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Obesity and overweight conditions have become major health challenges worldwide. The exploration of effective weight loss strategies is essential. Nudges are currently advancing approaches that represent a new and better method for changing the behaviors of people. However, the effectiveness of nudge interventions on weight loss in overweight people who may be obese has not been synthesized in a systematic manner. In this study, a systematic literature search was performed. Only randomized controlled trials (RCTs) were considered. Weighted mean differences (WMDs) and 95% confidence intervals (CIs) were calculated as summary statistics. In total, 25 RCTs involving a population of 5,929 individuals were included. Significant effects of the nudge strategy on weight loss (WMD: -0.96 kg, 95% CI: -1.49 to -0.43), body mass index (WMD: -0.3 kg/m2, 95% CI: -0.41 to -0.19) and waist circumference (WMD: -0.75 cm, 95% CI: -1.23 to -0.27) were observed. The subgroup analysis showed that the reduction in body weight associated with nudge interventions was significant in younger and more obese people. Moreover, the effect of nudge intervention on weight loss weakened over time. Overall, the nudge strategy can promote changes in weight loss, body mass index and waist circumference of adults, albeit at a mild magnitude and in particular types of individuals. Nudge strategies can be recommended to clinical practitioners and policy-makers to promote obesity management.

The funny current, I f, was first recorded in the heart 40 or more years ago by Dario DiFrancesco and others. Since then, we have learnt that I f plays an important role in pacemaking in the sinus node, the innate pacemaker of the heart, and more recently evidence has accumulated to show that I f may play an important role in action potential conduction through the atrioventricular (AV) node. Evidence has also accumulated to show that regulation of the transcription and translation of the underlying Hcn genes plays an important role in the regulation of sinus node pacemaking and AV node conduction under normal physiological conditions - in athletes, during the circadian rhythm, in pregnancy, and during postnatal development - as well as pathological states - ageing, heart failure, pulmonary hypertension, diabetes and atrial fibrillation. There may be yet more pathological conditions involving changes in the expression of the Hcn genes. Here, we review the role of I f and the underlying HCN channels in physiological and pathological changes of the sinus and AV nodes and we begin to explore the signalling pathways (microRNAs, transcription factors, GIRK4, the autonomic nervous system and inflammation) involved in this regulation. This review is dedicated to Dario DiFrancesco on his retirement.

Gut microbial dysbiosis has been linked to many noncommunicable diseases. However, little is known about specific gut microbiota composition and its correlated metabolites associated with molecular signatures underlying host response to infection. Here, we describe the construction of a proteomic risk score based on 20 blood proteomic biomarkers, which have recently been identified as molecular signatures predicting the progression of the COVID-19. We demonstrate that in our cohort of 990 healthy individuals without infection, this proteomic risk score is positively associated with proinflammatory cytokines mainly among older, but not younger, individuals. We further discover that a core set of gut microbiota can accurately predict the above proteomic biomarkers among 301 individuals using a machine learning model and that these gut microbiota features are highly correlated with proinflammatory cytokines in another independent set of 366 individuals. Fecal metabolomics analysis suggests potential amino acid-related pathways linking gut microbiota to host metabolism and inflammation. Overall, our multi-omics analyses suggest that gut microbiota composition and function are closely related to inflammation and molecular signatures of host response to infection among healthy individuals. These results may provide novel insights into the cross-talk between gut microbiota and host immune system.

Cordycepin is a major bioactive compound found in Cordyceps sinensis that exhibits a broad spectrum of biological activities. Here a Paecilomyces hepiali OR-1 strain was initially isolated from plateau soil for the bioproduction of cordycepin. Subsequently, strain modification including 60Co γ-ray and ultraviolet irradiation were employed to increase the cordycepin titer, resulted in a high-yield mutant strain P. hepiali ZJB18001 with the cordycepin content of 0.61 mg/gDCW, showing a 2.3-fold to that from the wild strain (0.26 mg/gDCW). Furthermore, medium screening based on Box-Behnken design and the response surface methodology facilitated the enhancement of cordycepin yield to the value of 0.96 mg/gDCW at 25 °C for 5 days in submerged cultivation with an optimized medium composition. The high cordycepin yield, rapid growth rate and stable genetic characteristics of P. hepiali ZJB18001 are beneficial in terms of costs and time for the industrialization of cordycepin production.

Metastasized cancer cells have an increased resistance to therapies leading to a drastic decrease in patient survival rates. However, our understanding of the cause for this enhanced resistance is lacking. In this study, we report that physically tight confinement during cancer cell migration triggers therapeutic resistance and induces cancer stem cell-like behavior including up-regulation in efflux proteins and in cancer stem cell related markers. Moreover, the re-localization of Yes-associated protein (YAP) to the cell nucleus indicated an elevated level of cytoskeletal tension. The increased cytoskeletal tension suggested that mechanical interactions between cancer cells and tight surroundings during metastasis is one of the factors that contributes to therapeutic resistance and acquisition of cancer stem cell (CSC) like features. With this system and supporting data, we are able to study cells with therapeutic resistance and CSC-like properties for the future purpose of developing new strategies for the treatment of metastatic cancer.

Aims Current physical activity guidelines emphasize little on light-intensity physical activity (LPA) in terms of reducing the risk of cardiovascular mortality. This meta-analysis aimed to bridge this gap by assessing their association using objectively measured LPA data. Methods and results Databases of PubMed and Scopus were searched to April 2020 for prospective cohort studies that reported the association of LPA assessed by activity monitors with the risk of cardiovascular mortality in the general population. Multivariable-adjusted hazard ratios (HRs) with 95% confidence intervals (CIs) were pooled using a random-effects model. Dose–response and subgroup analyses were also performed. Six cohort studies with seven datasets enrolling 13 960 participants were included. LPA was all measured by accelerometers. The HR of LPA per 30 min/day for cardiovascular mortality was pooled to be 0.80 (95% CI 0.67–0.96). This association was non-linearly shaped (Pnonlinearity < 0.01) and unaffected by sex difference. Moreover, substituting LPA for sedentary time of 30 min/day lowered the risk of cardiovascular mortality by 16% (95% CI 0.73–0.96). Results showed further that LPA was inferior to moderate-to-vigorous physical activity in reducing the risk of cardiovascular mortality when performed with an equal time-length set at 30 min/day (HR 0.83 vs. 0.54, Pcomparison = 0.046), but became comparable if at an equal activity-amount set at 150 metabolic equivalents-min/day (HR 0.67 vs. 0.54, Pcomparison = 0.41). Conclusion LPA shows potential in reducing the risk of cardiovascular mortality, and interventions targeting at LPA improvement are worth being encouraged.

