S-nitrosylation of AMPKγ impairs coronary collateral circulation and disrupts VSMC reprogramming.

Collateral circulation is essential for blood resupply to the ischemic heart, which is dictated by the contractile phenotypic restoration of vascular smooth muscle cells (VSMC). Here we investigate whether S-nitrosylation of AMP-activated protein kinase (AMPK), a key regulator of the VSMC phenotype, impairs collateral circulation. In rats with collateral growth and development, nitroglycerin decreases coronary collateral blood flow (CCBF), inhibits vascular contractile phenotypic restoration, and increases myocardial infarct size, accompanied by reduced AMPK activity in the collateral zone. Nitric oxide (NO) S-nitrosylates human recombinant AMPKγ1 at cysteine 131 and decreases AMP sensitivity of AMPK. In VSMCs, exogenous expression of S-nitrosylation-resistant AMPKγ1 or deficient NO synthase (iNOS) prevents the disruption of VSMC reprogramming. Finally, hyperhomocysteinemia or hyperglycemia increases AMPKγ1 S-nitrosylation, prevents vascular contractile phenotypic restoration, reduces CCBF, and increases the infarct size of the heart in Apoe-/- mice, all of which is rescued in Apoe-/-/iNOSsm-/- mice or Apoe-/- mice with enforced expression of the AMPKγ1-C130A mutant following RI/MI. We conclude that nitrosative stress disrupts coronary collateral circulation during hyperhomocysteinemia or hyperglycemia through AMPK S-nitrosylation.

S-Nitrosylation of Akt by organic nitrate delays revascularization and the recovery of cardiac function in mice following myocardial infarction.

The effects of long-term nitrate therapy are compromised due to protein S-Nitrosylation, which is mediated by nitric oxide (NO). This study is to determine the role of Akt S-Nitrosylation in the recovery of heart functions after ischaemia. In recombinant Akt protein and in HEK293 cells, NO donor decreased Akt activity and induced Akt S-Nitrosylation, but was abolished if Akt protein was mutated by replacing cysteine 296/344 with alanine (Akt-C296/344A). In endothelial cells, NO induced Akt S-Nitrosylation, reduced Akt activity and damaged multiple cellular functions including proliferation, migration and tube formation. These alterations were ablated if cells expressed Akt-C296/344A mutant. In Apoe-/- mice, nitroglycerine infusion increased both Akt S-Nitrosylation and infarct size, reduced Akt activity and capillary density, and delayed the recovery of cardiac function in ischaemic hearts, compared with mice infused with vehicle. Importantly, these in vivo effects of nitroglycerine in Apoe-/- mice were remarkably prevented by adenovirus-mediated enforced expression of Akt-C296/344A mutant. In conclusion, long-term usage of organic nitrate may inactivate Akt to delay ischaemia-induced revascularization and the recovery of cardiac function through NO-mediated S-Nitrosylation.

Molecular Evolution and Emergence of H5N6 Avian Influenza Virus in Central China.

H5N6 avian influenza virus (AIV) has posed a potential threat to public health since its emergence in China in 2013. To understand the evolution and emergence of H5N6 AIV in the avian population, we performed molecular surveillance of live poultry markets (LPMs) in Wugang Prefecture, Hunan Province, in central China, during 2014 and 2015. Wugang Prefecture is located on the Eastern Asian-Australian migratory bird flyway, and a human death due to an H5N6 virus was reported in the prefecture on 21 November 2016. In total, we sampled and sequenced the complete genomes of 175 H5N6 AIVs. Notably, our analysis revealed that H5N6 AIVs contain at least six genotypes arising from segment reassortment, including a rare variant that possesses an HA gene derived from H5N1 clade 2.3.2 and a novel NP gene that has its origins with H7N3 viruses. In addition, phylogenetic analysis revealed that genetically similar H5N6 AIVs tend to cluster according to their geographic regions of origin. These results help to reveal the evolutionary behavior of influenza viruses prior to their emergence in humans.IMPORTANCE The newly emerged H5N6 influenza A virus has caused more than 10 human deaths in China since 2013. In November 2016, a human death due to an H5N6 virus, in Wugang Prefecture, Hunan Province, was confirmed by the WHO. To better understand the evolution and emergence of H5N6 viruses, we surveyed live poultry markets (LPMs) in Wugang Prefecture before the reported human death, with a focus on revealing the diversity and genomic origins of H5N6 in birds during 2014 and 2015. In general, H5N6 viruses in this region were most closely related to H5N1 clade 2.3.4.4, with the exception of one virus with an HA gene derived from clade 2.3.2 such that it represents a novel reassortant. Clearly, the ongoing surveillance of LPMs is central to monitoring the emergence of pathogenic influenza viruses.

