[Abstract] Objective To explore the relationship between Kruppel-like factor 4 (KLF-4) and E-cadherin in human peritoneal mesothelial cells (HPMCs), and the expression and function of KLF-4 in the animal model of peritoneal fibrosis induced by high glucose peritoneal dialysate. Methods Co-transfection in HPMCs with the plasmid of KLF-4 and the bind site or mutant in the promoter region of E-cadherin, and then the luciferase activity was measured of the each bind site and its matched mutants to estimate whether KLF-4 can combine with the bind site in the promoter region of E-cadherin; Chromatin immunocoprecipitation (CHIP) was exploited to verify if KLF-4 can combine with the bind site in the promoter region of E-cadherin; Real-time PCR and Western blotting were performed to detect the expression of E-cadherin at the bind site and matched mutants of b, d, f and g. Thirty SD rats were randomly divided into saline group, peritoneal dialysate group and experimental group (10 each). Rats in saline group were given intraperitoneal injection with 0.9% NaCl, in peritoneal dialysate group were given with 4.25% high glucose peritoneal dialysate, and in experimental group were given via tail vein with 4.25% high glucose peritoneal dialysate and the mixture of KLF-4 plasmid suspension containing ultrasound microbubble. To observe the peritoneal tissue thickness of the 3 groups of rats by Hematoxylin and Eosin staining. Masson trichrome staining was performed to detect the deposition of collagen fibers in peritoneal tissue of the 3 groups of rats. Immunohistochemistry was used to detect the expression level of KLF-4, E-cadherin, α-SMA and fibronectin (FN) in peritoneal tissue of the 3 groups of rats. Results Promoter luciferase reporter gene and CHIP results showed that KLF-4 can combine with the bind site in the promoter region of E-cadherin in HPMCs. Real-time PCR and Western blotting showed that KLF-4 can positively regulate the expression of E-cadherin. HE staining showed that the peritoneal tissue was obviously thickened in rats of peritoneal dialysate group [(105.91±12.0) μm] than in rats of saline group [(20.89±5.39) μm] and of experimental group [(23.05±6.07) μm] with statistical significance (P<0.05), but the difference between saline group and experimental group was not statistically significant (P>0.05). Masson staining showed that the deposition of collagen fiber significantly increased in peritoneal dialysate group (0.89±0.09) than in saline group (0.19±0.03) and experimental group (0.15±0.06) with statistical significance (P<0.05), but the difference between saline group and experimental group was not statistically significant (P>0.05). Immunohistochemistry results showed that the expressions of KLF-4 and E-cadherin were obviously lower in peritoneal dialysate group (0.27±0.09, 0.31±0.03) than in saline group (0.79±0.19, 0.83±0.13) and experimental group (0.85±0.11, 0.76±0.11) with statistically significant difference (P<0.05), but no significant difference existed between saline group and experimental group (P>0.05). In contrast, the expressions of α-SMA and FN were evidently higher in peritoneal dialysate group (0.83±0.09, 0.63±0.09) than in saline group (0.22±0.08, 0.30±0.07) and experimental group (0.19±0.05, 0.11±0.03) with statistically significant difference (P<0.05), but no evident difference existed between saline group and experimental group (P>0.05). Conclusion KLF-4 may positive regulate the expression of E-cadherin by combining with the bind site in the promoter region of E-cadherin, and inhibit the peritoneal fibrosis induced via high glucose peritoneal dialysate.

Objective To explore the role of Kruppel-like factor 4 (KLF-4) in phenotypic transition of human peritoneal mesothelial cells (HPMCs) induced via high glucose. Methods HPMCs were induced by 50mmol/L glucose for 72 hours, the expressions of epithelium-cadherin (E-cadherin), KLF-4, α-smooth muscle actin (-SMA), connective tissue growth factor (CTGF) and miRNA-143 were detected by Real-time PCR and Western blotting, respectively. The treated cells were transfected with LVKLF-4 and inhibitor, the untreated cells were transfected with shRNA-KLF-4 and mimic. The mRNA and protein expressions of KLF-4, E-cadherin, α-SMA, CTGF and miRNA-143 were detected by Real-time PCR and Western blotting, respectively. Results Real-time PCR showed that the expression of E-cadherin decreased and of α-SMA, CTGF and miRNA-143 increased, but of KLF-4 not changed in high glucose treated cells. Western blotting showed that the expression of KLF-4 and E-cadherin decreased. Upregulating KLF-4 increased the expression of E-cadherin, but decreased the expression of α-SMA and CTGF. Down-regulating KLF-4 decreased the expression of E-cadherin, but augment the expression of α-SMA and CTGF. Conclusion High glucose may induce the down-regulation of KLF-4 protein, and SRF- miRNA-143-KLF-4 signal pathway axis may be involved in the process of HPMC phenotypic transition.

