An Optimized Microwave-Assisted Digestion Method to Analyze the Amino Acids Profile of Quisqualis Fructus from Different Planted Origins.

This study aims to establish a rapid and convenient microwave-assisted digestion method for sample pretreatment to determine amino acid profiles in natural products. This method was applied to analyze the amino acid profiles of Quisqualis Fructus (QF) from different planted origins. The microwave-assisted digestion conditions were optimized by a response surface methodology (RSM), and 17 amino acids in different planted origins of QF were determined by an automatic amino acid analyzer according to the optimized digestion conditions. The contents of 17 amino acids in QF from different planted origins were further analyzed by fingerprint and chemometric analysis. The temperature of microwave digestion at 167 °C, time of microwave digestion at 24 min, and a solid-liquid ratio of 46.5 g/mL was selected as the optimal digestion conditions. The total content of 17 amino acids in QF from different planted origins ranged from 71.88 to 91.03 mg/g. Amino acid composition and nutritional evaluation indicated that the content of medicinal amino acids was higher than aromatic amino acids. The results of fingerprint analysis reflected that the similarity between the 16 batches of QF ranged from 0.889 to 0.999, while chemometrics analysis indicated amino acid content in QF varied from different planted origins, and six important differential amino acids were screened. Compared with the traditional extraction method, microwave-assisted digestion with response surface optimized has the advantages of rapidity, convenience, and reliability, which could be used to study the amino acid profiles in natural products. The amino acid profile of QF indicated that it has a rich medicinal nutritional value. Different planted origins of QF have a high degree of similarity and could be effectively distinguished by chemometric analysis.

METTL3-mediated N6-methyladenosine modification of STAT5A promotes gastric cancer progression by regulating KLF4.

N6-methyladenosine (m6A) is the predominant post-transcriptional RNA modification in eukaryotes and plays a pivotal regulatory role in various aspects of RNA fate determination, such as mRNA stability, alternative splicing, and translation. Dysregulation of the critical m6A methyltransferase METTL3 is implicated in tumorigenesis and development. Here, this work showed that METTL3 is upregulated in gastric cancer tissues and is associated with poor prognosis. METTL3 methylates the A2318 site within the coding sequence (CDS) region of STAT5A. IGF2BP2 recognizes and binds METTL3-mediated m6A modification of STAT5A through its GXXG motif in the KH3 and KH4 domains, leading to increased stability of STAT5A mRNA. In addition, both METTL3 and IGF2BP2 are positively correlated with STAT5A in human gastric cancer tissue samples. Helicobacter pylori infection increased the expression level of METTL3 in gastric cancer cells, thereby leading to the upregulation of STAT5A. Functional studies indicated that STAT5A overexpression markedly enhances the proliferation and migration of GC cells, whereas STAT5A knockdown has inhibitory effects. Further nude mouse experiments showed that STAT5A knockdown effectively inhibits the growth and metastasis of gastric cancer in vivo. Moreover, as a transcription factor, STAT5A represses KLF4 transcription by binding to its promoter region. The overexpression of KLF4 can counteract the oncogenic impact of STAT5A. Overall, this study highlights the crucial role of m6A in gastric cancer and provides potential therapeutic targets for gastric cancer.

Tailored Triggering of High-Quality Multi-Dimensional Coupled Topological States in Valley Photonic Crystals.

The combination of higher-order topological insulators and valley photonic crystals has recently aroused extensive attentions due to the great potential in flexible and efficient optical field manipulations. Here, we computationally propose a photonic device for the 1550 nm communication band, in which the topologically protected electromagnetic modes with high quality can be selectively triggered and modulated on demand. Through introducing two valley photonic crystal units without any structural alteration, we successfully achieve multi-dimensional coupled topological states thanks to the diverse electromagnetic characteristics of two valley edge states. According to the simulations, the constructed topological photonic devices can realize Fano lines on the spectrum and show high-quality localized modes by tuning the coupling strength between the zero-dimensional valley corner states and the one-dimensional valley edge states. Furthermore, we extend the valley-locked properties of edge states to higher-order valley topological insulators, where the selected corner states can be directionally excited by chiral source. More interestingly, we find that the modulation of multi-dimensional coupled photonic topological states with pseudospin dependence become more efficient compared with those uncoupled modes. This work presents a valuable approach for multi-dimensional optical field manipulation, which may support potential applications in on-chip integrated nanophotonic devices.

Achieving High-Rate and Stable Sodium-Ion Storage by Constructing Okra-Like NiS2/FeS2@Multichannel Carbon Nanofibers.