Genetic properties and pathogenicity of a novel reassortant H10N5 influenza virus from wild birds.

In this study, we analyzed the genome of a H10N5 influenza virus from wild birds. This virus was identified as a novel reassortant virus with internal genes from multiple subtypes and of distinct origins. After sequential passage in mice, mouse-adapted viruses bearing mutations PB2-E627K and HA-G218E were generated. These viruses caused dramatic body weight loss and death, and also replicated in mouse brain, suggesting that the pathogenicity of low pathogenic H10N5 in chickens can be enhanced after passage in mammals. Our data imply that H10N5 viruses might be a potential risk to human health therefore it is important to undertake continued surveillance and biosecurity evaluation of these viruses.

EFFECT OF COLLA CORNUS CERVI COMBINED WITH LV-MEDIATED BMP7 TRANSFECTED BMSCs ON ANFH IN RATS.

In the present study, we investigated the combined effect of Colla Comus Cervi (CCC) and BMP7-overexpressing bone marrow-derived mesenchymal stem cells (BMSCs) on osteogenic induction and the treatment of avascular necrosis of the femoral head (ANFH). BMSCs were isolated from rats. BMP7-overexpressing BMSCs were generated by lentiviral-mediated gene transduction. Cell proliferation, alkaline phosphatase (ALP) activity, osteogenesis related gene expression, osteocalcin levels, and calcified nodules were quantified and compared between four groups: untreated controls, BMSCs cultured with CCC complex medium, BMP7-overexpressing BMSCs, and BMP7-overexpressing BMSCs cultured with CCC complex medium (CCC+BMP7). CCC+BMP7 BMSCs showed higher proliferation rate. ALP activity and osteaocalcin content were significantly increased in CCC+BMP7 BMSCs. The osteogenesis related genes, COLI, and integrin-α2, -α5, and -ß1, were expressed significantly higher in CCC+BMP7 BMSCs. The number of calcified nodules in the CCC+BMP7 group was significantly higher than that in other groups. For in vivo assays, ANFH was induced in rats, and BMSCs were injected into the femoral head of the lower left extremity. In rats with induced ANFH, general observation scores of the CCC+BMP7 injected group were significantly higher than the model group. X-ray and microscopic observations revealed that ANFH was significantly improved and femoral head cells gradually recovered in rats treated with CCC+BMP7 BMSCs. Our results suggest that CCC+BMP7 significantly promote the proliferation and osteogenic differentiation of BMSCs in vitm. CCC+BMP7 BMSCs promote the ability of repairing ANFH in rats, providing a new therapeutic paradigm for the treatment of ANFH.

Inferior ST segment elevation myocardial infarction combined with hyperextension cervical spine injury: A rare case report.

RATIONALE: Inferior ST-segment elevation myocardial infarction (STEMI) is usually caused by acute occlusion of the right coronary artery or left circumflex coronary artery (LCX). Inferior STEMI can cause a high-grade atrioventricular block, sinus bradycardia, and hypotension, leading to hemodynamic collapse, syncope, and disturbance of consciousness. PATIENT CONCERNS: A case of a sudden disturbance of consciousness after chest tightness as the initial symptom, followed by incomplete paralysis and paresthesia of the extremities due to the collision of the face with the ground. DIAGNOSES: Coronary angiography showed about 99% of stenosis in the LCX. Cervical spine magnetic resonance imaging showed C2/3, C3/4, C4/5, and C5/6 intervertebral disc herniation with secondary spinal stenosis, spinal cord compression, and edema. The patient was diagnosed with inferior STEMI combined with hyperextension cervical spine injury. INTERVENTIONS AND OUTCOMES: Bivalirudin was used for anticoagulation, the LCX lesion was pre-expanded with a balloon and the thrombus was removed, and anti-platelet aggregation therapy was given postoperatively. After rehabilitation therapy, hyperextension cervical spine injury improved. There was no recurrence of syncope and precordial pain during the 6-month follow-up. LESSONS: Hyperextension cervical spine injury has unique hemodynamic features that mimic those associated with inferior STEMI, so a detailed medical history inquiry and physical examination should be carried out to avoid missed diagnoses.

