Validating eDNA measurements of the richness and abundance of anurans at a large scale.

In some situations, environmental DNA (eDNA) metabarcoding is a more accurate tool for measuring species richness of a taxon than conventional methods. Whether this tool can reliably estimate the abundance of a taxon remains unresolved. We examined the reliability of metabarcoding for measuring anuran diversity compared to a commonly used traditional line transect method (TLTM) through the replicate sampling of three visits across 71 waterbodies (ponds or reservoirs) in Liuheng, China. We also investigated the relative contributions of species-specific characteristics and the physiochemical properties of a waterbody on the relative read count across species and waterbodies. We found that eDNA metabarcoding had a higher detection probability for each of seven anuran species found in the sampling region than TLTM. Furthermore, the relative read count estimated by metabarcoding was positively correlated with the density or relative density of individuals identified with the TLTM across waterbodies for every species. Species-specific characteristics of anurans, such as density, relative density, body mass, biomass and relative biomass, accounted for substantial variations in the read count across species and waterbodies, while physiochemical factors, including pH, temperature, water volume, vegetation and elevation, had little effect on the read count. Our results based on robust sampling suggest that metabarcoding enables more reliable and efficient measurements of anuran occurrence at a large scale during a short-term survey (within 15 days) than that obtained by the TLTM, and offers an alternative tool for quantifications of anuran abundance. Density or biomass is better and more reliable indicator of anuran abundance associated with read count than relative density or relative biomass.

Metabolism and disposition of corylifol A from Psoralea corylifolia: metabolite mapping, isozyme contribution, species differences and identification of efflux transporters for corylifol A-O-glucuronide in HeLa1A1 cells.

Corylifol A (CA), a phenolic compound from Psoralea corylifolia, possessed several biological properties but poor bioavailability. Here we aimed to investigate the roles of cytochromes P450s (CYPs), UDP-glucuronosyltransferases (UGTs) and efflux transporters in metabolism and disposition of CA.Metabolism of CA was evaluated in HLM, expressed CYPs and UGTs. Chemical inhibitors and shRNA-mediated gene silencing of multidrug resistance-associated proteins (MRPs) and breast cancer resistance protein (BCRP) were performed to assess the roles of transporters in CA disposition.Three oxidated metabolites (M1-M3) and two glucuronides (M4-M5) were detected. The intrinsic clearances (CLint) values of M1 and M4 in HLM were 48.10 and 184.03 µL/min/mg, respectively. Additionally, CYP1A1, 2C8 and 2C19 were identified as main contributors with CLint values of 13.01-49.36 µL/min/mg, while UGT1A1, 1A7, 1A8 and 1A9 were with CLint values ranging from 85.01 to 284.07 µL/min/mg. Furthermore, activity correlation analysis proved CYP2C8, UGT1A1 and 1A9 were the main active hepatic isozymes. Besides, rats and monkeys were appropriate model animals. Moreover, dipyridamole and MK571 both could significantly inhibit M4 efflux. Gene silencing results also indicated MRP4 and BCRP were major contributors in HeLa1A1 cells.Taken together, CYPs, UGTs, MRP4 and BCRP were important determinants of CA pharmacokinetics.

Environmental causes of between-population difference in growth rate of a high-altitude lizard.

BACKGROUND: Ectothermic animals living in cold (high latitude or high elevation) regions are predicted to grow slower due to limited thermal opportunities for activity and food resources than those living in warm regions. However, the Qinghai toad-headed lizards (Phrynocephalus vlangalii) grow faster and reach a larger adult size at a high-elevation site than at a low-elevation site. In this study, we aimed to identify the genetic and environmental causes of this between-population difference in growth rate by conducting mark-recapture and common garden experiments on juvenile growth rate, and investigating the thermal environment, lizard body temperature, potential prey availability at the two elevation sites. RESULTS: Compared with low-elevation individuals, high-elevation juvenile lizards had higher growth rates in the field, but grew at similar rates in the laboratory. High-elevation lizards had higher active body temperatures than low-elevation lizards despite similar air temperatures in the period of field investigation. The high-elevation site had relatively more and larger preys than the low-elevation site. CONCLUSIONS: Inter-population difference in growth rate of P. vlangalii may primarily result from developmental plasticity in response to the difference in environmental resources, rather than genetic differentiation. The higher growth rate of high-elevation lizards is likely associated with higher potential food availability and higher active body temperatures.

