Building RNA-Mediated Artificial Signaling Pathways between Endogenous Genes.

ConspectusSophisticated genetic networks play a pivotal role in orchestrating cellular responses through intricate signaling pathways across diverse environmental conditions. Beyond the inherent complexity of natural cellular signaling networks, the construction of artificial signaling pathways (ASPs) introduces a vast array of possibilities for reshaping cellular responses, enabling programmable control of living organisms. ASPs can be integrated with existing cellular networks and redirect output responses as desired, allowing seamless communication and coordination with other cellular processes, thereby achieving designable transduction within cells. Among diversified ASPs, establishing connections between originally independent endogenous genes is of particular significance in modifying the genetic networks, so that cells can be endowed with new capabilities to sense and deal with abnormal factors related to differentiated gene expression (i.e., solve the issues of the aberrant gene expression induced by either external or internal stimuli). In a typical scenario, the two genes X and Y in the cell are originally expressed independently. After the introduction of an ASP, changes in the expression of gene X may exert a designed impact on gene Y, subsequently inducing the cellular response related to gene Y. If X represents a disease signal and Y serves as a therapeutic module, the introduction of the ASP empowers cells with a new spontaneous defense system to handle potential risks, which holds great potential for both fundamental and translational studies.In this Account, we primarily review our endeavors in the construction of RNA-mediated ASPs between endogenous genes that can respond to differentiated RNA expression. In contrast to other molecules that may be restricted to specific pathways, synthetic RNA circuits can be easily utilized and expanded as a general platform for constructing ASPs with a high degree of programmability and tunability for diversified functionalities through predictable Watson-Crick base pairing. We first provide an overview of recent advancements in RNA-based genetic circuits, encompassing but not limited to utilization of RNA toehold switches, siRNA and CRISPR systems. Despite notable progress, most reported RNA circuits have to contain at least one exogenous RNA X as input or one engineered RNA Y as a target, which is not suitable for establishing endogenous gene connections. While exogenous RNAs can be engineered and controlled as desired, constructing a general and efficient platform for manipulation of naturally occurring RNAs poses a formidable challenge, especially for the mammalian system. With a focus on this goal, we are devoted to developing efficient strategies to manipulate cell responses by establishing RNA-mediated ASPs between endogenous genes, particularly in mammalian cells. Our step-by-step progress in engineering customized cell signaling circuits, from bacterial cells to mammalian cells, from gene expression regulation to phenotype control, and from small RNA to long mRNA of low abundance and more complex secondary structures, is systematically described. Finally, future perspectives and potential applications of these RNA-mediated ASPs between endogenous genes are also discussed.

Establishing artificial gene connections through RNA displacement-assembly-controlled CRISPR/Cas9 function.

Construction of synthetic circuits that can reprogram genetic networks and signal pathways is a long-term goal for manipulation of biosystems. However, it is still highly challenging to build artificial genetic communications among endogenous RNA species due to their sequence independence and structural diversities. Here we report an RNA-based synthetic circuit that can establish regulatory linkages between expression of endogenous genes in both Escherichiacoli and mammalian cells. This design employs a displacement-assembly approach to modulate the activity of guide RNA for function control of CRISPR/Cas9. Our experiments demonstrate the great effectiveness of this RNA circuit for building artificial connections between expression of originally unrelated genes. Both exogenous and naturally occurring RNAs, including small/microRNAs and long mRNAs, are capable of controlling expression of another endogenous gene through this approach. Moreover, an artificial signal pathway inside mammalian cells is also successfully established to control cell apoptosis through our designed synthetic circuit. This study provides a general strategy for constructing synthetic RNA circuits, which can introduce artificial connections into the genetic networks of mammalian cells and alter the cellular phenotypes.

Chemically Cross-Linked Hammerhead Ribozyme as an Efficient RNA Interference Tool.

