De novo synthesis of a novel hapten and development of a monoclonal antibody-based immunoassay for the detection of dichlorvos and trichlorfon.

The absence of high-affinity antibodies has hindered the development of satisfactory immunoassays for dichlorvos (DDVP) and trichlorfon (TCP), two highly toxic organophosphorus pesticides. Herein, the de novo synthesis of a novel anti-DDVP hapten was introduced. Subsequently, a specific anti-DDVP monoclonal antibody (Mab) was produced with satisfying affinity to DDVP (IC50: 12.4 ng mL-1). This Mab was highly specific to DDVP, and TCP could readily convert into DDVP under mild alkaline conditions. Leveraging this insight, an indirect competitive ELISA was successfully developed for simultaneous detection of DDVP and TCP. The limit of detection in rice, cabbage and apple for DDVP /TCP was found to be 12.1/14.6 µg kg-1, 7.3/8.8 µg kg-1 and 6.9/8.3 µg kg-1, respectively. This study not only provides an effective strategy for producing a high-quality anti-DDVP Mab but also affords a reliable and cost-effective tool suitable for high-throughput detection of DDVP and TCP in food samples.

Loading rutin on surfaces by the layer-by-layer assembly technique to improve the oxidation resistance and osteogenesis of titanium implants in osteoporotic rats.

The purpose of this study was to construct a rutin-controlled release system on the surface of Ti substrates and investigate its effects on osteogenesis and osseointegration on the surface of implants. The base layer, polyethylenimine (PEI), was immobilised on a titanium substrate. Then, hyaluronic acid (HA)/chitosan (CS)-rutin (RT) multilayer films were assembled on the PEI using layer-by-layer (LBL) assembly technology. We used scanning electron microscopy (SEM), Fourier transform infrared (FTIR) spectroscopy and contact angle measurements to examine all Ti samples. The drug release test of rutin was also carried out to detect the slow-release performance. The osteogenic abilities of the samples were evaluated by experiments on an osteoporosis rat model and MC3T3-E1 cells. The results (SEM, FTIR and contact angle measurements) all confirmed that the PEI substrate layer and HA/CS-RT multilayer film were effectively immobilised on titanium. The drug release test revealed that a rutin controlled release mechanism had been successfully established. Furthermore, thein vitrodata revealed that osteoblasts on the coated titanium matrix had greater adhesion, proliferation, and differentiation capacity than the osteoblasts on the pure titanium surface. When MC3T3-E1 cells were exposed to H2O2-induced oxidative stressin vitro, cell-based tests revealed great tolerance and increased osteogenic potential on HA/CS-RT substrates. We also found that the HA/CS-RT coating significantly increased the new bone mass around the implant. The LBL-deposited HA/CS-RT multilayer coating on the titanium base surface established an excellent rutin-controlled release system, which significantly improved osseointegration and promoted osteogenesis under oxidative stress conditions, suggesting a new implant therapy strategy for patients with osteoporosis.

Radix Rehmanniae Praeparata promoted zebrafish fin regeneration through aryl hydrocarbon receptor-dependent autophagy.

HEADINGS ETHNOPHARMACOLOGICAL RELEVANCE: Rehmanniae Radix Praeparata (RRP), a staple in traditional Chinese medicine, is derived from Rehmannia glutinosa Libosch and is renowned for its wound-healing properties. Despite its clinical prevalence, the molecular mechanisms underlying RRP's wound-healing effects have not been fully elucidated. AIM OF THE STUDY: This research endeavored to delineate the molecular and cellular mechanisms underlying the beneficial effects of RRP on wound healing, utilizing a zebrafish model. MATERIALS AND METHODS: Zebrafish larvae at 3 days post-fertilization were amputated at the fin and subsequently treated with RRP. The pro-wound healing and regenerative effects of RRP were evaluated through morphological analysis, assessment of cell proliferation and apoptosis, Additionally, mechanistic insights were gained through a comprehensive approach encompassing network pharmacology analysis, cell tracing, RNA-sequencing, CRISPR/Cas9 gene editing, and pharmacological inhibition. RESULTS: Our findings demonstrate that RRP significantly accelerates caudal fin regeneration in zebrafish following injury by suppressing cell apoptosis, promoting cell proliferation, and upregulating the expression of regenerative-related genes. Furthermore, RRP triggers autophagy signals during the regenerative process, which is attenuated by the autophagy inhibitor chloroquine (CQ). Notably, the administration of RRP enhances the expression of ahr1 and ahr2 in the regenerating fin. Genetic knockout of ahr1a, ahr1b, or ahr2 using CRISPR/Cas9, or pharmacological blockade of AHR signals with the antagonist CH-223191, diminishes the regenerative potential of RRP. Remarkably, zebrafish lacking ahr2 completely lose their fin regeneration ability. Additionally, inhibition of AHR signaling suppresses autophagy signaling during fin regeneration. CONCLUSIONS: This study uncovers that RRP stimulates fin regeneration in zebrafish by inducing AHR signals and, at least partially, activating the autophagy process. These findings provide novel insights into the molecular mechanisms underlying the wound-healing effects of RRP and may pave the way for the development of novel therapeutic strategies.

