Urotensin-II receptor stimulation of cardiac L-type Ca2+ channels requires the ßγ subunits of Gi/o-protein and phosphatidylinositol 3-kinase-dependent protein kinase C ß1 isoform.

Recent studies have demonstrated that urotensin-II (U-II) plays important roles in cardiovascular actions including cardiac positive inotropic effects and increasing cardiac output. However, the mechanisms underlying these effects of U-II in cardiomyocytes still remain unknown. We show by electrophysiological studies that U-II dose-dependently potentiates L-type Ca(2+) currents (ICa,L) in adult rat ventricular myocytes. This effect was U-II receptor (U-IIR)-dependent and was associated with a depolarizing shift in the voltage dependence of inactivation. Intracellular application of guanosine-5'-O-(2-thiodiphosphate) and pertussis toxin pretreatment both abolished the stimulatory effects of U-II. Dialysis of cells with the QEHA peptide, but not scrambled peptide SKEE, blocked the U-II-induced response. The phosphatidylinositol 3-kinase (PI3K) inhibitor wortmannin as well as the class I PI3K antagonist CH132799 blocked the U-II-induced ICa,L response. Protein kinase C antagonists calphostin C and chelerythrine chloride as well as dialysis of cells with 1,2bis(2aminophenoxy)ethaneN,N,N',N'-tetraacetic acid abolished the U-II-induced responses, whereas PKCα inhibition or PKA blockade had no effect. Exposure of ventricular myocytes to U-II markedly increased membrane PKCß1 expression, whereas inhibition of PKCß1 pharmacologically or by shRNA targeting abolished the U-II-induced ICa,L response. Functionally, we observed a significant increase in the amplitude of sarcomere shortening induced by U-II; blockade of U-IIR as well as PKCß inhibition abolished this effect, whereas Bay K8644 mimicked the U-II response. Taken together, our results indicate that U-II potentiates ICa,L through the ßγ subunits of Gi/o-protein and downstream activation of the class I PI3K-dependent PKCß1 isoform. This occurred via the activation of U-IIR and contributes to the positive inotropic effect on cardiomyocytes.

Nesfatin-1 Suppresses Cardiac L-type Ca²âº Channels Through Melanocortin Type 4 Receptor and the Novel Protein Kinase C Theta Isoform Pathway.

BACKGROUND/AIMS: Nesfatin-1 (NF-1), an anorexic nucleobindin-2 (NUCB2)-derived hypothalamic peptide, acts as a peripheral cardiac modulator and it can induce negative inotropic effects. However, the mechanisms underlying these effects in cardiomyocytes remain unclear. METHODS: Using patch clamp, protein kinase assays, and western blot analysis, we studied the effect of NF-1 on L-type Ca2+ currents (ICa,L) and to explore the regulatory mechanisms of this effect in adult ventricular myocytes. RESULTS: NF-1 reversibly decreased ICa,L in a dose-dependent manner. This effect was mediated by melanocortin 4 receptor (MC4-R) and was associated with a hyperpolarizing shift in the voltage-dependence of inactivation. Dialysis of cells with GDP-ß-S or anti-Gß antibody as well as pertussis toxin pretreatment abolished the inhibitory effects of NF-1 on ICa,L. Protein kinase C (PKC) antagonists abolished NF-1-induced responses, whereas inhibition of PKA activity or intracellular application of the fast Ca2+-chelator BAPTA elicited no such effects. Application of NF-1 increased membrane abundance of PKC theta isoform (PKCθ), and PKCθ inhibition abolished the decrease in ICa,L induced by NF-1. CONCLUSION: These data suggest that NF-1 suppresses L-type Ca2+ channels via the MC4-R that couples sequentially to the ßγ subunits of Gi/o-protein and the novel PKCθ isoform in adult ventricular myocytes.

The effectiveness of intervention in hepatitis C patients and improvement in their referral rate.

BACKGROUND: To investigate the effectiveness of the intervention with critical value management and push short messaging service (SMS), and to determine improvement in the referral rate of patients with positive hepatitis C antibody (anti-HCV). METHODS: No intervention was done for patients with positive anti-HCV screening results from 1 January 2015 to 31 October 2021. Patients with positive anti-HCV results at our hospital from 1 November 2021 to 31 July 2022 were informed vide critical value management and push SMS. For inpatients, a competent physician was requested to liaise with the infectious disease physician for consultation, and patients seen in the OPD (outpatient department) were asked to visit the liver disease clinic. The Chi-square correlation test, one-sided two-ratio test and linear regression were used to test the relationship between intervention and referral rate. RESULTS: A total of 638,308 cases were tested for anti-hepatitis C virus (HCV) in our hospital and 5983 of them were positive. 51.8% of the referred patients were aged 18-59 years and 10.8% were aged ≥75 years. The result of Chi-square correlation test between intervention and referral was p = .0000, p < .05. One-sided two-ratio test was performed for statistics of pre-intervention referral rate (p1) and post-intervention referral rate (p2). Normal approximation and Fisher's exact test for the results obtained were 0.000, p < .05, and the alternative hypothesis p1 - p2 < 0 was accepted. The linear regression equation was referral = 0.1396 × intervention + 0.3743, and the result model p = 8.79e - 09, p < .05. The model was significant, and the coefficient of intervention was 0.1396. CONCLUSIONS: The interventions of critical value management and push SMS were correlated with the referral rate of patients with positive anti-HCV.