RNA-cleaving ribozymes are promising candidates as general tools of RNA interference (RNAi) in gene manipulation. However, compared with other RNA systems, such as siRNA and CRISPR technologies, the ribozyme tools are still far from broad applications on RNAi due to their poor performance in the cellular context. In this work, we report an efficient RNAi tool based on chemically modified hammerhead ribozyme (HHR). By the introduction of an intramolecular linkage into the minimal HHR to reconstruct the distal interaction within the tertiary ribozyme structure, this cross-linked HHR exhibits efficient RNA substrate cleavage activities with almost no sequence constraint. Cellular experiments suggest that both exogenous and endogenous RNA expression can be dramatically knocked down by this HHR tool with levels comparable to those of siRNA. Unlike the widely applied protein-recruiting RNA systems (siRNA and CRISPR), this ribozyme tool functions solely on RNA itself with great simplicity, which may provide a new approach for gene manipulation in both fundamental and translational studies.

Design and rationale of the Comprehensive intelligent Hypertension managEment SyStem (CHESS) evaluation study: A cluster randomized controlled trial for hypertension management in primary care.

BACKGROUND: Hypertension management in China is suboptimal with high prevalence and low control rate due to various barriers, including lack of self-management awareness of patients and inadequate capacity of physicians. Digital therapeutic interventions including mobile health and computational device algorithms such as clinical decision support systems (CDSS) are scalable with the potential to improve blood pressure (BP) management and strengthen the healthcare system in resource-constrained areas, yet their effectiveness remains to be tested. The aim of this report is to describe the protocol of the Comprehensive intelligent Hypertension managEment SyStem (CHESS) evaluation study assessing the effect of a multifaceted hypertension management system for supporting patients and physicians on BP lowering in primary care settings. MATERIALS AND METHODS: The CHESS evaluation study is a parallel-group, cluster-randomized controlled trial conducted in primary care settings in China. Forty-one primary care sites from 3 counties of China are randomly assigned to either the usual care or the intervention group with the implementation of the CHESS system, more than 1,600 patients aged 35 to 80 years with uncontrolled hypertension and access to a smartphone by themselves or relatives are recruited into the study and followed up for 12 months. In the intervention group, participants receive patient-tailored reminders and alerts via messages or intelligent voice calls triggered by uploaded home blood pressure monitoring data and participants' characteristics, while physicians receive guideline-based prescription instructions according to updated individual data from each visit, and administrators receive auto-renewed feedback of hypertension management performance from the data analysis platform. The multiple components of the CHESS system can work synergistically and have undergone rigorous development and pilot evaluation using a theory-informed approach. The primary outcome is the mean change in 24-hour ambulatory systolic BP from baseline to 12 months. DISCUSSION: The CHESS trial will provide evidence and novel insight into the effectiveness and feasibility of an implementation strategy using a comprehensive digital BP management system for reducing hypertension burden in primary care settings. TRIAL REGISTRATION: https://www. CLINICALTRIALS: gov, NCT05605418.

Research on Foot Contact State Detection Technology of Wheel-Legged Robot.

The accurate perception of external environment information through the robot foot is crucial for the mobile robot to evaluate its ability to traverse terrain. Adequate foot-end contact signals can provide robust support for robot motion control and decision-making processes. The shape and uncertain rotation of the wheel-legged robot foot end represent a significant challenge to sensing the robot foot-end contact state, which current foot-end sensing schemes cannot solve. This paper presents a sensing method for the tire stress field of wheel-legged robots. A finite element analysis was conducted to study the deformation characteristics of the foot-end tire under force. Based on this analysis, a heuristic contact position estimator was designed that utilizes symmetrical deformation characteristics. Strain sensors, arranged in an array, extract the deformation information on the inner surface of the tire at a frequency of 200 Hz. The contact position estimator reduces the dimensionality of the data and fits the eigenvalues to the estimated contact position. Using support vector regression, the force estimator utilizes the estimated contact position and sensor signal to estimate the normal reaction force, designated as FZ. The sensing system is capable of detecting the contact position on the wheel circumference (with a root mean square error of 1.150°), as well as the normal force of 160 N on the Z axis (with a root mean square error of 6.04%). To validate the efficacy of the sensor detection method, a series of randomized and repeated experiments were conducted on a self-constructed test platform. This novel approach offers a promising avenue for perceiving contact states in wheel-legged robots.

