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Abstract Background Lung cancer is one of the most common cancers and causes of cancer death in Canada. Some previous literature suggests that socioeconomic inequalities in lung cancer screening, treatment and survival may exist. The objective of this study was to compare overall survival for immigrants versus long-term residents of Ontario, Canada among patients diagnosed with lung cancer. Methods This population-based retrospective cohort study utilized linked health administrative databases and identified all individuals (immigrants and long-term residents) aged 40 + years diagnosed with incident lung cancer between April 1, 2012 and March 31, 2017. The primary outcome was 5-year overall survival with December 31, 2019 as the end of the follow-up period. We implemented adjusted Cox proportional hazards models stratified by age at diagnosis, sex, and cancer stage at diagnosis to examine survival. Results Thirty-eight thousand seven hundred eighty-eight individuals diagnosed with lung cancer were included in our cohort including 7% who were immigrants. Immigrants were younger at diagnosis and were more likely to reside in the lowest neighbourhood income quintile (30.6% versus 24.5%) than long-term residents. After adjusting for age at diagnosis, neighbourhood income quintile, comorbidities, visits to primary care in the 6 to 30 months before diagnosis, continuity of care, cancer type and cancer stage at diagnosis, immigrant status was associated with a lower hazard of dying 5-years post-diagnosis for both females (0.7; 95% CI 0.6–0.8) and males (0.7; 95% CI 0.6–0.7) in comparison to long-term residents. This trend held in adjusted models stratified by cancer stage at diagnosis. For example, female immigrants diagnosed with early stage lung cancer had a hazard ratio of 0.5 (95% CI 0.4–0.7) in comparison to long-term residents. Conclusion Overall survival post diagnosis with lung cancer was better among Ontario immigrants versus long-term residents. Additional research, potentially on the protective effects of immigrant enclave and the intersection of immigrant status with racial/ethnic identity, is needed to further explore why better overall survival for immigrants remained.

Abstract In this study, we analyzed pooled data from two prospective population-based cohorts—the Health Retirement Study (HRS) and the English Longitudinal Study of Ageing (ELSA)—to explore the association between trajectories of depressive symptoms and the risk of cardiac events. Depressive symptoms were assessed using the 8-item CES-D scale and categorized into somatic and cognitive-affective subtypes. Trajectories were tracked for four surveys from baseline. Heart disease was identified based on self-reported physician-diagnosed conditions. Hazard ratios and 95% confidence intervals were calculated with Cox proportional risk models that adjusted for potential confounders. In total, 17,787 subjects (59.7% female, median age 63 years) were enrolled at baseline. During a 10-year follow-up, 2409 cases of heart disease were identified. Participants with fluctuating (HR = 1.13, 95% CI: 1.06–1.20), increasing (HR = 1.43, 95% CI: 1.25–1.64), and consistently high (HR = 1.64, 95% CI: 1.45–1.84) depressive symptom trajectories exhibited an increased risk of heart disease compared to those with consistently low depressive symptoms, while a decreasing (HR = 1.07, 95% CI: 0.96–1.19) depressive symptom trajectory did not significantly affect the risk of heart disease. Moreover, the association between heart disease and somatic depressive symptoms was found to be stronger than with cognitive-affective symptoms. These findings suggest a significant link between depressive symptom trajectories and heart disease, with particular emphasis on stronger associations with somatic symptoms. It is recommended that the identification and management of depressive symptoms be incorporated into heart disease prevention strategies.

ABSTRACT: Over 1 year, two KPC-producing and two non–KPC-producing Klebsiella pneumoniae strains were isolated from a patient. Genome and DNA hybridization analyses revealed the first three strains as a clonal lineage, with carbapenem resistance changes due to a Tn2-like transposon on an IncR/IncFII plasmid. The fourth strain, carrying three plasmids, caused a lethal infection and represented a different lineage. All strains belonged to the ST11-SL47-OL101 type. This study highlights the Tn2-like transposon's role in carbapenemase gene spread and the importance of distinguishing between bacterial colonization and infection.

Abstract This study aims to analyze the risk factors associated with delayed postoperative bleeding (DPPB) following colorectal polyp surgery, develop a dynamic nomogram and evaluate the model efficacy, provide a reference for clinicians to identify the patients at high risk of DPPB. Retrospective study was done on patients who underwent endoscopic colorectal polypectomy at the First Hospital of Lanzhou University from January 2020 to March 2023. Differences between the group with and without DPPB were compared, and independent risk factors for DPPB occurrence were identified through univariate analysis and combination LASSO and logistic regression. A dynamic nomogram was constructed based on multiple logistic regression to predict DPPB following colorectal polyp surgery. Model evaluation included receiver operating characteristic (ROC), Calibration curve, Decision curve analysis (DCA). DPPB occurred in 38 of the 1544 patients included. multivariate analysis showed that direct oral anticoagulants (DOACs), polyp location in the right hemi colon, polyp diameter, drink, and prophylactic hemoclips were the independent risk factors for DPPB and dynamic nomogram were established. Model validation indicated area under the ROC curve values of 0.936, 0.796, and 0.865 for the training set, validation set, and full set, respectively. The calibration curve demonstrated a strong alignment between the predictions of the column-line diagram model and actual observations. The decision curve analysis (DCA) displayed a significant net clinical benefit across the threshold probability range of 0–100%. The dynamic nomogram aids clinicians in identifying high-risk patients, enabling personalized diagnosis and treatment.

Abstract The mechanisms underlying the selective regional vulnerability to neurodegeneration in Huntington’s disease (HD) have not been fully defined. To explore the role of astrocytes in this phenomenon, we used single-nucleus and bulk RNAseq, lipidomics, HTT gene CAG repeat-length measurements, and multiplexed immunofluorescence on HD and control post-mortem brains. We identified genes that correlated with CAG repeat length, which were enriched in astrocyte genes, and lipidomic signatures that implicated poly-unsaturated fatty acids in sensitizing neurons to cell death. Because astrocytes play essential roles in lipid metabolism, we explored the heterogeneity of astrocytic states in both protoplasmic and fibrous-like (CD44+) astrocytes. Significantly, one protoplasmic astrocyte state showed high levels of metallothioneins and was correlated with the selective vulnerability of distinct striatal neuronal populations. When modeled in vitro, this state improved the viability of HD-patient-derived spiny projection neurons. Our findings uncover key roles of astrocytic states in protecting against neurodegeneration in HD.

Abstract Background The purpose of this study was to improve the deep learning (DL) model performance in predicting and classifying IMRT gamma passing rate (GPR) by using input features related to machine parameters and a class balancing technique. Methods A total of 2348 fields from 204 IMRT plans for patients with nasopharyngeal carcinoma were retrospectively collected to form a dataset. Input feature maps, including fluence, leaf gap, leaf speed of both banks, and corresponding errors, were constructed from the dynamic log files. The SHAP framework was employed to compute the impact of each feature on the model output for recursive feature elimination. A series of UNet++ based models were trained on the obtained eight feature sets with three fine-tuning methods including the standard mean squared error (MSE) loss, a re-sampling technique, and a proposed weighted MSE loss (WMSE). Differences in mean absolute error, area under the receiver operating characteristic curve (AUC), sensitivity, and specificity were compared between the different models. Results The models trained with feature sets including leaf speed and leaf gap features predicted GPR for failed fields more accurately than the other models (F(7, 147) = 5.378, p

Abstract To retrospectively assess the effectiveness of deep learning (DL) model, based on breast magnetic resonance imaging (MRI), in predicting preoperative lymphovascular invasion (LVI) status in patients diagnosed with invasive breast cancer who have negative axillary lymph nodes (LNs). Data was gathered from 280 patients, including 148 with LVI-positive and 141 with LVI-negative lesions. These patients had undergone preoperative breast MRI and were histopathologically confirmed to have invasive breast cancer without axillary LN metastasis. The cohort was randomly split into training and validation groups in a 7:3 ratio. Radiomics features for each lesion were extracted from the first post-contrast dynamic contrast-enhanced (DCE)-MRI. The Least Absolute Shrinkage and Selection Operator (LASSO) regression method and logistic regression analyses were employed to identify significant radiomic features and clinicoradiological variables. These models were established using four machine learning (ML) algorithms and one DL algorithm. The predictive performance of the models (radiomics, clinicoradiological, and combination) was assessed through discrimination and compared using the DeLong test. Four clinicoradiological parameters and 10 radiomic features were selected by LASSO for model development. The Multilayer Perceptron (MLP) model, constructed using both radiomic and clinicoradiological features, demonstrated excellent performance in predicting LVI, achieving a high area under the curve (AUC) of 0.835 for validation. The DL model (MLP-radiomic) achieved the highest accuracy (AUC = 0.896), followed by DL model (MLP-combination) with an AUC of 0.835. Both DL models were significantly superior to the ML model (RF-clinical) with an AUC of 0.720. The DL model (MLP), which integrates radiomic features with clinicoradiological information, effectively aids in the preoperative determination of LVI status in patients with invasive breast cancer and negative axillary LNs. This is beneficial for making informed clinical decisions.

