Design of a long-lived Mo2C-MoO2@GC-N electrocatalyst by the ambient DC arc plasma for the hydrogen evolution reaction.

A crucial challenge in hydrogen production through electrolysis is developing inexpensive, earth-abundant, and highly efficient Pt-free electrocatalysts for the hydrogen evolution reaction (HER). Molybdenum carbide is ideal for this application because of its special electrical structure, low cost, and advantageous characteristics. Herein, the long-lived electrocatalysts for HER have been synthesized via the direct current (DC) arc discharge plasma method under ambient air conditions, and the relationship between the properties of materials and catalytic characteristics has been established. The samples differed in the ratio of molybdenum, graphite, and melamine. The sample with the highest proportion of melamine in the initial mixture has Mo2C-MoO2 heterointerfaces, which demonstrates the highest and most stable electrocatalytic activity with the overpotential of 148 mV at 10 mA·cm-2 and Tafel slope of 63 mV·dec-1 in alkaline electrolyte. Meanwhile, the electrodes demonstrated long-lived electrochemical durability for two weeks and investigated the features of forming a stable system for HER.

RAB13 regulates macrophage polarization in sepsis.

To select the core target (RAB13) in sepsis patients' peripheral blood and investigate its molecular functions and possible mechanisms. The peripheral blood of sepsis patients (n = 21) and healthy individuals (n = 9) within 24 h after admission were collected for RNA-seq, and differential gene screening was performed by iDEP online analysis software (P < 0.01; log2FC ≥ 2) and enrichment analysis, the potential core target RAB13 was screened out. The association between RAB13 expression and sepsis severity was explored using multiple datasets in the GEO database, and survival analysis was conducted. Subsequently, peripheral blood mononuclear cells (PBMCs) from sepsis and control groups were isolated, and 10 × single-cell sequencing was used to identify the main RAB13-expressing cell types. Finally, LPS was used to stimulate THP1 cells to construct a sepsis model to explore the function and possible mechanism of RAB13. We found that RAB13 was a potential core target, and RAB13 expression level was positively associated with sepsis severity and negatively correlated with survival based on multiple public datasets. A single-cell sequencing indicated that RAB13 is predominantly localized in monocytes. Cell experiments validated that RAB13 is highly expressed in sepsis, and the knockdown of RAB13 promotes the polarization of macrophages towards the M2 phenotype. This mechanism may be associated with the ECM-receptor interaction signaling pathway. The upregulation of RAB13 in sepsis patients promotes the polarization of M2-like macrophages and correlates positively with the severity of sepsis.

Obstructive sleep apnoea is independently associated with non-alcoholic fatty pancreatic disease.

PURPOSE: This study investigated the prevalence of non-alcoholic fatty pancreatic disease (NAFPD) in patients with obstructive sleep apnoea (OSA) and analysed the correlation between them. MATERIALS AND METHODS: Patients who underwent polysomnography and modified abdominal ultrasound examinations were continuously enrolled and divided into OSA and control groups. The OSA group was further divided into NAFPD and non-NAFPD groups. Differences in various indicators were compared. Binary logistic regression analyses were conducted to identify factors influencing NAFPD. RESULTS: A total of 210 participants were included in the study, including 178 in the OSA group and 32 in the control group. The prevalence of NAFPD, weight, body mass index (BMI), and other indicators were significantly higher in the OSA group than that in the control group (p < 0.05). The above indices, prevalence of hypertension and metabolic associated steatotic liver disease (MASLD), and severity of sleep apnoea were higher in the NAFPD group than that in the non-NAFPD group (p < 0.05). Binary logistic regression analysis showed a 4.4% increased risk of NAFPD for each unit increase in apnea-hypopnea index (AHI) after adjusting for BMI and hypertension. CONCLUSION: The prevalence of NAFPD was higher in the OSA group than that in the control group, and chronic intermittent hypoxia was strongly associated with pancreatic fat infiltration; AHI, BMI, and hypertension were independent risk factors for NAFPD. The NAFPD group had a higher prevalence of obesity and some other diseases.

