Objective To observe the preventive and therapeutic effects of L-carnitine on the metabolic disorder and cardiac remodeling in alcoholic cardiomyopathy. Methods Experimental animals were divided into three groups: alcohol-fed group(A), an alcohol/L-carnitine fed group(B) and a control group(C). Free fatty acid(FFA) and earnitine were detected in the blood serum at different time. mRNA and protein expressions of peroxisome proliferator-activated receptors (PPARα and PPARγ), retinoic acid receptor retinoid X receptor alpha (RXRα), earnitine palmitoyl transferase isoform and medium-chain acyl-CoA dehydrogenase (MCAD) were observed with RT-PCR and Western blotting methods. Results (1) When group A and B were compared with group C, FFA was increased and carnitine was decreased;mRNA and protein expressions of PPARα, RXRα, CPT-Ⅰ and MCAD were decreased with the development of alcoholic cardiomyopathy (ACM), being more significantly in group A than group B (P < 0.05). (2) mRNA and protein expressions of PPARγ had no statistical significance between these three groups at the end of 2 and 4 months(P>0.05), but after 6 months, they were increased in group B and decreased in group A (A vs. C,P<0.01;B vs. C,P<0.05). Conclusion Metabolic disorder and cardiac remodeling occur in the development process of ACM; they are partly prevented by L-carnitine through downregulating mRNA and protein expressions of PPARct, RXRα, CPT-Ⅰ, MCAD and PPARγ.

OBJECTIVETo establish a localization ultrathin section method through which target cytopathic cells could be sectioned in situ.

METHODSLab-Tek Chamber slide system (177402) was selected as resin embedding mould. Cells infected with Human adenovirus type 5 (Ad5) or A/HN/SWL3/ 2009 (H1N1) influenza virus were embedded in situ as models. Target cytopathic cells were exposed by trimming, sectioned and observed under transmission electron microscope (TEM).

RESULTSTarget cells could be sectioned in situ and virus particles could be found easily on sections.

CONCLUSIONA localization ultrathin sectioning method was established and this technique could be applied in virus detection in cytopathic cells to improve TEM detection efficiency.

Adenovirus Infections, Human ; pathology ; virology ; Adenoviruses, Human ; physiology ; ultrastructure ; Cell Line ; Humans ; Influenza A Virus, H1N1 Subtype ; physiology ; ultrastructure ; Influenza, Human ; pathology ; virology ; Microscopy, Electron, Transmission ; Microtomy ; methods

AIM:To investigate the effect of Astragalus polysaccharides ( APS) on chronic heart failure and its mechanism.METHODS:Male SD rats (n=32) were randomly divided into control group , sham group, model group and APS group (8 rats in each group).The left coronary artery ligation in the rats was conducted to establish myocardial infarc -tion heart failure model.After modeling, the rats in APS group were given APS (3 g· kg-1· d-1) by intragastric adminis-tration for 6 weeks.Left ventricular diastolic diameter (LVD), left ventricular systolic diameter (LVS), left ventricular ejection fraction ( LVEF) and fractional shortening ( FS) were detected by echocardiography .HE staining was used to ob-serve the pathological changes .The concentrations of free fatty acid ( FFA) in the serum and myocardium were observed by the method of acetyl coenzyme A synthetase and acetyl coenzyme A oxidase ( ACS-ACOD ) .The protein levels of total AMP-activated protein kinase (AMPK), phosphorylated AMP-activated protein kinase (p-AMPK), fatty acid translocase (FAT/CD36) and carnitine palmitoyltransferase I (CPT-1) were measured by Western blotting.RESULTS: No signifi-cant difference in each index between sham group and control group was observed .Compared with control group , LVEF and FS in model group was significantly decreased , while LVD and LVS was significantly increased ( P<0.05 ) .The LVEF and FS in APS group were significantly improved compared with model group ( P<0.05 ) , and there was no significant difference between APS group and control group .LVD and LVS in APS group were obviously improved compared with mo-del group (P<0.05), and the difference was significant compared with control group (P<0.05).Compared with control group, focal myocardial necrosis increased , and residual myocardial cells reduced in model group , while those was much better in APS group as compared with model group (P<0.05).The FFA concentrations in the serum and myocardium in model group increased significantly compared with control group ( P<0.05 ) , while those decreased significantly in APS group as compared with model group (P<0.05).The protein levels of p-AMPK, CPT-1, and cell membrane FAT/CD36 in model group decreased significantly compared with control group (P<0.05), and those in APS group increased obvious-ly compared with control group (P<0.05).CONCLUSION:APS improves chronic heart failure by activating the AMPK pathway and promoting myocardial ingestion and utiliation of FFA .

