Nicotinamide adenine dinucleotide phosphate oxidases (NOXs) are the major enzymatic source of reactive oxygen species (ROS). NOX2 and NOX4 are expressed in the heart but its role in hypoxia-induced atrial natriuretic peptide (ANP) secretion is unclear. This study investigated the effect of NOX on ANP secretion induced by hypoxia in isolated beating rat atria. The results showed that hypoxia significantly upregulated NOX4 but not NOX2 expression, which was completely abolished by endothelin-1 (ET-1) type A and B receptor antagonists BQ123 (0.3 µM) and BQ788 (0.3 µM). ET-1-upregulated NOX4 expression was also blocked by antagonists of secreted phospholipase A2 (sPLA2; varespladib, 5.0 µM) and cytosolic PLA2 (cPLA2; CAY10650, 120.0 nM), and ET-1-induced cPLA2 expression was inhibited by varespladib under normoxia. Moreover, hypoxia-increased ANP secretion was evidently attenuated by the NOX4 antagonist GLX351322 (35.0 µM) and inhibitor of ROS N-Acetyl-D-cysteine (NAC, 15.0 mM), and hypoxia-increased production of ROS was blocked by GLX351322. In addition, hypoxia markedly upregulated Src expression, which was blocked by ET receptors, NOX4, and ROS antagonists. ET-1-increased Src expression was also inhibited by NAC under normoxia. Furthermore, hypoxiaactivated extracellular signal-regulated kinase 1/2 (ERK1/2) and protein kinase B (Akt) were completely abolished by Src inhibitor 1 (1.0 µM), and hypoxia-increased GATA4 was inhibited by the ERK1/2 and Akt antagonists PD98059 (10.0 µM) and LY294002 (10.0 µM), respectively. However, hypoxia-induced ANP secretion was substantially inhibited by Src inhibitor. These results indicate that NOX4/Src modulated by ET-1 regulates ANP secretion by activating ERK1/2 and Akt/GATA4 signaling in isolated beating rat hypoxic atria.

Nicotinamide adenine dinucleotide phosphate oxidases (NOXs) are the major enzymatic source of reactive oxygen species (ROS). NOX2 and NOX4 are expressed in the heart but its role in hypoxia-induced atrial natriuretic peptide (ANP) secretion is unclear. This study investigated the effect of NOX on ANP secretion induced by hypoxia in isolated beating rat atria. The results showed that hypoxia significantly upregulated NOX4 but not NOX2 expression, which was completely abolished by endothelin-1 (ET-1) type A and B receptor antagonists BQ123 (0.3 µM) and BQ788 (0.3 µM). ET-1-upregulated NOX4 expression was also blocked by antagonists of secreted phospholipase A2 (sPLA2; varespladib, 5.0 µM) and cytosolic PLA2 (cPLA2; CAY10650, 120.0 nM), and ET-1-induced cPLA2 expression was inhibited by varespladib under normoxia. Moreover, hypoxia-increased ANP secretion was evidently attenuated by the NOX4 antagonist GLX351322 (35.0 µM) and inhibitor of ROS N-Acetyl-D-cysteine (NAC, 15.0 mM), and hypoxia-increased production of ROS was blocked by GLX351322. In addition, hypoxia markedly upregulated Src expression, which was blocked by ET receptors, NOX4, and ROS antagonists. ET-1-increased Src expression was also inhibited by NAC under normoxia. Furthermore, hypoxiaactivated extracellular signal-regulated kinase 1/2 (ERK1/2) and protein kinase B (Akt) were completely abolished by Src inhibitor 1 (1.0 µM), and hypoxia-increased GATA4 was inhibited by the ERK1/2 and Akt antagonists PD98059 (10.0 µM) and LY294002 (10.0 µM), respectively. However, hypoxia-induced ANP secretion was substantially inhibited by Src inhibitor. These results indicate that NOX4/Src modulated by ET-1 regulates ANP secretion by activating ERK1/2 and Akt/GATA4 signaling in isolated beating rat hypoxic atria.

OBJECTIVESTo evaluate the characteristic morphology of heel spur, and to investigate the relationship of heel spur and plantar heel pain.

