Arrhythmias, Cardiac ; Brugada Syndrome ; Cardiac Conduction System Disease ; Coma ; etiology ; Electrocardiography ; Female ; Heart Conduction System ; abnormalities ; Humans ; Organophosphate Poisoning ; complications ; diagnosis

Objective: To examine expression patterns of focal adhesion kinase (FAK) and its activated form, phosphorylated FAK (pFAK),in human osteosarcoma and to investigate the correlation of FAK expression with clinicopathological parameters and prognosis. Functional consequence of manipulating FAK protein levels was also investigated in human osteosarcoma cell lines. Methods: Immunohistochemical staining was used to detect FAK and pFAK levels in pathologically archived materials from 113 patients with primary osteosarcoma. Kaplan-Meier survival and Cox regression analyses were used to evaluate prognoses. The role of FAK in cytological behavior of MG63 and 143B human osteosarcoma cell lines was studied via the FAK protein knockdown with siRNA. Cell proliferation, migration, invasiveness, and apoptosis were assessed using cell counting kit-8, Transwell, and Annexin V/PI staining methods. Results: Both FAK and pFAK were overexpressed in osteosarcoma patients. Tumor cells exhibited cytoplasmicity and occasional membranous immunoreactivity for FAK. A total of 42 cases (37.17%) mainly showed expressed pFAK in cytoplasm of osteosarcoma cells. No overexpression staining of anti-FAK and anti-pFAK antibodies was observed in normal cancellous bone tissues or negative controls. Significant differences were observed in overall survival between FAK-/pFAK- and FAK+/pFAK- groups (P=0.016), FAK+/pFAK- and FAK+/pFAK+ groups (P=0.012), and FAK-/pFAK- and FAK+/pFAK+ groups (P<0.001). All groups showed similar metastasis-free survival. Cox proportional hazard analysis showed that FAK expression profile is an independent indicator of both overall andmetastasis-free survival. siRNA-based knockdown of FAK significantly reducedmigration and invasion of MG63 and 143B cells and affected proliferation and apoptosis in osteosarcoma cells. Conclusion: Osteosarcoma malignancies in vitro and in vivo were correlated with overexpression and phosphorylation of FAK. These findings suggest that FAK plays an important biological role in osteosarcoma carcinogenesis. This study provides a better understanding of diagnostic and prognostic relevance of FAK overexpression and phosphorylation in osteosarcoma patients. Therefore, FAK and pFAK can be used as independent predictors of overall and metastasis-free survival in osteosarcoma patients.

OBJECTIVETo investigate the role and significance of P38-MAPK in the pathological process of hypoxic hypercapnia pulmonary hypertension in rats, and the protection of panax notoginoside (PNS).

METHODS(1) To set up rat pathological model of hypoxic hypercapnia pulmonary hypertension: seventy two male SD rats (200 280 g) were randomly divided into six groups (n = 12), which were normal group (N group), hypoxic hypercapnia for 3-day group (H3d), hypoxic hypercapnia for 1-week group(H1w), hypoxic hypercapnia for 2-week group (H2w), hypoxic hypercapnia for 4-week group (H4w) and PNS-injected group (Hp). The rats of PNS -injected group were injected PNS before being placed in the chamber (50 mg/(kg x d), ip), and other groups were injected normal sodium (2 ml/kg, ip). (2) The shapes of pulmonary artery were detected by HE staining. (3) Western blot was used to study the protein expression of p38-MAPK. The expression of p38-MAPK in lung tissue and pulmonary blood vessel was investigated by immunohistochemistry.

RESULTS(1) The ratio of vessel wall area/total area (WA/ TA) in H1w, H2w, H4w and Hp group was higher than that of N group (P < 0.05), but that of H3d group did not change obviously (P > 0. 05 vs N group). The ratio of WA/TA in Hp group was obviously lower than that of H4w, group (P < 0.05). (2) The levels of P-p38 protein was markedly ascended in H3d group (0.225 +/- 0.071) compared with N group (0.012 +/- 0.006), and expression of P-p38 protein was significantly positive in H1w, H2w, H4w groups. (P < 0.05). (3) As P-p38 protein in pulmonary arterial tunica intima and tunica media, sterile expression in N group (0.099 +/- 0.015) and H3d group (0.107 +/- 0.013) contrasted to H4w group (0.124 +/- 0.025, P < 0.05), then tended to rise in H2w, H4w group (P < 0.05). (4) In pulmonary tissue, the levels of P-p38 protein in PNS-injected group were lower 53.02% (P < 0.05) than those in H4w group. In pulmonary arterial tunica intima and tunica media the levels of P-p38 protein in PNS-injected group were lower 87.33% (P < 0.05) than those in H4w group.

