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Abstract rAj-Tspin, a soluble recombinant peptide from Apostichopus japonicus, can inhibit the integrin β1 (ITGB1)/FAK/AKT signaling pathway in hepatocellular carcinoma (HCC) via cell epithelial–mesenchymal transition (EMT) and apoptosis. Zyxin (ZYX) is a focal adhesion protein that is considered a novel mediator of EMT and apoptosis. However, the inhibitory mechanisms of rAj-Tspin in HCC and whether it is related to ZYX are unclear. We examined the antitumor effect of rAj-Tspin on the Huh7 human HCC cell line and on a nude mouse model generated via subcutaneous injection or orthotopic intrahepatic transplantation of Huh7 cells. Our results revealed that rAj-Tspin strikingly reduced the viability and promoted the apoptosis of Huh7 cells and inhibited HCC tumor growth in nude mice. rAj-Tspin inhibited ITGB1 and ZYX protein expression in vivo and in vitro in a dose-dependent manner. Mechanistically, the FAK/AKT signaling pathway and the proliferation and invasion of HCC cells were suppressed upon ITGB1 and ZYX knockdown. Moreover, the effect of ITGB1 overexpression on the growth of HCC cells was inhibited by rAj-Tspin. In contrast, the promoting effect of ITGB1 overexpression could be inhibited by ZYX knockdown. ZYX knockdown had no effect on ITGB1 expression. These findings suggest that ZYX is required for the indispensable role of ITGB1 in rAj-Tspin‐alleviated HCC and provide an important therapeutic target for HCC. In summary, the anti-HCC effect of rAj-Tspin potentially involves the regulation of the ITGB1/ZYX/FAK/AKT pathway, which in turn impacts EMT and apoptosis. Graphical Abstract

Abstract Background Type 2 diabetes mellitus (T2DM) and obstructive sleep apnea (OSA) are mutual risk factors, with both conditions inducing cognitive impairment and anxiety. However, whether OSA exacerbates cognitive impairment and anxiety in patients with T2DM remains unclear. Moreover, TREM2 upregulation has been suggested to play a protective role in attenuating microglia activation and improving synaptic function in T2DM mice. The aim of this study was to explore the regulatory mechanisms of TREM2 and the cognitive and anxiety-like behavioral changes in mice with OSA combined with T2DM. Methods A T2DM with OSA model was developed by treating mice with a 60% kcal high-fat diet (HFD) combined with intermittent hypoxia (IH). Spatial learning memory capacity and anxiety in mice were investigated. Neuronal damage in the brain was determined by the quantity of synapses density, the number and morphology of brain microglia, and pro-inflammatory factors. For mechanism exploration, an in vitro model of T2DM combined with OSA was generated by co-treating microglia with high glucose (HG) and IH. Regulation of TREM2 on IFNAR1-STAT1 pathway was determined by RNA sequencing and qRT-PCR. Results Our results showed that HFD mice exhibited significant cognitive dysfunction and anxiety-like behavior, accompanied by significant synaptic loss. Furthermore, significant activation of brain microglia and enhanced microglial phagocytosis of synapses were observed. Moreover, IH was found to significantly aggravate anxiety in the HFD mice. The mechanism of HG treatment may potentially involve the promotion of TREM2 upregulation, which in turn attenuates the proinflammatory microglia by inhibiting the IFNAR1-STAT1 pathway. Conversely, a significant reduction in TREM2 in IH-co-treated HFD mice and HG-treated microglia resulted in the further activation of the IFNAR1-STAT1 pathway and consequently increased proinflammatory microglial activation. Conclusions HFD upregulated the IFNAR1-STAT1 pathway and induced proinflammatory microglia, leading to synaptic damage and causing anxiety and cognitive deficits. The upregulated TREM2 inT2DM mice brain exerted a negative regulation of the IFNAR1-STAT1 pathway. Mice with T2DM combined with OSA exacerbated anxiety via the downregulation of TREM2, causing heightened IFNAR1-STAT1 pathway activation and consequently increasing proinflammatory microglia. Graphical Abstract

Abstract Background Beta-amyloid (Aβ) deposition in the brain parenchyma is a crucial initiating step in the amyloid cascade hypothesis of Alzheimer’s disease (AD) pathology. Furthermore, dysfunction of plaque-associated microglia, also known as disease-associated microglia (DAM) has been reported to accelerate Aβ deposition and cognitive impairment. Our previous research demonstrated that intermittent hypoxia training (IHT) improved AD pathology by upregulating autophagy in DAM, thereby enhancing oligomeric Aβ (oAβ) clearance. Considering that oAβ internalization is the initial stage of oAβ clearance, this study focused on the IHT mechanism involved in upregulating Aβ uptake by DAM. Methods IHT was administered to 8-month-old APP/PS1 mice or 6-month-old microglial vacuolar protein sorting 35 (VPS35) knockout mice in APP/PS1 background (MG VPS35 KO: APP/PS1) for 28 days. After the IHT, the spatial learning-memory capacity of the mice was assessed. Additionally, AD pathology was determined by estimating the nerve fiber and synapse density, Aβ plaque deposition, and Aβ load in the brain. A model of Aβ-exposed microglia was constructed and treated with IHT to explore the related mechanism. Finally, triggering receptor expressed on myeloid cells 2 (TREM2) intracellular recycling and Aβ internalization were measured using a fluorescence tracing technique. Results Our results showed that IHT ameliorated cognitive function and Aβ pathology. In particular, IHT enhanced Aβ endocytosis by augmenting the intracellular transport function of microglial TREM2, thereby contributing to Aβ clearance. Furthermore, IHT specifically upregulated VPS35 in DAM, the primary cause for the enhanced intracellular recycling of TREM2. IHT lost ameliorative effect on Aβ pathology in MG VPS35 KO: APP/PS1 mice brain. Lastly, the IHT mechanism of VPS35 upregulation in DAM was mediated by the transcriptional regulation of VPS35 by transcription factor EB (TFEB). Conclusion IHT enhances Aβ endocytosis in DAM by upregulating VPS35-dependent TREM2 recycling, thereby facilitating oAβ clearance and mitigation of Aβ pathology. Moreover, the transcriptional regulation of VPS35 by TFEB demonstrates a close link between endocytosis and autophagy in microglia. Our study further elucidates the IHT mechanism in improving AD pathology and provides evidence supporting the potential application of IHT as a complementary therapy for AD. Graphical abstract

Purposes: This study determined the synergy of polymyxin B (POLB) and colistin (COL) with 16 other tested antimicrobial agents in the inhibition of multidrug-resistant Acinetobacter baumannii (MDR-AB). Methods: We used chequerboard assays to determine synergy between the drugs against 50 clinical MDR-AB from a tertiary hospital in the Zhejiang province in 2019, classifying combinations as either antagonistic, independent, additive, or synergistic. The efficacy of hit combinations which showed highest synergistic rate were confirmed using time-kill assays. Results: Both POLB and COL displayed similar bactericidal effects when used in combination with these 16 tested drugs. Antagonism was only observed for a few strains (2%) exposed to a combination of POLB and cefoperazone/sulbactam (CSL). A higher percentage of synergistic combinations with POLB and COL were observed with rifabutin (RFB; 90%/96%), rifampicin (RIF; 60%/78%) and rifapentine (RFP; 56%/76%). Time-kill assays also confirmed the synergistic effect of POLB and rifamycin class combinations. 1/2 MIC rifamycin exposure can achieve bacterial clearance when combined with 1/2 MIC POLB or COL. Conclusion: Nearly no antagonism was observed when combining polymyxins with other drugs by both chequerboard and time-kill assays, suggesting that polymyxins may be effective in combination therapy. The combinations of POLB/COL with RFB, RIF, and RFP displayed neat synergy, with RFB showing the greatest effect.