Correlation Between the Number of Fiberoptic Bronchoscopies and Nosocomial Infection/Colonization of Carbapenem-Resistant Enterobacteriaceae.

Objective: The present study aims to explore the effects of different numbers of fiberoptic bronchoscopic examinations on the nosocomial infection/colonization of carbapenem-resistant Enterobacteriaceae (CRE). Methods: The data of 129 patients admitted to the intensive care unit of a grade 3A hospital were retrospectively analyzed, and CRE nosocomial infection/colonization situations in patients with fiberoptic bronchoscope application times of 1, 2, 3, and ≥4 were statistically analyzed. Results: The incidence of nosocomial infection/colonization of CRE increased significantly when the number of fiberoptic bronchoscopic examinations was ≥3. Conclusion: Nosocomial infection/colonization of CRE is highly correlated with an increased number of fiberoptic bronchoscopic examinations.

The effects of a short-term long-chain-triglyceride infusion on the postoperative immune function of pediatric patients receiving a gastrointestinal surgical procedure.

OBJECTIVE: This clinical trial investigates whether short-term administration of long chain triglycerides (LCT) has any influence on the immune function in children following gastrointestinal surgery. METHODS: Sixty pediatric patients receiving a gastrointestinal operation were randomly divided into the experimental group (n = 36) and the control group (n = 24). After abdominal operation, the subjects received parenteral nutrition (PN) support with or without LCT for 5 days. The patients' fasting blood samples were respectively collected at 24 hours preoperative, then 24 hours and 120 hours postoperative. Blood parameters related to the patients' immune function were measured. RESULTS: Before surgery and LCT treatment, the experimental group and control group did not differ significantly in overall state of health. Except for a small increase of serum IgM at 24 hours postsurgery (p < .05), all parameters representing the patients' immune function showed no significant difference between the LCT group and the control group with respect to peripheral blood mononuclear cell (PBMC), T lymphocyte, CD4, CD8, CD4/CD8, serum immunoglobulin A (IgA), IgG, IgM, complement C3, C4, interleukin (IL)-2, IL-4, IL-10, IL-12, tumor necrosis factor (TNF)-alpha and IFN-gamma (p > .05, respectively) before the operation, 24 hours and 120 hours after the operation. CONCLUSIONS: A short-term LCT administration at an appropriate dosage and infusion speed does not alter the pediatric patients' immune function after gastrointestinal surgery. The etiology and clinical significance of the slightly increased IgM 24 hours postsurgery need to be further investigated.

A short-term long-chain triglycerides infusion has no influence on immune function of adult patients undergoing gastrointestinal surgery.

BACKGROUND: Parenteral nutrition (PN) support containing long-chain triglycerides (LCT) plays a critical supportive role in surgical patients' management. This study aims to investigate the effects of intravenous (IV) LCT emulsion on human immune function in adult patients receiving a gastrointestinal surgical procedure. METHODS: Sixty adult patients were randomly assigned either to the LCT treatment group (n = 32) or to the control group (n = 28). After an abdominal operation, the subjects received PN treatment with or without LCT for 5 days. Neutrophil, peripheral blood mononuclear cell (PBMC), lymphocyte and CD4/CD8, serum immunoglobulin A (IgA), IgG, IgM, complement C3 and C4, interleukin (IL)-2, IL-4, IL-10, IL-12, tumor necrosis factor (TNF)-alpha, and interferon (IFN)-gamma were measured and statistically analyzed. RESULTS: The LCT and control groups did not differ significantly at entry in terms of general features. Except for a significant increase of neutrophil number at 24 hours after the surgery in both groups (p < .01), all parameters representing the patients' immune function had no significant difference between the LCT and the control groups with respect to neutrophil and PBMC count, lymphocyte, CD4/CD8, serum IgA, IgG, IgM, complement C3, C4, IL-2, IL-4, IL-10, IL-12, TNF-alpha, and IFN-gamma (p > .05, respectively) 24 hours before the operation, and 24 hours and 120 hours after the operation. CONCLUSIONS: The regimens of LCT administration may have diverse effects on human immune function in different patient populations. However, LCT emulsion at an appropriate dose and infusion speed does not alter human immune function of adult patients undergoing moderate gastrointestinal surgery.