Resolving the complexity of vitellogenins and their receptors in the tetraploid Atlantic salmon (Salmo salar): Ancient origin of the phosvitin-less VtgC in chondrichthyean fishes.

Egg yolk proteins are mainly derived from vitellogenin (Vtg), and serve as essential nutrients during early development in oviparous organisms. Vertebrate Vtgs are predominantly synthesized in the liver of maturing females, and are internalized by the oocyte after binding to specific surface receptors (VtgR). Here, we clarify the evolutionary history of vertebrate Vtgs, including the teleost VtgC, which lacks phosvitin, and investigate the repertoire of Vtgs and VtgRs in the tetraploid Atlantic salmon (Salmo salar). Conserved synteny of the vtg genes in elephant fish (Callorhinchus milii) strongly indicates that the vtg gene cluster was present in the ancestor of tetrapods and ray-finned fish. The shortened phosvitin in the VtgC ortholog of this chondrichthyean fish may have resulted from early truncation events that eventually allowed the total disappearance of phosvitin in teleost VtgC. In contrast, the tandem-duplicated VtgCs identified in the spotted gar (Lepisosteus oculatus) both contain the phosvitin domain. The Atlantic salmon genome harbors four vtg genes encoding the complete VtgAsa1, phosvitin-less VtgC, and truncated VtgAsb proteins; vtgAsa2 is a pseudogene. The three vtg genes were mainly expressed in the liver of maturing females, and the vtgAsa1 transcript predominated prior to spawning. The splice variant lacking the O-linked sugar domain dominated ovarian expression of vtgr1 and vtgr2. Strongly increased vtgAsa1 expression during vitellogenesis contrasted with the peaks of vtgr1 and vtgr2 in the previtellogenic oocytes, which gradually decreased over the same period. Recycling of the oocyte VtgRs is probably not sufficient to maintain receptor number during vitellogenesis.

Research on preparation and related properties of macro-micro porous mullite ceramic skeletons via twice pore-forming technology.

A lot of solid waste coal gangue is produced every year in the process of coal mining and coal washing, which poses a great threat to human health. How to deal with coal gangue properly is still a serious problem. In this study the macro-micro composite porous mullite ceramic skeletons were successfully prepared using solid waste coal gangue and α-Al2O3 as main raw materials via twice pore-forming technology. The main phase composition of the porous ceramic skeletons was mullite tested by X-ray Diffractometer (XRD). The morphology and microstructure of the porous ceramic skeletons were analyzed by Scanning Electron Microscope (SEM). The results show that the microstructure of porous ceramic skeletons was mainly composed of mullite whiskers. With the increase of sintering temperature from 1200 °C to 1350 °C, the maximum length of mullite whiskers grew up from 2.68 µm to 8.10 µm and their average length grew up from 0.78 µm to 2.98 µm. The maximum compressive strength of the porous ceramic skeletons with 30 PPI and 45 PPI were 1.25 MPa and 1.54 MPa tested by Universal Testing Machine (UTM) at the sintering temperature of 1250 °C, respectively. The linear shrinkage, bulk density and pore stem density of the porous ceramic skeletons became larger with the rising of sintering temperatures from 1150 °C to 1350 °C. However, the corresponding performance values of 45 PPI porous ceramic skeletons were greater than that of 30 PPI at the same sintering temperature. The prepared porous ceramic skeletons will be used in ceramic-metal wear-resistant composites for the later research and the study provides a new idea for coal gangue on the comprehensive utilization with high added value and brings both good environmental and economic benefits.

Variations in Genetic Diversity of Invasive Species Lithobates catesbeianus in China.

The introduction and subsequent range expansion of the American bullfrog (Lithobates catesbeianus) is part of a rising trend of troublesome biological invasions happening in China. This detrimental amphibious invasive species has strong adaptability. After its introduction and spread, it established its own ecological niche in many provinces of China, and its range has continued to expand to more areas. Previous studies recorded the introduction time of bullfrogs and calculated the changes in their genetic diversity in China using mitochondria, but the specific introduction route in China is still unknown. Expanding upon previous research, we employed whole-genome scans (utilizing 2b-RAD genomic sequencing) to examine single nucleotide polymorphisms (SNPs) and microsatellites within Lithobates catesbeianus to screen the genomes of these invasive amphibian species from eight Chinese provinces and two U.S. states, including Kansas, where bullfrogs originate. A total of 1,336,475 single nucleotide polymorphic loci and 17 microsatellite loci were used to calculate the genetic diversity of bullfrogs and their migration pathways. Our results suggest that the population in Hunan was the first to be introduced and to spread, and there may have been multiple introductions of subpopulations. Additionally, the genetic diversity of both the SNP and microsatellite loci in the Chinese bullfrog population was lower than that of the US population due to bottleneck effects, but the bullfrogs can adapt and spread rapidly. This study will offer crucial insights for preventing and controlling future introductions into the natural habitats in China. Additionally, it will assist in devising more precise strategies to manage the existing populations and curtail their continued expansion, as well as aim to improve clarity and originality while mitigating plagiarism risk.