BackgroundIt has been shown that diabetes is associated with insufficient physical activity among middle-aged and older adults, but the association between different physical activity levels (PAL) and diabetes incidence needs to be further explored.ObjectiveThis study aims to explore the correlation and dose–response relationship between different PAL and the diabetes incidence in middle-aged and older adults.MethodsUtilizing data from the 2018 China Health and Retirement Longitudinal Study (CHARLS), this cross-sectional analysis included 17,226 middle-aged and older adults aged 45 and above. Binary logistic regression models and restricted cubic spline (RCS) were used to explore the correlation and dose–response relationship between different PAL and the incidence of diabetes in the total middle-aged and older adults population as well as in subgroups. Sensitivity analyses were also performed to verify the robustness of the findings.ResultsIn the entire study population, compared with the lowest PAL, participants in the third and fourth quartiles PAL saw diabetes incidence significantly reduced by 16% (p = 0.005) and 33% (p

Background In recent years, Mendelian randomization (MR) has been widely used to infer causality of related disease risk exposures. However, this strategy has not been applied to biliary atresia (BA).Methods Genome-wide association studies (GWAS) data of 41 inflammatory cytokines, 731 immune cell traits, and 1400 metabolites were obtained from public databases as exposure factors. The outcome information was obtained from a GWAS meta-analysis of 499 children with BA and 1928 normal controls. Inverse variance weighting was the primary causality analysis. Cochran Q-test, MR-Egger intercept, MR pleiotropy residual sum and outlier, and ‘leave-one-out’ analyses were used for sensitivity analysis. Reverse MR, MR-Steiger, and Linkage Disequilibrium Score were used to exclude the effects of reverse causality, genetic association, and linkage disequilibrium.Results MR results showed that a total of seven traits had potential causal relationships with BA, including three inflammatory cytokines: eotaxin (odds ratio (OR)=1.45, 95% confidence interval (CI): 1.08 to 1.95, pFDR=0.18), G-CSF (OR=4.21, 95% CI: 1.75 to 10.13, pFDR=0.05) and MCP-1/MCAF (OR=1.53, 95% CI: 1.12 to 2.10, pFDR=0.14); three immune cell traits: CD8dim NKT/T cells ratio (OR=0.59, 95% CI: 0.45 to 0.77, pFDR=0.06), CD8dim NKT counts (OR=0.58, 95% CI: 0.43 to 0.78, pFDR=0.06), CD8dim NKT/lymphocyte ratio (OR=0.63, 95% CI: 0.49 to 0.81, pFDR=0.06); one metabolite: X-12261 levels (OR=2.86, 95% CI: 1.73 to 4.74, pFDR=0.06).Conclusions In this study, eotaxin, G-CSF, MCP-1/MCAF, and X-12261 levels were shown to be risk factors for BA. However, CD8dim NKT/T cells ratio, CD8dim NKT counts, and CD8dim NKT/lymphocyte ratio were protective factors for BA. These findings provided a promising genetic basis for the etiology, diagnosis, and treatment of BA.

Non-dipper blood pressure has been shown to affect cardiovascular outcomes and cognitive function in patients with hypertension. Although some studies have explored the influencing factors of non-dipper blood pressure, there is still relatively little research on constructing a prediction model. This study aimed to develop and validate a simple and practical nomogram prediction model and explore relevant elements that could affect the dipper blood pressure relationship in patients with hypertension. A convenient sampling method was used to select 356 inpatients with hypertension who visited the Affiliated Hospital of Jining Medical College from January 2022 to September 2022. All patients were randomly assigned to the training cohort (75%, n = 267) and the validation cohort (25%, n = 89). Univariate and multivariate logistic regression were utilized to identify influencing factors. The nomogram was developed and evaluated based on the receiver operating characteristic (ROC) curve, the area under the ROC curve (AUC), and decision curve analyses. The optimal cutoff values for the prevalence of dipper blood pressure were estimated. The nomogram was established using six variables, including age, sex, hemoglobin (Hb), estimated glomerular filtration rate (eGFR), ejection fraction (EF), and heart rate. The AUC was 0.860 in the training cohort. The cutoff values for optimally predicting the prevalence of dipper blood pressure were 41.50 years, 151.00 g/L, 117.53 mL/min/1.73 m2, 64.50%, and 75 beats per minute for age, Hb, eGFR, ejection fraction, and heart rate, respectively. In summary, our nomogram can be used as a simple, plausible, affordable, and widely implementable tool to predict the blood pressure pattern of Chinese patients with hypertension.

ABSTRACT The pharmacological activation of peroxisome proliferator‐activated receptor gamma (PPARγ) is a convenient and promising strategy for promoting beige adipocyte biogenesis to combat obesity‐related metabolic disorders. However, thiazolidinediones (TZDs), the full agonists of PPARγ exhibit severe side effects in animal models and in clinical settings. Therefore, the development of efficient and safe PPARγ modulators for the treatment of metabolic diseases is emerging. In this study, using comprehensive methods, we report a previously unidentified ligand‐binding pocket (LBP) in PPARγ and link it to beige adipocyte differentiation. Further virtual screening of 4097 natural compounds based on this novel LBP revealed that saikosaponin A (NJT‐2), a terpenoid compound, can bind to PPARγ to induce coactivator recruitment and effectively activate PPARγ‐mediated transcription of the beige adipocyte program. In a mouse model, NJT‐2 administration efficiently promoted beige adipocyte biogenesis and improved obesity‐associated metabolic dysfunction, with significantly fewer adverse effects than those observed with TZD. Our results not only provide an advanced molecular insight into the structural ligand‐binding details in PPARγ, but also develop a linked selective and safe agonist for obesity treatment.

Manganese (Mn) exposure is a common environmental risk factor for Parkinson's disease (PD), with pathogenic mechanisms associated with dopaminergic neuron damage and neuroinflammation. Mesenchymal stem cells (MSCs)-derived small extracellular vesicles (sEVs) have emerged as a novel therapeutic approach for neural damage repair. The functional sEVs released from MSCs when they are induced into dopaminergic progenitors may have a better repair effect on neural injury. Therefore, we collected sEVs obtained from primary human nasal mucosal mesenchymal stem cells (hnmMSC-sEVs) or cells in the process of dopaminergic progenitor cell differentiation (da-hnmMSC-sEVs), which were cultured in a 3D dynamic system, and observed their repair effects and mechanisms of Mn-induced neural damage by intranasal administration of sEVs. In Mn-exposed mice, sEVs could reach the site of brain injury after intranasal administration, da-hnmMSC enhanced the repair effects of sEVs in neural damage and behavioral competence, as evidenced by restoration of motor dysfunction, enhanced neurogenesis, decreased microglia activation, up-regulation of anti-inflammatory factors, and down-regulation of pro-inflammatory factors. The transcriptomics of hnmMSC-sEVs and da-hnmMSC-sEVs revealed that miRNAs, especially miR-494–3p in sEVs were involved in neuroprotective and anti-inflammatory effects. Overexpression of miR-494–3p in sEVs inhibited Mn-induced inflammation and neural injury, and its repair mechanism might be related to the down-regulation of CMPK2 and NLRP3 in vitro experiments. Thus, intranasal delivery of da-hnmMSC-sEVs is an effective strategy for the treatment of neural injury repair.

Abstract Background Cancer and coronary artery disease (CAD) is reported to often co-exist in same individuals, however, whether cancer is directly associated with anatomical severity of CAD is rarely studied. The present study aimed to observe the relationship between newly diagnosed cancer and anatomical severity of CAD, moreover, to investigate effect of inflammation on the relationship of cancer with CAD. Methods 374 patients with newly diagnosed cancer who underwent coronary angiography (CAG) were enrolled. Through 1:3 propensity score matching (PSM) to cancer patients based on the age and gender among 51,106 non-cancer patients who underwent CAG, 1122 non-cancer patients were selected as control patients. Anatomical severity of CAD was assessed using SYNTAX score (SXscore) based on coronary angiographic image. SXscore ≤ 22 (highest quartile) was defined as SX-low, and SXscore > 22 as SX-high. The ratio of neutrophil to lymphocyte count (NLR) was used to describe inflammation level. Association between cancer and the anatomical severity of CAD was investigated using logistic regression. Results Univariate logistic regression analysis showed a correlation between cancer and anatomical severity of CAD (OR: 1.419, 95% CI: 1.083–1.859; P = 0.011). Cancer was associated with increased risk of SX-high after adjusted for common risk factors of CAD (OR: 1.598, 95% CI: 1.172–2.179, P = 0.003). Significant association between cancer and SX-high was revealed among patients with high inflammation (OR: 1.656, 95% CI: 1.099–2.497, P = 0.016), but not among patients with low inflammation (OR: 1.530, 95% CI: 0.973–2.498, P = 0.089). Conclusions Cancer was associated with severity of CAD, however, the association between the two diseases was significant among patients with high inflammation rather than among patients with low inflammation.

Abstract Background Patient education as an important process of postgraduate nursing education, however in previous studies there was limited study focus on the improvement of nursing students’ patient education in clinical practice.This study examined the effects of a mind mapping based on standardized patient program in the patient education knowledge and communication competence of postgraduate nursing students in clinical setting. Methods The present quasi-experimental study was performed in 2022 on 74 postgraduate nursing students who had taken clinical practice courses at affiliated hospital of Zunyi Medical University. Students were underwent two weeks of mind mapping based on standardized patient program. The outcome measures were patient education knowledge and communication competence evaluated were by the self-designed questionnaire consisting of 6 questions based on the Likert scale and nurse-patient communication competency rating scale respectively, self-efficacy was evaluated by the general self-efficacy scale, and patients’ satisfaction were measured using a self-designed question. Data collection was conducted before and after intervention. Data analysis was performed using SPSS 19.0 software, and descriptive statistics and inferential statistics were performed. Results Significant improvements in patient education knowledge, patient education communication competence, and self-efficacy (all P = 0.000)were observed after intervention. Improvements were also seen in measures of patients’ satisfaction; 12/74 (16.22%) patients reported satisfied at baseline but only 53/74 (71.62%) at the end of intervention. Conclusions A web-based mind maps integrated with standardized patient program could improve patient education knowledge, communication competence,and self-efficacy of postgraduate nursing students in clinical setting.

