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Abstract The detection of lymph node (LN) involvement is fundamental for staging colorectal cancer (CRC) and aids in clinical decision‐making. Traditionally, determining LN status predominantly relies heavily on histological examination of LN specimens, which can occasionally lead to overtreatment. This study aims to develop a clinical prediction model using machine learning algorithms to assess the risk of mesenteric LN metastasis preoperatively, based on computed tomography images and clinicopathological data from CRC patients. Our findings demonstrate that the predictive model based on XGBoost algorithms exhibited the optimal performance, with area under the curve values consistently stable across training (0.836, 95% confidence interval [CI]: 0.750–0.902) and validation (0.831, 95% CI: 0.688–0.927) cohorts. The model was further elucidated using SHapley Additive Explanation values, which ranked predictors in the XGBoost model by their importance, providing insights into the model's decision‐making process. Additionally, the force plot visualizes the contribution of each variable to the prediction for individual samples. The as‐obtained model may have the potential to aid in clinical treatment planning, optimize the selection of surgical methods, and guide the decision‐making process for adjuvant therapy before surgery.

Abstract Pathological data showed focal inflammation and regions of diffuse neuronal loss in the cortex of people with multiple sclerosis (MS). In this work, we applied a novel model (“soma and neurite density imaging (SANDI)”) to multishell diffusion-weighted MRI data acquired in healthy subjects and people with multiple sclerosis (pwMS), in order to investigate inflammation and degeneration-related changes in the cortical tissue of pwMS. We aimed to (i) establish whether SANDI is applicable in vivo clinical data; (ii) investigate inflammatory and degenerative changes using SANDI soma fraction (f soma )—a marker of cellularity—in both cortical lesions and in the normal-appearing-cortex and (iii) correlate SANDI f soma with clinical and biological measures in pwMS. We applied a simplified version of SANDI to a clinical scanners. We then provided evidence that pwMS exhibited an overall decrease in cortical SANDI f soma compared to healthy subjects, suggesting global degenerative processes compatible with neuronal loss. On the other hand, we have found that progressive pwMS showed a higher SANDI f soma in the outer part of the cortex compared to relapsing–remitting pwMS, possibly supporting current pathological knowledge of increased innate inflammatory cells in these regions. A similar finding was obtained in subpial lesions in relapsing–remitting patients, reflecting existing pathological data in these lesion types. A significant correlation was found between SANDI f soma and serum neurofilament light chain—a biomarker of inflammatory axonal damage—suggesting a relationship between SANDI soma fraction and inflammatory processes in pwMS again. Overall, our data show that SANDI f soma is a promising biomarker to monitor changes in cellularity compatible with neurodegeneration and neuroinflammation in the cortex of MS patients.

IntroductionAcute myeloid leukemia (AML) is an aggressive blood cancer with high heterogeneity and poor prognosis. Although the metabolic reprogramming of nicotinamide adenine dinucleotide (NAD) has been reported to play a pivotal role in the pathogenesis of acute myeloid leukemia (AML), the prognostic value of NAD metabolism and its correlation with the immune microenvironment in AML remains unclear.MethodsWe utilized our large-scale RNA-seq data on 655 patients with AML and the NAD metabolism-related genes to establish a prognostic NAD metabolism score based on the sparse regression analysis. The signature was validated across three independent datasets including a total of 1,215 AML patients. ssGSEA and ESTIMATE algorithms were employed to dissect the tumor immune microenvironment. Ex vivo drug screening and in vitro experimental validation were performed to identify potential therapeutic approaches for the high-risk patients. In vitro knockdown and functional experiments were employed to investigate the role of SLC25A51, a mitochondrial NAD+ transporter gene implicated in the signature.ResultsAn 8-gene NAD metabolism signature (NADM8) was generated and demonstrated a robust prognostic value in more than 1,800 patients with AML. High NADM8 score could efficiently discriminate AML patients with adverse clinical characteristics and genetic lesions and serve as an independent factor predicting a poor prognosis. Immune microenvironment analysis revealed significant enrichment of distinct tumor-infiltrating immune cells and activation of immune checkpoints in patients with high NADM8 scores, acting as a potential biomarker for immune response evaluation in AML. Furthermore, ex vivo drug screening and in vitro experimental validation in a panel of 9 AML cell lines demonstrated that the patients with high NADM8 scores were more sensitive to the PI3K inhibitor, GDC-0914. Finally, functional experiments also substantiated the critical pathogenic role of the SLC25A51 in AML, which could be a promising therapeutic target.ConclusionOur study demonstrated that NAD metabolism-related signature can facilitate risk stratification and prognosis prediction in AML and guide therapeutic decisions including both immunotherapy and targeted therapies.