Here, ferric oxide-loaded metal-organic framework (FeTCPP/Fe2O3 MOF) nanorice was designed and constructed by the liquid diffusion method. The introduction of iron metal nodes and the loading of Fe2O3 can effectively catalyze the Fenton reaction to produce hydroxyl radicals (•OH) and overcome the hypoxic environment of tumor tissue by generating oxygen. The monodispersity and porosity of the porphyrin photosensitizers in the MOF structure exposed more active sites, which promoted energy exchange between porphyrin molecules and oxygen molecules for photodynamic therapy (PDT) treatment. Therefore, the generated hydroxyl radicals and singlet oxygen (1O2) can synergistically act on tumor cells to achieve the purpose of improving tumor therapy. Then the erythrocyte membrane was camouflaged to enhance blood circulation and tissue residence time in the body, and finally, the targeted molecule AS1411 aptamer was modified to achieve the high enrichment of MOF photosensitizers on a tumor domain. As a result, the MOF nanorice camouflaged by the erythrocyte membrane can effectively reduce side effects and improve the therapeutic effect of PDT and chemo-dynamic therapy (CDT). The study not only improved the efficacy of PDT and CDT in essence from the MOF nanorice but also used the camouflage method to further concentrate FeTCPP/Fe2O3 on the tumor sites, achieving the goal of multiple gains. These results will provide theoretical and practical directions for the development of tumor-targeted MOF nanomaterials.

Xylitol is a widely used natural sweetener for the reduction of excessive sugar consumption. However, concerns of xylitol consumption existed as it is a highly permeable substance in the colon that could cause diarrhea and other adverse symptoms. To assess the relationship between xylitol dosage and diarrhea, especially the influences of diarrhea on physiological characteristics, the immune system, and gut microbiota in rats, the control, low-dose (L), medium-dose (M), and high-dose (H) groups were fed with 0, 1, 3, and 10% of xylitol, respectively, correspondingly for 15 days, followed by a 7-day recovery. Only medium- and high-dose xylitol would cause diarrhea in rats. Quantitative imaging of colonic tissue and the expression levels of proinflammatory factors revealed a higher degree of immune responses in the rats from H groups but statistically stable in M groups, despite that light diarrhea was observed. A shift of the gut microbiota composition was observed in the rats from H groups, including significant decreases of genera Ruminococcaceae and Prevotella and a notable increase and colonization of Bacteroides, accompanied with changes of short-chain fatty acid production. Tolerance and adaptation to xylitol consumption were observed in a dose-dependent manner. Our findings demonstrate that diarrhea caused by the high dosage of xylitol can exert distinctive changes on gut microbiota and lay the foundation to explore the mechanism underlying the shift in gut microbiota composition.

Creatinine-to-cystatin C ratio has attracted substantial interest as a measure to reflect health well-being, but no studies have assessed whether its longitudinal changes are associated with risk of diabetes. We aimed to examine their association, along with the exploration of the relationship of such changes with cardiometabolic control in middle-aged and older adults.We included a total of 3278 participants aged ≥45 years who provided measurements of creatinine and cystatin C at baseline and 4 years later from the China Health and Retirement Longitudinal Study. Diabetes was diagnosed based on glucose, hemoglobin A1c (HbA1c), medical history, or use of antidiabetic mediations. Odds ratio (OR) and 95% confidence interval (CI) were obtained using logistic regression analyses.After 4-year follow-up, 272 participants developed diabetes. Larger increases in creatinine-to-cystatin C ratio were associated with lower risk of diabetes. The multivariable-adjusted OR for diabetes per 1 SD increase in creatinine-to-cystatin C ratio was 0.84 (95% CI 0.72-0.98). Compared with participants showing decreases in creatinine-to-cystatin C ratio but increases in body mass index (BMI), those experiencing increases in creatinine-to-cystatin C ratio and decreases in BMI had the largest risk reduction (multivariable-adjusted OR 0.52). Changes in creatinine-to-cystatin C ratio showed inverse correlation with blood pressure, HbA1c, lipids, and C-reactive protein at the 4-year follow-up. Moreover, they also correlated inversely with changes in HbA1c and C-reactive protein (all P ≤ 0.004).Increases in creatinine-to-cystatin C ratio led to reduced risk of diabetes and may benefit cardiometabolic control.背景: 肌酐与胱抑素C比值作为反映健康状况的一种指标获得了广泛关注, 但没有研究评估其纵向变化是否与糖尿病风险相关。本研究旨在评估两者之间的关联, 并探索肌酐与胱抑素C比值的变化与中老年人心脏代谢控制的关系。 方法: 我们共纳入了3278名来自中国健康与养老追踪调查中年龄≥45岁且提供了基线和4年后肌酐和胱抑素C测量值的参与者。糖尿病的诊断标准主要依据葡萄糖、糖化血红蛋白A1c(HbA1c)、病史或抗糖尿病药物的使用。优势比(OR)和95%置信区间(CI)使用logistic回归分析获得。 结果: 经过4年的随访, 272名参与者罹患糖尿病。肌酐与胱抑素C比值增加越多, 糖尿病风险越低。肌酐与胱抑素C比值每增加1个标准差, 经多变量校正的糖尿病OR值为 0.84(95% CI 0.72-0.98)。与肌酐与胱抑素C比值降低但体重指数(BMI)增加的参与者相比, 那些肌酐与胱抑素C比值增加和BMI降低的参与者糖尿病风险降低最大(多变量调整后的OR为0.52)。在4年随访中, 肌酐与胱抑素C比值的变化与血压、HbA1c、血脂和 C反应蛋白呈负相关。此外, 它们还与HbA1c和C反应蛋白的变化呈负相关(所有 P ≤ 0.004)。 结论: 肌酐与胱抑素C比值的增加可使得糖尿病风险降低, 并可能有益于心脏代谢控制。.

Aims and objectives To examine the effects of family functioning and resilience on self-management and glycaemic control among youth with type 1 diabetes and to determine whether resilience mediates the effects of family functioning on self-management and glycaemic control. Background Poor self-management and glycaemic control are common in youth with type 1 diabetes. Family functioning and resilience are known to be important psychosocial factors that contribute to individual health and development. However, no studies have explored the effects of family functioning and resilience on self-management and glycaemic control among youths with type 1 diabetes in mainland China. Design This study was conducted using a survey with a convenience sample following the STROBE guidelines. Methods A total of 204 Chinese youth who had been diagnosed with type 1 diabetes for at least 6 months were recruited. Family functioning, resilience, self-management and diabetes distress were measured using self-reports and standard measurement tools. Glycaemic control was assessed by glycated haemoglobin (HbA1C ) levels. A structural equation model was used to test the hypothesised model. Results The final model accounted for 52.1% and 19.5% of the total variance of self-management and HbA1C level, respectively. Resilience had a direct effect on self-management and an indirect effect on control of HbA1C . Family functioning had an indirect effect on both self-management and control of HbA1C through resilience. The model remained invariant across the mild-distress and severe-distress groups. Conclusion In Chinese youth with type 1 diabetes, resilience positively affected self-management and ultimately optimised glycaemic control, even in the presence of diabetes distress. Family functioning positively affected self-management and glycaemic control by promoting resilience. Relevance to clinical practice This study found that family functioning and resilience had positive effects on self-management and glycaemic control in youth. This study confirms the importance of incorporating resilience assessments and family-based resilience interventions into clinical nursing practice with youth with type 1 diabetes.

