CTRL Group Research Articles

Immune Checkpoint Blockade Combined with AbnobaViscum® Therapy is Linked to Improved Survival in Advanced or Metastatic Non-Small-Cell Lung Cancer Patients: A Registry Study in Accordance with the ESMO Guidance for Reporting Real-World Evidence

Background: Recent advancements in cancer treatment have shown the potential of immune checkpoint blockade (ICB) plus Viscum album L. therapy in improving survival rates for patients with advanced or metastatic non-small-cell lung cancer (NSCLC). The objective of this study was to investigate factors associated with improved survival in NSCLC patients treated with a combination of ICB and abnobaViscum®. Methods: Patients with advanced or metastatic NSCLC from the accredited Network Oncology registry were included in this real-world data study adhering to ESMO-GROW criteria with ethics approval. Survival outcomes were compared between patients receiving ICB therapy alone versus those receiving combinational ICB plus abnobaViscum® therapy using Kaplan–Meier and multivariable Cox proportional hazard analysis. Results: Among 300 patients (median age 68 years; male/female ratio 1.19), 222 received ICB alone (CTRL group) and 78 received combinational therapy (COMB group). Overall survival was significantly prolonged in the COMB group by 7 months compared to CTRL (13.8 months vs. 6.8 months, p = 0.005) with a survival rate of 16.5% in the COMB group vs. 8.0% in the CTRL group. In programmed death-ligand 1 positive (≥1%) patients treated with first-line ICB, the addition of abnobaViscum® reduced the adjusted hazard of death by 75% (aHR: 0.25; 95%CI: 0.11–0.60, p = 0.02). Conclusions: The addition of abnobaViscum® to ICB is significantly associated with improved survival in patients with advanced or metastatic NSCLC patients, irrespective of age, stage, Eastern cooperative oncology group status, surgery, or radiation. Potential mechanisms include immune modulation, reduced primary ICB resistance, and tumor microenvironment modifications. The findings warrant further validation in randomized controlled trials or registry-based randomized controlled trials. Trial registration: The study was registered (DRKS00013335).

Cryptosporidium parvum infection alters the intestinal mucosa transcriptome in neonatal calves: impacts on epithelial barriers and transcellular transport systems

IntroductionCryptosporidium parvum (C. parvum) is the most prevalent enteric protozoan parasite causing infectious diarrhea in neonatal calves worldwide with a direct negative impact on their health and welfare. This study utilized next-generation sequencing (NGS) to deepen our understanding of intestinal epithelial barriers and transport mechanisms in the pathophysiology of infectious diarrhea in neonatal calves, which could potentially unveil novel solutions for treatment.MethodsAt day 1 of life, male Holstein-Friesian calves were either orally infected (n = 5) or not (control group, n = 5) with C. parvum oocysts (in-house strain LE-01-Cp-15). On day 8 after infection, calves were slaughtered and jejunum mucosa samples were taken. The RNA was extracted from collected samples and subjected to sequencing. Differentially expressed genes (DEG) between the infected and CTRL groups were assessed using DESeq2 at a false discovery rate < 0.05 and used for gene ontology (GO) and pathway enrichment analysis in Cytoscape (v3.9.1).Results and discussionTo study the pathophysiology of infectious diarrhea on intestinal permeability, 459 genes related to epithelial cell barrier integrity and paracellular and transmembrane transport systems were selected from 12,908 identified genes in mucus. Among, there were 61 increased and 109 decreased gene transcripts belonged to adhesion molecules (e.g. ADGRD1 and VCAM1), ATP-binding cassette (ABC, e.g. ABCC2 and ABCD1) and solute carrier (SLC, e.g. SLC28A2 and SLC38A3) transporters, and ion channels (e.g. KCNJ15). Our results suggest deregulation of cellular junctions and thus a possibly increased intestinal permeability, whereas deregulation of ABC and SLC transporters and ion channels may influence the absorption/secretion of amino acids, carbohydrates, fats, and organic compounds, as well as acid-based balance and osmotic hemostasis. Besides pathogen-induced gene expression alterations, part of the DEG may have been triggered or consequently affected by inflammatory mechanisms. The study provided a deeper understanding of the pathophysiology of infectious diarrhea in neonatal calves and the host-pathogen interactions at the transcript level. For further studies with a particular focus on the transport system, these results could lead to a new approach to elucidating pathophysiological regulatory mechanisms.

Gr-1 blockade remodels the immunosuppressive microenvironment induced by incomplete microwave ablation of hepatocellular carcinoma