Serum­derived exosomal hsa­let­7b­5p as a biomarker for predicting the severity of coronary stenosis in patients with coronary heart disease and hyperglycemia.

Exosomal microRNAs (miRNAs/miRs) are potential biomarkers for the diagnosis and treatment of cardiovascular disease, and hyperglycemia serves an important role in the development of atherosclerosis. The present study aimed to investigate the expression profile of serum­derived exosomal miRNAs in coronary heart disease (CHD) with hyperglycemia, and to identify effective biomarkers for predicting coronary artery lesions. Serum samples were collected from eight patients with CHD and hyperglycemia and eight patients with CHD and normoglycemia, exosomes were isolated and differentially expressed miRNAs (DEMIs) were filtered using a human miRNA microarray. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed using standard enrichment computational methods for the target genes of DEMIs. Receiver operating characteristic (ROC) curve analysis was applied to evaluate the values of the selected DEMIs in predicting the severity of coronary stenosis. A total of 10 DEMIs, including four upregulated miRNAs (hsa­let­7b­5p, hsa­miR­4313, hsa­miR­4665­3p and hsa­miR­940) and six downregulated miRNAs (hsa­miR­4459, hsa­miR­4687­3p, hsa­miR­6087, hsa­miR­6089, hsa­miR­6740­5p and hsa­miR­6800­5p), were screened in patients with CHD and hyperglycemia. GO analysis showed that the 'cellular process', 'single­organism process' and 'biological regulation' were significantly enriched. KEGG pathway analysis revealed that the 'mTOR signaling pathway', 'FoxO signaling pathway' and 'neurotrophin signaling pathway' were significantly enriched. Among these DEMIs, only hsa­let­7b­5p expression was positively correlated with both hemoglobin A1C levels and Synergy between Percutaneous Coronary Intervention with Taxus and Cardiac Surgery score. ROC curves showed that hsa­let­7b­5p could serve as an effective biomarker for differentiating the severity of coronary stenosis. In conclusion, the present study demonstrated that serum­derived exosomal hsa­let­7b­5p is upregulated in patients with CHD and hyperglycemia, and may serve as a noninvasive biomarker for the severity of coronary stenosis.

Vitamin E ameliorates ox-LDL-induced foam cells formation through modulating the activities of oxidative stress-induced NF-κB pathway.

The role of antioxidant supplementation with vitamin E in the prevention of atherosclerosis has been a topic of considerable recent interest. The relevance of vitamin E for macrophage-derived foam cell formation, a hallmark of atherosclerosis, however, has not been unequivocally resolved. Here, we investigated the effect of oxidized LDL (ox-LDL) and vitamin E on lipid accumulation and total cholesterol content in U937 macrophages, reactive oxygen species generation and expression of nuclear factor-κB (NF-κB) signaling pathway. The results showed that the mRNA expression and protein levels of P-selectin were evident in U937 macrophages treated with ox-LDL and vitamin E, which indicating that expression of P-selectin is important in macrophage-derived foam cell formation. Moreover, P-selectin changes in ox-LDL-induced foam cell formation can be mediated by vitamin E through activities of nuclear NF-κB activated by serine phosphorylation of NF-κB inhibitor α, suggesting that activation of NF-κB pathway by macrophages may occur. Taken together, these data suggested that vitamin E can prevent ox-LDL-induced foam cell macrophages formation through modulating the activities of oxidative stress-induced NF-κB pathway.

The effects of physical activity on physiological markers in breast cancer survivors: A meta-analysis.