Formalin-fixed, paraffin-embedded (FFPE), biobanked tissue samples offer an invaluable resource for clinical and biomarker research. Here we developed a pressure cycling technology (PCT)-SWATH mass spectrometry workflow to analyze FFPE tissue proteomes and applied it to the stratification of prostate cancer (PCa) and diffuse large B-cell lymphoma (DLBCL) samples. We show that the proteome patterns of FFPE PCa tissue samples and their analogous fresh frozen (FF) counterparts have a high degree of similarity and we confirmed multiple proteins consistently regulated in PCa tissues in an independent sample cohort. We further demonstrate temporal stability of proteome patterns from FFPE samples that were stored between one to 15 years in a biobank and show a high degree of the proteome pattern similarity between two types histological region of small FFPE samples, i.e. punched tissue biopsies and thin tissue sections of micrometer thickness, despite the existence of certain degree of biological variations. Applying the method to two independent DLBCL cohorts we identified myeloperoxidase (MPO), a peroxidase enzyme, as a novel prognostic marker. In summary, this study presents a robust proteomic method to analyze bulk and biopsy FFPE tissues and reports the first systematic comparison of proteome maps generated from FFPE and FF samples. Our data demonstrate the practicality and superiority of FFPE over FF samples for proteome in biomarker discovery. Promising biomarker candidates for PCa and DLBCL have been discovered.

Background and aims Cell adhesion molecules (CAMs) are implicated in the initiation and progression of atherosclerosis, but their association with risk of type 2 diabetes remains inconsistent. This meta-analysis aimed to quantify this association with dose-response analysis in the general population without type 2 diabetes at baseline. Methods Prospective studies, investigating the association of circulating (plasma/serum) CAMs, such as intercellular adhesion molecule-1 (ICAM-1), E-selectin, vascular cell adhesion molecule-1 (VCAM-1), and P-selectin, with risk of type 2 diabetes, were included. The overall relative risks (RRs) with 95% confidence intervals (CIs) were calculated using a random-effects model. Results Sixteen datasets from 15 studies were included. The overall RR was 1.88 (95% CI 1.59 to 2.23) per 1-ln μg/ml increase in ICAM-1, and 2.44 (95% CI 1.90 to 3.12) per 1-ln μg/ml increase in E-selectin. These associations were log-linearly shaped (both pnon-linearity >0.05) and independent of traditional cardiovascular risk factors (all p Conclusions Elevated circulating CAMs, especially ICAM-1 and E-selectin, led to increased risk of type 2 diabetes in a dose-dependent manner, supporting the assumption that endothelial dysfunction contributes to the development of diabetes.

Background Evidence emerges that cardiorespiratory fitness (CRF) might be implicated in the development of heart failure (HF). This meta-analysis aimed to quantify the association between CRF exposed at baseline and HF risk with dose–response analysis and to assess whether CRF changes over time are correlated with alterations in HF risk. Methods and Results Cohort studies that assessed the association between CRF and risk of HF in subjects without baseline HF were included. Study-specific multivariate-adjusted relative risks (RRs) with 95% confidence intervals (CIs) were pooled using a random-effects model. Ten studies from 8 articles were included, enrolling 8987 incident HF cases from 154,598 participants. The RR of HF per 1-metabolic equivalent (MET) higher CRF at baseline was 0.82 (95% CI 0.80–0.84) in the overall population. The RRs were similar in men (0.82, 95% CI 0.80–0.85) and women (0.81, 95% CI 0.78–0.84), and remained minorly changed in patients with existing diabetes, hypertension, or cardiovascular disease at entry. No evidence of a nonlinear relationship between CRF at baseline and risk of HF was observed (Pnonlinearity = .18). The RR of HF per 1-MET increase in CRF over time was 0.79 (95% CI 0.67–0.93), and the measurement of CRF provided incremental value to the prediction of HF beyond conventional models. Conclusions High or increased CRF resulted in reduced risk of HF in a dose-dependent manner, supporting the necessity to increase CRF to prevent HF in clinical practice.

Background/aims The expression of transcription factor Zbtb1 is essential for the maintenance and development of various blood cells in the hematopoietic system. In the current study, we found that the total number of thymocytes in PLZF deficient mice was reduced compared with thymocytes in wild-type mice, and the number of early T-cell progenitors decreased. However, the decrease of thymocytes in PLZF deficient mice was not cell intrinsic. This study adds new information regarding the regulation of the PLZF gene in the development and self-renewal of T cells. Methods The thymus was isolated from newborn mice, and the two lobes of each thymus were physically separated. Each host received a thymus, two lobes, each placed at one end of the kidney, as described in the literature. Tail vein blood was periodically collected from some of the recipients and analyzed for the presence of peripheral blood T cells. Results In PLZF-EGFP reporter mice and neonatal thymus transplantation to the kidney, we found that PLZF was highly expressed in DN1 (Lineage-CD44+CD25-) cells of thymic grafts of Rag2/γc-/- recipient mice. We found that the proportion of PLZF wild-type and mutant-derived cells in the thymocytes of recipient mice after bone marrow transplantation is approximately equal to the competitive bone marrow chimeric mouse model, and all mice contain a normal thymus. Conclusion The development of T cells suggests that the effect of the PLZF gene on T cell differentiation and development is not cell intrinsic. However, in the neonatal mouse thymic transplant model in the Rag2/γc-/- recipient mouse, deletion of the PLZF gene results in a significant decrease in the proportion of DN1 cells from the donor in the thymic graft.

Treatment of glioblastoma (GBM) remains a challenge using conventional chemotherapy, such as temozolomide (TMZ), and is often ineffective as a result of drug resistance. We have assessed a novel nitrone-based agent, OKN-007, and found it to be effective in decreasing tumor volumes and increasing survival in orthotopic GBM xenografts by decreasing cell proliferation and angiogenesis and increasing apoptosis. In this study, we assessed combining OKN-007 with TMZ in vivo in a human G55 GBM orthotopic xenograft model and in vitro in TMZ-resistant and TMZ-sensitive human GBM cell lines. For the in vivo studies, magnetic resonance imaging was used to assess tumor growth and vascular alterations. Percent animal survival was also determined. For the in vitro studies, cell growth, IC50 values, RNA-seq, RT-PCR, and ELISA were used to assess growth inhibition, possible mechanism-of actions (MOAs) associated with combined OKN-007 + TMZ versus TMZ alone, and gene and protein expression levels, respectively. Microarray analysis of OKN-007–treated rat F98 glioma tumors was also carried out to determine possible MOAs of OKN-007 in glioma-bearing animals either treated or not treated with OKN-007. OKN-007 seems to elicit its effect on GBM tumors via inhibition of tumorigenic TGF-β1, which affects the extracellular matrix. When combined with TMZ, OKN-007 significantly increases percent survival, decreases tumor volumes, and normalizes tumor blood vasculature in vivo compared to untreated tumors and seems to affect TMZ-resistant GBM cells possibly via IDO-1, SUMO2, and PFN1 in vitro. Combined OKN-007 + TMZ may be a potentially potent treatment strategy for GBM patients.