Objective To explore the effects of estrogen on stress-induced senescence of vascular smooth muscle cells (VSMCs) and the underlying mechanisms. Methods The VSMCs of passage 2-3 cultured from female SD rats were induced into senescence by exposing to 150μmol/L H2O2 in the presence or absence of different concentrations(10-10mol/L-10-8mol/L) of 17β-estradiol (E2). The expressions or activities of senescence associated marker DcR2, senescence-associated beta-galactosidase (SA-β-Gal), oncogene Ras and p21WAF1 were detected by flow cytometry, cytochemical staining, pull-down assay or Western blotting analysis. Results Flow cytometry analysis showed that in the physiological concentrations, E2 significantly inhibited the H2O2-promoted high-level expression of DcR2 of VSMCs in a dose-dependent manner, with a highest inhibitive rate at 14.48%±0.6%(E2=10-8 mol/L;P<0.05, n =3);this inhibitive effect could be blocked by a E2 receptors inhibitor ICI 182,780. Cytochemistry staining showed that the rate of SA-β-Gal positive VSMCs induced by H2O2 decreased in presence of 10-8mol/L E2 (20.5%±1.4% vs 9.6%±0.9%;P<0.05, n =9). Pull-down assay and Western blotting analysis revealed that administration of 10-8mol/L E2 obviously reduced the H2O2-induced activity of Ras (0.60±0.06 vs 0.26±0.04;P<0.05, n =3) and expression of p21WAF1 (0.46±0.04 vs 0.33±0.02;P<0.05, n =3). Conclusion E2 exerts, an inhibitive effects on stress-induced senescence of VSMCs by suppressing the activity of Ras and expression of p21WAF1. This finding suggests a novel mechanism for the hormone's anti-atheroschlerotic effects.

[ ABSTRACT] AIM:To study the protective effect of Astragalus polysaccharides ( APS) on free fatty acid-induced injury in human umbilical vein endothelial cells (HUVECs).METHODS: Cultured HUVECs were divided into control group, APS group [ APS (200 mg/L) treated for 24 h], free fatty acid group [free fatty acid (0.25 mmol/L) treated for 24 h], free fatty acid plus APS group [free fatty acid (0.25 mmol/L) and APS (200 mg/L) treated for 24 h], and com-pound C group [ free fatty acid (0.25 mmol/L) and APS (200 mg/L) and AMPK inhibitor compound C (10 μmol/L) treated for 24 h] .The cell viability was detected by MTT assay.Nitric oxide ( NO) content in the medium was determined by nitrate reductase assay.The protein levels of total adenosine monophosphate-activated protein kinase (AMPK), phos-phorylated adenosine monophosphate-activated protein kinase (p-AMPK), endothelial nitric oxide synthase (eNOS) and phosphorylated endothelial nitric oxide synthase ( p-eNOS) were measured by Western blot.RESULTS: No significant difference of all indexes between APS group and control group was observed.The cell viability in free fatty acid group de-creased significantly compared with control group.The cell viability in free fatty acid plus APS group was significantly im-proved as compared with free fatty acid group.The cell viability in compound C group was almost the same as that in free fatty acid group.The No content and protein levels of p-AMPK and p-eNOS in free fatty acid group decreased obviously as compared with control group, while the NO content and protein levels of p-AMPK and p-eNOS in free fatty acid plus APS group increased obviously compared with free fatty acid group.No significant difference of the p-AMPK and p-eNOS protein levels between free fatty acid plus APS group and free fatty acid group was observed.No significant difference of the AMPK and eNOS protein levels in all groups was found.CONCLUSION:APS attenuates the free fatty acid-induced injury, and its mechanism is related to the AMPK-eNOS signal pathway.

Ebola virus (EBOV) and Marburg virus (MARV) belong to the family Filoviridae. Filoviruses cause severe filovirus hemorrhagic fever (FHF) in humans, with high case fatality rates, and represent potential agents for bioterrorism and biological weapons. It is necessary to keep surveillance of filoviruses, even though there is no report of their isolation and patients in China so far. To characterize MARV morphology, the Lake Victoria marburgvirus--Leiden was stained negatively and observed under a transmission electron microscope which is one of important detection methods for filoviruses in emergencies and bioterrorism. MARV showed pleomorphism, with filamentous, rod-shaped, cobra-like, spherical, and branch-shaped particles of uniform diameter but different lengths. Pleomorphism of negatively stained MARV is summarized in this article, so as to provide useful information for possible electron microscopic identification of filoviruses in China.
Animals ; Humans ; Marburg Virus Disease ; virology ; Marburgvirus ; growth & development ; ultrastructure ; Microscopy, Electron, Transmission ; Virion ; growth & development ; ultrastructure

Acute Disease ; Esophageal Stenosis ; chemically induced ; Herbicides ; toxicity ; Humans ; Paraquat ; toxicity ; Poisoning

