Introduction: The contribution of the family environment to childhood obesity in Malaysia is not well known. This paper describes the study, methodology and results of a pilot study to assess the feasibility of conducting a study on diet and lifestyle factors among Malay primary school children and their main caregiver(s) in regard to body weight status. Methods: The Family Diet Study used a cross-sectional design and targeted a minimum of 200 Malay families at five national primary schools in the Klang Valley, Malaysia using a multi-stage sampling method. Participants were Malay families with children aged 8 to 12 years and their main caregiver(s). Data on socio-demographic, dietary intake, parental child feeding practices, physical activity and anthropometric measures were collected predominantly at schools with follow-up 24-h dietary recalls collected by phone. Details of recruitment, inclusion criteria, assessments and statistical analyses are also discussed. Results: Eleven families provided data by answering questionnaires, recalling diet intake and participating in anthropometric measures. The results showed overall feasibility of the study protocol but required some modifications prior to implementation of the main study. Mothers were the main parent involved in family food procurement, preparation and mealtime supervision. Snacking was not commonly reported and fruit and vegetables intakes were generally infrequent. Conclusion: The most novel component of this study was the comprehensive collection of data from both children and their main caregiver(s) within the context of the family. Detailed information on dietary and lifestyle aspects will help to elucidate factors associated with obesity aetiology in Malay children.

OBJECTIVETo investigate the effects of microRNA-133a on isoproterenol (ISO)-induced neonatal rat cardiomyocyte hypertrophy and related molecular mechanism focusing on the changes of L-type calcium channel α1C subunit.

METHODSNeonatal rat cardiomyocytes were cultured, cardiomyocyte hypertrophy was induced by isoproterenol (ISO, 10 µmol/L). The cell surface area was measured by phase contrast microscope and Leica image analysis system. The mRNA expressions of atrial natriuretic peptide (ANP), β-myosin heavy chain (β-MHC), miR-133a and the α1C were detected by qRT-PCR. The protein expression of α1C was evaluated by Western blot. MiR-133a mimic was transfected into cardiomyocytes to investigate the effects of miR-133a on ISO-induced cardiomyocyte hypertrophy. The targets of miR-133a were predicted by online database Targetscan. The 3' untranslated region sequence of α1C was cloned into luciferase reporter vector and then transiently transfected into HEK293 cells. The luciferase activities of samples were measured to verify the expression of luciferase reporter vector. The expression level of α1C was inhibited by RNAi to determine the effects of α1C on cardiomyocyte hypertrophy. Intracellular Ca(2+) content was measured by confocal laser microscope.

RESULTS(1) The expression of miR-133a was significantly reduced in ISO-induced cardiomyocyte hypertrophy (P < 0.01) . Upregulating miR-133a level could suppress the increase of cell surface area, the mRNA expression of ANP and β-MHC (P < 0.01) . (2) α1C was the one of potential target of miR-133a by prediction using online database Targetscan. The luciferase activities of HEK293 cells with the plasmid containing wide type α1C 3'UTR sequence were significantly decreased compared with control group (P < 0.01) . Upregulation of the miR-133a level by miR-133a mimic transfection could suppress the protein expression of α1C (P < 0.05) . (3) The expression of α1C was significantly increased in ISO treated cardiomyocytes (P < 0.05) . Downregulation of α1C by RNAi could markedly inhibit the increase of cell surface area, the mRNA expression of ANP and β-MHC (P < 0.01, P < 0.05, P < 0.05). (4) Downregulation of α1C expression by RNAi or upregulation of miR-133a level by miR-133a mimic transfection significantly inhibited intracellular Ca(2+) content (P < 0.01) .

CONCLUSIONSOur data confirms that α1C is the target of miR-133a. MiR-133a can negatively regulate the expression of L-type calcium α1C subunit, resulting in the decrease of intracellular Ca(2+) content and the attenuation of ISO-induced cardiomyocyte hypertrophy.