Abstract Cell-free protein synthesis (CFPS) system is an ideal platform for fast and convenient protein research and has been used for macromolecular assembly, unnatural amino acid embedding, glycoprotein production, and more. To realize the construction of an efficient eukaryotic CFPS platform with the advantages of low cost and short time, a CFPS system based on the yeast Pichia pastoris was built in this study. The internal ribosomal entry site (IRES) can independently initiate translation and thus promote protein synthesis. The Kozak sequences can facilitate translation initiation. Therefore, the screening of IRES and its combination with Kozak was performed, in which cricket paralysis virus (CRPV) exhibited as the best translation initiation element from 14 different IRESs. Furthermore, the system components and reaction environment were explored. The protein yield was nearly doubled by the addition of RNase inhibitor. The cell extract amount, energy regeneration system (phosphocreatine and phosphocreatine kinase), and metal ions (K+ and Mg2+) were optimized to achieve the best protein synthesis yield. This P. pastoris CFPS system can extend the eukaryotic CFPS platform, providing an enabling technology for fast prototyping design and functional protein synthesis. Graphical Abstract

Studies on the mechanism of protein phosphorylation and therapeutic interventions of its related molecular processes are limited by the difficulty in the production of purpose-built phosphoproteins harboring site-specific phosphorylated amino acids or their nonhydrolyzable analogs. Here we address this limitation by customizing the cell-free protein synthesis (CFPS) machinery via chassis strain selection and orthogonal translation system (OTS) reconfiguration screening. The suited chassis strains and reconfigured OTS combinations with high orthogonality were consequently picked out for individualized phosphoprotein synthesis. Specifically, we synthesized the sfGFP protein and MEK1 protein with site-specific phosphoserine (O-pSer) or its nonhydrolyzable analog, 2-amino-4-phosphonobutyric acid (C-pSer). This study successfully realized building cell-free systems for site-specific incorporation of phosphonate mimics into the target protein. Our work lays the foundation for developing a highly expansible CFPS platform and the streamlined production of user-defined phosphoproteins, which can facilitate research on the physiological mechanism and potential interference tools toward protein phosphorylation.

Summary: Esophageal squamous-cell carcinoma (ESCC) is commonly treated with radiotherapy; however, radioresistance hinders its clinical effectiveness, and the underlying mechanism remains elusive. Here, we develop patient-derived xenografts (PDXs) from 19 patients with ESCC to investigate the mechanisms driving radioresistance. Using RNA sequencing, cytokine arrays, and single-cell RNA sequencing, we reveal an enrichment of cancer-associated fibroblast (CAF)-derived collagen type 1 (Col1) and tumor-cell-derived CXCL1 in non-responsive PDXs. Col1 not only promotes radioresistance by augmenting DNA repair capacity but also induces CXCL1 secretion in tumor cells. Additionally, CXCL1 further activates CAFs via the CXCR2-STAT3 pathway, establishing a positive feedback loop. Directly interfering with tumor-cell-derived CXCL1 or inhibiting the CXCL1-CXCR2 pathway effectively restores the radiosensitivity of radioresistant xenografts in vivo. Collectively, our study provides a comprehensive understanding of the molecular mechanisms underlying radioresistance and identifies potential targets to improve the efficacy of radiotherapy for ESCC.

BackgroundWhereas the Montreal Cognitive Assessment (MoCA) and Addenbrooke’s cognitive examination-revised (ACE-R) are commonly used tests for the detection of post-stroke cognitive impairment (PSCI), these instruments take 10–30 min to administer and do not assess processing speed, which is a critical impairment in PSCI. MemTrax (MTx) is a continuous recognition test, which evaluates complex information processing, accuracy, speed, and attention, in 2 min.AimTo evaluate whether MTx is an effective and practical tool for PSCI assessment.MethodsThis study enrolled acute ischemic stroke (AIS) patients who have assessed the cognitive status including MTx, clinical dementia rating (CDR), MoCA, Neuropsychiatric Inventory (NPI), Hamilton depression scale (HAMD), Hamilton anxiety scale (HAMA), the National Institute of Health Stroke Scale (NIHSS), modified Rankin scale (mRS), and Barthel Index of activity of daily living (BI) combined with the physical examinations of the neurologic system at the 90-day (D90) after the AIS. The primary endpoint of this study was establishing MTx cut-offs for distinguishing PSCI from AIS.ResultsOf the 104 participants, 60 were classified to the PSCI group. The optimized cut-off value of MTx-%C (percent correct) was 78%, with a sensitivity and specificity for detecting PSCI from Non-PSCI of 90.0 and 84.1%, respectively, and an AUC of 0.919. Regarding the MTx-Cp (Composite score = MTx-%C/MTx-RT), using 46.3 as a cut-off value, the sensitivity and specificity for detecting PSCI from Non-PSCI were 80.0 and 93.2%, with an AUC of 0.925. Multivariate linear regression showed that PSCI reduced the MTx-%C (Coef. −14.18, 95% CI −18.41∼−9.95, p < 0.001) and prolonged the MTx-RT (response time) (Coef. 0.29, 95% CI 0.16∼0.43, p < 0.001) and reduced the MTx-CP (Coef. −19.11, 95% CI −24.29∼−13.93, p < 0.001).ConclusionMemTrax (MTx) is valid and effective for screening for PSCI among target patients and is a potentially valuable and practical tool in the clinical follow-up, monitoring, and case management of PSCI.

Abstract Background Neoantigens are considered ideal targets for immunotherapy, especially tumor vaccine, because of their strong specificity and immunogenicity. Here, we developed a neoantigen nanovaccine used liposomes with lymph-node targeting characteristic. Methods Our nanovaccine was composed of neoantigens, an amphiphilic liposome and an adjuvant Montanide™ ISA 51. Small animal imaging system and immunofluorescence staining were used to identify the distribution of nanovaccines. A subcutaneous-tumor-resection mouse model of melanoma was established to evaluate the anti-tumor efficacy. Flow cytometry was performed to assay the immune responses initiated by nanovaccines. Results Nanovaccines could traffic to lymph nodes, be uptaken by CD11c+ DCs and promote DCs maturity. After the treatment of our neoantigen nanovaccines, the average recurrence time was extended from 11 to 16 days and the median survival time was even prolonged 7.5 days relative to the control group (NS group). Nanovaccines increased neoantigen-specific T cells to 10-fold of free vaccines, and upregulated Th1 cytokines, such as IFN-γ and TNF-α. The anti-tumor activity of spleen lymphocytes in the nanovaccine group was significantly stronger than that of other groups. However, some immune-inhibitory cells or molecules in tumor microenvironment have been detected upregulated under the immune pressure of neoantigen nanovaccines, such as Tregs and PD-L1. The efficacy of the neoantigen nanovaccine combined with anti-PD1 antibody or Treg inhibiting peptide P60 was better than that of the single treatment. Conclusions We developed a general vaccine strategy, triggering specific T cell responses, and provided feasible combination strategies for better anti-tumor efficacy. Graphical abstract

