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Abstract CRISPR/Cas12a is a highly promising detection tool. However, detecting single nucleotide variations (SNVs) remains challenging. Here, we elucidate Cas12a specificity through crRNA engineering and profiling of single- and double-base mismatch tolerance across three targets. Our findings indicate that Cas12a specificity depends on the number, type, location, and distance of mismatches within the R-loop. We also find that introducing a wobble base pair at position 14 of the R-loop does not affect the free energy change when the spacer length is truncated to 17 bp. Therefore, we develop a new universal specificity enhancement strategy via iterative crRNA design, involving truncated spacers and a wobble base pair at position 14 of the R-loop, which tremendously increases specificity without sacrificing sensitivity. Additionally, we construct a PAM-free one-pot detection platform for SARS-CoV-2 variants, which effectively distinguishes SNV targets across various GC contents. In summary, our work reveals new insights into the specificity mechanism of Cas12a and demonstrates significant potential for in vitro diagnostics.

Abstract Aims The endoscopic treatment of esophagogastric varices is challenging, and the nationwide application of endoscopic therapies for various types of esophagogastric varices and different clinical scenarios remains unclear. This study investigated the use of endoscopic therapy for portal hypertension in China. Methods This study used a questionnaire survey initiated by the Liver Health Consortium in China to investigate the use of endoscopic therapies for portal hypertension. Questionnaires were released online from January 30, 2023 to February 28, 2023 and filled out by chief physicians or senior instructors responsible for endoscopic therapies in participating hospitals across 31 provinces (autonomous regions and municipalities) in China. Comparisons of guideline adherence between primary and referral medical centers were performed using the chi‐square test or Fisher's exact test. Results In total, 836 hospitals participated in the survey. For primary and secondary prophylaxis of esophagogastric variceal bleeding (EGVB), adherence to the national guidelines was 72.5% (606/836) and 39.2% (328/836), respectively. Significant differences were observed in the rate of adherence between the primary and referral centers for primary (79.9% [111/139] vs. 71.0% [495/697], p = 0.033) and secondary prophylaxis (27.3% [38/139] vs. 41.6% [290/697], p = 0.002). Of the hospitals, 78.2% (654/836) preferred endoscopic therapies for acute EGVB, and the timing of endoscopy was usually within 12 h (48.5%, 317/654) and 12–24 h (36.9%, 241/654) after bleeding. Endoscopic therapy was more likely to be the first choice of treatment for acute EGVB in referral centers than in primary centers (82.6% [576/697] vs. 56.1% [78/139], p

Objectives: This study aimed to assess the knowledge, attitudes, and practices (KAP) and the training requirements of pediatric intensive care nurses regarding intra-abdominal pressure (IAP) monitoring, in order to provide a reference for the development of relevant training programs and operational procedures in clinical practice. Methods: This descriptive cross-sectional survey was conducted from April 2023 to June 2023. A convenience sample was created by recruiting 212 pediatric intensive care nurses in eight hospitals in Zhejiang Province. A self-developed IAP monitoring KAP assessment tool was used for evaluation, which included knowledge (14 items), attitude (6 items), and practice (8 items), three dimensions, 28 items. Results: The overall KAP score was 60.73 ± 8.35; the knowledge score was 7.84 ± 2.35, with a scoring rate of 56.0%; the attitude score was 25.16 ± 3.23, with a scoring rate of 83.9%; and the practice score was 28.44 ± 6.46, with a scoring rate of 69.3%. Nurses wh-o have received IAP monitoring training have higher KAP score, knowledge score and practice score than those who have not received it (P 30 (P