Serum and plasma contain abundant biological information that reflect the body's physiological and pathological conditions and are therefore a valuable sample type for disease biomarkers. However, comprehensive profiling of the serological proteome is challenging due to the wide range of protein concentrations in serum. Methods: To address this challenge, we developed a novel in-depth serum proteomics platform capable of analyzing the serum proteome across ~10 orders or magnitude by combining data obtained from Data Independent Acquisition Mass Spectrometry (DIA-MS) and customizable antibody microarrays. Results: Using psoriasis as a proof-of-concept disease model, we screened 50 serum proteomes from healthy controls and psoriasis patients before and after treatment with traditional Chinese medicine (YinXieLing) on our in-depth serum proteomics platform. We identified 106 differentially-expressed proteins in psoriasis patients involved in psoriasis-relevant biological processes, such as blood coagulation, inflammation, apoptosis and angiogenesis signaling pathways. In addition, unbiased clustering and principle component analysis revealed 58 proteins discriminating healthy volunteers from psoriasis patients and 12 proteins distinguishing responders from non-responders to YinXieLing. To further demonstrate the clinical utility of our platform, we performed correlation analyses between serum proteomes and psoriasis activity and found a positive association between the psoriasis area and severity index (PASI) score with three serum proteins (PI3, CCL22, IL-12B). Conclusion: Taken together, these results demonstrate the clinical utility of our in-depth serum proteomics platform to identify specific diagnostic and predictive biomarkers of psoriasis and other immune-mediated diseases.

The precise control of neural activities at both cellular and circuit levels reveals significant impacts on the fundamental neuroscience explorations and medical applications. Optogenetic methods provide efficient cell-specific modulations, and the ability of simultaneous neural activation and inhibition in the same brain region of freely moving animals is highly desirable and being actively researched. Here we report bidirectional neuronal activity manipulation accomplished by a wireless, dual-color optogenetic probe in synergy with the co-expression of two spectrally distinct opsins (ChrimsonR and stGtACR2) in a rodent model. Based on vertically assembled, thin-film microscale light-emitting diodes (micro-LEDs) with a lateral dimension of 125 × 180 µm2 on flexible substrates, the dual-color probe shows colocalized red and blue emissions and allows chronic in vivo operations with desirable biocompatibilities. In addition, we discover that neurons co-expressing the two opsins can be deterministically evoked or silenced under red or blue irradiations. Implanted in behaving mice, the wirelessly controlled dual-color probe interferes with dopaminergic neurons in the ventral tegmental area (VTA), increasing or decreasing dopamine levels with colocalized red and blue stimulations. Such bidirectional regulations further generate rewarding and aversive behaviors of freely moving mice in a place preference test and interrogate social interactions among multiple mice. The technologies established here will create numerous opportunities and profound implications for brain research.

The biosynthetic pathway of l -methionine in microorganisms was complex and regulated at multiple levels. In this study, a two-step method for l -methionine production combined fermentation and biocatalysis was realized in one pot. The O-succinyl- l -homoserine (OSH) producing strain Escherichia coli W3110(DE3) ΔIJB∗TrcmetL/pTrc-metAfbr-Trc-thrAfbr-yjeH (ΔIJB) was constructed initially. OSH in the fermentation supernatant was then converted to l -methionine in the presence of O-succinyl- l -homoserine sulfhydrylase (OSHS) and sodium methanethiol. The titer of l -methionine could reach 21.1 g/L after 88 h (84 h fermentation and 4 h catalysis) in a two-step method (process 1). In a one-pot two-strain system (process 2), two strains ΔIJB and E. coli BL21(DE3)/pET28b–OSHS–cutinase were co-cultured, and 8.24 g/L l -methionine was obtained. In another one-pot one-strain system (process 3), strain E. coli ΔIJB/pET28b–OSHS–cutinase could co-express OSHS and cutinase during ΔIJB fermentation at the same time, obtaining 13.6 g/L l -methionine in a 5 L fermentor after 84 h. By comparing the three processes for l -methionine production based on the process 1, the simplified process in process 3 provided in this study showed potent in the large-scale production of l -methionine with convenient handling and production efficiency, but further works still need to be carried out to improve the l -methionine production.

Background and aims Estimated cardiorespiratory fitness (eCRF) derived from algorithm correlates well with exercise testing-measured CRF, yet its clinical use for mortality risk stratification has not been systematically evaluated. This meta-analysis with dose-response analysis was conducted to quantify its association with risk of cardiovascular and all-cause mortality. Methods Electronic databases were searched for prospective cohort studies that investigated the association of eCRF with risk of cardiovascular and all-cause mortality. Study-specific multivariate-adjusted hazard ratios (HRs) and 95% confidence intervals (CIs) per 1-metabolic equivalent (MET) higher of eCRF were pooled using a random-effects model. Results Twenty-five datasets from 8 cohort studies that enrolled more than 170,000 participants were included. The summary HR per 1-MET higher of eCRF was 0.83 (95% CI 0.80 to 0.86) for cardiovascular mortality (11 datasets) and 0.83 (95% CI 0.78 to 0.88) for all-cause mortality (14 datasets) in the general population. These associations showed no sex-difference and were all linearly shaped (all pnonlinearity ≥ 0.27). The performance of eCRF (assessed by the area under the curve) in discriminating future risk of cardiovascular and all-cause mortality was higher than all its components (such as physical activity, resting heart rate, and body mass index, all p Conclusions Higher eCRF was independently associated with lower risk of cardiovascular and all-cause mortality in the general population, indicating that eCRF might hold the potential as an effective and practical risk prediction tool in epidemiological or population research.

This study aimed to compare the gene expression variation of clinical primary osteosarcoma (OS) and metastatic OS, identify expression profiles and signal pathways related to disease classification, and systematically evaluate the potential anticancer effect and molecular mechanism of ginsenoside Rh2 on OS. A raw dataset (GSE14359), which excluded GSM359137 and GSM359138, was downloaded from the Gene Expression Omnibus. Differentially expressed genes (DEGs) and principal component analysis (PCA) were obtained with limma. Pathways enrichment analysis was understood by GSEA app. Rh2-associated targets were harvested and mapped through PharmMapper and Cytoscape 3.4.0. The toxicity of Rh2 was determined using crystal staining and MTT assay on 143B and MG63 cell lines. The relative protein expression was confirmed through Western blot analysis. The mitochondrial membrane potential (△Ψm) was evaluated by JC-1 fluorescence staining. The cell mobility was measured via wound healing and transwell assays. A total of 752 genes were upregulated, while 161 genes were downregulated. GSEA and PCA displayed significant function enrichment and classification. Through PharmMapper and Cytoscape 3.4.0, Rh2 was found to target the mitogen activated protein kinase (MAPK) and PI3K signaling pathways, which are the key pathways in the metastasis of OS. Furthermore, Rh2 induced a concentration-dependent decrease in cell viability and early apoptosis associated with ΔΨm decline, while a non-lethal dose of Rh2 weakened the metastatic capability. Moreover, systematic evaluation showed that promoting the MAPK signaling pathway and inhibiting PI3K/Akt/mTOR were correlated with the anticancer effects of Rh2 on metastatic OS. In conclusion, transcriptome-derived approaches may be beneficial in diagnosing early metastases, and Rh2, a multi-targeting agent, shows promising application potential in suppressing metastatic OS in an MAPK- and PI3K/Akt/mTOR-dependent manner.