This study confirms that the modified ultrasound evaluation method can serve as an effective tool for screening pancreatic fat infiltration in OSA patients, aiding in the early identification of the risk of ectopic lipid deposition and metabolic excess.

Effect of uric acid on lipid metabolism assessed via restricted cubic splines: A new insight.

Background: Hyperuricemia can promote both blood lipids and non-alcoholic fatty liver disease (NAFLD). However, the role of the entire uric acid (UA) span, especially low concentrations below hyperuricemia, on lipid metabolism remains unclear. Methods: A cross-sectional study was designed. Data on the age, sex, UA, triglyceride (TG), total cholesterol (TC), high-density lipoprotein (HDL), and low-density lipoprotein (LDL) of 1977 participants, who underwent physical examination, were collected. NAFLD and non-alcoholic fatty pancreas disease (NAFPD) were diagnosed using abdominal ultrasound. Restricted cubic splines (RCS) linear regression model was used to evaluate the effect of the UA span on TG, TC, HDL, and LDL, respectively. RCS logistic regression model was employed to evaluate the effect of the UA span on NAFLD and NAFPD. Results: RCS linear regression model showed that TG was negatively correlated with UA at first, then exhibiting a positive correlation. Meanwhile, HDL was positively correlated with UA at first, then negatively correlated. There was a positive linear correlation between TC and UA (P for nonlinear = 0.578) and a positive nonlinear correlation between LDL and UA (P for nonlinear = 0.021). RCS logistic regression model showed that NAFLD and NAFPD were negatively correlated with UA at first and then positively correlated with UA. Conclusion: our study showed that the entire UA span has a J-shaped effect on some lipids, NAFLD, and NAFPD. Besides, TG and HDL, compared with TC or LDL, may better reflect the status of NAFLD and NAFPD.

The association of sedentary time with sleep disturbances among the US population, 2005 to 2014.

BACKGROUND: Sleep problems increase the risk of premature illness and death. We evaluated the association between sedentary time and sleep disturbances. METHODS: A cross-sectional analysis of the US nationally representative data of 21,414 adults (aged > = 18 years) from National Health and Nutrition Examination Survey (NHANES) (2005-2014) was performed. The data of sleep disturbances were assessed using NHANES questionnaire results, which included the question, "{Have you/has sp} ever been told by a doctor or other health professional that {you have/s/he has} a sleep disorder?". All participants were stratified by quartiles of sedentary behavior distribution, which was the explanatory variable (sedentary time quartile cut points: Q1, 0 < = Q1 < 3 h; Q2, 3 < = Q2 < 5 h; Q3, 5 < = Q3 < 8 h; Q4, 8 < = Q4 < 20 h). We used multivariable logistic regression and the restricted cubic splines (RCS) model to assess the relationship between sedentary time and sleep disturbances. RESULTS: In the unadjusted multivariable logistic regression model (crude model), there was a demonstrated tendency for the odds of sleep disturbances to increase with the sedentary time (Q1 as reference, Q2: OR, 1.31 [95% CI 1.09-1.58] P = 0.005; Q3: OR, 1.62 [95% CI 1.39-1.88] P < 0.001; Q4: OR, 1.75 [95% CI 1.48-2.06] P < 0.001; P for trend < 0.001). In the adjusted model 4, adjustment for gender, age, marital type, education type, race, family poverty index ratio, waist circumference, recreational type, smoke status, drink status, diabetes mellitus status, cardiovascular disease status, sleep duration type, body mass index, the OR in Q2 subgroup didn't significantly increase (Q1 as reference. Q2: OR, 1.18 [95% CI 0.96-1.44] P = 0.1). However, the ORs in Q3 and Q4 (Q3: OR, 1.35 [95% CI 1.14-1.59] P < 0.001; Q4: OR, 1.45 [95% CI 1.21-1.75] P < 0.001) both revealed that the risk of sleep disturbances increased with increasing sedentary time, P for trend < 0.001. The unadjusted RCS model revealed that the risk of sleep disturbances increased non-linearly with increasing sedentary time for total participants (P for non-linearity < 0.001). After adjusting for all covariates, the RCS results revealed that the risk of sleep disturbances increased non-linearly with increasing sedentary time for total participants (P for non-linearity = 0.012). CONCLUSIONS: This study suggested that the longer sedentary time was strongly associated with the sleep disturbances. The protective effect of recreational activities on sleep disturbance, has not been significantly demonstrated.

