Vericiguat combined with "new quadruple" therapy enhances cardiac function and life quality in patients with heart failure: a single-center prospective study.

Objective: To investigate the therapeutic effect of Vericiguat combined with "new quadruple" drugs on patients with heart failure (HF). Methods: From December 1, 2022 to February 1, 2024, 103 patients with heart failure were consecutively enrolled from the cardiology clinic or ward of the First Affiliated Hospital of Nanjing Medical University. Before enrollment, the patients' left ventricular ejection fraction (LVEF), left ventricular end diastolic diameter (LVEDD), N-terminal pro-B-type natriuretic peptide (NT-proBNP), liver and kidney function electrolytes, and Minnesota Living with Heart Failure Questionnaire (MLHFQ) and other indicators were measured. Patients diagnosed with reduced ejection fraction (HFrEF) and with heart failure with mildly reduced ejection fraction (HFmrEF) were treated with Vericiguat combined with "ARNI, BB, MRA, SGLT2i" therapy. Patients diagnosed with preserved ejection fraction (HFpEF) were treated with Vericiguat combined with "ARNI, BB, SGLT2i" therapy. The above indicators were rechecked after 1 month of treatment. Results: For all patients, comparison after treatment: LVEF (38.1 ± 8.5% vs. 43.1 ± 8.5%, P < 0.01), LVEDD (60.5 ± 8.1 vs. 58.2 ± 7.3 mm, P < 0.01), NT-proBNP (4,567.8 ± 5,163.9 vs. 1,895.6 ± 2,702.1 ng/L, P < 0.01), MLHFQ (45.72 ± 11.09 vs. 32.29 ± 9.41, P < 0.01). Further subgroup analysis showed that Vericiguat combined with "ARNI, BB, SGLT2i or MRA" improved the LVEF and reduced NT-proBNP levels in patients with HFrEF, HFmrEF or HFpEF. and improved patients' quality of life scores. The intergroup comparison showed the therapeutic effect of the combination was equivalent in HF caused by myocardial Infarction (MI), dilated cardiomyopathy (DCM) or Valvular Heart Disease (VHD). Conclusion: Vericiguat combined with the "new quadruple" therapy has a significant therapeutic effect on patients with heart failure caused by MI, DCM or VHD.

[Effect of electroacupuncture at "Fenglong" (ST 40) on liver cholesterol metabolism in hyperlipidemia rats based on AMPK/mTOR pathway].

