RESUMO

BACKGROUND: The relationship between long-term outcomes and operator experience for left atrial appendage occlusion (LAAO) is still unknown. OBJECTIVES: This study sought to explore the association between operator LAAO experience and one-year clinical outcomes. METHODS: The RECORD study (Registry to Evaluate Chinese Real-World Clinical Outcomes in Patients With AF Using the WATCHMAN Left Atrial Appendage Closure Technology; NCT03917563) was a multicenter, prospective registry that included patients with the WATCHMAN LAAO device (Boston Scientific) in China from April 1, 2019, to October 31, 2020. The current analyses included patients with solely LAAO from the registry; those who had concomitant LAAO and ablation/other procedures were excluded. The primary outcome was a composite endpoint of death, stroke, systemic embolism, and Bleeding Academic Research Consortium (BARC)-defined type 3 or 5 bleeding at 1 year. RESULTS: A total of 1,547 LAAO patients and 111 operators were included. The mean ± SD CHA2DS2-VASc and HAS-BLED scores of patients were 4.0 ± 1.8 and 2.5 ± 1.1, respectively. The mean ± SD age of operators was 47.0 ± 7.2 years, 15 (13.5%) were female, and 52 (46.8%) were electrophysiologists. Utilizing maximally selected log-rank statistics, the thresholds to categorize an experienced operator were performing ≥32 LAAOs annually or ≥134 LAAOs in total. Performing ≥32 LAAOs annually is the better criterion than ≥134 LAAOs in total (absolute net reclassification index: 25.79%; P < 0.001). Compared with the ≥32 LAAO annually group, the <32 group was associated with a 1.8-fold (HRadjusted: 1.79; 95% CI: 1.16-2.78; P = 0.009) increase in the risk of the primary endpoint, and such risk in the <32 group can be reduced by ∼12% after performing each additional 5 cases (HRadjusted per 5 cases: 0.88; 95% CI: 0.78-0.99; P = 0.033). CONCLUSIONS: Performing ≥32 LAAOs annually could be a threshold to categorize an experienced operator. Before reaching this threshold, the risk of death, stroke, systemic embolism, and BARC-defined type 3 or 5 bleeding decreased by 12% after every 5 cases performed.

RESUMO

INTRODUCTION: Few studies have focused on the safety and efficacy of radiofrequency ablation (RFA) in treating incompetent great saphenous vein (GSV) in aged population. This study was designed to investigate the clinical efficacy of RFA in treating incompetent GSV in the aged patients. METHODS: In this retrospective study, we included 138 consecutive patients (involving 194 limbs) with a mean age of 63.0 years who underwent RFA and microphlebectomy or sclerotherapy due to symptomatic incompetent GSV with saphenofemoral junction reflux. Based on their ages, patients were classified into young group and aged group. Then we compared the preoperative and postoperative Clinical, Etiology, Anatomic, Pathophysiology (CEAP) classification, venous clinical severity score (VCSS) and chronic venous insufficiency questionnaire 14 (CIVIQ-14) score between the 2 groups. RESULTS: In both the young and aged groups, patients underwent RFA showed significant decrease in the CEAP and VCSS at month 1, 3 and 6 compared with immediately after RFA (month 0) (all P < .001). In addition, in both groups, significant increase was seen in the CIVIQ-14 score at month 1, 3 and 6 compared with month 0 (all P < .001). Compared with the young group, the post-RFA CEAP, VCSS and CIVIQ-14 scores showed no statistical differences in the aged group at the designated time points, respectively (all P > .05). CONCLUSIONS: RFA of GSV was effective for treating GSV in the aged population, which improved the CEAP, VCSS and CIVIQ-14.