Abstract Diabetic nephropathy (DN) is a frequent and costly complication of diabetes with limited understandings of mechanisms and therapies. Emerging evidence points to the important roles of interleukin‐33 (IL‐33) in acute kidney injury, yet its contribution to DN is still unclear. We here found a ubiquitous increase of IL‐33 and its receptor (ST2) in murine models and patients with DN. Surprisingly, both IL‐33 and ST2 knockdown aggravated renal lesions in DN, while overexpression of IL‐33 also exacerbated the condition. Further population‐based analyses revealed a positive correlation of IL‐33 expression with renal dysfunction in DN patients. Individuals with high IL‐33 expression‐related polygenic risk score had a higher DN risk. These findings confirmed the harmful effects of IL‐33 on DN. Conversely, endogenous and exogenous partial reduction of IL‐33 signaling conferred renoprotective effects in vivo and in vitro. Mechanistically, IL‐33 induced senescence by regulating cell cycle factors in HK‐2 cells, and accordingly senescence led to renal cell damage through the secretion of senescence‐related secretory phenotype (SASP) including IL‐33 and prostaglandins. Together, elevated IL‐33 accelerates cellular senescence to drive DN possibly by SASP production, while a partial blockage improves renal injury and senescence. Our findings pinpoint a possible and new avenue for DN interventions.

The widespread adoption of arc additive manufacturing techniques across various industries has advanced the field of SS316L stainless steel manufacturing. It is crucial to acknowledge that different welding modes exert distinct influences on the forming and mechanical performance. This study analyzed the thermal input associated with four specific welding modes in LORCH MIG welding, clarifying the transition dynamics of molten droplets through waveform analysis and examining the resultant effects on microstructure and performance characteristics. The Pulse, Speed-Pulse-XT, and Twin-Pulse modes were found to induce spatter during the manufacturing process, consequently reducing molding efficiency in comparison to the SA-XT mode. Notably, the Twin-Pulse mode, characterized by double-pulse agitation, generated fish scale patterns along the lateral surfaces of the fabricated parts, promoting anisotropic grain growth. This microstructural refinement, compared to single-pulse samples with equivalent thermal input, resulted in enhanced mechanical properties. Nevertheless, the horizontal tensile strength of the three pulse modes was lower than the industrial standard for SA-XT mode and forging. In contrast, the SA-XT mode with an average hardness of 168.1 ± 6.9 HV and a tensile strength of 443.58 ± 5.7 MPa. Therefore, while three pulse modes offer certain microstructural advantages, the SA-XT mode demonstrates superior overall performance.

Accurate battery models are of great significance for the optimization design and management of lithium-ion batteries. This study uses a pseudo-two-dimensional electrochemical model combined with a three-dimensional thermal model to describe the electrodynamics and thermodynamics of commercial LIBs and adopts the concept of variable solid-state diffusion in the electrochemical model to improve the fitting ability of the model. Compared with the discharge curve without the VSSD concept, the progressiveness of the model is verified. On the other hand, by comparing the temperature distribution of batteries with different negative electrode thicknesses, it is found that the battery temperature decreases with the increase in battery thickness. At the same time, with the increase in active material volume fraction, the gradient of electrochemical performance is greater, and the heat generation rate is higher. This model can be used for online management of batteries, such as estimating charging status and internal temperature, and further constructing a lithium battery electrochemical capacity degradation model based on the VSSD concept to study the aging behavior of lithium batteries.

Abstract Background Few multi-ethnic dietary culture-sensitive food frequency questionnaires (FFQ) have been developed due to the complexity and diversity of cooking methods and styles. This study aimed to develop and validate a specific FFQ among multi-ethnic groups in Northwest China. Methods In the reliability study, 139 participants aged 20–65 completed two identical FFQs separated by 3 months. The relative validation of the FFQ was assessed by three 24-h recalls (24HR) employed in the interval of two FFQs, as a reference. Stratified analyses were also conducted by the major ethnic groups (Han nationality or Ethnic minority). Results For reproducibility, the median (range) of Spearman’s correlation coefficients (SCC) was 0.71 (0.43–0.84) for nutrients. The intra-class correlation coefficients (ICC) covered a spectrum from 0.39 to 0.78 (median: 0.64). Meanwhile, the weighted kappa values ranged from 0.11 to 0.64. For validity, the median (range) of Pearson’s correlation coefficients derived from the energy unadjusted and the adjusted values between FFQ and 24HR were 0.61 (0.12–0.79) and 0.56 (0.12–0.77), respectively. The results of correlation coefficients were similar between the two ethnic groups. Moreover, the Bland–Altman plots likewise demonstrated a satisfactory level of agreement between the two methods. Conclusions The FFQ showed acceptable reproducibility and moderate relative validity for evaluating dietary intake among multi-ethnic groups in northwest China. It could be a credible nutritional screening tool for forthcoming epidemiological surveys of these populations.