Harnessing Catalytic RNA Circuits for Construction of Artificial Signaling Pathways in Mammalian Cells.

Engineering of genetic networks with artificial signaling pathways (ASPs) can reprogram cellular responses and phenotypes under different circumstances for a variety of diagnostic and therapeutic purposes. However, construction of ASPs between originally independent endogenous genes in mammalian cells is highly challenging. Here we report an amplifiable RNA circuit that can theoretically build regulatory connections between any endogenous genes in mammalian cells. We harness the system of catalytic hairpin assembly with combination of controllable CRISPR-Cas9 function to transduce the signals from distinct messenger RNA expression of trigger genes into manipulation of target genes. Through introduction of these RNA-based genetic circuits, mammalian cells are endowed with autonomous capabilities to sense the changes of RNA expression either induced by ligand stimuli or from various cell types and control the cellular responses and fates via apoptosis-related ASPs. Our design provides a generalized platform for construction of ASPs inside the genetic networks of mammalian cells based on differentiated RNA expression.

Association of Leisure-Time Physical Activity and Mortality Risk in High Cardiovascular Risk Population with and without Left Ventricular Hypertrophy.

Background: Increased leisure-time physical activity (LTPA) is linked with decreased mortality risk, while also with increased left ventricular mass, which may induce left ventricular hypertrophy (LVH). We investigated whether LVH modifies the association between higher LTPA and lower mortality risk in population at high cardiovascular risk. Methods: In a prospective national cohort, we used the left ventricular mass/body surface area (LVM/BSA) method to define LVH. Baseline LTPA was self-reported and divided into: low ( < 500 metabolic equivalent of task [MET]) min/week), moderate (500-1999 MET min/week) and high ( ≥ 2000 MET-min/week). Analyses of the dose-response relationship between LTPA and left ventricular mass were performed using restricted cubic spline regression. A multivariate adjusted Cox proportional hazards regression analysis was used to estimate hazard ratios (HRs). Results: A total of 163,006 participants (55.3% females, mean [standard deviation] age, 62.4 [7.4] years) were included. During a median of 4.8 years of follow-up, 6586 (4.0%) died from all causes and 3024 (1.9%) from cardiovascular causes. Multivariate adjusted Cox proportional hazards regression analyses revealed that moderate and high LTPA were linked with less cardiovascular and all-cause mortality risk than low LTPA in the absence of LVH. In those with LVH, the association of high (0.83, 0.69-0.99) or moderate (0.72, 0.56-0.91) LTPA with cardiovascular mortality risk persisted. For all-cause mortality risk, this association was only significant in high LTPA (0.73, 0.61-0.86), while marginal in moderate LTPA (0.96, 0.84 to 1.08). Overall, the correlation patterns between LTPA and mortality risk appears distinct between those with LVH and those without LVH; the modification of LVH was not significant regarding mortality risk among the high cardiovascular risk population (all-cause: p-value for interaction = 0.074; cardiovascular cause: p-value for interaction = 0.581), except in females regarding all-cause mortality risk (p-value for interaction = 0.006). Conclusions: The association between higher LTPA and lower mortality risk was not modified by LVH in high cardiovascular risk population. However, the presence of LVH altered this association in females regarding the all-cause mortality risk.

Time-Varying Effect of Physical Activity on Mortality Among Myocardial Infarction Survivors: A Nationwide Population-Based Cohort Study.