Leptin A deficiency affecting the mitochondrial dynamics of aged oocytes in medaka (Oryzias latipes).

Mitochondrial dysfunction and metabolic disorder have been associated to age-related subfertility, however, the precise molecular mechanism controlling the development of fertile oocytes in aging females remains elusive. Leptin plays an important role in the maintenance of energy homeostasis, as both excessive or insufficient levels can affect the body weight and fertility of mice. Here, we report that leptin A deficiency affects growth and shortens reproductive lifespan by reducing fertility in medaka (Oryzias latipes). Targeted disruption of lepa (lepa-/-) females reduced their egg laying and fertility compared to normal 3-month-old females (lepa+/+ sexual maturity), with symptoms worsening progressively at the age of 6 months and beyond. Transcriptomic analysis showed that differentially expressed genes involved in metabolic and mitochondrial pathways were significantly altered in lepa-/- ovaries compared with the normal ovaries at over 6 months old. The expression levels of the autophagy-promoting genes ulk1a, atg7 and atg12 were significantly differentiated between normal and lepa-/- ovaries, which were further confirmed by quantitative polymerase chain reaction analysis, indicating abnormal autophagy activation and mitochondrial dysfunction in oocyte development lacking lepa. Transmission electron microscopy observations further confirmed these mitochondrial disorders in lepa-deficient oocytes. In summary, these research findings provide novel insights into how leptin influences female fertility through mitochondrial-mediated oocyte development.

Unveiling the Biodiversity and Conservation Significance of Medog: A Camera-Trapping Survey on Mammals in the Southeastern Tibetan Mountains.

The Medog in southeastern Tibet is home to a diverse range of wild animals. However, research on these mammals' species directories, distribution, and conservation status remains insufficient, despite their crucial role in maintaining ecological balance. The study carried out a camera-trapping survey to assess mammal biodiversity and the significance of mammal protection in their natural habitats in Gedang, Medog. Future directions and application prospects of the study for wildlife conservation in the southeastern Tibetan mountains were also discussed. The survey, spanning from April 2023 to May 2024, with 19,754 camera trap days, revealed 25 mammalian species across five orders and 14 families. Among these, four classified as Endangered, five as Vulnerable, two as Near Threatened on the IUCN Red List, nine were categorized as Critically Endangered or Endangered on the Red List of China's Vertebrates, and seven were China's national first-class key protected wildlife. The order Carnivora exhibited the highest diversity, comprising 12 species. Furthermore, the study filled the knowledge gap regarding the underrepresentation of Gongshan muntjac Muntiacus gongshanensis in IUCN and provided new insights into the recorded coexistence of the Himalayan red panda Ailurus fulgens and Chinese red panda Ailurus styani along the Yarlung Zangbo River for the first time, and also documented new upper elevation limits for four large to medium-sized species. Regarding the relative abundance indices (RAI) captured by camera traps, the most prevalent species identified was the White-cheeked macaque Macaca leucogenys, followed by the Gongshan muntjac and Himalayan serow Capricornis thar. The monitoring also captured a number of domestic dogs and livestock, as well as human disturbances. These findings underscore the importance of conserving these mammals and emphasize the need for conservation efforts to protect their habitats and reduce human activities that threaten their survival, thereby maintaining the ecological balance of the region. Additionally, the research highlighted Gedang's significance to global conservation efforts for mammalian diversity, providing essential data for effective wildlife conservation strategies.

Oncolytic Virus Senecavirus A Inhibits Hepatocellular Carcinoma Proliferation and Growth by Inducing Cell Cycle Arrest and Apoptosis.