Abstract Background Salvianolic acid B (Sal B), a water-soluble phenolic compound derived from Salvia miltiorrhiza Bunge, is commonly used in Traditional Chinese Medicine to treat cardiovascular disease. In our previous study, Sal B protected against myocardial fibrosis induced by diabetic cardiomyopathy (DCM). This study aimed to investigate the ameliorative effects and potential mechanisms of Sal B in mitigating myocardial fibrosis induced by DCM. Methods Various methods were used to investigate the effects of Sal B on myocardial fibrosis induced by DCM in vivo and in vitro. These methods included blood glucose measurement, echocardiography, HE staining, Masson’s trichrome staining, Sirius red staining, cell proliferation assessment, determination of hydroxyproline levels, immunohistochemical staining, evaluation of fibrosis-related protein expression (Collagen-I, Collagen-III, TGF-β1, p-Smad3, Smad3, Smad7, and α-smooth muscle actin), analysis of Smad7 gene expression, and analysis of Smad7 ubiquitin modification. Results The animal test results indicated that Sal B significantly improved cardiac function, inhibited collagen deposition and phenotypic transformation, and ameliorated myocardial fibrosis in DCM by upregulating Smad7, thereby inhibiting the TGF-β1 signaling pathway. In addition, cell experiments demonstrated that Sal B significantly inhibited the proliferation, migration, phenotypic transformation, and collagen secretion of cardiac fibroblasts (CFs) induced by high glucose (HG). Sal B significantly decreased the ubiquitination of Smad7 and stabilized the protein expression of Smad7, thereby increasing the protein expression of Smad7 in CFs and inhibiting the TGF-β1 signaling pathway, which may be the potential mechanism by which Sal B mitigates myocardial fibrosis induced by DCM. Conclusion This study revealed that Sal B can improve myocardial fibrosis in DCM by deubiquitinating Smad7, stabilizing the protein expression of Smad7, and blocking the TGF-β1 signaling pathway.

Abstract Objective To evaluate effect of inoculum size of extended-spectrum β-Lactamase (ESBL)-producing-, AmpC-producing-, and KPC-producing Escherichia coli and Klebsiella pneumoniae on the in vitro antibacterial effects of imipenem/relebactam (IMR) and ceftazidime/avibactam (CZA). Methods We compared the impact of inoculum size on IMR and CZA of sixteen clinical isolates and three standard isolates through antimicrobial susceptibility tests, time-kill assays and in vitro PK/PD studies. Results When inoculum size increased from 105 to 107 CFU/mL, an inoculum effect was observed for 26.3% (5/19) and 52.6% (10/19) of IMR and CZA, respectively; time-kill assays revealed that the concentration of CZA increased from ≥ 4 × MIC to 16 × MIC to reach 99.9% killing rate against K. pneumoniae ATCC-BAA 1705 (KPC-2-, OXA-9- and SHV-182-producing) and 60,700 (SHV-27- and DHA-1-producing). While for IMR, a concentration from 1 × MIC to 4 × MIC killed 99.9% of the four strains. When the inoculum size increased to 109 CFU/mL, neither IMR nor CZA showed a detectable antibacterial effect, even at a high concentration. An in vitro PK/PD study revealed a clear bactericidal effect when IMR administered as 1.25 g q6h when inoculum size increased. Conclusion An inoculum effect on CZA was observed more frequent than that on IMR. Among the β-lactamase-producing strains, the inoculum effect was most common for SHV-producing and KPC-producing strains.

Objective: To express the protein HmpA of Salmonella paratyphi A in prokaryotes, and perform bioinformatics analysis on it to provide a theoretical reference for studying the effect of this protein on the nitric oxide (NO) signaling pathway in the host. Methods: The hmpA gene was amplified by PCR and subcloned into the T-vector. Then the expression vector pNdeI-hmpA was constructed and transformed into BL21 (DE3). After induced by IPTG, the recombinant protein expression form was assessed by SDS-PAGE. HmpA was purified using Histrap preload column and identified by Western blot. Bioinformatics technology was used to analyze the characteristics of HmpA. Results: The prokaryotic expression vector pNdeI-hmpA was successfully constructed. HmpA coexisted in the inclusion body and soluble forms under low temperatures after being induced with IPTG. The purified HmpA protein could be detected by the anti-His antibody. Bioinformatics analysis suggested that HmpA was a hydrophilic protein with no transmembrane structural domain or signal peptide. The secondary structure of this protein was mainly composed of α-helix with irregular convolutions, and the tertiary structure model was similar to a ring. The protein structural domain analysis showed that HmpA contains one functional structural domain belonging to the PRK13289 superfamily. There were 32 phosphorylation sites in HmpA. Associated with multiple Salmonella proteases or transcription factors that degrade nitrates and killer NO. Conclusion: This study successfully obtained Salmonella paratyphi A HmpA protein by genetic engineering technology and predicted part of its biological characteristics by bioinformatics methods, which would provide theoretical support for the subsequent uncovering of the influence of HmpA of Salmonella paratyphi A on the nitric oxide signaling pathway in the host.

Introduction: To investigate the diagnostic value of serum MMP-7 and SOX9 levels in patients with biliary atresia (BA) and to further analyze their relationship with the liver fibrosis and prognosis. Methods: Sixty-eight patients hospitalized for jaundice and underwent surgical treatment from March 2019 to February 2022 were retrospectively analyzed, including 44 patients with BA and 24 patients with other cholestatic liver diseases as disease controls (DC). Serum levels of MMP-7 and SOX9 were detected using enzyme-linked immunosorbent assay (ELISA). The diagnostic value of MMP-7 and SOX9 for BA was evaluated using the receiver operating characteristic (ROC) curve. Results: Serum levels of MMP-7 and SOX9 were higher in the BA group compared to the DC group (P = 0.0002, P = 0.0006). The AUC for the combined diagnosis of serum MMP-7, SOX9 and GGT was 0.888 (95 % CI:0.788–0.952, P < 0.0001), which was significantly different from the diagnosis of MMP-7 or SOX9 alone (P = 0.034, P = 0.030). Serum levels of MMP-7 and SOX9 were significantly positively correlated with the liver fibrosis score (rs = 0.41, P = 0.006; rs = 0.58, P < 0.0001). BA patients who maintained native liver survival had lower preoperative serum levels of MMP-7 and SOX9 (P = 0.008, P = 0.0008). Conclusions: The combined diagnosis of BA by MMP-7, SOX9 and GGT has better accuracy than MMP-7 or SOX9 alone. Serum SOX9 levels can assess the severity of BA liver fibrosis and correlate with BA native liver survival, but more sample data are needed to further investigate its predictive value.

Abstract Objectives To validate an appropriate evaluation method of liver fibrosis assessment based on the unique pathological features of biliary atresia (BA) that could well predict its prognosis. Methods A total of 68 patients with BA who underwent Kasai procedure (KP) and an intraoperative liver biopsy, followed up from January 2019 to December 2021, were recruited in a retrospective analysis. Ishak, Metavir, and BA-specific staging systems in relation to outcomes were analyzed using logistic regression, COX proportional hazard regression, Kaplan-Meier analysis, etc. Results Kaplan-Meier analysis determined a significant difference in native liver survival according to the BA-specific stage (p = 0.002). The ROC curve analysis for predicting prognosis showed that the AUC of BA-specific staging combined with iBALF and severe bile duct proliferation (BDP) (0.811, 95% CI: 0.710–0.913, p

Abstract Background Alzheimer's disease (AD) is the most prevalent neurodegenerative disorder. Impaired autophagy in plaque-associated microglia (PAM) has been reported to accelerate amyloid plaque deposition and cognitive impairment in AD pathogenesis. Recent evidence suggests that the transcription factor EB (TFEB)-mediated activation of the autophagy–lysosomal pathway is a promising treatment approach for AD. Moreover, the complementary therapy of intermittent hypoxia therapy (IHT) has been shown to upregulate autophagy and impart beneficial effects in patients with AD. However, the effect of IHT on PAM remains unknown. Methods 8-Month-old APP/PS1 mice were treated with IHT for 28 days. Spatial learning memory capacity and anxiety in mice were investigated. AD pathology was determined by the quantity of nerve fibers and synapses density, numbers of microglia and neurons, Aβ plaque deposition, pro-inflammatory factors, and the content of Aβ in the brain. TFEB-mediated autophagy was determined by western blot and qRT-PCR. Primary microglia were treated with oligomeric Aβ 1–42 (oAβ) combined with IHT for mechanism exploration. Differential genes were screened by RNA-seq. Autophagic degradation process of intracellular oAβ was traced by immunofluorescence. Results In this study, we found that IHT ameliorated cognitive function by attenuating neuronal loss and axonal injury in an AD animal model (APP/PS1 mice) with beta-amyloid (Aβ) pathology. In addition, IHT-mediated neuronal protection was associated with reduced Aβ accumulation and plaque formation. Using an in vitro PAM model, we further confirmed that IHT upregulated autophagy-related proteins, thereby promoting the Aβ autophagic degradation by PAM. Mechanistically, IHT facilitated the nuclear localization of TFEB in PAM, with TFEB activity showing a positive correlation with Aβ degradation by PAM in vivo and in vitro. In addition, IHT-induced TFEB activation was associated with the inhibition of the AKT–MAPK–mTOR pathway. Conclusions These results suggest that IHT alleviates neuronal damage and neuroinflammation via the upregulation of TFEB-dependent Aβ clearance by PAM, leading to improved learning and memory in AD mice. Therefore, IHT may be a promising non-pharmacologic therapy in complementary medicine against AD. Graphical Abstract