LiveBoost: A GB-based prediction system for liver fibrosis in chronic hepatitis B patients in China - A multi-center retrospective study.

Background: The aim of this study was to evaluate the accuracy of LiveBoost™, a gradient boosting (GB)-based prediction system based on standard biochemical values (AST, ALT, platelet count) and age, in Chinese patients with chronic hepatitis B (CHB) and compare its performance with FIB-4 (fibrosis-4 score) and APRI (the aspartate transaminase to platelet ratio index). Methods: This retrospective trial enrolled 454 participants, including 279 CHB patients who underwent liver biopsy and 175 normal controls from 3 centers in China. All participants underwent laboratory blood testing. LiveBoost was constructed using GB and FIB-4 and APRI were calculated from laboratory data. Results: LiveBoost outperformed APRI and FIB-4 in predicting hepatic fibrosis and cirrhosis. The GB model had an AUROC of 0.977 for CHB diagnosis, 0.804 for early and advanced fibrosis, and 0.836 for non-cirrhosis and cirrhosis, compared to AUROC of 0.554, 0.673 and 0.720 for FIB-4, AUROC of 0.977, 0.652 and 0.654 for APRI. Conclusions: LiveBoost is a more reliable and cost-effective method than APRI and FIB-4 for assessing liver fibrosis in Chinese patients with CHB.

5-(3',4'-dihydroxyphenyl)-γ-valerolactone, a microbiota metabolite of flavan-3-ols, activates SIRT1-mediated autophagy to attenuate H2O2-induced inhibition of osteoblast differentiation in MC3T3-E1 cells.

Phenolic compounds are promising agents for the prevention of osteoporosis. 5-(3',4'-dihydroxyphenyl)-γ-valerolactone (DHPV) is the major microbiota metabolite of the flavan-3-ols phenolic compound. Herein, we aimed to investigate the potential mechanisms underlying the effects of DHPV on an osteoblast cell model with H2O2-induced oxidative injury. The MC3T3-E1 cell cultured with H2O2 was used as an oxidative injury model after pretreating with DHPV. Pretreatment with DHPV significantly attenuated cell viability decline, enhanced the activity of alkaline phosphatase and mineralization capacity in MC3T3-E1 cells. Reduced reactive oxygen species (ROS) and malondialdehyde (MDA) levels as well as increased in mitochondrial membrane potential and superoxide dismutase (SOD) activities indicated that DHPV affected both the oxidative and antioxidative processes in the cells. DHPV administration increased the LC3-II/I ratio and Beclin-1 protein levels, thereby promoting autophagy, which perhaps contributes to ROS elimination. However, the inhibition of Sirtuin 1 (SIRT1) by SIRT1 small interfering RNA reduced the protective effect of DHPV or SRT1720, as revealed by the increased ROS and MDA levels and decreased SOD, LC3-II/I ratio and Beclin-1 levels. DHPV may promote autophagy and reduce oxidative stress through the SIRT1-mediated pathway, thereby protecting MC3T3-E1 cells from H2O2-induced oxidative damage.

Associations of plasma retinol and α-tocopherol levels with skeletal muscle health in Chinese children.

Childhood is a critical period for muscle accumulation. Studies in elders have reported that antioxidant vitamins could improve muscle health. However, limited studies have assessed such associations in children. This study included 243 boys and 183 girls. A seventy-nine-item FFQ was used to investigate dietary nutrients intake. Plasma levels of retinol and α-tocopherol were measured using high-performance liquid chromatography with MS. Dual X-ray absorptiometry was used to assess appendicular skeletal muscle mass (ASM) and total body fat. The ASM index (ASMI) and ASMI Z-score were then calculated. Hand grip strength was measured using a Jamar® Plus+ Hand Dynamometer. Fully adjusted multiple linear regression models showed that for each unit increase in plasma retinol content, ASM, ASMI, left HGS and ASMI Z-score increased by 2·43 × 10-3 kg, 1·33 × 10-3 kg/m2, 3·72 × 10-3 kg and 2·45 × 10-3 in girls, respectively (P < 0·001-0·050). ANCOVA revealed a dose-response relationship between tertiles of plasma retinol level and muscle indicators (Ptrend: 0·001-0·007). The percentage differences between the top and bottom tertiles were 8·38 %, 6·26 %, 13·2 %, 12·1 % and 116 % for ASM, ASMI, left HGS, right HGS and ASMI Z-score in girls, respectively (Pdiff: 0·005-0·020). No such associations were observed in boys. Plasma α-tocopherol levels were not correlated with muscle indicators in either sex. In conclusion, high circulating retinol levels are positively associated with muscle mass and strength in school-age girls.