Objective To study the morbidity of cough variant asthma (CVA) among patients with chronic cough syndrome and its relative risk factors. Methods Patients were recruited with detailed history on their illness. Data were collected on physical examination, chest X-ray, eosinophil cell counts, pulmonary ventilation with histamine stimulating test and bronchi dilation test. According to available data, diagnosis of CVA was confirmed and the relative factors Questionnaire form was completed for each patient. Results Among 473 patients with chronic cough, 95 (44 male and 51 female) were confirmed to be CVA (20.08%). Analysis of the relative factors suggested that CVA was associated with multiple factors. Morbidity of CVA was associated with season, personal histories on allergy and family history on asthma, CVA could be induced by upper respiratory tract infection, inhale of oil vapor, acrimony air, over-burdened physical exercises etc. Conclusion For patients with chronic cough symptom, clear diagnosis of CVA, avoid of passable risk factors and timely medical intervention when necessary, would be helpful in controlling clinical courses and improving the prognosis of the disease.

BACKGROUNDThere is no research, either at home or abroad, focusing on assessing the cardiopulmonary functional reserve and exercise tolerance in patients with pulmonary embolism (PE), but the benefits of early exercise are well recognized. The goals of this study were to assess cardiopulmonary functional reserve in treated PE patients using the inert gas rebreathing method of the cardiopulmonary exercise test (CPET), and to compare it with traditional methods.

METHODSCPET on the bicycle ergometer were performed in 40 patients with age, gender, body mass index, systolic blood pressure, and pulmonary function matched. The first group was the PE group composed of 16 PE patients (5 male, 11 female) who were given the standard antithrombotic therapy for two weeks. The second group was composed of 24 normal individuals (10 male, 14 female). Both groups were evaluated by cardiac ultrasound examination, 6-minute walking test (6MWT), and CPET.

RESULTS(1) Right ventricular systolic pressure (RVSP) in the PE group increased significantly compared to the control group, (34.81 ± 8.15) mmHg to (19.75 ± 3.47) mmHg (P < 0.01). But neither right atrial end-systolic diameter (RASD) nor right ventricular end-diastolic diameter (RVDD) in the PE patients had changed when compared with the controls. The 6-minute walk distance was significantly reduced in the PE patients compared with normal subjects, (447.81 ± 79.20) m vs. (513.75 ± 31.45) m (P < 0.01). Both anaerobic threshold oxygen consumption (VO(2)AT) and peak oxygen consumption (VO(2)peak) were significantly lower in patients with PE, while CO(2) equivalent ventilation (VE/VCO(2) slope) was higher; VO(2)AT (9.44 ± 3.82) ml×kg(-1)×min(-1) vs. (14.62 ± 2.93) ml×kg(-1)×min(-1) (P < 0.01) and VO2peak (12.26 ± 4.06) ml×kg(-1)×min(-1) vs. (23.46 ± 6.15) ml×kg(-1)×min(-1) (P < 0.01) and VE/VCO(2) slope 35.47 ± 6.66 vs. 26.94 ± 3.16 (P < 0.01). There was no significant difference in resting cardiac output (CO) between the PE and normal groups, whereas peak cardiac output (peak CO) and the difference between exercise and resting cardiac output (ΔCO) were both significantly reduced in the PE group; peak CO (5.97 ± 2.25) L/min to (8.50 ± 3.13) L/min (P < 0.01), ΔCO (1.29 ± 1.59) L/min to (3.97 ± 2.02) L/min (P < 0.01). (2) The 6-minute walk distance did not correlated with CPET except for the VO2 peak in patients with PE, r = 0.675 (P < 0.01).

CONCLUSIONSThe cardiopulmonary functional reserve was reduced in patients with PE. CPET is an accurate, quantitative evaluation of cardiopulmonary functional reserve for PE patients.