As a widely distributed plant in Northeast China, Carex meyeriana Kunth (CMK) is generally considered to have antibacterial properties; however, there is a lack of scientific evidence for this. Therefore, we investigated the chemical composition of CMK extract and its effect against C. albicans. A total of 105 compounds were identified in the alcohol extracts of CMK by UPLC-Q-TOF-MS. Most were flavonoids, with Luteolin being the most represented. Among them, 19 compounds are found in the C. albicans lysates. After treatment with CMK ethanol extract, a significant reduction in the number of C. albicans colonies was observed in a vaginal douche solution from day 5 (p < 0.05). Furthermore, the CMK extract can reduce the number of C. albicans spores. The levels of IL-4, IL-6, IL-10, IL-1β, and TNF-α in vaginal tissues all exhibited a significant decrease (p < 0.05) compared to those in the model group as determined by ELISA. The results of HE staining showed that CMK extract can eliminate vaginal mucosa inflammation. CMK adjusts the vaginal mucosa cells by targeting twenty-six different metabolites and five specific metabolic pathways in order to effectively eliminate inflammation. Simultaneously, the CMK regulates twenty-three types of metabolites and six metabolic pathways against C. albicans infection. So, CMK strongly inhibits the growth of C. albicans and significantly reduces vaginal inflammation, making it a promising candidate for treating C. albicans infection.

With the deteriorating environment and excessive consumption of primary energy, solar energy has become used in buildings worldwide for renewable energy. Due to the fluctuations of solar radiation, a solar photovoltaic (PV) power system is often combined with a storage battery to improve the stability of a building’s energy supply. In addition, the real-time energy consumption pattern of the residual houses fluctuates; a larger size for a PV and battery integrated system can offer more solar energy but also bring a higher equipment cost, and a smaller size for the integrated system may achieve an energy-saving effect. The traditional methods to size a PV and battery integrated system for a detached house are based on the experience method or the traversal algorithm. However, the experience method cannot consider the real-time fluctuating energy demand of a detached house, and the traversal algorithm costs too much computation time. Therefore, this study applies Nondominated Sorting Genetic Algorithm-II (NSGA-II) to size a PV and battery integrated system by optimizing total electricity cost and usage of the grid electricity simultaneously. By setting these two indicators as objectives separately, single-objective genetic algorithms (GAs) are also deployed to find the optimal size specifications of the PV and battery integrated system. The optimal solutions from NSGA-II and single-objective GAs are mutually verified, showing the high accuracy of NSGA-II, and the rapid convergence process demonstrates the time-saving effect of all these deployed genetic algorithms. The robustness of the deployed NSGA-II to various grid electricity prices is also tested, and similar optimal solutions are obtained. Compared with the experience method, the final optimal solution from NSGA-II saves 68.3% of total electricity cost with slightly more grid electricity used. Compared with the traversal algorithm, NSGA-II saves 94% of the computation time and provides more accurate size specifications for the PV and battery integrated system. This study suggests that NSGA-II is suitable for sizing a PV and battery integrated system for a detached house.

Cancer immunotherapies using chimeric antigen receptor (CAR) T cells have tremendous potential and proven clinical efficacy against a number of malignancies. Research and development are emerging to deepen the knowledge of CAR T cell efficacy and extend the therapeutic potential of this novel therapy. To this end, functional characterization of CAR T cells plays a central role in consecutive phases across fundamental research and therapeutic development, with increasing needs for standardization. The functional characterization of CAR T cells is typically achieved by assessing critical effector functions, following co-culture with cell lines expressing the target antigen. However, the use of target cell lines poses several limitations, including alterations in cell fitness, metabolic state or genetic drift due to handling and culturing of the cells, which would increase variabilities and could lead to inconsistent results. Moreover, the use of target cell lines can be work and time intensive, and introduce significant background due to the allogenic responses of T cells. To overcome these limitations, we developed a synthetic bead-based platform (“Artificial Targets”) to characterize CAR T cell function in vitro. These synthetic microparticles could specifically induce CAR T cell activation, as measured by CD69 and CD137 (4-1BB) upregulation. In addition, engagement with Artificial Targets resulted in induction of multiple effector functions of CAR T cells mimicking the response triggered by target cell lines including cytotoxic activity, as assessed by exposure of CD107a (LAMP-1), expression and secretion of cytokines, as well as cell proliferation. Importantly, in contrast to target cells, stimulation with Artificial Targets showed limited unspecific CAR T cell proliferation. Finally, Artificial Targets demonstrated flexibility to engage multiple costimulatory molecules that can synergistically enhance the CAR T cell function and represented a powerful tool for modulating CAR T cell responses. Collectively, our results show that Artificial Targets can specifically activate CAR T cells for essential effector functions that could significantly advance standardization of functional assessment of CAR T cells, from early development to clinical applications.

BackgroundGeriatric diseases (e.g., chronic diseases and geriatric syndromes) may result in impaired physical performance and a decline in the quality of life. The results of previous studies reported the positive effects of comprehensive community-based rehabilitation (CBR) services on physical and social functioning and psychosocial wellbeing. However, to provide adequate and personalised rehabilitation services, it is essential to understand the needs of the older adults population. There have been no studies on the need for CBR in older adults populations that consider their heterogeneity. Therefore, high-quality studies are required to recognise the heterogeneity and latent classes of CBR needs in older adults population groups. This study aims to identify the heterogeneity of the rehabilitation needs of older adults in the community and explore whether older adults with similar characteristics have similar needs through a cross-sectional survey and latent class analysis (LCA) to provide support for personalised rehabilitation services.MethodsThe study is structured into four phases. The first phase will focus on constructing a comprehensive questionnaire to assess rehabilitation needs. In the second phase, a pilot study will be conducted to evaluate the reliability and validity of the completed questionnaire. This step ensures the robustness of the instrument for data collection. The third phase will involve cross-sectional surveys using the finalised questionnaires to collect the necessary data from the targeted population. The fourth phase will focus on conducting LCA to determine the CBR needs of the older adult population.DiscussionThe results of this study will provide novel and critical information for a better understanding of the rehabilitation needs, potential categories, and influencing factors of older adults in the community. The study will be conducted in Guizhou Province in western China, where economic and social development is relatively low, and the results will inform and benefit other regions and developing countries facing similar challenges. However, because of the complete social security and rehabilitation service systems in developed areas, our research results may not fully reflect the situation in these areas. Future studies may need to be conducted in places with different levels of social development.Clinical trial registrationhttps://www.chictr.org.cn/showproj.html?proj=191398, ChiCTR2300071478.

Summary: Nicotinamide adenine dinucleotide (NAD), a nucleotide-containing metabolite, can be incorporated into the RNA 5′-terminus to result in NAD-capped RNA (NAD-RNA). Since NAD has been heightened as one of the most essential metabolites in cells, its linkage to RNA represents a critical but poorly studied modification at the epitranscriptomic level. Here, we design a highly sensitive method, DO-seq, to capture NAD-RNAs. Using Drosophila, we identify thousands of previously unexplored NAD-RNAs and their dynamics in the fly life cycle, from embryo to adult. We show the evidence that chromosomal clustering might be the structural basis by which co-expression can couple with NAD capping on physically and functionally linked genes. Furthermore, we note that NAD capping of cuticle genes inversely correlates with their gene expression. Combined, we propose NAD-RNA epitranscriptome as a hidden layer of regulation that underlies biological processes. DO-seq empowers the identification of NAD-capped RNAs, facilitating functional investigation into this modification.

Summary: Nicotinamide adenine dinucleotide (NAD) can be used as an initiating nucleotide in RNA transcription to produce NAD-capped RNA (NAD-RNA). RNA modification by NAD that links metabolite with expressed transcript is a poorly studied epitranscriptomic modification. Current NAD-RNA profiling methods involve multi-steps of chemo-enzymatic labeling and affinity-based enrichment, thus presenting a critical analytical challenge to remove unwanted variations, particularly batch effects. Here, we propose a computational framework, enONE, to remove unwanted variations. We demonstrate that designed spike-in RNA, together with modular normalization procedures and evaluation metrics, can mitigate technical noise, empowering quantitative and comparative assessment of NAD-RNA across different datasets. Using enONE and a human aging cohort, we reveal age-associated features of NAD-capping and further develop an accurate RNA-based aging clock that combines signatures from both transcriptome and NAD-modified epitranscriptome. enONE facilitates the discovery of NAD-RNA responsive to physiological changes, laying an important foundation for functional investigations into this modification.