Antibiotic Resistance and Genetic Profiles of Vibrio parahaemolyticus Isolated from Farmed Pacific White Shrimp (Litopenaeus vannamei) in Ningde Regions.

To better understand the antibiotic resistance, virulence genes, and some related drug-resistance genes of Vibrio parahaemolyticus in farmed pacific white shrimp (Litopenaeus vannamei) in Ningde regions, Fujian province, we collected and isolated a total of 102 strains of V. parahaemolyticus from farmed pacific white shrimp in three different areas of Ningde in 2022. The Kirby-Bauer disk method was used to detect V. parahaemolyticus resistance to 22 antibiotics, and resistant genes (such as quinolones (qnrVC136, qnrVC457, qnrA), tetracyclines (tet A, tetM, tetB), sulfonamides (sulI, sulII, sulIII), aminoglycosides (strA, strB), phenicols (cat, optrA, floR, cfr), ß-lactams (carB), and macrolides (erm)) were detected by using PCR. The findings in this study revealed that V. parahaemolyticus was most resistant to sulfamoxazole, rifampicin, and erythromycin, with resistance rates of 56.9%, 36.3%, and 33.3%, respectively. Flufenicol, chloramphenicol, and ofloxacin susceptibility rates were 97.1%, 94.1%, and 92.2%, respectively. In all, 46% of the bacteria tested positive for multi-drug resistance. The virulence gene test revealed that all bacteria lacked the tdh and trh genes. Furthermore, 91.84% and 52.04% of the isolates were largely mediated by cat and sulII, respectively, with less than 5% resistance to aminoglycosides and macrolides. There was a clear mismatch between the antimicrobial resistance phenotypes and genotypes, indicating the complexities of V. parahaemolyticus resistance.

Risk factor analysis and nomogram for predicting gastroparesis in patients with type 2 diabetes mellitus.

Purpose: The incidence of gastroparesis is higher in individuals diagnosed with type 2 diabetes mellitus (T2DM) compared to the healthy individuals. Our study aimed to explore the risk factors for gastroparesis in T2DM and to establish a clinical prediction model (nomogram). Methods: Our study enlisted 694 patients with T2DM from two medical centers over a period of time. From January 2020 to December 2022, 347 and 149 patients were recruited from the Beilun branch of Zhejiang University's First Affiliated Hospital in the training and internal validation cohorts, respectively. The external validation cohort consisted of 198 patients who were enrolled at Nanchang University's First Affiliated Hospital from October 2020 to September 2021. We conducted univariate and multivariate logistic regression analyses to select the risk factors for gastroparesis in patients with T2DM; subsequently,we developed a nomogram model. The performance of the nomogram was evaluated using the area under the receiver operating characteristic (ROC) curve (AUC), calibration curve, and decision curve analysis(DCA). Results: Four clinical variables, including age, regular exercise, glycated hemoglobin level(HbA1c), and Helicobacter pylori (H. pylori) infection, were identified and included in the model. The model demonstrated excellent discrimination, with an AUC of 0.951 (95% CI = 0.925-0.978) in the training group, and 0.910 (95% CI = 0.859-0.961) and 0.875 (95% CI = 0.813-0.937) in the internal and external validation groups, respectively. The calibration curve showed good consistency between prediction of the model and observed gastroparesis. The DCA also demonstrated good clinical efficacy. Conclusion: The nomogram model developed in this study showed good performance in predicting gastroparesis in patients with T2DM.

eDNA Metabarcoding Reveals the Species-Area Relationship of Amphibians on the Zhoushan Archipelago.