Abstract Background Recent studies on the association between Helicobacter pylori (H. pylori) infection and obesity have reported conflicting results. Therefore, the purpose of our study was to investigate the association of obesity, abdominal obesity, and metabolic obesity phenotypes with H. pylori infection. Methods A cross-sectional study of 1568 participants aged 20 to 85 was conducted using the National Health and Nutrition Examination Survey (NHANES) cycle 1999–2000. Logistic regression models were employed to evaluate the association of general obesity as defined by body mass index (BMI), abdominal obesity as defined by waist circumference (WC) and waist-height ratio (WHtR), and metabolic obesity phenotypes with H. pylori seropositivity. Subgroup analyses stratified by age were conducted to explore age-specific differences in this association. Results After grouping individuals according to their WHtR, the prevalence rate of WHtR ≥ 0.5 in H. pylori-seropositive participants was significantly higher than that in H. pylori-seronegative participants (79.75 vs. 68.39, P

Abstract The nonconventional yeast Kluyveromyces marxianus has potential for industrial production, but the lack of advanced synthetic biology tools for precise engineering hinders its rapid development. Here, we introduce a CRISPR-Cas9-mediated multilocus integration method for assembling multiple exogenous genes. Using SlugCas9-HF, a high-fidelity Cas9 nuclease, we enhance gene editing precision. Specific genomic loci predisposed to efficient integration and expression of heterologous genes are identified and combined with a set of paired CRISPR-Cas9 expression plasmids and donor plasmids to establish a CRISPR-based biosynthesis toolkit. This toolkit enables genome integration of large gene modules over 12 kb and achieves simultaneous quadruple-locus integration in a single step with 20% efficiency. As a proof-of-concept, we apply the toolkit to screen for gene combinations that promote heme production, revealing the importance of HEM4Km and HEM12Sc. This CRISPR-based toolkit simplifies the reconstruction of complex pathways in K. marxianus, broadening its application in synthetic biology.

Background and aim: Glucocorticoids are the only first-line drugs for severe alcoholic hepatitis (AH), with limited efficacy and various side effects on extrahepatic tissues. Liver-targeting glucocorticoid therapy may have multiple advantages over systemic glucocorticoid for AH. The aim of this study was to determine the role of hepatocellular glucocorticoid receptor (GR) in alcohol-associated steatosis (AS) and AH. Materials and methods: AS was induced by a high-fat diet plus binge alcohol in adult male and female mice with liver-specific knockout (LKO) and heterozygote of GR. AH was induced by chronic-plus-binge in middle-aged male mice with liver-specific knockin of GR. Changes in hepatic mRNA and protein expression were determined by quantitative real-time polymerase chain reaction and Western blot. Results: GR-LKO aggravated steatosis and decreased hepatic expression and circulating levels of albumin in both genders of AS mice but only increased markers of liver injury in male AS mice. Marked steatosis in GR-LKO mice was associated with induction of lipogenic genes and down-regulation of bile acid synthetic genes. Hepatic protein levels of GR, hepatocyte nuclear factor 4 alpha, and phosphorylated signal transducer and activator of transcription 3 were gene-dosage-dependently decreased, whereas that of lipogenic ATP citrate lyase was increased in male GR heterozygote and LKO mice. Interestingly, hepatic expression of estrogen receptor alpha (ERα) was induced, and the essential estrogen-inactivating enzyme sulfotransferase 1e1 was gene-dosage-dependently down-regulated in GR heterozygote and knockout AS mice, which was associated with induction of ERα-target genes. Liver-specific knockin of GR protected against liver injury and steatohepatitis in middle-aged AH mice. Conclusions: Hepatic GR deficiency plays a crucial role in the pathogenesis of AS induced by high-fat diet plus binge, and liver-specific overexpression/activation of GR protects against chronic-plus-binge-induced AH in middle-aged mice. Hepatocellular GR is important for protection against AS and AH.

Abstract Avian influenza A virus H7N9 causes severe human infections with >30% fatality. Currently, there is no H7N9-specific prevention or treatment for humans. Here, from a 2013 H7N9 convalescent case in Hong Kong, we isolate four hemagglutinin (HA)-reactive monoclonal antibodies (mAbs), with three directed to the globular head domain (HA1) and one to the stalk domain (HA2). Two clonally related HA1-directed mAbs, H7.HK1 and H7.HK2, potently neutralize H7N9 and protect female mice from lethal H7N9/AH1 challenge. Cryo-EM structures reveal that H7.HK1 and H7.HK2 bind to a β14-centered surface and disrupt the 220-loop that makes hydrophobic contacts with sialic acid on an adjacent protomer, thereby blocking viral entry. Sequence analysis indicates the lateral patch targeted by H7.HK1 and H7.HK2 to be conserved among influenza subtypes. Both H7.HK1 and H7.HK2 retain HA1 binding and neutralization capacity to later H7N9 isolates from 2016–2017, consistent with structural data showing that the antigenic mutations during this timeframe occur at their epitope peripheries. The HA2-directed mAb H7.HK4 lacks neutralizing activity but when used in combination with H7.HK2 moderately augments female mouse protection. Overall, our data reveal antibodies to a conserved lateral HA1 supersite that confer neutralization, and when combined with a HA2-directed non-neutralizing mAb, augment protection.

Abstract In recombinant protein-producing yeast strains, cells experience high production-related stresses similar to high temperatures. It is possible to increase recombinant protein production by enhancing thermotolerance, but few studies have focused on this topic. Here we aim to identify cellular regulators that can simultaneously activate thermotolerance and high yield of recombinant protein. Through screening at 46 °C, a heat-resistant Kluyveromyces marxianus (K. marxianus) strain FDHY23 is isolated. It also exhibits enhanced recombinant protein productivity at both 30 °C and high temperatures. The CYR1N1546K mutation is identified as responsible for FDHY23’s improved phenotype, characterized by weakened adenylate cyclase activity and reduced cAMP production. Introducing this mutation into the wild-type strain greatly enhances both thermotolerance and recombinant protein yields. RNA-seq analysis reveals that under high temperature and recombinant protein production conditions, CYR1 mutation-induced reduction in cAMP levels can stimulate cells to improve its energy supply system and optimize material synthesis, meanwhile enhance stress resistance, based on the altered cAMP signaling cascades. Our study provides CYR1 mutation as a novel target to overcome the bottleneck in achieving high production of recombinant proteins under high temperature conditions, and also offers a convenient approach for high-throughput screening of recombinant proteins with high yields.

Abstract Developing skeletal editing tools is not a trivial task, and realizing the corresponding single-atom transmutation in a ring system without altering the ring size is even more challenging. Here, we introduce a skeletal editing strategy that enables polycyclic arenols, a highly prevalent motif in bioactive molecules, to be readily converted into N-heteroarenes through carbon–nitrogen transmutation. The reaction features selective nitrogen insertion into the C–C bond of the arenol frameworks by azidative dearomatization and aryl migration, followed by ring-opening, and ring-closing (ANRORC) to achieve carbon-to-nitrogen transmutation in the aromatic framework of the arenol. Using widely available arenols as N-heteroarene precursors, this alternative approach allows the streamlined assembly of complex polycyclic heteroaromatics with broad functional group tolerance. Finally, pertinent transformations of the products, including synthesis complex biheteroarene skeletons, were conducted and exhibited significant potential in materials chemistry.