BackgroundAblation is one of the main methods for local treatment of hepatocellular carcinoma (HCC). Different from radiofrequency ablation (RFA), microwave ablation (MWA) is not limited by tissue conductivity, and can use multiple electrodes at the same time to improve ablation efficiency. In addition, MWA can form a larger ablation area, which makes it possible to completely ablate large HCC. However, MWA may be incomplete due to factors such as larger tumors or tumors in high-risk areas. The mechanism by which the cellular and tumor immune microenvironment (TIME) is involved in the in vitro effects of incomplete microwave ablation (iMWA) needs to be further elucidated.MethodsH22 tumor-bearing C57BL/6 mice were treated with iMWA with several combinations of ablation power and time duration. The effects of iMWA on the genes of HCC cancer cells and the TIME were investigated by RNA sequencing, mass cytometry, immunohistochemistry, and immunofluorescence. The effect of iMWA in combination with anti-Gr-1 on HCC tumor growth was also evaluated.ResultsThermal stress generated by iMWA induced coagulative necrosis and apoptosis in the region of the ablation center of HCC. RNA sequencing analysis showed that iMWA can boost chemokine CXCL5, which was further confirmed by quantitative real time polymerase chain reaction (qRT-PCR). Mass cytometry results showed that relative to Ctrl group, iMWA-treated led to decreased CD4+ T, CD8+ T, Natural killer (NK), macrophages including both M1 and M2 types but increased monocytes and bone marrow-derived suppressor cells (MDSC). Therefore, inhibiting MDSC is the main target in the later stage of iMWA. In vivo results showed that the tumor volume and weight of iMWA+ anti-Gr-1 group were significantly reduced compared with iMWA+ anti-IgG group. In addition, the merged expressions of CD11b and Gr-1 proteins were found reduced in the iMWA+ anti-Gr-1 group compared with the iMWA+ anti-IgG group by immunofluorescence staining. Immunohistochemistry suggested that CD8 was enriched in the iMWA+ anti-Gr-1 group but not in the iMWA+ anti-IgG group.ConclusionOur data suggests that iMWA and Gr-1 blocking combined therapy can further inhibit HCC growth and significantly improve the CD8+ T cells in the mouse subcutaneous tumor model, which brings good news to HCC patients.

Therapeutic Efficacy of Persimmon Leaf Flavonoids Extract in Ischemic Cerebrovascular Disease

In this research, the effects of persimmon leaf extract in treating ischemic cerebrovascular disease (ICVD) were analyzed, and the therapeutic outcomes were evaluated using techniques such as computed tomography (CT) angiography. Firstly, the impact of extraction conditions on the yield of flavonoids from persimmon leaves was assessed, and their scavenging rates (SRs) against three types of free radicals (DPPH, hydroxyl, and superoxide anion) were measured. Subsequently, 80 patients with ICVD were enrolled. Before treatment, patients’ carotid artery atherosclerotic plaque composition was evaluated using gemstone spectral CT, and the effects of conventional therapy (Ctrl group) were compared with those of treatment using persimmon leaf extract (Test group) on the patients’ blood lipids (BLs), blood glucose (BG) levels, hemorheology, hemodynamics, and CT angiography/perfusion imaging characteristics. Additionally, the post-treatment recurrence rate was recorded. The results revealed that the solvent concentration, ultrasonic power, temperature, time, and solid-liquid ratio markedly influenced the yield of persimmon leaf extract. Furthermore, the persimmon leaf extract demonstrated remarkable efficacy in scavenging three types of free radicals, namely DPPH, hydroxyl, and superoxide anion. The clinical results demonstrated that the main components of carotid artery plaque in ICVD patients were lipids, fibrous matrix, and hemorrhage, and the spectral curve combined with CT values accurately differentiated these plaque components. Patients in Test group exhibited notable improvements versus Ctrl group (P <0.05) in their serum lipid profiles (reduced total cholesterol (TC), triglyceride (TG), and high-density lipoprotein cholesterol (HDL-C) levels, increased low-density lipoprotein cholesterol (LDL-C) levels), BG levels (decreased fasting BG (FBG), glycosylated hemoglobin (HbA1c), and postprandial 2-hour BG (2h-PG) levels), hemorheological parameters (decreased low-shear whole blood viscosity (LSBV), high-shear whole blood viscosity (HSBV), blood viscosity (PV), packed cell volume (PCV), platelet adhesion rate (PAR), and fibrinogen (FIB) levels), transcranial Doppler ultrasound parameters (increased mean blood flow velocity (Vm), peak systolic velocity (PSV), and end-diastolic velocity (EDV) of middle cerebral artery (CA), anterior CA, and basilar artery), and CT angiography with perfusion imaging parameters (increased cerebral blood flow (rCBF) and region cerebral blood volume (rCBV), shortened time to peak (TTP)). Additionally, recurrence rates of transient ischemic attack and cerebral infarction were lower in Test group compared to Ctrl group. In summary, the findings suggest that persimmon leaf extract possesses potent antioxidant properties. Spectral CT can identify and distinguish carotid artery plaque components. Furthermore, persimmon leaf extract effectively improved blood lipids, blood pressure, hemorheology, hemodynamics, and cerebral perfusion status, thereby reducing the risk of recurrence in ICVD.

Therapeutic efficacy of CRISPR RNA nanoparticles on cervical cancer disease using ultrasound elastography imaging