Abstract Mesenchymal stem cell (MSC)-derived exosomes (Exos) enhanced new bone formation, coupled with positive effects on osteogenesis and angiogenesis. This study aims to define the role of microRNA (miR)-21-5p delivered by human umbilical MSC-derived Exos (hucMSC-Exos) in the osteonecrosis of the femoral head (ONFH). We first validated that miR-21-5p expression was downregulated in the cartilage tissues of ONFH patients. Besides, hucMSCs delivered miR-21-5p to hFOB1.19 cells and human umbilical vein endothelial cells (HUVECs) through the secreted Exos. Loss- and gain-of-function approaches were performed to clarify the effects of Exo-miR-21-5p, SOX5, and EZH2 on HUVEC angiogenesis and hFOB1.19 cell osteogenesis. It was established that Exo-miR-21-5p augments HUVEC angiogenesis and hFOB1.19 cell osteogenesis in vitro, as reflected by elevated alkaline phosphatase (ALP) activity and calcium deposition, and increased the expression of osteogenesis-related markers OCN, Runx2 and Collagen I. Mechanistically, miR-21-5p targeted SOX5 and negatively regulated its expression, while SOX5 subsequently promoted the transcription of EZH2. Ectopically expressed SOX5 or EZH2 could counterweigh the effect of Exo-miR-21-5p. Further, hucMSC-Exos containing miR-21-5p repressed the expression of SOX5 and EZH2 and augmented angiogenesis and osteogenesis in vivo. Altogether, our study uncovered the role of miR-21-5p shuttled by hucMSC-Exos, in promoting angiogenesis and osteogenesis, which may be a potential therapeutic target for ONFH.

Amber suppression has been widely used to incorporate unnatural amino acids (UNAAs) with unique structures or functional side-chain groups into specific sites of the target protein, which expands the scope of protein-coding chemistry. However, this traditional strategy does not allow multiple-site incorporation of different UNAAs into a single protein, which limits the development of unnatural proteins. To address this challenge, the suppression method using multiple termination codons (TAG, TAA or TGA) was proposed, and cell-free unnatural protein synthesis (CFUPS) system was employed. By the analysis of incorporating 3 different UNAAs (p-propargyloxy-l-phenylalanine, p-azyl-phenylalanine and L-4-Iodophenylalanine) and mass spectrometry, the simultaneous usage of the codons TAG and TAA were suggested for better multiple-site UNAA incorporation. The CFUPS conditions were further optimized for better UNAA incorporation efficiency, including the orthogonal translation system (OTS) components, magnesium ions, and the redox environment. This study established a CFUPS approach based on multiple termination codon suppression to achieve efficient and precise incorporation of different types of UNAAs, thereby synthesizing unnatural proteins with novel physicochemical functions.

Objectives and aims: Quantitative MRI (qMRI) has greatly improved the sensitivity and specificity of microstructural brain pathology in multiple sclerosis (MS) when compared to conventional MRI (cMRI). More than cMRI, qMRI also provides means to assess pathology within the normal-appearing and lesion tissue. In this work, we further developed a method providing personalized quantitative T1 (qT1) abnormality maps in individual MS patients by modeling the age dependence of qT1 alterations. In addition, we assessed the relationship between qT1 abnormality maps and patients’ disability, in order to evaluate the potential value of this measurement in clinical practice. Methods: We included 119 MS patients (64 relapsing-remitting MS (RRMS), 34 secondary progressive MS (SPMS), 21 primary progressive MS (PPMS)), and 98 Healthy Controls (HC). All individuals underwent 3T MRI examinations, including Magnetization Prepared 2 Rapid Acquisition Gradient Echoes (MP2RAGE) for qT1 maps and High-Resolution 3D Fluid Attenuated Inversion Recovery (FLAIR) imaging. To calculate personalized qT1 abnormality maps, we compared qT1 in each brain voxel in MS patients to the average qT1 obtained in the same tissue (grey/white matter) and region of interest (ROI) in healthy controls, hereby providing individual voxel-based Z-score maps. The age dependence of qT1 in HC was modeled using linear polynomial regression. We computed the average qT1 Z-scores in white matter lesions (WMLs), normal-appearing white matter (NAWM), cortical grey matter lesions (GMcLs) and normal-appearing cortical grey matter (NAcGM). Lastly, a multiple linear regression (MLR) model with the backward selection including age, sex, disease duration, phenotype, lesion number, lesion volume and average Z-score (NAWM/NAcGM/WMLs/GMcLs) was used to assess the relationship between qT1 measures and clinical disability (evaluated with EDSS). Results: The average qT1 Z-score was higher in WMLs than in NAWM. (WMLs: 1.366 ± 0.409, NAWM: −0.133 ± 0.288, [mean ± SD], p