In this study, we analyzed the arc magnetic field to assess the stability of the arc welding process, particularly in robotic welding where direct measurement of welding current is challenging, such as under water. The characteristics of the magnetic field were evaluated based on low-frequency fluctuations and the symmetry of the signals. We used double-wire pulsed MIG welding for our experiments, employing Q235 steel with an 8.0 mm thickness as the material. Key parameters included an average voltage of 19.8 V, current of 120 A, and a wire feeding speed of 3.3 m/min. Our spectral analysis revealed significant correlations between welding stability and factors such as the direct current (DC) component and the peak power spectral density (PSD) frequency. To quantify this relationship, we introduced a novel approach using sample entropy and mix sample entropy (MSE) as new evaluation metrics. This method achieved a notable accuracy of 88%, demonstrating its effectiveness in assessing the stability of the robotic welding process.

Wire arc additive manufacturing (WAAM) technology enables the fabrication of functionally graded materials (FGMs) by adjusting the wire feed speed of different welding wires in a layer-by-layer manner. This study aimed to produce SS 316L/Inconel 625 FGMs with varying transition compositions using dual-wire arc additive manufacturing (D-WAAM). An optimization strategy for transition gradients was implemented to exclude component regions that are prone to defect formation (notably cracking), as well as to retain other component regions, thereby enhancing the overall mechanical properties of FGMs. The study revealed grain boundary cracking and demonstrated the lowest microhardness and tensile properties within a 20 wt.% Inconel 625 transition gradient zone, which negatively impacts the overall mechanical properties of FGMs. Then, as the content of Inconel 625 in the first transition region increased, cracks disappeared, microhardness increased and better tensile properties were obtained. The most optimal mechanical properties were enriched at 50 wt.% Inconel 625 content. In conclusion, the compositional gradient optimization strategy proves efficacious in eliminating component regions with poor mechanical properties and microdefects, ensuring excellent overall mechanical characteristics of FGMs.

Aluminum–magnesium (Al–Mg) alloys, known for their lightweight properties, are extensively utilized and crucial in the advancement of wire and arc additive manufacturing (WAAM) for direct high-quality printing—a focal point in additive manufacturing research. This study employed 1.2 mm ER5356 welding wire as the raw material to fabricate two sets of 30-layer thin-walled structures. These sets were manufactured using two distinct welding modes, speed-twin pulse (STP) and twin pulse (TP). Comparative evaluations of the surface quality, microstructures, and mechanical properties of the two sets of samples indicated that both the STP and TP modes were suitable for the WAAM of Al–Mg alloys. Analyses of grain growth in the melt pools of both sample sets revealed a non-preferential grain orientation, with a mixed arrangement of equiaxed and columnar grains. The STP mode notably achieved a refined surface finish, a reduced grain size, and a slight increase in tensile strength compared to the TP mode. From the comparison of the tensile data at the bottom, middle, and top of the two groups of samples, the additive manufacturing process in the STP mode was more stable.

BackgroundSalt substitutes are edible salts that contain less sodium chloride owing to its partial substitution with other minerals, which serve as an important and effective intervention and public health strategy targeting hypertension and its secondary diseases, despite a small degree of controversy. ObjectiveThis study aimed to identify the current salt substitute initiatives in various nations and intergovernmental organizations (IGOs) worldwide and summarize their types and characteristics. MethodsA scoping review was performed based on Arksey and O’Malley’s framework and the latest guidance from the Joanna Briggs Institute. Searches were conducted on Google; government websites on food, health, and other related topics; PubMed; Web of Science; and Google Scholar from January 2022 to May 2022. Initiatives related to salt substitutes that were included in the study focused on the involvement of governments or IGOs through the publication of standards, actions, collaborations, funding, and so on. Data were extracted into Microsoft Excel (version 2019; Microsoft Corp) based on predefined items and analyzed using narrative synthesis and frequency count methods. ResultsA total of 35 initiatives from 11 countries (of which 9 are high-income countries) and 3 IGOs were identified. We classified all salt substitute initiatives into 5 types, namely benefit-risk assessments and cautions; plans and actions; regulations and standards; labels; and food reformulation, cooperation with the food industry, and media. More than half (18/35, 51%) of the salt substitute initiatives were launched within the past 5 years. Except for regulations and standards, salt substitute initiatives are, in general, part of the salt reduction framework. No nation or IGO has yet reported on the monitoring and implications of the use of salt substitutes. ConclusionsDespite the limited number of salt substitute initiatives worldwide at present, a review on the different types and characteristics of such initiatives could be helpful in providing a reference for policy makers and stakeholders. Given the great potential of salt substitutes in improving hypertension and stroke, we call on more nations to pay attention to these substitutes and propose salt substitute initiatives in line with their national conditions.