Transition metal sulfides (TMSs) are considered as promising anode materials for sodium-ion batteries (SIBs) due to their high theoretical capacities. However, the relatively low electrical conductivity, large volume variation, and easy aggregation/pulverization of active materials seriously hinder their practical application. Herein, okra-like NiS2/FeS2 particles encapsulated in multichannel N-doped carbon nanofibers (NiS2/FeS2@MCNFs) are fabricated by a coprecipitation, electrospinning, and carbonization/sulfurization strategy. The combined advantages arising from the hollow multichannel structure in carbon skeleton and heterogeneous NiS2/FeS2 particles with rich interfaces can provide facile ion/electron transfer paths, ensure boosted reaction kinetics, and help maintain the structural integrity, thereby resulting in a high reversible capacity (457 mA h g-1 at 1 A g-1), excellent rate performance (350 mA h g-1 at 5 A g-1), and outstanding long-term cycling stability (93.5% retention after 1100 cycles). This work provides a facile and efficient synthetic strategy to develop TMS-based heterostructured anode materials with high-rate and stable sodium storage properties.

Genomics insights into flowering and floral pattern formation: regional duplication and seasonal pattern of gene expression in Camellia.

BACKGROUND: The formation and domestication of ornamental traits are influenced by various aspects, such as the recognition of esthetic values and cultural traditions. Camellia japonica is widely appreciated and domesticated around the world mainly due to its rich variations in ornamental traits. Ornamental camellias have a diverse range of resources, including different bud variations from Camellia spp. as well as inter- and intra- specific hybridization. Despite research on the formation of ornamental traits, a basic understanding of their genetics and genomics is still lacking. RESULTS: Here, we report the chromosomal-level reference genome of C. japonica through combining multiple DNA-sequencing technologies and obtain a high-density genetic linkage map of 4255 markers by sequencing 98 interspecific F1 hybrids between C. japonica and C. chekiangoleosa. We identify two whole-genome duplication events in C. japonica: one is a shared ancient γ event, and the other is revealed to be specific to genus Camellia. Based on the micro-collinearity analysis, we find large-scale segmental duplication of chromosome 8, resulting to two copies of the AGAMOUS loci, which may play a key role in the domestication of floral shapes. To explore the regulatory mechanisms of seasonal flowering, we have analyzed year-round gene expression patterns of C. japonica and C. azalea-a sister plant of continuous flowering that has been widely used for cross breeding. Through comparative analyses of gene co-expression networks and annual gene expression patterns, we show that annual expression rhythms of some important regulators of seasonal growth and development, including GIGANTEA and CONSTANS of the photoperiod pathway, have been disrupted in C. azalea. Furthermore, we reveal that the distinctive expression patterns of FLOWERING LOCUS T can be correlated with the seasonal activities of flowering and flushing. We demonstrate that the regulatory module involved in GIGANTEA, CONSTANS, and FLOWERING LOCUS T is central to achieve seasonality. CONCLUSIONS: Through the genomic and comparative genomics characterizations of ornamental Camellia spp., we propose that duplication of chromosomal segments as well as the establishment of gene expression patterns has played a key role in the formation of ornamental traits (e.g., flower shape, flowering time). This work provides a valuable genomic platform for understanding the molecular basis of ornamental traits.

Impact of different genetic mutations on granulocyte development and G-CSF responsiveness in congenital neutropenia.

ABSTRACT: Congenital neutropenia (CN) is a genetic disorder characterized by persistent or intermittent low peripheral neutrophil counts, thus increasing susceptibility to bacterial and fungal infections. Various forms of CN, caused by distinct genetic mutations, exhibit differential responses to granulocyte colony-stimulating factor (G-CSF) therapy, with the underlying mechanisms not fully understood. This study presents an in-depth comparative analysis of clinical and immunological features in 5 CN patient groups (severe congenital neutropenia [SCN]1, SCN3, cyclic neutropenia [CyN], warts, hypogammaglobulinaemia, infections and myelokathexis [WHIM], and Shwachman-Bodian-Diamond Syndrome [SBDS]) associated with mutations in ELANE, HAX1, CXCR4, and SBDS genes. Our analysis led to the identification of 11 novel mutations in ELANE and 1 each in HAX1, CXCR4, and G6PC3 genes. Investigating bone marrow (BM) granulopoiesis and blood absolute neutrophil count after G-CSF treatment, we found that SCN1 and SCN3 presented with severe early-stage disruption between the promyelocyte and myelocyte, leading to a poor response to G-CSF. In contrast, CyN, affected at the late polymorphonuclear stage of neutrophil development, showed a strong G-CSF response. WHIM, displaying normal neutrophil development, responded robustly to G-CSF, whereas SBDS, with moderate disruption from the early myeloblast stage, exhibited a moderate response. Notably, SCN1 uniquely impeded neutrophil development, whereas SCN3, CyN, WHIM, and SBDS also affected eosinophils and basophils. In addition, SCN1, SCN3, and CyN presented with elevated serum immunoglobulins, increased BM plasma cells, and higher A Proliferation-Inducing Ligand levels. Our study reveals a strong correlation between the stage and severity of granulocyte development disruption and the efficacy of G-CSF therapy.