BACKGROUND: To systematically evaluate the effects of physical activity on physiological markers in breast cancer survivors. METHODS: A systematic search of the PubMed, Wed of Science, Medline, CNKI and Wanfang Database was performed to identify eligible randomized controlled trials to explore physical activity on physiological markers in breast cancer survivors. STATA version 13.0 (Stata Corp LP, College Station, TX) was used for all statistical analyses. RESULTS: A total of 11 articles with 941 cases were eligible in this meta-analysis. The results of the meta-analysis showed that physical activity could decrease the levels of insulin (SMD = -1.90, 95%CI: -3.2 to -0.60; I = 92.3%, P < .001), insulin-like growth factor 1 (IGF-I) (WMD = -4.67, 95%CI: -23.14 to 13.79; I = 96.2%, P < .001), insulin-like growth factor binding protein-3 (IGFBP-3) (WMD = -20.09, 95%CI: -47.15 to 6.97; I = 93.3%, P < .001). However, compared with the control group, there was not the significant change of insulin-like growth factor 2 (IGF-II), insulin-like growth factor binding protein-1 (IGFBP-1), leptin, adiponectin, glucose, C-reactive protein (CRP), Interleukin-6 (IL-6), Interleukin-10 (IL-10), and tumor necrosis factor alpha (TNF-ɑ) levels after the intervention. CONCLUSIONS: Physical activity could improve the insulin function that might be associated with decreasing the levels of IGF-I, IGFBP-3 and insulin in breast cancer survivors.

Tissue-engineered skin containing mesenchymal stem cells improves burn wounds.

There is increasing evidence showing that adult stem cells are useful for tissue regeneration. Bone marrow mesenchymal stem cells (MSCs) are self-renewing and are potent in differentiating into multiple cells and tissues. To investigate the practicability of repairing burn wounds with tissue-engineered (TE) skin combined with bone MSCs, we established a burn wound model in the porcine skin. With a controlling temperature and time of the burning device to obtain different degrees of burn wounds, a deep dermal partial thickness burn was introduced to the porcine skin using a heated-brass contact injury at 100 degrees C for 20 s. Collagen-GAG scaffolds were utilized as the matrix; MSCs separated from pigs were seeded on them to form the skin equivalent. When grafted to the burn wounds, the TE skin containing MSCs showed better healing and keratinization, less wound contraction, and more vascularization. Grafts proliferated well and contributed to the neo-tissues. These data suggest that TE skin containing MSCs in a burn defect can accelerate wound healing and receive satisfactory effects.

[Imbalance of T helper 1 cells/T helper 2 cells accelerated T-cell-mediated endothelium injury in patients with acute coronary syndromes].

OBJECTIVE: To observe the T helper 1 and T helper 2 (Th1/Th2) balance and possible association to vascular endothelial cells injury in patients with acute coronary syndromes (ACS). METHODS: Forty patients with ACS and 18 patients with stable angina pectoris (SAP) were included in this study. The concentrations of T helper 1/T helper 2 subsets related cytokines in plasma were evaluated by ELISA Kits. Cytotoxic activity of peripheral blood mononuclear cells (PBMCs) or PBMCs depleted CD(+) T cells against human umbilical vein endothelial cells (HUVECs) were evaluated by Cr51 cytotoxicity assay. RESULTS: Concentrations of T helper 1 related cytokines IFN-gamma and IL-2 were significantly higher [IFN-gamma: (131.2 +/- 42.2) ng/L vs. (47.6 +/- 20.2) ng/L; IL-2: (83.7 +/- 21.3) ng/L vs. (46.2 +/- 16.7) ng/L, all P < 0.05] while T helper 2 related cytokine IL-10 concentration was significantly lower [(16.7 +/- 4.3) ng/L vs. (27.5 +/- 5.5) ng/L, P < 0.05] in patients with ACS compared to those in SAP patients. Cytotoxic activity of PBMCs against HUVECs in patients with ACS was also significantly higher than that in patients with SAP (28.84% +/- 4.20% vs. 20.28% +/- 2.71%, P < 0.05). CONCLUSIONS: In patients with ACS, Th1 related cytokines were significantly upregulated while Th2 related cytokines were significantly downregulated. This imbalance of Th1/Th2 accelerated PBMCs mediated endothelium injury in patients with ACS.

