κ Opioid Receptor Agonist Inhibits Myocardial Injury in Heart Failure Rats through Activating Nrf2/HO-1 Pathway and Regulating Ca2+-SERCA2a.

OBJECTIVES: We aimed to observe the protective effect of κ opioid receptor (κ-OR) agonist on myocardial injury in heart failure (HF) rats and its effect on Ca2+-SERCA2a and to explore the regulatory mechanism with the Nrf2/HO-1 signaling pathway. METHODS: 50 Sprague-Dawley rats were randomly divided into the following groups: the sham operation group (sham group), HF model group (HF group), HF+κ-OR agonist U50488 group (HU group), HF+U50488H+novel calmodulin-dependent protein kinase II (CaMKII) agonist (oleic acid) (HUO group), and HF+U50488H+Nrf2 inhibitor (HUM group). The HF rat's model was established through surgical ligation of the left anterior descending coronary artery and the exhausting swimming exercise. After that, rat's cardiac function was monitored by echocardiography. HE and MASSON staining was used to detect the myocardial injury, and TUNEL staining was used to detect the myocardial apoptosis. ELISA was performed to detect the biomarkers of oxidative stress. Moreover, the distribution of reactive oxygen species (ROS) and Nrf2 was detected under immunofluorescence. The expression of sarco/endoplasmic reticulum calcium (Ca2+) ATPase (SERCA) 2a, calmodulin, endoplasmic reticulum stress- (ERS-) related proteins, and Nrf2/HO-1 signaling pathway-related proteins were detected by Western Blotting. RESULTS: κ-OR agonist U50488H can significantly enhance rat's cardiac function, reduce the injury and apoptosis of myocardial cells, and alleviate endoplasmic reticulum stress injury in HF rats via upregulating the SERCA2a expression and inhibiting the Ca2+ influx. Furthermore, U50488H could also inhibit the phosphorylation of CaMKII and cAMP-response element binding protein (CREB). Additionally, administration of CaMKII-specific agonist could partially block the therapeutic effect of κ-OR agonist on the myocardium of HF rats. Interestingly, the antagonist of Nrf2 could also significantly reverse the therapeutic effect of κ-OR agonist. Therefore, these results suggested that the effect of U50488H on HF rats is dependent on regulating CaMKII phosphorylation and activating the Nrf2/HO-1 pathway. CONCLUSION: κ-OR agonists U50488H can improve ERS in cardiomyocytes and relieve myocardial injury in HF rats through activating the Nrf2/HO-1 pathway and regulating Ca2+-SERCA2a to inhibit Ca2+ influx.

IL-38 restrains inflammatory response of collagen-induced arthritis in rats via SIRT1/HIF-1α signaling pathway.

OBJECTIVE: To observe the restraining effect of IL-38 on inflammatory response in collagen-induced arthritis rats (CIA), and to explore the regulatory mechanism of SIRT1/HIF-1α signaling pathway. METHODS: 40 SD rats were randomly divided into Control group, CIA group, CLL group and CLH group, with 10 rats in each group; CIA rat model was established. The effects of IL-38 on arthritis index, inflammatory response, osteogenic factor and angiogenic factor were observed by methods including HE staining, ELISA, immunohistochemical and immunofluorescence. Human synoviocytes were cultured in vitro, and SIRT1 inhibitors were added to detect the expression for relating factors of SIRT1/HIF-1α signaling pathway by Western blot. RESULTS: IL-38 could alleviate CIA joint damage and restrain inflammatory response, could up-regulate the expression of OPG in CIA rats and could down-regulate the expression of RANKL and RANK. IL-38 could restrain the expression of VEGF, VEGFR1, VEGFR2 and HIF. Moreover, we found that IL-38 could up-regulate the SIRT1 expression and down-regulate the HIF-1α, TLR4 and NF-KB p65 expression in CLL and CLH groups. From the treatment of synoviocytes to simulate the CIA model and the treatment of SIRT1 inhibitors, we demonstrated that the inhibitory effect of IL-38 on inflammatory factors and regulation of SIRT1/HIF-1α signaling pathway-related proteins were inhibited. CONCLUSION: IL-38 can restrain the inflammatory response of CIA rats, can promote the expression of osteogenic factors, can inhibit neovascularization, and can alleviate joint damage in rats. The mechanism may be related to the regulation of SIRT1/HIF-1α signaling pathway.

The Majority of the Migrant Factory Workers of the Light Industry in Shenzhen, China May Be Physically Inactive.

Physical inactivity is a strong risk factor of non-communicable diseases (NCD). In China, there are 250 million migrant factory workers, who are susceptible to physical inactivity and hence NCD because of work nature and setting. With random stratified sampling, 807 such workers of the light industry were recruited in Shenzhen, China and completed a self-administered questionnaire with informed consent. The prevalence of inadequate physical activity (defined according to the World Health Organization's recommendation on level of moderate/vigorous physical activity) was 95.4%. Of all participants, 69.1% showed "a very low level of physical activity" (VLLPA), defined as ≤30 minutes of weekly moderate/vigorous physical activity, which was significantly associated with female sex (Odds ratio [OR]=1.65), lower education level (OR=0.10 to 0.33, primary education as the reference group) and married status (OR=0.63, single status as the reference group). Adjusted for these factors, perceived social support (Adjusted OR=0.87) was negatively associated with VLLPA, while job stress due to workload, which was significant in the univariate analysis (OR=0.98), became non-significant (p=0.184). Significant interaction between perceived social support and perceived job stress onto VLLPA was found (p=0.044), implying that the negative association between job stress and VLLPA, which might reflect a potential response to cope with stress by performing exercises, was stronger among those with weaker social support. The extremely low level of physical activity rings an alarm, as it implies high risk of NCD, and as there are no existing programs promoting physical activity in this group. Interventions need to take into account social support, potential coping to job stress, and structural factors of the factory setting, while involving factories' management.