Site-specific incorporation of unnatural amino acids (UNAAs) into proteins using an orthogonal translation system (OTS) has expanded the scope of protein-coding chemistry. The key factor affecting UNAA embedding efficiency is the orthogonality of the OTS. Compared to traditional cell systems, cell-free systems are more convenient to control the reaction process and improve the utilization rate of UNAA. In this study, a linear DNA template-based cell-free unnatural protein synthesis system for rapid high-throughput screening and evolution was proposed. A total of 14 cell extracts were selected for screening out cell extract with high expression level. The result showed that EcAR7 ΔA ΔSer cell extract was optimal for the cell-free system. In addition, the screening results of four UNAAs, p-propargyloxy-l-phenylalanine (pPaF), p-azyl-phenylalanine (pAzF), p-acetyl-l-phenylalanine (pAcF), and p-benzoyl-l-phenylalanine (pBpF), showed that o-aaRS and o-tRNA of pPaF had good orthogonality. A new pair of corresponding o-aaRS and o-tRNA for pBpF was screened out. These results proved that this method could speed up the screening of optimal OTS components for UNAAs with versatile functions.

BackgroundThe risk factors for acute heart failure (AHF) vary, reducing the accuracy and convenience of AHF prediction. The most common causes of AHF are coronary heart disease (CHD). A short-term clinical predictive model is needed to predict the outcome of AHF, which can help guide early therapeutic intervention. This study aimed to develop a clinical predictive model for 1-year prognosis in CHD patients combined with AHF.Materials and methodsA retrospective analysis was performed on data of 692 patients CHD combined with AHF admitted between January 2020 and December 2020 at a single center. After systemic treatment, patients were discharged and followed up for 1-year for major adverse cardiovascular events (MACE). The clinical characteristics of all patients were collected. Patients were randomly divided into the training (n = 484) and validation cohort (n = 208). Step-wise regression using the Akaike information criterion was performed to select predictors associated with 1-year MACE prognosis. A clinical predictive model was constructed based on the selected predictors. The predictive performance and discriminative ability of the predictive model were determined using the area under the curve, calibration curve, and clinical usefulness.ResultsOn step-wise regression analysis of the training cohort, predictors for MACE of CHD patients combined with AHF were diabetes, NYHA ≥ 3, HF history, Hcy, Lp-PLA2, and NT-proBNP, which were incorporated into the predictive model. The AUC of the predictive model was 0.847 [95% confidence interval (CI): 0.811–0.882] in the training cohort and 0.839 (95% CI: 0.780–0.893) in the validation cohort. The calibration curve indicated good agreement between prediction by nomogram and actual observation. Decision curve analysis showed that the nomogram was clinically useful.ConclusionThe proposed clinical prediction model we have established is effective, which can accurately predict the occurrence of early MACE in CHD patients combined with AHF.

ObjectiveAs an important biomarker to reflect tumor cell proliferation and tumor aggressiveness, Ki-67 is closely related to the high early recurrence rate and poor prognosis, and pretreatment evaluation of Ki-67 expression possibly provides a more accurate prognosis assessment and more better treatment plan. We aimed to develop a nomogram based on gadolinium ethoxybenzyl diethylenetriamine pentaacetic acid (Gd-EOB-DTPA)-enhanced magnetic resonance imaging (MRI) combined with T1 mapping to predict Ki-67 expression in hepatocellular carcinoma (HCC).MethodsThis two-center study retrospectively enrolled 148 consecutive patients who underwent preoperative Gd-EOB-DTPA-enhanced MRI T1 mapping and surgically confirmed HCC from July 2019 to December 2020. The correlation between quantitative parameters from T1 mapping, ADC, and Ki-67 was explored. Three cohorts were constructed: a training cohort (n = 73) and an internal validation cohort (n = 31) from Shunde Hospital of Southern Medical University, and an external validation cohort (n = 44) from the Sixth Affiliated Hospital, South China University of Technology. The clinical variables and MRI qualitative and quantitative parameters associational with Ki-67 expression were analyzed by univariate and multivariate logistic regression analyses. A nomogram was developed based on these associated with Ki-67 expression in the training cohort and validated in the internal and external validation cohorts.ResultsT1rt-Pre and T1rt-20min were strongly positively correlated with Ki-67 (r = 0.627, r = 0.607, P < 0.001); the apparent diffusion coefficient value was moderately negatively correlated with Ki-67 (r = -0.401, P < 0.001). Predictors of Ki-67 expression included in the nomogram were peritumoral enhancement, peritumoral hypointensity, T1rt-20min, and tumor margin, while arterial phase hyperenhancement (APHE) was not a significant predictor even included in the regression model. The nomograms achieved good concordance indices in predicting Ki-67 expression in the training and two validation cohorts (0.919, 0.925, 0.850), respectively.ConclusionsT1rt-Pre and T1rt-20min had a strong positive correlation with the Ki-67 expression in HCC, and Gd-EOB-DTPA enhanced MRI combined with T1 mapping-based nomogram effectively predicts high Ki-67 expression in HCC.

Abstract Radiotherapy remains the mainstay for treatment of various types of human cancer; however, the clinical efficacy is often limited by radioresistance, in which the underlying mechanism is largely unknown. Here, using esophageal squamous cell carcinoma (ESCC) as a model, we demonstrate that guanine nucleotide exchange factor 2 (VAV2), which is overexpressed in most human cancers, plays an important role in primary and secondary radioresistance. We have discovered for the first time that VAV2 is required for the Ku70/Ku80 complex formation and participates in non-homologous end joining repair of DNA damages caused by ionizing radiation. We show that VAV2 overexpression substantially upregulates signal transducer and activator of transcription 1 (STAT1) and the STAT1 inhibitor Fludarabine can significantly promote the sensitivity of radioresistant patient-derived ESCC xenografts in vivo in mice to radiotherapy. These results shed new light on the mechanism of cancer radioresistance, which may be important for improving clinical radiotherapy.