Anthocyanin (ACN) is a natural antioxidant with multiple biological activities, and the aim of this study was to evaluate the protective effect of ACN on the development and progression of lung cancer and to further explore its possible mechanism of action. In vivo, we fed C57BL/6J mice a 0.5%ACN diet or a control diet to observe their effects on the development and progression of urethane-induced lung cancer. In vitro, multiple lung cancer cell lines were used to investigate the effects of C3G on cell viability. The results showed a reduction in lung tumor burden and downregulation of oxidative phosphorylation and fatty acid degradation pathways in lung tissue of urethane-administrated ACN-fed mice compared with control diet-fed mice. In vitro, cyanidin-3-O-glucoside chloride (C3G) intervention treatment significantly inhibited proliferation and apoptosis of A549 cells. This process is likely due to the modulation of AMPK/mTOR signaling pathway by C3G to regulate cellular fatty acid metabolism and reduce intracellular lipid accumulation which affects the growth of lung cancer cells. These results suggest that ACN can inhibit the development and progression of urethane-induced lung tumors and alter the lipid metabolism of tumors in C57BL/6J mice.

Objective Population-based prevalence estimates of co-occurring mental illness and substance use disorder (herein dual diagnosis) are scarce and derived from a single source (e.g., survey data) which may lead to underestimation of prevalence. We linked administrative data to a representative mental health survey in Ontario, Canada to estimate dual diagnosis prevalence using a data triangulation method. Approach We retrospectively linked the 2002 and 2012 Canadian Community Health Survey on Mental Health (CCHS-MH) to emergency department, inpatient hospital, and outpatient physician records in Ontario, Canada. Mental illness and substance use disorder were ascertained though self-report, Composite International Diagnostic Interview screening, and linked administrative health records. We estimated 1-year, 5-year, and lifetime prevalence of dual diagnosis. Results Of the CCHS-MH survey participants, 14,790 (99.8%) were included in the study. The 1-year, 5-year, and lifetime prevalence of dual diagnosis was 2.3%, 4.8%, and 9.8%, respectively, which attenuated to 2.1%, 4.1%, and 8.4%, respectively, when tobacco use disorder was removed from substance use disorder ascertainment. Conclusion Dual diagnosis is more common in the general population than previously estimated. Considering barriers to accessing care for both mental illness and substance use disorders among people with dual diagnosis annually, our 5-year prevalence estimate is likely most informative for health system planning. Implications In the context of increased health burden, barriers to care access, and challenges to effective treatment associated with dual diagnosis, our findings present a case for increased investment in integrated models of mental healthcare and addiction medicine.

Objectives To examine trends in pregnancy-associated non-fatal and fatal opioid toxicity and all-cause mortality, and identify associated factors. Approach We conducted a population-based study of 1,555,370 pregnancies in Ontario, Canada, 2013-2022. We analyzed linked administrative datasets, including coroner data, and calculated pregnancy-associated (during pregnancy or within one-year post-pregnancy) non-fatal/fatal opioid toxicity and all-cause mortality ratios per 100,000 livebirths by year and timing (pregnancy, post-pregnancy). Poisson regression models analyzed trends in outcomes and generated adjusted relative risks (aRR) of opioid toxicity by socio-demographic and clinical factors. Results Pregnancy-associated non-fatal opioid toxicity increased 220% between 2013 and 2020 (45.5-145.4/100,000 livebirths) before declining by 30% in 2021. Over the study period, fatal opioid toxicity increased 150% (6.8-17.5/100,000) and all-cause mortality increased 120% (32.8-71.2/100,000). Our methods did not identify any opioid toxicity deaths in pregnancy, and most non-fatal (66.6%) and fatal (88.9%) opioid toxicity and all-cause mortality (73.9%) occurred 43-365 days post-pregnancy. The percent of deaths attributed to opioids increased from 12.7% in 2015 to 25.0% in 2020. Substance use disorder (aRR 19.52, 95% CI 16.87-22.58), pre-pregnancy opioid toxicity (aRR 4.69, 3.81-5.78), mental illness (aRR 2.01, 1.75-2.29), high neighbourhood deprivation (aRR 1.45, 1.28-1.64), and social disadvantage (aRR 3.21, 2.77-3.71) were associated with elevated risk of opioid toxicity. Conclusions Pregnancy-associated opioid toxicity and mortality have increased substantially. In 2020, 1 in 4 pregnancy-associated deaths involved opioids. Implications Findings highlight the need for comprehensive care and perinatal harm reduction services. System-level improvements to reduce poor outcomes must include complete data capture of all pregnancy-associated deaths.

Objectives Our understanding of the contribution of new presentations versus pre-existing eating disorders during the COVID-19 pandemic is limited. This study aims to evaluate rates of emergency department (ED) visits and hospitalizations for eating disorders among adolescents and young adults (YA) new to care and those with pre-existing eating disorders during the pandemic. Approach We conducted a population-based cross-sectional study using linked health administrative data for Ontario residents aged 10-26 during the pre-pandemic (Jan. 1, 2017-Feb. 29, 2020) and pandemic periods (Mar. 1, 2020-Jun. 30, 2022). We used Poisson generalized estimating equations models to predict expected overall and monthly rates of eating disorder-related ED visits and hospitalizations among those with a new and pre-existing eating disorder. Results Compared with expected rates, ED visits increased during the pandemic among only adolescents with new eating disorders (adolescent RR 2.12, 95% CI [1.84,2.45]). Additionally, both adolescents and YA with pre-existing eating disorders experienced an increase in ED visits (RR 2.78, 95% CI [2.28, 3.38] and RR 1.52, 95% CI [1.25, 1.85], respectively). Similarly, hospitalizations for new presentations increased solely for adolescents (RR 1.48, 95% CI [1.34,1.64]), while hospitalizations for pre-existing eating disorders increased for both adolescents (RR 1.82, 95% CI [1.43,2.32]) and YA (RR 1.12, 95% CI [1.01,1.23]). Conclusions There was an increase in acute care visits for eating disorders during the pandemic, especially among adolescents and YA with pre-existing conditions. This differentiation is important in advancing our understanding of the pandemic's effects on adolescents and YA and the healthcare system receiving them.

BackgroundEvidence from observational studies on the association between folate and metabolic dysfunction-associated steatotic liver disease (MASLD) is conflicting.AimsThis study aimed to investigate the association between serum folate concentration and MASLD and further assess the causal relationship using Mendelian randomization (MR) analysis.MethodsTo investigate the causal relationship between serum folate and MASLD, we conducted a cross-sectional study that selected 1,117 participants from the 2017–2020 National Health and Nutrition Examination Survey (NHANES). The association between serum folate level and the risk of MASLD was evaluated under a multivariate logistic regression model. In addition, we conducted a two-sample MR study using genetic data from a large genome-wide association study (GWAS) to compare serum folate level (37,465 individuals) and MASLD (primary analysis: 8,434 cases/770,180 controls; Secondary analysis:1,483 cases/17,781 controls) were performed to infer causal relationships between them. Inverse variance weighted (IVW) was used as the primary method of MR Analysis.ResultsThe results from the NHANES database showed that Tertile 3 group (Tertile 3: ≥ 48.6 nmol/L) had a significantly lower risk (OR = 0.58, 95% CI: 0.38–0.88, p = 0.010) of MASLD than Tertile 1 group (Tertile 1: < 22.3 nmol/L) after complete adjustments. However, in the IVW of MR analysis, there was no causal relationship between serum folate level and MASLD risk in the primary analysis (OR = 0.75, 95% CI: 0.55–1.02, p = 0.065) and secondary analysis (OR = 0.83, 95% CI: 0.39–1.74, p = 0.618).ConclusionIn observational analyses, we observed an inverse association between higher serum folate concentrations and a reduced risk of MASLD. Our MR study generated similar results, but the association failed to reach the significance threshold of p

Background: Exposure to ambient fine particulate matter (PM2.5) has been associated with reduced human fecundity. However, the attributable burden has not been estimated for low- and middle-income countries (LMICs), where the exposure–response function between PM2.5 and the infertility rate has been insufficiently studied. Objective: This study examined the associations between long-term exposure to PM2.5 and human fecundity indicators, namely the expected time to pregnancy (TTP) and 12-month infertility rate (IR), and then estimated PM2.5-attributable burden of infertility in LMICs. Methods: We analyzed 164,593 eligible women from 100 Demographic and Health Surveys conducted in 49 LMICs between 1999 and 2021. We assessed PM2.5 exposures during the 12 months before a pregnancy attempt using the global satellite-derived PM2.5 estimates produced by Atmospheric Composition Analysis Group (ACAG). First, we created a series of pseudo-populations with balanced covariates, given different levels of PM2.5 exposure, using a matching approach based on the generalized propensity score. For each pseudo-population, we used 2-stage generalized Gamma models to derive TTP or IR from the probability distribution of the questionnaire-based duration time for the pregnancy attempt before the interview. Second, we used spline regressions to generate nonlinear PM2.5 exposure–response functions for each of the two fecundity indicators. Finally, we applied the exposure–response functions to estimate number of infertile couples attributable to PM2.5 exposure in 118 LMICs. Results: Based on the Gamma models, each 10 µg/m3 increment in PM2.5 exposure was associated with a TTP increase by 1.7 % (95 % confidence interval [CI]: -2.3 %–6.0 %) and an IR increase by 2.3 % (95 %CI: 0.6 %–3.9 %). The nonlinear exposure–response function suggested a robust effect of an increased IR for high-concentration PM2.5 exposure (>75 µg/m3). Based on the PM2.5-IR function, across the 118 LMICs, the number of infertile couples attributable to PM2.5 exposure exceeding 35 µg/m3 (the first-stage interim target recommended by the World Health Organization global air quality guidelines) was 0.66 million (95 %CI: 0.061–1.43), accounting for 2.25 % (95 %CI: 0.20 %–4.84 %) of all couples affected by infertility. Among the 0.66 million, 66.5 % were within the top 10 % high-exposure infertile couples, mainly from South Asia, East Asia, and West Africa. Conclusion: PM2.5 contributes significantly to human infertility in places with high levels of air pollution. PM2.5-pollution control is imperative to protect human fecundity in LMICs.