Abstract Objective Using latent class to analyze whether there are subtypes of health behaviors in patients with PCOS can be addressed using targeted interventions. Methods October 2021 to June 2022, 471 PCOS patients were surveyed using the Health Promoting Lifestyle Profile Questionnaire. Latent class analysis (LCA) was used to identify subgroups of PCOS patients. Subsequent multinomial latent variable regressions identified factors that were associated with health behaviors. Results A three-class subtypes was the optimum grouping classification: (1)High healthy behavior risk; (2)high healthy responsibility and physical activity risk; (3)low healthy behavior risk. The multinomial logistic regression analysis revealed that (1)Single (OR = 2.061,95% CI = 1.207–3.659), Education level is primary school or below (OR = 4.997,95%CI = 1.732–14.416), participants is student (OR = 0.362,95%=0.138–0.948), participants with pregnancy needs (OR = 1.869,95%=1.009–3.463) were significantly more likely to be in the high healthy behavior risk subtypes; (2)The older the age (OR = 0.953,95%=0.867–1.047) and the larger the WC (OR = 0.954,95%=0.916–0.993), participants is married (OR = 1.126,95%=0.725–1.961), participants is employed ( OR = 1.418,95%=0.667–3.012) were significantly more likely to be in the high health responsibility and physical activity risk subtypes. Conclusion Patients with PCOS are a heterogeneous population with potential subtypes that may be suitable for customized multi-level care and targeted interventions.

In order to clarify the preservation effect of theaflavins-soy protein isolate complex coating on Lentinus edodes, a TFs-SPI complex coating agent was prepared by mixing theaflavins and soy protein isolate as subjects, and its structure was characterized by ultraviolet spectroscopy, Fourier transform infrared spectroscopy, differential scanning calorimetry and X-ray power diffractometry. Meanwhile, the effect of TFs-SPI complex coating on sensory quality, nutrition indicators and total plate count of fresh Lentinus edodes stored at 4 ℃for 20 d were evaluated. The results showed that TFs and SPI could spontaneously form a stable complex through non-covalent interactions. Ultraviolet spectra demonstrated that the interaction between TFs and SPI caused a red shift in the maximum absorption wavelength of SPI and changed the conformation of SPI. Differential scanning calorimetry and X-ray diffraction analysis indicated that TFs-SPI complex was in the amorphous state. Fourier transform infrared spectra revealed that secondary structure of SPI was changed by TFs. Compared with the control, TFs-coated and SPI-coated groups, Lentinus edodes in the TFs-SPI complex coating group showed significant improvement in several qualities during storage, including slowed down the variations of sensory scores and pH values, reduced weight loss and soluble solids content, improved soluble protein and reducing sugar content, as well as inhibited the growth of bacteria. In conclusion, TFs-SPI complex coating treatment could futher maintain the quality of Lentinus edodes, and prolong their shelf life.

Abstract Background Health literacy is a crucial factor that affects health outcomes. Understanding the current status of health literacy among patients with polycystic ovary syndrome (PCOS) is the basis for helping patients better manage risk factors and improve their health outcomes. This study aimed to explore the status of and factors influencing health literacy in patients with PCOS, and to validate the pathway between health literacy, quality of life, and self-efficacy for these patients. Methods A cross-sectional study was conducted using a convenience sample of 300 patients with PCOS in the gynecology outpatient clinic of a tertiary hospital in Zunyi from March to September 2022. Data on health literacy, demographic features, quality of life, and self-efficacy were collected. Multiple stepwise linear regression was conducted to assess the risk factors associated with health literacy for the study participants. A structural equation model was used to construct and validate the pathways. Results Most participants exhibited low health literacy (3.61 ± 0.72), and only 25.70% had adequate health literacy. Multiple regression analysis revealed that the main factors associated with health literacy among participants included Body Mass Index (BMI) (B = −0.95, p

Objective Healthy behaviours are important for people with polycystic ovary syndrome (PCOS). However, adopting and maintaining lifestyle changes involves a complex set of behavioural changes, which most patients fail to adhere to. The current research on health literacy, which includes individual and social skills needed in health self-management, in patients with PCOS is limited. Therefore, this study aimed to explore health literacy, health behaviours and the relationship between the two to expand research on PCOS management.Design Cross-sectional study.Setting A gynaecological outpatient clinic.Participants A total of 286 patients with PCOS (≥18 years, diagnosed via Rotterdam criteria, able to self-report and give informed consent) were recruited from March to June 2022.Outcome measures Health literacy and health behaviours in patients with PCOS were assessed using the Health Literacy Management Scale and the Health Promoting Lifestyle Profile (Chinese version). Multiple logistic regression was used to analyse the factors influencing health literacy, and linear regression to assess the relationship between health literacy and health behaviours, with an F-significance test; p

Background: Cyclovirobuxine D (CVB-D) is a natural alkaloid that exhibits multiple pharmacological activities, such as anti-inflammatory, anti-oxidative stress, and anti-cancer properties. However, its specific protective mechanism of action for myocardial hypertrophy remains unresolved. Purpose: This work was to investigate the ameliorative impact of CVB-D in myocardial hypertrophy, and to elucidate aldosterone (ALD)-induced myocardial hypertrophy by inhibiting the SIRT3 mediated Nrf2 activation. Methods: The myocardial hypertrophy model was reproduced by ALD both in vitro and in vivo, and the protective effect of CVB-D on myocardium and mitochondria was evaluated by TEM, H&E, qPCR, Western blot and ChIP. An immunoprecipitation experiment was adopted to evaluate the acetylation level of Nrf2 and the binding between SIRT3 and Nrf2. Additionally, bardoxolone-methyl (BAR, an Nrf2 agonist), ML385 (an Nrf2 inhibitor), resveratrol (RES, a SIRT3 agonist), and 3-TYP (a SIRT3 inhibitor) were used to confirm the molecular mechanism of CVB-D. Lastly, a molecular docking technique was employed to predict the binding site of SIRT3 and Nrf2 proteins. Results: Our findings suggested that CVB-D improved mitochondrial function, leading to a reduction in ALD-induced cardiomyocyte hypertrophy. By CVB-D treatment, there was an activation of mutual regulation between Nrf2 and SIRT3. Specifically, CVB-D resulted in the increase of Nrf2 protein in the nucleus and activated Nrf2 signaling pathway, thus up-regulating SIRT3. The activation of SIRT3 and the protective action of mitochondrion disappeared because of the intervention of ML385. After CVB-D activated SIRT3, the acetylation level of Nrf2 decreased, followed by activating the Nrf2 pathway. The activation of Nrf2 and mitochondrial protection by CVB-D were reversed by 3-TYP. Our results are also supported by Co-IP and molecular docking analysis, revealing that CVB-D promotes SIRT3-mediated Nrf2 activation. Conclusion: Thus, CVB-D ameliorates ALD-induced myocardial hypertrophy by recovering mitochondrial function by activating the mutual regulation of Nrf2 and SIRT3. Thus, CVB-D could be a beneficial drug for myocardial hypertrophy.

PM2.5 can cause adverse health effects via several pathways, such as inducing pulmonary and systemic inflammation, penetration into circulation, and activation of the autonomic nervous system. In particular, the impact of PM2.5 exposure on the liver, which plays an important role in metabolism and detoxification to maintain internal environment homeostasis, is getting more attention in recent years. In the present study, C57BL/6J mice were randomly assigned and treated with PM2.5 suspension and PBS solution for 8 weeks. Then, hepatic tissue was prepared and identified by metabolomics analysis and transcriptomics analysis. PM2.5 exposure can cause extensive metabolic disturbances, particularly in lipid and amino acids metabolic dysregulation.128 differential expression metabolites (DEMs) and 502 differently expressed genes (DEGs) between the PM2.5 exposure group and control group were detected. The Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses showed that DEGs were significantly enriched in two disease pathways, non-alcoholic fatty liver disease (NAFLD) and type II diabetes mellitus (T2DM), and three signaling pathways, which are TGF-beta signaling, AMPK signaling, and mTOR signaling. Besides, further detection of acylcarnitine levels revealed accumulation in liver tissue, which caused restricted lipid consumption. Furthermore, lipid droplet accumulation in the liver was confirmed by Oil Red O staining, suggesting hepatic steatosis. Moreover, the aberrant expression of three key transcription factors revealed the potential regulatory effects in lipid metabolic disorders, the peroxisomal proliferative agent-activated receptors (PPARs) including PPARα and PPARγ is inhibited, and the activated sterol regulator-binding protein 1 (SREBP1) is overexpressed. Our results provide a novel molecular and genetic basis for a better understanding of the mechanisms of PM2.5 exposure-induced hepatic metabolic diseases, especially in lipid metabolism.