The species-area relationship is important for understanding species diversity patterns at spatial scales, but few studies have examined the relationship using environmental DNA (eDNA) techniques. We investigated amphibian diversity on 21 islands of the Zhoushan Archipelago and nearby mainland areas in China using the combination of eDNA metabarcoding and the traditional line transect method (TLTM) and identified the species-area relationship for amphibians on the islands. The mean detection probability of eDNA is 0.54, while the mean detection probability of TLTM is 0.24. The eDNA metabarcoding detected eight amphibian species on the islands and nine species in the mainland areas, compared with seven species on the islands and nine species in the mainland areas that were identified by TLTM. Amphibian richness on the islands increased with island area and habitat diversity. The species-area relationship for amphibians in the archipelago was formulated as the power function (S = 0.47A0.21) or exponential function (S = 2.59 + 2.41 (logA)). Our results suggested that eDNA metabarcoding is more sensitive for the detection of amphibian species. The combined use of eDNA metabarcoding and the traditional line transect method may optimize the survey results for amphibians.

Effects of sodium intake, age, gender, blood sampling time on distribution of plasma aldosterone, renin activity, deoxycorticosterone, cortisol, cortisone, and 24 h urinary aldosterone levels in normotensive individuals based on LC-MS/MS.

PURPOSE: This study aims to analyze the distribution of plasma aldosterone, renin activity, deoxycorticosterone (DOC), cortisol, cortisone, and 24 h urinary aldosterone (24 h-uAld) levels based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) method. MATERIALS AND METHODS: Plasma and 24 h urine were collected from 129 healthy volunteers in Northeast China. The effect of sodium intake, age, gender, blood sampling time on plasma aldosterone concentration (PAC), plasma renin activity (PRA), PAC to PRA ratio (ARR), DOC, cortisol, cortisone, cortisol to cortisone ratio, and 24 h-uAld were investigated by nonparametric test, multiple linear regression and Harris-Boyd's standard deviate test. RESULTS: There was no significant difference observed in 24 h-uAld, PAC (AM), PRA(AM), ARR (AM), DOC (AM), cortisol (AM), cortisone (AM), and cortisol to cortisone (AM) between high and low sodium intake group. Significant differences were observed between morning and afternoon sampling groups in terms of PAC, ARR, DOC, cortisol, and cortisone. Reference intervals (RIs) of 24 h-uAld, PAC (AM) were recommended to be partitioned by gender. RI of PRA was recommended age stratification. CONCLUSION: We recommend that the same reference interval could be used regardless of sodium intake. Gender is the main influence factor for 24 h-uAld, PAC, and ARR. Age is key influence factor for PRA.

Pathogenic selection promotes adaptive immune variations against serious bottlenecks in early invasions of bullfrogs.

Adaptive genetic variations are key for understanding evolutionary processes influencing invasions. However, we have limited knowledge on how adaptive genetic diversity in invasive species responds to new pathogenic environments. Here, we compared variations in immune major histocompatibility complex (MHC) class-II ß gene and neutral loci in relation to pathogenic chytrid fungus (Batrachochytrium dendrobatidis, Bd) infection across invasive and native populations of American bullfrog between China and United States (US). Chinese invasive populations show a 60% reduction in neutral cytb variations relative to US native populations, and there were similar MHC variation and functional diversity between them. One MHC allele private to China was under recent positive selection and associated with decreased Bd infection, partly explaining the lower Bd prevalence for Chinese populations than for native US populations. These results suggest that pathogen-mediated selection favors adaptive MHC variations and functional diversity maintenance against serious bottlenecks during the early invasions (within 15 generations) of bullfrogs.

The role of closed-loop management of fever patients in a county-level medical community for the prevention and control of the post-coronavirus disease 2019 pandemic.