Background: Physical activity (PA) is an important component of secondary prevention after myocardial infarction (MI). The mortality risk of MI survivors varies at different post-MI periods, yet the time-varying effect of total PA is unclear. We aimed to investigate the association between different volumes and patterns of total PA and mortality at different post-MI periods. Methods: Using data from the China Patient-centered Evaluative Assessment of Cardiac Events Million Persons Project, we divided the screened MI survivors into within-1-year and beyond-1-year groups based on the duration between their baseline interview and MI onset. Total PA was divided into insufficient ( < 3000 metabolic equivalent of task [MET] minutes/week) and sufficient PA. Sufficient PA was further categorized as moderate and high (3000-4500 and > 4500 MET minutes/week) volumes; leisure ( ≥ 50%) and non-leisure ( > 50%) patterns. Data on mortality were derived from the National Mortality Surveillance System and Vital Registration of the Chinese Center for Disease Control and Prevention. Cox proportional hazard models were fitted to estimate hazard ratios (HRs) and 95% confidence intervals (CIs). Restricted cubic spline regression analyses were performed to examine the dose-response association between PA and mortality. Results: During the follow-up (median 3.7 years) of the 20,653 post-MI patients, 751 patients died. In the within-1-year group, moderate (HR: 0.59, 95% CI: 0.40 to 0.88) and high (0.63, 0.45 to 0.88) volumes and both patterns (leisure: 0.52, 0.29 to 0.94; non-leisure: 0.64, 0.46 to 0.88) of PA were all associated with significantly lower risk of mortality, compared with insufficient PA. In the beyond-1-year group, the association was observed in high volume (0.69, 0.56 to 0.86) and both patterns (leisure: 0.64, 0.48 to 0.87; non-leisure: 0.79, 0.65 to 0.97). A non-linear relationship between PA and mortality was found in the within-1-year group (p for non-linearity < 0.001), while a linear relationship was demonstrated in the beyond-1-year group (p for non-linearity = 0.107). Conclusions: Sufficient total PA was associated with mortality risk reduction after MI, either leisure or non-leisure pattern. Different dose-response associations between PA and mortality were found at different post-MI periods. These results could promote individualized and scientifically derived secondary prevention strategies for MI.

Autophagy-amplifying nanoparticles evoke immunogenic cell death combined with anti-PD-1/PD-L1 for residual tumors immunotherapy after RFA.

Incomplete radiofrequency ablation (IRFA) triggers mild protective autophagy in residual tumor cells and results in an immunosuppressive microenvironment. This accelerates the recurrence of residual tumors and causes resistance to anti-PD-1/PDL1 therapy, which bringing a great clinical challenge in residual tumors immunotherapy. Mild autophagy activation can promote cancer cell survival while further amplification of autophagy contributes to immunogenic cell death (ICD). To this regard, we constructed active targeting zeolitic imidazolate framework-8 (ZIF-8) nanoparticles (NPs) loaded with STF62247 or both STF62247 and BMS202, namely STF62247@ZIF-8/PEG-FA (SZP) or STF62247-BMS202@ZIF-8/PEG-FA (SBZP) NPs. We found that SZP NPs inhibited proliferation and stimulated apoptosis of residual tumor cells exposed to sublethal heat stress in an autophagy-dependent manner. Further results discovered that SZP NPs could amplify autophagy in residual tumor cells and evoke their ICD, which dramatically boosted the maturation of dendritic cells (DCs). Through vaccination experiments, we found for the first time that vaccination with heat + SZP treatment could efficiently suppress the growth of new tumors and establish long-term immunological memory. Furthermore, SBZP NPs could remarkably promote the ICD of residual tumor cells, obviously activate the anti-tumor immune microenvironment, and significantly inhibit the growth of residual tumors. Thus, amplified autophagy coupled with anti-PD-1/PDL1 therapy is potentially a novel strategy for treating residual tumors after IRFA.

Effects of morphological characteristics of patent foramen ovale by transesophageal echocardiography on minimalist transcatheter closure in southern China.