In this work, polydopamine (PDA) was used to facilitate the reduction of silver nitrate (AgNO3) to silver nanoparticles (AgNPs) and the simultaneous loading on waste silk fabrics (PDA-SF/AgNPs). The removal activity of the obtained composite PDA-SF/AgNPs on different dyes in the presence of hydrogen peroxide (H2O2) and potassium peroxymonosulfate (PMS) was compared. The effects of reaction temperature, H2O2/PMS dosage, dye concentration, composite dosage, electrolyte concentration and time in two catalyst systems on the dye removal were discussed. The results showed the dye removal of Telon Red A2R reached 81% and 95% in 30–50 min in PDA-SF/AgNPs-H2O2 and PDA-SF/AgNPs-PMS system, respectively. Quenching experiments demonstrated that ·OH, ·O2– and SO4-· were produced during the degradation process, and the possible degradation pathway of Telon Red A2R were analyzed by HPLC-MS. In addition, the kinetic study indicated that the removal of dye followed quasi-second-order adsorption kinetics. These findings suggest that fabric-based composites have great potential and applications in catalysis and treatment of wastewater.

Abstract Background Oral administration offered a painless way and improved compliance for diabetics. However, the emerging GLP-1 analog peptide drugs for diabetes primarily rely on the injection route, and the development of oral dosage forms was hampered by the low oral bioavailability due to the structural vulnerability to digestive enzymes and molecule impermeability in the gastrointestinal tract. Results In this study, the non-covalent interaction between cholic acid (CA) and liraglutide (LIRA) was found and theoretically explained by molecular docking simulation. Formation of this physical complex of liraglutide and cholic acid (LIRA/CA Complex) reduced the self-aggregation of LIRA and accelerated intestinal epithelium penetration. By the anti-solvent method, LIRA/CA Complex was loaded into zein/rhamnolipids nanoparticles (LIRA/CA@Zein/RLs) with a loading efficiency of 76.8%. LIRA was protected from fast enzymatic degradation by the hydrophobic zein component. Meanwhile, Rhamnolipids, a glycolipid with surface activity, promoted endocytosis while also stabilizing the nanoparticles. The two components worked synergistically to ensure the delivery of LIRA/CA Complex to intestinal villi and improved oral absorption without disrupting tight junctions. LIRA/CA@Zein/RLs demonstrated a considerable intestinal epithelium absorption in mouse gastrointestinal section and a retention in vivo over 24 h, resulting in a significant and long-lasting hypoglycemic effect in Type 2 diabetes mice. Conclusion This study provided a promising oral delivery approach for LIRA and exhibited the potential for further translation into clinical application.

Abstract No fully validated risk-stratification strategies have been established in China where colonoscopies resources are limited. We aimed to develop and validate a fecal immunochemical test (FIT)-based risk-stratification model for colorectal neoplasia (CN); 10,164 individuals were recruited from 175 centers nationwide and were randomly allocated to the derivation (n = 6776) or validation cohort (n = 3388). Multivariate logistic analyses were performed to develop the National Colorectal Polyp Care (NCPC) score, which formed the risk-stratification model along with FIT. The NCPC score was developed from eight independent predicting factors and divided into three levels: low risk (LR 0–14), intermediate risk (IR 15–17), and high risk (HR 18–28). Individuals with IR or HR of NCPC score or FIT+ were classified as increased-risk individuals in the risk-stratification model and were recommended for colonoscopy. The IR/HR of NCPC score showed a higher prevalence of CNs (21.8%/32.8% vs. 11.0%, P