Aged ; Exercise Test ; methods ; Exercise Tolerance ; physiology ; Female ; Humans ; Male ; Middle Aged ; Oxygen Consumption ; physiology ; Pulmonary Embolism ; physiopathology ; therapy

BACKGROUNDThe transradial approach is regarded as a useful vascular site for coronary procedures. The aim of this study was to test whether 4Fr catheters assisted by ACIST variable rate injector system can produce comparable angiographic quality and reduce the risk of radial artery injury compared to hand manifold 6 Fr catheters.

METHODSA total of 1816 patients were studied consecutively, among whom 856 patients received coronary angiography by 4 Fr catheters (4Fr group) and 960 patients by 6 Fr catheters (6Fr group). Angiographic and procedural characteristics were observed and recorded. The luminal inner radial arterial diameter before and after the procedure were collected.

RESULTSThe baseline clinical characteristics were similar in both groups. There were no significant differences in procedure time, radiation dose and quality scores in both groups (P > 0.05), but more contrast media was delivered in the 6Fr group (P < 0.001). The mean radial arterial diameter six months after the procedure in the 6Fr group reduced significantly compared to that measured one day prior to the procedure (P < 0.001).

CONCLUSIONSCoronary angiography using the 4Fr catheters with Acist power injection system can achieve an acceptable diagnostic quality while at the same time minimizing radial artery injury and contrast media consumption.

Aged ; Cardiac Catheterization ; instrumentation ; Coronary Angiography ; instrumentation ; methods ; Female ; Humans ; Male ; Middle Aged ; Radial Artery ; diagnostic imaging ; Ultrasonography

Objective To explore the effect of long noncoding RNA-ATB (LncRNA-ATB) on phenotypic transition and proliferation of human peritoneal mesothelial cells (HPMCs) induced by high glucose.Methods HPMCs used in experiment were divided into three groups:control group,mannitol group and hypertonic glucose group.HPMCs in control group received no treatment,and in hypertonic glucose group and mannitol group were treated with 50mmol/L D-glucose and isotonic mannitol for 72 hours,respectively.Real-time PCR was employed to detect the mRNA expression of LncRNA-ATB,E-cadherin,α-smooth muscle actin (α-SMA),connective tissue growth factor (CTGF),Cyclin D1,cyclin dependent kinase inhibitor 4 (CDK4),protein 27 (p27)and proliferating cell nuclear antigen (PCNA).Western blotting was performed to detect the proteins expression of E-cadherin,α-SMA,CTGF,Cyclin D1,CDK4,p27 and PCNA,and flow cytometry was used to test the cell cycle.Lentivirus artifice was used to up-or down-regulate the expression of LncRNA-ATB in untreated HPMCs.Real-time PCR was employed to detect the mRNA expression of E-cadherin,α-SMA and CTGF,Western blotting was performed to detect the proteins expression of E-cadherin,α-SMA and CTGF,and flow cytometry was used to test the cell cycle.Results It is revealed by Real-time PCR,Western blotting and flow cytometry that the expressions increased of LncRNA-ATB,α-SMA,CTGF,Cyclin D1,CDK4 and PCNA induced by hypertonic glucose,and decreased of E-cadherin and p27 (P＜0.05).Up-regulation of LncRNA-ATB promoted HPMCs phenotypic transition and proliferation,while down-regulation alleviated HPMCs phenotypic transition and proliferation.Conclusion Hypertonic glucose may accelerate HPMCs phenotypic transition and proliferation by up-regulating the expression of LncRNA-ATB.

Objective To explore the effects of p53 up-regulated modulator of apoptosis (PUMA)-α protein on the apoptosis and fibrosis of human peritoneal mesothelial cells (HPMCs) induced by high glucose.Methods HPMCs were induced by 50mmol/L D type glucose or mannitol for 72 hours respectively,flow cytometry was employed to detect the rate of apoptotic cells,and theexpression levels of apoptosis-and fibrosis-related proteins were detected by Western blotting.The untreated HPMCs were transfected with Lenti-PUMA-α,and the treated cells were transfected with shRNA-PUMA-α,the number of apoptotic cells and the expression levels of apoptosis-and fibrosis-related proteins were detected with the methods mentioned above.Results Flow cytometry showed that the rate of apoptotic HPMCs increased after being induced by high glucose for 72 hours,and Western blotting revealed that the expression levels of pro-apoptotic and pro-fibrotic related proteins increased,but the arrestins of apoptosis and fibrosis-related proteins decreased.Up-regulation of PUMA-α promoted apoptosis and fibrosis,while down-regulation of PUMA-α alleviated apoptosis and fibrosis of HPMCs.Conclusion High glucose may accelerate apoptosis and fibrosis of HPMCs by up-regulating the expression of PUMA-α.