BackgroundPatients with diabetes have an increased risk of developing vulnerable plaques (VPs), in which dyslipidemia and chronic inflammation play important roles. Non-high-density lipoprotein cholesterol (non-HDL-C) and neutrophil-lymphocyte ratio (NLR) have emerged as potential markers of both coronary artery VPs and cardiovascular prognosis. This study aimed to investigate the predictive value of non-HDL-C and NLR for coronary artery VPs in patients with type 2 diabetes mellitus (T2DM).MethodsWe retrospectively enrolled 204 patients with T2DM who underwent coronary computed tomography angiography between January 2018 and June 2020. Clinical data including age, sex, hypertension, smoking, total cholesterol, low-density lipoprotein cholesterol, HDL-C, triglyceride, non-HDL-C, glycated hemoglobin, neutrophil count, lymphocyte count, NLR, and platelet count were analyzed. Multivariate logistic regression was used to estimate the association between non-HDL-C, NLR, and coronary artery VPs. Receiver operating curve analysis was performed to evaluate the value of non-HDL-C, NLR, and their combination in predicting coronary artery VPs.ResultsIn our study, 67 patients (32.84%) were diagnosed with VPs, 75 (36.77%) with non-VP, and 62 (30.39%) with no plaque. Non-HDL-C and NLR were independent risk factors for coronary artery VPs in patients with T2DM. The areas under the ROC curve of non-HDL-C, NLR, and their combination were 0.748 [95% confidence interval (CI): 0.676–0.818], 0.729 (95% CI: 0.650–0.800), and 0.825 (95% CI: 0.757–0.887), respectively.ConclusionEither non-HDL-C or NLR could be used as a predictor of coronary artery VPs in patients with T2DM, but the predictive efficiency and sensitivity of their combination would be better.

Abstract Background Monacolin J (MJ) is a key intermediate for the synthesis of cholesterol-lowering drug simvastatin. Current industrial production of MJ involves complicated chemical hydrolysis of microbial fermented lovastatin. Recently, heterologous production of MJ has been achieved in yeast and bacteria, but the resulting metabolic stress and excessive accumulation of the compound adversely affect cell activity. Results Five genes, tapA, stapA, slovI, smokI and smlcE, coding for fungal statin pump proteins were expressed in an MJ producing yeast strain, Komagataella phaffii J#9. Overexpression of these genes facilitated MJ production. Among them, tapA from Aspergillus terreus highly improved MJ production and led to a titer increase of 108%. Exogenous MJ feeding study on an MJ non-producing strain GS-P GAP -TapA was then performed, and the results illustrated tough entry of MJ into cells and possible efflux action of TapA. Further, intracellular and extracellular MJ levels of J#9 and J#9-TapA were analyzed. The extracellular MJ level of J#9-TapA increased faster, but its intracellular MJ percentage kept lower as compared to J#9. The results proved that TapA effectively excreted MJ from cells. Then functions of TapA were evaluated in a high-production bioreactor fermentation. Differently, TapA expression caused a low MJ titer but high intracellular MJ accumulation in J#9-TapA compared with J#9. Conclusions Statin pump proteins improved MJ production in K. phaffii in a shake flask. Exogenous MJ feeding and endogenous MJ producing experiments demonstrated the efflux function of TapA. TapA improved MJ production at low MJ levels in a shake flask, but decreased it at high MJ levels in a bioreactor. This finding is useful for statin pump improvement and metabolic engineering for statin bioproduction.

BackgroundThe dementia and affective disorders are common non-motor features in patients with essential tremor (ET). However, the relationship of ET with cognitive impairments and affective disorders remains controversial. This meta-analysis aimed to analyze the association of ET with dementia and affective disorders.MethodsOriginal studies published from January 1999 to October 2019 were systematically searched from the database of Medline (OvidSP), EMBASE (OvidSP), and the Cochrane Central Register of Controlled Trials. Pooled standard mean difference (SMD, random effect model), odds ratios (ORs), relative risk (RR), and 95% CI were calculated.ResultsCompared with the Non-ET group, patients with ET had significantly lower Mini-Mental State Examination (MMSE) score (SMD, −1.16; 95% CI, −1.75 to −0.58; p = 0.0001) and had significantly higher depressive and anxiety symptoms scale score (SMD, 0.55; 95% CI, 0.22–0.87; p = 0.0009). The OR for dementia and affective disorders in individuals with ET compared with individuals without ET was 2.49 (95% CI, 2.17–2.85, p < 0.00001). While there was no significant difference in Montreal Cognitive Assessment (MoCA) score between ET and Non-ET groups (SMD, −0.52; 95% CI, −0.16 to 0.13; p = 0.23), there was a significant difference in the risk of mortality between ET and Non-ET groups (RR = 4.69, 95% CI, 2.18–10.07).ConclusionThe non-motor symptoms should not be neglected among patients with ET. However, the causal relationship between ET and dementia, depression, and anxiety is unclear.