Abstract Kluyveromyces marxianus is a food‐safe yeast with great potential for producing heterologous proteins. Improving the yield in K. marxianus remains a challenge and incorporating large‐scale functional modules poses a technical obstacle in engineering. To address these issues, linear and circular yeast artificial chromosomes of K. marxianus (KmYACs) were constructed and loaded with disulfide bond formation modules from Pichia pastoris or K. marxianus. These modules contained up to seven genes with a maximum size of 15 kb. KmYACs carried telomeres either from K. marxianus or Tetrahymena. KmYACs were transferred successfully into K. marxianus and stably propagated without affecting the normal growth of the host, regardless of the type of telomeres and configurations of KmYACs. KmYACs increased the overall expression levels of disulfide bond formation genes and significantly enhanced the yield of various heterologous proteins. In high‐density fermentation, the use of KmYACs resulted in a glucoamylase yield of 16.8 g/l, the highest reported level to date in K. marxianus. Transcriptomic and metabolomic analysis of cells containing KmYACs suggested increased flavin adenine dinucleotide biosynthesis, enhanced flux entering the tricarboxylic acid cycle, and a preferred demand for lysine and arginine as features of cells overexpressing heterologous proteins. Consistently, supplementing lysine or arginine further improved the yield. Therefore, KmYAC provides a powerful platform for manipulating large modules with enormous potential for industrial applications and fundamental research. Transferring the disulfide bond formation module via YACs proves to be an efficient strategy for improving the yield of heterologous proteins, and this strategy may be applied to optimize other microbial cell factories.

Abstract Background Posterior cortical atrophy (PCA) is a form of dementia that frequently displays significant visual dysfunction and relatively preserved cognitive and executive functions, thus hindering early diagnosis and treatment. This study aimed to investigate possible fundus markers in PCA patients and compare them with those of typical Alzheimer’s disease (AD) patients to seek potential diagnostic patterns. Methods Age-matched PCA and AD patients and healthy controls (HC) completed optometry, intraocular pressure measurement, neuropsychologic assessments, optical coherence tomography (OCT), and optical coherence tomography angiography (OCTA) examination in one visit. Overall, six outcomes of thicknesses of various retinal layers and seven outcomes of the retinal microvascular network were calculated. After adjusting for age, sex, and years of education, the OCT and OCTA results were analyzed using analysis of covariance and generalized linear models. Correlation analyses were performed using Spearman correlation, and ROC curves were plotted. Results Twelve PCA patients, nineteen AD patients, and thirty HC, aged 45–80 years were included. Fifty HC, thirty AD, and twenty PCA eyes were available for foveal avascular zone (FAZ) area analysis; forty-nine HC, thirty-four AD, and eighteen PCA eyes were available for OCT and OCTA assessments. PCA patients had thinner retinal nerve fiber layer and ganglion cell layer + inner plexiform layer than HC in the 0–3 mm circle and 1–3 mm ring. Few structural differences were observed between the AD group and the other two groups. The flow area of the superficial capillary plexus and the intermediate capillary plexus was smaller in the PCA group than in the HC group in the 0–1 mm circle, 0–3 mm circle. MMSE performed better than any combination of optical parameters in identifying AD and PCA from HC (AUC = 1), while the combination of MoCA, retinal thickness and vascular density of ICP in the 1-3 mm ring, with flow area of ICP in the 0-1 mm circle showed the strongest ability to distinguish PCA from AD (AUC = 0.944). Conclusions PCA patients exhibited similar impairment patterns to AD patients in the fundus structure and microvascular network. OCTA may aid in the non-invasive detection of AD and PCA, but still remains to be substantiated.

Long non-coding RNAs (lncRNAs) have attracted significant scientific attention due to their central role in regulating gene expression and their profound impact on the intricate mechanisms of ovarian function. These versatile molecules exert their influence through various mechanisms, including the coordination of transcription processes, modulation of post-transcriptional events, and the shaping of epigenetic landscapes. Furthermore, lncRNAs function as competitive endogenous RNAs (ceRNAs), engaging in intricate interactions with microRNAs (miRNAs) to finely adjust the expression of target genes. The intricate lncRNA-miRNA-mRNA network serves as a crucial determinant in governing the multifaceted physiological functions of the ovaries. It holds substantial potential in unraveling the causes and progression of reproductive disorders and, importantly, in identifying new therapeutic targets and diagnostic markers for these conditions. A comprehensive comprehension of lncRNAs and their ceRNA activities within the domain of ovarian biology could potentially lead to groundbreaking advancements in clinical interventions and management strategies. This exploration of lncRNAs and their intricate involvement in the regulatory framework provides an extensive platform for deciphering the complex nature of ovarian physiology and pathology. The ongoing progress in this field, which encompasses in-depth investigations into the functional roles of specific lncRNAs, the elucidation of their complex interactions with miRNAs, and the comprehensive profiling of their expression patterns, holds the promise of making significant contributions to our understanding of ovarian biology and reproductive disorders. Ultimately, these breakthroughs will have wide-ranging translational implications, paving the way for the development of precision therapies and personalized medicine strategies to address the myriad challenges in the realm of reproductive health.

Transportation systems design might be supported by participatory approaches refining service design, user experiences, and safety research. The evaluation of mobility alternatives as design improvement strategies requires more advanced instructional techniques with new structures of complexity that can demonstrate, analyze, or compare system performances in order to achieve environmental sustainability in general and the well-being of the end-users in particular. As an emerging technology, games are capable of representing operations and management of passenger freight transportation and increase situational awareness of capacity bottlenecks. However, few research efforts have been devoted to the synthesis of the research progress. Based on the profiling methodology that has been previously employed in retrospect of healthcare simulations, this article presents a bibliometric review of games for health in transportation systems. The study carries out a literature analysis of the modelling methods to assemble gamified scenarios from a planning and managerial perspective. The Web of Science Core Collections are used to identify research items and investigate all the game-based studies in the cross-disciplinary research area of transportation and public health. The technology, game mechanism, and the health-related issue addressed are grouped into broad categories. The research findings are three-fold. (1) The majority of publications are classified into research efforts revolving around slow and road transportation modes. (2) Although scores, badges, and challenges are considered as the frequently seen gamification features, environmental simulation, avatars, and narratives appear as game mechanisms for the activation of the individual decision-making process. (3) Introducing system dynamics, human-computer interaction design, and a stakeholder accountability framework into the multi-stakeholder simulation setting leverages gaming outcomes with health benefits amid transportation system end-users. The literature study concludes with an integrated framework summarizing the best design practices between 2002 and 2022 on how the lessons learned from serious gameplay in virtual environments could translate into real mobility improvements. A selection of future research questions is curated to envisage new links between transportation and health research communities.