Origin differentiation based on volatile constituents of genuine medicinal materials Quisqualis indica L. via HS-GC-MS, response surface methodology, and chemometrics.

INTRODUCTION: Quisqualis indica L. (QIL) has a long history as a traditional Chinese herb in China, but the study of volatile components in QIL from different geographical sources has been relatively rare. OBJECTIVES: To establish an optimal headspace gas chromatography-mass spectrometry (HS-GC-MS) method to comprehensively analyse the volatile component profile and screen quality markers of QIL from different origins. METHODS: Response surface methodology (RSM) was used to optimise the conditions for headspace analysis. The volatile components of QIL from four main origins of southwest China were analysed and identified by HS-GC-MS. The similarity of all samples of QIL was evaluated by fingerprint. The differences of the volatile components in QIL from different origins were distinguished by chemometrics. RESULTS: According to the optimal conditions of RSM, a total of 31 volatile components were identified, including fatty acids, aldehydes, alcohols, alkyl pyrazines, and other volatile components. Similarity evaluation presented that there were 26 common volatile components with different contents in all samples. Principal component analysis (PCA) showed that QIL from four different origins could be roughly divided into four categories. Hierarchical cluster analysis (HCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) indicated that QIL from different origins had obvious regional characteristics. CONCLUSION: The optimised HS-GC-MS method provided a strategy to rapidly, effectively, and accurately elucidate the volatile component profile of QIL from different origins, and seven important differential components were screened for quality evaluation and origin traceability.

Identification of Different Varieties of Oil Peony Seeds Combining ICP-MS with Chemometrics and Assessment of Associated Health Risk.

Peony seed is an excellent oil crop, and peony seed oil is rich in unsaturated fatty acids needed by the human body. In this study, inductively coupled plasma mass spectrometry (ICP-MS), fingerprint, and chemometrics, the correlation between the content of inorganic elements in oil peony seeds, their origins, and varieties were investigated. Meanwhile, estimated daily intake (EDI), target hazard quotient (THQ), hazard index (HI), and carcinogenic risks (CR) were combined to evaluate the comprehensive health risks of heavy metals in peony seed oil. The results showed that the difference in the content of inorganic elements could identify the varieties of oil peony seeds. Sr, K, Ca, V, Al, Fe, Cu, Ba, As, Ga, Co, and Rb were the characteristic inorganic elements that played a role in identification. In addition, The THQs and HIs (< 1) for non-carcinogenic elements indicated no risk. The CRs indicated that the carcinogenic harm was negligible. The study concluded that three varieties of peony seed oil would not pose any health hazard. It provided an effective comprehensive method for the identification of oil peony seeds and predicted the potential health risks of edible peony seed oil, providing a reference for the development and consumption of peony seed oil food.

Identifying cis-Acting Elements Associated with the High Activity and Endosperm Specificity of the Promoters of Genes Encoding Low-Molecular-Weight Glutenin Subunits in Common Wheat (Triticum aestivum).

Low-molecular-weight glutenin subunits (LMW-GSs) associated with bread-baking quality and flour nutrient quality accumulate in endosperms of common wheat and related species. However, the mechanism underlying the expression regulation of genes encoding LMW-GSs has not been fully elucidated. In this study, we identified LMW-D2 and LMW-D7, which are highly and weakly expressed, respectively, via the analysis of RNA-sequencing data of Chinese Spring wheat and wheat transgenic lines transformed with 5' deletion promoter fragments and GUS fusion constructs. The 605-bp fragment upstream of the LMW-D2 start codon could drive high levels of GUS expression in the endosperm. The truncated endosperm box located at the -300 site resulted in the loss of LMW-D2 promoter activity, and a single-nucleotide polymorphism on the GCN4 motif was closely related to the expression of LMW-GSs. TCT and TGACG motifs, as well as the others located on the 5' distal end, might also be involved in the transcription regulation of LMW-GSs. In transgenic lines, fusion proteins of LMW-GS and GUS were deposited into protein bodies. Our findings provide new insights into the mechanism underlying the transcription regulation of LMW-GSs and will contribute to the development of wheat endosperm as a bioreactor for the production of nutraceuticals, antibodies, vaccines, and medicinal proteins.