OBJECTIVES: Transcatheter closure has become one of the main treatment methods for patent foramen ovale (PFO). However, the population in southern China is generally thin and the size of PFO is small, so the application of minimalist surgery is challenging. This study aimed to analyze the morphological characteristics of PFOs in southern China by transesophageal echocardiography (TEE), and to explore the influence on minimalist transcatheter closure. METHODS: About 110 patients with PFO closure in our hospital were selected. All cases were examined by TEE including the PFO size, length, septum secundum thickness, color characteristic and surrounding structures, and morphologically classified. During the operation, the procedure time, number of times for the guidewire attempting to pass the interatrial septum and the success rate of simply using J guidewire to cross the interatrial septum were recorded. RESULTS: About 110 cases of PFO were classified into two categories and four subtypes, including 55 cases with Uniform Channel Type (UCT, 50.0%), 16 cases with Irregular Channel Type (ICT, 14.6%), 15 cases with Right Funnel Type (RFT, 13.6%), and 24 cases with Left Funnel Type (LFT, 21.8%). According to the complexity of the procedure, they were divided into simple procedure (n = 73) and complex procedure (n = 37). Multivariate logistic regression showed that the anatomical types of PFO, the tunnel entrance size, and the tunnel entrance size <2 mm were independent factors affecting the complexity of procedure [OR = 2.819, 95% CI (1.124, 7.066), p = .027; OR = .027, 95% CI (.004, .208), p = .001; OR = 4.715, 95% CI (1.028, 21.619), p = .046]. With ICT and LFT groups, the procedure duration was relatively long (p < .001), number of times for the guidewire attempting to pass the interatrial septum was significantly increased (p < .001), and the success rate of simply using J guidewire to cross the interatrial septum was relatively low (p < .001). CONCLUSIONS: The PFO size in southern China was relatively small and characterized by large tunnel tension. It was concluded that TEE could clearly show the morphological characteristics of PFO, which could provide guidance for making more reasonable surgical plans in clinical practice, shorten the procedure time and improve the success rate of PFO closure.

Acute and 13 weeks subchronic toxicological evaluation of the flavonoid-rich extract of Sophora flavescens.

The roots of Sophora flavescens have a long history of use in Chinese medicine for the treatment of various medical conditions. Flavonoids from the ethyl acetate extract of S. flavescens have shown anti-inflammatory, anticancer, and antidiabetic properties. The objective of this study was to evaluate the toxicological profile of a flavonoid-rich extract of S. flavescens (SFEA). We conducted acute and sub-chronic oral toxicity studies of SFEA in Kunming (KM) mice and Sprague-Dawley (SD) rats. Acute oral administration of 9.0 g/kg SFEA did not result in mortality, clinical signs of toxicity, or abnormal changes in the body weight or food consumption patterns. No significant changes in hematological, blood biochemical, or histopathological parameters were observed. A 13-week sub-chronic toxicity study was conducted in SD rats; the rats were orally administrated with various doses of SFEA (in mg/kg): 0 (control), 40, 80, 400, 800, and 1200. Mortality, clinical signs, or treatment-related changes in body weight, food consumption, hematological parameters, blood biochemical parameters, organ weights, or histopathological parameters were not observed. We found that SFEA is practically nontoxic to KM mice at a dose of 9.0 g/kg and that the no-observed-adverse-effect-level (NOAEL) of SFEA in SD rats is greater than 1200 mg/kg.

Identification and profiling of microRNAs involved in the regenerative involution of mammary gland.

Regenerative involution is important for the subsequent lactation, but molecular mechanism has not been revealed. The crucial miRNA in tissue development indicates that miRNAs might participate in regenerative involution. In the present study, the mammary tissues of the dairy goats (n = 3) were collected via biopsy at wk-8 (time to dry off), -6, -4, -1, and + 1 relative to lambing for the Hematoxylin and Eosin staining and miRNA sequencing. Alveolar structures collapsed during regenerative involution, but the structures remained intact and distended. Among the 50 miRNA expression trajectories categorized by short time-series expression miner, two significant patterns were clustered. The differentially expressed miRNAs in the two patterns were mainly related to the self-renewal of tissue and enriched in pathways containing vesical-mediated transport, tissue development, tube development, vasculature development and epithelial development. The identification of the miRNAs will help in elucidating their regulatory roles in mammary gland development.