Abstract Background The 5´ untranslated region (5´ UTR) plays a key role in regulating translation efficiency and mRNA stability, making it a favored target in genetic engineering and synthetic biology. A common feature found in the 5´ UTR is the poly-adenine (poly(A)) tract. However, the effect of 5´ UTR poly(A) on protein production remains controversial. Machine-learning models are powerful tools for explaining the complex contributions of features, but models incorporating features of 5´ UTR poly(A) are currently lacking. Thus, our goal is to construct such a model, using natural 5´ UTRs from Kluyveromyces marxianus, a promising cell factory for producing heterologous proteins. Results We constructed a mini-library consisting of 207 5´ UTRs harboring poly(A) and 34 5´ UTRs without poly(A) from K. marxianus. The effects of each 5´ UTR on the production of a GFP reporter were evaluated individually in vivo, and the resulting protein abundance spanned an approximately 450-fold range throughout. The data were used to train a multi-layer perceptron neural network (MLP-NN) model that incorporated the length and position of poly(A) as features. The model exhibited good performance in predicting protein abundance (average R2 = 0.7290). The model suggests that the length of poly(A) is negatively correlated with protein production, whereas poly(A) located between 10 and 30 nt upstream of the start codon (AUG) exhibits a weak positive effect on protein abundance. Using the model as guidance, the deletion or reduction of poly(A) upstream of 30 nt preceding AUG tended to improve the production of GFP and a feruloyl esterase. Deletions of poly(A) showed inconsistent effects on mRNA levels, suggesting that poly(A) represses protein production either with or without reducing mRNA levels. Conclusion The effects of poly(A) on protein production depend on its length and position. Integrating poly(A) features into machine-learning models improves simulation accuracy. Deleting or reducing poly(A) upstream of 30 nt preceding AUG tends to enhance protein production. This optimization strategy can be applied to enhance the yield of K. marxianus and other microbial cell factories.

Abstract Background In Switzerland, foreigners account for 25.3% of the permanent resident population, and the fertility rate of migrant women is higher than that of Swiss women. However, migrant women from non-European countries are more likely to report having negative childbirth experiences than Swiss women. For example, during pregnancy, Chinese migrant mothers often felt dissatisfied with the follow-up pregnancy complications and lacked information on medical procedures and prenatal courses. In this paper, we explored their childbirth experiences in Swiss hospitals and how Swiss healthcare providers supported them. Method A qualitative study employing in-depth, semi-structured interviews was conducted with 14 Chinese mothers and 13 family members. All interviews were audio-recorded, transcribed, and translated into English for data analysis. Thematic analysis was employed to generate a detailed description. Results Three main themes were extracted from the transcripts: (1) Sense of security, (2) Intrapartum care, and (3) Postpartum needs. Conclusions Our study shows Chinese migrant mothers prioritized giving birth in a physically and psychologically safe environment, with pain control and practical and emotional support from their intimate partners. They desired a physiological labor and birth with minimal obstetric interventions. Our research also reveals their postpartum needs, emphasizing the importance of postpartum support and obtaining culturally sensitive care during their postpartum hospital stay. The study adds new knowledge of specific migrant studies in Switzerland, as called for by the Swiss Federal Office of Public Health. The results call for the transcultural care skills training of Swiss healthcare providers to enable migrant women to have a more positive childbirth experience.

Summary: Background: Immigrants in high-income countries experienced inequities in COVID-19 severe outcomes. We examined hospitalization and death throughout the pandemic, and change during the vaccine era, in Ontario, Canada. Methods: We conducted a population-based study using linked immigration and health data, following two cohorts for 20 months from January 1, 2020 (pre-vaccine) and September 1, 2021 (vaccine era). We used multivariable Poisson generalized estimating equation regression to estimate adjusted rate ratios (aRR) with 95% confidence intervals (CI), accounting for age, sex and co-morbidities. We calculated age-standardized years of life lost (ASYRs) rates by immigrant category. Findings: Of 11,692,387 community-dwelling adults in the pre-vaccine era cohort and 11,878,304 community-dwelling adults in the vaccine era cohort, 21.6% and 21.4% of adults in each era respectively were immigrants. Females accounted for 57.9% and 57.8% of sponsored family, and 68.4% and 67.6% of economic caregivers, in each era respectively. Compared to other Ontarians in the pre-vaccine era cohort, hospitalization rates were highest for refugees (aRR [95% CI] 3.41 [3.39–3.44]) and caregivers (3.13 [3.07–3.18]), followed by sponsored family and other economic immigrants. Compared to other Ontarians, aRRs were highest for immigrants from Central America (5.00 [4.92–5.09]), parts of South Asia (3.95 [3.89–4.01]) and Jamaica (3.56 [3.51–3.61]) with East Asians having lower aRRs. Mortality aRRs were similar to hospitalization aRRs. In the vaccine era, all aRRs were attenuated and most were similar to or lower than other Ontarians, with refugees and a few regions maintaining higher rates. In the pre-vaccine era ASYRs were higher for all immigrant groups. ASYRs dropped in the vaccine era with only refugees continuing to have higher rates. Interpretation: Immigrants, particularly refugees, experienced greater premature mortality. aRRs for most immigrant groups dropped substantially after high vaccine coverage was achieved. Vaccine outreach and improvements in the social determinants of health are needed. Funding: Canadian Institutes of Health Research, Canada Research Chairs Program.

Exposure to anesthesia in early life may cause severe damage to the brain and lead to cognitive impairment. The underlying mechanisms, which have only been investigated in a limited scale, remains largely elusive. We performed translatome and transcriptome sequencing together for the first time in hippocampus of neonatal mice that were exposed to sevoflurane. We treated a group of neonatal mice with 2.5 % sevoflurane for 2 h on day 6, 7, 8, 9 and treated another group on day 6, 7. We performed behavioral study after day 30 for both groups and the control to evaluate the cognitive impairment. On day 36, we collected translatome and transcriptome from the hippocampus in the two groups, compared the gene expression levels between the groups and the control, and validated the results with RT-qPCR. We identified 1750 differentially expressed genes (DEGs) from translatome comparison and 1109 DEGs from transcriptome comparison. As expected, translatome-based DEGs significantly overlapped with transcriptome-based DEGs, and functional enrichment analysis generated similar enriched cognition-related GO terms and KEGG pathways. However, for many genes like Hspa5, their alterations in translatome differed remarkably from those in transcriptome, and Western blot results were largely concordant with the former, suggesting that translational regulation plays a significant role in cellular response to sevoflurane. Our study revealed global alterations in translatome and transcriptome of mice hippocampus after neonatal exposure to sevoflurane anesthesia and highlighted the importance of translatome analysis in understanding the mechanisms responsible for anesthesia-induced cognitive impairment.

This study conducted a comparative analysis of extractable hopanoid hydrocarbons and those released via stepwise pyrolysis of typical Eocene immature oil shales in China, namely the Huadian, Maoming, and Fushun shales. Both the Huadian and Maoming shales exhibit immature indicators in extractable and kerogen-bound hopanoids (notably, high abundance of C29 to C32 17β,21β-hopanes and unsaturated hopenes). In contrast, the Fushun oil shale's hopanoids from extracts and pyrolyzates suggest a higher maturity level. The absence of neohopenes in the pyrolyzates of the shales underpins that the kerogen-bound hopanoid skeletons resist rearrangement. However, the Huadian oil shale's asymmetric distribution of C29 and C30 hopenes and neohopenes hints at the presence of an additional source. Novel unsaturated hopenes, such as hop-20 (21)-enes, identified in pyrolyzates of the three kerogens at various pyrolysis temperatures, reveal the occurrence of double bonds in kerogen-bound hopanoid skeletons without methyl rearrangements. The absence of hop-20-(21)-ene in extracts suggests that it might act as an intermediate of these novel hopenes during the epimerization of hopanoid skeletons within kerogen. The extractable and pyrolytic hopanoids' stereochemical alignment indicates that epimerization may occur in both ring systems and alkyl side chains of kerogen-bound hopanoid skeletons. Sequential stepwise pyrolysis proves to be a quick screening method for geological hopanoids without causing any significant alteration to the original skeletons even when cracking multiple covalent bonds is necessary.

AIM: To investigate the effect of morroniside (Mor) on lipopolysaccharide (LPS)-treated iris pigment epithelial cells (IPE). METHODS: IPE cells were induced by LPS and treated with Mor. Cell proliferation was detected by cell counting kit (CCK) -8, apoptosis was detected by flow cytometry, the levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-6, and IL-8 were measured by enzyme-linked immunosorbent assay (ELISA) kits, and the protein expression of TLR4, JAK2, p-JAK2, STAT3, and p-STAT3 was analyzed by Western blotting. In addition, overexpression of TLR4 and Mor treatment of LPS-stimulated IPE cells were also tested for the above indices. RESULTS: Mor effectively promoted the proliferation and inhibited the apoptosis of LPS-treated IPE cells. In addition, Mor significantly reduced the levels of TNF-α, IL-6, and IL-8 and significantly inhibited the expression of TLR4, p-JAK2, and p-STAT3 in LPS-treated IPE cells. The effect of Mor on LPS-treated IPE cells was markedly attenuated after overexpression of TLR4. CONCLUSION: These findings suggest that Mor may ameliorate LPS-induced inflammatory damage and apoptosis in IPE through inhibition of TLR4/JAK2/STAT3 pathway.