Background and Aims: Hepatocellular carcinoma (HCC) is a highly aggressive tumor with limited treatment options and high mortality. Senecavirus A (SVA) has shown potential in selectively targeting tumors while sparing healthy tissues. This study aimed to investigate the effects of SVA on HCC cells in vitro and in vivo and to elucidate its mechanisms of action. Methods: The cell counting kit-8 assay and colony formation assay were conducted to examine cell proliferation. Flow cytometry and nuclear staining were employed to analyze cell cycle distribution and apoptosis occurrence. A subcutaneous tumor xenograft HCC mouse model was created in vivo using HepG2 cells, and Ki67 expression in the tumor tissues was assessed. The terminal deoxynucleotidyl transferase dUTP nick end labeling assay and hematoxylin and eosin staining were employed to evaluate HCC apoptosis and the toxicity of SVA on mouse organs. Results: In vitro, SVA effectively suppressed the growth of tumor cells by inducing apoptosis and cell cycle arrest. However, it did not have a notable effect on normal hepatocytes (MIHA cells). In an in vivo setting, SVA effectively suppressed the growth of HCC in a mouse model. SVA treatment resulted in a significant decrease in Ki67 expression and an increase in apoptosis of tumor cells. No notable histopathological alterations were observed in the organs of mice during SVA administration. Conclusions: SVA inhibits the growth of HCC cells by inducing cell cycle arrest and apoptosis. It does not cause any noticeable toxicity to vital organs.

Nickel-Molybdenum-Based Three-Dimensional Nanoarrays for Oxygen Evolution Reaction in Water Splitting.

Water splitting is an important approach to hydrogen production. But the efficiency of the process is always controlled by the oxygen evolution reaction process. In this study, a three-dimensional nickel-molybdenum binary nanoarray microstructure electrocatalyst is successfully synthesized. It is grown uniformly on Ni foam using a hydrothermal method. Attributed to their unique nanostructure and controllable nature, the Ni-Mo-based nanoarray samples show superior reactivity and durability in oxygen evolution reactions. The series of Ni-Mo-based electrocatalysts presents a competitive overpotential of 296 mV at 10 mA·cm-2 for an OER in 1.0 M KOH, corresponding with a low Tafel slope of 121 mV dec-1. The three-dimensional nanostructure has a large double-layer capacitance and plenty of channels for ion transfer, which demonstrates more active sites and improved charge transmission. This study provides a valuable reference for the development of non-precious catalysts for water splitting.

A synthetic bacterial consortium improved the phytoremediation efficiency of ryegrass on polymetallic contaminated soil.

Polymetallic contamination of soils caused by mining activities seriously threatens soil fertility, biodiversity and human health. Bioremediation is thought to be of low cost and has minimal environmental risk but its effectiveness needs to be improved. This study aimed to identify the combined effect of plant growth and microbial strains with different functions on the enhancement of bioremediation of polymetallic contaminated soil. The microbiological mechanism of bioremediation was explored by amplicon sequencing and gene prediction. Soil was collected from polymetallic mine wastelands and a non-contaminated site for use in a pot experiment. Remediation efficiency of this method was evaluated by planting ryegrass and applying a mixed bacterial consortium comprising P-solubilizing, N-fixing and SO4-reducing bacteria. The plant-microbe joint remediation method significantly enhanced the above-ground biomass of ryegrass and soil nutrient contents, and at the same time reduced the content of heavy metals in the plant shoots and soil. The application of the composite bacterial inoculum significantly affected the structure of soil bacterial communities and increased the bacterial diversity and complexity, and the stability of co-occurrence networks. The relative abundance of the multifunctional genera to which the strains belonged showed a significant positive correlation with the soil nutrient content. Genera related to carbon (C), nitrogen (N), phosphorus (P), and sulphur (S) cycling and heavy metal resistance showed an up-regulation trend in heavy metal-contaminated soils after the application of the mixed bacterial consortium. Also, bacterial strains with specific functions in the mixed consortium regulated the expression of genes involved in soil nutrient cycling, and thus assisted in making the soil self-sustainable after remediation. These results suggested that the remediation of heavy metal-contaminated soil needs to give priority to the use of multifunctional bacterial agents.