This work aimed to evaluate the therapeutic efficacy of clustered regularly interspaced short palindromic repeats (CRISPR) ribonucleic acid (RNA) nanoparticles (NPs) in the therapy of cervical cancer (CC) using ultrasound elastography imaging. Using poly(β-amino esters) (PBAE), CRISPR RNA NPs encapsulating Human Papillomavirus 16 (HPV16) E7, namely Poly(β-amino ester) (PBAE)/CRISPR16E7-GFP and PBAE/shRNA-16E7, were prepared. The characterization of these NPs was conducted using transmission electron microscopy (TEM) and laser diffraction particle size analysis. The cytotoxicity of PBAE/CRISPR16E7-GFP and PBAE/shRNA-16E7 NPs on CC cell lines SiHa, HeLa, and CaSki was assessed using the cell counting kit-8 (CCK-8) assay. Sixty female Sprague Dawley (SD) rats were rolled into six groups randomlyas follows: the blank Ctrl group (Group A, no treatment), the model group (Group B, CC model), the pSpCas9-GFP plasmid group (Group C, CC model + pSpCas9-GFP), the PBAE/CRISPR-16E7 group (Group D, CC model + PBAE/CRISPR-16E7), the pSIREN-U6-shRNA-CMV-iRFP plasmid group (Group E, CC model + pSIREN-U6-shRNA-CMV-iRFP), and the PBAE/shRNA-16E7 group (Group F, CC model + PBAE/shRNA-16E7), each consisting of 10 rats. The impact of NPs on the expression levels (ELs) of HPV16E7 and RB1 proteins in tumor tissues was assessed using Western Blot (WB) analysis. Additionally, ultrasound elastography imaging was employed to analyze the strain ratio (SR) and shear wave velocity (SWV) in lesions of various rat groups. The results indicated that the PBAE/CRISPR16E7-GFP NPs exhibited a particle size of 125.64 ± 7.8 nm, a zeta potential of 19.17 ± 3.61 mV, and a polydispersity index (PDI) of 0.281 ± 0.03. These NPs demonstrated a regular spherical structure with a compact morphology. The PBAE/shRNA-16E7 NPs displayed a particle size of 112.93 ± 9.1 nm, a zeta potential of 13.54 ± 1.39 mV, and a PDI of 0.185 ± 0.03. They exhibited a uniform distribution of spherical shape with regularity in size. The plasmids within PBAE/CRISPR16E7-GFP and PBAE/shRNA-16E7 NPs exhibited distinct trends of burst and sustained release. The viability of SiHa, HeLa, and CaSki cells remained unaffected by PBAE/CRISPR16E7-GFP NPs. At various mass ratios, neglectable differences (P > 0.05) were observed in cell viability of SiHa, HeLa, and CaSki cells treated with PBAE/CRISPR16E7-GFP and PBAE/shRNA-16E7 NPs relative to Ctrl group. Relative to Group B, both Groups C and E exhibited a drastic decrease in tumor volume, tumor mass, and HPV16E7 protein ELs (P < 0.05), whereas Groups D and F showed a notably greater reduction in tumor volume, tumor mass, and HPV16E7 protein ELs (P < 0.01). The HPV16E7 protein ELs, SR, and SWV values greatly increased (P < 0.001) in tumors of Group B rats while the EL of RB1 protein notably decreased (P < 0.001) versus Group A. Groups C and E demonstrated elevated HPV16E7 ELs, SR, and SWV values (P < 0.05), with a marked decrease in RB1 protein EL (P < 0.001). Group F exhibited a notable reduction in HPV16E7 protein levels, SR, and SWV values (P < 0.001).The CRISPRRNA NPs composed of PBAE encapsulating HPV16 E7, namely PBAE/CRISPR16E7-GFP and PBAE/shRNA-16E7 plasmids, demonstrated a significant role in CC therapy. These NPs hold potential application value in the treatment of HPV-related CC, indicating their importance in addressing this particular disease.

A comparative study of the effects of crude chicory and inulin on gut health in weaning piglets

Dysfunction of the host-microbial balance and an impaired intestinal barrier can trigger inflammation and increase the antigen penetration. Inulin, commonly extracted from chicory root, is a prebiotic beneficial to gut health. The objective of this study was to compare the effect of chicory flour to inulin on gut health, few weeks after weaning. Two dose-dependent experiments (E1 and E2) were performed sequentially, each consisting of 80 castrated male piglets, weaned at day 21 and subsequently divided in 3 groups with ad libitum feed: control (Ctrl), inulin (INU) and crude chicory flour (CHI). For INU and CHI groups, a daily supplementation with the equated ‘inulin content’ increasing weekly was done by oral force-feeding, while the Ctrl groups received an isotonic sucrose solution. For E1, these doses were 1.5 g/day, 2 g/day and 2.5 g/day in W1, W2 and W3, respectively. For E2, these doses were 3 g/day, 4 g/day and 5 g/day in W1, W2 and W3, respectively. For each experiment at W0, W1 and W3, eight piglets per group were euthanized to assess gut structural and functional parameters. In E1, the CHI had lower average daily calorie intake (kcal/day) only at W3, while in E2 it was consistently lower than Ctrl and INU. In W3 of E2, CHI showed improved villi-to-crypt ratio and lower diarrhea occurrence than INU and Ctrl. Both supplemented groups in E2 showed higher butyrate production and lower D-xylose permeability (W3), compared to Ctrl. Interestingly, in E2, CHI had a more dominant effect on increasing the abundance of health promoting genera like Catenisphaera, Butyricicoccus, etc. and decreasing harmful genera like Erysipelotrichaceae_UCG-002 and Turicibacter. In E2, on W3 several inflammatory target genes (CXCL10, IL18, TNFα) and inflammation signalling genes (MyD88, NFκB1) were downregulated in ileum of INU and CHI. In colon, both chicory and inulin, proved to be beneficial, as the inflammation signalling and inflammatory targets genes NFκB1, DEFβ4A, TLR2 and IFNα were significantly downregulated. Therefore, crude chicory flour might also be a promising cost-effective alternative supplement to improve gut health in weaned piglets.

