis. This study was to investigate the significance of fractional exhaled nitric oxide (FeNO) combined with serum procalcitonin (PCT) and C-reactive protein (CRP) in the evaluation of elderly asthma. A total of 120 elderly patients with an acute attack of asthma from July, 2010 to May, 2012 were studied. On presentation, FeNO, serum PCT and CRP concentrations were measured and sputum culture was also performed. The elderly patients were re-evaluated when they had returned to their stable clinical state. The elderly patients were classified into two groups: positive bacterial culture group (A) and negative bacterial culture group (B). The results showed that: (1) In patients with an acute exacerbation of asthma, 48 (40%) patients had positive sputum bacterial culture and 72 (60%) had negative sputum bacterial culture. (2) The levels of FeNO in patients with acute exacerbation of asthma were significantly higher than in those with no acute exacerbation state (63.8±24.6 vs. 19±6.5 ppb, P<0.05). There was no significant difference in FeNO between group A and group B (P>0.05). (3) The levels of PCT and CRP in group A patients with an acute exacerbation of asthma were significantly higher (P<0.05) than in group B (for PCT: 27.46±9.32 vs. 7.85±3.52 ng/mL; for CRP: 51.25±11.46 vs. 17.11±5.87 mg/L, respectively). When they had returned to stable clinical state, the levels of PCT and CRP in group A were decreased significantly (P<0.05), and those in group B had no significant change (P>0.05) when compared with the exacerbation group. There were no significant differences in the levels of PCT and CRP between the two groups in non-acute exacerbation state (P>0.05). These results suggest that the increase in FeNO indicates the acute exacerbation of asthma, and the elevation of serum PCT and CRP levels may be associated with bacterial infection.
Subject(s)
Aged , Female , Humans , Male , Asthma , Diagnosis , Metabolism , Biomarkers , Metabolism , Breath Tests , Methods , C-Reactive Protein , Metabolism , Calcitonin , Blood , Calcitonin Gene-Related Peptide , Exhalation , Nitric Oxide , Metabolism , Protein Precursors , Blood , Reproducibility of Results , Sensitivity and SpecificityABSTRACT
<p><b>OBJECTIVE</b>To explore the inhibitory effect of combined administration of bear bile powder (BBP) and cyclophosphamide (Cytoxan, CTX) on colorectal cancer liver metastasis by regulating tumor promotion inflammation microenvironment.</p><p><b>METHOD</b>The CRC liver metastasis mode in mice was established through in situ spleenic injection of SL4 tumor cells into spleens. The mice were randomly divided into 5 groups: the model group, the CTX (80 mg x kg(-1)) treatment group, the CTX + BBP high dose (300 mg x kg(-1)) group, the CTX + BBP middle dose (150 mg x kg(-1)) group and the CTX + BBP low dose (75 mg x kg(-1)) group. Mice were orally administered with drugs for 12 days, and sacrificed on the 13'h day for weighing their spleens and lives, HE staining, and immunofluorescence analysis. Their peripheral blood, and metastatic tumor in spleens and lives were analyzed with flow cytometry.</p><p><b>RESULT</b>Spleen and liver weights of the: CTX treatment group and other doses groups were significantly lower than that of the model group. HE staining and immunofluorescence analysis showed that lymphocyte infiltration was detected in normal tissues, and macrophages infiltration was observed around the tumor tissues. Flow cytometry analysis showed that the number of T-lymphocytes in peripheral blood of different doses groups were much higher than that of the CTX treatment group (P < 0.05), with the rise in the ratio of CD4/CD8; the total number of lymphocytes in spleen cell suspension increased in different doses groups, compared to the CTX treatment group, with notable increase in B cells (P < 0.05) and significant decrease in CD11b, F4/80 cells (P < 0.05). The combined treatment showed less monocyte macrophages in liver metastasis than that of the CTX treatment group.</p><p><b>CONCLUSION</b>The combined treatment of bear bile powder and cyclophosphamide has the effect in not only protecting liver and increase immunity, but also in anti-inflammation and antitumor by regulating tumor microenvironment and reducing the collection of mononuclear macrophages. Particularly, the combined administration of low dose of bear bile powder and CTX shows the most significant effect in reducing inflammatory cell infiltration.</p>
Subject(s)
Animals , Humans , Male , Mice , Bile , Chemistry , Colorectal Neoplasms , Drug Therapy , Mortality , Pathology , Combined Modality Therapy , Cyclophosphamide , Liver Neoplasms , Drug Therapy , Mortality , Mice, Inbred C57BL , Tumor Microenvironment , UrsidaeABSTRACT