RESUMO

INTRODUCTION: Immune checkpoint inhibitor-associated pneumonitis (CIP) is the most common immune-related advanced event (irAE). However, the risk factors of CIP occurrence and its relationship with prognosis remain to be clarified. This study aimed to explore biomarkers, prognosis, and efficacy of CIP occurrence in non-small cell lung cancer (NSCLC) patients who received anti-PD-1 inhibitors. METHODS: We performed a retrospective study in eligible NSCLC patients treated with anti-PD-1 inhibitors in Ruijin hospital. The receiver operating characteristic (ROC) curve and logistic regression were used for the optional cut-off value and the risk of CIP, respectively. The Kaplan-Meier method and Cox hazards regression models were used for survival analyses in CIP and non-CIP groups. RESULTS: Our study enrolled 229 patients, of which 35 (15.3 %) experienced CIP. CIP patients had higher proportions of male, current and former smoking, and history of pre-existing lung diseases. CIP patients also had a higher level of WBC (p = 0.025), ANC (p = 0.020), AEC (p = 0.025), and proportion of CD4+ T lymphocytes (p = 0.033) than those in non-CIP patients. Then patients were divided into two groups according to the cutoff value. It showed high baseline proportion of CD4+ T lymphocytes (OR = 4.027 (1.279-12.677), P = 0.017) and AEC (OR = 2.697 (1.047-6.945, P = 0.040) were independent predictors of CIP occurrence. CIP occurrence was an independent predictor of progression-free survival (PFS) in the enrolled patients. Regarding patient efficacy, severe-CIP patients had the highest ORR, followed by grade 1-2 CIP patients, and non-CIP patients (44.44 %, 35.3 %, and 28.35 %, respectively). CONCLUSION: The onset time of CIP occurrence was early in severe CIP patients, suggesting the importance of early identification and timely intervention of CIP. Baseline proportion of CD4+ T lymphocytes and AEC were independent predictors of CIP occurrence. In addition, CIP occurrence predicted higher ORR, longer PFS, and more opportunities for long-term survival benefits.

RESUMO

Human cytomegalovirus (HCMV) glycoprotein B (gB) is a class III membrane fusion protein required for viral entry. HCMV vaccine candidates containing gB have demonstrated moderate clinical efficacy, but no HCMV vaccine has been approved. Here, we used structure-based design to identify and characterize amino acid substitutions that stabilize gB in its metastable prefusion conformation. One variant containing two engineered interprotomer disulfide bonds and two cavity-filling substitutions (gB-C7), displayed increased expression and thermostability. A 2.8 Å resolution cryoelectron microscopy structure shows that gB-C7 adopts a prefusion-like conformation, revealing additional structural elements at the membrane-distal apex. Unlike previous observations for several class I viral fusion proteins, mice immunized with postfusion or prefusion-stabilized forms of soluble gB protein displayed similar neutralizing antibody titers, here specifically against an HCMV laboratory strain on fibroblasts. Collectively, these results identify initial strategies to stabilize class III viral fusion proteins and provide tools to probe gB-directed antibody responses.

Assuntos

RESUMO

A lack of access to handwashing facilities is a significant risk factor for lower respiratory infections(LRIs). However, no studies have reported epidemiologic changes in the burden of LRIs attributed to the lack of access to handwashing facilities. We conducted an integrated assessment of the burden of LRIs attributable to the lack of handwashing facilities from 1990 to 2019 using data from the Global Burden of Disease Study 2019. In 2019, 270,000 deaths were attributed to LRIs due to a lack of access to handwashing facilities, with DALYs reaching 14.02 million. The age-standardized mortality rate (ASMR) of LRIs caused by a lack of access to handwashing facilities was approximately 3.74, while the age-standardized DALY rate (ASDR) was reported to be 203.55 in 2019. Over the past 30 years, the burden of LRIs attributed to the lack of access to handwashing facilities has shown a global decline. In 2019, this burden was most pronounced in infants under 1 year of age and in those older than 95 years, reflecting the highest DALY (5591.83) and mortality rates (79.43), respectively. The burden of LRIs caused by the lack of access to handwashing facilities was found to be more severe in males and significantly more pronounced in regions with a low sociodemographic index (SDI), such as the Sahara African region. The development of targeted strategies to address the inadequate and unequal distribution of handwashing facilities holds important value in improving the disease burden of LRIs.