Transcriptome Analysis of Porcine Immune Cells Stimulated by Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) and Caesalpinia sappan Extract.

The current level of knowledge on transcriptome responses triggered by Caesalpinia sappan (CS) extract in porcine peripheral blood mononuclear cells (PBMCs) after porcine reproductive and respiratory syndrome virus (PRRSV) infection is limited. Therefore, in the present study, we aimed to detect significant genes and pathways involved in CS extract supplementation responsiveness of PBMCs after PRRSV infection. RNA sequencing was conducted on PBMCs, which were isolated from six weaned piglets. The resultant transcriptional responses were examined by mRNA sequencing. Differential expression analysis identified 263 and 274 differentially expressed genes (DEGs) between the PRRSV and CTRL groups, and the PRRSV+CS and CTRL groups, respectively. Among these, ZNF646 and KAT5 emerged as the most promising candidate genes, potentially influencing the interaction between PRRSV-infected PBMCs and CS extract supplementation through the regulation of gene networks and cellular homeostasis during stress. Two pathways were detected to be associated with CS extract supplementation responsiveness: the cellular response to stress pathway and the NF-kB signaling pathway. Consequently, our study reveals a novel mechanism underlying cellular stress response and the NF-κB signaling pathway in PRRSV-infected PBMCs, and identifies a potential application of CS extract for activating the NF-κB signaling pathway. In conclusion, by supplementing CS extract in PBMC cells infected with PRRSV, we found that CS extract modulates PRRSV infection by inducing cellular stress, which is regulated by the NF-κB signaling pathway. This induced stress creates an adverse environment for PRRSV survival. This study contributes to a deeper understanding of the therapeutic targets and pathogenesis of PRRSV infection. Importantly, our results demonstrate that CS extract has the potential to be a candidate for modulating PRRSV infection.

Meta-analysis of the efficacy and impact on cardiac function of sodium-glucose cotransporter 2 inhibitor Empagliflozin in heart failure patients.

Currently, there is no comprehensive systematic review available to comprehensively assess the efficacy and safety of Empagliflozin and other sodium-glucose cotransporter 2 inhibitors in the treatment of heart failure (HF). This study employed a meta-analysis approach to systematically evaluate the therapeutic effects of Empagliflozin in HF patients and its impact on cardiac function. The keywords including "heart failure," "HF," "cardiac failure," "cardiac disease," "Empagliflozin," and "sodium-glucose cotransporter 2 inhibitors" were utilized to search for relevant clinical studies on Empagliflozin in the treatment of HF in various databases, such as China National Knowledge Infrastructure, Wanfang, VIP Chinese Medical Journal Database, PubMed, MEDLINE, Embase, Cochrane Library, Springer, and Science Direct. The studies included patients with HF who received drug treatment. Data on baseline characteristics and posttreatment outcomes, including HF hospitalization (HHF), cardiovascular mortality, all-cause mortality, estimated glomerular filtration rate changes, Kansas City Cardiomyopathy Questionnaire quality of life (QoL) scores, N-terminal pro-B-type natriuretic peptide, left ventricular ejection fraction, hematocrit, and other relevant indicators were collected. Meta-analysis was conducted using RevMan5.3 to analyze the extracted data. A total of 15 studies were included in the final analysis, comprising 36,917 patients with HF. Among them, 18,486 patients were in Empagliflozin group, and 18,431 patients were in control (Ctrl) group. The results of the meta-analysis demonstrated that, relative to Ctrl group, Empagliflozin group showed a substantially lower HHF rate, a substantial improvement in estimated glomerular filtration rate changes, a reduced cardiovascular mortality rate, a higher Kansas City Cardiomyopathy Questionnaire QoL score, increased hematocrit values, reduced N-terminal pro-B-type natriuretic peptide changes, and enhanced left ventricular ejection fraction changes. These findings suggest that remarkable improvements in various outcomes compared to the Ctrl group. The sodium-glucose cotransporter 2 inhibitor Empagliflozin markedly reduces the HHF rate and cardiovascular mortality in HF patients. It also improves patients' QoL, enhances renal function, and increases cardiac function while reducing both, the preload and afterload.

Transcriptomics changes of calcitonin gene-related peptide in mitigating lipopolysaccharide-induced septic cardiomyopathy

Septic cardiomyopathy (SCM), a complication initiated by sepsis, presents a significant clinical challenge, leading to increased mortality rates. However, the mechanisms of SCM have not been fully uncovered. Our study involved analyzing RNA sequencing (RNA-seq) data from rat heart tissue, along with utilizing molecular docking and molecular dynamics (MD) simulations, to discover key targets and potential pharmacological actions of the calcitonin gene-related peptide (CGRP) against SCM. A lipopolysaccharide-induced SCM model was established in rats (LPS 10 mg/kg, intraperitoneal (i.p.)). Thereafter, the myocardial tissues from the three groups of rats (Ctrl group, LPS group, and CGRP group) (n = 5) were extracted and underwent RNA-seq, followed by bioinformatics analyses. The qPCR-validated hub targets potentially interacting with CGRP were identified. Following this, homology modeling was utilized to obtain the 3D structure of hub targets, and molecular docking was conducted to evaluate the interaction between CGRP and hub targets. MD simulations (300 ns) were performed to confirm the findings further. Our findings demonstrated that CGRP significantly lowered mortality in SCM rats. 633 DEGs were affected by LPS, contrasted with the Ctrl group. 96 DEGs were affected by CGRP compared to the LPS group. In total, ten fully annotated CGRP-triggered hub genes were obtained. The molecular docking and MD simulations indicate that the relationship between CGRP and eight hub genes is extremely strong. This research offers a thorough examination of the possible objectives and fundamental molecular processes of CGRP in combating SCM, laying the groundwork for investigating the potential protective mechanisms of CGRP against SCM.