Development a high-throughput zebrafish embryo acute toxicity testing method based on OECD TG 236.

The Organization for Economic Co-operation and Development (OECD）Test Guideline (TG) 236 for zebrafish embryo acute toxicity testing was adopted for chemical toxicity assessment in 2013. Due to the increasing demand for prediction and evaluation of the acute toxicity using zebrafish embryos, we developed a method based on OECD 236 test guideline with the aim to improve the testing efficiency. We used 4-128 cell stage zebrafish embryos and performed an exposure assay in a 96-well microtiter plate, observing the lethality endpoints of embryos at 48-h postexposure. A total of 32 chemicals (two batches) were used in the comparison study. Our results indicated that the logarithmic LC50 (half lethal concentration) obtained by the modified method exhibited good correlation with that obtained by the OECD 236 testing method, and the R2 of the linear regression analysis was 0.9717 (0.9621 and 0.9936 for the two batches, respectively). Additionally, the intra- and inter-laboratory coefficient of variation (CVs) for the LC50 from the testing chemicals (17 chemicals in second batch) was less than 30%, except for CuSO4. Therefore, the developed method was less time-consuming and demonstrated a higher throughput for toxicity testing compared to the prior method. We argue the developed method could be used as an additional choice for high-throughput zebrafish embryo acute toxicity test.

Association of the gut microbiota and fecal short-chain fatty acids with skeletal muscle mass and strength in children.

We aimed to investigate whether the gut microbiota and fecal short-chain fatty acids (SCFAs) are associated with skeletal muscle mass and strength in healthy Chinese children aged 6-9 years. In this study, 412 children were enrolled. 16S rRNA gene sequencing was used to characterize the gut microbiota compositions. Fecal SCFAs were quantified using high-performance liquid chromatography. Dual X-ray absorptiometry was used to measure the total body lean soft tissue mass (TSM), total body fat mass (TBF), appendicular skeletal muscle mass (ASM), and appendicular fat mass (AFM). TSM/height2 (TSMI), ASM/height2 (ASMI), TSM/weight (TSMR), ASM/weight (ASMR), and the ratio of TSM/TBF and ASM/AFM were calculated. Handgrip strength (HGS) was measured using the Jamar® Plus+ Hand Dynamometer. A multiple regression analysis after adjustment for covariates and multiple test correction showed some operational taxonomic units in partial least squares models identified by Multivariate methods with Unbiased Variable selection analysis such as genera of Faecalibacterium, Lachnospira, Lachnospiraceae_ND3007_group, and Lachnospiraceae_UCG-004 were positively correlated with at least one measure of TSM, TSMI, ASM, ASMI, and ASMI Z-score (ß: 0.103-0.143, pFDR : .008-.032) but negatively correlated with at least one measure of TSMR, TSM/TBF, ASMR, ASM/AFM, and ASMR Z-score (ß: -0.185 to 0.124, pFDR  = .008-.045). Children with higher fecal butyric acid, acetic acid, and total SCFA levels exhibited higher TSM, ASM, TSMI, ASMI, and ASMI Z-score and lower TSM/TBF, ASM/AFM, TSMR, ASMR, and ASMR Z-score. However, after additional adjustment for TBF or body mass index, only the associations for Faecalitalea and Pyramidobacter still existed. Mediation analysis suggested that total body fat significantly mediated 66.3%-95.3% of the estimated association of microbiota and SCFAs with TSM, ASM, and ASMI Z-score. Our results suggest that the associations of gut microbiota and SCFAs with skeletal muscle quality in children may largely depend upon on total body fat content.

The associations of gut microbiota and fecal short-chain fatty acids with bone mass were largely mediated by weight status: a cross-sectional study.