CONCLUSIONp38-MAPK as a signal transduction may play an important role in the development of hypoxia induced pulmonary hypertension. The effect of PNS on reducing pulmonary hypertension and improving pulmonary vascular wall remodeling may be related to its inhibiting expression of p38 MAPK.

Animals ; Ginsenosides ; pharmacology ; therapeutic use ; Hypertension, Pulmonary ; drug therapy ; metabolism ; physiopathology ; Hypoxia ; metabolism ; physiopathology ; MAP Kinase Signaling System ; drug effects ; Male ; Panax notoginseng ; Phytotherapy ; Pulmonary Artery ; metabolism ; physiopathology ; Rats ; Rats, Sprague-Dawley ; p38 Mitogen-Activated Protein Kinases ; metabolism

OBJECTIVETo study the effect of extract of Ginkgo biloba (Egb761) on doxorubicin-associated cardiotoxicity in patients with breast cancer (BC).

METHODSSixty BC patients in stage IV were randomly assigned to two groups, the control group was treated with chemotherapy, using 4 cycles of PA protocol alone and the treated group with the same chemotherapy and Egb761. Changes in electrocardiogram (ECG), myocardial enzyme spectrum (MES) and ultrasono-cardiogram (USCG) before and after treatment were observed.

RESULTSAfter treatment, the incidence of abnormal ECG was lower in the treated group than in the control group (6.7% vs 30.0%); significant differences were found between the two groups in the parameters of MES (P< 0.05); USCG showed significant difference between the two groups in left ventricular diastolic diameter (LVDd), left ventricular systolic diameter (LVDs), ratio of early and late diastolic transmitral peak flow velocity (E/A) and fractional shortening (FS), while there was no significant difference in ejection fraction (EF).

CONCLUSIONEgb761 is an ideal drug for preventing and reducing the acute doxorbincin-induced cardiotoxicity; it could also be helpful for alleviating the chronic cardiotoxicity.

Adult ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Breast Neoplasms ; drug therapy ; Carcinoma, Ductal, Breast ; drug therapy ; Cardiomyopathies ; chemically induced ; physiopathology ; prevention & control ; Doxorubicin ; administration & dosage ; adverse effects ; Female ; Ginkgo biloba ; chemistry ; Humans ; Middle Aged ; Phytotherapy ; Plant Extracts ; therapeutic use ; Treatment Outcome ; Ventricular Dysfunction, Left ; physiopathology ; prevention & control

OBJECTIVETo study the effect of the small interfering RNA (siRNA)-mediated LKB1 gene silencing on the biological behavior of lung carcinoma cells.

METHODSRNA interference technique was used to silence LKB1 gene in lung carcinoma cells, and the expression of LKB1 protein were subsequently detected by Western blotting. The cell proliferation was then assessed by observing the growth curve and clone formation of the cells, and cell cycle and apoptosis were assayed by flow cytometry.

RESULTSLKB1 siRNA resulted in remarkable suppression of LKB1 expression in the lung carcinoma cells. LKB1 gene silencing resulted in accelerated cell proliferation, but cell apoptosis underwent no significant changes.

CONCLUSIONDown-regulation of LKB1 gene expression can stimulate malignant biological behavior of lung carcinoma cells.

Animals ; Apoptosis ; genetics ; Blotting, Western ; Cell Line, Tumor ; Cell Proliferation ; Flow Cytometry ; Gene Silencing ; Humans ; Lung Neoplasms ; genetics ; pathology ; Protein-Serine-Threonine Kinases ; deficiency ; genetics ; RNA, Small Interfering ; genetics

OBJECTIVETo investigate the relationship between blood pressure variability (BPV) and left ventricular diastolic function in patients with essential hypertension.

