Objective To investigate the molecular mechanisms for the development of cardiac hypertrophy in hypertension,the present study provided the differential protein expression analysis of hypertrophied heart at differ- ent stages in spontaneously hypertensive rats (SHR).Methods The profiles of protein expression of left ventricu lar myocardium in SHR and its normotensive control Wistar-Kyoto (WKY) rats at the age of 4 and 20 weeks were analyzed with two-dimensional gel electrophoresis (2-DE) in combination with matrix assisted laser desorption ioniza- tion-time of flight (MALDI-TOF-TOF) mass spectrometry.Results Although the blood pressure of SHR was normal at 4 weeks age,hypertrophy of the left ventricle had already developed.The expression pattern in the hy- pertrophic myocardium was found 27 modulated proteins,20 of which were identified.These proteins are involved in reactions of energy metabolism,mitoehondrial oxidative phosphorylation and oxidative stress,etc.The expres- sion of 13 proteins was significantly changed in SHR rats at early stage prior to the development of sustained hyper- tension,while the expression changes of other 7 proteins occurred only at late stage in SHR rats when the blood pressure was significantly elevated.Conclusions lncrease in glycolysis and decrease in oxidation of fatty acid and glucose was shown in the hypertrophied myocardium from early stage in SHR prior to the development of hyperten sion.The significant changes in protein expression of the mitochondrial electron transport chain and antioxidative molecules support the hypothesis that oxidative stress promotes and accelerates the development of hypertensive car- diac hypertrophy.

BACKGROUNDCathepsin S and its endogenous inhibitor cystatin C are implicated in the pathogenesis of atherosclerosis, especially in the plaque destabilization and rupture leading to acute coronary syndrome. However, whether circulating cathepsin S and cystatin C also change in association with coronary plaque morphology is unknown yet.

METHODSWe recruited 98 patients with unstable angina (UA, n = 6) or stable angina (SA, n = 2) who had a segmental stenosis resulting in > 20% and < 70% diameter reduction in one major coronary artery on coronary angiography. Thirty-one healthy subjects served as controls. Intravascular ultrasound (IVUS) was used to evaluate plaque morphology. Plasma cathepsin S and cystatin C were measured as well.

RESULTSAt the culprit lesion site, plaque area ((7.85 +/- 2.83) mm(2) vs (6.53 +/- 2.92) mm(2), P = 0.027), plaque burden ((60.92 +/- 11.04)% vs (53.87 +/- 17.52)%, P = 0.025), remodeling index (0.93 +/- 0.16 vs 0.86 +/- 0.10, P = 0.004) and eccentricity index (0.74 +/- 0.17 vs 0.66 +/- 0.21, P = 0.038) were bigger in UA group than in SA group. Plasma cathepsin S and cystatin C were significantly higher in patients than in controls (P < 0.01). Plasma cathepsin S was higher in UA group ((0.411 +/- 0.121) nmol/L) than in SA group ((0.355 +/- 0.099) nmol/L, P = 0.007), so did the plasma cystatin C ((0.95 +/- 0.23) mg/L in UA group, (0.84 +/- 0.22) mg/L in SA group; P = 0.009). Plasma cathepsin S positively correlated with remodeling index (r = 0.402, P = 0.002) and eccentricity index (r = 0.441, P = 0.001), and plasma cystatin C positively correlated with plaque area (r = 0.467, P < 0.001) and plaque burden (r = 0.395, P = 0.003) in UA group but not in SA group.

CONCLUSIONSPlasma cathepsin S and cystatin C increased significantly in UA patients. In angina patients, higher plasma cathepsin S may suggest the presence of vulnerable plaque, and higher plasma cystatin C may be a clue for larger atherosclerotic coronary plaque.

Adult ; Aged ; Aged, 80 and over ; Cathepsins ; blood ; Coronary Artery Disease ; blood ; diagnostic imaging ; pathology ; Cystatin C ; blood ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Ultrasonography, Interventional ; methods

OBJECTIVETo investigate whether S. aureus could activate NF-κB signaling pathway in human osteoblasts.

METHODSImmunoblot and electrophoretic mobility shift assay were used to detect the degradation of I-κBα and activation of NF-κB in human osteoblasts following infection with S.aureus, respectively, and there were investigated the activated state of NF-κB signaling pathway in human osteoblasts. In addition, enzyme-linked immunosorbent assay was used to measure the secretion of IL-6 in culture supernatants, which was represented as one of important cytokines in osteomyelitis, and an inhibitor of NF-κB, SN50, which was added to human osteoblasts culture prior to 1 hour at 50 µmol/L before the infection of S.aureus, was used to determine whether S.aureus-activated NF-κB signaling pathway regulates IL-6 secretion of human osteoblasts.

RESULTSS.aureus could induce the degradation of I-κBα (I-κBα(15 min)/I-κBα(0 min) = 0.409 ± 0.245 and I-κBα(30 min)/I-κBα(0 min) = 0.061 ± 0.010) and activation of NF-κB in human osteoblasts in a time and dose-dependent manner following infection. In addition, the secretion of IL-6 in the supernatants of human osteoblasts ((2.17 ± 0.11) µg/L) was suppressed by 50 µmol/L SN50 compared to without the addition of SN50 ((3.58 ± 0.31) µg/L) (F = 174.25, P < 0.05).

CONCLUSIONSS.aureus could activate NF-κB signaling pathway in human osteoblasts, which could regulate cytokines secretions of human osteoblasts.

Cells, Cultured ; Humans ; Interleukin-6 ; secretion ; NF-kappa B ; metabolism ; Osteoblasts ; metabolism ; Signal Transduction ; Staphylococcal Infections ; metabolism

BACKGROUNDDevelopment of vulnerable lesions is not limited to the target lesions, but a pan-coronary process. Such lesions are identified by positive remodeling (intravascular ultrasound (IVUS) and complex lesions (angiography)). The prevalence of lesions with vulnerable characteristics in patients with stable angina was not well known. The purpose of the present study was to evaluate the relationship between coronary artery remodeling and incidence of angiographic complex lesions and its calcification in stable angina patients.

METHODSOne hundred and sixty-one stable angina patients (95 males, aged (68 +/- 11) years) with 161 de novo target lesions were studied using pre-interventional IVUS. Remodeling index was defined as the lesion divided by reference vessel area; positive remodeling was defined as remodeling index > 1.05. Besides the 161 target lesions, there were 613 angiographic lesions with > 30% diameter stenoses, classified as complex or smooth. Multiple complexes were defined as more than one complex lesion in one patient. Stenoses of at least 70% were described as tight. Calcium arc area was used as a new method to quantify coronary calcification.

RESULTSFifty-six patients had positive remodeling target lesion, while 105 did not. The overall number of lesions with a diameter stenoses > 30% was similar in patients with or without positive remodeling, and the frequency of angiographically complex lesions was higher in positive remodeling patients, especially at non-target site. Calcium arc area was smaller in patients with positive remodeling.

CONCLUSIONSPositive remodeling on intravascular ultrasound was associated with more complex lesions angiographic findings, especially at non target site. Positive remodeling was found less calcified in patients with stable angina.

Aged ; Aged, 80 and over ; Angina Pectoris ; diagnostic imaging ; pathology ; Coronary Angiography ; Coronary Artery Disease ; diagnostic imaging ; pathology ; Coronary Vessels ; pathology ; Female ; Humans ; Male ; Middle Aged ; Ventricular Remodeling ; physiology

OBJECTIVETo investigate the expression and significance of Cyclin D1 in oral squamous cell ma (OSCC).

METHODSA immunohistochemistry method, Envosion, was employed to test the manifesting Cyclin D1 in pathological slices of 50 OSCC cases and 10 normal cases, and the results was treated with statistical lysis.

RESULTSIn 50 OSCC cases, Cyclin D1 mainly manifested in karyon, and a little in cytoplasm. manifesting rates of Cyclin D1 in the samples was 80.0%, which was significantly higher than the manifesting of 20.0% in normal oral mucous membrane (P < 0.01). The manifestation of Cyclin D1 was correlated with rent pathological grades, clinical phases and lymph node metastasis (P < 0.05).

CONCLUSIONThe abnormal tation of Cyclin D1 is closely related with the occurrence and development of OSCC. Therefore, it can subsidiary index for OSCC treatment and prognosis.

Carcinoma, Squamous Cell ; Cyclin D1 ; Female ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; Mouth Mucosa ; Mouth Neoplasms ; Prognosis

OBJECTIVETo analyze the risk factors related to in-hospital bleeding for patients with acute coronary syndrome (ACS).

METHODSClinical and therapeutic data of 3807 patients who were registered with acute coronary syndrome in SINO-GRACE in China from March 2001 to December 2007 were reviewed. A total of 57 patients were grouped to bleeding group and 234 out of the remaining 3750 patients without bleeding were randomly chosen and served as non-bleeding group. Hemorrhage-related factors were screened and compared between the two groups. Unitary logistic regression analysis was performed to detect the possible factors related to hemorrhage. Factors with P < 0.1 were further analyzed by stepwise regression method and multivariate conditional logistic regression analyses.