Online High-throughput Mutagenesis Designer Using Scoring Matrix of Sequence-specific Endonucleases.

CRISPR Cas9 and other sequence-specific endonucleases are fundamental genome editors supporting gene knockout and gene therapy. A speedy and accurate computational allele designer is required for a high through-put gene mutagenesis pipeline using these new techniques. An automatic system, Cas9 online designer (COD), was created to screen Cas9 targets and off-targets, as well as to provide gene knockout and genotyping strategies. A gene knockout rat model was successfully created and genotyped under the direction of this online system confirming its ability to predict real targets and off-targets. Gene knockout strategies to mutate 72 rat cytochrome P450 genes were designed instantly by the system to demonstrate its high-throughput efficiency. Also, the system used an off-target scoring matrix which can be applied to any sequence-specific genome editing tools besides Cas9. The COD system (http://cas9.wicp.net) has established a speedy, accurate, flexible and high through-put computational gene knockout pipeline supporting the sequence-specific endonuclease induced mutagenesis.

The association between multimorbidity and poor adherence with cardiovascular medications.

Multimorbidity, defined as the presence of two or more chronic conditions, leads to a substantial public health burden. This study evaluated its association with adherence with cardiovascular medications in a Chinese population. A proportional stratified sampling was adopted to draw a representative sample of residents living in Henan Province, China. Interviewer-administered surveys were conducted by trained researchers. The outcomes included the number of chronic medical conditions, adherence with long-term medications (MMAS-8), and depressive symptoms (CESD-20). Binary logistic regression analysis was conducted to evaluate if medication adherence was associated with the presence of multimorbidity. From a total of 3866 completed surveys, the proportion of subjects having 0, 1 and ≥2 chronic conditions was 62.6%, 23.8% and 13.5%, respectively. Among 27.6% who were taking chronic medications, 66.6% had poor medication adherence (MMAS-8 score≤6). From binary logistic regression analysis, subjects with poor medication adherence were significantly associated with multimorbidity (adjusted odds ratio [AOR]: 1.35, 95% C.I. 1.02-1.78, p=0.037). Other associated factors included older age (AOR=1.04, 95% C.I. 1.03-1.05, p<0.001), smoking (AOR=1.63, 95% C.I. 1.16-2.30, p=0.005), family history of hypertension (AOR=1.51, 95% C.I. 1.19-1.93, p=0.001), and fair to poor self-perceived health status (AOR=2.15, 95% C.I. 1.69-2.74, p<0.001). Using medication adherence as the outcome variable, multimorbidity was significantly associated with poor drug adherence (AOR=1.34, 95% C.I. 1.02-1.77, p=0.037). Multimorbidity was associated with poorer medication adherence. This implies the need for closer monitoring of the medication taking behavior among those with multiple chronic conditions.

Testis-specific Fank1 gene in knockdown mice produces oligospermia via apoptosis.

Fank1 is exclusively expressed in the testis from the meiosis phase to the haploid phase of spermatogenesis. In this study, we examined the function of Fank1 by establishing a Fank1-knockdown transgenic mouse model. The apoptotic statuses of the testes of the transgenic mice were tested using the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) method. The FANK1 consensus DNA-binding sequence was identified using cyclic amplification of sequence target (CAST) analysis. Differentially expressed genes were examined using microarray analysis. A reduction in sperm number and an increase in apoptotic spermatocytes were observed in Fank1-knockdown mice, and the apoptotic cells were found to be primarily spermatogonia and spermatocytes. The CAST results demonstrated that the consensus DNA-binding sequence was AAAAAG, in which the percentage occurrence of each base at each position ranged from 55 to 86%. This sequence was present in the promoter regions of 10 differentially expressed genes that were examined using microarray analysis. In total, 17 genes were differentially expressed with changes in their expression levels greater than twofold. The abnormal expression of Fank1 target genes that were regulated directly or indirectly by Fank1 reduced the number of sperm in the knockdown mice. Thus, FANK1 may play a pivotal role in spermatogenesis as a transcription factor.

Cloning and characterization of an apolipoprotein C2 promoter in the mouse central nervous system.

Apolipoprotein C2 is an important member of the apolipoprotein C family, and is a potent activator of lipoprotein lipase. In the central nervous system, apolipoprotein C2 plays an important role in the catabolism of triglyceride-rich lipoproteins. Studies into the exact regulatory mechanism of mouse apolipoprotein C2 expression have not been reported. In this study, seven luciferase expression vectors, which contained potential mouse apolipoprotein C2 gene promoters, were constructed and co-transfected with pRL-TK into HEK293T cells to investigate apolipoprotein C2 promoter activity. Luciferase assays indicated that the apolipoprotein C2 promoter region was mainly located in the +104 bp to +470 bp region. The activity of the different lengths of apolipoprotein C2 promoter region varied. This staggered negative-positive-negative arrangement indicates the complex regulation of apolipoprotein C2 expression and provides important clues for elucidating the regulatory mechanism of apolipoprotein C2 gene transcription.