16S rRNA, metagenomics and 2bRAD-M sequencing to decode human thanatomicrobiome.

Microorganisms are essential in the decomposition of corpses and play a significant role in forensic science. However, previous studies have primarily focused on animal remains, specifically the gut, skin, and burial environment. Insufficient research has been conducted on the microbiota of human cadavers, especially in cases of advanced decomposition and additional tissues, resulting in a lack of relevant reference data. In this study, the microbiota of eight cadavers at different stages of decomposition were detected using 16S rRNA, metagenomic sequencing and 2bRAD-M sequencing. Nine different sites, including oral and nasal cavities, heart, liver, spleen, lung, kidney, muscle and gut, were analysed and the efficacy of these methods was evaluated. The results showed that 16S rRNA sequencing was the most cost-effective method for the study of cadavers in the early stages of decomposition, whereas for cadaveric tissues in the late stages of decomposition, 2bRAD-M could overcome host contamination more effectively than metagenomic sequencing. This paves the way for new opportunities in data retrieval and promotes in-depth investigations into the microbiota.

Loosely Bounded Exciton with Enhanced Delocalization Capability Boosting Efficiency of Organic Solar Cells.

In organic solar cells (OSCs), electron acceptors have undergone multiple updates, from the initial fullerene derivatives, to the later acceptor-donor-acceptor type non-fullerene acceptors (NFAs), and now to Y-series NFAs, based on which efficiencies have reached over 19%. However, the key property responsible for further improved efficiency from molecular structure design is remained unclear. Herein, the material properties are comprehensively scanned by selecting PC71BM, IT-4F, and L8-BO as the representatives for different development stages of acceptors. For comparison, asymmetric acceptor of BTP-H5 with desired loosely bounded excitons is designed and synthesized. It's identified that the reduction of intrinsically exciton binding energy (Eb) and the enhancement of exciton delocalization capability act as the key roles in boosting the performance. Notably, 100 meV reduction in Eb has been observed from PC71BM to BTP-H5, correspondingly, electron-hole pair distance of BTP-H5 is almost two times over PC71BM. As a result, efficiency is improved from 40% of S-Q limit for PC71BM-based OSC to 60% for BTP-H5-based one, which achieves an efficiency of 19.07%, among the highest values for binary OSCs. This work reveals the confirmed function of exciton delocalization capability quantitatively in pushing the efficiency of OSCs, thus providing an enlightenment for future molecular design.

Portable Copper-Based Electrochemical SERS Sensor for Point-of-Care Testing of Paraquat and Diquat by On-Site Electrostatic Preconcentration.

With the advent of portable Raman spectrometers, the deployment of surface-enhanced Raman spectroscopy (SERS) in point-of-care testing (POCT) has been initiated. Within any analytical framework employing SERS, the acuity and selectivity inherent to the SERS substrate are of paramount importance. In this article, we utilize in situ electrochemical passivation technology to fabricate CuI passivation film, which serves as a flexible copper-based SERS substrate. Furthermore, portable electrochemical SERS (EC-SERS) sensors were prepared by combining this with laser direct writing technology. The detection signal was amplified using electrostatic preconcentration technology, showcasing impressive sensitivity, selectivity, and stability in pesticide detection. The detected concentrations of paraquat and diquat in tea reached as low as 3.36 and 2.43 µg/kg, respectively. Furthermore, the application of electrostatic preconcentration facilitated selective target molecule aggregation on the SERS sensor, markedly increasing Raman signal strength and enabling single-molecule detection. This research introduces an innovative POCT method for pesticides, promising to advance environmental monitoring's analytical capabilities.

Endovascular thrombectomy versus medical management on outcomes with infarct volumes more than 70 mL.