Abstract Background Osteoarthritis (OA) is a worldwide public health concern, mainly afflicting older adults. Although the etiology of OA remains unclear, environmental factors are increasingly considered as non-negligible risk factors. This study aims to evaluate the associations of urinary metals with OA risk and the mediated effect of biological aging. Methods Nine urinary metal concentrations were detected among 12,584 U.S. adults based on the National Health and Nutrition Examination Survey (NHANES), including barium (Ba), cadmium (Cd), cobalt (Co), cesium (Cs), molybdenum (Mo), lead (Pb), antimony (Sb), thallium (Tl), and uranium (Tu). Multivariable logistic regression and weighted quantile sum (WQS) regression were used to explore the associations of single metal and mixed metals with OA risk, respectively. Furthermore, biological aging was measured from different perspectives, including cell senescence (telomere length) and whole-body aging (phenotypic age and biological age). Mediation analyses were conducted to investigate the mediated effects of aging on the associations of metals with OA risk. Results In the single-exposure model, Cd, Co, and Cs were identified to be positively associated with OA risk, with odds ratios (OR) ranging from 1.48 to 1.64 (all P

Abstract Background Isoproterenol (ISO), a synthetic on selective β-adrenergic agonist, provides a simple and non-invasive method for inducing myocardial injury with lower mortality and higher reproducibility. Phlegm-damp syndrome, as known as “Tanshi” in Chinese, is one of Traditional Chinese Medicine (TCM) syndrome differentiation, which plays an important role in the development of cardiovascular diseases. However, the underlying mechanism remains unknown. Methods In our present study, a myocardial injury mouse model was introduced by ISO administration combined with high temperature and high humidity and high-fat diet to simulate phlegm-damp syndrome. Nontargeted metabolomics with LC–MS/MS was adopted to reveal serum metabolism profile for elucidating the possible molecular mechanism. Results The results of our study showed that phlegm-damp syndrome promoted ISO-induced myocardial injury by aggravating left ventricular hypertrophy and fibrosis, and increasing cardiac index. Our study also confirmed the presence of specific metabolites and disturbed metabolic pathways by comparing ISO mice and Tanshi mice, mainly including glycerophospholipid metabolism, arginine–proline metabolism, and sphingolipid signaling pathway. The lysoPCs, PCs, SMs, Sphingosine, and L-Arginine were the main metabolites that showed a difference between ISO and Tanshi mice, which might be the result of the underlying mechanism in the promotion of ISO-induced myocardial injury in mice with high temperature and high humidity and high-fat diet. Conclusion Our current study provides new insights into contribution of metabolism disorder in promotion of ISO-induced myocardial injury in mice with high temperature and high humidity and high-fat diet, and new targets for clinical diagnosis and pharmacologic treatment of cardiovascular disease with phlegm-damp syndrome.

IntroductionXin-Li-Fang (XLF), a representative Chinese patent medicine, was derived from years of clinical experience by academician Chen Keji, and is widely used to treat chronic heart failure (CHF). However, there remains a lack of high-quality evidence to support clinical decision-making. Therefore, we designed a randomized controlled trial (RCT) to evaluate the efficacy and safety of XLF for CHF.Methods and designThis multicenter, double-blinded RCT will be conducted in China. 300 eligible participants will be randomly assigned to either an XLF group or a control group at a 1:1 ratio. Participants in the XLF group will receive XLF granules plus routine care, while those in the control group will receive placebo granules plus routine care. The study period is 26 weeks, including a 2-week run-in period, a 12-week treatment period, and a 12-week follow-up. The primary outcome is the proportion of patients whose serum NT-proBNP decreased by more than 30%. The secondary outcomes include quality of life, the NYHA classification evaluation, 6-min walking test, TCM symptom evaluations, echocardiography parameters, and clinical events (including hospitalization for worsening heart failure, all-cause death, and other major cardiovascular events).DiscussionThe results of the study are expected to provide evidence of high methodological and reporting quality on the efficacy and safety of XLF for CHF.Clinical trial registrationChinese Clinical Trial Registration Center (www.chictr.org.cn). The trial was registered on 13 April 2022 (ChiCTR2200058649).