ObjectiveTo evaluate and compare the predictive value of Face, Arm, Speech Test (FAST) and Balance, Eyes, Face, Arm, Speech, Time (BEFAST) scale in the acute ischemic stroke (AIS).MethodsWe searched Medline and Ovid databases for relevant literature in the English language. There were no limitations on the date. The sensitivity, specificity, likelihood ratio, and diagnostic odds ratio were pooled for meta-analysis. The symmetric receiver operator characteristic curve and Fagan's Nomogram were drawn, and meta-regression and subgroup analysis were used to explore the source of heterogeneity.ResultsA total of 9 studies, including 6,151 participants, were analyzed. The combined sensitivity of FAST was 0.77 [95% CI (0.64–0.86)], specificity was 0.60 [95% CI (0.38–0.78)], the area under the ROC curve was 0.76, and the diagnostic ratio was 1.57, while the sensitivity of BEFAST was 0.68 [95% CI (0.23–0.93)], specificity was 0.85 [95% CI (0.72–0.92)], the area under the ROC curve was 0.86, and the diagnostic odds ratio was 2.44. No publication bias was detected in Deeks' funnel plot. For FAST, meta-regression analysis showed that the prospective design, satisfactory description of the index test, and a broad spectrum of disease contributed to the heterogeneity in sensitivity, while no sources contributed to the heterogeneity in sensitivity. When the pretest probability was set as 20%, the posterior probability in Fagan's Nomogram was 32%; however, when the pretest probability was set as 20% in BEFAST, the posterior probability in Fagan's Nomogram was 52%.ConclusionsOur findings indicated that FAST and BEFAST might be useful in the diagnosis of acute ischemic stroke. The diagnostic value of BEFAST in acute ischemic stroke was higher than in FAST; thus, it might have an important role in the fast recognition of acute ischemic stroke.

The cell-free protein synthesis (CFPS) system, as a technical core of synthetic biology, can simulate the transcription and translation process in an in vitro open environment without a complete living cell. It has been widely used in basic and applied research fields because of its advanced engineering features in flexibility and controllability. Compared to a typical crude extract-based CFPS system, due to defined and customizable components and lacking protein-degrading enzymes, the protein synthesis using recombinant elements (PURE) system draws great attention. This review first discusses the elemental composition of the PURE system. Then, the design and preparation of functional proteins for the PURE system, especially the critical ribosome, were examined. Furthermore, we trace the evolving development of the PURE system in versatile areas, including prototyping, synthesis of unnatural proteins, peptides and complex proteins, and biosensors. Finally, as a state-of-the-art engineering strategy, this review analyzes the opportunities and challenges faced by the PURE system in future scientific research and diverse applications.

Science education is attracting increasing attention and many researchers focus on the issue about the attitude-achievement relationship in science, but there is still no consistent conclusion. By using a three-level meta-analytic approach, the aim of the current study was to investigate the relationship between attitude toward science and academic achievement in learning science among primary and secondary school students, and to explore if some study characteristics could have contributed to the inconsistent findings with regard to this relationship as observed in the research literature. A total of 37 studies with 132 effect sizes involving a total of 1,042,537 participants were identified. The meta-analytic results revealed that there was an overall positive and moderate relationship between attitude toward science and learning achievement in science (r = 0.248, p < 0.001). The results further found that this association was moderated by the type of attitude and larger effect sizes were shown in self-efficacy than in interest, societal relevance of attitude toward science, and mixed attitude. Moreover, the effect sizes of studies with unstandardized measure to assess science achievement were larger than those with standardized measure. Possible explanations for these findings and its implications for future research directions were also discussed in this review.

With the increasing demand of human beings for space exploration, space robots show great development potential. When grasping space objects with different sizes and shapes, cable-driven continuum arms have better performance than traditional robots. In this paper, a novel space robot with triple cable-driven continuum arms is proposed, which can achieve compliant grasping through multi-arm cooperation. The kinematic model of the robot is proposed and verified through simulations and experiments. Results show that the maximum repeat positioning error is no larger than 1 mm and the maximum tracking error is no larger than 2 mm, compared to the 300 mm long arm. In addition, the demonstration experiment of grasping a ball indicates the good performance of the robot in compliant grasping.

Circular RNA (circRNA) is a unique type of noncoding RNA molecule. Compared with traditional linear RNA, circRNA is a covalently closed circle produced by a process called backsplicing. CircRNA is abundant in many cells and has rich functions in cells, such as acting as miRNA sponge, protein sponge, protein scaffold, and mRNA regulator. With the continuous development of circRNA study, circRNA has also played an important role in medical applications, including circRNA vaccines and gene therapy. In this review, we illustrate the synthesis of circRNAs in vitro. We focus on biological ligation methods, such as enzymatic ligation from the bacteriophage T4 and ribozyme method. In addition, we summarize the current challenges in the design, synthesis, application, and production of circRNAs, and propose possible solutions in the future. CircRNA is expected to play an essential role in basic research and medical applications.

Cell-free gene expression systems with linear DNA expression templates (LDETs) have been widely applied in artificial cells, biochips, and high-throughput screening. However, due to the degradation caused by native nucleases in cell extracts, the transcription with linear DNA templates is weak, thereby resulting in low protein expression level, which greatly limits the development of cell-free systems using linear DNA templates. In this study, the protective sequences for stabilizing linear DNA and the transcribed mRNAs were rationally designed according to nucleases’ action mechanism, whose effectiveness was evaluated through computer simulation and cell-free gene expression. The cell-free experiment results indicated that, with the combined protection of designed sequence and GamS protein, the protein expression of LDET-based cell-free systems could reach the same level as plasmid-based cell-free systems. This study would potentially promote the development of the LDET-based cell-free gene expression system for broader applications.