Allergic rhinitis (AR) and adenoid hypertrophy (AH) are common nasal diseases in children. Studies have shown that heavy metals are environmental risk factors for nasal diseases, and the pathogenic mechanisms may be related to dysregulation of nasal mucosal microbiota. However, it is unclear how heavy metal exposure relates to the nasal mucosal microbiota in nasal diseases. Therefore, we explored serum metal exposure levels and nasal mucosal microbiota composition in children with different nasal disease, and further studied the potential correlation between metal exposure and disease-related taxa. There were 64 children recruited for this study. The 23 metals concentrations in serum were measured by inductively coupled plasma mass spectrometry, and nasal mucosal bacteria was identified by 16S rRNA sequencing. Nasal diseases (AR and AH) in children were associated with alterations in the abundance and diversity of the nasal mucosal microbiota. The nasal microbiota of children with AR showed lower diversity, while the microbiota of children with AH showed higher diversity. Linear discriminant analysis Effect Size showed 108 differentially abundant taxa between AR and control groups, 35 differentially abundant taxa among large adenoid, moderate adenoid and small adenoid groups. The serum zinc concentration was negatively correlated with Pielou's eveness index and Simpson's Index in children classified by adenoid size. The spearman correlation analysis showed that multiple disease-related taxa were closely associated with metal concentrations in serum. Our findings may support a link between metal exposure and the diversity and composition of nasal bacteria in children with nasal disease, which present new evidence for the effects of metals on children health.

Unlike traditional user-item recommendation tasks (e.g., movie or consumer-product recommendation), reciprocal recommender systems (RRSs) (e.g., online dating services and job-recruitment sites) must consider the interests of both two users. Pair matching prediction can improve the efficiency with which RRSs match potential partners. Graph Neural Networks (GNNs) are powerful models for learning representations of attributed graphs and information circulation between nodes. GNNs greatly facilitate link prediction in the area of user-item recommender systems but have not been extensively applied to RRS. In this study, we present a novel method for pair matching prediction that learns the reciprocal information circulation between users: not only side information about them but also structural information about their behavior histories. In contrast to earlier RRSs, which focus on response prediction, ours predicts both send and reply signals. Moreover, we introduce negative sample mining to explore the effect of different types of multiple samples on recommendation accuracy in real applications. Testing our method on data provided by an online dating service, we achieved an AUC of 73.15% (an absolute improvement of over 3.20% point above baseline) and an AP of 26.01% (an absolute improvement of over 2.79%) on send prediction; an AUC of 68.95% (an absolute improvement of over 1.74%) and an AP of 23.02% (an absolute improvement of over 0.70%) on reply prediction; an AUC of 71.26% (over 4.35% point absolute improvement) and an AP of 23.95% (over 0.30% point absolute improvement) on fusion reciprocal prediction.

Abstract Backround The supraclavicular plexus block (SCB) and interscalene plexus block (ISB) have the potential to pulmonary function, the duration of the potential remains uncertain. So, we compared the effect of SCB and ISB on pulmonary function, especially the duration time. Methods Ninety-six patients were finally allocated to group I and group S. The ISB and the SCB procedures were performed with ultrasound guidance before anesthesia induction. An investigator recorded the diaphragm mobility and respiratory function test indicators before the block (T0) and at 30 min (T30 min), 4 h (T4), 8 h (T8), and 12 h (T12) after the block. The diaphragmatic paralysis rate was calculated for above timepoint. The VAS, the recovery time for the sensory and motor block, and adverse reactions within 24 h of administering the block were also recorded. Results The recovery times of diaphragm mobility in group I were longer than those in group S. Compared with group I, group S had a significantly lower diaphragmatic paralysis rate during eupnea breathing at T30 min and T8 after the block. Similarly, group S had a significantly lower diaphragmatic paralysis rate at deep breathing at T30 min, T8, and T12 after the block. The recovery times of FEV1 and FVC in group I were longer than those in group S. The other results were not statistically significant. Conclusions Ultrasound-guided ISB resulted in a longer periods with a suppressive effect on pulmonary function than SCB. Trials registration 17/12/2019, ChiCTR1900028286.

AbstractPak choi (Brassica rapa ssp. chinensis Makino) is a typical seed vernalization vegetable. Because premature bolting in spring causes significant economic losses, it is very important to clarify the flower formation mechanism to prevent this. Cytokinins are important plant hormones involved in regulating plant growth and development. To understand the relationship between cytokinin metabolism and flowering of pak choi, in this study, we determined the cytokinin trans-zeatin content in shoot apices of pak choi at different developmental stages by enzyme-linked immunosorbent assay. The results showed that cytokinin levels increased significantly after low temperature treatment at 4 °C, and continued to increase with vegetative growth after transplanting, reaching a peak at the critical period which is immediately prior to flower buds’ differentiation (S0), then decreased thereafter. The levels for low temperature treatment were consistently higher than controls. To explore the molecular mechanism underpinning the cytokinin changes, expression of homologous genes encoding cytokinin metabolic enzymes was analysed by transcriptome sequencing. The expression levels of nine genes (Bra004037, Bra023701, Bra002204, Bra014968, Bra028326, Bra028182, Bra034022, Bra009143 and Bra005869) were consistent with the changes in cytokinin content. The correlation between differentially expressed genes and cytokinin content in different developmental stages of apexes was analysed, and six closely related genes (Bra023701, Bra002204, Bra014968, Bra028182, Bra005869 and Bra009143) were identified. The results help to illuminate the molecular mechanisms controlling flowering of pak choi.

With increasing consumption of primary energy and deterioration of the global environment, clean energy sources with large reserves, such as natural gas, have gradually gained a higher proportion of the global energy consumption structure. Monitoring and predicting consumption data play a crucial role in reducing energy waste and improving energy supply efficiency. However, owing to factors such as high monitoring device costs, safety risks associated with device installation, and low efficiency of manual meter reading, monitoring natural gas consumption data at the household level is challenging. Moreover, there is a lack of methods for predicting natural gas consumption at the household level in residential areas, which hinders the provision of accurate services to households and gas companies. Therefore, this study proposes a gas consumption monitoring method based on the K-nearest neighbours (KNN) algorithm. Using households in a residential area in Xi’an as research subjects, the feasibility of this monitoring method was validated, achieving a model recognition accuracy of 100%, indicating the applicability of the KNN algorithm for monitoring natural gas consumption data. In addition, this study proposes a framework for a natural gas consumption prediction system based on a backpropagation (BP) neural network.

ABSTRACTST11 is the most common lineage among carbapenem-resistant Klebsiella pneumoniae (CRKP) infections in Asia. Diverse morphotypes resulting from genetic mutations are associated with significant differences in microbial characteristics among K. pneumoniae isolates. Here, we investigated the genetic determinants and critical characteristics associated with distinct morphotypes of ST11 CRKP. An ST11-KL47 CRKP isolate carrying a pLVPK-like virulence plasmid was isolated from a patient with a bloodstream infection; the isolate had the “mcsw” morphotype. Two distinct morphotypes (“ntrd” and “msdw”) were derived from this strain during in vitro passage. Whole genome sequencing was used to identify mutations that cause the distinct morphotypes of ST11 CRKP. Transmission electron microscopy, antimicrobial susceptibility tests, growth assays, biofilm formation, virulence assays, membrane permeability assays, and RNA-seq analysis were used to investigate the specific characteristics associated with different morphotypes of ST11 CRKP. Compared with the parental mcsw morphotype, the ntrd morphotype resulted from mutation of genes involved in capsular polysaccharide biosynthesis (wza, wzc, and wbaP), a result validated by gene knockout experiments. This morphotype showed capsule deficiency and lower virulence potential, but higher biofilm production. By contrast, the msdw morphotype displayed competition deficiency and increased susceptibility to chlorhexidine and polymyxin B. Further analyses indicated that these characteristics were caused by interruption of the sigma factor gene rpoN by insertion mutations and deletion of the rpoN gene, which attenuated membrane integrity presumably by downregulating the phage shock protein operon. These data expand current understanding of genetic, virulence, and antimicrobial resistance characteristics associated with distinct morphotypes in ST11 CRKP.

Abstract Background High-altitude cerebral edema (HACE) is a serious and potentially fatal brain injury that is caused by acute hypobaric hypoxia (HH) exposure. Vasogenic edema is the main pathological factor of this condition. Hypoxia-induced disruptions of tight junctions in the endothelium trigger blood‒brain barrier (BBB) damage and induce vasogenic edema. Nuclear respiratory factor 1 (NRF1) acts as a major regulator of hypoxia-induced endothelial cell injury, and caveolin-1 (CAV-1) is upregulated as its downstream gene in hypoxic endothelial cells. This study aimed to investigate whether CAV-1 is involved in HACE progression and the underlying mechanism. Methods C57BL/6 mice were exposed to HH (7600 m above sea level) for 24 h, and BBB injury was assessed by brain water content, Evans blue staining and FITC-dextran leakage. Immunofluorescence, transmission electron microscope, transendothelial electrical resistance (TEER), transcytosis assays, and western blotting were performed to confirm the role and underlying mechanism of CAV-1 in the disruption of tight junctions and BBB permeability. Mice or bEnd.3 cells were pretreated with MβCD, a specific blocker of CAV-1, and the effect of CAV-1 on claudin-5 internalization under hypoxic conditions was detected by immunofluorescence, western blotting, and TEER. The expression of NRF1 was knocked down, and the regulation of CAV-1 by NRF1 under hypoxic conditions was examined by qPCR, western blotting, and immunofluorescence. Results The BBB was severely damaged and was accompanied by a significant loss of vascular tight junction proteins in HACE mice. CAV-1 was significantly upregulated in endothelial cells, and claudin-5 explicitly colocalized with CAV-1. During the in vitro experiments, hypoxia increased cell permeability, CAV-1 expression, and claudin-5 internalization and downregulated tight junction proteins. Simultaneously, hypoxia induced the upregulation of CAV-1 by activating NRF1. Blocking CAV-1-mediated intracellular transport improved the integrity of TJs in hypoxic endothelial cells and effectively inhibited the increase in BBB permeability and brain water content in HH animals. Conclusions Hypoxia upregulated CAV-1 transcription via the activation of NRF1 in endothelial cells, thus inducing the internalization and autophagic degradation of claudin-5. These effects lead to the destruction of the BBB and trigger HACE. Therefore, CAV-1 may be a potential therapeutic target for HACE. Video abstract

The immune cell constituents and localisation within human hepatocellular carcinoma is not fully understood. Here the authors use single cell RNA sequencing of HCC from four different tissue sites and show differences between primary and metastatic tumours, tumour associated macrophages and immune cell populations.