RESUMO

Owing to patient-derived tumor tissues and cells, significant advances have been made in personalized cancer treatment and precision medicine, with cancer stem cell-derived three-dimensional tumor organoids serving as crucial in vitro models that accurately replicate the structural, phenotypic, and genetic characteristics of tumors. However, despite their extensive use in drug testing, genome editing, and transplantation for facilitating personalized treatment approaches in clinical practice, the inadequate capacity of these organoids to effectively model immune cells and stromal components within the tumor microenvironment limits their potential. Additionally, effective clinical immunotherapy has led the tumor immune microenvironment to garner considerable attention, increasing the demand for simulating patient-specific tumor-immune interactions. Consequently, co-culture techniques integrating tumor organoids with immune cells and tumor microenvironment constituents have been developed to expand the possibilities for personalized drug response investigations, with recent advancements enhancing the understanding of the strengths, limitations, and applicability of the co-culture approach. Herein, the recent advancements in the field of tumor organoids have been comprehensively reviewed, specifically highlighting the tumor organoid co-culture-related developments with various immune cell models and their implications for clinical research. Furthermore, this review delineates the current state of research and application of organoid models regarding the therapeutic approaches and related challenges for gynecological tumors. This study may provide a theoretical basis for further research on the use of patient-derived organoids in tumor immunity, drug development, and precision medicine.

Assuntos

RESUMO

The differentiation of vascular endothelial cells and the formation of new blood vessels are inseparable from the energy supply and regulation of metabolism. The budding of blood vessels is a starting point of glycolysis pathway in angiogenesis. Phosphofructokinase-2/fructose 2,6-biophosphatase 3 (PFKFB3), a key rate-limiting enzyme in glycolysis, exhibits strong kinase activity. Inhibition of PFKFB3 can reduce the rate of glycolysis, thereby inhibiting the budding of blood vessels, resulting in inhibition of pathological angiogenesis. In this review, the role of PFKFB3 in the angiogenesis of inflammatory diseases was summarized, and the endothelial inflammatory diseases associated with PFKFB3 were reviewed.

RESUMO

Analyzing heart sound signals presents a novel approach for early diagnosis of pediatric congenital heart disease. The existing segmentation algorithms have limitations in accurately distinguishing the first (S1) and second (S2) heart sounds, limiting the diagnostic utility of cardiac cycle data for pediatric pathology assessment. This study proposes a time bidirectional long short-term memory network (TBLSTM) based on multi-scale analysis to segment pediatric heart sound signals according to different cardiac cycles. Mel frequency cepstral coefficients and dynamic characteristics of the heart sound fragments were extracted and input into random forest for multi-classification of congenital heart disease. The segmentation model achieved an overall F1 score of 94.15% on the verification set, with specific F1 scores of 90.25% for S1 and 86.04% for S2. In a situation where the number of cardiac cycles in the heart sound fragments was set to six, the results for multi-classification achieved stabilization. The performance metrics for this configuration were as follows: accuracy of 94.43%, sensitivity of 95.58%, and an F1 score of 94.51%. Furthermore, the segmentation model demonstrates robustness in accurately segmenting pediatric heart sound signals across different heart rates and in the presence of noise. Notably, the number of cardiac cycles in heart sound fragments directly impacts the multi-classification of these heart sound signals.