With the rapid growth of the economy and population, the desire to use groundwater supplies is increasing. In order to detect groundwater, conducting a geophysics survey is a common way to map the subsurface. This method can describe the subsurface’s physical properties. For example, the direct current (DC) resistivity method provides a resistivity map of the subsurface, in which the groundwater is normally located in low resistivity anomaly zones. However, the DC method needs to overcome the challenges of the urban environment, with its infrastructure and background noise. In this paper, we propose a pseudo-3D resistivity surveying method to solve these problems, and it is applied to situations of groundwater detection within the city environment, dam leakage, and drinking water. From the pseudo-3D resistivity surveying, we detected the low-resistivity anomaly in the area, which is interpreted as groundwater through borehole verification.

The present study is the first to empirically test a hierarchical, positive-oriented model of the self and its relationship to second language (L2) achievement motivation, and compare it in three different cultural contexts of Japan, the United States, and Taiwan. Based on the L2 self-model (Lake, 2016), three levels of constructs were developed: Global Self (i.e., Flourishing, Curiosity, and Hope); Positive L2 domain self (i.e., interested-in-L2 self, harmonious passion for L2 learning, and mastery L2 goal orientation); and L2 Motivational Variables (i.e., reading, speaking and listening self-efficacy). A total of 667 students participated in this study, including 181 first-year college students in Japan, 159 high school students in Taiwan, and 327 community college students in the United States. All the participants were learning L2 in school. Results showed that the measures of positive global self, L2 domain self, and L2 motivational self all had a stronger relationship within their respective levels, and progressively weaker relationships as level of generality/specificity became more distal. Furthermore, the relationships among measures varied in the differing cultural contexts with the Japan-based student participants relatively lower on all measures. Implications for teacher educators in the L2 context have been discussed.

Objective: To investigate changes in brain function at the regional and whole-brain levels in patients with tremor-dominant Parkinson’s disease (TDPD) complicated by sleep disorder (SD) by regional homogeneity (ReHo) and functional connectivity (FC) analysis of whole-brain resting-state functional magnetic resonance images.Materials and Methods: ReHo and seed-based FC analyses were conducted among 32 patients with TDPD and SD (TDPD-SD), 24 with TDPD and no SD (TDPD-NSD), and 23 healthy controls (HCs) to assess spontaneous brain activity and network-level brain function. Correlation analyses were used to examine the associations between brain activity and the clinical data.Results: Anterior cingulate gyrus (ACC) ReHo values differed significantly among the groups. ACC ReHo values were increased in TDPD-SD vs. HC and TDPD-SD vs. TDPD-NSD. ACC ReHo values were reduced in TDPD-NSD vs. HC. TDPD-SD ReHo values were positively correlated with Pittsburgh Sleep Quality Index (PSQI) scores (r = 0.41, p = 0.020) but negatively correlated with Parkinson’s Disease Sleep Scale (PDSS) scores (r = −0.38, p = 0.030). FC analysis using ACC as a mask showed that FC of the left olfactory cortex (L-OC), right straight gyrus (R-SG), right superior parietal gyrus (R-SPG), and right precuneus differed significantly among the groups. FC values between R-SG and ACC were significantly lower in TDPD-SD than in TDPD-NSD, while the FC of L-OC and R-OC with ACC was significantly lower in TDPD-SD than in HC. FC between ACC and L-OC, R-SPG, and the right precuneus was lower in TDPD-NSD than in HC. There was no correlation between the FC values and other clinical data in any of the groups.Conclusion: Localized abnormal activity in TDPD-SD was chiefly triggered by ACC. The change in the ReHo of ACC is closely related to the severity of TDPD-associated SD, revealing the role of this region as a regulator of the sleep mechanism in TDPD. Significant abnormal FC was found between R-SG and ACC in TDPD-SD but was not shown to correlate with clinical data.

The present contribution offers an overview of a new area of research in the field of foreign language acquisition, which was triggered by the introduction of Positive Psychology (PP) (MacIntyre and Gregersen, 2012). For many years, a cognitive perspective had dominated research in applied linguistics. Around the turn of the millennium researchers became increasingly interested in the role of emotions in foreign language learning and teaching, beyond established concepts like foreign language anxiety and constructs like motivation and attitudes toward the foreign language. As a result, a more nuanced understanding of the role of positive and negative learner and teacher emotions emerged, underpinned by solid empirical research using a wide range of epistemological and methodological approaches. PP interventions have been carried out in schools and universities to strengthen learners and teachers’ experiences of flow, hope, courage, well-being, optimism, creativity, happiness, grit, resilience, strengths, and laughter with the aim of enhancing learners’ linguistic progress. This paper distinguishes the early period in the field that started with MacIntyre and Gregersen (2012), like a snowdrop after winter, and that was followed by a number of early studies in relatively peripheral journals. We argue that 2016 is the starting point of the current period, characterized by gradual recognition in applied linguistics, growing popularity of PP, and an exponential increase in publications in more mainstream journals. This second period could be compared to a luxuriant English garden in full bloom.