Animals ; Calcium Channels, L-Type ; metabolism ; Cell Enlargement ; drug effects ; Cells, Cultured ; Isoproterenol ; pharmacology ; MicroRNAs ; genetics ; Myocytes, Cardiac ; drug effects ; metabolism ; pathology ; Rats ; Rats, Sprague-Dawley ; Transfection

OBJECTIVETo investigate the negative regulation of microRNA-1 (miR-1) on L-type calcium channel beta2 subunit (Cavbeta 2) during cardiomyocyte hypertrophy and its mechanism.

METHODSCardiomyocyte hypertrophy was induced by isoproterenol (ISO). The cell surface area was measured by image analysis system (HJ2000). The targets of miR-1 were predicted by online database microCosm. The 3' untranslated region sequence of Cavbeta 2 was cloned into luciferase reporter vector and then transiently transfected into HEK293 cells. The luciferase activities of samples were measured to verify the expression of luciferase reporter vector. The expression of atrial natriuretic peptide (ANP), beta-myosin heavy chain (beta-MHC), miR-1 and the Cavbeta 2 mRNA were detected by qRT-PCR. The protein expression of Cavbeta 2 was detected by Western blot. The level of miR-1 was up-regulated by miR-1 mimic transfection and the expression level of Cavbeta 2 was down-regulated by RNAi, then effects of which on cardiomyocyte hypertrophy were investigated.

RESULTS(1) The expression of miR-1 was significantly reduced in cardiomyocyte hypertrophy. Upregulating the miR-1 level could suppress the increase of cell surface area, the expression of ANP and beta-MHC mRNA (P < 0.05). (2) Cavbeta 2 was the one of potential targets of miR-1 by prediction using online database microCosm. The luciferase activities of HEK293 cells with the plasmid containing miR-1 and wide type Cavbeta 3' UTR sequence was significantly decreased when compared with that of control group (P < 0.01). Up-regulation of the miR-1 level could suppress the protein expression of Cavbeta 2. (3) The expression of Cavbeta 2 was significantly increased in cardiomyocyte hypertrophy induced by ISO. Downregulation of Cavbeta by RNAi could markedly inhibit the increase of cell surface area, the expression of ANP and beta-MHC mRNA.

CONCLUSIONCavbeta2 is one of potential targets of miR-1 by bioinformatics prediction. The experiment data confirms that Cavbeta2 is truly the target of miR-1. MiR-1 can negatively regulate the expression of Cavbeta 2, resulting in the decrease of intracellular Ca2+ content and the attenuation of cardiomyocyte hypertrophy.

Animals ; Atrial Natriuretic Factor ; metabolism ; Calcium Channels, L-Type ; genetics ; Cardiomegaly ; genetics ; Gene Expression Regulation ; HEK293 Cells ; Humans ; MicroRNAs ; genetics ; Rats ; Rats, Sprague-Dawley ; Transfection ; Ventricular Myosins ; metabolism

OBJECTIVETo investigate the protective effect of Crocin against hypoxia damage of cardiac myocytes of neonatal rats and the regulation of HIF-1 and prolyhydroxylase (PHDs).

METHODSA model of CoCl2 simulated hypoxia damage was established in primary cultural myocardial cell. Expression levels of HIF-1alpha, VEGF, iNOS, as well as PHD1, 2, 3 protein in myocardial cells were detected by Western blot.

RESULTSCompared with CoCl2 group, the viability of myocardial cell was significantly increased after treated 24 h at 10(-5)mol/L Crocin (P < 0.01), HIF-1alpha, VEGF and iNOS were expressed higher than those in Crocin + CoCl2 group (P < 0.01), the expression of PHD2 was significantly increased (P < 0.01), while the expression of PHD3 was remarkably reduced in Crocin + CoCl2 Group (P < 0.01).

CONCLUSIONCrocin has better protective effect on hypoxic damage of myocardial cell. The mechanisms of protective effect of Crocin may be related to the activation of HIF-1-mediated pathway of the hypoxia response. PHDs may be involved in the pathophysiology regulated process of myocardial cells.