PURPOSE: We aimed to investigate whether the gut microbiota and fecal short-chain fatty acids (SCFAs) are associated with bone mass in healthy children aged 6-9 years. METHODS: In this study, 236 healthy children including 145 boys and 91 girls were enrolled. 16S rRNA gene sequencing was used to characterize the composition of their gut microbiota. Total and 10 subtypes of SCFAs in the fecal samples were determined by high-performance liquid chromatography. Dual X-ray absorptiometry was used to measure the bone mineral density (BMD) and bone mineral content (BMC) for total body (TB) and total body less head (TBLH). Z score of TBLH BMD was calculated based on the recommended reference. RESULTS: Four gut microbiota principal components (PCs) were identified by the compositional principal component analysis at the genus level. After adjustment of covariates and controlling for the false discovery rate, multiple linear regression analysis showed that PC3 score (positive loadings on genera Lachnoclostridium and Blautia) was significantly negatively associated with TBLH BMD/BMC/Z score, TB BMC and pelvic BMD (ß: - 0.207 to - 0.108, p: 0.002-0.048), whereas fecal total and several subtypes of SCFAs were correlated positively with TBLH BMD/Z score and pelvic BMD (ß: 0.118-0.174, p: 0.038-0.048). However, these associations disappeared after additional adjustment for body weight. Mediation analysis suggested that body weight significantly mediated 60.4% and 78.0% of the estimated association of PC3 score and SCFAs with TBLH BMD Z score, respectively. CONCLUSIONS: The associations of gut microbiota composition and fecal SCFA concentrations with bone mass in children were largely mediated by body weight.

Investigation of the Ballistic Performance of GFRP Laminate under 150 m/s High-Velocity Impact: Simulation and Experiment.

The ballistic resistance of GFRP laminates subjected to high-velocity impact was studied. Based on the damage situation of GFRP laminate observed from the single-stage gas gun testing, the three-dimensional (3D) model combining strain rate effect and Hashin failure criterion was established, and the result presented good agreement between the simulation and experiment. Three factors, including layer angle, stacking sequence and proportion of different layer angles, were taken into consideration in the models. An orthogonal test method was used for the analysis, which can reduce the number of simulations effectively without sacrificing the accuracy of the result. The result indicated a correlation between the ballistic resistance and layouts of GFRP laminates, on which the stacking sequence contributed stronger influence. What was more, the laminate with layer angles 0°/90° and ±45° presented greater ballistic resistance than the other angle pairs, and adopting an equal proportion of different layer angles is helpful for GFRP laminates to resist impact as well.

Associations of Plasma Copper, Magnesium, and Calcium Levels with Blood Pressure in Children: a Cross-sectional Study.

Studies of the relationship of copper (Cu), magnesium (Mg), and calcium (Ca) levels with blood pressure in children are limited. This cross-sectional study included 443 children aged 6-9 years from Guangzhou, China. Plasma concentrations of Cu, Mg, and Ca were measured by inductively coupled plasma mass spectrometry. Systolic blood pressure (SBP) and diastolic blood pressure (DBP) were measured with an electronic sphygmomanometer. Elevated SBP and DBP were defined as the 90th percentile or greater (age and sex specific) of reference values for Chinese children. Abnormal blood pressure (ABP) was defined as an elevated SBP and/or DBP. The plasma Cu concentration was positively correlated with blood pressure, body mass index (BMI), and the C-reactive protein (CRP) level, whereas the plasma Mg and Ca concentrations were negatively correlated with SBP. A higher Cu concentration (T3 vs. T1) was associated with a higher risk of ABP (odds ratio (OR) 2.55, 95% confidence interval (CI) 1.20 to 5.40). In contrast, children in the top tertiles of Mg (OR 0.42, 95% CI 0.18 to 0.98) and Ca (OR 0.28, 95% CI 0.11 to 0.70) concentrations showed lower risks of ABP than those in the bottom tertiles. Path analysis showed that the CRP levels and BMI mediated the associations between the Cu concentration and ABP. We found that higher plasma Mg and Ca concentrations were negatively related to blood pressure in children aged 6-9 years. In contrast, a higher plasma Cu concentration was positively correlated with the risk of ABP, and the association was mediated by CRP and BMI.

Relationship Between Plasma Copper Concentration and Body Fat Distribution in Children in China: a Cross-Sectional Study.

This study aimed to explore the association of the plasma copper concentration with body fat distribution and the potential mediating effect of inflammation status in children. A total of 454 children were recruited in this study. Dual-energy X-ray absorptiometry was applied to measure the fat mass (FM) and fat mass percentages (FM%) at whole body, trunk, appendicular, android, and gynoid regions. Android to gynoid fat mass ratio and fat mass to lean mass (FM/LM) ratio at whole body, trunk, and appendicular sites were calculated. Plasma copper concentration was measured via inductively coupled plasma mass spectrometry. C-reactive protein (CRP) was determined by ELISA. After adjusting for covariates, multiple linear regression analyses showed that, for every additional unit increase in the plasma copper concentration, the FM, FM%, and FM/LM at whole body and subregions increased by 0.030-0.472 kg (P < 0.001-0.019), 0.013-1.04% (P = 0.007-0.042), and 0.021-0.030 (P < 0.001), respectively. Mediating analysis suggested that CRP significantly mediated 22.0-30.6% (P < 0.001) of the estimated association of copper with FM% and FM/LM at whole body and limbs. Thus, children with higher plasma levels of copper tended to have a higher regional and overall body fat deposition, and this relationship was partly mediated by inflammatory status.