Objective To compare insulin secretion and action with impaired fasting glucosc (IFG),impaircd glucose tolerance (IGT) and combined glucose intolerance (CGI,IFG and IGT) between Han and Uygur populations living in Xinjiang.Methods A multicenter cross-section survey (The Third Diabetes Epidemiological Survey in China) was conductcd in Xinjiang from 2007 to 2008 including 2203 subjects (Han 1118,Uygur 1085) underwent an oral glucosc test (OGTT).Homeostasis model assessment on insulin resistance (HOMA-IR) and β cell function (HOMA-β)were calculated.The ratio of incrcmcntal insulin(Δ130 ) and glucose (ΔG30)response was used to evaluate the early insulin secretion.ΔI30/ΔG30/HOMA-IR was used to evaluate the glucosc disposition index (DI).Results There were differences noticed regarding the waist circumstances (WC),body mass index (BMI),lipids,0 and 120 min insulin lcvcls in different glucose tolerance status between the Hans and Uygurs.Data related to NGT,IFG,CGI,WC from the Uygurs was significantly diffcrcnt from that of the Hans (P＜0.01),while the NGT,IFG,IGT and 120-minute plasna insulin levels of the Hans were significantly different from that of the Uygurs (P＜0.01).HOMA-IR and HOMA-β in Hans were significantly different from those of the Uygurs (P＜0.01).There were significant differences noticed on data reoated to Δ130/ΔG30,and DI among the two populations with different ethnicities.Conclusion Regarding the regulation of impaired glucose,the insulin resistance among the Hans was significantly different from that of the Uygurs,while there seemed to be a compensatory secretion of pancreatic β cells which played the role of maintaining blood glucose homeostasis.
ABSTRACT
The paper is to report the establishment of three methods for determination of methyl salicylate-2-O-beta-D-galactopyranoside (1-4)-beta-D-glucopyranoside (MSG) by HPLC, UV or potentiometric titration. The results determined by the three methods turned out to be of no significant difference (P>0.05). The method was chosen according to MSG difference test demands.
Subject(s)
Anti-Inflammatory Agents , Chemistry , Chromatography, High Pressure Liquid , Methods , Glycosides , Chemistry , Molecular Structure , Potentiometry , Methods , Reproducibility of Results , Salicylates , Chemistry , Sensitivity and Specificity , Spectrophotometry, Ultraviolet , MethodsABSTRACT
<p><b>OBJECTIVE</b>To investigate the relationship between the proliferation of sensitized human airway smooth muscle cells (HASMCs) and the expression of extracellular signal regulated kinase (ERK) and the effect of Shenmai Injection (SMI) on HASMCs.</p><p><b>METHODS</b>The HASMCs cultured in vitro were divided into three groups: (1) control group; (2) sensitized group: containing 10% asthmatic serum; (3) SMI group: further divided into three different concentration subgroups interferred with 10 microL/mL, 50 microL/mL, and 100 microL/mL SMI, respectively. The proliferation of HASMCs was detected using MTT method, the expression of proliferating cell nucleus antigen (PCNA) in HASMCs was detected using immunocytochemical staining, and the expression of phosphoration-ERK1/2 (p-ERK1/2) protein was detected using Western-blot.</p><p><b>RESULTS</b>After passive sensitization,: the optical density value (A A(490) value) of HASMCs was significantly increased from 0.366+/-0.086 to 0.839+/- 0.168 (P<0.05). In addition, the expression of PCNA was significantly increased from 28.7%+/-5.9% in the control group to 69.8%+/-7.5% in the sensitized group (P<0.05). At the same time, the expression of p-ERK1/2 in passively sensitized HASMCs was significantly increased compared with the control group (all P<0.05). After application of 10 microL/mL, 50 microL/mL, and 100 microL/mL SMI to the cultured media of passively sensitized group, the A(570) value was significantly decreased from 0.839+/-0.168 to 0.612+/-0.100, 0.412+/-0.092, and 0.339+/-0.077, respectively (P<0.05). Moreover, the expression of PCNA was significantly decreased from 69.8%+/-7.5% to 57.8%+/-6.2%, 40.7%+/-5.4%, and 26.1%+/-5.2%, respectively. At the same time, the expression of p-ERK1/2 in each SMI group was significantly decreased compared with the sensitized group (all P<0.05).</p><p><b>CONCLUSION</b>ERK signal transduction pathway may be involved in the airway remodeling in asthma. The expression of ERK can be inhibited by SMI in a dose-dependent manner, thus preventing the proliferation of HASMCs.</p>
Subject(s)
Adolescent , Adult , Female , Humans , Male , Middle Aged , Young Adult , Asthma , Pathology , Blotting, Western , Cell Proliferation , Drug Combinations , Drugs, Chinese Herbal , Pharmacology , Extracellular Signal-Regulated MAP Kinases , Metabolism , Injections , Myocytes, Smooth Muscle , Pathology , Proliferating Cell Nuclear Antigen , MetabolismABSTRACT