Mitochondria of Live Mizuhopecten yessoensis Scallops Can Sensitively Respond to Quality Changes during Dry/Reimmersed Storage as Determined by TMT-Labeled Proteomic Analysis.

Dry/reimmersed storage is often used in the transportation of live scallops. In this study, tandem mass tag (TMT)-labeled protein omics were used to quantitatively analyze the protein changes in scallops during dry/reimmersed stress. The results showed that during dry storage, scallops maintained cellular redox homeostasis through the upregulation of SCO1-like protein and thioredoxin domain-containing protein and reduced organic acids from the ATP synthetic process by the downregulation of NADH dehydrogenase, thereby reducing the damage caused during dry storage. During reimmersed storage, mitochondrial proteins underwent very sensitive changes. By upregulating aerobic respiration-related proteins (including proteins involved in glucose phosphate metabolism, glyceraldehyde 3-phosphate metabolism, etc.), the ATP synthesis ability was improved. However, the damage to the mitochondrial structure by dry storage could not be completely recovered, even by reimmersion. This included some apoptosis-related proteins that were obviously upregulated. In summary, compared with ATP-related indexes, mitochondria can respond more sensitively to dry storage stress.

B cell abnormalities and autoantibody production in patients with partial RAG deficiency.

Mutations in the recombination activating gene 1 (RAG1) and RAG2 in humans are associated with a broad spectrum of clinical phenotypes, from severe combined immunodeficiency to immune dysregulation. Partial (hypomorphic) RAG deficiency (pRD) in particular, frequently leads to hyperinflammation and autoimmunity, with several underlying intrinsic and extrinsic mechanisms causing a break in tolerance centrally and peripherally during T and B cell development. However, the relative contributions of these processes to immune dysregulation remain unclear. In this review, we specifically focus on the recently described tolerance break and B cell abnormalities, as well as consequent molecular and cellular mechanisms of autoantibody production in patients with pRD.

Th2/Th1 cytokine imbalance is associated with higher recurrent intussusception incidence in children: A 10-year retrospective study of 1657 patients.

Recurrent intussusception is one of the common acute abdominal diseases in infants, which seriously affects the physical and mental health of infants, but its risk factors have not been fully clarified. The objective of this study was to evaluate the relationship between Th2/Th1 cytokine imbalance and recurrent intussusception, so as to provide a theoretical basis for making a more comprehensive treatment strategy for patients with recurrent intussusception. A retrospective study was conducted between July 2012 and September 2022, enrolling patients with intussusception in Children's Hospital of Hebei Province. The patients were divided into recurrent group and non-recurrent group according to whether they suffered from recurrent intussusception. We summarized the clinical characteristics of recurrent intussusception and explored the role of T helper 2 (Th2)/T helper 1 (Th1) cytokine imbalance in it. A total of 2008 patients were initially enrolled and finally 1657 patients qualified for the study. The results showed that the incidence of recurrent intussusception was 18.41% and the Th2/Th1 cytokine imbalance was closely related to the incidence of recurrent intussusception. Th2/Th1 cytokine imbalance is a potential risk factor of recurrent intussusception and more likely to occur in children between the ages of 2 and 3. Future studies are needed to preemptively target the Th2/Th1 cytokine imbalance to formulate a reasonable treatment plan for children with intussusception to avoid recurrence.

Pericarp starch metabolism is associated with caryopsis development and endosperm starch accumulation in common wheat.

The wheat pericarp is the main component of the caryopsis at the early development stage and ultimately converts into a tissue that covers the mature caryopsis. A large number of starch granules are accumulated in the pericarp, but the production of and the role of starch granules in caryopsis development remain- elusive. In the present study, the relationship between accumulated starch granules and starch metabolism-related genes in wheat pericarp was investigated using paraffin section observations, expression analysis, and mutant analysis. Starch synthesis is initiated before anthesis and is dependent on a sucrose uptake and conversion system similar to that in the endosperm. TaPTST2 is required to initiate the production of pericarp starch granules. Pericarp starch granules gradually disappeared at the filling stage with high expression levels of genes encoding ß-amylase, sucrose-phosphate synthase, and sucrose-phosphate phosphatase. As a maternal tissue adjacent to the endosperm and embryo, the pericarp plays a temporary reservoir for excess nutrients delivered into the caryopsis during the early development stage and exported at the filling stage. The pericarp contributes to the development of the endosperm and embryo as well as the accumulation of endosperm starch. The metabolism of pericarp starch may affect the weight of the wheat caryopsis.