We built a closed-loop management model for patients with fever in a county-level medical community and explored the role of this model in post-coronavirus disease 2019 (COVID-19) epidemic prevention and control. The subjects included 83,791 patients with fever treated in designated hospitals between February 2020 and April 2021. A pre-hospital, in-hospital, and post-hospital management system for patients with fever in the county-level medical community was established to allow the closed-loop management of these patients. SPSS software (version 13.0) was used to analyze the methods of visiting the hospital, nucleic acid detection in the hospital, and location of the patients after the hospital visit. Chi-square tests were used to compare the methods of visiting and location after hospital visits between patients with and without an epidemiological history. The number of patients with fever in the fever clinic showed a logarithm change (R2 = 0.4710), accompanied by seasonal changes. The number of fever patients with an epidemiological history decreased logarithmically monthly (R2 = 0.8876). Among patients with fever, 99.64% sought medical treatment on their own, with relatively low proportions undergoing home quarantine and requiring centralized quarantine special vehicles. After visiting the fever clinics, 98.56% of patients isolated at home or were monitored, with small proportions of patients requiring hospital admission or centralized isolation. However, the proportions of patients with home and centralized isolation with epidemiology were relatively high, accounting for 20.55% and 27.40% of cases, respectively. Compared to the overall population of patients with fever, the difference was statistically significant (χ2 = 48.881, P = .000). The establishment of a closed-loop management model for patients with fever in a county-level medical community strengthened the management of these patients. No local cases occurred in Beilun District between March 2020 and April 2021. In the post-COVID-19 era, all medical institutions in the county-level medical community strengthened infectious disease pre-examination and triage and promoted the formation of a strategic pattern of initial diagnosis at the grassroots level, 2-way referral, upper and lower linkage, and joint epidemic prevention. This management was more conducive to COVID prevention and control by hierarchical management according to the presence or absence of an epidemiological history.

Preparation and Characterization of Multilayer pH-Responsive Hydrogel Loaded Ganoderma lucidum Peptides.

To develop a safe, targeted, and efficient assembly of a stable polypeptide delivery system, in this work, chitosan, sodium alginate, and sodium tripolyphosphate were used as materials for the preparation of hydrogels. M-SCT hydrogels were prepared by ionic gelation and the layer-by-layer (LBL) method. The composite hydrogels exhibited excellent pH sensitivity and Ganoderma lucidum peptides (GLP) loading capacity. The prepared hydrogels were characterized and evaluated. The internal three-dimensional network structure of the hydrogel was observed by scanning electron microscopy (SEM), and Fourier transform infrared (FT-IR) spectroscopy confirmed the electrostatic interactions among the components. X-ray diffraction (XRD) was used to observe the crystal structure of the hydrogel. The maximum peptide encapsulation efficiency was determined to be 81.73%. The digestion stability and thermal stability of M-SCT hydrogels loaded GLP were demonstrated to be improved. The amount of peptides released from the GLP/M-SCT-0.75 hydrogels in simulated gastric fluid was lower than 30%. In addition, the ABTS assays showed that the free radical scavenging ability of the GLP/M-SCT-0.75 hydrogels confirmed the efficacy of the hydrogels in retaining the antioxidant activity of GLP. The study suggested the M-SCT-0.75 hydrogels had a great deal of potential as a peptide carrier for oral delivery.

Peptides/receptors signaling during plant fertilization.

Double fertilization is a unique and particularly complicated process for the generation alternation of angiosperms. Sperm cells of angiosperms lose the motility compared with that of gymnosperms. The sperm cells are passively carried and transported by the pollen tube for a long journey before targeting the ovule. Two sperm cells are released at the cleft between the egg and the central cell and fused with two female gametes to produce a zygote and endosperm, respectively, to accomplish the so-called double fertilization process. In this process, extensive communication and interaction occur between the male (pollen or pollen tube) and the female (ovule). It is suggested that small peptides and receptor kinases play critical roles in orchestrating this cell-cell communication. Here, we illuminate the understanding of phases in the process, such as pollen-stigma recognition, the hydration and germination of pollen grains, the growth, guidance, and rupture of tubes, the release of sperm cells, and the fusion of gametes, by reviewing increasing data recently. The roles of peptides and receptor kinases in signaling mechanisms underlying cell-cell communication were focused on, and directions of future studies were perspected in this review.

Identification of naturally occurring inhibitors in Xian-Ling-Gu-Bao capsule against the glucuronidation of estrogens.