Objective:To investigate the application of Xingnaojing injection ( XNJI) in the patients of neurosurgery department to provide reference for the rational drug use. Methods:The retrospective investigation was applied to survey 200 hospitalization records with XNJI from a certain hospital between January 2014 and June 2015. The usage, dosage, medication purpose, course of treatment, compatibility and adverse effects of XNJI were analyzed. Results:The unlabeled use of XNJI was found in neurosurgery department, and 34. 50% of the surveyed patients were found that their diagnoses didn’t conform to the indications in the medicine specification of XNJI. Moreover, 92. 50% of the inpatients were treated with overdose and almost 90. 00% were treated with single drug dose of 40 ml. It was also found that XNJI was often combined with potassium chloride or with potassium chloride plus insulin in clinical use. Conclu-sion:There is some irrational use of XNJI in clinics, thus the use and management of traditional Chinese medicine injections should be strengthened and regulated.

Objective To investigate the effect of arginase (Arg) inhibitor N-ω-Hydroxy-L norArginine (nor-NOHA) on high glucose cultured rhesus macaque retinal vascular endothelial cell line (RF/6A) in vitro.Methods The RF/6A cells were divided into the following 4 groups:normal control group (5.0 mmol/L of glucose,group A),high glucose group (25.0 mmol/L,group B),high glucose with 125 mg/L nor-NOHA group (group C),and high glucose with 1％ DMSO group (group D).The proliferation,migration ability and angiogenic ability of RF/6A cells were measured by Methyl thiazolyl tetrazolium (MTT),transwell chamber and tube assay respectively.The express of Arg Ⅰ,eNOS,iNOS mRNA of RF/6A cells were measured by real-time polymerase chain reaction (RT-PCR),Enzyme-linked immuno sorbent assay (ELISA) was used to detect the expression of NO and interleukine (IL)-1b of RF/6A cells.Results The proliferation,migration,and tube formation ability of group A (t=2.367,5.633,7.045;P＜0.05) and group C (t=5.260,6.952,8.875;P＜0.05)were significantly higher than group B.RT-PCR results showed the Arg Ⅰ and iNOS expression in group B was higher than that in group A (t=6.836,3.342;P＜0.05) and group C (t=4.904,7.192;P＜0.05).The eNOS expression in group B was lower than that in group A and group C (t=4.165,6.594;P＜0.05).ELISA results showed NO expression in group B was lower than that in group A and group C (t=4.925,5.368;P＜0.05).IL-1b expression in group B was higher than that in group A and group C (t=5.032,7.792;P＜0.05).Conclusions Nor-NOHA has a protective effect on cultured RF/6A cells in vitro and can enhance its proliferation,migration and tube formation.The mechanism may be inhibiting the oxidative stress by balancing the expression of Arg/NOS.

Objective To investigate the effects of exosomes from cultured human retinal pigment epithelium (ARPE-19) cells affected by oxidative stress on the proliferation and expression of vascular endothelial growth factor-A (VEGF-A) and Akt of ARPE-19 cells. Methods Culture ARPE-19 cells. The concentration of 2.5μmol/L rotenone was selected to simulate oxidative stress and isolated ARPE-19-exosome. Exosomes were isolated by ExoQuick exosome precipitation solution. Transmission electron microscopy was used to identify the morphology of exosomes. Western blot was used to detect exosomes’ surface-specific maker protein CD63. ARPE-19 cells affected by oxidative stress were cultured with exosome as experimental group, normal ARPE-19 cells were cultured with exosome as control group. The cell proliferation was examined by methyl thiazolyl tetrazolium assay. Western blot and immunofluorescence assay were used to detect the expression levels of VEGF-A and Akt protein. Real-time quantitative polymerase chain reaction (RT-PCR) was used to detect the levels of VEGF-A mRNA and Akt mRNA. Results The diameter of normal ARPE-19-exosomes ranged from 50 to 150 nm. The isolated exosomes expressed CD63. AREP-19 cells were cultured with ARPE-19 (affected by rotenone)-exosome, the cell viability in experimental group was significantly reduced than in the control group. Green fluorescence was observed in the cytoplasm under fluorescence microscope. Compared with the control group, VEGF-A was up-regulated expressed and Akt was down-regulated expressed. Western blot results showed that, VEGF-A protein expression in the experimental group were higher than the control group. Akt protein expression in the experimental group were less than the control group. The difference was statically significant (t=3.822, 6.527;P<0.05). RT-PCR results showed that VEGF-A mRNA expression levels was higher in the experimental group than the control group. Akt mRNA expression levels was lower in the experimental group than the control group. The difference was statically significant (t=8.805,?7.823;P<0.05). Conclusions Exosomes from ARPE-19 cells affected by oxidative stress inhibit the proliferation of normal ARPE-19 cells, increase the expression of VEGF-A and reduce the expression of Akt.