In the context of ageing, the combination of medical and health-care has become a major trend, and the introduction of the PPP model into combined medical and healthcare projects can solve the problem of lack of funds for the development of combined medical and health-care projects. Based on the evolutionary game theory, this paper constructs a three-party evolutionary game model to study the strategy choice of each party in the game process, and also analyses the evolutionary equilibrium strategy and the impact of parameter adjustment on the evolutionary strategy under different situations. The results show that the optimal solution is for the private sector to choose to provide high quality services, for the government to choose to actively regulate and for the public to choose to actively monitor. For each player, the incentives and disincentives influence their behavioural strategies. Based on the above analysis, this paper suggests establishing an independent regulator, broadening public feedback channels, and improving PPP-related laws, as well as innovating project operation methods and improving enterprise operational capabilities.

Abstract Whether the definition of hypertension according to 2017 AHA/ACC guidelines and blood pressure (BP) changes was related to the increased risk of chronic kidney disease (CKD) remained debated. This prospective cohort study aimed to investigate the association of BP and long‐term BP change with CKD risk with different glucose metabolism according to the new hypertension guidelines. This study examined 12 951 participants and 11 183 participants derived from the older people cohort study, respectively. Participants were divided into three groups based on blood glucose and the risks were assessmented by the logistic regression model. During a 10 years of follow‐up period, 2727 individuals developed CKD (21.1%). Compared with those with BP

This research investigated the effect of ultrasound (US) pretreatment prior to spray drying on the powder flow and moisture sorption behaviour of micellar casein concentrate (MCC). MCC produced from skim milk microfiltration was sonicated at energy intensity of 0 (control), 47 J/mL (S-2000), 62 J/mL (S-3000) and 76 J/mL (S-4000). The results revealed that US pretreatment significantly increased the average particle size (D50) from 82.46 μm to 100.73 μm and reduced the surface fat content from 19.2% to 13.8%, resulting in decreased basic flow energy, cake energy and cohesion. Besides, the US treated samples showed relatively poor ability to acquire the moisture from the atmosphere than the control. Protein structure analysis showed that α-helix decreased with enhanced US power, while β-sheet and surface hydrophobicity increased, implying hydrophobic groups were exposed and water sorption rate was impeded. As a result, US pretreatment can improve the powder flow and potentially reduce the negative effect of cake formation at high humidity.

Cardiovascular diseases, the notorious killer, are mainly caused by atherosclerosis (AS) characterized by lipids, cholesterol, and iron overload in plaques. Macrophages are effector cells and accumulate to the damaged and inflamed sites of arteries to internalize native and chemically modified lipoproteins to transform them into cholesterol-loaded foam cells. Foam cell formation is determined by the capacity of phagocytosis, migration, scavenging, and the features of phenotypes. Macrophages are diverse, and the subsets and functions are controlled by their surrounding microenvironment. Generally, macrophages are divided into classically activated (M1) and alternatively activated (M2). Recently, intraplaque macrophage phenotypes are recognized by the stimulation of CXCL4 (M4), oxidized phospholipids (Mox), hemoglobin/haptoglobin complexes [HA-mac/M(Hb)], and heme (Mhem). The pro-atherogenic or anti-atherosclerotic phenotypes of macrophages decide the progression of AS. Besides, apoptosis, necrosis, ferroptosis, autophagy and pyrotopsis determine plaque formation and cardiovascular vulnerability, which may be associated with macrophage polarization phenotypes. In this review, we first summarize the three most popular hypotheses for AS and find the common key factors for further discussion. Secondly, we discuss the factors affecting macrophage polarization and five types of macrophage death in AS progression, especially ferroptosis. A comprehensive understanding of the cellular and molecular mechanisms of plaque formation is conducive to disentangling the candidate targets of macrophage-targeting therapies for clinical intervention at various stages of AS.