Abstract Background Early detection of hepatocellular carcinoma (HCC) is important in order to improve patient prognosis and survival rate. Methylation sequencing combined with neural networks to identify cell-free DNA (cfDNA) carrying aberrant methylation offers an appealing and non-invasive approach for HCC detection. However, some limitations exist in traditional methylation detection technologies and models, which may impede their performance in the read-level detection of HCC. Methods We developed a low DNA damage and high-fidelity methylation detection method called No End-repair Enzymatic Methyl-seq (NEEM-seq). We further developed a read-level neural detection model called DeepTrace that can better identify HCC-derived sequencing reads through a pre-trained and fine-tuned neural network. After pre-training on 11 million reads from NEEM-seq, DeepTrace was fine-tuned using 1.2 million HCC-derived reads from tumor tissue DNA after noise reduction, and 2.7 million non-tumor reads from non-tumor cfDNA. We validated the model using data from 130 individuals with cfDNA whole-genome NEEM-seq at around 1.6X depth. Results NEEM-seq overcomes the drawbacks of traditional enzymatic methylation sequencing methods by avoiding the introduction of unmethylation errors in cfDNA. DeepTrace outperformed other models in identifying HCC-derived reads and detecting HCC individuals. Based on the whole-genome NEEM-seq data of cfDNA, our model showed high accuracy of 96.2%, sensitivity of 93.6%, and specificity of 98.5% in the validation cohort consisting of 62 HCC patients, 48 liver disease patients, and 20 healthy individuals. In the early stage of HCC (BCLC 0/A and TNM I), the sensitivity of DeepTrace was 89.6 and 89.5% respectively, outperforming Alpha Fetoprotein (AFP) which showed much lower sensitivity in both BCLC 0/A (50.5%) and TNM I (44.7%). Conclusions By combining high-fidelity methylation data from NEEM-seq with the DeepTrace model, our method has great potential for HCC early detection with high sensitivity and specificity, making it potentially suitable for clinical applications. DeepTrace: https://github.com/Bamrock/DeepTrace

Abstract Background Metastasis is still a major cause of poor pathological outcome and prognosis in esophageal squamous cell carcinoma (ESCC) patients. NUAK1 has been reported highly expressed in many human cancers and is associated with the poor prognosis of cancer patients. However, the role of NUAK1 and its underlying signaling mechanism in ESCC metastasis remain unclear. Methods Expression of NUAK1 in ESCC was detected by real-time quantitative RT-PCR (qRT-PCR), Western blotting and immunohistochemical staining. MTT, colony formation, wound-healing and transwell assays were used to determine the role NUAK1 in vitro. Metastasis was evaluated by use of an experimental pulmonary metastasis model in BALB/c-nu/nu mice. The mechanisms were assessed by using coimmunoprecipitation, immunofluorescence and dual-luciferase reporter gene experiments. Results NUAK1 was highly expressed in ESCC tissues compared with the adjacent normal esophageal epithelial tissues. Moreover, the elevated expression of NUAK1 positively correlated with tumor invasion depth, lymph node metastasis, pathological TNM stage, and poor survival in ESCC patients. Further experiments showed that NUAK1 overexpression did not change the cell viability and colony formation of ESCC cells, while remarkably promoted the migration and invasion in vitro and experimental pulmonary metastasis in vivo. Mechanistically, NUAK1 enhanced the transcription level of Slug, which enhanced the migratory and invasive capability of ESCC cells. Consistently, silencing Slug almost completely diminished the migration and invasion of NUAK1-overexpressing ESCC cells. Further studies demonstrated that NUAK1 upregulated the transcription activity of Slug through activating the JNK/c-Jun pathway. Conclusion These results demonstrated that NUAK1 promoted the metastasis of ESCC cells through activating JNK/c-Jun/Slug signaling, indicating NUAK1 is a promising therapeutic target for metastatic ESCC.

Abstract Objective To explore the grief experiences of family members of patients with advanced malignant tumors before and after death. Methods This study used both quantitative and qualitative research methods. A total of 10 people with family members with terminal malignant tumors were chosen and assessed five times according to a specific non-invasive mortality index and the Distress Thermometer scale. Additionally, the participants attended an in-depth interview. Results The grief experiences of the bereaved included their knowledge of and attitude towards death, the physical and mental conditions of the family members of patients in the terminal stage, the needs of family members, and the response to death and growth of those family members. Conclusions The grief experience interviews of family members of patients with advanced malignant tumors are universal. It is suggested that the nursing staff should pay attention to the emotional experience of the bereaved after the death of the patient throughout the whole nursing process, including the continuous follow-up during the home period. It is hoped that the implementation of grief counseling methods in the later stage can help the bereaved to successfully go through the grieving period, prevent grief disorders, and help them return to society.

The fault diagnosis of rotating machinery is vital in industry but traditionally depends on manual expertise, requiring substantial resources. To improve diagnostic accuracy, enable effective condition monitoring, and minimize the impact of faults on operations, advanced diagnostic techniques are essential. Hence, we propose an advanced fault diagnosis framework that leverages improved particle swarm optimization (IPSO), variational mode decomposition (VMD), and probabilistic neural networks (PNN) to accurately diagnose faults in rotating machinery using gear and rolling bearing vibration signals. Initially, the vibration signals are decomposed into intrinsic mode functions via VMD, enabling the capture of subtle but critical fault features. To address parameter selection challenges in VMD, we employed IPSO to optimize the VMD parameters, ensuring the optimal decomposition effect. Further, we refined the feature set by applying Laplace fraction optimization and feature dimensionality reduction, isolating sensitive features that serve as input to a PNN-based fault classification model. Experimental results demonstrated that this IPSO-VMD-PNN framework achieves high diagnostic accuracy for various fault types, establishing it as an effective tool for fault identification in rotating machinery.

As the transmission component in mechanical systems, hydrodynamic journal bearing is widely used in large electromechanical equipment. The instability of the hydrodynamic journal bearing often results in mechanical wear and damage, which may cause maintenance shutdowns and significant financial losses. Accordingly, assessing the hydrodynamic journal bearing online is a highly effective approach to guaranteeing reliability. To analyze oil film thickness distribution under various assembly characteristics, this paper proposes a Fluid–Solid coupling analysis method based on hydrodynamic lubrication theory and Hertzian elastic deformation theory. The novelty is that the difference between the oil film thicknesses at the same measuring angle within the two opposing faces of the bearing shell is taken as the assessment parameter, which can directly indicate the deviation degree of the stator axis. Comparison of simulation and experimental results validated that the proposed method exhibits exceptional accuracy in practical applications.

Accurately measuring drug and its release kinetics in both in vitro and in vivo environments is crucial for enhancing therapeutic effectiveness while minimizing potential side effects. Nevertheless, the real-time visualization of drug release from microspheres to monitor potential overdoses remains a challenge. The primary objective of this investigation was to employ fluorescence imaging for the real-time monitoring of drug release from microspheres in vitro, thereby simplifying the laborious analysis associated with the detection of drug release. Two distinct varieties of microspheres were fabricated, each encapsulating different drugs within PLGA polymers. Cy5 was selected as the donor, and Cy7 was selected as the acceptor for visualization and quantification of the facilitated microsphere drug release through the application of the fluorescence resonance energy transfer (FRET) principle. The findings from the in vitro experiments indicate a correlation between the FRET fluorescence alterations and the drug release profiles of the microspheres.