OBJECTIVE: To observe the effects of electroacupuncture (EA) at "Fenglong" (ST 40) on the expression of adenosine 5'-monophosphate-activated protein kinase (AMPK), mammalian target of rapamycin (mTOR), and the expression of the downstream molecules related to cholesterol metabolism i.e. sterol regulatory element binding protein-2 (SREBP-2), recombinant 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGCR), and adenosine triphosphate binding cassette transporter G5/G8(ABCG5/G8) in the rats with hyperlipidemia (HPL) so as to explore the possible mechanism of EA in the intervention of HPL. METHODS: Thirty SPF-grade male SD rats were randomly divided into a blank group, a model group, an AMPK agonist group, an EA group, and an EA+AMPK inhibitor group, 6 rats in each group. The high-fat feeding method was adopted to establish HPL model. After successfully modeled, the rats in the EA group received EA intervention at bilateral "Fenglong" (ST 40), with disperse-dense wave, in the frequency of 2 Hz/100 Hz, the intensity of 1 mA. EA was given once daily, for 30 min in one intervention. In the AMPK agonist group, the intraperitoneal injection with AMPK agonist A-769662 was administered, 30 mg/kg, twice a day. In the EA+AMPK inhibitor group, the intraperitoneal injection of AMPK inhibitor Compound C was administered, 25 mg/kg, once a day, 30 min before EA intervention. In the intervention groups, the interventions were delivered continuously for 5 days a week and lasted 4 weeks. Using automated biochemical analyzer, the blood lipid-related indexes (serum total cholesterol [TC], triglycerides [TG], low-density lipoprotein cholesterol [LDL-C] and high-density lipoprotein cholesterol [HDL-C] as well as alanine aminotransferase [ALT] and aspartate aminotransferase [AST]) were detected in the rats. HE staining and oil red O staining were used to observe the morphology of liver tissue. Liver index was calculated by the weight. Using ELISA, the contents of TC and TG of liver tissue and the contents of of TC and bile acid in feces were detected. The protein phosphorylation levels of AMPK and mTOR in the liver tissue were detected using Western blot; and the positive expression of SREBP-2, HMGCR and ACBG5/G8 was detected using immunohistochemical staining. RESULTS: After modeling, the levels of serum TC, TG and LDL-C of rats in the model group, the AMPK agonist group, the EA group and the EA+AMPK inhibitor group were all higher than those in the blank group (P<0.01); and there was no statistically difference in the levels of serum HDL-C among groups (P>0.05). After intervention, compared with the blank group, in the model group, the levels of serum TC, TG, LDL-C, ALT and AST, the liver index, the levels of TC and TG in liver tissue, the levels of TC and the bile acid in feces were increased (P<0.01); HE and oil red O staining showed that the hepatocytes were disordered, and there were macrovesicular lipid droplets in the cells and the obvious lipid accumulation; the protein expression of phosphorylated AMPK (p-AMPK) in liver tissue and the ratio of p-AMPK and AMPK were reduced (P<0.01), the protein expression of phosphorylated mTOR (p-mTOR) and the ratio of p-mTOR and mTOR were elevated (P<0.01); and the positive expression of SREBP-2, HMGCR, ABCG5 and ABCG8 in liver tissue was increased (P<0.01, P<0.05). Compared with the model group, in the AMPK agonist group and the EA group, the levels of serum TC, TG, LDL-C, ALT and AST, liver indexes, the levels of TC and TG in liver tissue were reduced (P<0.01), while the levels of TC and bile acid in feces were increased (P<0.05, P<0.01); HE staining and oil red O staining showed that the hepatocytes were in order, and lipid accumulation; the protein expression of p-AMPK and the ratio of p-AMPK and AMPK in liver tissue increased (P<0.01), while the protein expression of p-mTOR and the ratio of p-mTOR and mTOR decreased (P<0.01); the positive expression of SREBP-2 and HMGCR in liver tissue was reduced (P<0.01), while that of ABCG5 and ABCG8 up-regulated (P<0.05, P<0.01) . Compared with the EA group, in the EA+AMPK inhibitor group, the levels of serum TC, TG, LDL-C, ALT and AST, liver index, the levels of TC and TG in liver tissue were increased (P<0.05, P<0.01), while the levels of TC and bile acid in feces were reduced (P<0.01); lipid accumulation was aggravated; the protein expression of p-AMPK and the ratio of p-AMPK and AMPK in liver tissue were reduced (P<0.01, P<0.05), while the protein expression of p-mTOR and the ratio of p-mTOR and mTOR elevated (P<0.05, P<0.01); the positive expression of SREBP-2 and HMGCR in liver tissue was increased (P<0.01), while that of ABCG5 and ABCG8 was down-regulated (P<0.01). CONCLUSION: EA at "Fenglong" (ST 40) can attenuate hyperlipidemia in HPL rats. It may be achieved by regulating the AMPK/mTOR pathway, inhibiting the expression of cholesterol synthesis related molecules, SREBP-2 and HMGCR, and up-regulating the expression of cholesterol excretion molecules, ABCG5 and ABCG8, thereby reducing liver cholesterol accumulation and increasing cholesterol excretion.

Social network analysis of regional transport carbon emissions in China: Based on motif analysis and exponential random graph model.

Given the significant impact of transportation-related carbon emissions on air quality and climate change, understanding the regional dynamics of these emissions is crucial. Despite numerous studies on carbon emissions, there is a lack of comprehensive analysis of China's interprovincial transport carbon emission correlation network. Based on China's provincial data from 2007 to 2021, we analyzed the network's basic structural characteristics and categorized it into four significant plates to investigate their interactions. Subsequently, motif analysis is employed to examine the micro-correlation patterns within the network, and the Exponential random graph model (ERGM) is utilized to analyze the network's formation mechanism. Findings reveal that: (1) Provinces with high correlation intensity are predominantly concentrated in the eastern region, such as Shanghai and Beijing. Additionally, provinces in the eastern region assume a central role in the transport carbon emission correlation network, mainly receiving carbon emissions from other provinces. In contrast, the western region primarily emits carbon emissions to other provinces, continuously converging towards the center. (2) The network is segmented into net beneficiary plate, net overflow plate, bidirectional spillover plate, and broker plate, with distinct roles and influences across different years. (3) Bidirectional correlation motif structures emerge as primary influencers within the network, although specific structures impede interregional communication and collaborative emission reduction. (4) Internal network's structural variables, such as mutuality, cyclic triple, and geometrically weighted edgewise shared partner, along with influencing factors including government intervention, urbanization rate, openness, fiscal expenditure on transport, and province adjacency significantly impact the formation of the transport carbon emission correlation network. The above transportation network research provides a theoretical basis for the country to promote low-carbon transportation and improve air quality, and also has important guiding significance for the cross-regional collaborative emission reduction work of provinces.

Computationally-Assisted Discovery and Assignment of a New Class of 6/6/5/5 Fused-Ring Diterpene Acting as Pregnane X Receptor Ligands from Isodon serra.

We report here the orchestration of molecular ion networking (MoIN) and a set of computationally assisted structural elucidation approaches in the discovery and assignment of a new class of rearranged 4,5-seco-abietane diterpenoids including serra A (1), which possesses an unusual 6/6/5/5 fused-ring skeleton system, together with two previously unreported diterpenoids serras B-C (2-3) and five known compounds were isolated from Isodon serra (I. serra). The structures were elucidated by spectroscopic analysis in conjunction with computationally assisted structure elucidation tools. In silico, serras A-C (1-3) bind well to PXR, suggesting their potential role in reducing inflammation. The results of serra A (1) with hPXR demonstrated agonist activity with an EC50 value of 15 µM. Serra A (1), graciliflorin F (4), gerardianin C (5), 11,12,15-trihydroxy-8,11,13-abietatrien-7-one (6), rabdosin D (7), and 15-hydroxysalprionin (8) exhibited promising anti-inflammatory activities in lipopolysaccharide (LPS)-induced RAW 267.4 cells, and their inhibition rates on NO production were more than 65% at 10 µM.

An Acid-Responsive Fluorescent Molecule for Erasable Anti-Counterfeiting.

A tetraphenylethylene (TPE) derivative, TPEPhDAT, modified by diaminotriazine (DAT), was prepared by successive Suzuki-Miyaura coupling and ring-closing reactions. This compound exhibits aggregation-induced emission enhancement (AIEE) properties in the DMSO/MeOH system, with a fluorescence emission intensity in the aggregated state that is 5-fold higher than that of its counterpart in a dilute solution. Moreover, the DAT structure of the molecule is a good acceptor of protons; thus, the TPEPhDAT molecule exhibits acid-responsive fluorescence. TPEPhDAT was protonated by trifluoroacetic acid (TFA), leading to fluorescence quenching, which was reversibly restored by treatment with ammonia (on-off switch). Time-dependent density functional theory (TDDFT) computational studies have shown that protonation enhances the electron-withdrawing capacity of the triazine nucleus and reduces the bandgap. The protonated TPEPhDAT conformation became more distorted, and the fluorescence lifetime was attenuated, which may have produced a twisted intramolecular charge transfer (TICT) effect, leading to fluorescence redshift and quenching. MeOH can easily remove the protonated TPEPhDAT, and this acid-induced discoloration and erasable property can be applied in anti-counterfeiting.

Quantifying Forms and Functions of Enterohepatic Bile Acid Pools in Mice.

BACKGROUNDS & AIMS: Bile acids (BAs) are core gastrointestinal metabolites with dual functions in lipid absorption and cell signaling. BAs circulate between the liver and distal small intestine (i.e., ileum), yet the dynamics through which complex BA pools are absorbed in the ileum and interact with host intestinal cells in vivo remain poorly understood. Because ileal absorption is rate-limiting in determining which BAs in the intestinal lumen gain access to host intestinal cells and receptors, and at what concentrations, we hypothesized that defining the rates and routes of ileal BA absorption in vivo would yield novel insights into the physiological forms and functions of mouse enterohepatic BA pools. METHODS: Using ex vivo mass spectrometry, we quantified 88 BA species and metabolites in the intestinal lumen and superior mesenteric vein of individual wild-type mice, and cage-mates lacking the ileal BA transporter, Asbt/Slc10a2. RESULTS: Using these data, we calculated that the pool of BAs circulating through ileal tissue (i.e., the ileal BA pool) in fasting C57BL/6J female mice is ∼0.3 µmol/g. Asbt-mediated transport accounted for ∼80% of this pool and amplified size. Passive permeability explained the remaining ∼20% and generated diversity. Compared with wild-type mice, the ileal BA pool in Asbt-deficient mice was ∼5-fold smaller, enriched in secondary BA species and metabolites normally found in the colon, and elicited unique transcriptional responses on addition to exvivo-cultured ileal explants. CONCLUSIONS: This study defines quantitative traits of the mouse enterohepatic BA pool and reveals how aberrant BA metabolism can impinge directly on host intestinal physiology.

Global trends in publications regarding macrophages-related diabetic foot ulcers in the last two decades.

BACKGROUND: Diabetic foot ulcers (DFUs) are one of the most severe and popular complications of diabetes. The persistent non-healing of DFUs is the leading cause of ampu-tation, which causes significant mental and financial stress to patients and their families. Macrophages are critical cells in wound healing and perform essential roles in all phases of wound healing. However, no studies have been carried out to systematically illustrate this area from a scientometric point of view. Although there have been some bibliometric studies on diabetes, reports focusing on the investigation of macrophages in DFUs are lacking. AIM: To perform a bibliometric analysis to systematically assess the current state of research on macrophage-related DFUs. METHODS: The publications of macrophage-related DFUs from January 1, 2004, to December 31, 2023, were retrieved from the Web of Science Core Collection on January 9, 2024. Four different analytical tools: VOSviewer (v1.6.19), CiteSpace (v6.2.R4), HistCite (v12.03.07), and Excel 2021 were used for the scientometric research. RESULTS: A total of 330 articles on macrophage-related DFUs were retrieved. The most published countries, institutions, journals, and authors in this field were China, Shanghai Jiao Tong University of China, Wound Repair and Regeneration, and Aristidis Veves. Through the analysis of keyword co-occurrence networks, historical direct citation networks, thematic maps, and trend topics maps, we synthesized the prevailing research hotspots and emerging trends in this field. CONCLUSION: Our bibliometric analysis provides a comprehensive overview of macrophage-related DFUs research and insights into promising upcoming research.

Red blood cell distribution width-to-monocyte ratio for predicting 90-day mortality of COVID-19 in patients with chronic kidney disease during the Omicron period: a prospective single-center study.

We aimed to test whether red blood cell distribution width (RDW) to monocyte percentage ratio (RMR) was associated with the acute-phase prognosis of coronavirus disease 2019 (COVID-19) in chronic kidney disease (CKD) patients. Prospective enrollment and 90-day follow-up of CKD patients with COVID-19 were conducted from December 1, 2022 to January 31, 2023. Demographics, clinical data, and laboratory and radiographic findings were collected, and multiple logistic regression, subgroup analysis, and receiver operating characteristic (ROC) curve analysis were performed. A total of 218 patients were enrolled, with a mean age of 59 years and 69.7% being male. The 90-day mortality rate was 24.8%. The lnRMR level was 5.18 (4.91-5.43) and emerged as an independent risk factor (OR: 3.01, 95% CI: 1.72-5.85). The lnRMR-mortality association was consistent across sex, age, CKD stage, COVID-19 vaccination, and comorbidity subgroups. The area under the ROC curve of lnRMR was 0.737 (95% CI: 0.655-0.819). Our findings indicate that lnRMR is a simple and practical predictor for identifying high-risk CKD patients during the acute phase of COVID-19.

Effect of Axial Clearance Variation on Dual-Rotor Flowmeter Performance.

This study examines the impact of axial clearance variations on the performance characteristics of a dual-rotor flowmeter (DRT-FM) through numerical simulations, with the validity of the numerical results verified by calibration experiments. The results indicate that within the range of 200 L/h to 1600 L/h, the K factors of different groups increase as clearance increases. The K factor of the 0.80 mm group is the largest, showing an average increase of around 6% compared to that of the 0.50 mm group. Additionally, Linearity E also decreased, with a minimum of 1.07% in the 0.65 mm group, significantly lower than the 3.33% in the 0.50 mm group. Furthermore, the pressure loss increased slightly, with the 0.65 mm group having the largest pressure loss; however, at a flow rate of 1600 L/h, the pressure loss only increases by 0.186 kPa compared to that of the 0.50 mm group. Flow field analysis reveals that changes in axial clearance predominantly affect pressure distribution. Larger clearances reduce low-pressure regions on upstream and downstream transition surfaces, thereby reducing energy loss due to pressure changes. Entropy analysis further demonstrates that higher axial clearance decreases energy loss in the upstream and downstream stationary domains, optimizing the DRT-FM's energy characteristics.

Microbiological profile of diabetic foot infections in China and worldwide: a 20-year systematic review.

Introduction: Pathogens causing diabetic foot infections (DFIs) vary by region globally; however, knowledge of the causative organism is essential for effective empirical treatment. We aimed to determine the incidence and antibiotic susceptibility of DFI pathogens worldwide, focusing on Asia and China. Methods: Through a comprehensive literature search, we identified published studies on organisms isolated from DFI wounds from January 2000 to December 2020. Results: Based on our inclusion criteria, we analyzed 245 studies that cumulatively reported 38,744 patients and 41,427 isolated microorganisms. DFI pathogens varied according to time and region. Over time, the incidence of Gram-positive and Gram-negative aerobic bacteria have decreased and increased, respectively. America and Asia have the highest (62.74%) and lowest (44.82%) incidence of Gram-negative bacteria, respectively. Africa has the highest incidence (26.90%) of methicillin-resistant Staphylococcus aureus. Asia has the highest incidence (49.36%) of Gram-negative aerobic bacteria with species infection rates as follows: Escherichia coli, 10.77%; Enterobacter spp., 3.95%; and Pseudomonas aeruginosa, 11.08%, with higher local rates in China and Southeast Asia. Linezolid, vancomycin, and teicoplanin were the most active agents against Gram-positive aerobes, while imipenem and cefoperazone-sulbactam were the most active agents against Gram-negative aerobes. Discussion: This systematic review showed that over 20 years, the pathogens causing DFIs varied considerably over time and region. This data may inform local clinical guidelines on empirical antibiotic therapy for DFI in China and globally. Regular large-scale epidemiological studies are necessary to identify trends in DFI pathogenic bacteria. Systematic review registration: https://www.crd.york.ac.uk/prospero/, identifier CRD42023447645.

Prediction of carbon emissions in China's construction industry using an improved grey prediction model.

As a significant source of global energy consumption and greenhouse gas emissions, the construction industry garners widespread attention due to its high carbon emissions. Anticipating its development trends is crucial for energy conservation and emission reduction. In this paper, we utilize the carbon emission data from China's national and provincial construction sectors from 2012 to 2021, employ the grey prediction model optimized by the particle swarm optimization algorithm, coupled with a metabolic algorithm, to forecast the carbon emissions of the construction industry across China and its provinces. The results demonstrate that: (1) The dynamic grey prediction model combined with the metabolism algorithm has a better prediction effect than the classical model, and the relative error is reduced from 5.103 % to 0.874 %. (2) The carbon emissions of China's construction industry will continue to rise in the next decade, but the growth rate will decrease, and the proportion of indirect carbon emissions continues to increase. (3) There is a marked regional disparity in carbon emissions, with the eastern region exhibiting higher emission levels yet slower growth. In contrast, the western region has lower emission levels but experiences faster growth. These studies provide valuable insights for both the existing approaches to energy conservation and emission reduction, as well as for future policy improvements.

Diindoles produced from commensal microbiota metabolites function as endogenous CAR/Nr1i3 ligands.

Numerous studies have demonstrated the correlation between human gut bacteria and host physiology, mediated primarily via nuclear receptors (NRs). Despite this body of work, the systematic identification and characterization of microbe-derived ligands that regulate NRs remain a considerable challenge. In this study, we discover a series of diindole molecules produced from commensal bacteria metabolites that act as specific agonists for the orphan constitutive androstane receptor (CAR). Using various biophysical analyses we show that their nanomolar affinities are comparable to those of synthetic CAR agonists, and that they can activate both rodent and human CAR orthologues, which established synthetic agonists cannot. We also find that the diindoles, diindolylmethane (DIM) and diindolylethane (DIE) selectively up-regulate bona fide CAR target genes in primary human hepatocytes and mouse liver without causing significant side effects. These findings provide new insights into the complex interplay between the gut microbiome and host physiology, as well as new tools for disease treatment.

Quantifying forms and functions of intestinal bile acid pools in mice.

Bile acids (BAs) are gastrointestinal metabolites that serve dual functions in lipid absorption and cell signaling. BAs circulate actively between the liver and distal small intestine (i.e., ileum), yet the dynamics through which complex BA pools are absorbed in the ileum and interact with intestinal cells in vivo remain ill-defined. Through multi-site sampling of nearly 100 BA species in individual wild type mice, as well as mice lacking the ileal BA transporter, Asbt/Slc10a2, we calculate the ileal BA pool in fasting C57BL/6J mice to be ~0.3 µmoles/g. Asbt-mediated transport accounts for ~80% of this pool and amplifies size, whereas passive absorption explains the remaining ~20%, and generates diversity. Accordingly, ileal BA pools in mice lacking Asbt are ~5-fold smaller than in wild type controls, enriched in secondary BA species normally found in the colon, and elicit unique transcriptional responses in cultured ileal explants. This work quantitatively defines ileal BA pools in mice and reveals how BA dysmetabolism can impinge on intestinal physiology.

Efficiency evaluation of China's transportation system considering carbon emissions: Evidence from big data analytics methods.

China's transportation sector is a vital link between production and consumption, but it also has issues with low efficiency, high carbon emissions, and technological bottlenecks. To improve efficiency and provide actionable recommendations and strategies, this study first constructs a comprehensive evaluation index system to gauge the transportation sector's inputs using panel data from different Chinese provinces from 2007 to 2021. Within the assessment system, the principal component analysis (PCA) method is used to reduce the dimension of the indexes, thereby yielding a set of adjusted inputs. Subsequently, the transportation system efficiency (TSE) is evaluated using the super-efficiency SBM-DEA model, which includes unexpected outputs such as carbon emissions, and three-stage DEA modifies the efficiency. Then, we calculate the Malmquist-Luenberger index (TML) and its components: technological change (TTC) and technological efficiency change (TEC). Lastly, the influential factors impacting TSE are analyzed via a truncated regression Tobit model. The following are the conclusions: (1) The transportation industry in China exhibits inefficiency, and the average TSE in Stage I and III is 0.91 and 0.93, respectively. TSE is underestimated due to the influence of external environmental factors and inefficiencies in management in Stage I. (2) TSE in the eastern area also produces significant carbon emissions that surpass the national average. At the same time, other regions face efficiency limitations due to geographical constraints and management obstacles. (3) Insufficient technical capacity is a major cause of inefficiency in the transport sector and is prevalent in the northeast, west, and central regions. (4) Population growth and income per capita advancements foster transportation industry development, while increased GDP, fiscal revenues, and traffic accidents contribute to declining efficiency. The study above findings serve as a foundation for regional and national management initiatives and policies to enhance transportation effectiveness.

Gold perfusion experiments support the multi-layered, mesoporous nature of intervessel pit membranes in angiosperm xylem.

Fluid transport across intervessel pit membranes of angiosperm xylem plays a major role in plant transpiration, with transport resistance largely depending on pore constriction sizes. Traditionally, fluid particles traversing pit membranes are assumed to cross a single instead of multiple pore constrictions. We tested a multi-layered pit membrane model in xylem of eight angiosperm species by estimating the size frequency of pore constrictions in relation to pit membrane thickness and compared modelled data with perfusion characteristics of nanoscale gold particles based on transmission electron microscopy. The size frequency of modelled pore constrictions showed similar patterns to the measured number of perfused particle sizes inside pit membranes, although frequency values measured were 10-50 times below modelled data. Small particles enter pit membranes most easily, especially when injected in thin pit membranes. The trapping of gold particles by pore constrictions becomes more likely with increasing pore constriction number and pit membrane thickness. While quantitative differences between modelled and experimental data are due to various practical limitations, their qualitative agreement supports a multi-layered pit membrane model with multiple pore constrictions. Pore constrictions between 5 and 50 nm are realistic, and confirm the mesoporous nature of pit membranes.

Multilevel Pedicle Subtraction Osteotomy for Correction of Thoracolumbar Kyphosis in Ankylosing Spondylitis: Clinical Effect and Biomechanical Evaluation.

OBJECTIVE: To compare the clinical outcomes and biomechanical characteristics of 1-, 2-, and 3-level pedicle subtraction osteotomy (PSO), and establish selection criteria based on preoperative radiographic parameters. METHODS: Patients undergone PSO to treat ankylosing spondylitis from February 2009 to May 2019 in Sun Yat-sen Memorial Hospital of Sun Yat-sen University were enrolled. According to the quantity of osteotomy performed, the participants were divided into group A (1-level PSO, n = 24), group B (2-level PSO, n = 19), and group C (3-level PSO, n = 11). Clinical outcomes were assessed before surgery and at the final follow-up. Comparisons of the radiographic parameters and quality-of-life indicators were performed among and within these groups, and the selection criteria were established by regression. Finite element analysis was conducted to compare the biomechanical characteristics of the spine treated with different quantity of osteotomies under different working conditions. RESULTS: Three-level PSO improved the sagittal parameters more significantly, but resulted in longer operative time and greater blood loss (p < 0.05). Greater stress was found in the proximal screws and proximal junction area of the vertebra in the model simulating 1-level PSO. Larger stress of screws and vertebra was observed at the distal end in the model simulating 3-level PSO. CONCLUSION: Multilevel PSO works better for larger deformity correction than single-level PSO by allowing greater sagittal parameter correction and obtaining a better distribution of stress in the hardware construct, although with longer operation time and greater blood loss. Three-level osteotomy is recommended for the patients with preoperative of global kyphosis > 85.95°, T1 pelvic angle > 62.3°, sagittal vertical alignment > 299.55 mm, and pelvic tilt+ chin-brow vertical angle > 109.6°.

In vitro safety signals for potential clinical development of the anti-inflammatory pregnane X receptor agonist FKK6.

Based on the mimicry of microbial metabolites, functionalized indoles were demonstrated as the ligands and agonists of the pregnane X receptor (PXR). The lead indole, FKK6, displayed PXR-dependent protective effects in DSS-induced colitis in mice and in vitro cytokine-treated intestinal organoid cultures. Here, we report on the initial in vitro pharmacological profiling of FKK6. FKK6-PXR interactions were characterized by hydrogen-deuterium exchange mass spectrometry. Screening FKK6 against potential cellular off-targets (G protein-coupled receptors, steroid and nuclear receptors, ion channels, and xenobiotic membrane transporters) revealed high PXR selectivity. FKK6 has poor aqueous solubility but was highly soluble in simulated gastric and intestinal fluids. A large fraction of FKK6 was bound to plasma proteins and chemically stable in plasma. The partition coefficient of FKK6 was 2.70, and FKK6 moderately partitioned into red blood cells. In Caco2 cells, FKK6 displayed high permeability (A-B: 22.8 × 10-6 cm.s-1) and no active efflux. These data are indicative of essentially complete in vivo absorption of FKK6. The data from human liver microsomes indicated that FKK6 is rapidly metabolized by cytochromes P450 (t1/2 5 min), notably by CYP3A4. Two oxidized FKK6 derivatives, including DC73 (N6-oxide) and DC97 (C19-phenol), were detected, and these metabolites had 5-7 × lower potency as PXR agonists than FKK6. This implies that despite high intestinal absorption, FKK6 is rapidly eliminated by the liver, and its PXR effects are predicted to be predominantly in the intestines. In conclusion, the PXR ligand and agonist FKK6 has a suitable pharmacological profile supporting its potential preclinical development.

MicroRNA-18a regulates the Pyroptosis, Apoptosis, and Necroptosis (PANoptosis) of osteoblasts induced by tumor necrosis factor-α via hypoxia-inducible factor-1α.

BACKGROUND: Tumor necrosis factor-α (TNF-α) is involved in inflammatory responses and promotes cell death and the inhibition of osteogenic differentiation. MicroRNA (miRNA) plays a crucial role in the infected bone diseases, however, the biological role of miRNAs in inflammation-induced impaired osteogenic differentiation remains unclear. This study aimed to explore the role of miRNA-18a-5p (miR-18a) in regulating PANoptosis and osteogenic differentiation in an inflammatory environment via hypoxia-inducible factor-1α (HIF1-α). METHODS: The expression of miR-18a in MC3T3-E1 cells was analyzed using quantitative reverse transcription-polymerase chain reaction in an inflammatory environment induced by TNF-α. The expression of HIF1-α and NLRP3 in LV-miR-18a or sh-miR-18a cells was analyzed using western blotting. Fluorescence imaging for cell death, flow cytometry, and alkaline phosphatase activity analysis were used to analyze the role of miR-18a in TNF-α-induced PANoptosis and the inhibition of osteogenic differentiation. An animal model of infectious bone defect was established to validate the regulatory role of miR-18a in an inflammatory environment. RESULTS: The expression of miRNA-18a in the MC3T3-E1 cell line was significantly lower under TNF-α stimulation than in the normal environment. miR-18a significantly inhibited the expression of HIF1-α and NLRP3, and inhibition of HIF1-α expression further inhibited NLRP3 expression. Furthermore, inhibition of miR-18a expression promoted the TNF-α-induced PANoptosis and inhibition of osteogenic differentiation, whereas miR-18a overexpression and the inhibition of both HIF1-α and NLRP3 reduced the effects of TNF-α. These findings are consistent with those of the animal experiments. CONCLUSION: miRNA-18a negatively affects HIF1-α/NLRP3 expression, inhibits inflammation-induced PANoptosis, and impairs osteogenic differentiation. Thus, it is a potential therapeutic candidate for developing anti-inflammatory strategies for infected bone diseases.

Muscone ameliorates myocardial ischemia‒reperfusion injury by promoting myocardial glycolysis.