Xian-Ling-Gu-Bao (XLGB) capsule, a well-known traditional Chinese medicine prescription, is widely used for the treatment of osteoporosis. It could significantly increase the levels of estrogen in ovariectomized rats and mice. However, this working mechanism has not been well elucidated. Considering that UDP-glucuronosyltransferase (UGT) enzymes are the important enzymes that inactivate and regulate estrogen activity in vivo, this study aimed to identify the bioactive compounds from XLGB against the glucuronidation of estrogens. First, thirty compounds were considered as candidate bioactive compounds based on our previous studies including pharmacological evaluation, chemical profiles, and metabolic profiles. Second, the characteristics of estrogen glucuronidation by uridine diphosphate glucuronic acid (UDPGA)-supplemented human liver microsomes (HLM), human intestine microsomes (HIM), and expressed UGT enzymes were determined, and the incubation systems of their key UGT enzymes were optimized. Then, inhibitory effects and mechanisms of XLGB and its main compounds toward the key UGT isozymes were further investigated. As a result, estrogen underwent efficient glucuronidation by HLM and HIM. UGT1A10, 1A1, and 2B7 were mainly responsible for the glucuronidation of estrone, ß-estradiol, and estriol, respectively. For E1 and E2, UGT1A10 and 1A1 tended to mediate estrogen-3-O-glucuronidation, while UGT2B7 preferred catalyzing estrogen-16-O-glucuronidation. Furthermore, the incubation system for active UGT isoforms was optimized including Tris-HCl buffer, detergents, MgCl2 concentration, ß-glucuronidase inhibitors, UDPGA concentration, protein concentration, and incubation time. Based on optimal incubation conditions, eleven, nine, and nine compounds were identified as the potent inhibitors for UGT1A10, 1A1, and 2B7, respectively (IC50 < 4.97 µM and Ki < 3.35 µM). Among them, six compounds (bavachin, isobavachin, isobavachalcone, neobavaisoflavone, corylifol A, and icariside II) simultaneously demonstrated potent inhibitory effects against these three active enzymes. Prenylated flavanols from Epimedium brevicornu Maxim., prenylated flavonoids from Psoralea corylifolia L., and salvianolic acids from Salvia miltiorrhiza Bge. were characterized as the most important and effective compounds. The identification of potent natural inhibitors of XLGB against the glucuronidation of estrogen laid an important foundation for the pharmacodynamic material basis.

Effectiveness of a Multifaceted Intervention for Prevention of Obesity in Primary School Children in China: A Cluster Randomized Clinical Trial.

Importance: A rapid nutritional transition has caused greater childhood obesity prevalence in many countries, but the repertoire of effective preventive interventions remains limited. Objective: To determine the effectiveness of a novel multifaceted intervention for obesity prevention in primary school children. Design, Setting, and Participants: A cluster randomized clinical trial was conducted during a single school year (from September 11, 2018, to June 30, 2019) across 3 socioeconomically distinct regions in China according to a prespecified trial protocol. Twenty-four schools were randomly allocated (1:1) to the intervention or the control group, with 1392 eligible children aged 8 to 10 years participating. Data from the intent-to-treat population were analyzed from October 1 to December 31, 2019. Interventions: A multifaceted intervention targeted both children (promoting healthy diet and physical activity) and their environment (engaging schools and families to support children's behavioral changes). The intervention was novel in its strengthening of family involvement with the assistance of a smartphone app. The control schools engaged in their usual practices. Main Outcomes and Measures: The primary outcome was the change in body mass index (BMI; calculated as weight in kilograms divided by height in square meters) from baseline to the end of the trial. Secondary outcomes included changes in adiposity outcomes (eg, BMI z score, prevalence of obesity), blood pressure, physical activity and dietary behaviors, obesity-related knowledge, and physical fitness. Generalized linear mixed models were used in the analyses. Results: Among the 1392 participants (mean [SD] age, 9.6 [0.4] years; 717 boys [51.5%]; mean [SD] BMI, 18.6 [3.7]), 1362 (97.8%) with follow-up data were included in the analyses. From baseline to the end of the trial, the mean BMI decreased in the intervention group, whereas it increased in the control group; the mean between-group difference in BMI change was -0.46 (95% CI, -0.67 to -0.25; P < .001), which showed no evidence of difference across different regions, sexes, maternal education levels, and primary caregivers (parents vs nonparents). The prevalence of obesity decreased by 27.0% of the baseline figure (a relative decrease) in the intervention group, compared with 5.6% in the control group. The intervention also improved other adiposity outcomes, dietary, sedentary, and physical activity behaviors, and obesity-related knowledge, but it did not change moderate- to vigorous-intensity physical activity, physical fitness, or blood pressure. No adverse events were observed during the intervention. Conclusions and Relevance: The multifaceted intervention effectively reduced the mean BMI and obesity prevalence in primary school children across socioeconomically distinct regions in China, suggesting its potential for national scaling. Trial Registration: ClinicalTrials.gov Identifier: NCT03665857.

[Extraction technology of diester-type alkaloids from Radix Aconiti lateralis].