OBJECTIVE: Endovascular thrombectomy (EVT) in patients with large infarct volume remains controversial. The aim of this study is to compare clinical outcomes between EVT and medical management in acute large vessel occlusion with infarct volumes larger than 70 mL on diffusion-weighted magnetic resonance imaging (DWI). METHODS: A prospective observational cohort study was conducted, including patients with anterior cerebral circulation occlusion due to ischemic stroke with infarct volumes larger than 70 mL within 24 h of onset between July 2018 and June 2023. Eligible patients were divided into two groups: the EVT group and the medical management (non-EVT) group. The main outcomes were functional independence and mortality at 90 days. To assess clinical endpoints, we selected variables including age, NIHSS score, infarct volume, and occlusion location for 1:1 propensity score (PS) matching and PS adjustment using inverse probability of treatment weighting (IPTW). RESULTS: Among the 131 identified patients (mean [SD] age, 69.9 [13.7] years; 58 female), the median infarct volume was 123.6 mL. Of these patients, 75 (57.3%) underwent EVT. After PS adjustment, EVT was not associated with functional independence (10.9% vs. 10.9%; p = 1.000) or mortality (43.5% vs. 47.8%; p = 0.675). Additionally, after PS adjustment using IPTW, EVT was also not associated with a functional independence (15.8% vs. 13.7%; p = 0.767) or mortality (46.8% vs. 44.0%; p = 0.762). CONCLUSION: This study provides real-world evidence regarding infarct volumes larger than 70 mL, indicating that EVT does not provide benefits compared to medical management alone when considering age, NIHSS score, infarct volume, and occlusion location.

Comparison of the full-length sequence and sub-regions of 16S rRNA gene for skin microbiome profiling.

The skin microbiome plays a pivotal role in human health by providing protective and functional benefits. Furthermore, its inherent stability and individual specificity present novel forensic applications. These aspects have sparked considerable research enthusiasm among scholars across various fields. However, the selection of specific 16S rRNA hypervariable regions for skin microbiome studies is not standardized and should be validated through extensive research tailored to different research objectives and targeted bacterial taxa. Notably, third-generation sequencing (TGS) technology leverages the full discriminatory power of the 16S gene and enables more detailed and accurate microbial community analyses. Here, we conducted full-length 16S sequencing of 141 skin microbiota samples from multiple human anatomical sites using the PacBio platform. Based on this data, we generated derived 16S sub-region data through an in silico experiment. Comparisons between the 16S full-length and the derived variable region data revealed that the former can provide superior taxonomic resolution. However, even with full 16S gene sequencing, limitations arise in achieving 100% taxonomic resolution at the species level for skin samples. Additionally, the capability to resolve high-abundance bacteria (TOP30) at the genus level remains generally consistent across different 16S variable regions. Furthermore, the V1-V3 region offers a resolution comparable with that of full-length 16S sequences, in comparison to other hypervariable regions studied. In summary, while acknowledging the benefits of full-length 16S gene analysis, we propose the targeting of specific sub-regions as a practical choice for skin microbial research, especially when balancing the accuracy of taxonomic classification with limited sequencing resources, such as the availability of only short-read sequencing or insufficient DNA.IMPORTANCESkin acts as the primary barrier to human health. Considering the different microenvironments, microbial research should be conducted separately for different skin regions. Third-generation sequencing (TGS) technology can make full use of the discriminatory power of the full-length 16S gene. However, 16S sub-regions are widely used, particularly when faced with limited sequencing resources including the availability of only short-read sequencing and insufficient DNA. Comparing the 16S full-length and the derived variable region data from five different human skin sites, we confirmed the superiority of the V1-V3 region in skin microbiota analysis. We propose the targeting of specific sub-regions as a practical choice for microbial research.

Case Report: Isolated Vertigo as the Sole Manifestation of Left Lateral Medullary Infarction Following Contralateral Varicella-Zoster Virus Infection.