Effect of Corneal Cross-linking on Biomechanical Properties of Swollen Rabbit Corneas

Corneal cross-linking (CXL) is an effective method to prevent the progression of keratoconus. CXL combined with hypotonic riboflavin solution is a modified treatment for thin corneas, which are deemed to be below the safe thickness threshold. In this study, rabbit corneas were subjected to different hydration levels using different osmolarity of riboflavin dextran solutions before CXL. Inflation testing was performed to evaluate the corneal biomechanical stiffening effect of hypotonic riboflavin solutions crosslinking. One-month post-CXL, the stromal demarcation line depth (DLD) and the biomechanical property parameter – tangent modulus (Et) – were measured. All CXL groups showed higher Et than the corresponding Ctrl groups (all P < 0.001), however, the Et values showed no statistical differences between the CXL-ed groups with different hydration levels (all P > 0.05). The relative depth ratio of DLD to total corneal thickness (TCT) did not show significant differences (P > 0.05), while the DLD was statistically different in three CXL groups (P < 0.001). The research suggested that riboflavin solutions with different osmolarities are suitable for preoperative swelling of corneas with different thickness ranges. Furthermore, crosslinking with hypotonic riboflavin solutions has no significant effect on corneal biomechanical improvement under a certain degree of hydration.

Intact DNA repair in differentiated cardiomyocytes is essential for maintaining cardiac function in response to pressure overload in mice

Abstract Introduction DNA in every cell is continuously damaged and DNA repair mechanisms are essential for protection against DNA damage-induced aging-related diseases. For example, deficient repair of endogenously generated DNA damage in mice with cardiomyocyte-restricted inactivation of Xpg, is associated with progressive heart failure. Purpose Here we tested the hypothesis that unrepaired DNA damage in differentiated cardiomyocytes increases cardiac vulnerability in response to hemodynamic overload. Methods To increase the burden of spontaneous DNA damage, we generated mice with cardiomyocyte-restricted inactivation of DNA repair endonuclease XPG. At 8 weeks of age, αMHC-Xpgc/- and control (Ctrl) mice were subjected to pressure overload by transverse aortic constriction (TAC). Eight weeks after TAC, left ventricular (LV) function was assessed using echocardiography and hemodynamic measurements, followed by molecular and histological analyses. Results Cardiomyocyte-restricted inactivation of Xpg resulted in systolic as well as diastolic LV dysfunction (Table). TAC-induced LV hypertrophy is similar in both groups (Ctrl 38%; αMHC-Xpgc/- 34%). In Ctrl mice, LV hypertrophy was accompanied by minimal LV dilation and modest changes in systolic and diastolic LV function. Conversely, TAC in αMHC-Xpgc/- produced severe LV dysfunction and resulted in overt congestive heart failure, demonstrated by aggravation of LV dilation, and marked increases in LV end-diastolic pressure, left atrial weight and lung fluid weight, which was accompanied by further increases in the expression levels of the hypertrophic marker genes atrial natriuretic peptide and beta-myosin heavy chain (Table). Moreover, lectin staining revealed a decrease in capillary density and TUNEL staining revealed further elevated levels of myocardial apoptosis. In addition, a significant increase of myocardial collagen content was observed. Conclusion Cardiomyocyte-restricted loss of DNA repair protein XPG increases cardiac vulnerability to develop heart failure in response to pressure overload. These findings underscore the importance of genomic stability for maintenance of cardiac function, not only during basal conditions, but also in response to a pathological stimulus.

Efficacy of atorvastatin on renal function in patients with contrast-induced nephropathy after percutaneous coronary intervention

BackgroundAt present, the clinical methods for preventing and treating contrast-induced nephropathy (CIN) are limited, and statins can play a better role during this process. So, we aimed to assess the atorvastatin on renal function in nephropathy patients after percutaneous coronary intervention (PCI).MethodsIn this work, 100 elderly patients with coronary heart disease (CHD) were selected into an experimental group (Exp group, 50 cases, 40 mg/d po atorvastatin) and a control group (Ctrl group, 50 cases, 10 mg/d po atorvastatin). The renal function indicators, blood routine indicators, and the incidence of adverse reactions (ARs) were compared between patients in Exp and Ctrl groups.ResultsAfter surgery, the levels of serum creatinine (SCr), blood urea nitrogen (BUN), cystatin C (CysC), high-sensitivity C-reactive protein (hs-CRP), and interleukin (IL6) in patients in the Exp group were much lower, and the levels of estimated glomerular filtration rate (eGFR) and superoxide dismutase (SOD) were higher (all P < 0.05). Meanwhile, the incidences of ARs during hospitalization between patients in the Exp and Ctrl groups were all 8%, showing no observable difference (P > 0.05). Compared with conventional doses of atorvastatin, high-dose atorvastatin can effectively prevent renal function damage in patients with CIN, decrease the inflammation and oxidative stress in patients, and will not increase the risk of ARs during hospitalization.ConclusionTaken together, high-dose atorvastatin can be applied in treating patients with CHD after PCI due to its excellent efficacy and high safety.