The diagnosis and disease course monitoring of COVID-19 are mainly based on RT-PCR analysis of RNAs extracted from pharyngeal or nasopharyngeal swabs with potential live virus, posing a high risk to medical practitioners. Here, we investigated the feasibility of applying serum proteomics to classify COVID-19 patients in the nucleic acid positive (NCP) and negative (NCN) stages. We analyzed the proteome of 320 inactivated serum samples from 144 COVID-19 patients, and 45 controls and shortlisted 42 regulated proteins in the severe group and 12 regulated proteins in the non-severe group. Together with several key clinical indexes including days after symptom onset, platelet counts and magnesium, we developed machine learning models to classify NCP and NCN with an AUC of 0.94 for the severe cases and 0.89 for the non-severe cases. This study suggests the feasibility of utilizing quantitative serum proteomics for NCP-NCN classification. Funding: This work was supported by grants from the National Key R&D Program of China(No. 2020YFE0202200), National Natural Science Foundation of China (81672086), Zhejiang Province Analysis Test Project (2018C37032), the National Natural Science Foundation of China (81972492, 21904107), Zhejiang Provincial Natural Science Foundation for Distinguished Young Scholars (LR19C050001), Zhejiang Medical and Health Science and Technology Plan (2021KY394), Hangzhou Agriculture andSociety Advancement Program (20190101A04), and Westlake Education Foundation, Tencent Foundation. Conflict of Interest: Tiannan Guo is shareholder of Westlake Omics Inc. W.G. and N.X. are employees of Westlake Omics Inc. The remaining authors declare no competing interests. Ethical Approval: This study has been approved by both the Ethical/Institutional Review Boards of Taizhou Hospital and Westlake University. Informed contents from patients were waived by the boards.

Background/Aims: Older adults are vulnerable to central obesity, while the association of changes in central fatness with risk of diabetes and metabolic control has not been investigated among this particular population. This study was aimed to address these issues.Methods: A total of 1,815 adults aged ≥ 60 years without diabetes at baseline were followed for 4 years. Incident diabetes was ascertained based on plasma glucose, hemoglobin A1c, medical history, and/or the use of anti-diabetic drugs. Central fatness was assessed by waist circumference (WC), waist-height ratio (WHtR), and body roundness index (BRI). Logistic regression analyses were used to assess the association of changes in central fatness with risk of diabetes, along with dose-response and mediation analyses.Results: During the 4-year follow-up, 177 participants developed diabetes. The risk of diabetes was increased by 42%, 41%, and 40% per 1 standard deviation increases in WC, WHtR, and BRI, respectively, in multivariable-adjusted models (all p < 0.01). Moreover, these relationships were all linearly-shaped (all pnonlinearity ≥ 0.11). Increases in WC, WHtR, and BRI correlated with increases in hemoglobin A1c, triglycerides-and-glucose index, triglycerides, white blood cell, and C-reactive protein (all p ≤ 0.04). Yet only changes in hemoglobin A1c and triglycerides-and-glucose index were identified as the possible mediators for risk of diabetes, with their mediating effect being about 35% and 21%, respectively.Conclusions: Increases in central fatness were related to elevated risk of diabetes, and this association might be partly explained by the worsening of glycemic control and insulin resistance in older adults.

Aim: Diabetic peripheral neuropathy (DPN) is a common, severe microvascular complication of diabetes. Our study was to assess prevalence and risk factors for DPN in subjects with type 2 diabetes from 14 different countries. Methods: A total of 2,733 subjects with type 2 diabetes aged 18–65 years (45.3% men, mean duration of diabetes = 8.8 years) were included to perform this International Prevalence and Treatment of Diabetes and Depression (INTERPRET-DD) study in 14 countries. After a structured questionnaire was used in face-to-face interviews to collect sociodemographic characteristics and medical records of the participating subjects, laboratory tests were carried out for clinical measurement. Depressive symptoms were diagnosed and measured using the Patient Health Questionnaire-9. The potential risk factors for DPN were determined by multilevel mixed-effects logistic regression, accounting for clustering of participants within the country. Robustness of the estimates was assessed by sensitivity analysis. Results: The overall prevalence of DPN across different countries was 26.71%, whereas country-specific prevalences showed considerable variation. Multivariate analysis revealed that duration of diabetes (OR: 1.08 per 1-year increase, 95% CI: 1.06–1.09), poor glycemic control (OR: 1.11 per 1% increase in HbA1c, 95% CI: 1.05–1.18), and history of hypertension (OR: 1.58, 95% CI: 1.18–2.12), cardiovascular disease (OR: 2.07, 95% CI: 1.55–2.78) and depressive symptoms (OR: 1.92, 95% CI: 1.43–2.58) were independently and positively associated with the risk of DPN. Sensitivity analyses including or excluding patients from countries with extreme low or high prevalence of DPN yielded similar estimates in terms of trend and magnitude. Conclusions: This international study illustrates that more than a quarter of individuals with type 2 diabetes developed DPN. The prevalence was positively associated with the duration of diabetes, poor glycemic control, and history of hypertension, cardiovascular disease and depressive symptoms.

Glioblastoma (GBM) is one of the most malignant primary brain tumors. This neoplasm is the hardest to treat and has a bad prognosis. Because of the characteristics of genetic heterogeneity and frequent recurrence, a successful cure for the disease is unlikely. Increasing evidence has revealed that the GBM stem cell-like cells (GSCs) and microenvironment are key elements in GBM recurrence and treatment failure. To better understand the mechanisms underlying this disease and to develop more effective therapeutic strategies for treatment, suitable approaches, techniques, and model systems closely mimicking real GBM conditions are required. Microfluidic devices, a model system mimicking the in vivo brain microenvironment, provide a very useful tool to analyze GBM cell behavior, their correlation with tumor malignancy, and the efficacy of multiple drug treatment. This paper reviews the applications of microfluidic devices in GBM research and summarizes progress and perspectives in this field.

Background: The COVID-19 pandemic is spreading globally with high disparity in the susceptibility of the disease severity. Identification of the key underlying factors for this disparity is highly warranted. Results: Here we describe constructing a proteomic risk score (PRS) based on 20 blood proteomic biomarkers which related to the progression to severe COVID-19. Among COVID-19 patients, per 10% increment in the PRS was associated with a 57% higher risk of progressing to clinically severe phase (RR=1.57; 95% CI, 1.35-1.82). We demonstrate that in our own cohort of 990 individuals without infection, this proteomic risk score is positively associated with proinflammatory cytokines mainly among older, but not younger, individuals. We further discovered that a core set of gut microbiota could accurately predict the blood proteomic biomarkers of COVID-19 using a machine learning model. The core OTU-predicted PRS had a significant correlation with actual PRS both cross-sectionally (n=132, pConclusions: Our study suggests that gut microbiota may underlie the predisposition of healthy individuals to COVID-19-sensitive proteomic biomarkers.