METHODSLeft ventricular diastolic function of 252 hypertensive patients were assessed by early (E) diastolic transmitral flows to early diastolic mitral annular velocity (Ea) (E/Ea) ratio derived from Doppler echocardiography. Patients were divided into two groups according to normal left ventricular diastolic function group (E/Ea<15, n = 168) and left ventricular diastolic dysfunction group (E/Ea ≥ 15, n = 84). All patients were monitored by ambulatory blood pressure. Standard deviation (SD) and coefficient of variation (CV) of blood pressure were calculated as the BPV. Relationship between BPV and left ventricular diastolic function were analyzed by multivariate logistic regression analysis.

RESULTSAll-day average diastolic blood pressure(DBP), the day systolic blood pressure (SBP), night SBP, night DBP, SBPSD, DBPSD and DBPCV in the left ventricular diastolic dysfunction group were significantly higher than in the normal diastolic function group (all P < 0.05). Multivariate logistic regression analysis showed that left ventricular diastolic dysfunction was associated with SBPSD (OR:1.126, 95%CI:1.054-1.203, P < 0.01), SBPCV (OR:1.127, 95%CI:1.036-1.225, P < 0.01) in this patient cohort.

CONCLUSIONHigh variability of SBP is correlated with left ventricular diastolic dysfunction in hypertensive patients.

Adult ; Aged ; Blood Pressure ; physiology ; Diastole ; physiology ; Essential Hypertension ; Female ; Humans ; Hypertension ; physiopathology ; Logistic Models ; Male ; Middle Aged ; Ventricular Function, Left ; physiology

OBJECTIVETo evaluate the efficacy and safety of a once daily valsartan/amlodipine 80/5 mg combination tablet in Chinese mild to moderate hypertensive patients without adequate blood pressure control by monotherapy.

METHODSTwo multicenter, randomized, double-blind, double dummy, active-controlled, parallel group trials were conducted. After a washout period (no medication) of 1-4 weeks, patients with Mean Sitting Diastolic Blood Pressure (MSDBP) > or = 95 mm Hg (1 mm Hg = 0.133 kPa) and < 110 mm Hg received a monotherapy of either Amlodipine 5 mg (in study 1) or valsartan 80 mg (in study 2) for 4 weeks. Patients with MSDBP > or = 90 mm Hg and < 110 mm Hg at the end of the monotherapy period were randomized to receive valsartan/amlodipine 80/5 mg treatment, or continue with the monotherapy.

RESULTSIn study 1, compared with amlodipine 5 mg, valsartan/amlodipine 80/5 mg once daily further reduced mean sitting systolic blood pressure (MSSBP)/MSDBP 4.4/3 mm Hg (P < 0.0001). In study 2, compared with valsartan 80 mg, valsartan/amlodipine 80/5 mg once daily further reduced MSSBP/MSDBP 6.4/4.2 mm Hg (P < 0.0001). The blood pressure (BP) control rates (BP < 140/90 mm Hg) of combination treatment group were 71.0% and 71.2% respectively, and significantly higher than the monotherapy groups in both trials. Incidence of adverse events was comparable in monotherapy and combination therapy groups.

CONCLUSIONOur results showed that valsartan/amlodipine 80/5 mg was superior to amlodipine 5 mg or valsartan 80 mg alone in lowering blood pressure and BP control in patients with mild to moderate hypertension not adequately controlled with amlodipine 5 mg or valsartan 80 mg monotherapy. No new or unexpected safety issues were identified with valsartan/amlodipine combination therapy compared with monotherapy.

Adult ; Amlodipine ; administration & dosage ; Antihypertensive Agents ; administration & dosage ; Blood Pressure ; Double-Blind Method ; Drug Therapy, Combination ; Female ; Humans ; Hypertension ; drug therapy ; Male ; Middle Aged ; Tetrazoles ; administration & dosage ; Valine ; administration & dosage ; analogs & derivatives ; Valsartan

Objective To observe the expression of stromal cell-derived factor-1α(SDF-1α)and glial cell line-derived neurotrophic factor(GDNF)genes in bone marrow stromal cells(BMSCs)of rhesus monkey. Methods With gene recombination technique,SDF-1α and GDNF genes obtained from cDNA were subcloned into pBudce4.1 vector to get pBudCE4.1-SDF-1-GDNF, evaluated by restriction enzyme analysis and sequencing analysis.The pBudCE4.1-SDF-1α-GDNF was transfected into BMSCs with lipofectamine2000.After 48 h,the expression of SDF-1α and GDNF was measured by RT-PCIL,Western blotting and immunohistochemistry. Results Correct construction of pBudCE4.1-SDF-1α-GDNF was identified by enzyme restriction analysis and sequencing analysis.Western blotting and immunohistochemistry confirmed the expressions Of SDF-1α and GDNF genes in the transfected cells.The protein expressions of GDNF and SDF-1α in the GDNF and SDF-1α transfected BMSCs were 6 times higher than those in the negative control cells. Conclusion The pBudCE4.1-SDF-1α-GDNF can successfully express SDF-1α and GDNF in BMSCs of rhesus monkey,which may be used in gene therapy for Parkinson's disease.