Empagliflozin protects the heart from atrial fibrillation in rats through inhibiting the NF-κB/HIF-1α regulatory axis and atrial remodeling

Atrial fibrillation (AF) is the most common form of sustained cardiac arrhythmia. The current study aimed to investigate the potential of empagliflozin (EMPA) to protect against acetylcholine (ACh)/calcium chloride (CaCl2)-induced AF in rats and elucidate the possible underlying mechanism of action. Rats were randomly assigned to five groups, as follows: CTRL group: received 1 ml/kg isotonic saline; AF group: received 1 ml/kg induction mixture of ACh/CaCl2 (60 µg ACh and 10 mg CaCl2 per ml); EMPA group: received 30 mg/kg EMPA; AF + EMPA10 group: received the induction mixture concurrent with 10 mg/kg EMPA; AF + EMPA30 group: received the induction mixture concurrent with 30 mg/kg EMPA. Our results showed that EMPA administration inhibited the AF-related electrocardiographic abnormalities and decreased the serum brain natriuretic peptide levels. EMPA treatment maintained the cardiac redox balance, as indicated by reduced levels of the lipid peroxidation biomarker malonaldehyde while enhancing the antioxidant glutathione levels. Moreover, EMPA markedly repressed ACh/CaCl2-induced C-reactive protein, tumor necrosis factor, and interleukin-6 production. Interestingly, EMPA administration strongly suppressed cardiac transforming growth factor beta1, collagen type I, and alpha-smooth muscle actin expression levels in the AF rats. These results were consistent with our histopathological findings, which revealed the ameliorative effect of EMPA on AF-induced inflammatory and fibrotic lesions. Mechanistically, EMPA dose-dependently downregulated nuclear factor-kappa B (NF-κB) and hypoxia-inducible factor (HIF)-1α expressions. Besides, it attenuated the pro-apoptotic active caspase-3 while augmenting the anti-apoptotic B-cell lymphoma 2 expressions. Furthermore, EMPA dose-dependently suppressed cardiac phosphatidylinositol 3-kinase (PI3K)/Akt signaling. In conclusion, this study demonstrates that EMPA intervention, within AF induction, protects against ACh/CaCl2-induced AF in rats, exerting powerful antioxidant, anti-inflammatory, anti-fibrotic, and anti-apoptotic effects. These effects are mainly mediated through the targeting of the NF-κB/HIF-1α regulatory axis in a dose-dependent manner.

8124 Prevalence and Characteristics of Patients with Hypothyroidism in the Danish Blood Donor Study

Abstract Disclosure: N.L. Frisk: None. J.R. Shorter: None. O.B. Pedersen: None. L.T. Dalgaard: None. Introduction: While impaired thyroid function leading to subclinical hypothyroidism and overt hypothyroidism is common in Denmark, the knowledge about the paths by which patients’ progress to impaired thyroid function is limited. Undiagnosed hypothyroidism as well as subclinical hypothyroidism constitute serious risk factors for development of cardio metabolic disease. The Danish Blood Donor Study (DBDS) is a large longitudinal cohort and biobank containing information and samples dating more than 10 years back. Blood donors donate blood every 3-6 months and each time provide a sample for research. Research question: The aim of the study was to obtain up-to-date information about prevalence and characteristics of patients with hypothyroidism in a Danish longitudinal cohort of blood donors. Methodology: The study employed a cross-sectional design. Hypothyroidism was defined based on at least two prescriptions of levothyroxine (LT4) and used for calculations of prevalence. Case numbers were compared to all controls that were currently not being prescribed LT4 nor anti-thyroid drugs (carbimazole, propylthiouracil, thiamazole). We compared cases of hypothyroidism to two different control (ctrl) groups: 1) All the participants of the DBDS having TSH levels in the normal range, and 2) Normothyroid participants matched to hypothyroid cases based on age and sex. Incidence was determined based on DBDS participants developing hypothyroidism after being enrolled into the DBDS during a 10-year follow-up. Men and women were compared separately among cases and controls. Students’ t-test was used to compare cases and controls, and corrected using Bonferroni. Major results: There were 3397 cases of hypothyroidism and 153791 ctrl subjects in the DBDS, giving a prevalence of 2.2%. 85% of cases were female, while among ctrl 50% were female. Average age of cases was 52±13 years (mean±SD) for women and 58±13 for men, while ctrls were 45±13 and 48±14 years, respectively (both P&amp;lt;0.001). BMI among cases were: 25.8±4.7 for women and 26.8±3.9 kg/m2, while cases were 25.1±4.4 and 26.0±3.6, respectively (both P&amp;lt;0.001). TSH levels were, for female cases: 4.6±4.0 iu/mL and 5.7±4.9 for men, while ctrls were 1.8±1.1 and 1.7±1.1, respectively (both P&amp;lt;0.001). Free (F) T4 levels were, for female cases: 14.4±6.4pmol/L and male: 14.0±4.6, while ctrls were: 14.8±4.5 and 15.0±3.9 respectively (P&amp;lt;0.003 for females, P&amp;lt;0.0001 for males), while FT3 levels were for cases: 4.3±1.7 and 4.3±1.4 pmol/L and 4.5±2.0 and 4.7±1.5 for ctrls (both P&amp;lt;0.0001). Conclusion: Hypothyroidism is less prevalent among blood donors than reported in the general Danish population, which is accordance with ‘the healthy donor’ bias. Hypothyroid donors were on average older and with higher BMI than normothyroid controls. Donors with hypothyroidism were generally well regulated, but despite that FT4 and FT3 levels were lower than ctrls. Presentation: 6/1/2024