METHODSFrom June 2005 to April 2009, 210 cases (254 feet) with heel spur (according to Denis Pain Scale) were divided into cases group 1 (P2, n = 46), 2 (P3, n = 44), 3 (P4, n = 42), 4 (P5, n = 36) and controls group (P1, n = 42). Three-dimensional reconstruction of heel spur was performed in all groups using volume rendering based on multi-slice CT data by Super Image orthopedics edition 1.0. The characteristic morphology of heel spur was observed and the data were measured and analyzed, involving the width of basilar part, the length, the angle between heel spur and planta pedis, and the angle between the longitudinal axis of calcaneus and heel spur.

RESULTSParts of cases groups displayed coarse arcuate edge and undersurface with one or more little heel spurs adhere to heel spur, of which the numbers were greater than controls group, especially in cases group 4. No significant difference of the width of basilar part of heel spur was found among 5 groups (F = 2.32, P > 0.05). However, obvious difference was found in the length, the angle between heel spur and planta pedis, and the angle between the longitudinal axis of calcaneus and heel spur (F = 8.23, 6.82, 5.87, P < 0.05). Compared with the controls group, the angle between heel spur and planta pedis of cases groups had higher degrees, but the difference of the other data presented irregular.

CONCLUSIONSThe characteristic morphology of heel spur varies in patients associated with plantar heel pain. No correlation is found between the severity and the morphological data, including the width of basilar part, the length, the angle between heel spur and planta pedis, and the angle between the longitudinal axis of calcaneus and heel spur.

Aged ; Calcaneus ; diagnostic imaging ; pathology ; Case-Control Studies ; Female ; Heel Spur ; complications ; diagnostic imaging ; pathology ; Humans ; Imaging, Three-Dimensional ; Male ; Middle Aged ; Tomography, X-Ray Computed

OBJECTIVETo study the cell biocompatibility of porous biphasic calcium phosphate nanocomposite in vitro.

METHODSBone marrow mesenchymal cell (BMSCs) obtained from SD rat bone marrow were in vitro induced and proliferated. Afler their osteoblast phenotypes were verified, BMSCs were seeded onto prepared porous biphasic calcium phosphate nanocomposite (Experiment group) and common porous hydroxyapatite (Control group). The cell adhesion was evaluated by scanning electron microscope. Synthesis of alkaline phosphatase enzyme (ALP) and osteocalcin were detected and cell cycle was detected by flow cytometry.

RESULTSBMSCs could fully attach to and extend on the material in experiment and control group, Moreover, experiment group were superior to control group in adhesion, proliferative abilities and osteogenic activity.

CONCLUSIONBMSCs can differentiate to osteoblast phenotype; the porous biphasic calcium phosphate nanocomposite as bone tissue engineering scaffold has good cell biocompatibility.

Alkaline Phosphatase ; Animals ; Bone Marrow Cells ; Bone and Bones ; Cell Adhesion ; Durapatite ; Hydroxyapatites ; Materials Testing ; Nanocomposites ; Osteoblasts ; Osteocalcin ; Rats ; Tissue Engineering ; Tissue Scaffolds

OBJECTIVETo study the feasibility of hand-assisted laparoscopic radical resection of rectal carcinoma and compare the short-term outcomes of HALS versus traditional laparoscopy approach.

METHODSClinical data of 42 cases of rectal carcinoma between January 2010 and March 2011 were enrolled in this study. Nineteen cases underwent HALS total mesorectal excision and 23 cases underwent traditional laparoscopy approach.

RESULTSAll the operations were successfully accomplished without conversions to open surgery. The mean operation time of the HALS group was shorter than that of the traditional laparoscopic group (152 min vs. 168 min, P=0.009). Incision length was significantly longer in the HALS group (5.6 cm vs. 4.5 cm, P=0.000). The median overall costs were lower in HALS group (26 000 RMB vs. 29 000 RMB, P=0.008). The number of lymph nodes in resected specimen, intra-operative blood loss, length of hospital stay, time to passage of flatus were comparable between the two groups.

CONCLUSIONSHand-assisted laparoscopic surgery has the advantages of laparoscopic surgery including minimal invasiveness, safety, and quicker postoperative recovery.

Colectomy ; Hand-Assisted Laparoscopy ; Humans ; Laparoscopy ; Rectal Neoplasms ; surgery ; Treatment Outcome

OBJECTIVETo investigate the relationship between vascular endothelial dysfunction and serum homocysteine (HCY) level in patients with coronary lesions.

METHODSSerum HCY, serum nitric oxide (NO), plasma endothelin-1 (ET-1), and circulation endothelial cell (CEC) were measured in 76 patients who received coronary angiography. Fifty-four patients with a stenosis of 50% or more at least in one coronary atery were as coronary artery disease (CAD) group. Other 22 cases with no recognizable plaque and/or stenosis were as control group. HCY level was detected using an enzyme immunoassay kit. NO concentration was measured using a nitrate reductase kit. Radio-immunoassay was applied to analyse the ET-1 level, and CEC was measured by flow cytometry.

RESULTSThe levels of HCY, ET-1, and CEC in patients with coronary lesions were significantly increased in comparison with control group (P < 0.01), while NO level in CAD group was significantly lower compared with that in control (P < 0.01). Using a multivariate stepwise regression analysis, HCY level had a positive correlation with ET-1 level (r = 0.420, P < 0.05) and CECs number (r = 0.423, P < 0.05); and had a negative correlation with NO/ET-1 (r = -0.403, P < 0.05). But there was no significant correlation between HCY and NO levels.

CONCLUSIONSHCY might lead to endothelial cell injury, which would provide a plausible mechanism for the relationship between hyperhomocysteinemia and development of coronary artery disease. HCY can be considered as a predictor for preliminary or active coronary lesion.

Aged ; Biomarkers ; blood ; Cell Count ; Coronary Artery Disease ; blood ; pathology ; Endothelial Cells ; pathology ; Endothelin-1 ; blood ; Female ; Homocysteine ; blood ; Humans ; Male ; Middle Aged ; Nitric Oxide ; blood

OBJECTIVEInflammatory reaction and injury in immature lungs are associated with activation of nuclear factor-kappa B (NF-kappaB) to trigger proinflammatory cytokine release, but the mechanism thereof is not fully understood. The present study was conducted to understand possible relationship between expression of NF-kappaB and its inhibitor and severity and outcome of neonates with hyaline membrane disease (HMD).

METHODSSerial samples of bronchoalveolar lavage fluid (BALF) were obtained during mechanical ventilation from 31 preterm infants with HMD. These infants were divided into two groups: survivors group [n = 22, birth weight (1500 +/- 320) g and gestational age (31.2 +/- 1.8) weeks] and nonsurvivors group [birth weight (1340 +/- 280) g, gestational age (30.8 +/- 2.1) weeks]. Nineteen preterm infants [birth weight (1470 +/- 280) g, gestational age (30.6 +/- 1.9) weeks] without respiratory disorders were enrolled as control subjects. Alveolar macrophages (AM) were isolated by differential adherence. AM was cultured and treated with lipopolysaccharide (LPS) for 1 hr. Then, nuclear extracts of AM were analyzed by electrophoretic mobility shift assay (EMSA) for NF-kappaB expression. NF-kappaB inhibitor (IkappaB-alpha protein) in cytoplasmic extracts was detected by using Western blotting and IL-1beta and IL-8 in BALF by enzyme-linked immunosorbent assay (ELISA).

RESULTSNF-kappaB complexes were observed by EMSA, they were characterized by competition with cold oligonucleotide and p65-specific antibodies. The addition of an excess of cold oligonucleotide, corresponding to the NF-kappaB binding site, turned off the signal of the band, showing that the band was specific. An excess of an irrelevant oligonucleotide (corresponding to the SP-1) did not show any effect. The addition of an anti-p65 antibody caused the supershift of the two upper bands. After EMSA, the NF-kappaB complexes were quantified by using a ImageQuant software. NF-kappaB expression in AM at 24 hrs was higher in all the patients with HMD as compared with control subjects (survives/control, 34.1 vs 11.4 RDU, P < 0.01; nonsurvivors/control, 55.2 vs 11.4 RDU, P < 0.01). The NF-kappaB expression in AM at 72 hrs was higher than that in control subjects but not for nonsurvivors (survivors/control, 47.8 vs 25.6 RDU, P < 0.01; nonsurvivors/control, 21.8 vs 25.6, P > 0.05). The NF-kappaB expression in AM from nonsurvivors was depressed at 72 hrs as compared to 24 hrs (21.8 vs 55.2, P < 0.01), whereas the NF-kappaB expression in AM from survivors was still higher at 72 hrs than that at 24 hrs (47.8 vs 34.1, t = 4.43, P < 0.01).