RESULTS(1) Age, history of coronary artery bypass graft (CABG), previous hemorrhage, renal failure and heart failure as well incidence of acute coronary syndrome were significantly higher in bleeding group than in non-bleeding group (all P ≤ 0.05). Patients were more often treated with clopidogrel and glycoprotein (GP) IIb/IIIa receptor antagonist in bleeding group than in non-bleeding group. (2) Single factor logistic regression analysis showed that age > 70 years, history of previous bleeding, renal failure, heart failure, clopidogrel and GP IIb/IIIa receptor antagonists use, non-ST-segment elevation myocardial infarction, inferior wall, lateral myocardial infarction, CABG were risk factors for bleeding (all P < 0.05). (3) Multivariate logistic regression analysis showed that history of renal failure (OR = 19.77, 95%CI 4.38 - 89.18, P < 0.01) and clopidogrel (OR = 19.77, 95%CI 4.38 - 89.18, P < 0.01) and GPIIb/IIIa receptor antagonist (OR = 343.57, 95%CI 40.39 - 999.99, P < 0.01) use were the independent risk factors for bleeding.

CONCLUSIONOur results show that renal failure history and clopidogrel and GPIIb/IIIa receptor antagonist use are independent risk factors for in-hospital bleeding in patients with acute coronary syndrome.

Acute Coronary Syndrome ; complications ; pathology ; Age of Onset ; Aged ; Female ; Hemorrhage ; etiology ; Hospitalization ; Humans ; Incidence ; Logistic Models ; Male ; Middle Aged ; Platelet Glycoprotein GPIIb-IIIa Complex ; antagonists & inhibitors ; Renal Insufficiency ; Risk Factors ; Ticlopidine ; analogs & derivatives ; therapeutic use

AIM To study the effects of Danggui Buxue Decoction total glycosides combined with Hirudo in the treatment of pulmonary fibrosis in rats and its mechanism.METHODS One hundred and twenty male SD rats were selected,eighteen of which were taken as normal group (normal saline),the remaining 102 rats were used to establish model for pulmonary fibrosis.Seventy-two modeled rats were randomly divided into model group (normal saline),Danggui Buxue Decoction total glycosides group (13 mg/kg),Hirudo group (Hirudo powder,4 g/kg) and combination group (Danggui Buxue Decoction total glycosides,13 mg/kg;Hirudo powder,4 g/kg),eighteen rats in each group with intragastric administration.RESULTS On the 7th,14th and 28th days after modeling,alveolar inflammatory score,pulmonary fibrosis degree score,TGF-β1 and PAI-1 expression levels in lung tissue,IL-4 and IL-17 expression levels in alveolar lavage fluid,hydroxyproline (HYP) content in lung tissue in the model group were significantly higher than those in the normal group (P ＜ 0.05).Alveolar inflammatory score,pulmonary fibrosis degree score,TGF-β1,PAI-1,IL-4,IL-17 expression levels and HYP content in the Danggui Buxue Decoction total glycosides group,Hirudo group and combination group were significantly lower than those in the model group (P ＜ 0.05).In the combination group,alveolar inflammatory score,pulmonary fibrosis degree score,TGF-β1,PAI-1,IL-4,IL-17 expression levels and HYP content were significantly lower than those in the Danggui Buxue Decoction total glycosides group and Hirudo group (P ＜ 0.05).CONCLUSION Danggui Buxue Decoction total glycosides combined with Hirudo have good therapeutic effects on pulmonary fibrosis in rats mainly by reducing TGF-β1,PAI-1 expression levels and HYP coment in lung tissue.

OBJECTIVETo investigate the influence of the lipopolysaccharide receptor CD14-159C/T gene polymorphism on the synthesis and release of high mobility group box-1 protein (HMGB1), and its relation to sepsis in patients with severe burn.

METHODSVenous blood from 35 patients with burn area equal to or larger than 30% TBSA was obtained on post burn day (PBD) 1, 3, 5, 7, 14, 21, and 28 respectively. Eleven volunteers were enrolled as healthy control group (HC).CD14-159C/T gene polymorphism was detected with polymerase chain reaction (PCR)-restriction fragment length polymorphism analysis. Plasma level of HMGB1 was determined with ELISA. Leukocyte HMGB1 mRNA expression was determined with RT-PCR. Data were processed with chi(2) test, analysis of variance, and t test.