<p><b>BACKGROUND</b>Current prosthetic, small diameter vascular grafts showing poor long term patency rates have led to the pursuit of other biological materials. Biomaterials that successfully integrate into surrounding tissue should match not only the mechanical properties of tissues, but also topography. Polyglycolic acid (70/30) has been used as synthetic grafts to determine whether human vascular smooth muscle cells and endothelial cells attach, survive and secrete endothelin and 6-keto-prostaglandin F1alpha (6-keto-PGF1alpha).</p><p><b>METHODS</b>Endothelial cells and smooth muscle cells were isolated from adult human great saphenous vein. They were seeded on polyglycolic acid scaffold in vitro separately to grow vascular patch (Groups A and B respectively) and cocultured in vitro to grow into vascular patch (Group C). Smooth muscle cells and endothelial cells were identified by immunohistochemical analysis and growth of cells on polyglycolic acid was investigated using scanning electron microscopy. The levels of endothelin and 6-keto-PGF1alpha in the culturing solutions were examined by radioimmunology to measure endothelial function.</p><p><b>RESULTS</b>Seed smooth muscle cells adhered to polyglycolic acid scaffold and over 28 days grew in the interstices to form a uniform cell distribution throughout the scaffold. Then seed endothelial cells formed a complete endothelial layer on the smooth muscle cells. The levels of endothelin and 6-keto-prostaglandin F1 alpha in the culturing solution were (234 +/- 29) pg/ml and (428 +/- 98) pg/ml respectively in Group C and (196 +/- 30) pg/ml and (346 +/- 120) pg/ml in Group B; both significantly higher than in Groups A and D (blank control group, all P < 0.05).</p><p><b>CONCLUSIONS</b>Cells could be grown successfully on polyglycolic acid and retain functions of secretion. Our next step is to use human saphenous vein smooth muscle cells and endothelial cells to grow tubular vascular grafts in vitro.</p>
Subject(s)
Adult , Humans , 6-Ketoprostaglandin F1 alpha , Blood Vessel Prosthesis , Coculture Techniques , Endothelial Cells , Physiology , Muscle, Smooth, Vascular , Cell Biology , Myocytes, Smooth Muscle , Physiology , Polyglycolic Acid , Pharmacology , Saphenous Vein , Cell Biology , Tissue EngineeringABSTRACT
<p><b>OBJECTIVE</b>To investigate the novel hyperplasia suppressor gene (HSG) expression in vascular smooth muscle cells derived from normotensive and hypertensive patients underwent bypass surgery.</p><p><b>METHODS</b>Coronary heart disease patients underwent coronary artery bypass graft (CABG) operation in BEIJING ANZHEN hospital from 4 - 9, 2006 were enrolled in this study and divided into hypertensive group (n = 28) and normotensive group (n = 26). The preoperative venous blood samples were taken for serum biochemical and vasoactive peptides measurements. Total RNA was extracted from WBC, explanted-vessels and cultured VSMCs using TRIZOL and HSG expression was determined by Semi-Quantitative RT-PCR.</p><p><b>RESULTS</b>Body mass index (BMI) was significantly higher in hypertensive group compared to normotensive group (P < 0.01) while other biochemic parameters and vasoactive peptides were similar between the groups. BMI and GLU, BMI and SBP, BMI and DBP, GLU and TG, SBP and DBP were positively correlated (all P < 0.05). HSG expression in WBC, VSMCs and vessel tissue were significantly lower in hypertensive group than those in normotensive group (all P < 0.05). HSG expression in tissue was negatively correlated to BMI, SBP and DBP (all P < 0.05).</p><p><b>CONCLUSIONS</b>Reduced HSG expression and the negative correlation on vascular tissue HSG expression to BMI, SBP and DBP suggested a possible inhibitory role of HSG on VSMC proliferation and blood pressure.</p>
Subject(s)
Aged , Female , Humans , Male , Middle Aged , Body Mass Index , Gene Expression , Genes, Suppressor , Hyperplasia , Genetics , Hypertension , Genetics , Pathology , Muscle, Smooth, Vascular , Cell Biology , Myocytes, Smooth Muscle , MetabolismABSTRACT
To determine the possible roles of survivin in the pathogenesis of myelodysplastic syndrome (MDS) and to explore the relationship between apoptosis and angiogenesis in MDS, the expressions of survivin, Bcl-2 and VEGF were detected in the BM cells of de novo patients with MDS, patients with AML and individuals of control by immunochemical staining and their relationship was analyzed. The results showed that the expression rate and integral of all the three proteins in the low-risk group of MDS, high-risk group of MDS and de novo acute myeloid leukemia patients gradually increased, in addition to expression of Bcl-2 in low-risk group of MDS and control group. The significant differences were observed in every two groups and there were positive relations between the every two proteins. It is concluded that survivin, Bcl-2 and VEGF are all involved in the pathogenesis of MDS, and related with the progression of this disease, the deregulated apoptosis and angiogenesis may be involved in the pathogenesis of MDS through interaction among three proteins mentioned above.