OBJECTIVETo investigate the expression of heparin-binding epidermal growth factor-like growth factor (HB-EGF) in patients with gastric carcinoma in different stages.

METHODSThe expressions of HB-EGF protein and mRNA in normal gastric tissues, metaplasic intestinal mucosa, early-stage gastric cancer and advanced-stage gastric cancer tissues were detected by immunohistochemistry and in situ hybridization.

RESULTSHB-EGF expression was only detected in the parietal cells of the gastric fundic glands and in gastrin cells of the pyloric glands in normal gastric tissues. Weak HB-EGF expression was detected in the epithelial cells of the gastric mucosa in intestinal metaplasic mucosa, and the expression increased in all layers of the gastric mucosa in early-stage gastric cancer. Intense HB-EGF expression was observed in advanced gastric cancer.

CONCLUSIONIncreased HB-EGF expression may be implicated in the pathogenesis and development of gastric carcinoma.

Adenocarcinoma ; metabolism ; pathology ; Adult ; Aged ; Female ; Gastric Mucosa ; metabolism ; Heparin-binding EGF-like Growth Factor ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; metabolism ; Male ; Middle Aged ; RNA, Messenger ; genetics ; metabolism ; Stomach Neoplasms ; metabolism ; pathology ; Tumor Cells, Cultured

Objective To investigate the effects of peroxisome proliferator-activated receptor (PPAR) α/γ agonist on atherosclerotic plaque stabilization in diabetic LDL receptor knockout(LDLr-/-)mice.Methods Female 4-week-old LDLr-/-mice fed with high-glucose and high-fat diet for 4 weeks were randomly divided into three groups (n =15 each): control group (only fed with high-glucose and high-fat diet),diabetic group [induced by high-glucose and high-fat diet combined with a low-dose of streptozotocin (STZ)] without tesaglitazar and with tesaglitazar (20 μg/kg oral treatment).Mter 6 weeks,the mice were sacrificed,body weight,fasting blood glucose (Glu),total cholesterol (TC),triglyceride (TG) levels were measured.The expression of ICAM-1,VCAM-1,MCP-1 in the brachiocephalic atheroselerotic lesions were determined by Western blot and immunohistochemistry,respectively.Brachiocephalic artery was prepared for morphologic study (HE,oil red O,Sirius red staining) and immunohistochemical analysis (macrophage surface molecule-3,α-smooth muscle actin),respectively.Results Serum TC [(32.34 ± 3.26) mmol/Lvs.(16.17 ±1.91) mmol/L],TG[(3.57 ±0.99) mmol/L vs.(2.21 ±0.11) mmol/L] and Glu[(15.21 ±4.67) mmol/L vs.(6.89 ± 0.83)mmol/L] levels were significantly higher in diabetic group than in the control group (all P ＜ 0.01).The expression of ICAM-1 (2.31 ± 0.35 vs.1.34 ± 0.21),VCAM-1 (1.65 ±0.14 vs.0.82 ±0.26),MCP-1 (2.27 ± 0.16 vs.1.56 ± 0.23) were significantly upregulated in diabetic group compared with control group (all P ＜ 0.01).Brachiocephalic atheroselerotic plaque area [(4.597 ±1.260) ×103 μm2 vs.(0.075 ±0.030) ×103 μm2],lipid deposition [(47.23 ±2.64)％ vs.(9.67 ±1.75)％],Mac-3 positive area [(19.15 ± 3.51)％ vs.(1.72 ± 0.16)％],α-smooth muscle actin [(5.54 ± 1.17) ％ vs.(2.13 ± 0.41) ％] and collagen content [(4.27 ± 0.74) ％ vs.(0.43 ± 0.09) ％]were all significantly larger/higher in diabetic LDLr-/-mice than in the control group (all P ＜ 0.01).While tesaglitazar treatment significantly reduced serum TC [(30.47 ± 3.18) mmol/L],TG [(3.14 ± 0.71)mmol/L] and Glu [(7.92 ± 1.28) mmol/L] levels (all P ＜ 0.01).Similarly,the expression of ICAM-1 [(1.84 ± 0.22)],VCAM-1 [(1.27 ± 0.11)],MCP-1 [(1.83 ± 0.24)],brachiocephalic atheroselerotic lesion area [(1.283 ± 0.410) × 103 μm2],lipid deposition [(23.52 ± 1.39) ％] were also significantly reduced by tesaglitazar (all P ＜ 0.05).Moreover,tesaglitazar increased o-smooth muscle actin [(9.46 ±1.47) ％] and collagen content [(6.32 ± 1.15) ％] in diabetic LDLr-/-mice (all P ＜ 0.05).In addition,lipid deposition and Mac-3 positive areas [(10.67 ± 0.88)％ vs.(15.83 ± 1.01)％] in the aortic root were also reduced in tesaglitazar treated diabetic LDLr-/-mice (P ＜ 0.01).Conclusions Tesaglitazar has antiinflammatory effects in the diabetic LDLr-/-mice.Tesaglitazar could reduce lipid deposition,increase collagen and α-SMA content in the brachiocephalic atheroselerotic lesions,thus,stabilize atherosclerotic plaque in this model.