Using elastography-based multilayer perceptron model to evaluate renal fibrosis in chronic kidney disease.

BACKGROUND: Given its progressive deterioration in the clinical course, noninvasive assessment and risk stratification for the severity of renal fibrosis in chronic kidney disease (CKD) are required. We aimed to develop and validate an end-to-end multilayer perceptron (MLP) model for assessing renal fibrosis in CKD patients based on real-time two-dimensional shear wave elastography (2D-SWE) and clinical variables. METHODS: From April 2019 to December 2021, a total of 162 patients with CKD who underwent a kidney biopsy and 2D-SWE examination were included in this single-center, cross-sectional, and prospective clinical study. 2D-SWE was performed to measure the right renal cortex stiffness, and the corresponding elastic values were recorded. Patients were categorized into two groups according to their histopathological results: mild and moderate-severe renal fibrosis. The patients were randomly divided into a training cohort (n = 114) or a test cohort (n = 48). The MLP classifier using a machine learning algorithm was used to construct a diagnostic model incorporating elastic values with clinical features. Discrimination, calibration, and clinical utility were used to appraise the performance of the established MLP model in the training and test sets, respectively. RESULTS: The developed MLP model demonstrated good calibration and discrimination in both the training [area under the receiver operating characteristic curve (AUC) = 0.93; 95% confidence interval (CI) = 0.88 to 0.98] and test cohorts [AUC = 0.86; 95% CI = 0.75 to 0.97]. A decision curve analysis and a clinical impact curve also showed that the MLP model had a positive clinical impact and relatively few negative effects. CONCLUSIONS: The proposed MLP model exhibited the satisfactory performance in identifying the individualized risk of moderate-severe renal fibrosis in patients with CKD, which is potentially helpful for clinical management and treatment decision-making.

Preparation and Stability of Monodisperse Amorphous CaCO3 Microspheres of High Hardness and Sphericity.

Using amorphous CaCO3 (ACC) to biomimic the crustacean exoskeleton and optimize the physical and chemical properties of the polymeric phase of ACC holds great promise. Controlling the ACC morphology and stability is key in this process. For this article, monodisperse ACC microspheres, with a high sphericity of 0.973 ± 0.001 and a hardness of 0.6755 GPa, were prepared using the gas diffusion method in the presence of Mg2+. Their hardness is 3.58-16 times greater than that ever reported before for ACC microspheres. The stability of ACC is strongly affected by environmental conditions. The liquid phase and high temperature are not conducive to its stability, but ACC microspheres do have high stability under ambient conditions. After 100 days under such conditions, only a small amount of crystallization occurs, and their spherical shape survives intact. This article provides guidance for the preparation of ACC biomimetic composites, sheds light on the biological function of ACC in crustacean exoskeletons, and improves the theoretical understanding of the mechanism of biomineralization.

Alcohol use disorder and its association with quality of life and mortality in Chinese male adults: a population-based cohort study.