OBJECTIVE: To determine the suitable extraction technology of diester-type alkaloids, such as mesaconitine, aconitine and hypaconitine from the roots of Aconitum carmichaeli (Radix Aconiti lateralis). METHODS: The contents of mesaconitine, aconitine and hypaconitine were determined by HPLC. Single-factor experiment was used to study the extraction factors. RESULTS: The suitable extraction technology for diester-type alkaloids was as follows: coarse sizings of Radix Aconiti lateralis was extracted by 10 times ethanol for 3 times (each time for 1 day) at 15 degrees C. The total yield of diester-type alkaloids was 0.57%, including 0.16% aconitine, 0.032% mesaconitine and 0.38% hypaconitine, and the purity was 18.81%. CONCLUSION: The extraction technology is efficient, harmfulless, economical, convenient and can be used for industrial production.

Identification of an Exopolysaccharide Biosynthesis Gene in Bradyrhizobium diazoefficiens USDA110.

Exopolysaccharides (EPS) play critical roles in rhizobium-plant interactions. However, the EPS biosynthesis pathway in Bradyrhizobium diazoefficiens USDA110 remains elusive. Here we used transposon (Tn) mutagenesis with the aim to identify genetic elements required for EPS biosynthesis in B. diazoefficiens USDA110. Phenotypic screening of Tn5 insertion mutants grown on agar plates led to the identification of a mutant with a transposon insertion site in the blr2358 gene. This gene is predicted to encode a phosphor-glycosyltransferase that transfers a phosphosugar onto a polyprenol phosphate substrate. The disruption of the blr2358 gene resulted in defective EPS synthesis. Accordingly, the blr2358 mutant showed a reduced capacity to induce nodules and stimulate the growth of soybean plants. Glycosyltransferase genes related to blr2358 were found to be well conserved and widely distributed among strains of the Bradyrhizobium genus. In conclusion, our study resulted in identification of a gene involved in EPS biosynthesis and highlights the importance of EPS in the symbiotic interaction between USDA110 and soybeans.

Applications of electrochemical biosensors based on functional antibody-modified screen-printed electrodes: a review.

The detection of biomolecular analytes is of great importance in clinical, environmental, and argo-food areas, among which the electrochemical methodology is attracting much attention. In particular, screen-printed electrode (SPE)-based sensing applications have exhibited potential possibility for on-site detection, especially for fast clinical biomarker detection, since they provide a miniaturized but robust and portable electrode detection system. In this context, we focused on the modification of SPE with functional antibodies to improve the electrochemical detection performance in versatile sensing applications, particularly for COVID-19 detection. These antibodies were immobilized onto the electrode surface via various methodologies, through which the powerful potential from the modification of SPE was revealed. Finally, more novel and excellent works on the biomolecular modification of SPE and the prospects of this technology from its state-of-art status to commercialization are previewed and future perspectives in this field are mentioned.

Characterization of metabolic activity, isozyme contribution and species differences of bavachin, and identification of efflux transporters for bavachin-O-glucuronide in HeLa1A1 cells.

OBJECTIVES: Bavachin is a bioactive natural flavonoid with oestrogen-like activity. Here, we aimed to investigate its metabolic and disposal fates involving in CYPs, UGTs and efflux transporters. METHODS: Phase I metabolism and glucuronidation were performed by human liver microsomes (HLM). Reaction phenotyping and activity correlation analysis were performed to identify the main CYP and UGT isozymes. Chemical inhibition and gene knock-down approaches were employed to explore the function of BCRP and MRPs. KEY FINDINGS: Five phase I metabolites (M1-M5) and three glucuronides (G1-G3) were identified. The CLint values for M4 and G1 by HLM were 127.99 and 1159.07 µl/min per mg, respectively. Reaction phenotyping results suggested CYP1A1 (208.85 µl/min per mg) and CYP2C9 (107.51 µl/min per mg), and UGT1A1 (697.19 µl/min per mg), UGT1A7 (535.78 µl/min per mg), UGT1A8 (247.72 µl/min per mg) and UGT1A9 (783.68 µl/min per mg) all participated in the metabolism of bavachin. In addition, activity correlation analysis also supported the results above. Furthermore, the metabolism exhibited marked species differences, and rabbits were the appropriate model animals. Moreover, MRP4 was identified as the main contributor based on chemical inhibition and gene silencing approaches. CONCLUSIONS: CYP1A1 and CYP2C9, UGT1A1, UGT1A7, UGT1A8 and UGT1A9, and MRP4 all played important roles in the metabolism and disposition of bavachin.