Objective To observe the effects of aerobic exercise on cardiac output during exercise in patients with chronic heart failure (CHF). Methods A total of 50 CHF patients ( echocardiography measured left ventricular ejection fraction ＜ 0. 49) were enrolled in the study and randomly divided into aerobic exercise group ( n = 25 ) and control group ( n = 25 ). Cardiopulmonary exercise testing (CPET) was performed. Patients of aerobic exercise group underwent aerobic exercise according to aerobic exercise prescription and exercise intensity is decided by anaerobic threshold before 10 J/s ( 1 minute before) of the oxygen consumption. After 6 supervised aerobic exercise training sessions in the hospital, patients were asked to perform the home-based aerobic exercise training. Patients in control group were required to maintain daily physical activities. CPET were reviewed 3 months later. Results Cardiac output(CO) ,peak CO, peak cardiac power output ( peak CPO), resting heart rate ( HR), heart rate at AT ( HR AT), HR peak, resting mean arterial pressure (MAP), peak MAP at baseline were similar between aerobic exercise group and control [(4. 2 ± 2. 0) L/min vs. (3. 3 ± 1.0) L/min, (6. 2 ± 2. 7 ) L/min vs. (5. 2 ± 1.8 ) L/min,( 1.8 ± 2. 9) L/min vs. (2. 0 ± 1.8 ) L/min, ( 1.3 ± 0. 5 ) J/s vs. ( 1.2 ± 0. 5 ) J/s, (76. 8 ± 13.5 ) beats/min vs. (73.4 ± 11.9 ) beats/min, ( 91.5 ± 11.3 ) beats/min vs. ( 92. 6 ± 12. 4 ) beats/min, ( 106. 0 ± 12. 9 )beats/min vs. ( 108. 3 ± 17.4) beats/min, (80. 8 ±9. 9)mm Hg( 1 mm Hg =0. 133 kPa) vs. (87. 6 ± 13.3)mm Hg, ( 98. 8 ± 12. 4 ) mm Hg vs. ( 102. 7 ± 13.9 ) mm Hg, all P ＞ 0. 05]. Compared to baseline, CO, peak CO, peak CPO, HR, HR AT, HR peak, MAP, peak MAP after 3 months were similar between aerobic exercise group and control( all P ＞ 0. 05 ). The differences between baseline and 3 months later expressed as △CO,△peak CO,△peak CPO, △HR, △HR AT, △HR peak, △MAP, △peak MAP were also similar between aerobic exercise group and control group [( - 0. 7 ± 2. 4) L/min vs. (0. 7 ± 2.0) L/min, ( 1.1 ± 2. 6 ) L/min vs.(1.4±2. 1)L/min,(0. 1 ±3.7)L/min vs. ( -0.2 ±2. 5) L/min, (0. 2 ± 1.0) J/s vs. (0.2 ±0.5)J/s,( - 0. 4 ± 7. 6) beats/min vs. ( 1.9 ± 9.9 ) beats/min, ( 3.4 ± 11.3 ) beats/min vs. ( - 2. 8 ± 7.6 )beats/min, (8. 9 ± 14. 5 )beats/min vs. (3.7 ± 14. 4)beats/min, (1.5 ± 12. 8 )mm Hg vs. ( - 1.3 ± 11.1 )mmHg,(6.4±18. 9)mm Hg vs. (1.3±12.3)mm Hg,all P＞0. 05]. Conclusion Three months aerobic exercise training did not improve cardiac output and related parameters during exercise in this cohort patients with CHF.