CONCLUSIONAltered NF-kappaB activation in AM of BALF of neonates with HMD was observed, and it may be mediated by decreased IkappaB synthesis, increased IkappaB degradation, or both. In HMD nonsurvivors NF-kappaB translocation was hampered upon LPS activation.

Birth Weight ; Blotting, Western ; Bronchoalveolar Lavage Fluid ; cytology ; Cell Culture Techniques ; Cell Nucleus ; drug effects ; metabolism ; Cytoplasm ; drug effects ; metabolism ; Electrophoretic Mobility Shift Assay ; Enzyme-Linked Immunosorbent Assay ; Female ; Gestational Age ; Humans ; Hyaline Membrane Disease ; immunology ; therapy ; I-kappa B Proteins ; immunology ; Infant, Newborn ; Infant, Premature ; immunology ; Interleukin-1beta ; immunology ; Interleukin-8 ; immunology ; Lipopolysaccharides ; pharmacology ; Macrophages, Alveolar ; drug effects ; immunology ; Male ; NF-KappaB Inhibitor alpha ; NF-kappa B ; immunology ; Respiration, Artificial ; Severity of Illness Index ; Time Factors

OBJECTIVETo investigate the diagnostic value of computerized tomographic angiography (CTA) and magnetic resonance angiography (MRA) for intracranial traumatic aneurysms (TAs).

METHODSCTA and MRA of six patients with intracranial TAs verified by digital subtraction angiography (DSA) and surgery were retrospectively analysed. All patients were examined by nonenhanced computerized tomography (CT) and two by CTA. The source data were reconstructed by volume rendering (VR) and multi-planar reconstruction (MPR) from CTA. Four of them had maximum intensity project (MIP) from MRA.

RESULTSOf the six patients, a total of seven TAs were detected by CTA and MRA examinations. Five cases had only one TA and one case had two TAs. The average diameter was 2.3 cm (1.1-3.3 cm). CTA demonstrated two TAs appeared at the cavernous segment of the internal carotid artery (ICA) and the middle cerebral artery (MCA) respectively. MCA TA was definitely and clearly demonstrated on VR images, whereas VR images failed to depict the cavernous ICA TA, which was detected on MPR images. Two TAs were found irregular saccular shape, irregular margin of parent artery and wide neck on CTA. Four MRA examinations demonstrated five TAs, including the cavernous segment ICA TAs (2 cases), the supraclinoid segment ICA TA (1 case), and the cavernous segment associated with opposite side of the petrosal segment ICA TA (1 case). In a cavernous ICA TA, MRA only revealed aneurysm body, whereas aneurysm neck and distal segment of the parent artery were not revealed. In the remaining cases, MRA clearly depicted aneurysm body and parent artery, whereas the neck was not displayed. ICA TAs showed irregular capsule-like high signal intensity on MRA images. Four TAs exhibited irregular distal segment of the parent artery. TAs at the supraclinoid segment or MCA failed to find fracture signs on nonenhanced CT.

CONCLUSIONSBoth CTA and MRA examinations are the effective non-invasive method of imageology for diagnosing intracranial TAs, while CTA is more eligible for diagnosing TAs after nonenhanced CT has demonstrated skull base fractures.

Adult ; Aged ; Brain Injuries ; diagnosis ; Cerebral Angiography ; Female ; Humans ; Intracranial Aneurysm ; diagnosis ; Magnetic Resonance Angiography ; Male ; Middle Aged ; Retrospective Studies ; Tomography, X-Ray Computed

OBJECTIVETo isolate and chondro-inductive culture of human adipose tissue-derived stromal cells and to study their heterotopic chondrogenesis by loading them on alginate gel.