Objective: The incidence of acute myocardial infarction (AMI) is increasing yearly. With the use of thrombolysis or percutaneous coronary intervention (PCI), the mortality rate of acute myocardial infarction has been significantly reduced. However, reperfusion can cause additional myocardial injury. There is still a lack of effective drugs to treat I/R injury, and it is urgent to find new therapeutic drugs. Methods: In this study, network pharmacology was used to predict potential targets and biological processes involved in Muscone-mediated treatment of acute myocardial infarction. To model ischemia‒reperfusion injury, a hypoxia-reoxygenation model and in vivo ischemia‒reperfusion injury C57BL/6 mice model was constructed. Mice were treated with Muscone i.p. for 4 weeks. We detected the cardiac function on day 28.The expression levels of the apoptotic proteins Caspase-3 and Bax and the anti-apoptotic protein Bcl-2 were detected by immunoblotting after Muscone treatment of AC16 cells and in vivo. Additionally, the gene expression levels of the PUMA and p53 were analyzed by qRT‒PCR. Molecular docking was used to evaluate the binding energy between Muscone and NLRP3-related proteins. Immunoblotting and qRT‒PCR were used to assess the expression levels of NLRP3 signaling pathway-related proteins (NLRP3, ASC, and Caspase-1) and the NLRP3 gene, respectively. Moreover, the extracellular acidification rate of AC16 cells was measured using the Seahorse system to evaluate glycolysis levels after Muscone treatment. The expression of the key glycolytic enzyme PKM2 was analyzed by immunoblotting and qRT‒PCR. Finally, ChIP‒qPCR was performed to determine the levels of histone modifications (H3K4me3, H3K27me3, and H2AK119Ub) in the PKM2 promoter region. Results: GO functional enrichment analysis revealed that muscone was involved in regulating the biological processes (BP) of AMI, which mainly included negative regulation of the apoptosis signaling pathway, the response to lipopolysaccharide, and blood pressure regulation. The cellular components (CC) involved in muscone-mediated regulation of AMI mainly included lipid rafts, membrane microdomains, and membrane regions. The molecular functions (MF) involved in muscone-mediated regulation of AMI mainly included oxidoreductase activity, nuclear receptor activity, and transcription factor activity. In vitro results indicated that muscone treatment could inhibit the expression levels of Bax and Caspase-3 in AC16 cells after ischemia‒reperfusion while increasing the expression level of the antiapoptotic protein Bcl-2. Muscone significantly suppressed the transcription levels of p53 and PUMA in AC16 cells. Molecular docking suggested that muscone could bind well with the Cryo-EM structure of NEK7(PDB ID:6NPY). Further investigation of inflammatory pathways revealed that muscone could inhibit the expression level of NLRP3 in AC16 cells and reduce the expression levels of Caspase-1 and Caspase recruitment domain. Fluorescent quantitative PCR experiments showed that muscone significantly inhibited the transcription of NLRP3. Moreover, we found that muscone could enhance the glycolytic efficiency of AC16 cells, which may be related to the increased protein expression of PKM2 in AC16 cells. Fluorescent quantitative PCR showed that muscone could increase the transcription level of PKM2. Chromatin immunoprecipitation assays showed that muscone treatment increased the expression level of H3K4me3 in the PKM2 promoter region and inhibited the levels of H3K27me3 and H2AK119Ub in the PKM2 promoter region. Conclusion: Muscone promoted myocardial glycolysis and inhibited NLRP3 pathway activation to improve myocardial ischemia‒reperfusion injury.

The impact of population agglomeration on ecological resilience: Evidence from China.

Due to climate change and human activities, ecological and environmental issues have become increasingly prominent and it is crucial to deeply study the coordinated development between human activities and the ecological environment. Combining panel data from 31 provinces in China spanning from 2011 to 2020, we employed a fixed-effects model, a threshold regression model, and a spatial Durbin model to empirically examine the intricate impacts of population agglomeration on ecological resilience. Our findings indicate that population agglomeration can have an impact on ecological resilience and this impact depends on the combined effects of agglomeration and crowding effects. Also, the impact of population agglomeration on ecological resilience exhibits typical dual-threshold traits due to differences in population size. Furthermore, population agglomeration not only directly impacts the ecological resilience of the local area, but also indirectly affects the ecological resilience of surrounding areas. In conclusion, we have found that population agglomeration does not absolutely impede the development of ecological resilience. On the contrary, to a certain extent, reasonable population agglomeration can even facilitate the progress of ecological resilience.