Animals ; Carotenoids ; pharmacology ; Cell Hypoxia ; drug effects ; Cells, Cultured ; Homeodomain Proteins ; metabolism ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Hypoxia-Inducible Factor-Proline Dioxygenases ; metabolism ; Myocytes, Cardiac ; drug effects ; metabolism ; Nitric Oxide Synthase Type II ; metabolism ; Procollagen-Proline Dioxygenase ; metabolism ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo investigate the feasibility of establishing an integrated regional network for ST-segment elevation myocardial infarction (STEMI) care in China and evaluate the implementation effect of this network.

METHODSBased on real-time electrocardiogram transmission technology, we established an integrated regional network for STEMI care (IRN-STEMI) with Xiamen Heart Center as the core center, 120 Emergency Systems, PCI-capable hospitals and other community health units as core elements of this network. Reperfusion treatment data of Xiamen Heart Center including the number of patients receiving primary percutaneous coronary intervention (PCI), the mean first medical contact to balloon (FMC-to-B) time, the mean door to balloon (D-to-B) time, the mean length of hospital stay, the mean medical cost and in-hospital mortality were compared before (n = 165) and at 1 year after the built-up of IRN-STEMI (n = 343).

RESULTSCompared to pre-IRN-STEMI era, primary PCI ratio (84.5% (290/343) vs. 75.5% (185/245)) were significantly increased post establishment of IRN-STEMI within the network (P = 0.06). STEMI patients admitted in Xiamen Heart Center was significantly increased from 165 to 256, the annual mean FMC-to-B time ((110.3 ± 34.0)min vs. (137.9 ± 58.5) min, P < 0.01) and D-to-B ( (76.5 ± 33.0) min vs. (107.3 ± 38.0) min, P < 0.01) , as well as the mean medical cost were significantly decreased ( (51 398 ± 22 100) RMB vs. (56 970 ± 24 593) RMB, P < 0.05), while the mean length of hospital stay ((9.0 ± 4.3)d vs. (9.7 ± 4.8)d, P > 0.05) and in-hospital mortality (3.1% (8/256) vs. 3.0% (5/165) , P > 0.05) remained unchanged before and after the setting of IRN-STEMI in Xiamen Heart Center.

CONCLUSIONEstablishment of an integrated regional network system for STEMI patients in China is feasible. With collaboration of qualified heart center, EMS and PCI-capable and non-PCI capable local hospitals, establishment of IRN-STEMI effectively increased the ratio of primary PCI for STEMI patients, it also significantly shortened the FMC-to-B and D-to-B time, decreased mean medical cost, thus, the regional IRN-STEMI network might be an effective working system for improving the medical care for STEMI patients.

China ; epidemiology ; Community Networks ; Cost Control ; Electrocardiography ; Hospital Mortality ; Hospitalization ; Humans ; Length of Stay ; Myocardial Infarction ; mortality ; therapy ; Percutaneous Coronary Intervention ; Time Factors

The complex spatial and temporal organization of neural activity in the brain is important for information-processing that guides behavior. Hence, revealing the real-time neural dynamics in freely-moving animals is fundamental to elucidating brain function. Miniature fluorescence microscopes have been developed to fulfil this requirement. With the help of GRadient INdex (GRIN) lenses that relay optical images from deep brain regions to the surface, investigators can visualize neural activity during behavioral tasks in freely-moving animals. However, the application of GRIN lenses to deep brain imaging is severely limited by their availability. Here, we describe a protocol for GRIN lens coating that ensures successful long-term intravital imaging with commercially-available GRIN lenses.
Animals ; Biocompatible Materials ; Brain ; physiology ; Hippocampus ; cytology ; Lenses ; Mice, Inbred C57BL ; Mice, Transgenic ; Microscopy, Fluorescence ; methods ; Neuroimaging ; instrumentation ; methods ; Neurons ; physiology