METHODSLiposuction human adipose tissues were minced and digested with collagenase type I. The obtained stromal cells were primarily cultured in BGJb medium for ten days. Secondary harvested cells were cultured in DMEM-F12 medium supplemented with 10%FBS, 6.25 mg/L insulin, 10 mg/L TGF-beta1, 50 mg/L of freshly prepared L-ascorbate for 14 days. After in vitro assay of chondrogenic phenotypes, the cells at density of 10(10)/L were mixed with 1.2% alginate sodium and 102 mmol/L CaCl(2). The cross-linking cell-alginate gel were injected into four BALB/C athymic mice subcutaneously (1 ml for each mouse). Meanwhile, the auto-controls were set by injecting equal dose of simple alginate gel and pure cells in two opposite buttocks of the same mouse subcutaneously. Two mice were sacrificed at fourth and eighth week postoperatively and all samples were removed, fixed, embedded in paraffin and cut into sections of 5 micro m thick. HE staining, Alcian blue and modified Masson's trichrome staining were employed to observe chondrogenesis histologically.

RESULTSAlcian blue and immunocytochemical staining revealed chondroitin sulfate and collagen II in cell matrix after having been chondro-inductive cultured for 14 days. At intervals of fourth and eighth week, heterotopic chondrogenesis is (cartilage formed) within cell-alginate injected sites were found in all mice but negatively in auto-controls. Histologically the hypertrophic chondrocytes were among cartilage matrix in different staining. All alginate gel and solitory cells absorbed within two to three weeks postoperatively in auto-controls.

CONCLUSIONIt seems that stromal cells derived from human adipose tissue presents a potential for chondrogenic differentiation.

Adipose Tissue ; cytology ; Alginates ; pharmacology ; Animals ; Cell Differentiation ; Cells, Cultured ; Chondrocytes ; cytology ; metabolism ; Chondrogenesis ; Female ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Stem Cell Transplantation ; Stromal Cells ; cytology ; metabolism ; transplantation ; Tissue Engineering

OBJECTIVETo investigate the effect of Qingre Quyu Granule (清热祛瘀颗粒, QRQYG) on stabilizing vulnerable plaques in apolipoprotein E (ApoE) deficient mice.

METHODSSeventy-two male ApoE deficient mice were given a high-fat diet from 6 weeks of age. At the 16th week, all the mice were randomized into 3 groups: the QRQYG group, the simvastatin group, and the control group. Sixteen weeks after administration of 0.9 g/kg QRQYG, 3 mg/kg simvastatin or 10 mg/kg sodium chloride per day to the respective groups, the animals were euthanized. The pathological morphologic changes in the vulnerable plaques were evaluated, the matrix metalloprotease-9 (MMP-9) expression was measured by immunohistofluorescence, the soluble intercellular adhesion molecule 1 (ICAM-1) was determined by ELISA, the nuclear factor kappaB (NF-κB) subunit p65 was measured by quantitative RT-PCR, and, finally, thrombospondin-1 (TSP-1) was determined by the immunohistochemical method.

RESULTSThe plaque cross-sectional area in the brachiocephalic artery (23.7%, P<0.01), the lipid core of the plaque (43.1%±3.1%), and the number of buried fibrotic caps of the plaque were significantly decreased in the QRQYG group compared to the control group (both P<0.01); furthermore, the thickness of the fibrotic cap of the plaque increased and the intra-plaque hemorrhage of the plaque decreased. The serum soluble ICAM-1 (27.1±5.1 μg/mL), the protein expression of MMP-9 and TSP-1 and the p65 mRNA expression increased in the QRQYG group in comparison with the control group (P<0.05 or P<0.01).

CONCLUSIONQRQYG could stabilize the vulnerable plaque through inhibition of the inflammatory response.

Animals ; Apolipoproteins E ; genetics ; Atherosclerosis ; pathology ; Brachiocephalic Trunk ; drug effects ; enzymology ; metabolism ; pathology ; Drugs, Chinese Herbal ; pharmacology ; Enzyme-Linked Immunosorbent Assay ; Intercellular Adhesion Molecule-1 ; metabolism ; Male ; Matrix Metalloproteinase 9 ; metabolism ; Mice ; Mice, Knockout ; NF-kappa B ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Simvastatin ; pharmacology ; Sodium Chloride ; pharmacology ; Thrombospondin 1 ; metabolism