Bradyarrhythmias are an important cause of mortality in heart failure and previous studies indicate a mechanistic role for electrical remodelling of the key pacemaking ion channel HCN4 in this process. Here we show that, in a mouse model of heart failure in which there is sinus bradycardia, there is upregulation of a microRNA (miR-370-3p), downregulation of the pacemaker ion channel, HCN4, and downregulation of the corresponding ionic current, If, in the sinus node. In vitro, exogenous miR-370-3p inhibits HCN4 mRNA and causes downregulation of HCN4 protein, downregulation of If, and bradycardia in the isolated sinus node. In vivo, intraperitoneal injection of an antimiR to miR-370-3p into heart failure mice silences miR-370-3p and restores HCN4 mRNA and protein and If in the sinus node and blunts the sinus bradycardia. In addition, it partially restores ventricular function and reduces mortality. This represents a novel approach to heart failure treatment.

Summary Early detection and effective treatment of severe COVID-19 patients remain major challenges. Here, we performed proteomic and metabolomic profiling of sera from 46 COVID-19 and 53 control individuals. We then trained a machine learning model using proteomic and metabolomic measurements from a training cohort of 18 non-severe and 13 severe patients. The model was validated using 10 independent patients, 7 of which were correctly classified. Targeted proteomics and metabolomics assays were employed to further validate this molecular classifier in a second test cohort of 19 COVID-19 patients, leading to 16 correct assignments. We identified molecular changes in the sera of COVID-19 patients compared to other groups implicating dysregulation of macrophage, platelet degranulation, complement system pathways, and massive metabolic suppression. This study revealed characteristic protein and metabolite changes in the sera of severe COVID-19 patients, which might be used in selection of potential blood biomarkers for severity evaluation., Graphical Abstract, Highlights • 93 proteins show differential expression in severe COVID-19 patient sera • 204 metabolites in COVID-19 patient sera correlate with disease severity • A model composed of 29 serum factors shows patient stratification potential • Pathway analysis highlights metabolic and immune dysregulation in COVID-19 patients, Proteomic and metabolomic analysis of COVID-19 sera identifies differentially expressed factors that correlate with disease severity and highlights dysregulation of multiple immune and metabolic components in clinically severe patients.

Little is known regarding why a subset of COVID-19 patients exhibited prolonged positivity of SARS-CoV-2 infection. Here, we studied the sera proteomic dynamics in 37 COVID-19 patients over nine weeks, quantifying 2700 proteins with high quality. Remarkably, we found that during the first three weeks since disease onset, while clinical symptoms and outcome were indistinguishable, patients with prolonged disease course displayed characteristic immunological responses including enhanced Natural Killer cell-mediated innate immunity and regulatory T cell-mediated immunosuppression. We further showed that it is possible to predict the length of disease course using machine learning based on blood protein levels during the first three weeks. Validation in an independent cohort achieved an accuracy of 82%. In summary, this study presents a rich serum proteomic resource to understand host responses in COVID-19 patients and identifies characteristic Treg-mediated immunosuppression in patients with prolonged disease course, nominating new therapeutic target and diagnosis strategy.

SUMMARYThe COVID-19 pandemic is spreading globally with high disparity in the susceptibility of the disease severity. Identification of the key underlying factors for this disparity is highly warranted. Here we describe constructing a proteomic risk score based on 20 blood proteomic biomarkers which predict the progression to severe COVID-19. We demonstrate that in our own cohort of 990 individuals without infection, this proteomic risk score is positively associated with proinflammatory cytokines mainly among older, but not younger, individuals. We further discovered that a core set of gut microbiota could accurately predict the above proteomic biomarkers among 301 individuals using a machine learning model, and that these gut microbiota features are highly correlated with proinflammatory cytokines in another set of 366 individuals. Fecal metabolomic analysis suggested potential amino acid-related pathways linking gut microbiota to inflammation. This study suggests that gut microbiota may underlie the predisposition of normal individuals to severe COVID-19.

SUMMARYUp to 30% of thyroid nodules cannot be accurately classified as benign or malignant by cytopathology. Diagnostic accuracy can be improved by nucleic acid-based testing, yet a sizeable number of diagnostic thyroidectomies remains unavoidable. In order to develop a protein classifier for thyroid nodules, we analyzed the quantitative proteomes of 1,725 retrospective thyroid tissue samples from 578 patients using pressure-cycling technology and data-independent acquisition mass spectrometry. With artificial neural networks, a classifier of 14 proteins achieved over 93% accuracy in classifying malignant thyroid nodules. This classifier was validated in retrospective samples of 271 patients (91% accuracy), and prospective samples of 62 patients (88% accuracy) from four independent centers. These rapidly acquired proteotypes and artificial neural networks supported the establishment of an effective protein classifier for classifying thyroid nodules.

To answer the increasing need for detecting and validating protein biomarkers in clinical specimens, proteomic techniques are required that support the fast, reproducible and quantitative analysis of large clinical sample cohorts. Targeted mass spectrometry techniques, specifically SRM, PRM and the massively parallel SWATH/DIA technique have emerged as a powerful method for biomarker research. For optimal performance, they require prior knowledge about the fragment ion spectra of targeted peptides. In this report, we describe a mass spectrometric (MS) pipeline and spectral resource to support data-independent acquisition (DIA) and parallel reaction monitoring (PRM) based biomarker studies. To build the spectral resource we integrated common open-source MS computational tools to assemble an open source computational workflow based on Docker. It was then applied to generate a comprehensive DIA pan-human library (DPHL) from 1,096 data dependent acquisition (DDA) MS raw files, and it comprises 242,476 unique peptide sequences from 14,782 protein groups and 10,943 SwissProt-annotated proteins expressed in 16 types of cancer samples. In particular, tissue specimens from patients with prostate cancer, cervical cancer, colorectal cancer, hepatocellular carcinoma, gastric cancer, lung adenocarcinoma, squamous cell lung carcinoma, diseased thyroid, glioblastoma multiforme, sarcoma and diffuse large B-cell lymphoma (DLBCL), as well as plasma samples from a range of hematologic malignancies were collected from multiple clinics in China, the Netherlands and Singapore and included in the resource. This extensive spectral resource was then applied to a prostate cancer cohort of 17 patients, consisting of 8 patients with prostate cancer (PCa) and 9 with benign prostate hyperplasia (BPH), respectively. Data analysis of DIA data from these samples identified differential expressions of FASN, TPP1 and SPON2 in prostate tumors. Thereafter, PRM validation was applied to a larger PCa cohort of 57 patients and the differential expressions of FASN, TPP1 and SPON2 in prostate tumors were validated. As a second application, the DPHL spectral resource was applied to a patient cohort consisting of samples from 19 DLBCL patients and 18 healthy individuals. Differential expressions of CRP, CD44 and SAA1 between DLBCL cases and healthy controls were detected by DIA-MS and confirmed by PRM. These data demonstrate that the DPHL supported that DIA-PRM MS pipeline enables robust protein biomarker discoveries.