RESULTSAmong the C-159T genotype of CD14 gene in the 35 patients, the distribution frequency of the T and the C allele was respectively 57.2% and 42.8%. Seven cases (20.0%) were homozygous for the C allele (CC), 16 cases (45.7%) were heterozygous (TC), and 12 cases (34.3%) were homozygous for the T allele (TT). Allele and genotype frequencies in cases were testified as reaching the Hard-Weinberg equilibrium. The incidence of sepsis was markedly lower in CC homozygous patients than in TC heterozygous and TT homozygous patients. Only one of the 3 septic patients in CC homozygous type died; 4 of 9 septic cases in TC heterozygous type and 4 of 7 septic cases in TT homozygous type died. Plasma levels of HMGB1 of patients were significantly elevated early on PBD 1 as compared with HC group, and higher values were found in TC heterozygous and TT homozygous patients than that in CC homozygous patients on PBD 14, 21, 28 (with F value respectively 3.5671, 4.2035, 3.8529, P < 0.05 or P < 0.01). Higher HMGB1 mRNA expression was found in septic patients as compared with non-sepsis patients on PBD 14 (1.5 +/- 0.5 vs. 1.2 +/- 0.4, t = -2.205, P < 0.05). Plasma level of HMGB1 was also respectively higher in septic patients than in non-sepsis patients on PBD 7, 21 [(44 +/- 29) ng/mL vs. (26 +/- 12) ng/mL, t = -2.355, P < 0.05; (25 +/- 15) ng/mL vs. (10 +/- 6) ng/mL, t = -3.872, P < 0.01)].

CONCLUSIONSCD14C-159T gene polymorphism might markedly influence the synthesis and release of HMGB1, and it is associated with increase in susceptibility of sepsis in patients with severe burn.

Adolescent ; Adult ; Burns ; genetics ; metabolism ; Case-Control Studies ; Disease Susceptibility ; Female ; Gene Frequency ; Genotype ; HMGB1 Protein ; metabolism ; Humans ; Lipopolysaccharide Receptors ; genetics ; Male ; Middle Aged ; Polymorphism, Genetic ; Sepsis ; etiology ; Young Adult

Objective: To prepare monoclonal antibodies (McAb) with cardiac troponin I (cTnI) which was purified from fresh human cardiac muscle within 6 h. Methods: (1) Extraction and purification of human cTnI: cTnI was purified by high salt extraction, saltless precipitation, 65℃ treatment, ammonium sulfate fractionation and DEAE-cellulose chromatography, etc. (2) Preparation of anti human cTnI McAb: The purified cTnI was injected into the spleen of BALB/c mice. The cTnI-primed spleen cells were fused with Sp2/0 myoloma cell. The McAbs anti human cTnI were obtained by screening with indirect ELISA and 3 times clone. (3)The identification of anti cTnI McAb. Results: Five hybridoma cell lines, named 3A7，3A11，3D2，3F10 and 1H9 were developed, which could secret McAb stably. The 5 McAbs all were demonstrated to be IgG2a by double gel diffusion test. The number of hybridoma chromosomes was between 92 to 110 and the chromosomes were mainly telocentric. Five kinds of ascites had no cross-reaction to LDH，CK，CK-MB ，AST and cardiac troponin T(cTnT), and their titers were between 3.2×10-6 to 1.6×10-7. Conclusion: 3D2，3F10 and 3A7,3A11,1H9 react to different epitopes of cTnI.

Objective: To prepare monoclonal antibodies (McAb) with cardiac troponin I (cTnI) which was purified from fresh human cardiac muscle within 6 h. Methods: (1) Extraction and purification of human cTnI: cTnI was purified by high salt extraction, saltless precipitation, 65℃ treatment, ammonium sulfate fractionation and DEAE-cellulose chromatography, etc. (2) Preparation of anti human cTnI McAb: The purified cTnI was injected into the spleen of BALB/c mice. The cTnI-primed spleen cells were fused with Sp2/0 myoloma cell. The McAbs anti human cTnI were obtained by screening with indirect ELISA and 3 times clone. (3)The identification of anti cTnI McAb. Results: Five hybridoma cell lines, named 3A7，3A11，3D2，3F10 and 1H9 were developed, which could secret McAb stably. The 5 McAbs all were demonstrated to be IgG2a by double gel diffusion test. The number of hybridoma chromosomes was between 92 to 110 and the chromosomes were mainly telocentric. Five kinds of ascites had no cross-reaction to LDH，CK，CK-MB ，AST and cardiac troponin T(cTnT), and their titers were between 3.2×10-6 to 1.6×10-7. Conclusion: 3D2，3F10 and 3A7,3A11,1H9 react to different epitopes of cTnI.