AIMS: We aimed to demonstrate the distribution of alcohol use disorder (AUD) in China and assess its association with quality of life and mortality. METHODS: We studied 367 120 men aged 35-75 years from 31 provinces in the China Patient-Centered Evaluative Assessment of Cardiac Events (PEACE) Million Persons Project. At baseline, AUD was assessed by alcohol use disorders identification test score, and EQ-5D-3L questionnaire was used to measure the quality of life. Mortality data was collected via National Mortality Surveillance System and Vital Registration. Mixed models were fitted to assess the associations of AUD with quality of life, and Cox proportional hazard models were fitted for the associations with all-cause and cause-specific mortality. RESULTS: We identified 39 163 men with AUD, which accounted for 10.7% of male participants and 25.8% of male drinkers. In the multivariable analysis, male drinkers who were aged 45-54 years, with higher education level, currently smoking, obese, with diagnosed hypertension, and without diagnosed cardiovascular diseases had higher rates of AUD. Male drinkers with AUD were less likely to have optimal QOL compared with those without AUD (OR: 0.63, 95% CI: 0.61-0.65, P < 0.001). Moreover, among male drinkers, AUD was prospectively associated with a 20% higher risk for all-cause mortality and a 30% higher risk for mortality from cancer. CONCLUSIONS: In China one fourth of men who drank alcohol had AUD, which was associated with lower QOL and higher risk of all-cause mortality. National policies or strategies are urgently needed to control alcohol-related harms.

Healthy lifestyle behaviours and all-cause and cardiovascular mortality among 0.9 million Chinese adults.

BACKGROUND: Healthy lifestyle behaviours are effective means to reduce the burden of diseases. This study was aimed to fill the knowledge gaps on the distribution, associated factors, and potential health benefits on mortality of four healthy lifestyle behaviours in China. METHODS: During 2015-2019, participants aged 35-75 years from 31 provinces were recruited by the China PEACE Million Persons Project. Four healthy lifestyle behaviours were investigated in our study, including non-smoking, none or moderate alcohol use, sufficient leisure time physical activity (LTPA), and healthy diet. RESULTS: Among 903,499 participants, 74.1% were non-smokers, 96.0% had none or moderate alcohol use, 23.6% had sufficient LTPA, 11.1% had healthy diet, and only 2.8% had all the four healthy lifestyle behaviours. The adherence varied across seven regions; the highest median of county-level adherence to all the four healthy lifestyle behaviours was in North China (3.3%) while the lowest in the Southwest (0.8%) (p < 0.05). Participants who were female, elder, non-farmers, urban residents, with higher income or education, hypertensive or diabetic, or with a cardiovascular disease (CVD) history were more likely to adhere to all the four healthy lifestyle behaviours (p < 0.001). County-level per capital Gross Domestic Product (GDP) was positively associated with sufficient LTPA (p < 0.05 for both rural and urban areas) and healthy diet (p < 0.01 for urban areas), while negatively associated with none or moderate alcohol use (p < 0.01 for rural areas). Average annual temperature was negatively associated with none or moderate alcohol use (p < 0.05 for rural areas) and healthy diet (p < 0.001 for rural areas). Those adhering to all the four healthy lifestyle behaviours had lower risks of all-cause mortality (HR 0.64 [95% CI: 0.52, 0.79]) and cardiovascular mortality (HR 0.53 [0.37, 0.76]) after a median follow-up of 2.4 years. CONCLUSIONS: Adherence to healthy lifestyle behaviours in China was far from ideal. Targeted health promotion strategies were urgently needed.

Penpolonin A-E, cytotoxic α-pyrone derivatives from Penicillium polonicum.

Five new α-pyrone derivatives, named penpolonin A-E (1-5), together with two known compounds (6-7) were acquired from the endophytic fungus Penicillium polonicum isolated from the roots of Camptotheca acuminata Decne. Their structures were established by combination of NMR and HRESIMS data and the absolute configurations of 1-5 were determined by NMR calculations and comparison of experimental and calculated ECD data. Compounds 3 and 7 exhibited moderate cytotoxicity against Hep-2, TU212 human laryngeal cancer cells with IC50 values ranging from 31.6 to 45.1 µg/ml, compound 4 showed weak cytotoxicity against the Hep-2 and TU212 cell lines with IC50 values of 69.2 and 68.7 µg/ml.

Genome-wide analysis of DNA methylation and risk of cardiovascular disease in a Chinese population.

BACKGROUND: Systemic studies of association of genome-wide DNA methylated sites with cardiovascular disease (CVD) in prospective cohorts are lacking. Our aim was to identify DNA methylation sites associated with the risk of CVD and further investigate their potential predictive value in CVD development for high-risk subjects. METHODS: We performed an epigenome-wide association study (EWAS) to identify CpGs related to CVD development in a Chinese population.We adopted a nested case-control design based on data from China PEACE Million Persons Project. A total of 83 cases who developed CVD events during follow-up and 83 controls who were matched with cases by age, sex, BMI, ethnicity, medications treatment and behavior risk factors were included in the discovery stage. Genome-wide DNA methylation from whole blood was detected using Infinium Human Methylation EPIC Beadchip (850 K). For significant CpGs [FDR(false discovery rate) < 0.005], we further validated in an independent cohort including 38 cases and 38 controls. RESULTS: In discovery set, we identified 8 significant CpGs (FDR < 0.005) associated with the risk of CVD after adjustment for cell components, demographic and cardiac risk factors and the first 5 principal components. Two of these identified CpGs (cg06901278 and cg09306458 in UACA) were replicated in another independent set (p < 0.05). Enrichment analysis in 787 individual genes from 1036 CpGs in discovery set revealed a significant enrichment for anatomical structure homeostasis as well as regulation of vesicle-mediated transport. Receiver operating characteristic (ROC) analysis showed that the model combined 8 CVD-related CpGs with baseline characteristics showed much better predictive effect for CVD occurrence compared with the model with baseline characteristics only [AUC (area under the curve) = 0.967, 95% CI (0.942 - 0.991); AUC = 0.621, 95% CI (0.536 - 0.706); p = 9.716E-15]. CONCLUSIONS: Our study identified the novel CpGs associated with CVD development and revealed their additional predictive power in the risk of CVD for high-risk subjects.

New flavonoids from the roots of Sophora davidii (Franch.) Skeels and their glucose transporter 4 translocation activities.

Five new flavanones, davidones A-E (1-5), one new isoflavonoid, cyclolicoisoflavones A3 (8), together with seven known compounds were isolated from the petroleum ether and the ethyl acetate fractions of the roots of Sophora davidii (Franch.) Skeels. The structures of new compounds were established by 1D and 2D NMR and MS data. The absolute configuration of 1-5 was assigned by NMR calculations and comparing its experimental and calculated ECD spectra. Flavanones were the main active principles responsible for the glucose transporter 4 (GLUT-4) translocation activities of SD-PE and SD-EtOAc. Compounds 1-7 and acacetin (12) promoted GLUT-4 translocation by the range of 1.35-3.00 folds, respectively.

Davidone C Induces the Death of Hepatocellular Carcinoma Cells by Promoting Apoptosis and Autophagy.

Davidone C is a newly discovered flavonoid compound purified from the ethyl acetate-soluble fraction of Sophora davidii (Franch.) Skeels. This study explored the anti-tumor activity of davidone C on hepatocellular carcinoma HepG2 and Bel-7402 cells and its mechanism through MTT method, morphological observation, flow cytometry and Western blotting. The results showed that davidone C significantly inhibited the proliferation of HepG2 and Bel-7402 cells in a time- and dose-dependent manner. The morphological changes of apoptotic cells can be observed under an inverted microscope, such as cell floating, chromosome condensation, apoptotic bodies, and other phenomena. The expressions of Bax, cleaved caspase-9, cleaved caspase-3 and cleaved PARP increased with the increase of dosage while Bcl-2 decreased, suggesting that the apoptotic mechanism might be related to the mitochondrial apoptotic pathway. Moreover, davidone C administration can down-regulate the expression of Grp78, and simultaneously up-regulate the expression of caspase-7 and caspase-12, indicating that the apoptotic mechanism might be related to the ERS pathway. In addition, davidone C can down-regulate the expression of p62, and simultaneously up-regulate the expression of LC3-I and LC3-II with a quantitative dependence, suggesting that the mechanism of apoptosis may be related to the autophagy signal pathway. All these results showed davidone C has potential effects on hepatocellular carcinoma.