Background: Early screening for colorectal cancer (CRC) is essential to improve its prognosis. Liquid biopsies are increasingly being considered for diagnosing cancer due to low invasiveness and high reproducibility. In addition, circulating extracellular vesicles (crEVs, extracellular vesicles isolated from plasma) expressing tumour-specific proteins are potential biomarkers for various cancers. Here, we present a data-independent acquisition (DIA)-mass spectrometry (MS)-based diagnostic method for liquid biopsies. Methods: Extracellular vesicles (EVs) were isolated from culture supernatants of human CRC cell lines, and plasma of patients with CRC at different tumour stages, by overnight ultracentrifugation coupled with sucrose density gradient centrifugation. Tumour-specific EV proteins were prioritized using Tandem Mass Tag (TMT)-based shotgun proteomics and phosphoproteomics. The results were verified in a second independent cohort and a mouse tumour-bearing model using Western blotting (WB). The candidate biomarkers were further validated in a third cohort by DIA-MS. Finally, the DIA-MS methodology was accelerated to permit high-throughput detection of EV biomarkers in another independent cohort of patients with CRC and healthy controls. Results: High levels of total and phosphorylated fibronectin 1 (FN1) in crEVs, haptoglobin (HP), S100A9 and fibrinogen α chain (FGA) were significantly associated with cancer progression. FGA was the most dominant biomarker candidate. Analysis of the human CRC cell lines and the mouse model indicated that FGA+ crEVs were likely released by CRC cells. Furthermore, fast DIA-MS and parallel reaction monitoring (PRM)-MS both confirmed that FGA+ crEVs could distinguish colon adenoma with an area of curve (AUC) in the receiver operating characteristic (ROC) curve of 0.949 and patients with CRC (AUC of ROC is 1.000) from healthy individuals. The performance outperformed conventional tumour biomarkers. The DIA-MS quantification of FGA+ crEVs among three groups agreed with that from PRM-MS. Conclusion: DIA-MS detection of FGA+ crEVs is a potential rapid and non-invasive screening tool to identify early stage CRC. Abbreviations: FGA: fibrinogen α chain; CRC: colorectal cancer; crEVs: circulating extracellular vesicles; EV: extracellular vesicles;MS: mass spectrometry; WB: Western blotting; ROC: receiver operating characteristic; PRM: Parallel Reaction Monitoring; GPC1: Glypican-1; GO: Gene ontology; TEM: transmission electron microscopy; FN1: Fibronectin 1; HP: haptoglobin; TMT: Tandem Mass Tag; LC-MS/MS: liquid chromatography coupled to tandem mass spectrometry; DIA: data-independent acquisition; DDA: data-dependent acquisition; CiRT: Common internal Retention Time standards;AGC: Automatic gain control; AUC: area under curve.

Little is known regarding why a subset of COVID-19 patients exhibited prolonged positivity of SARS-CoV-2 infection. Here, we present a longitudinal sera proteomic resource for 37 COVID-19 patients over nine weeks, in which 2700 proteins were quantified with high quality. Remarkably, we found that during the first three weeks since disease onset, while clinical symptoms and outcome were indistinguishable, patients with prolonged disease course displayed characteristic immunological responses including enhanced Natural Killer (NK) cell-mediated innate immunity and regulatory T cell-mediated immunosuppression. We further showed that it is possible to predict the length of disease course using machine learning based on blood protein levels during the first three weeks. Validation in an independent cohort achieved an accuracy of 82%. In summary, this study presents a rich serum proteomic resource to understand host responses in COVID-19 patients and identifies characteristic Treg-mediated immunosuppression in LC patients, nominating new therapeutic target and diagnosis strategy.

Macrophages, which are the main regulators of the tumor-associated microenvironment, play a crucial role in the progression of various tumors. The anti-inflammatory role of β-catenin in macrophages has been extensively studied in recent years. However, the association between macrophages and β-catenin with regards to the development of glioma has not yet been investigated, at least to the best of our knowledge. The present study found that fibroblast growth factor 20 (FGF20), as a paracrine cytokine, was secreted by glioma cells and acted on macrophages. FGF20 treated macrophages exhibited a decreased pro-inflammatory phenotype upon LPS and IFN-γ stimulation, characterized by the decreased the level of M1 macrophage markers and the reduced production of pro-inflammatory cytokines. Mechanistic analysis revealed that FGF20 interacted with FGF receptor 1 isoform of macrophages, and subsequently increased the stability of β-catenin via phosphorylating GSK3β, which suppressed macrophage polarization to the M1-phenotype. Finally, it was found that FGF20 of glioma cells expression was upregulated by the glucocorticoids (GCs) treatment, and decreased FGF20 expression of glioma cells markedly blocked the effects of GCs on the polarization of macrophages. On the whole, the present study demonstrates that FGF20, secreted from glioma cells, participates the GCs regulated macrophage function and exerts anti-inflammatory effects during the treatment of glioma by GCs. Moreover, a molecular link was identified between glioma cells and macrophages, demonstrating that FGF20 modulates the GCs-induced dysfunction of macrophages during glioma development.

The present study examined the effects of irrigant flow rate and simulated intraosseous sinusoidal pressure on the rate of NaOCl extrusion from the apical terminus of a faux root canal.An extrusion setup was designed to enable irrigant extrusion to be opposed by 30 mm Hg simulated intraosseous pressure. The faux canal apex was opposed by atmospheric + 30 mm Hg pressure (experimental) or atmospheric pressure only (control group). Using five irrigant delivery rates (15.6 8.0, 4.0, 3.4 or 3.0 mL/min), the extrusion rates of 2% NaOCl from the faux apex were measured in both groups (n = 16). Data were analysed with two-factor ANOVA and pairwise comparisons at α = 0.05. Correlation between NaOCl delivery rates and extrusion rates in both groups were analysed with the Pearson product-moment procedure.Irrespective of the presence or absence of simulated sinusoidal pressure, NaOCl extrusion rates were positively-correlated with irrigant flow rates. For the factor "irrigant flow rates", significant differences in NaOCl extrusion rates were identified among all flow rates (p 0.05), except for the pairwise comparison between 4.0 and 3.4 mL/min in the control. For all irrigant flow rates, NaOCl extrusion rate was significantly lower in the presence of 30 mm Hg simulated sinusoidal pressure than that obtained in the absence of opposing pressure (p 0.05).In the presence of 30 mm Hg simulated intraosseous pressure, NaOCl delivered via a side-vented needle inserted to 1 mm short of working length may be prevented from extrusion when its flow rate is ≤ 3.0 mL/min.When opposed by intraosseous sinusoidal pressure, NaOCl delivered via a side-vented needle inserted to 1 mm short of working length may be prevented from extrusion when its flow rate is ≤ 3.0 mL/min.