Subject(s)
Adult , Female , Humans , Male , Apoptosis , Physiology , Bone Marrow Cells , Metabolism , Pathology , Inhibitor of Apoptosis Proteins , Microtubule-Associated Proteins , Genetics , Myelodysplastic Syndromes , Metabolism , Pathology , Neoplasm Proteins , Genetics , Neovascularization, Pathologic , Proto-Oncogene Proteins c-bcl-2 , Genetics , Vascular Endothelial Growth Factor A , GeneticsABSTRACT
<p><b>AIM</b>To investigate the role of delayed rectifier K+ channel (Kv), Ca2+ -activated K+ channel (K(Ca)) and ATP-sensitive K+ channel (K(ATP)) in the regulation of the resting and contracting tone of human control and passively sensitized bronchial smooth muscle (BSM).</p><p><b>METHODS</b>The regulating effects of the three K+ channels on the tone of human BSM (HBSM) were observed by measuring the isometric tone of bronchial rings in vitro.</p><p><b>RESULTS</b>(1) The contraction of passively sensitized bronchial ring was significantly increased by histamine. (2) Kv blocker 4-aminopyridine (4-AP) caused concentration dependent contraction in resting bronchial rings of two groups, and the contraction sensitivity of the sensitized group rings was significantly stronger than that of control, that is, the negative logarithm of the drug concentration causing 50% of maximal effect (pD2) of the sensitized group rings were significantly larger than that of control rings, but there was no difference in the maximal effect (Emax) of two groups; Kca blocker tetraethylammonium (TEA) and K(ATP) blocker glibenclamide (Glib) had no such effects as those of 4-AP. (3) After pretreatment with 4-AP, the contraction of the control rings could significantly increased by histamine. After 4-AP treatment the Emax was significantly larger than that before 4-AP treatment. But the sensitized group rings had no such change, there was no significant difference in Emax before and after 4-AP treatment.</p><p><b>CONCLUSION</b>(1) Not K(Ca) and K(ATP) but Kv participated in regulation of the resting tone of HBSM. (2) The activity of Kv decreased in bronchial smooth muscle passively sensitized by asthmatic serum compared with that of nonsensitized group. This change might be involved in the mechanism of asthma.</p>
Subject(s)
Female , Humans , Male , Middle Aged , Asthma , Metabolism , Bronchi , Physiology , Delayed Rectifier Potassium Channels , Physiology , Immune Sera , Immunization, Passive , In Vitro Techniques , Muscle Tonus , Muscle, Smooth , PhysiologyABSTRACT
<p><b>OBJECTIVE</b>To evaluate the effect of conservative surgical management on patients with subglottic cancer.</p><p><b>METHODS</b>Nine cases with subglottic carcinoma were treated surgically from 1984 to 1999. There were T2N0 lesions in 2 cases, T3N0-1 in 3 cases and T4N0-1 in 4 cases. All the cases underwent partial laryngectomy including partial cricoid resection. Variations of a pedicled thyroid cartilage flap were used for reconstruct the cricoid defect. The pedicle based muscle was thyrohyoid, sterno-thyroid or inferior constrictor. Unilateral neck dissection was performed on 7 cases and bilateral on two.</p><p><b>RESULTS</b>The function of phonation were preserved in all cases. Eight of nine 8/9 were decanulated. Normal deglutition were achieved for all patients. The 3 and 5 year survival rates were 8/9 and 6/9, respectively.</p><p><b>CONCLUSION</b>Pedicled thyroid cartilage flap is appropriate for reconstruction of the cricoid defect in the conservative surgery of selected subglottic carcinoma.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Carcinoma, Squamous Cell , General Surgery , Cricoid Cartilage , General Surgery , Laryngeal Neoplasms , General Surgery , Laryngectomy , Plastic Surgery Procedures , Methods , Surgical Flaps , Thyroid Cartilage , TransplantationABSTRACT