BackgroundMaternal pushing during the second stage of labor could influence labor progress and maternal-neonatal outcomes. Although the image of health care providers directing the laboring women to push during the second stage of labor could be commonly observed globally, this practice is not sufficiently researched and is questioned regarding its effectiveness and outcomes on the mother and baby. Meanwhile, a strategy referred to as “spontaneous pushing,” which supports women to push by following their bodily urges, has been evaluated in several trials. However, in China, spontaneous pushing is not common practice. Notwithstanding the evaluation of spontaneous pushing, there is a lack of high-quality evidence to support either strategies of directed pushing or spontaneous pushing. ObjectiveThis study aims to test the feasibility of a future randomized controlled trial to compare the effects of spontaneous pushing and directed pushing during the second stage of labor for maternal and neonatal outcomes in China. MethodsA nonrandomized, single-group, noninferiority feasibility study will be conducted in a public hospital in Hebei Province, China. In total, 105 women meeting the selection criteria will be recruited to receive the intervention (spontaneous pushing), while 105 sets of medical notes from women who received routine care (directed pushing) will be identified and reviewed to compare outcomes for both cohorts. A mixed methods approach will be used to assess primary outcomes (feasibility and acceptability) and secondary outcomes (effectiveness). ResultsData collection took place between May and October 2023. A total of 110 women were invited to participate in the intervention of spontaneous pushing. Midwives’ interviews were conducted and will be transcribed for analysis in March 2024. The data analysis is planned to be completed by May 2024. ConclusionsThis feasibility study will provide important information by conducting a full-scale clinical trial in the future as well as the potential facilitators and barriers of it. A future randomized controlled trial is likely to have considerable policy and funding impacts regarding pushing management during the second stage of labor and improvement in women’s childbirth experience. Trial RegistrationChinese Clinical Trial Register ChiCTR2300071178; https://tinyurl.com/mudtnbft International Registered Report Identifier (IRRID)DERR1-10.2196/55701

Background: Glucocorticoids (GCs) are widely used in the treatment of inflammatory liver diseases and sepsis, but GC’s various side effects on extrahepatic tissues limit their clinical benefits. Liver-targeting GC therapy may have multiple advantages over systemic GC therapy. The purpose of this study was to develop novel liver-targeting GC prodrugs as improved treatment for inflammatory liver diseases and sepsis. Methods: A hydrophilic linker or an ultra-hydrophilic zwitterionic linker carboxylic betaine (CB) was used to bridge cholic acid (CA) and dexamethasone (DEX) to generate transporter-dependent liver-targeting GC prodrugs CA-DEX and the highly hydrophilic CA-CB-DEX. The efficacy of liver-targeting DEX prodrugs and DEX were determined in primary human hepatocytes (PHH), macrophages, human whole blood, and/or mice with sepsis induced by cecal ligation and puncture. Results: CA-DEX was moderately water soluble, whereas CA-CB-DEX was highly water soluble. CA-CB-DEX and CA-DEX displayed highly transporter-dependent activities in reporter assays. Data mining found marked dysregulation of many GR-target genes important for lipid catabolism, cytoprotection, and inflammation in patients with severe alcoholic hepatitis. These key GR-target genes were similarly and rapidly (within 6 h) induced or down-regulated by CA-CB-DEX and DEX in PHH. CA-CB-DEX had much weaker inhibitory effects than DEX on endotoxin-induced cytokines in mouse macrophages and human whole blood. In contrast, CA-CB-DEX exerted more potent anti-inflammatory effects than DEX in livers of septic mice. Conclusions: CA-CB-DEX demonstrated good hepatocyte-selectivity in vitro and better anti-inflammatory effects in vivo. Further test of CA-CB-DEX as a novel liver-targeting GC prodrug for inflammatory liver diseases and sepsis is warranted.

Soy leghemoglobin, when bound to heme, imparts a meat-like color and flavor and can serve as a substitute for animal-derived proteins. Enhancing cellular heme synthesis improves the recombinant expression of leghemoglobin in yeast. To achieve high-level expression of leghemoglobin A (LBA) in Kluyveromyces marxianus, a food-safe yeast, large-scale heme synthesis modules were transferred into K. marxianus using yeast artificial chromosomes (KmYACs). These modules contained up to 8 native and heterologous genes to promote the supply of heme precursors and downstream synthesis. Next, eight genes inhibiting heme or LBA synthesis were individually or combinatorially deleted, with the lsc1Δssn3Δ mutant yielding the best results. Subsequently, heme synthesis modules were combined with the lsc1Δssn3Δ mutant. In the resulting strains, the module genes were all actively expressed. Among these module genes, heterologous S. cerevisiae genes in the downstream heme synthesis pathway significantly enhanced the expression of their counterparts in K. marxianus, resulting in high heme content and LBA yield. After optimizing the medium recipe by adjusting the concentrations of glucose, glycine, and FeSO4·7H2O, a heme content of 66.32 mg/L and an intracellular LBA titer of 7.27 g/L were achieved in the engineered strain in a 5 L fermentor. This represents the highest intracellular expression of leghemoglobin in microorganisms to date. The leghemoglobin produced by K. marxianus can be utilized as a safe ingredient for plant-based protein products.

Background. Data on the economic burden of chronic hepatitis C (CHC) among immigrants are limited. Our objective was to estimate the CHC-attributable mortality and healthcare costs among immigrants in Ontario, Canada. Methods. We conducted a population-based matched cohort study among immigrants diagnosed with CHC between May 31, 2003, and December 31, 2018, using linked health administrative data. Immigrants with CHC (exposed) were matched 1 : 1 to immigrants without CHC (unexposed) using a combination of hard (index date, sex, and age) and propensity-score matching. Net costs (2020 Canadian dollars) collected from the healthcare payer perspective were calculated using a phase-of-care approach and used to estimate long-term costs adjusted for survival. Results. We matched 5,575 exposed individuals with unexposed controls, achieving a balanced match. The mean age was 47 years, and 52% was male. On average, 10.5% of exposed and 3.5% of unexposed individuals died 15 years postindex (relative risk = 2.9; 95% confidence interval (CI): 2.6–3.5). The net 30-day costs per person were $88 (95% CI: 55 to 122) for the prediagnosis, $324 (95% CI: 291 to 356) for the initial phase, $1,016 (95% CI: 900 to 1,132) for the late phase, and $975 (95% CI: −25 to 1,974) for the terminal phase. The mean net healthcare cost adjusted for survival at 15 years was $90,448. Conclusions. Compared to unexposed immigrants, immigrants infected with CHC have higher mortality rates and greater healthcare costs. These findings will support the planning of HCV elimination efforts among key risk groups in the province.

Abstract The human microbiome plays a crucial role in maintaining health, with advances in high‐throughput sequencing technology and reduced sequencing costs triggering a surge in microbiome research. Microbiome studies generally incorporate five key phases: design, sampling, sequencing, analysis, and reporting, with sequencing strategy being a crucial step offering numerous options. Present mainstream sequencing strategies include Amplicon sequencing, Metagenomic Next‐Generation Sequencing (mNGS), and Targeted Next‐Generation Sequencing (tNGS). Two innovative technologies recently emerged, namely MobiMicrobe high‐throughput microbial single‐cell genome sequencing technology and 2bRAD‐M simplified metagenomic sequencing technology, compensate for the limitations of mainstream technologies, each boasting unique core strengths. This paper reviews the basic principles and processes of these three mainstream and two novel microbiological technologies, aiding readers in understanding the benefits and drawbacks of different technologies, thereby guiding the selection of the most suitable method for their research endeavours.

Abstract The development of high-throughput omics technology has greatly promoted the development of biomedicine. However, the poor reproducibility of omics techniques limits their application. It is necessary to use standard reference materials of complex RNAs or proteins to test and calibrate the accuracy and reproducibility of omics workflows. The transcriptome and proteome of most cell lines shift during culturing, which limits their applicability as standard samples. In this study, we demonstrated that the human hepatocellular cell line MHCC97H has a very stable transcriptome (r = 0.983~0.997) and proteome (r = 0.966~0.988 for data-dependent acquisition, r = 0.970~0.994 for data-independent acquisition) after 9 subculturing generations, which allows this steady standard sample to be consistently produced on an industrial scale in long term. Moreover, this stability was maintained across labs and platforms. In sum, our study provides omics standard reference material and reference datasets for transcriptomic and proteomics research. This helps to further standardize the workflow and data quality of omics techniques and thus promotes the application of omics technology in precision medicine.

The drive at the center of gravity (DCG) concept-based twin-ball-screw drive mechanism (TBSDM) is vital in automated factories for its robustness and reliability. However, changes in the worktable mass or position result in changes in the center of gravity (CG), significantly affecting the system’s dynamic properties. In this regard, this paper introduces a novel analytical model using improved receptance coupling to analyze vibrations in four modes. A mathematical framework for the twin TBSDM is generated, and the effect of changing the worktable position–mass on each mode is examined. The applicability of the proposed method is verified based on dynamic experiments that were carried out on a TBSDM of a CNC grinding wheel machine tool. After thoroughly analyzing the experimental and theoretical results, it is revealed that changing the worktable position primarily influences the rotational and axial vibrations of the twin ball screw (TBS). Furthermore, changes in the worktable mass significantly affect the coupling vibration among the TBSs and rotors or bearings. Moreover, in terms of performance, the variances between the theoretical and experimental natural frequencies are consistently below 5%. Thus, the proposed method is promising for the improvement of the modeling and analysis of the TBSDM.