Abstract This paper proposes a dual‐stator switched reluctance generator (DSSRG) which can be applied to underwater power generation. DSSRG is connected with the propeller. When the propeller deflects, it can drive the generator to deflect for underwater power generation and improve the conversion efficiency of the motor. Firstly, taking the underwater propeller as the research object, the pressure and torque characteristics of the propeller at different deflection angles are studied by using CFX software. Then, the structure of DSSRG is modelled and analysed, and the voltage and current characteristics of DSSRG at different speeds and deflection angles are analysed. Finally, an underwater generator prototype is established to verify the rationality of the underwater DSSRG and the feasibility and stability of the power generation system.

The Sanmenxia Basin, located on the southeastern margin of the Shanxi rift and filled with Cretaceous-Paleogene fluvial and lacustrine sediments, is a faulted basin bounded by a series of normal strike-slip faults. This provides a valuable opportunity to investigate the early tectonic deformation in the Fenwei graben. In this study, we report an integrated rock magnetism and AMS analysis of two sections from the Sanmenxia Basin spanning an interval from the Late Cretaceous to the Eocene. The rock magnetic results demonstrate that magnetite and hematite are the main magnetic carriers of remanence, and paramagnetic minerals and hematite are major contributors to the AMS in both sections of the Sanmenxia Basin. Together with the relatively low-corrected anisotropy values, the tightly grouped minimum principal axes are almost perpendicular to the bedding plane, and the well-defined magnetic lineation is generally parallel to the bedding trend, indicating that the primary sedimentary fabrics in the Sanmenxia Basin were overprinted by the initial deformation. The NW–SE magnetic lineation denotes the NW–SE stretching during the Late Cretaceous-Eocene. The stretching process may have been controlled by the Indian-Eurasian convergence and/or the subduction of the western Pacific plate.

IntroductionHypoxia-induced neuronal damage is the primary cause of cognitive impairment induced by high-altitude exposure. Microglia play a crucial regulatory role in the central nervous system (CNS) homeostasis and synaptic plasticity. M1-type polarized microglia are suspected to be responsible for CNS injury under hypoxic conditions, but the exact molecular mechanism is still unelucidated.MethodsCX3CR1 knock out and wide type mice were exposed to a simulated plateau at 7000 m for 48 h to construct the model of hypobaric hypoxia-induced memory impairment. The memory impairment of mice was assessed by Morris water maze. The dendritic spine density in the hippocampus was examined by Golgi staining. The synapses in the CA1 region and the number of neurons in the DG region were examined by immunofluorescence staining. The synapses in microglia activation and phagocytosis were examined by immunofluorescence. The levels of CX3CL1/CX3CR1 and their downstream proteins were detected. CX3CR1 knockout primary microglia were treated with CX3CL1 combined with 1% O2. The levels of proteins related to microglial polarization, the uptake of synaptosome and phagocytotic ability of microglia were detected.ResultsIn this study, mice exposed to a simulated 7000 m altitude for 48 h developed significant amnesia for recent memories, but no significant change in their anxiety levels was observed. Hypobaric hypoxia exposure (7000 m altitude above sea level for 48 h) resulted in synapse loss in the CA1 region of the hippocampus, but no significant changes occurred in the total number of neurons. Meanwhile, microglia activation, increased phagocytosis of synapses by microglia, and CX3CL1/CX3CR1 signal activation were observed under hypobaric hypoxic exposure. Further, we found that after hypobaric hypoxia exposure, CX3CR1-deficient mice showed less amnesia, less synaptic loss in the CA1 region, and less increase in M1 microglia, compared to their wildtype siblings. CX3CR1-deficient microglia did not exhibit M1-type polarization in response to either hypoxia or CX3CL1 induction. Both hypoxia and CX3CL1 induced the phagocytosis of synapses by microglia through the upregulation of microglial phagocytosis.DiscussionThe current study demonstrates that CX3CL1/CX3CR1 signal mediates the M1-type polarization of microglia under high-altitude exposure and upregulates microglial phagocytosis, which increases the phagocytosis of synapses in the CA1 region of the hippocampus, causing synaptic loss and inducing forgetting.

Klebsiella pneumoniae can cause widespread infections and is an important factor of hospital- and community-acquired pneumonia. The emergence of hypervirulent K. pneumoniae poses a serious clinical therapeutic challenge and is associated with a high mortality. The goal of this work was to investigate the influence of K. pneumoniae infection on host cells, particularly pyroptosis, apoptosis, and autophagy in the context of host–pathogen interactions to better understand the pathogenic mechanism of K. pneumoniae. Two clinical K. pneumoniae isolates, one classical K. pneumoniae isolate and one hypervirulent K. pneumoniae isolate, were used to infect RAW264.7 cells to establish an in vitro infection model. We first examined the phagocytosis of macrophages infected with K. pneumoniae. Lactate dehydrogenase (LDH) release test, and calcein-AM/PI double staining was conducted to determine the viability of macrophages. The inflammatory response was evaluated by measuring the pro-inflammatory cytokines and reactive oxygen species (ROS) production. The occurrence of pyroptosis, apoptosis, and autophagy was assessed by detecting the mRNA and protein levels of the corresponding biochemical markers. In addition, mouse pneumonia models were constructed by intratracheal instillation of K. pneumoniae for in vivo validation experiments. As for results, hypervirulent K. pneumoniae was much more resistant to macrophage-mediated phagocytosis but caused more severe cellular damage and lung tissues damage compared with classical K. pneumoniae. Moreover, we found increased expression of NLRP3, ASC, caspase-1, and GSDMD associated with pyroptosis in macrophages and lung tissues, and the levels were much higher following hypervirulent K. pneumoniae challenge. Both strains induced apoptosis in vitro and in vivo; the higher apoptosis proportion was observed in infection caused by hypervirulent K. pneumoniae. Furthermore, classical K. pneumoniae strongly triggered autophagy, while hypervirulent K. pneumoniae weakly activated this process. These findings provide novel insights into the pathogenesis of K. pneumoniae and may form the foundation for the future design of treatments for K. pneumoniae infection.

BackgroundUnderstanding the relationship between anxiety, depression and health-related quality of life (HRQOL) provides important clues to alleviate anxiety, depression and improve HRQOL in patients after severe acute pancreatitis (SAP). The aim of this study was to examine the effects of anxiety and depression on HRQOL in post-SAP patients using structural equation modeling.MethodsA cross-sectional study design was used and 134 patients with SAP from the Affiliated Hospital of Zunyi Medical University were recruited. Data collected included demographic and clinical characteristics, the English Standard Short Form 36 (SF-36) Health Survey, The Self-rating Anxiety Scale (SAS) and The Self-rating Depression Scale (SDS). Structural equation modeling analysis was conducted using the AMOS 24.0 program.ResultsThe mean of HRQOL score was 49.42 (SD = 23.01). The prevalence of anxiety and depression in post-SAP patients was 33.6 and 34.3%, respectively. Both anxiety and depression have a direct negative impact on HRQOL (β = −0.360, p < 0.001; β = −0.202, p = 0.034). Anxiety also negatively affects HRQOL indirectly through depression (β = −0.118, p = 0.043). The analysis of the covariance structure revealed that the resulting model had a reasonable goodness of fit.ConclusionAnxiety and depression reduce the quality of life of SAP patients during recovery. Regular assessment and management of the anxiety and depression status of SAP patients is necessary and will help them improve their HRQOL more effectively.

Introduction: Polygoni Multiflori Radix (PMR) is a type of Chinese herbal medicine with rich chemical composition and pharmacological activity used widely in medicine and food. However, in recent years, there have been increasing numbers of negative reports about its hepatotoxicity. Identification of its chemical constituents for quality control and safe use is very important.Methods: Three solvents of different polarities (water, 70% ethanol, and 95% ethanol solution) were used to extract the compounds from PMR. Extracts were analyzed and characterized by ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-Q-ToF MS/MS) in the negative-ion mode.Results: 152 compounds were detected and identified: 50 anthraquinones, 33 stilbene derivatives, 21 flavonoids, seven naphthalene compounds, and 41 other compounds. Eight other compounds were reported for the first time in the PMR-related literature, and eight other compounds were potentially new compounds.Discussion: This study lays a solid foundation for the screening of toxicity and quality-control indicators of PMR.