Morroniside repairs atrazine-induced skin damage by ameliorating lipid metabolism disorders and inhibiting ferroptosis

Morroniside (MOR) has shown great potential in treating atrazine (ATZ)-induced skin damage. This study aims to elucidate MOR's mechanism of action in mitigating lipid metabolism disorders and inhibiting ferroptosis to repair ATZ-induced skin damage. Twenty C57BL/6 mice were divided into four groups: the control group, the ATZ group, the MOR-H group and the MOR-L group, each comprising five mice. Following a one-month intervention, mouse skin tissues were harvested for untargeted lipid metabolomics analysis. Subsequently, the samples were assessed for indices related to ferroptosis. Untargeted lipid metabolomics analysis showed 127 differential metabolites in the ATZ vs. Ctrl group. There were 57 differential metabolites in the MOR-L vs. ATZ group. 34 differential metabolites in the MOR-H vs. ATZ group. the most obvious lipid reversal occurred after MOR-L administration, which primarily involved phospholipids, ceramides, and sphingomyelins. The levels of GPX4, Ferritin, MDA, SOD and GSH-PX, ferroptosis-related indicators, and the levels of p21 and p53, apoptosis-related indicators, were most significantly regressed in the MOR-L group (all P < 0.05). MOR may delay cellular aging and correct skin damage by reversing ATZ-induced lipid metabolism disorders, inhibiting ferroptosis and excessive oxidative stress occurrence.

When the neighbors are noisy: effect of social challenge in collateral pens of stressed animals.

Regrouping practices are frequent in pig production, altering hierarchy and triggering aggressive behaviors. The present study aimed to investigate the physiological responses of piglets to an experimental model designed to induce stress through systematic social mixing in two trials. In Trial A, a total of 144 crossbred piglets (25 days postweaning) housed in one room within 36 pens (four piglets/pen) were used and randomly assigned to either a control group (piglets maintained in their pen, Ctrl-A) or a social challenge group (piglets mixed, SC-A). In Trial B, the same number of animals (33 days postweaning) and crossbreed line was used, and each piglet was assigned either to a control group (Ctrl-B) or a social challenge group (SC-B) in two independent rooms (rooms Ctrl and SC, 12 pens/ room, six piglets/pen). The social challenge consisted of daily moves of three out of four pen mates and five out of six pen mates, for Trials A and B, respectively. In the Ctrl groups, all piglets stayed in their original pen. Before the 1st mixing day and at the end of the 3rd mixing day, saliva (cortisol concentration) and blood (cortisol concentration changes, hemogram, and immunologic activation) samples were collected from two random piglets per pen. Skin lesion scores of all piglets were also recorded on the front, middle, and rear body regions. In Trial A, the total skin lesions score was higher in the SC-A group compared to the Ctrl-A group after the social challenge (0.53 vs. 0.17; p < 0.05), but an unexpected increase between sampling days in the Ctrl-A piglets (0.06 vs. 0.17; p < 0.05) was also recorded, suggesting that Ctrl-A pigs showed similar aggressivity levels to the SC-A group. Hematological parameters hemoglobin, red blood cell counts, and leukocyte counts present similar changes in both treatment groups after the social challenge. Contrarily, in Trial B, the lesion score only increased in the piglets in room SC (0.08 vs. 0.34; p < 0.05). Results suggest that stable groups may show aggressive behaviors if they are in the same room with socially challenged pigs. Thus, the physical separation of treatment groups in social stress studies is recommended.

Loss of collagen content is localized near cartilage lesions on the day of injurious loading and intensified on day 12.

Joint injury can lead to articular cartilage damage, excessive inflammation, and post-traumatic osteoarthritis (PTOA). Collagen is an essential component for cartilage function, yet current literature has limited understanding of how biochemical and biomechanical factors contribute to collagen loss in injured cartilage. Our aim was to investigate spatially dependent changes in collagen content and collagen integrity of injured cartilage, with an explant model of early-stage PTOA. We subjected calf knee cartilage explants to combinations of injurious loading (INJ), interleukin-1α-challenge (IL) and physiological cyclic loading (CL). Using Fourier transform infrared microspectroscopy, collagen content (Amide I band) and collagen integrity (Amide II/1338 cm-1 ratio) were estimated on days 0 and 12 post-injury. We found that INJ led to lower collagen content near lesions compared to intact regions on day 0 (p < 0.001). On day 12, near-lesion collagen content was lower compared to day 0 (p < 0.05). Additionally, on day 12, INJ, IL, and INJ + IL groups exhibited lower collagen content along most of tissue depth compared to free-swelling control group (p < 0.05). CL groups showed higher collagen content along most of tissue depth compared to corresponding groups without CL (p < 0.05). Immunohistochemical analysis revealed higher MMP-1 and MMP-3 staining intensities localized within cell lacunae in INJ group compared to CTRL group on day 0. Our results suggest that INJ causes rapid loss of collagen content near lesions, which is intensified on day 12. Additionally, CL could mitigate the loss of collagen content at intact regions after 12 days.