Are bilingualism and/or biculturalism good for a person’s positive well being? A growing number of studies have shown different positive outcomes of being exposed to two cultures or speaking two languages respectively, but the benefits of being both bilingual and bicultural have rarely been investigated theoretically or empirically. The purpose of this paper is to summarize the main beneficial outcomes of bilingualism and biculturalism, and to integrate these benefits into a new conceptual framework: Positive Bilingualism and Biculturalism GEAR model. The GEAR model suggests that the beneficial outcomes of bilingualism and biculturalism can be systematically classified into four positive dimensions (psychological Growth; cognitive Exploration; linguistic Awareness; and social Reinforcement), in which there are dynamic interactions among these four dimensions. The hypothetical GEAR model provides an intricate theoretical approach to understand the potential benefits to an individual of experiencing more than one language and one culture in their life. The proposed model in this research offers a systematic framework for conducting future research to examine whether bilingualism and biculturalism accrue benefits to the individual.

This study aimed to explore the cognitive and affective factors that predict the Psychological capital (PsyCap) of Chinese primary school teachers. The participants were 1,384 teachers from 34 primary schools in Chengdu city, in southwestern part of China. A resource model of PsyCap was proposed and tested using structural equation modeling (SEM). The model was tested with two independent variables of growth mindset and well-being as predictors of PsyCap. Results showed that both cognitive construct (i.e., growth mindset) and affective construct (i.e., well-being) were positively and significantly related to the PsyCap. In addition, SEM results indicated a significant influence of both growth mindset and well-being on all the variables of PsyCap (i.e., Efficacy, Hope, Optimism, and Resilience). Explanations and implications were discussed for the findings, and some limitations were also discussed.

The current study investigated the relationships among growth mindset, work engagement, perseverance of effort and well-being for secondary school teachers in the Chinese educational context. We adopted Growth Mindset Inventory, Utrecht Work Engagement Scale (UWES), Grit Scale (Perseverance subscale), and PERMA profiles that measure five dimensions of well-being. Participants included 472 secondary school teachers from 10 secondary schools in central China. Correlation analysis showed that growth mindset, well-being, and perseverance of effort could all predict work engagement. Moreover, the structural equation model and mediation analysis further suggested that well-being and perseverance of effort could partially mediate the relationship between growth mindset and work engagement. This study permitted to advance our knowledge about the relationship between growth mindset and work engagement, which should be considered for future teaching practices and teacher development.

The scarcity of primordial germ cells (PGCs) in the developing mammalian embryo hampers robust biochemical analysis of the processes that underlie early germ cell formation. Here, we demonstrate that DAZL, a germ cell-specific RNA binding protein, is a robust PGC marker during in vitro germ cell development. Using Dazl-GFP reporter ESCs, we demonstrate that DAZL plays a central role in a large mRNA/protein interactive network that blocks the translation of core pluripotency factors, including Sox2 and Sall4, as well as of Suz12, a polycomb family member required for differentiation of pluripotent cells. Thus, DAZL limits both pluripotency and somatic differentiation in nascent PGCs. In addition, we observed that DAZL associates with mRNAs of key Caspases and similarly inhibits their translation. This elegant fail-safe mechanism ensures that, whereas loss of DAZL results in prolonged expression of pluripotency factors, teratoma formation is avoided due to the concomitant activation of the apoptotic cascade.

Using a positive psychological framework, Broaden-and-Build Theory, we investigated whether and how positive and negative emotions (affective factors) and cognitive flexibility are related to flourishing. Participants are 151 bilingual adults from diverse backgrounds enrolled in a local community college. Results showed that both positive and negative emotions and cognitive flexibility were significantly related to flourishing. More importantly, mediation effects from emotions to flourishing through cognitive flexibility occurred only from positive emotions. These findings are the first to explore how affective factors are related to flourishing in a sample of bilingual adults residing in a predominantly monolingual country. Practical implications are discussed with regards to the cultivation of positive emotions and fostering cognitive flexibility among bilinguals in and out of school.

Background: Microsatellite instability (MSI) is associated with treatment response and prognosis in patients with rectal cancer (RC). However, intratumoral heterogeneity limits MSI testing in patients with RC. The authors developed a subregion radiomics model based on multiparametric MRI to preoperatively assess high-risk subregions with MSI and predict the MSI status of patients with RC. Methods: This retrospective study included 475 patients (training cohort, 382; external test cohort, 93) with RC from two participating hospitals between April 2017 and June 2023. In the training cohort, subregion radiomic features were extracted from multiparametric MRI, which included T2-weighted, T1-weighted, diffusion-weighted, and contrast-enhanced T1-weighted imaging. MSI-related subregion radiomic features, classical radiomic features, and clinicoradiological variables were gathered to build five predictive models using logistic regression. Kaplan-Meier survival analysis was conducted to explore the prognostic information. Results: Among the 475 patients [median age, 64 years (interquartile range, IQR: 55-70 years); 304 men and 171 women], the prevalence of MSI was 11.16% (53/475). The subregion radiomics model outperformed the classical radiomics and clinicoradiological models in both training [area under the curve (AUC) =0.86, 0.72, and 0.59, respectively] and external test cohorts (AUC=0.83, 0.73, and 0.62, respectively). The subregion-clinicoradiological model combining clinicoradiological variables and subregion radiomic features performed the optimal, with AUCs of 0.87 and 0.85 in the training and external test cohorts, respectively. The 3-year disease-free survival rate of MSI groups predicted based on the model was higher than that of the predicted microsatellite stability groups in both patient cohorts (training, P=0.032; external test, P= 0.046). Conclusions: The authors developed and validated a model based on subregion radiomic features of multiparametric MRI to evaluate high-risk subregions with MSI and predict the MSI status of RC preoperatively, which may assist in individualized treatment decisions and positioning for biopsy. [ABSTRACT FROM AUTHOR]