BACKGROUNDWith economic growth and urbanization there have been significant changes in the life style and diet of urban residents in large cities of China, which is experiencing a rapid increase in the prevalence of diabetes. While high prevalence of diabetes has been reported, little is known of the long-term effects of diabetes in such a large population. The aim of this study was to estimate the morbidity rate of diabetic peripheral neuropathy (DPN) in a Chinese urban diabetic population with more than 10 years' disease duration, and evaluate the relevant risk factors. The clinical manifestation of DPN and pain status was also assessed.

METHODSFive hundred and sixty-five diabetes patients were recruited into the study. Symptoms and examination helped diagnose neuropathy. The clinical manifestation of DPN was assessed with a visual analog pain score (VAS). Diabetic complication status was determined from medical records. Serum lipids and lipoproteins, glycosylated hemoglobin (HbA1c), and the urinary albumin excretion rate were measured.

RESULTSThe morbidity rate of DPN was 46.6%. HbA1c, hyperlipidemia, and retinopathy were significantly associated with neuropathy, and these risk factors were correlated with other diabetic micro and/or macrovascular complications. The average VAS pain score of the DPN patients was 4.12 ± 2.07. Severe and moderate pain was experienced by 11.4% and 40.5% respectively of DPN patients. About 3.7% of diabetic subjects had lower limb ulcer or amputation.

CONCLUSIONSThe morbidity rate of DPN for diabetic patients with > 10 years duration is very high compared to the range reported for other populations in the world. The risk factors for DPN include HbA1c, hyperlipidemia, and retinopathy. In long-standing diabetic patients, DPN was not associated with diabetic duration, and half of the DPN patients experienced considerable daily suffering.

Aged ; China ; Diabetic Neuropathies ; epidemiology ; metabolism ; physiopathology ; Female ; Glycated Hemoglobin A ; metabolism ; Humans ; Hyperlipidemias ; epidemiology ; metabolism ; physiopathology ; Male ; Middle Aged ; Pain ; etiology ; Risk Factors ; Urban Population

OBJECTIVETo construct a eukaryotic fluorescent expression vector of truncated lymphoid enhancer-binding factor 1 (LEF-1) gene and investigate its effect on the proliferation and apoptosis of human colonic carcinoma cell line SW480.

METHODSTruncated LEF-1 gene was obtained by PCR and DNA recombination from human lymphoid node cDNA library. The PCR product of LEF-1 gene was inserted into the plasmid pMD-18T and sequenced. The truncated LEF-1 gene was inserted into the eukaryotic expression plasmid pcDNA3.1 and fused with mRFP for tracing. Using Lipofectamine 2000, the plasmid pcDNA3.1-LEF-1-mRFP was transfected into Hela cells and detected by Western blotting and fluorescence activated cell sorting (FACS). The changes in the growth, proliferation and apoptosis of the SW480 cells were observed after transfection with the plasmids.

RESULTSThe truncated LEF-1 gene was successfully cloned. After transfection with the plasmid pcDNA3.1-LEF-1-mRFP, the Hela cells expressed the product of LEF-1 as detected by Western blotting and FACS. The growth and proliferation of SW480 cells was inhibited and the cell apoptosis increased after transfection with the plasmid pcDNA3.1-LEF-1-mRFP, which also caused cell cycle arrest in G0/1 phase.

CONCLUSIONThe eukaryotic expression fluorescent vector pcDNA3.1-LEF-1-mRFP has been constructed and expressed in eukaryotic cell line successfully. The truncated LEF-1 protein expressed in the transfected SW480 cells results in inhibition of the cell growth and proliferation with increased cell apoptosis and cell cycle arrest in G0/1 phase.

Cell Line, Tumor ; Colonic Neoplasms ; metabolism ; pathology ; Fluorescence ; Genetic Vectors ; genetics ; Humans ; Lymphoid Enhancer-Binding Factor 1 ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics