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Abstract Background Currently, the synthesis of compounds based on microbial cell factories is rapidly advancing, yet it encounters several challenges. During the production process, engineered strains frequently encounter disturbances in the cultivation environment or the impact of their metabolites, such as high temperature, acid-base imbalances, hypertonicity, organic solvents, toxic byproducts, and mechanical damage. These stress factors can constrain the efficiency of microbial fermentation, resulting in slow cell growth, decreased production, significantly increased energy consumption, and other issues that severely limit the application of microbial cell factories. Results This study demonstrated that sterol engineering in Kluyveromyces marxianus, achieved by overexpressing or deleting the coding genes for the last five steps of ergosterol synthase (Erg2-Erg6), altered the composition and ratio of sterols in its cell membrane, and affected its multiple tolerance. The results suggest that the knockout of the Erg5 can enhance the thermotolerance of K. marxianus, while the overexpression of the Erg4 can improve its acid tolerance. Additionally, engineering strain overexpressed Erg6 improved its tolerance to elevated temperature, hypertonic, and acid. YZB453, obtained by overexpressing Erg6 in an engineering strain with high efficiency in synthesizing xylitol, produced 101.22 g/L xylitol at 45oC and 75.11 g/L xylitol at 46oC. Using corncob hydrolysate for simultaneous saccharification and fermentation (SSF) at 46oC that xylose released from corncob hydrolysate by saccharification with hemicellulase, YZB453 can produce 45.98 g/L of xylitol, saving 53.72% of the cost of hemicellulase compared to 42oC. Conclusions This study elucidates the mechanism by which K. marxianus acquires resistance to various antifungal drugs, high temperatures, high osmolarity, acidity, and other stressors, through alterations in the composition and ratio of membrane sterols. By employing sterol engineering, the fermentation temperature of this unconventional thermotolerant K. marxianus was further elevated, ultimately providing an efficient platform for synthesizing high-value-added xylitol from biomass via the SSF process at temperatures exceeding 45 °C.

Abstract Protein-protein interactions (PPIs) stabilization with molecular glues plays a crucial role in drug discovery, albeit with significant challenges. In this study, we propose a dual-site approach, targeting the PPI region and its dynamic surroundings. We conduct molecular dynamics simulations to identify critical sites on the PPI that stabilize the cyclin-dependent kinase 12 - DNA damage-binding protein 1 (CDK12-DDB1) complex, resulting in further cyclin K degradation. This exploration leads to the creation of LL-K12-18, a dual-site molecular glue, which enhances the glue properties to augment degradation kinetics and efficiency. Notably, LL-K12-18 demonstrates strong inhibition of gene transcription and anti-proliferative effects in tumor cells, showing significant potency improvements in MDA-MB-231 (88-fold) and MDA-MB-468 cells (307-fold) when compared to its precursor compound SR-4835. These findings underscore the potential of dual-site approaches in disrupting CDK12 function and offer a structural insight-based framework for the design of cyclin K molecular glues.

BaiYangJie (BYJ) is a terrestrial perennial plant commonly used as a Dai medicine and has therapeutic effects on liver and kidney diseases. Cisplatin (CP), a chemotherapy drug, has good therapeutic effects but causes many side effects, including nephrotoxicity. This article investigated the toxicology of the methanol extract of BYJ (ME-BYJ) and its protective effect on CP-induced acute kidney injury (AKI) through pharmacological experiments. The results showed that the treated mice had no toxicological symptoms and no anatomical, physiological, or histological abnormalities. The BYJ-high-dose group showed significantly attenuated CP-induced AKI. It is concluded that ME-BYJ has the most significant protective effect on AKI at a dose of 8 g/kg and BYJ was not toxic.

Abstract To determine the presence of a consistent osseous corridor from the lateral-posterior aspect of the anterior inferior iliac spine to the sacral wing that could be used for safe trans percutaneous screw fixation for pelvic fragility fractures of the iliac wing and fracture dislocations of the sacroiliac joint (FFP types IIIa and IIIb). Computed tomography (CT) scans were obtained from 100 patients and imported to Mimics software for 3D reconstruction. Then, a cylinder was drawn to imitate the modified LC-II screw and adjusted to a maximum radius and length to obtain the feasible region. Thirteen parameters of the osseous corridor of the modified LC-II screw were measured. Differences between sex groups were compared, and significant statistical correlations were carefully studied to determine potentially important clinical relationships. The records of patients with FFP type IIIa and IIIb fragility fractures of the pelvis were extracted from our hospital. Patients who underwent modified LC-II screw fixation, LC-II screw fixation or reconstruction plate fixation were included. Patients’ operative characteristics and complications were recorded at follow-up. Fracture reduction quality was assessed using the Matta standard. Functional outcomes were evaluated using the Majeed grading system. The mean maximum diameters of the osseous corridors of the modified LC-II screw in males and females were 12.73 and 10.83 mm, respectively. The mean maximum lengths of the osseous corridors of the modified LC-II screw in males and females were 96.37 and 93.37 mm, respectively. In the treatment of patients with FFP IIIa and FFP IIIb fractures, the group of treatment by the modified LC-II screws fixation was shown significantly shorter operative time and fewer intraoperative blood loss in comparison to that by the reconstruction plates. In the present study, all the males and females had a complete osseous corridor of the modified LC-II screw. The clinical results of the patients who were treated with modified LC-II screw fixation suggest that the novel method has a good preliminary outcome.

Background/Aims Metabolic dysfunction-associated steatotic liver disease (MASLD) has become an increasingly important health challenge, with a substantial rise linked to changing lifestyles and global obesity. Ursolic acid, a natural pentacyclic triterpenoid, has been explored for its potential therapeutic effects. Given its multifunctional bioactive properties, this research further revealed the pharmacological mechanisms of ursolic acid on MASLD. Methods Drug target chips and bioinformatics analysis were combined in this study to explore the potential therapeutic effects of ursolic acid on MASLD. Molecular docking simulations, surface plasmon resonance analyses, pull-down experiments, and co-immunoprecipitation assays were used to verify the direct interactions. Gene knockdown mice were generated, and high-fat diets were used to validate drug efficacy. Furthermore, initial CD4+ T cells were isolated and stimulated to demonstrate our findings. Results In this study, the multifunctional extracellular matrix phosphorylated glycoprotein secreted phosphoprotein 1 (SPP1) was investigated, highlighting its capability to induce Th17 cell differentiation, amplifying inflammatory cascades, and subsequently promoting the evolution of MASLD. In addition, this study revealed that in addition to the canonical TGF-β/IL-6 cytokine pathway, SPP1 can directly interact with ITGB1 and CD44, orchestrating Th17 cell differentiation via their joint downstream ERK signaling pathway. Remarkably, ursolic acid intervention notably suppressed the protein activity of SPP1, suggesting a promising avenue for ameliorating the immunoinflammatory trajectory in MASLD progression. Conclusions Ursolic acid could improve immune inflammation in MASLD by modulating SPP1-mediated Th17 cell differentiation via the ERK signaling pathway, which is orchestrated jointly by ITGB1 and CD44, emerging as a linchpin in this molecular cascade.

Objective To compare the clinical efficacy of anterior column reconstruction using single or double titanium mesh cage (TMC) after total en bloc spondylectomy (TES) of thoracic and lumbar spinal tumors. Methods A retrospective cohort study was performed involving 39 patients with thoracic or lumbar spinal tumors. All patients underwent TES, followed by anterior reconstruction and screw-rod instrumentation via a posterior-only procedure. Twenty-two patients in group A were treated with a single TMC to reconstruct the anterior column, whereas 17 patients in group B were reconstructed with double TMCs. Results The overall follow-up is 20.5 ± 4.6 months. There is no significant difference between the 2 groups regarding age, sex, body mass index, tumor location, operative time, and intraoperative blood loss. The time for TMC placement was significantly shortened in the double TMCs group (5.2 ± 1.3 minutes vs. 15.6 ± 3.3 minutes, p = 0.004). Additionally, postoperative neural complications were significantly reduced with double TMCs (5/22 vs. 0/17, p = 0.046). The kyphotic Cobb angle and mean intervertebral height were significantly corrected in both groups (p ≤ 0.001), without obvious loss of correction at the last follow-up in either group. The bone fusion rates for single TMC and double TMCs were 77.3% and 76.5%, respectively. Conclusion Using 2 smaller TMCs instead of a single large one eases the placement of TMC by shortening the time and avoiding nerve impingement. Anterior column reconstruction with double TMC is a clinically feasible, and safe alternative following TES for thoracic and lumbar tumors.

Abstract Although the chloroplast genome (cpDNA) of higher plants is known to exist as a large protein-DNA complex called ‘plastid nucleoid’, researches on its DNA state and regulatory elements are limited. In this study, we performed the assay for transposase-accessible chromatin sequencing (ATAC-seq) on five common tissues across five grasses, and found that the accessibility of different regions in cpDNA varied widely, with the transcribed regions being highly accessible and accessibility patterns around gene start and end sites varying depending on the level of gene expression. Further analysis identified a total of 3970 putative protein binding footprints on cpDNAs of five grasses. These footprints were enriched in intergenic regions and co-localized with known functional elements. Footprints and their flanking accessibility varied dynamically among tissues. Cross-species analysis showed that footprints in coding regions tended to overlap non-degenerate sites and contain a high proportion of highly conserved sites, indicating that they are subject to evolutionary constraints. Taken together, our results suggest that the accessibility of cpDNA has biological implications and provide new insights into the transcriptional regulation of chloroplasts.

Abstract. Background:. The ability to generate functional hepatocytes without relying on donor liver organs holds significant therapeutic promise in the fields of regenerative medicine and potential liver disease treatments. Clustered regularly interspaced short palindromic repeats (CRISPR) activator (CRISPRa) is a powerful tool that can conveniently and efficiently activate the expression of multiple endogenous genes simultaneously, providing a new strategy for cell fate determination. The main purpose of this study is to explore the feasibility of applying CRISPRa for hepatocyte reprogramming and its application in the treatment of mouse liver fibrosis. Method:. The differentiation of mouse embryonic fibroblasts (MEFs) into functional induced hepatocyte-like cells (iHeps) was achieved by utilizing the CRISPRa synergistic activation mediator (SAM) system, which drove the combined expression of three endogenous transcription factors—Gata4, Foxa3, and Hnf1a—or alternatively, the expression of two transcription factors, Gata4 and Foxa3. In vivo, we injected adeno-associated virus serotype 6 (AAV6) carrying the CRISPRa SAM system into liver fibrotic Col1a1-CreER; Cas9fl/fl mice, effectively activating the expression of endogenous Gata4 and Foxa3 in fibroblasts. The endogenous transcriptional activation of genes was confirmed using real-time quantitative polymerase chain reaction (RT-qPCR) and RNA-seq, and the morphology and characteristics of the induced hepatocytes were observed through microscopy. The level of hepatocyte reprogramming in vivo is detected by immunofluorescence staining, while the improvement of liver fibrosis is evaluated through Sirius red staining, alpha-smooth muscle actin (α-SMA) immunofluorescence staining, and blood alanine aminotransferase (ALT) examination. Results:. Activation of only two factors, Gata4 and Foxa3, via CRISPRa was sufficient to successfully induce the transformation of MEFs into iHeps. These iHeps could be expanded in vitro and displayed functional characteristics similar to those of mature hepatocytes, such as drug metabolism and glycogen storage. Additionally, AAV6-based delivery of the CRISPRa SAM system effectively induced the hepatic reprogramming from fibroblasts in mice with live fibrosis. After 8 weeks of induction, the reprogrammed hepatocytes comprised 0.87% of the total hepatocyte population in the mice, significantly reducing liver fibrosis. Conclusion:. CRISPRa-induced hepatocyte reprogramming may be a promising strategy for generating functional hepatocytes and treating liver fibrosis caused by hepatic diseases.

Methane (CH4) clumped isotope analysis plays a crucial role in the fields of climate change, energy exploration, and planetary research. The purity of CH4 in samples directly affects the precision and accuracy of high-resolution mass spectrometry in clumped isotope analysis. Addressing the challenge associated with enriching and purifying CH4 components in gas samples, this study optimized conditions such as carrier gas line speed and sample injection volume based on the principles of gas chromatography (GC) component separation, with real-time monitoring of component peak shapes. Additionally, the recovery rate was quantified using an external standard method and purity was verified through GC component analysis to ensure the effectiveness of the purification process. By optimizing the chromatography-vacuum low-temperature enrichment preparation method, the optimal carrier gas line speed for the IBEX system was determined to be 12 mL/min, with a CH4 injection volume less than 12 mL. This facilitated visualization of GC peak shapes, thus ensured that the CH4 peak was essentially separated from the adjacent N2 interference peak, achieving high-purity enrichment of the CH4 single component. When the CH4 content in gas samples was less than 70% and the air content was high, secondary purification was required to improve CH4 purity. The causes of CH4 isotopic fractionation during purification using adsorbents like 5Å molecular sieves were discussed, and extending the CH4 collection time was proposed to eliminate the interference from the 5Å molecular sieve. Currently, this method requires approximately 90 min for a single purification process, with CH4 recovery and purity ranging from 90.1% to 95.7% and 97.3% to 98.9%, respectively. The differences in isotopic composition (δ13CVPDB and δDVSMOW, Δ13CH3D, and Δ12CH2D2) are all less than the analytical error of the mass spectrometer, making them almost negligible.

Abstract Scar tissue is the inevitable result of repairing human skin after it has been subjected to external destructive stimuli. It leads to localized damage to the appearance of the skin, accompanied by symptoms such as itching and pain, which reduces the quality of life of the patient and causes serious medical burdens. With the continuous development of economy and society, there is an increasing demand for beauty. People are looking forward to a safer and more effective method to eliminate pathological scarring. In recent years, adipose-derived stem cells (ADSCs) have received increasing attention from researchers. It can effectively improve pathological scarring by mediating inflammation, regulating fibroblast proliferation and activation, and vascular reconstruction. This review focuses on the pathophysiological mechanisms of hypertrophic scarring, summarizing the therapeutic effects of in vitro, in vivo, and clinical studies on the therapeutic effects of ADSCs in the field of hypertrophic scarring prevention and treatment, the latest application techniques, such as cell-free therapies utilizing ADSCs, and discussing the advantages and limitations of ADSCs. Through this review, we hope to further understand the characterization of ADSC and clarify the effectiveness of its application in hypertrophic scarring treatment, so as to provide clinical guidance.

Abstract Radiated tumor cell‐derived extracellular vesicles (RT‐EVs) encapsulate abundant DNA fragments from irradiated tumor cells, in addition to acting as integrators of multiple tumor antigens. Accumulating evidence indicates these DNA fragments from damaged cells are involved in downstream immune responses, but most of them are degraded in cells before incorporation into derived RT‐EVs, thus the low abundance of DNA fragments limits immune responses of RT‐EVs. Here, this study found that different radiations affected fates of DNA fragments in RT‐EVs. Boron neutron capture therapy (BNCT) induced DNA accumulation in RT‐EVs (BEVs) by causing more DNA breaks and DNA oxidation resisting nuclease degradation. This is attributed to the high‐linear energy transfer (LET) properties of alpha particles from the neutron capture reaction of 10B. When being internalized by dendritic cells (DCs), BEVs activated the DNA sensing pathway, resulting in functional enhancements including antigen presentation, migration capacity, and cytokine secretion. After vaccination of the BEVs‐educated DCs (BEV@BMDCs), the effector T cells significantly expanded and infiltrated into tumors, suggesting robust anti‐tumor immune activation. BEV@BMDCs not only effectively inhibited the primary tumor growth and metastasis formation but also elicited long‐term immune memory. In conclusion, a successful DC vaccine is provided as a promising candidate for tumor vaccine.

The nano and micro diamond film coated tools were prepared by the hot wire CVD method.The surface morphology of the film was characterized by field emission scanning electron microscopy. The prepared CVD diamond coated tools were used to end mill the aluminum alloy surface forward and backward at high speed under the condition of dry cutting without lubrication. It studies the characteristics and size of edge burrs during high-speed end milling of aluminum alloy using CVD diamond coated tools. It carries out orthogonal tests on cutting process parameters to explore the cutting parameters and processes with little or no cutting burr when nano-diamonds coated tools are used for high-speed forward milling. The results show that the distribution of burrs on the edges of the workpiece is uneven after milling, and the burrs during forward milling are sparse and small in size. Among them, the average height of burrs on the edges during forward milling with nano diamond coated tools is 32.08 μm, only 46.5% of the forward milling burr height of micrometer diamond coated tools. When nano diamond coated tools are used for high-speed forward milling of flat surfaces, vc has the greatest impact on edge burrs, followed by vf, and ae has the weakest impact. The optimal parameter combination for high-speed milling is ae=4 mm, vf=2 000 mm/min, and vc=400 m/min. The average burr height after milling is 21.29 μm. When the diamond coated tool is used to end mill the aluminum alloy plane, in order to obtain a small burr, the nano diamonds coated tool is selected, the cutting method of forward milling and the corresponding high-speed cutting parameters are adopted.

Metal halide perovskite has generated significant attention due to its high optical absorption coefficient, the tunability of bandgap, and its solution processing properties, all of which have led to a range of applications including solar cells, photodetectors, light-emitting diodes and lasers. However, the material's application as an optical modulator for high-energy ultrafast lasers is still limited by its slow nonlinear optical response, low damage threshold and the inferior stability. Herein, we developed a saturable absorber (SA) with the thermally stable Cs0.15FA0.85PbI2.85Br0.15 perovskite thin films as an optical modulator towards the near-infrared nanosecond passively Q-switched Nd:YAG laser generation for the first time. The Cs0.15FA0.85PbI2.85Br0.15 perovskite SA optical modulator produced an exceptional nonlinear optical response with a notable modulation depth of 15.1%, an ultrashort recovery time of 52.46 ps, and a high optical damage threshold exceeding 275.9 mJ/cm2. Of further significance, it displayed an excellent operational stability, and when incorporated into the oscillator of the Nd:YAG laser, allowed for a stable operation of 166 ns pulse laser with a center wavelength of 1.06 μm. The findings provide a pathway for the employment of the Cs0.15FA0.85PbI2.85Br0.15 perovskite SA optical modulator in near-infrared ultrafast laser applications.

Background: There is a strong causal relationship between intimate partner violence and major depressive disorder, which partly endangers women's safety across the life course and potentially affects the development of future generations. The international community has placed a high priority on addressing the intimate partner violence and the resulting burden of mental illness. Data collection needs to be captured across the temporal trend and spatial distribution for major depressive disorder attributed to intimate partner violence, to reflect the priorities and expectations of survivors.Method: This research obtained raw disability-adjusted life years (DALYs) information for major depressive disorder attributed to intimate partner violence from the Global Burden of Disease 2019. Using estimated annual percentage change and two-way fixed effects models, a secondary spatio-temporal analysis of the age-standardized DALYs rate from 1990 to 2019 was performed.Results: In 2019, DALYs lost among women experiencing major depressive disorder (3.16 million) accounted for 37.18% of the DALYs lost worldwide due to intimate partner violence. The age-standardized DALYs rate of major depressive disorder attributed to intimate partner violence was 108.57 per 100,000. The highest was concentrated in the menopausal transition (45-55), with 133.61 per 100,000, and particularly distributed in Uganda (429.31 per 100,000). The early reproductive period (15–19) showed the increasing age-standardized DALYs rate from 1990 to 2019, which was mainly driven by Malaysia (3.73% per year). Furthermore, countries with higher initial levels of the age-standardized DALYs rate were growing faster than those with lower levels.Conclusions: The burden of major depressive disorder attributed to intimate partner violence showed biological and spatial inequality, prioritized intervention should be targeted at vulnerable stage women in their early reproductive period and menopausal transition. Combined political, socio-cultural as well as medical measures to prevent violence and treat major depressive disorder should be implemented and developed.

The gut microbiota plays an important role in the development and treatment of hepatocellular carcinoma (HCC). However, the implication of specific gut microbiota in targeted sorafenib therapy for advanced HCC and the microbiota mode of action, remain to be elucidated. Here, we confirmed that four bacterial genera, Lachnoclostridium, Lachnospira, Enterobacter and Enterococcus, are associated with the therapeutic efficacy of Sorafenib, and that Enterobacter faecium (Efm) plays a crucial role in modulating the sorafenib activity. The effective colonization by Emf induced the IL-12 and IFN-γ production and an increased proportion of IFN-γ+CD8+ T cells in the tumor microenvironment. Finally, exopolysaccharides (EPS) from Efm were the primary inducer to prompt IFN-γ+CD8+ T cells to secrete IFN-γ, which together with sorafenib instigated ferroptosis in HCC cells. Collectively, these results indicate that Efm is a promising probiotics that enhances the efficacy of sorafenib treatment in advanced HCC.

BackgroundMitochondrial dysfunction has been shown to play a critical role in cancer biology. However, its involvement in intrahepatic cholangiocarcinoma (iCCA) remains significantly understudied.MethodsRNA sequencing data of 30 pairs of iCCA and paracancerous tissues were collected from the First Affiliated Hospital of Wenzhou Medical University (WMU). The WMU cohort (n = 30) was integrated with public TCGA (n = 30) and GSE107943 (n = 30) datasets to establish a multi-center iCCA cohort. We merged the TCGA and GSE107943 cohorts into an exploration cohort to develop a mitochondria signature for prognosis assessment, and utilized the WMU cohort for external validation. Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and Hallmarker analyses were used for functional interpretation of iCCA associated mitochondria-related genes (MRGs). In addition, unsupervised clustering was performed to identify mitochondria-based iCCA subtypes with the data of three institutions. Further investigations were conducted to examine the impact of mitochondrial dysfunction on drug responses, alteration of the tumor immune microenvironment, and immune responses.ResultsTwo hundred and sixty-three iCCA-related MRGs were identified to be related to fatty acid metabolism, oxidative phosphorylation, and apoptosis. Through univariate and multivariate Cox, and LASSO analyses, a mitochondria signature with five optimal MRGs was established to evaluate the prognosis of iCCA patients with the AUC values ranged from 0.785 to 0.928 in the exploration cohort. The signature also exhibited satisfactory performance in the WMU cohort with AUC values of 0.817–0.871, and was identified as an independent risk predictor in both cohorts. Additionally, we found that patients with higher mitochondria score with poor prognosis presented lower infiltration levels of CD4+ T-cell, NK cells, and monocytes, and demonstrated higher sensitivity to targeted therapies, including sorafenib. Furthermore, two distant mitochondria-based subtypes were determined, and subtype 2 was associated with shorter survival time and immunosuppressive tumor microenvironment. Finally, the differential protein expression of five key MRGs was verified by Immunohistochemistry.ConclusionWe found mitochondrial dysfunction modulates aberrant metabolism, oxidative stress, immune responses, apoptosis, and drug sensitivity in iCCA. A mitochondria signature and two mitochondria-based iCCA subtypes were identified for clinical risk stratification and immunophenotyping.

Vibrio harveyi is a major pathogen in yellow drum (Nibea albiflora) aquaculture, causing significant mortality and economic losses. In this study, using the latest assembled reference genome of yellow drum by our laboratory, we conducted genome-wide association study (GWAS) analysis on 345 individuals (197 susceptible and 148 resistant). The analysis revealed 24 significant single nucleotide polymorphisms (SNPs) on chromosome 24 within a 217 Kb region. The estimated heritability for all genome-wide SNPs was 0.3578, while the heritability for the 24 significant SNPs was 0.0710. Four candidate genes were identified within this region: Suppressor of Cytokine Signaling 1 (SOCS1), C-type Lectin Domain Family 16A (CLEC16A), Major Histocompatibility Complex Class II Transactivator (CIITA), and Protein Kinase Cβ (PRKCB). Subsequently, expression quantitative trait loci (eQTL) analysis was performed on transcriptome sequencing data from spleen tissues of 78 individuals from the resistant group. On average, each chromosome harbored 49,396 eQTL loci, with an average of one SNP regulate 1.3 genes. Notably, 22.79% of SNPs showed significant associations with the expression of one or more genes. By integrating GWAS and eQTL data, seven SNPs were identified to have significant associations with regulated genes in the eQTL results. All seven SNPs were found to target the same gene, namely Zinc Finger Protein yd23210 in yellow drum. This study provides genetic markers and candidate genes for molecular breeding of yellow drum against V. harveyi infection, offering insights into the molecular immune mechanisms and potential pathways for genetic improvement of disease resistance traits in this species.

The aim of this paper is to study the flexural behavior of reinforced concrete (RC) beams reinforced by engineered cementitious composite (ECC). The reinforcement effect of ECC on RC beam is studied by experiments, numerical simulation and theoretical analysis, considering the secondary load. Five beams were tested under static load, including two beams with ECC layers at the top and bottom (TBEB), two beams with ECC layers at the bottom (BEB) and one unreinforced beam. Different parameters in the test include reinforcement ratio, whether the top is equipped with ECC layer, and the thickness of the top ECC layer. The test data were used to validate the finite element model developed to simulate the bending response of the ECC layer reinforced beam. Then, a series of parameter studies were carried out to evaluate the effects of ECC layer thickness, reinforcement ratio and ECC elastic modulus on the reinforced beam. The experimental and numerical simulation results show that compared with other reinforced beams, TBEB equipped with 50 mm ECC layer at the top and bottom has better performance. Compared with the control beams, the ultimate bearing capacity is increased by 82.6 %, ductility and deformation capacity are increased by 2.38 times and 2.11 times, respectively. Finally, a calculation model for predicting the flexural capacity of TBEB is presented. The predicted flexural capacity is in good agreement with the experimental results.

In geotechnical engineering, side friction resistance (SFR) is difficult to be measured directly. To further understand distribution law of the SFR, this paper developed a monitoring device that can directly measure the SFR. Further, a theoretical conversion formula for the elastic deformation and the SFR that considers the end effect of sensor sealing was proposed to guide the selection of sensor size and sealing material. Moreover, the monitoring device for the SFR was then calibrated using a large-scale direct shear apparatus and analyzed the stability of the sensor. The calibration results revealed that under cyclic loading and unloading conditions, the linear correlation coefficient of the sensor was greater than 0.996, and the sensitivity after sealing could reach 4.836 με/kPa, which met requirements of the engineering application. The developed monitoring device characterized by simple testing principle, low cost, and high precision were successfully applied to an open caisson project in Harbin City, which contributes to address the difficult problem of efficiently collecting the SFR in highways, bridges, water conservancy, and other projects.

BackgroundLaminotomy and laminar replantation have emerged as novel treatment modalities for intraspinal tumors, aiming to minimize postoperative complications and retain spinal mobility. However, existing research predominantly emphasizes their application in the thoracolumbar spine. The unique anatomy of the atlantoaxial segments necessitates surgical techniques that differ from those used in other spinal regions, and the clinical effect of such procedure remains unknown.Case presentationA 61-year-old male patient with intradural schwannoma at the atlantoaxial level was operated on. The patient underwent posterior laminectomy, as well as a combined replantation of the posterior arch of the atlas and bilateral axial laminae. Postoperatively, the patient experienced significant neurological improvement, with no deformities or instability on the radiological assessments during the follow-up.ConclusionLaminotomy with combined replantation of the posterior arch of the atlas and bilateral axial lamina emerges as an effective approach for managing intraspinal tumors at the atlantoaxial level. This technique not only offers ample operating space but also restores the stability of the spinal canal. Moreover, it preserves the mobility of the atlantoaxial segment, minimizes impact on adjacent segments, and mitigates the formation of postoperative fibrosis.

Background: Constipation is one of the chronic gastrointestinal functional diseases that affects the quality of life. While Qi Lang Formula (QLF) has demonstrated effectiveness in alleviating constipation symptoms, its precise mechanism remains elusive. Methods: QLF was analyzed using UPLC-MS/MS. Targets for QLF were collected from SwissADME, Herb, ITCM databases, and constipation-related targets from scRNA-seq and Genecards databases. Overlapping targets suggested potential QLF therapy targets for constipation. Enrichment analysis used the KOBAS database. A ''drug-ingredient-target'' network was constructed with Cytoscape, and AutoDock verified active ingredient binding. H&E staining assessed colonic mucosa changes, TEM examined ICC structural changes. ELISA measured neurotransmitter levels, and Western blot verified QLF's effect on target proteins. ICC proliferation was observed through immunofluorescence. Results: We identified 89 targets of QLF associated with ICC-related constipation, with c-Kit emerging as the pivotal target. Molecular docking studies revealed that Atractylenolide Ⅲ, Apigenin, Formononetin, Isorhamnetin, Naringenin, and Ononin exhibited strong binding affinities for the c-Kit structural domain. QLF significantly enhanced first stool passage time, fecal frequency, fecal moisture content, and intestinal propulsion rate. Further analysis unveiled that QLF not only restored neurotransmitter levels but also mitigated colon muscular fiber ruptures. ICC ultrastructure exhibited partial recovery, while Western blot confirmed upregulated c-Kit expression and downstream targets. Immunofluorescence results indicated ICC proliferation post QLF treatment in rat colon. Conclusion: Our findings suggest that QLF may promote ICC proliferation by targeting SCF/c-Kit and its downstream signaling pathway, thereby regulating intestinal motility.

Abstract Background Recent advances in gene editing technology have opened up new avenues for in vivo gene therapy, which holds great promise as a potential treatment method for dilated cardiomyopathy (DCM). The CRISPR-Cas13 system has been shown to be an effective tool for knocking down RNA expression in mammalian cells. PspCas13b, a type VI-B effector that can be packed into adeno-associated viruses and improve RNA knockdown efficiency, is a potential treatment for diseases characterized by abnormal gene expression. Results Using PspCas13b, we were able to efficiently and specifically knockdown the mutant transcripts in the AC16 cell line carrying the heterozygous human TNNT2 R141W (hTNNT2 R141W ) mutation. We used adeno-associated virus vector serotype 9 to deliver PspCas13b with specific single guide RNA into the hTNNT2 R141W transgenic DCM mouse model, effectively knocking down hTNNT2 R141W transcript expression. PspCas13b-mediated knockdown significantly increased myofilament sensitivity to Ca2+, improved cardiac function, and reduced myocardial fibrosis in hTNNT2 R141W DCM mice. Conclusions These findings suggest that targeting genes through Cas13b is a promising approach for in vivo gene therapy for genetic diseases caused by aberrant gene expression. Our study provides further evidence of Cas13b’s application in genetic disease therapy and paves the way for future applicability of genetic therapies for cardiomyopathy.

Abstract Aims Neutrophil extracellular traps (NETs) have been implicated in thrombotic diseases. There is no definitive explanation for how NETs form during acute ischemic strokes (AIS). The purpose of our study was to investigate the potential mechanism and role of NETs formation in the AIS process. Methods As well as 45 healthy subjects, 45 patients with AIS had ELISA tests performed to detect NET markers. Expression of high-mobility group box 1 (HMGB1) on platelet microvesicles (PMVs) was analyzed by flow cytometry in healthy subjects and AIS patients’ blood samples. We established middle cerebral artery occlusion (MCAO) mice model to elucidate the interaction between PMPs and NETs. Results A significant elevation in NET markers was found in patient plasma in AIS patients, and neutrophils generated more NETs from patients’ neutrophils. HMGB1 expression was upregulated on PMVs from AIS patients and induced NET formation. NETs enhanced Procoagulant activity (PCA) through tissue factor and via platelet activation. Targeting lactadherin in genetical and in pharmacology could regulate the formation of NETs in MCAO model. Conclusions NETs mediated by PMVs derived HMGB1 exacerbate thrombosis and brain injury in AIS. Video Abstract

The environmental agenda is decisive in the formation of the world economy at the present stage. The need for a green transition is rarely questioned by scientists. However, there remains a wide range of opinions about which instruments are the most effective in achieving the set goals of reducing the carbon footprint. The most popular mechanisms today are the carbon market and carbon tax mechanisms. Aim of the research was using modeling, literature analysis, and deduction methods to compare the similarities and differences between the basic principles of applying a carbon tax and a carbon market mechanism, the authors of the article identify the advantages and disadvantages of both approaches. The results of the research showed that the carbon market mechanism has obvious advantages such as flexibility, cross-industry and crossterritory coverage, and relative benefits for companies compared to a carbon tax mechanism. At the same time, the carbon tax mechanism has the obvious advantage of low management costs and wide coverage, but, on the other hand, these advantages are offset by potential inefficiencies and high “cost” for companies. The analysis of the policies of large countries with large carbon emissions over the past 2-3 years shows that they are more likely to introduce elements of the carbon market to reduce emissions. Based on the theoretical analysis of the main characteristics of the carbon market and carbon tax, as well as their advantages and disadvantages, the authors urge not to be limited to one of the options, but to use both mechanisms simultaneously. In particular, the carbon market should be the main mechanism when the goal is to create more incentives to actively reduce carbon emissions, while carbon tax policies can be an additional measure to incentivize or “punish” those enterprises that exceed emission standards, also, the carbon tax will help to involve those industries that, for various reasons, cannot be represented on the carbon market.

Abstract Background Brain metastases (BMs) are the most frequent intracranial tumours associated with poor clinical outcomes. Radiotherapy is essential in the treatment of these tumours, although the optimal radiation strategy remains controversial. The present study aimed to assess whether whole brain radiation therapy with a simultaneous integrated boost (WBRT + SIB) provides any therapeutic benefit over WBRT alone. Methods We included and retrospectively analysed 82 patients who received WBRT + SIB and 83 who received WBRT alone between January 2012 and June 2021. Intracranial progression-free survival (PFS), local tumour control (LTC), overall survival (OS), and toxicity were compared between the groups. Results Compared to WBRT alone, WBRT + SIB improved intracranial LTC and PFS, especially in the lung cancer subgroup. Patients with high graded prognostic assessment score or well-controlled extracranial disease receiving WBRT + SIB had improved intracranial PFS and LTC. Moreover, WBRT + SIB also improved the long-term intracranial tumour control of small cell lung cancer patients. When evaluating toxicity, we found that WBRT + SIB might slightly increase the risk of radiation-induced brain injury, and that the risk increased with increasing dosage. However, low-dose WBRT + SIB had a tolerable radiation-induced brain injury risk, which was lower than that in the high-dose group, while it was comparable to that in the WBRT group. Conclusions WBRT + SIB can be an efficient therapeutic option for patients with BMs, and is associated with improved intracranial LTC and PFS. Furthermore, low-dose WBRT + SIB (biologically effective dose [BED] ≤ 56 Gy) was recommended, based on the acceptable risk of radiation-induced brain injury and satisfactory tumour control. Trial registration Retrospectively registered.

Hand-foot syndrome (HFS) is a widely recognized dose-limiting cutaneous toxicity effect of fluoropyrimidine chemotherapy agents that impairs clinical benefits and treatment outcomes. Even though the cause and pathophysiology of HFS are relatively widely reported, how the toxicity of fluoropyrimidine translates into persistent inflammation has not been studied. Additionally, prevention and treatment strategies for HFS based on its mechanistic occurrence and development are scarce. In our study, we demonstrated that cGAS-STING signaling pathway-mediated cellular senescence played a critical role in the inflammatory reaction and provided a therapeutic solution for HFS. Mechanistically, DNA damage, as the primary cytotoxic cause, in keratinocytes induces cell cycle arrest, activates the cGAS-STING signaling pathway, and subsequently mediates cellular senescence, ultimately fueling a robust secondary inflammatory response that results in HFS. More importantly, the thymidine prodrug thymidine diacetate was proven to be effective in preventing HFS by compensating for thymidylate deficiency to facilitate the replication and repair of DNA and thus causing the escape from cellular senescence. These data highlight the importance of DNA damage-mediated cellular senescence in the etiology of HFS and provide a potential therapeutic anchor point for fluoropyrimidine-induced HFS.

This paper reports the design, fabrication, and characterization of a MEMS capacitive accelerometer with an asymmetrical comb finger arrangement. By optimizing the ratio of the gaps of a rotor finger to its two adjacent stator fingers, the sensitivity of the accelerometer is maximized for the same comb finger area. With the fingers’ length, width, and depth at 120 μm, 4 μm, and 45 μm, respectively, the optimized finger gap ratio is 2.5. The area of the proof mass is 750 μm × 560 μm, which leads to a theoretical thermomechanical noise of 9 μg/√Hz. The accelerometer has been fabricated using a modified silicon-on-glass (SOG) process, in which a groove is pre-etched into the glass to hold the metal electrode. This SOG process greatly improves the silicon-to-glass bonding yield. The measurement results show that the resonant frequency of the accelerometer is about 2.05 kHz, the noise floor is 28 μg/√Hz, and the nonlinearity is less than 0.5%.

This study presents a novel obstacle evasion method for unmanned aerial vehicle (UAV) clusters in the presence of static and dynamic obstacles. First, a discrete three-dimensional model of the UAV is provided. Second, the proposed improved artificial potential field (APF) is illustrated. In designing the improved scheme, a pseudo-exponential function is fused into the potential field, thus avoiding local extreme points. Frictional resistance is introduced to optimize vibration and maintain stability after reaching the desired endpoints. Meanwhile, the relevant parameters are optimized, and appropriate state limits are defined, thus enhancing the control stability. Third, Lyapunov stability analysis proves that all signals in the closed-loop cluster system are ultimately bounded. Finally, the simulation results demonstrate that the UAV cluster can efficiently reconstruct, form, and maintain formations while avoiding static and dynamical obstacles along with maintaining a safe distance, solving the problem of the local extreme of traditional artificial potential field methods. The proposed scheme is also tested under large-scale multi-UAV scenarios. In conclusion, this study provides valuable insights for engineers working with UAV clusters navigating through formations.

To fill the gaps in the theoretical research on the internal force and deformation of the DSCG, the development law of the internal force and deformation of DSCG was explored in conjunction with the theory of elastic analysis. In addition, a finite element model was established to validate the calculation results. The results showed that using different pre-stressing increment calculation methods affected the calculation results of the composite interface deformation, with the equivalent section method accounting for 0.74% and the principle of the virtual work method for 0.03%. On the other hand, the development of internal forces and deformations in the DSCG was closely related to the magnitude of the load forms and axle weights. At the same time, material non-linearity had less influence on these factors. Finally, the development patterns of the internal forces and deformations of the DSCG with different spans were similar. The specific values were closely related to the span of the guideway, and the interfacial slip, axial force, and deflection of the DSCG with span L = 25 m were 0.60, 0.41, and 0.23 times those of the DSCG with span L = 35 m, respectively. The conclusions of this paper fill the gaps in the theoretical study of multi-system guideways.

Selenium (Se) is an essential trace element known for its significant role in maintaining human health and mitigating disease progression. Selenium and its compounds exhibit high selective cytotoxicity against tumor cells. However, their anti-cervical cancer (CC) effects and underlying mechanisms have not been fully explored. This study found that sodium selenite (SS) inhibits the viability of HeLa and SiHa cells in a dose- and time-dependent manner. Intraperitoneal injection of 3 and 6 mg/kg SS for 14 days in female nude mice significantly inhibited the growth of HeLa cell xenografts without evident hepatotoxicity or nephrotoxicity. RNA sequencing results indicated that the AMP-activated protein kinase (AMPK), Forkhead box protein O (FOXO), and apoptosis signaling pathways are key regulatory pathways in SS’s anti-CC effects, and SS’s inhibition of HeLa cell proliferation may be related to autophagy and ROS-induced apoptosis. Further research has revealed that SS induces cell autophagy and apoptosis through the AMPK/mTOR/FOXO3a pathway, characterized by the upregulation of p-AMPK/AMPK, FOXO3a, LC3-II, cleaved-caspase3, and cleaved-PARP and the downregulation of p-mTOR/mTOR and p62. Additionally, SS impaired mitochondrial function, including decreased mitochondrial membrane potential, mitochondrial Ca2+ overload, and accumulation of mitochondrial reactive oxygen species (mtROS). Pretreatment with Mitoquinone mesylate (Mito Q) and compound C partially reversed SS-induced apoptosis, autophagy, and proliferation inhibition. Pretreatment with 3-methyladenine (3-MA) enhances SS-induced apoptosis and proliferation inhibition in HeLa cells but reverses these effects in SiHa cells. In summary, SS induces apoptosis, autophagy, and proliferation inhibition in HeLa and SiHa cells through the activation of the AMPK/mTOR/FOXO3a signaling pathway via mtROS. Autophagy activation may be a major risk factor for SS-induced apoptosis in SiHa cells but can protect HeLa cells from SS-induced apoptosis. These findings provide new evidence for understanding the molecular mechanisms underlying SS in potential new drug development for CC.