There is growing interest in exploring circulating (plasma/serum) irisin in polycystic ovary syndrome (PCOS) patients. This meta-analysis aimed to summarize the evidence assessing circulating irisin changes in this population. A systematic search was conducted in three databases: PubMed, Cochrane Library and Web of Science, for studies reporting irisin in PCOS patients compared with healthy controls or stratified by body mass index (BMI), or assessing irisin response to hyperinsulinemia. Effect sizes (Cohen's d with 95% confidence intervals [CI]) were calculated using random-effects models. Eight studies with 918 PCOS patients and 528 healthy controls were included. Results showed that circulating irisin was higher in PCOS patients than in overall healthy controls ( d = 0.37, 95% CI 0.05 to 0.70), but not compared with BMI-matched or age- and BMI-matched controls. Circulating irisin was higher in PCOS patients with higher BMI than lower ( d = 0.36, 95% CI 0.16 to 0.56). Circulating irisin decreased 2 h later in response to euglycemic hyperinsulinemia in PCOS patients with a larger magnitude than healthy controls ( d = −0.32, 95% CI −0.53 to −0.11). In summary, with adjustment for BMI, circulating irisin in PCOS patients seems comparable to healthy controls, but its response to hyperinsulinemia might be impaired.

Background D-Pantothenate (DPA) is an important functional chemical that has been widely applied in healthcare, cosmetics, animal food and feed industries. Methods and results In this study, a high-yield DPA-producing strain was constructed by metabolic engineering strategies with targeting metabolic driving and by-products minimization. The metabolic driving force of push and pull was firstly obtained to improve the production of DPA via enrichment of precursor pool and synthetic pathway, accumulating 4.29 g L-1 DPA in shake flask fermentation. To eliminate the metabolic pressure on DPA production, an amino throttling system was proposed and successfully attenuated the synthesis of four competitive amino acids by a single-step regulation of gdhA. Further minimization of acetate was carried out by pta deletion, and utilization of β-alanine was improved via enhancing its uptake system with producing 5.78 g L-1 DPA. Finally, the engineered strain produced 66.39 g L-1 DPA with β-alanine addition in fermentor under fed-batch fermentation. Conclusion This study paved a foundation for the industrial production of DPA. This article is protected by copyright. All rights reserved.

Glioblastoma (GBM) is the most invasive and devastating primary brain tumor with a median overall survival rate about 18 months with aggressive multimodality therapy. Its unique characteristics of heterogeneity, invasion, clonal populations maintaining stem cell-like cells and recurrence, have limited responses to a variety of therapeutic approaches, and have made GBM the most difficult brain cancer to treat. A great effort and progress has been made to reveal promising molecular mechanisms to target therapeutically. Especially with the emerging of new technologies, the mechanisms underlying the pathology of GBM are becoming more clear. The purpose of this review is to summarize the current knowledge of molecular mechanisms of GBM and highlight the novel strategies and concepts for the treatment of GBM.

Background: Increased physical activity (PA) is a key element in the management of patients with nonalcoholic fatty liver disease (NAFLD); however, its association with NAFLD risk has not been systematically assessed. This meta-analysis of observational studies was to quantify this association with dose–response analysis. Methods: Electronic databases were searched to January 2017 for studies of adults reporting the risk of NAFLD in relation to PA with cohort or case-control designs. Studies that reported sex-specific data were included as separate studies. The overall risk estimates were pooled using a random-effects model, and the dose–response analysis was conducted to shape the quantitative relationship. Results: A total of 6 cohort studies from 5 articles with 32,657 incident NAFLD cases from 142,781 participants, and 4 case-control studies from 3 articles with 382 NAFLD cases and 302 controls were included. Compared with the lowest PA level, the highest PA level was associated with a risk reduction of NAFLD in cohort [RR (risk ratio) 0.79, 95% CI (confidence interval) 0.71–0.89] and case-control studies [OR (odds ratio) 0.43, 95% CI 0.27–0.68]. For cohort studies, both highest and moderate PA levels were superior to the light one in lowering NAFLD risk ( pfor interaction = 0.006 and 0.02, respectively), and there was a log-linear dose–response association ( pfor nonlinearity = 0.10) between PA and NAFLD risk [RR 0.82 (95% CI 0.73–0.91) for every 500 metabolic equivalent (MET)-minutes/week increment in PA]. Conclusions: Increased PA may lead to a reduced risk of NAFLD in a dose-dependent manner, and the current guideline-recommended minimum PA level that approximates to 500 MET-minutes/week is able to moderately reduce the NAFLD risk.

Stroke is a leading cause of mortality and disability worldwide. There is growing evidence that metformin (Met) has potent neuroprotective effects; however, its mechanisms remain unclear. We examined the role of the arterial baroreflex and cholinergic-α7 nicotinic acetylcholine receptor (α7nAChR) anti-inflammory pathway in the beneficial effects of Met against stroke. Stroke-prone spontaneously hypertensive rats (SHRSP) were used to observe stroke development indicated by lifespan of SHRSP and the ischemic injury induced by permanent middle cerebral artery occlusion (MCAO). Sinoaortic denervation was used to inactivate the arterial baroreflex. MCAO were also performed in α7nAChR knockout (KO) mice. Briefly, Met increased the life span of SHRSP and reduced the infarct area induced by MCAO. Met also improved the function of arterial baroreflex. The beneficial effects of Met on stroke were markedly attenuated by blunting the arterial baroreflex. Met up-regulated the expression of vesicular acetylcholine transporter (VAChT) and α7nAChR, down-regulated the level of pro-inflammtory cytokines in serum and peri-infarct of ischemic brain. Arterial baroreflex dysfunction decreased the expression of VAchT and α7nAChR, showed upward tendency in the level of pro-inflammtory cytokines. Most importantly, arterial baroreflex dysfunction nearly abolished such effect of Met on cholinergic signaling. In addition, the α7nAChR KO mice also had significantly worse ischemic damage induced by MCAO, and neuroprotection of Met disappeared in α7nAChR KO mice. In conclusion, Met improved the arterial baroreflex function, and then enhancing cholinergic anti-inflammatory pathway in an α7nAChR-dependent manner, thereby effectively prevent ischemic induced brain injury and delayed stroke onset in SHRSP.

(R)-Mandelic acid (R-MA) is a key precursor for the synthesis of semi-synthetic penicillin, cephalosporin, anti-obesity drugs, antitumor agents, and chiral resolving agents for the resolution of racemic alcohols and amines. In this study, an enzymatic method for the large-scale production of R-MA by a stereospecific nitrilase in an aqueous system was developed. The nitrilase activity of the Escherichia coli BL21(DE3)/pET-Nit whole cells reached 138.6 U/g in a 20,000-L fermentor. Using recombinant E. coli cells as catalyst, 500 mM R,S-mandelonitrile (R,S-MN) was resolved into 426 mM (64.85 g/L) R-MA within 8 h, and the enantiomeric excess (ee) value of R-MA reached 99%. During the purification process, pure R-MA with a recovery rate of 78.8% was obtained after concentration and crystallization. This study paved the foundation for the upscale production of R-MA using E. coli whole cells as biocatalyst.