<p><b>BACKGROUND</b>Potassium (K+) channels are important in regulating cell membrane potential and excitability. Although bronchial myocytes from asthmatic rats show a significant reduction in voltage-dependent delayed rectifier potassium channel (Kv) current density and higher excitability, the activity and expression of Kv in human bronchial smooth muscle cells (HBSMCs) have never been studied. The objective of this study was to investigate the effect of passive sensitization by asthmatic serum on the activity of Kv and the expression of Kv isoform Kv1.5 in HBSMCs.</p><p><b>METHODS</b>HBSMCs were randomly divided into two groups: control group (containing 10% serum from nonatopic individuals) and sensitized group (containing 10% asthmatic serum), then cultured for 24 hours. Whole-cell patch clamp, immunofluorescence staining, reverse transcription-polymerase chain reaction and Western blot techniques were used to study the effect of passive sensitization on the activity of Kv and the expression of Kv1.5 in HBSMCs.</p><p><b>RESULTS</b>The membrane potential in passively sensitized HBSMCs was significantly depolarized to -(26.7 +/- 5.2) mV compared with -(41.3 +/- 6.4) mV in the control group (P < 0.01). Passive sensitization caused a significant inhibition of Kv currents in HBSMCs, resulting in a downward shift in the current-voltage (I-V) relationship curve. At +50 mV, the peak Kv current density of passively sensitized HBSMCs was significantly decreased from (54.6 +/- 8.7) picoamperes per picofarad (pA/pF) to (32.1 +/- 7.1) pA/pF (P < 0.01). The expression level of Kv1.5 mRNA in passively sensitized HBSMCs was significantly lower than that in the control group (0.76 +/- 0.07 vs 1.04 +/- 0.13, P < 0.05). The expression of Kv1.5 protein of passively sensitized HBSMCs was also significantly reduced compared to that from the control group (984 +/- 168 vs 2200 +/- 380, P < 0.05).</p><p><b>CONCLUSIONS</b>The activity and expression of Kv were all decreased in HBSMCs passively sensitized by asthmatic serum compared with nonsensitized cells. These changes might be involved in the mechanisms of formation and development of asthma.</p>
Subject(s)
Female , Humans , Male , Asthma , Blood , Bronchi , Metabolism , Cells, Cultured , Fluorescent Antibody Technique , Immunization , Myocytes, Smooth Muscle , Metabolism , Potassium Channels, Voltage-Gated , Genetics , RNA, MessengerABSTRACT
<p><b>OBJECTIVE</b>To explore the effect of Shenmai Injection (SMI) on L-type calcium channel of diaphragmatic muscle cells in rats.</p><p><b>METHODS</b>Single diaphragmatic muscle cell of rats was obtained by the acute enzyme isolation method and the standard whole-cell patch clamp technique was used to record the inward peak L-type calcium current (IPLC) and current-voltage relationship curve of diaphragmatic muscle cells of 7 rats, and to compare the effects of SMI in various concentrations on them.</p><p><b>RESULTS</b>When keeping the electric potential at -80 mV, stimulation frequency 0.5 Hz, clamp time 300 ms, stepped voltage 10 mV, and depolarized to +60 mV, 10 microliters/ml of SMI could only cause the mean IPLC of rat's diaphragmatic muscle cells increased from -6.9 +/- 0.6 pA/pF to -7.5 +/- 0.7 pA/pF, the amplification being (9.2 +/- 2.8)%, comparison between those of pre-treatment and post-treatment showed insignificant difference. But when the concentration of SMI increased to 50 microliters/ml and 100 microliters/ml, the mean IPLC increased to -8.4 +/- 0.6 pA/pF and -9.2 +/- 0.6 pA/pF, respectively, and the amplification was (22.4 +/- 1.7)% and (34.6 +/- 4.6)% respectively, showing significant difference to that of pre-treatment (P < 0.05). However, SMI showed no significant effect on maximal activation potential and reversal potential.</p><p><b>CONCLUSION</b>SMI can activate the calcium channel of diaphragmatic muscle cells in rats, increase the influx of Ca2+, so as to strengthen the contraction of diaphragmatic muscle, which may be one of the ionic channel mechanisms of SMI in treating diaphragmatic muscle fatigue in clinical practice.</p>