Manganese (Mn) accumulates in the central nervous system and can cause neurotoxicity, but the mechanisms of Mn-induced neurotoxicity remain unclear. We performed single-cell RNA sequencing (scRNA-seq) of zebrafish brain after Mn exposure and identified 10 cell types by marker genes: cholinergic neurons, dopaminergic (DA) neurons, glutaminergic neurons, GABAergic neurons, neuronal precursors, other neurons, microglia, oligodendrocyte, radial glia, and undefined cells. Each cell type has its distinct transcriptome profile. Pseudotime analysis revealed that DA neurons had a critical role in Mn-induced neurological damage. Combined with metabolomic data, chronic Mn exposure significantly impaired amino acid and lipid metabolic processes in the brain. Furthermore, we found that Mn exposure disrupted the ferroptosis signaling pathway in the DA neurons in zebrafish. Overall, our study employed joint analysis of multi-omics and revealed ferroptosis signaling pathway is a novel potential mechanism of Mn neurotoxicity.

The MADS-box family, a substantial group of plant transcription factors, crucially regulates plant growth and development. Although the functions of AGL12-like subgroups have been elucidated in Arabidopsis, rice, and walnut, their roles in grapes remain unexplored. In this study, we isolated VvAGL12, a member of the grape MADS-box group, and investigated its impact on plant growth and biomass production. VvAGL12 was found to localize in the nucleus and exhibit expression in both vegetative and reproductive organs. We introduced VvAGL12 into Arabidopsis thaliana ecotype Columbia-0 and an agl12 mutant. The resulting phenotypes in the agl12 mutant, complementary line, and overexpressed line underscored VvAGL12’s ability to promote early flowering, augment plant growth, and enhance production. This was evident from the improved fresh weight, root length, plant height, and seed production, as well as the reduced flowering time. Subsequent transcriptome analysis revealed significant alterations in the expression of genes associated with cell-wall modification and flowering in the transgenic plants. In summary, the findings highlight VvAGL12′s pivotal role in the regulation of flowering timing, overall plant growth, and development. This study offers valuable insights, serving as a reference for understanding the influence of the VvAGL12 gene in other plant species and addressing yield-related challenges.

Fluopicolide is a new benzamide fungicide with a unique mechanism of action and is toxic to some non-target organisms. However, there is a lack of research on the chronic toxicity of fluopicolide to earthworms. In this study, in order to evaluate the chronic toxicity of fluopicolide to earthworms, the levels of reactive oxygen species (ROS) and malondialdehyde (MDA), the activities of superoxide dismutase (SOD), catalase (CAT), and glutathione S-transferase (GST), and DNA oxidative damage (8-hyoxy-2-deoxyguanosine content) in earthworms were measured at 7, 14, 21, and 28 days after exposure to different concentrations (0, 0.1, 0.5, 1, 2.5, 5, and 10 mg/kg) of fluopicolide. In most treatment groups, the ROS levels increased significantly 7 days after exposure and then decreased gradually with an increase in exposure time, a certain dose–effect relationship. The antioxidant enzymes’ activities (SOD and CAT) in most treatment groups were activated, showing an increasing trend at first and then a decreasing trend; however, the CAT activity in the high-concentration treatment group was inhibited 21 days after exposure. The GST activity and MDA content showed an increasing trend at first and then a decreasing trend, which was dependent on the dose. As a biomarker of DNA damage, the 8-OHdG content was positively correlated with the concentration of fluopicolide. The results showed that a low dose of fluopicolide could cause oxidative stress and DNA damage in earthworms.

Accumulating evidence has revealed that the dysregulation of circular RNAs (circRNAs) plays crucial roles in the occurrence and progression of cancers. However, the aberrant expression profile and dysfunction of circRNAs in non-small cell lung cancer (NSCLC) have not been fully explored. Herein, we discovered that a circRNA, hsa_circ_0001666 (circ0001666), was highly expressed in NSCLC tissues and cell lines, and it was positively correlated with NSCLC tumor pathological grade and lymph node metastasis. Moreover, Kaplan-Meier survival analysis implied that NSCLC patients with high circ0001666 expression were negatively correlated with favorable survival. Functionally, circ0001666 could promote migration and invasion of NSCLC cells in vitro and in vivo. Mechanistically, circ0001666 could act as a sponge to miR-1184/miR-548I and upregulate the expression of AGO1, thereby promoting the activation of the phosphatidylinositol 3-kinase (PI3K)/AKT/mTOR signaling pathway in NSCLC cells. Collectively, these findings demonstrated that circ0001666 could serve as an oncogene to promote the migration and invasion of NSCLC via a novel miR-1184/miR-548I/AGO1 axis, which might be a promising therapeutic target for NSCLC treatment.

Background: /Objective: The treatment of bone defect has always been a difficult problem in orthopedic clinic. The search for alternative biodegradable implants is a hot topic. The development of biodegradable magnesium scaffolds for the treatment of bone defects has long been a goal of the public. Methods: In this study, we proposed a porous magnesium scaffold prepared by 3D gel printing and surface modification with an additional calcium phosphate coating and use of its strength, degradability and slow degradation rate in a bone graft substitute material. The porous magnesium granular scaffold was prepared by 3D gel printing technology and modified by DCPD (Dibasic Calcium Phosphate Dihydrate) coating. The biocompatibility, degradation rate, and osteogenic ability of the scaffold were evaluated in vitro and in vivo. Results: The biocompatibility, in vivo degradation and bone defect healing response of the implants were investigated. Porous magnesium scaffolds were successfully prepared, and the strength of sintered scaffolds reached 5.38 ​MPa. The degradation rates of scaffolds were significantly reduced after coating with DCPD. The cell compatibility evaluation showed that DCPD-coated Mg scaffold was suitable for cell proliferation. In vivo biosafety monitoring showed that scaffold implantation did not cause an increase in Mg ion concentration in vivo, and no toxic damage was detected in the liver or kidney. Micro-CT and pathological results showed that a large amount of new bone was formed at 6 weeks. At 12 weeks, approximately 52% of the scaffold volume remained. At 24 weeks, osteogenesis, which was stimulated by some residual scaffold, still can be observed. In summary, this study suggests that 3D gel-printed DCPD-coated porous magnesium scaffolds have great potential as bone graft alternatives. Conclusion: In summary, this study suggests that 3D gel-printed DCPD-coated porous magnesium scaffolds have great potential as bone graft alternatives. The Translational potential of this article: The translational potential of this article is to make use of the advantages of 3D gel printing technology with higher efficiency and lower cost compared with SLM and SLS technologies, and use pure magnesium powder as raw material to prepare degradable porous magnesium metal scaffolds, opening up a new technical route for the preparation of degradable porous magnesium scaffolds which are made for bone defect regeneration in the future.

ABSTRACT: Avian hepatitis E virus (avian HEV) increases poultry mortality and decreases egg production, leading to huge economic losses worldwide. However, there is no effective serological test for avian HEV. Researchers previously created a testing platform using the nanobody (Nb)-horseradish peroxidase (HRP) fusion protein as an ultrasensitive probe to develop competitive ELISA (cELISA) to detect antibodies against different animal viruses. In this study, a rapid and reliable cELISA was developed to test for antibodies against avian HEV using the same platform. Six anti-avian HEV capsid protein nanobodies were selected from an immunized Bactrian camel using phage display technology. The avian HEV-Nb49-HRP fusion protein was expressed and used as a probe for developing a cELISA assay to test for avian HEV antibodies. The cut-off value of the developed cELISA was 22.0%. There was no cross-reaction with other anti-avian virus antibodies, suggesting that the cELISA had good specificity. The coefficients of variation were 0.91% to 4.21% (intra-assay) and 1.52% to 6.35% (inter-assay). Both cELISA and indirect ELISA showed a consistency of 86.7% (kappa = 0.738) for clinical chicken serum samples, and coincidence between cELISA and Western blot was 96.0% (kappa = 0.919). The epitope recognized by Nb49 was located in aa 593-604 of the avian HEV capsid protein, and the peptide (TFPS) in aa 601-604 was essential for binding. The novel cELISA is a saving cost, rapid, useful, and reliable assay for the serological investigation of avian HEV. More importantly, the peptide TFPS may be crucial to immunodominant antigen composition and protection.

The deterioration of environment, the aggravation of energy crisis and the exhaustion of non renewable energy promote the development of green new energy. Among them, wind power generation has become a research hotspot in the field of new energy at home and abroad. In this paper, the deflection double stator switched reluctance generator (DDSSRG) is taken as the research object, the basic modelling and analysis of DDSSRG are carried out, and the magnetic field of the generator is analyzed by using the finite element software. The structure of the generator is optimized by using the central composite design method (CCD). Finally, the experimental platform of the generator is built to verify the feasibility of the theory in structure.