Background: Cerebral infarct associated with varicella-zoster virus (VZV) has been reported in the literature, while isolated central dizziness due to lateral medullary infarct (LMI) following VZV infection is rarely reported. Case report: We report the case of a 65-year-old man who presented to the neurology department because of herpes zoster on the right trigeminal nerve distribution. At 12 hours after admission, he developed transient vertigo along with nausea and unsteady walking and left-sided spontaneous horizontal nystagmus, gaze-evoked nystagmus, and upbeat nystagmus. The other usual signs of LMI including Horner syndrome, dysarthria, swallowing difficulty, and hemibody sensory change were absent. Video head impulse indicated decreased head impulse gain of the vestibulo-ocular reflex for the bilateral horizontal, anterior, and posterior semicircular canals with abnormal saccade waves. Suppression head impulse paradigm showed few downward saccades reflecting anti-compensatory saccades after the end of the head impulse back to the head-fixed target and decreased vestibulo-ocular reflex gain values of bilateral semicircular canals. Brain magnetic resonance imaging (MRI) showed a small infarct in the far dorsolateral portion of the left rostral medulla. The cerebrospinal fluid was positive for VZV DNA. Conclusions: In patients with VZV infection who develop dizziness, the possibility of cerebral infarct should be considered. Patients with facial herpes zoster and neurological symptoms always be screened for stroke using MRI and lumbar puncture should be performed and acyclovir administered empirically.

Establishment of a Cleavage-Based Single-Plasmid Dual-Luciferase Surrogate Reporter for the Cleavage Efficiency Evaluation of CRISPR-Cas12a Systems and Its Primary Application.

CRISPR-Cas technology is a widely utilized gene-editing tool that involves gRNA-guided sequence recognition and Cas nuclease-mediated cleavage. The design and evaluation of gRNA are essential for enhancing CRISPR/Cas editing efficiency. Various assays such as single-strand annealing, in vitro cleavage, and T7 endonuclease I (T7EI) are commonly used to assess gRNA-mediated Cas protein cleavage activity. In this study, a firefly luciferase and Renilla luciferase co-expressed and a cleavage-based single-plasmid dual-luciferase surrogate reporter was built to evaluate the gRNA-mediated Cas12a cleavage efficiency. The cleavage activities of CRISPR-Cas12a can be quantitatively determined by the recovery degree of firefly luciferase activity. The cleavage efficiency of CRISPR-Cas12a can be quantitatively measured by the recovery of firefly luciferase activity. By using this system, the cleavage efficiency of CRISPR-Cas12a on hepatitis B virus (HBV)/D expression plasmid was evaluated, revealing a negative correlation between gRNA cleavage efficiency and HBV gene expression measured using an enzyme-linked immunosorbent assay. This simple, efficient, and quantifiable system only requires the dual-luciferase vector and CRISPR-Cas12a vector, making it a valuable tool for selecting effective gRNAs for gene editing.

A novel simplified transperineal prostate biopsy guided by perineal ultrasound.

BACKGROUND: Prostate biopsies are mainly performed through transrectal or perineal approaches, while ultrasound probes are located in the rectum for guidance. However, reports on the use of perineal ultrasound-guided transperineal prostate biopsy (PG-TPPB) are few. METHODS: A retrospective case-control study was designed. A total of 111 patients who underwent PG-TPPB from February 2019 to December 2020 were investigated retrospectively. Simultaneously, 188 patients who underwent transrectal prostate biopsy (TRPB) were included as control. The prostate cancer detection rates (PCDR), complication rates, and application values were compared between the 2 groups. RESULTS: The overall PCDR in the PG-TPPB and TRPB groups were 33.3% (37/111) and 39.9% (75/188), respectively (P = .258). There was no significant difference in the PCDR between the 2 groups under each prostate-specific antigen level (all P > .05). The single-needle PCDR in the PG-TPPB and TRPB groups were 21.5% (277/1 287) and 24.0% (513/2 134), respectively (P = .091). The incidence of complications in the PG-TPPB group was significantly lower than that in the TRPB group (8.1% vs 21.3%, P = .003). CONCLUSIONS: The PCDRs of PG-TPPB and TRPB were the same. However, the postoperative complication rate of PG-TPPB was significantly lower than that of TRPB. Moreover, PG-TPPB required simpler equipment and did not require enema administration, which is suitable for patients with rectal contraindications. ADVANCES IN KNOWLEDGE: The reports on PG-TPPB are few. Our study indicated that PG-TPPB reduced the postoperative complication rate. Moreover, PG-TPPB required simpler equipment. Importantly, PG-TPPB is suitable for patients with rectal contraindications.