Alpha-linolenic acid attenuates high glucose-induced apoptosis in cultured human umbilical vein endothelial cells via PI3K/Akt/eNOS pathway.

OBJECTIVE: High glucose-induced apoptosis in vascular endothelial cells contributes to the acceleration of atherosclerosis associated with diabetes. We hypothesized that alpha-linolenic acid (ALA) might attenuate high glucose-induced apoptosis in cultured human umbilical vein endothelial cells (HUVECs). METHODS: HUVECs were cultured at 5.5 and 33 mmol/L for 72 h. ALA with different concentrations was added with defatted bovine serum albumin as a carrier for 18 h before incubation with high glucose. RESULTS: Exposure of HUVECs to high glucose media for 72 h significantly increased the number of apoptotic cells compared with normal glucose control, as evaluated by flow cytometry and terminal deoxyuridine triphosphate nick end labeling assay. Pretreatment with low concentrations of ALA (10, 50, and 100 micromol/L) significantly attenuated high glucose-induced apoptosis of HUVECs, but increasing ALA to 200 micromol/L exerted the opposite effect. Furthermore, high glucose reduced phosphorylation of Akt and endothelial nitric oxide synthase (eNOS) with subsequent nitric oxide production, whereas ALA treatment attenuated the reduction caused by high glucose. Pretreatment with phosphatidylinositol 3' -kinase kinase inhibitor LY294002 and eNOS inhibitor N(G)-nitro-arginine methyl ester eliminated ALA' antiapoptotic effect. CONCLUSION: ALA exerts an antiapoptotic effect by the phosphatidylinositol 3'-kinase/Akt/eNOS pathway in HUVECs exposed to high glucose and thus may represent a candidate therapeutic agent for diabetic cardiovascular complications.

The significance of ENAH in carcinogenesis and prognosis in gastric cancer.

The ENAH gene, which encodes a member of the enabled/vasodilator-stimulated phosphoprotein (Ena/VASP) family of proteins, is involved in the assembly of actin filaments required for cell adhesion and motility. Recent studies show overexpressed ENAH in several cancer types, and ENAH correlates with tumor invasiveness. This study aimed to investigate the expression and function of ENAH in primary gastric adenocarcinoma, and its prognostic significance. We found significantly increased mRNA (P = 0.0283) and protein (P = 0.0301) expression of ENAH in gastric cancer tissues. ENAH expression markedly associated with tumor size (P < 0.001), T stage (P < 0.001), N stage (P = 0.001), TNM stage (P < 0.001) and prognosis (P < 0.001). Cox regression analyses revealed ENAH expression as an independent predictor of overall survival (P = 0.019). We also analyzed data of 155 gastric cancer cases from The Cancer Genome Atlas (TCGA) and found that ENAH expression significantly correlated with age (P = 0.003), T stage (P = 0.023) and prognosis (P = 0.05). Furthermore, the function of ENAH in cell proliferation, colony formation, cell migration and invasion of gastric cancer cells was analyzed in vitro. Knockdown of ENAH expression suppressed cell proliferation, colony formation, cell migration and invasion in MKN45 cells. Conversely, overexpression of ENAH promoted cell proliferation, cell migration and invasion in MGC803 cells. Our research suggests that ENAH might play promoting functions in carcinogenesis and progression of gastric cancer, and may serve as a valuable prognostic marker for primary gastric adenocarcinoma patients.

RELMα can cause contraction of rat aortic smooth muscle cells.

OBJECTIVE: This study aims to observe the contraction role of RELMα in rat aortic smooth muscle cells and explore its mechanism. METHODS: Rat aortas smooth muscle cells were cultivated using tissue explants method. They were divided into 5 groups. A: Control group; B: 1×10(-7) mol/L ANGII group; C: 1×10(-8) mol/L RELMα group; D: 2×10(-8) mol/L RELMα group; E: 4×10(-8) mol/L RELMα group. The thoracic aortic tension signal of rats was recorded by Powerlab system. The expression levels of CaM and MLCK were detected by western blotting and RT-PCR methods. RESULTS: Tension changes of rat thoracic aorta vascular ring in group C, D and E (72±2.98%, 76.65±2.73%, 85.07±3.06% respectively) were higher than that of group A and B (6.35±0.75%, 61.47±4.47%) with dose-dependent (P<0.01). The expression levels of CaM and MLCK in group C were higher than that of group A and B while were lower than that of group D and E (P<0.05). The expression levels of CaM and MLCK in group E were the highest among the groups (P<0.05). CONCLUSIONS: RELMα can cause contraction of rat aortic smooth muscle cells, its mechanism may be via Ca(2+)-CaM-MLCK pathway.