This article explores the intricacies of micro-grinding parameter control for hard and brittle materials, with a specific focus on Zirconia ceramics (ZrO2) and Optical Glass (BK7). Given the increasing demand and application of these materials in various high-precision industries, this study aims to provide a comprehensive kinematic analysis of material removal during the micro-grinding process. According to the grinding parameters selected to be analyzed in this study, the ac-max values are between (9.55 nm ~ 67.58 nm). Theoretical modeling of the grinding force considering the brittle and ductile removal phase, frictional effects, the possibility of grit to cut materials, and grinding conditions is very important in order to control and optimize the surface grinding process. This research introduces novel models for predicting and optimizing micro-grinding forces effectively. The primary objective is to establish a micro-grinding force model that facilitates the easy manipulation of micro-grinding parameters, thereby optimizing the machining process for these challenging materials. Through experimental investigations conducted on Zirconia ceramics, the paper evaluates a mathematical model of the grinding force, highlighting its significance in predicting and controlling the forces involved in micro-grinding. The suggested model underwent thorough testing to assess its validity, revealing an accuracy with average variances of 6.616% for the normal force and 5.752% for the tangential force. Additionally, the study delves into the coefficient of friction within the grinding process, suggesting a novel frictional force model. This model is assessed through a series of experiments on Optical Glass BK7, aiming to accurately characterize the frictional forces at play during grinding. The empirical results obtained from both sets of experiments—on Zirconia ceramics and Optical Glass BK7—substantiate the efficacy of the proposed models. These findings confirm the models’ capability to accurately describe the force dynamics in the micro-grinding of hard and brittle materials. The research not only contributes to the theoretical understanding of micro-grinding processes but also offers practical insights for enhancing the efficiency and effectiveness of machining operations involving hard and brittle materials.

A circularly polarized wearable antenna operating in the 2.45/5.8 GHz ISM dual bands is proposed, which consists of a coplanar waveguide-fed rectangular monopole antenna and two parasitic branches. The monopole rectangular radiation patch can generate 2.45 and 7 GHz frequency bands and has linear polarization characteristics. By adding L-shaped parasitic branches and L-like grounded branches on both sides of the monopole, the 7 GHz operating frequency band can be moved to the 5.8 GHz frequency band, and circular polarization characteristics can be achieved in both the 2.45 and 5.8 GHz frequency bands. The size of the antenna is 48.7 mm × 42.1 mm × 1.016 mm. The simulated −10 dB impedance bandwidths of the antenna are 1.8–2.66 GHz and 5.48–6.98 GHz, respectively. The 3 dB axial ratio bandwidths are 2.34–2.67 GHz and 5.58–6.39 GHz, respectively, and it has dual-band circular polarization characteristics. In addition, the radiation characteristics of the antenna and its safety performance near the human body were analyzed. The antenna prototype has been constructed, and the measurement and simulation results have good consistency. The proposed antenna is suitable for application in wearable devices.

Background Total body water (TBW) increases with growth, but the body water percentage (TBW%) decreases with aging. The objective of our study was to delineate TBW% in males and females by bioelectrical impedance analysis (BIA) from early childhood to old age. Methods We enrolled 545 participants aged 3 to 98 years (258 male, 287 female). Among the participants, 256 had a normal weight and 289 were overweight. The TBW was measured by BIA, and TBW% was derived by dividing the TBW (L) value by body weight (kg). For analysis, we divided participants into the four age groups of 3–10, 11–20, 21–60, and ≥61 years. Results In normal-weight subjects, the TBW% was similar at 62% between males and females in the 3–10-year group. It remained unchanged in males until and through adult life, then decreased to 57% in the ≥61-year group. In normal-weight female subjects, the TBW% decreased to 55% in the 11–20-year group, remained relatively unaltered in the 21–60-year group, then decreased to 50% in the ≥61-year group. In overweight subjects, the TBW% values in males, as well as females, were significantly lower as compared to those with normal weight. Conclusion Our study showed that the TBW% in normal-weight males changes very little from early childhood to adult life compared to that of females, who showed a decrease in TBW% during the pubertal years. In normal-weight subjects of both sexes, the TBW% decreased after the age of 60 years. Overweight subjects had significantly lower TBW% as compared to those with normal weight.

Abstract Background The pathogenic viruses transmitted by mosquitoes cause a variety of animal and human diseases and public health concerns. Virome surveillance is important for the discovery, and control of mosquito-borne pathogenic viruses, as well as early warning systems. Virome composition in mosquitoes is affected by mosquito species, food source, and geographic region. However, the complex associations of virome composition remain largely unknown. Results Here, we profiled the high-depth RNA viromes of 15 species of field-caught adult mosquitoes, especially from Culex, Aedes, Anopheles, and Armigeres in Hainan Island from 2018 to 2020. We detected 57 known and 39 novel viruses belonging to 15 families. We established the associations of the RNA viruses with mosquito species and their foods, indicating the importance of feeding acquisition of RNA viruses in determining virome composition. A large fraction of RNA viruses were persistent in the same mosquito species across the 3 years and different locations, showing the species-specific stability of viromes in Hainan Island. In contrast, the virome compositions of single mosquito species in different geographic regions worldwide are visibly distinct. This is consistent with the differences in food sources of mosquitoes distributed broadly across continents. Conclusions Thus, species-specific viromes in a relatively small area are limited by viral interspecific competition and food sources, whereas the viromes of mosquito species in large geographic regions may be governed by ecological interactions between mosquitoes and local environmental factors. Video Abstract

Abstract Glioblastoma (GBM) diffusely infiltrates the brain and intermingles with non-neoplastic brain cells, including astrocytes, neurons and microglia/myeloid cells. This complex mixture of cell types forms the biological context for therapeutic response and tumor recurrence. We used single-nucleus RNA sequencing and spatial transcriptomics to determine the cellular composition and transcriptional states in primary and recurrent glioma and identified three compositional ‘tissue-states’ defined by cohabitation patterns between specific subpopulations of neoplastic and non-neoplastic brain cells. These tissue-states correlated with radiographic, histopathologic, and prognostic features and were enriched in distinct metabolic pathways. Fatty acid biosynthesis was enriched in the tissue-state defined by the cohabitation of astrocyte-like/mesenchymal glioma cells, reactive astrocytes, and macrophages, and was associated with recurrent GBM and shorter survival. Treating acute slices of GBM with a fatty acid synthesis inhibitor depleted the transcriptional signature of this pernicious tissue-state. These findings point to therapies that target interdependencies in the GBM microenvironment.

Linezolid combined with rifampicin has shown excellent clinical outcomes against infection by multi-resistant Gram-positive bacteria. However, several studies have indicated that rifampicin reduces the plasma concentration of linezolid in patients with severe infection. Linezolid has been recommended for the treatment of patients with multidrug-resistant or extensively drug-resistant tuberculosis. However, studies on the interaction between linezolid and rifampicin in patients suffering from tuberculosis with infection are lacking. We evaluated the interaction between linezolid and rifampicin based on therapeutic drug monitoring (TDM). A retrospective analysis was undertaken for patients with tuberculosis and infection who were treated with linezolid and undergoing TDM. Patients were divided into the linezolid group and linezolid + rifampicin group. Data on demographic characteristics, disease, duration of linezolid therapy, and the plasma concentration of linezolid were used for statistical analyses. Eighty-eight patients with tuberculosis and infection were assessed. Values for the peak (Cmax) and trough (Cmin) concentrations of linezolid in plasma were available for 42 and 46 cases, respectively. Patients in the linezolid group had a significantly higher Cmax [15.76 (8.07–26.06) vs. 13.18 (7.48–23.64) mg/L, p = 0.048] and Cmin [8.38 (3.06–16.53) vs. 4.27 (0.45–10.47), p = 0.005] than those in the linezolid + rifampicin group. The plasma concentration of linezolid increased obviously in two patients after rifampicin discontinuation. However, the total efficiency and prevalence of hematologic adverse reactions were not significantly different in the linezolid group and linezolid + rifampin group. The plasma concentration of linezolid decreased upon combination with rifampicin, suggesting that TDM may aid avoidance of subtherapeutic levels of linezolid upon co-treatment with rifampicin.