Reprogramming of astrocytes and glioma cells into neurons for central nervous system repair and glioblastoma therapy.

Central nervous system (CNS) damage is usually irreversible owing to the limited regenerative capability of neurons. Following CNS injury, astrocytes are reactively activated and are the key cells involved in post-injury repair mechanisms. Consequently, research on the reprogramming of reactive astrocytes into neurons could provide new directions for the restoration of neural function after CNS injury and in the promotion of recovery in various neurodegenerative diseases. This review aims to provide an overview of the means through which reactive astrocytes around lesions can be reprogrammed into neurons, to elucidate the intrinsic connection between the two cell types from a neurogenesis perspective, and to summarize what is known about the neurotranscription factors, small-molecule compounds and MicroRNA that play major roles in astrocyte reprogramming. As the malignant proliferation of astrocytes promotes the development of glioblastoma multiforme (GBM), this review also examines the research advances on and the theoretical basis for the reprogramming of GBM cells into neurons and discusses the advantages of such approaches over traditional treatment modalities. This comprehensive review provides new insights into the field of GBM therapy and theoretical insights into the mechanisms of neurological recovery following neurological injury and in GBM treatment.

Identification of sepsis-associated mitochondrial genes through RNA and single-cell sequencing approaches.

BACKGROUND: Sepsis ranks among the most formidable clinical challenges, characterized by exorbitant treatment costs and substantial demands on healthcare resources. Mitochondrial dysfunction emerges as a pivotal risk factor in the pathogenesis of sepsis, underscoring the imperative to identify mitochondrial-related biomarkers. Such biomarkers are crucial for enhancing the accuracy of sepsis diagnostics and prognostication. METHODS: In this study, adhering to the SEPSIS 3.0 criteria, we collected peripheral blood within 24 h of admission from 20 sepsis patients at the ICU of the Southwest Medical University Affiliated Hospital and 10 healthy volunteers as a control group for RNA-seq. The RNA-seq data were utilized to identify differentially expressed RNAs. Concurrently, mitochondrial-associated genes (MiAGs) were retrieved from the MitoCarta3.0 database. The differentially expressed genes were intersected with MiAGs. The intersected genes were then subjected to GO (Gene Ontology), and KEGG (Kyoto Encyclopedia of Genes and Genomes) analyses and core genes were filtered using the PPI (Protein-Protein Interaction) network. Subsequently, relevant sepsis datasets (GSE65682, GSE28750, GSE54514, GSE67652, GSE69528, GSE95233) were downloaded from the GEO (Gene Expression Omnibus) database to perform bioinformatic validation of these core genes. Survival analysis was conducted to assess the prognostic value of the core genes, while ROC (Receiver Operating Characteristic) curves determined their diagnostic value, and a meta-analysis confirmed the accuracy of the RNA-seq data. Finally, we collected 5 blood samples (2 normal controls (NC); 2 sepsis; 1 SIRS (Systemic Inflammatory Response Syndrome), and used single-cell sequencing to assess the expression levels of the core genes in the different blood cell types. RESULTS: Integrating high-throughput sequencing with bioinformatics, this study identified two mitochondrial genes (COX7B, NDUFA4) closely linked with sepsis prognosis. Survival analysis demonstrated that patients with lower expression levels of COX7B and NDUFA4 exhibited a higher day survival rate over 28 days, inversely correlating with sepsis mortality. ROC curves highlighted the significant sensitivity and specificity of both genes, with AUC values of 0.985 for COX7B and 0.988 for NDUFA4, respectively. Meta-analysis indicated significant overexpression of COX7B and NDUFA4 in the sepsis group in contrast to the normal group (P < 0.01). Additionally, single-cell RNA sequencing revealed predominant expression of these core genes in monocytes-macrophages, T cells, and B cells. CONCLUSION: The mitochondrial-associated genes (MiAGs) COX7B and NDUFA4 are intimately linked with the prognosis of sepsis, offering potential guidance for research into the mechanisms underlying sepsis.