RESUMO

In addressing the critical challenges posed by the misuse and inefficiency of traditional pesticides, we introduce a Nano-Cocrystal material composed of the herbicide clopyralid and coformer phenazine. Developed through synergistic supramolecular self-assembly and mechanochemical nanotechnology, this Nano-Cocrystal significantly enhances pesticide performance. It exhibits a marked improvement in stability, with reductions in hygroscopicity and volatility by approximately 38%. Moreover, it intelligently modulates release according to environmental factors, such as temperature, pH, and soil inorganic salts, demonstrating decreased solubility by up to four times and improved wettability and adhesion on leaf surfaces. Importantly, the herbicidal activity surpasses that of pure clopyralid, increasing suppression rates of Medicago sativa L. and Oxalis corniculata L. by up to 27% at the highest dosage. This Nano-Cocrystal also shows enhanced crop safety and reduced genotoxicity compared to conventional formulations. Offering a blend of simplicity, cost-effectiveness, and robust stability, our findings contribute a sustainable solution to agricultural practices, favoring the safety of nontarget organisms.

Assuntos

RESUMO

OBJECTIVE: Morusin (Mor), a prenylated flavonoid isolated from the root bark of Morus alba L., exhibits potent anti-tumour effects; however, the molecular target of Mor is still not entirely clear. This study aimed to elucidate the mechanism of Mor against hepatocellular carcinoma (HCC) and identify potential molecular targets. METHODS: Mitochondrial function was assessed by measuring the mitochondrial membrane potential, mitochondrial ultrastructure, oxygen consumption, and ATP levels. Mor-induced mitophagy was confirmed using western blotting, immunofluorescence, and fluorescent probes. Transcriptomics, flow cytometry, western blotting, qRT-PCR and biochemical assays were used to reveal the molecular mechanisms and targets of Mor against HCC. We further validated the interaction between Mor and the target proteins using molecular docking and biolayer interferometry (BLI). The inhibitory effect of Mor in vivo was evaluated using a Hep3B murine xenograft model. RESULTS: Mor significantly reduced the ATP citrate lyase (ACLY) expression and inhibited ACLY activity in HCC cells. BLI analysis demonstrated a direct interaction between Mor and the ACLY active domain. Mor-induced ACLY inhibition led to ROS accumulation in HCC cells, which caused mitochondrial damage, triggered PINK1/Parkin-mediated mitophagy, and ultimately induced mitochondrial apoptosis. We further verified that ROS is crucial in the apoptotic action of Mor through experiments regarding an ROS scavenger. Mor also significantly inhibited tumour xenograft growth in vivo. In addition, analysis of human liver cancer clinical samples revealed elevated ACLY levels positively correlated with histologic grade. CONCLUSION: Collectively, our findings highlight Mor as a potent bioactive inhibitor of ACLY and a promising candidate for HCC therapy.

RESUMO

KEY MESSAGE: Here, we systematically analyzed the potential fusion and fission events of neighboring genes in Arabidopsis genome and analyzed the influence on the protein targeting.

Assuntos

RESUMO

To understand the structure of the plankton community and the ecological niche characteristics of their dominant species, sampling surveys of plankton were conducted in Baiyangdian Lake in the spring （March）, summer （July）, and autumn （September） of 2022. The changes in the plankton community during the three seasons were analyzed by constructing ecological network diagrams, non-metric multidimensional scaling analysis （NMDS）, and the ecological niche width. The niche overlap of zooplankton dominant species was evaluated by the improved Levins' formula and Petraitis' index. The interspecific connectivity of dominant species was judged using the chi-square test and interspecies connectivity coefficients. The results showed that the niche width of plankton in the whole area was low. Zooplankton was dominated by rotifers, and phytoplankton was dominated by diatoms, cyanobacteria, and green algae. There were significant seasonal changes in the community structures of plankton. Compared with that in summer and autumn, there were fewer species of plankton in spring and lower interspecies connectivity. The overlap of dominant species of zooplankton was high in summer, and the interspecific competition was intensified, whereas the interspecific overlap of phytoplankton was at a low level in all three seasons. There was a significant positive correlation （W > χ20.05） between phytoplankton in summer and autumn, and the community structure was stable. The interdomain ecological network of zooplankton and phytoplankton showed a high negative correlation ratio in autumn, especially between copepods and cladoceras of zooplankton and chlorophyta and cyanophyta of phytoplankton. The plankton species in Baiyangdian Lake were abundant, with obvious seasonal differences. The dominant species were mainly a narrow ecological niche. The plankton community was generally in a stable state, and there was a strong predation relationship between copepods and cladoceras and green algae and cyanobacteria.

Assuntos

RESUMO

Ethnopharmacological relevance: Vitamin D (VD), selenium preparations (Se), and thyroid hormone replacement therapy are commonly used to treat Hashimoto thyroiditis (HT). Increasing evidence suggests that traditional Chinese medicine (TCM) is an effective therapeutic strategy in the treatment of HT. Aim of the study: This study aimed to investigate the efficacy and safety of commonly-used drugs for HT. Materials and methods: A literature search was performed using PubMed, Web of Science, Cochrane Library, EMBASE, Chinese China National Knowledge Infrastructure (CNKI), Clinical Trial Registry (Chi CTR), China Science and Technology Journal Database (the VIP), Wanfang Database, and China Chinese Biomedical Database (CBM) from January 1, 2003, to December 31, 2022. The outcomes included TPOAb, TgAb, TSH, FT3, FT4, and adverse events. Our study was registered in PROSPERO (CRD42023449705). Results: Sixty trials and 4719 participants were included, comparing 16 treatments: VD, Se, LT-4, Se + LT-4, HM, placebo + LT-4, HM + LT-4, Se + myolnositol, Se + VD, HM + Se, mannan peptide, LT-4+prednisone, Methimazole, Methimazole + HM, Tapazole + Propranolol, and placebo. We found that Chinese herbal medicine has significant effect vs. LT-4 [MD 0.10, 95 % confidence interval 0.02 to 0.50]) and LT-4+placebo [MD 0.10, 95 % confidence interval 0.01 to 0.77]) in reducing TPOAb. Although receiving LT-4+prednisone was not statistically significant, the treatment ranking showed that this combination therapy had the highest probability of reducing TPOAb levels (72.8 %). In addition, the effect of Se plus LT-4 was not statistically significant; however, the treatment ranking showed that this combination therapy had the highest probability (78.6 %) of reducing TgAb levels, followed by HM (64.0 %). Reports on side effects have mainly focused on the digestive and cardiovascular systems. Conclusion: Our analyses showed that HM alone or in combination with other treatments for patients with HT can improve the side effects of other drugs, enhance efficacy, and maybe the most effective option for treating HT. However, there still need further verified using high-quality evidence.

RESUMO

Microplastics are a growing concern as pollutants that impact both public health and the environment. However, the toxic effects of polypropylene microplastics (PP-MPs) are not well understood. This study aimed to investigate the effects of PP-MPs on cardiotoxicity and its underlying mechanisms. The cardiotoxicity of exposure to different amounts of PP-MPs were investigated in both ICR mice and H9C2 cells. Our results demonstrated that sub-chronic exposure to 5 and 50 mg/L PP-MPs led to myocardial structural damage, apoptosis, and fibrosis in mice cardiomyocytes. Flow cytometry analysis revealed that PP-MPs could decrease mitochondrial membrane potential and induce apoptosis in H9C2 cells. Western blotting revealed decreased expression of Bcl-2, poly(ADP-ribose) polymerase (PARP) and caspase 3 and increased expression of Bax, cleaved-PARP, and cleaved-caspase 3 in PP-MPs-treated cardiac tissue and H9C2 cells. These results confirmed the apoptotic effects induced by PP-MPs. Moreover, PP-MPs treatment triggered oxidative stress, as evidenced by the increased levels of malondialdehyde; reduction in glutathione peroxidase, superoxide dismutase, and catalase activities in mice cardiac tissues; and increased reactive oxygen species levels in H9C2 cells. Finally, western blotting demonstrated that exposure to PP-MPs significantly reduced the expression levels of Nrf2 and p-ERK proteins associated with MAPK-Nrf2 pathway in both cardiac tissue and H9C2 cells. Overall, our findings indicate that PP-MPs can induce cardiomyocyte apoptosis through MAPK-Nrf2 signaling pathway, which is triggered by oxidative stress. This study provides a foundation for determining the effects of PP-MPs on cardiotoxicity and their underlying mechanisms.

RESUMO

Harmine (HM), a ß-carboline alkaloid extracted from plants, is a crucial component of traditional Chinese medicine (TCM) known for its diverse pharmacological activities. Thrombocytopenia, a common and challenging hematological disorder, often coexists with serious illnesses. Previous research has shown a correlation between HM and thrombocytopenia, but the mechanism needs further elucidation. The aim of this study was to clarify the mechanisms underlying the effects of HM on thrombocytopenia and to develop new therapeutic strategies. Flow cytometry, Giemsa staining, and Phalloidin staining were used to assess HM's impact on Meg-01 and HEL cell differentiation and maturation in vitro. A radiation-induced thrombocytopenic mouse model was employed to evaluate HM's effect on platelet production in vivo. Network pharmacology, molecular docking, and protein blotting were utilized to investigate HM's targets and mechanisms. The results demonstrated that HM dose-dependently promoted Meg-01 and HEL cell differentiation and maturation in vitro and restored platelet levels in irradiated mice in vivo. Subsequently, HM was found to be involved in the biological process of platelet production by upregulating the expressions of Rac1, Cdc42, JNK, and 5-HTR2A. Furthermore, the targeting of HM to 5-HTR2A and its correlation with downstream Rac1/Cdc42/JNK were also confirmed. In conclusion, HM regulates megakaryocyte differentiation and thrombopoiesis through the 5-HTR2A and Rac1/Cdc42/JNK pathways, providing a potential treatment strategy for thrombocytopenia.

RESUMO

Sorghum is an important C4 crop with various food and nonfood uses. Although improvements through hybridization and selection have been exploited, the introduction of genetic variation and the development of new genotypes in sorghum are still limited. Fast-neutron (FN) mutagenesis is a very effective method for gene functional studies and to create genetic variability. However, the full spectrum of FN-induced mutations in sorghum is poorly understood. To address this, we generated an FN-induced mutant population from the inbred line 'BTx623' and sequenced 40 M1 seedlings to evaluate the mutagenic effects of FNs on sorghum. The results show that each line had an average of 43.7 single-base substitutions (SBSs), 3.7 InDels and 35.15 structural variations (SVs). SBSs accounted for approximately 90.0% of the total number of small mutations. Among the eight treatment groups, FN irradiation at a dose of 19 Gy generated the highest number of mutations. The ratio of transition/transversion ranged from 1.77 to 2.21, and the G/C to A/T transition was the most common substitution in all mutant lines. The distributions of the identified SBSs and InDels were similar and uneven across the genome. An average of 3.63 genes were mutated in each mutant line, indicating that FN irradiation resulted in a suitable density of mutated genes, which can be advantageous for improving elite material for one specific or a few traits. These results provide a basis for the selection of the suitable dose of mutagen and new genetic resources for sorghum breeding.

Assuntos

RESUMO

BACKGROUND: Vancomycin-induced acute kidney injury (VI-AKI) is one of its serious adverse reactions. The purpose of this study is to discuss the risk factors for VI-AKI in overweight patients and construct a clinical prediction model based on the results of the analysis. METHODS: Multivariable logistic regression analysis was used to identify risk factors for VI-AKI and constructed nomogram models. The performance of the nomogram was evaluated based on the area under the receiver operating characteristic curve (AUC), calibration curves, and decision curve analysis (DCA). RESULT: Cancer (OR 4.186, 95% CI 1.473-11.896), vancomycin trough concentration >20.0 µg/mL (OR 6.251, 95% CI 2.275-17.180), concomitant furosemide (OR 2.722, 95% CI 1.071-6.919) and vasoactive agent (OR 2.824, 95% CI 1.086-7.340) were independent risk factors for VI-AKI. The AUC of the nomogram validation cohorts were 0.807 (95% CI 0.785-0.846). The calibration curve revealed that the predicted outcome was in agreement with the actual observations. Finally, the DCA curves showed that the nomogram had a good clinical applicability value. CONCLUSION: There are four independent risk factors for the occurrence of VI-AKI in overweight patients, and the nomogram prediction model has good predictive ability, which can provide reference for clinical decision-making.

RESUMO

The full-length prefusion-stabilized SARS-CoV-2 spike (S) is the principal antigen of COVID-19 vaccines. Vaccine efficacy has been impacted by emerging variants of concern that accumulate most of the sequence modifications in the immunodominant S1 subunit. S2, in contrast, is the most evolutionarily conserved region of the spike and can elicit broadly neutralizing and protective antibodies. Yet, S2's usage as an alternative vaccine strategy is hampered by its general instability. Here, we use a simulation-driven approach to design S2-only immunogens stabilized in a closed prefusion conformation. Molecular simulations provide a mechanistic characterization of the S2 trimer's opening, informing the design of tryptophan substitutions that impart kinetic and thermodynamic stabilization. Structural characterization via cryo-EM shows the molecular basis of S2 stabilization in the closed prefusion conformation. Informed by molecular simulations and corroborated by experiments, we report an engineered S2 immunogen that exhibits increased protein expression, superior thermostability, and preserved immunogenicity against sarbecoviruses.

Assuntos

RESUMO

BACKGROUND: Immune response and inflammation play important roles in the physiological and pathophysiological processes of heart failure (HF). In our previous study, myeloid-derived suppressor cells (MDSCs), a heterogeneous group of immature myeloid cells with anti-inflammatory and immunosuppressive functions, were shown to exert cardioprotective effects in HF. The pharmacological targeting of MDSCs using rapamycin may emerge as a promising strategy for the prevention and treatment of HF. However, the specific mechanisms underlying rapamycin-induced MDSC accumulation remain unclear. Our study aimed to clarify the effects of rapamycin on the recruitment and function of MDSCs in HF, exploring new therapeutic options for the prevention and treatment of HF. METHODS: We used transverse aortic constriction surgery and isoproterenol injection to establish HF models. Flow cytometry, reverse transcription polymerase chain reaction, transcriptomics and western blot were used to explore the regulation of rapamycin on recruitment and function of MDSCs in HF. Furthermore, rapamycin and granulocyte-macrophage colony-stimulating factor (GM-CSF) were combined to induce exogenous MDSCs from bone marrow cells. RESULTS: Rapamycin promotes the recruitment of MDSCs by inhibiting their maturation and differentiation via suppression of the Wnt signaling in HF mice and enhanced the immunosuppressive function of MDSCs via the NF-κB signaling. Furthermore, exogenous MDSCs induced by rapamycin and GM-CSF can significantly alleviate transverse aortic constriction-induced cardiac dysfunction. CONCLUSIONS: The pharmacological targeting of MDSCs using rapamycin is a promising strategy for the prevention and treatment of HF.

Assuntos

RESUMO

Whole-exome sequencing (WES) is frequently utilized in diagnosing reproductive genetic disorders to identify various genetic variants. Canonical ±1,2 splice sites are typically considered highly pathogenic, while variants at the 5' or 3' ends of exon boundaries are often considered synonymous or missense variants, with their potential impact on abnormal gene splicing frequently overlooked. In this study, we identified five variants located at the last two bases of the exons and two canonical splicing variants in five distinct families affected by reproductive genetic disorders through WES. Minigene analysis, RT-PCR and Quantitative Real-time PCR (RT-qPCR) confirmed that all seven variants induced aberrant splicing, with six variants altering gene transcriptional expression levels. These findings underscore the crucial role of splice variants, particularly non-canonical splice sites variants, in reproductive genetic disorders, with all identified variants classified as pathogenic.