Value of segmental coronary calcium score on diagnosis and interventional treatment of coronary lesions by 320-slice DVCT.

UNLABELLED: The global coronary calcium score has been widely used in the evaluation of coronary plaque burden and cardiovascular disease events. In this study, we investigated the value of segmental coronary calcium score (SCCS) on the diagnosis and interventional treatment. We studied 87 patients with coronary angiography (CAG) and coronary CT angiography (CTA) by 320-slice dynamic volume CT (DVCT). SCCS was determined for each segmental separately. All lesions which SCCS was greater than 0 were enrolled, and were divided into three groups, mild calcification group (SCCS were less than 80), Moderate calcification group (SCCS were more than 80 and less than 200) and Severe calcification group (SCCS were more than 200). From above three groups, lesions received the intervention treatment were elected as subgroup. The position of lesions, plaque morphology, calcification proportion and interventional treatment data were analyzed. Severe calcification group were more frequent in the proximal lesions, stenosis with lesser extent, nubbly and nodular types of plaque, and the inconsistency with CAG was higher than the other two groups (P < 0.05). In the subgroup, more pre-dilatation and post-dilatation balloon were used in severe calcification group, with higher expansion pressure of balloon and stent (P < 0.05), but the diameter of stents was no difference between the three groups. CONCLUSION: SCCS is better than GCCS in the evaluation of coronary calcification, and play an important role in the judgment of stenosis by coronary CT and in the choice of interventional therapeutic devices.

A cross-sectional survey of dental caries, oral hygiene, and Helicobacter pylori infection in adults.

We explored the epidemiological risk factors for dental caries to help explain differences in the prevalence of adult dental caries. We examined 841 people for the presence of Helicobacter pylori in their dental plaque and for dental caries. Of the 841 subjects, 574 (68.25%) were infected with H pylori, and 516 (61.36%) were diagnosed with dental caries. Among the 574 subjects with H pylori, the prevalence of dental caries was 73.52% (422/574), while the prevalence among the 267 cases without H pylori was 35.21% (94/267). A correlation existed between the presence of H pylori and the occurrence of dental caries (χ(2) = 112.8, P < .01, odds ratio = 5.110, 95% confidence interval = 3.740-6.982). The 574 persons with H pylori had a higher mean dental plaque index than those without. In conclusion, H pylori infection in the oral cavity is associated with dental caries and poor dental hygiene.

The opposite-direction modulation of CD4+CD25+ Tregs and T helper 1 cells in acute coronary syndromes.

Different subsets of T lymphocytes have different functions in atherosclerosis advancement. T helper 1 cells and T regulatory 1 cells have been demonstrated to play opposite roles in rupture of atherosclerotic lesion. However, the role of novel subset of T regulatory cells, known as CD4+CD25+Foxp3+ T cells, remains largely unknown in coronary artery disease (CAD). In this study, we investigated the peripheral CD4+CD25+Foxp3+ T cells of patients with CAD and controls. The patients submitted were divided into three groups: stable angina pectoris (SA) group, unstable angina pectoris (UA) group and acute myocardial infarction (AMI) group. We analyzed the frequencies of peripheral CD4+CD25+Foxp3+ T cells and T helper 1/T helper 2 cells, expression of Foxp3 in CD4+CD25+ T subsets and cytokines pattern in patients and controls. We found that the reduction of CD4+CD25+Foxp3+ T lymphocytes was consistent with the expansion of Th1 cells in patients with unstable CAD. The reversed development between CD4+CD25+ Tregs and Th1 cells might contribute to plaque destabilization.