Dual mechanism of membrane covering on GHG and NH3 mitigation during industrial-scale experiment on dairy manure composting: Inhibiting formation and blocking emissions

An industrial-scale experiment on dairy manure composting with the control group (Ctrl) and the membrane covering group (CM) was conducted to explore the effects of functional membrane covering on gas emissions, the conversion of carbon and nitrogen, and revealing the underlying mechanisms. Results indicated that CM achieved the synergistic effects on gas mitigation and improved compost product quality. CO2, CH4, N2O, and NH3 emissions were reduced by 81.8%, 87.0%, 82.6%, and 82.2%, respectively. The micro-aerobic condition formed in membrane covering compost pile together with the covering inhibiting effect dominated the mitigation effect. CM significantly downregulated the mcrA gene copies and the value of mcrA/pmoA (p < 0.01), which reduced CH4 emission. CM decreased the nirS and nirK gene copies and increased the nosZ gene copies to reduce N2O emission. Functional Annotation of Prokaryotic Taxa showed that membrane covering effectively amended part of carbon and nitrogen cycles, which stimulated the degradation of organic matter, accelerated compost maturity and reduced the gaseous emissions.

Influence of Trabecular Bone Presence on Osseodensification Instrumentation: An In Vivo Study in Sheep.

Osseodensification enhances the stability of endosteal implants. However, pre-clinical studies utilizing osseodensification instrumentation do not account for the limited presence of trabeculae seen clinically. This study aimed to evaluate the effect of osseodensification instrumentation on osteotomy healing in scenarios with and without the presence of trabecular bone. A ~10 cm incision was made over the hip of twelve sheep. Trabecular bone was surgically removed from twelve sites (one site/animal; negative control (Neg. Ctrl)) and left intact at twelve sites (one site/animal; experimental group (Exp.)). All osteotomies were created using the osseodensification drilling protocol. Each osteotomy received an endosteal implant and was evaluated after 3 or 12 weeks of healing (n = 6 animals/time). Histology revealed increased woven and lamellar bone surrounding the implants in the Exp. group relative to the Neg. Ctrl group. The Exp. group demonstrated the presence of bone fragments, which acted as nucleating sites, thereby enhancing the bone formation and remodeling processes. Bone-to-implant contact (%BIC) and bone area fractional occupancy (%BAFO) were significantly higher in the Exp. group relative to the Neg. Ctrl group both at 3 weeks (p = 0.009 and p = 0.043) and 12 weeks (p = 0.010 and p = 0.008). Osseodensification instrumentation in the presence of trabecular bone significantly improved osseointegration. However, no negative influences such as necrosis, inflammation, microfractures, or dehiscence were observed in the absence/limited presence of trabeculae.

PSLBII-3 Influence of iron supplementation on insulin and glucose dynamics in horses

Abstract Iron is an essential micromineral involved in various physiological functions such as oxygen transport. The National Research Council’s Nutrient Requirements of Horses has iron requirements set at 400 mg iron daily for a 500 kg horse at maintenance; however, a survey found that Thoroughbreds consumed well over the daily requirement at 3,900 mg of iron from hay and grain alone. Additionally, a previous study has found a correlation between hyperinsulinemia and increased ferritin, an indicator of iron body stores. Therefore, the objective of the present study was to determine the effects of iron supplementation on body iron stores as well as insulin and glucose dynamics. It was hypothesized that iron supplementation would influence body iron stores and insulin and glucose responses. Mixed-breed geldings [n = 12; 558.9 ± 74.4 kg body weight (BW)] were housed individually for the study. Horses were fed 2% of their BW in grass hay and an iron-free vitamin mineral supplement, consuming an average of 600 mg of iron daily for the first 28 d (Hay Phase). Horses were then assigned to continue on the hay diet (CTRL; n = 4) or an iron-supplemented diet (IRON; n = 8), in which an oral iron supplement was given daily in the form of ferrous sulfate, with IRON horses consuming an average of 4,000 mg of iron daily for 28 d (Supplement Phase). Oral sugar tests (OSTs) were performed at the beginning of the study as well as the end of each phase, using a 0.45 mL/kg of BW Karo Light Syrup dose to determine insulin response, with jugular venous samples taken prior to the dose and 60 min after. All statistical analysis was performed in GraphPadPrism Version 10.2.0 (GraphPad Software, Boston, MA), using analysis of variance for repeated measures and correlation analysis. All OSTs throughout the study had a significant time effect (P &amp;lt; 0.05) for both insulin and glucose. There was a time X treatment trend (P = 0.1) for insulin during the Supplement Phase and when using multiple comparisons, insulin response was significantly greater (P &amp;lt; 0.0007) at 0 versus 60 min in IRON horses. Although there was no correlation found between 60-min insulin and ferritin concentrations in both IRON and CTRL horses, mean ± SD insulin and ferritin in the IRON group were 23.6 ± 10 uU/mL and 570 ± 195.8 ng/mL, respectively, while in the CTRL group these were 12.7 ± 2.9 uU/mL and 453 ± 31.1 ng/mL, respectively. In conclusion, iron supplementation with an inorganic form at 10 times the daily requirement appears to result in increased insulin responses as well as body iron stores. Further research needs to determine if feeds naturally enriched in iron impact glucose metabolism.