Sourness impacts envy and jealousy in Chinese culture.

In the present study, five experiments (N = 233) were designed to explore whether sourness as a sensory experience could implicitly impact social emotions of envy and jealousy in Chinese culture. Experiment 1 (n = 63) explored the implicit conceptual association between sourness words (vs. bitterness words) and envy/jealousy words. Experiment 2 (n = 70) and 3 (n = 20) examined the priming effects of imagined and tasted sourness (vs. bitterness and sweetness) on self-rated emotional intensity in envy- and jealousy-arousing situations, respectively. Experiment 4 (n = 40) and 5 (n = 40) further testified the priming effects of imagined and tasted sourness (vs. bitterness and sweetness) on self-rated emotional intensity in four types of social situations (i.e., envy, jealousy, sad and happy events), respectively. In the results, sourness was found as the only taste that not only conceptually associated with envy/jealousy, but also significantly primed envy/jealousy feelings. The possible mechanism underlying the association of sourness-envy/jealousy was discussed.

Copper inhibits postacidification of yogurt and affects its flavor: A study based on the Cop operon.

Yogurt and its related products are popular worldwide. During transportation and storage, Lactobacillus delbrueckii ssp. bulgaricus in yogurt continues to metabolize to form lactic acid, the postacidification phenomenon of yogurt. Postacidification of yogurt is a widespread phenomenon in the dairy industry. Many scholars have done research on controlling the postacidification process, but few report on the molecular mechanisms involved. In this study, we used a molecular-assisted approach to screen food additives that can inhibit postacidification and analyzed its effects on yogurt quality as well as its regulatory mechanism from multi-omics perspectives in combination. The copper ion was found to upregulate the expression of the LDB_RS05285 gene, and the copper transporter-related genes were regulated by copper. Based on the metabolic-level analysis, copper was found to promote lactose hydrolysis, accumulate a large amount of glucose and galactose, inhibit the conversion of glucose to lactic acid, and reduce the production of lactic acid. The significantly greater abundance of l-isoleucine and l-phenylalanine increased the abundance of 3-methylbutyraldehyde (∼1.2 times) and benzaldehyde (∼7.9 times) to different degrees, which contributed to the formation of the overall flavor of yogurt. Copper not only stabilizes the acidity of yogurt, but also it improves the flavor of yogurt. Through this established method involving quantitative and correlation analyses at the transcriptional and metabolic levels, this study provides guidance for the research and development of food additives that inhibit postacidification of yogurt and provide a reference for studying the changes of metabolites during storage of yogurt.

Protective role of Cecropin AD against LPS-induced intestinal mucosal injury in chickens.

Introduction: Cecropin AD (CAD), a renowned antimicrobial peptide, has shown promising potential in treating various bacterial infections. This study investigates the protective effects of CAD against lipopolysaccharide (LPS)-induced intestinal adversities in chickens. Methods: Sixty SPF-grade chicks were divided into groups and exposed to different dosages of CAD, followed by LPS administration. The study assessed the impact of CAD on intestinal mucosal injury markers, oxidative stress, and inflammation. Results: LPS significantly increased Diamine oxidase (DAO) and D-lactate (D-LA) levels, both indicators of intestinal mucosal injury. CAD treatment substantially attenuated these elevations, particularly at higher dosages. Additionally, CAD markedly reduced oxidative stress in intestinal tissues, as shown by normalized antioxidant levels and decreased reactive oxygen species. Histological analysis supported these findings, showing better-preserved villi structures in CAD-treated groups. Furthermore, CAD significantly reduced IL-6 and IL-8 expression post-LPS stimulation and effectively regulated the NLRP3 inflammasome pathway, decreasing associated factors like NLRP3, Caspase-1, IL-1b, and IL-18. Discussion: The study demonstrates CAD's therapeutic potential in alleviating LPS-induced intestinal injuries. The protective effects are primarily attributed to its anti-inflammatory and antioxidative actions and modulation of the NLRP3 inflammasome pathway.

Whole-Genome Sequencing-Based Species Classification, Multilocus Sequence Typing, and Antimicrobial Resistance Mechanism Analysis of the Enterobacter cloacae Complex in Southern China.

Members of the Enterobacter cloacae complex (ECC) are important opportunistic nosocomial pathogens that are associated with a great variety of infections. Due to limited data on the genome-based classification of species and investigation of resistance mechanisms, in this work, we collected 172 clinical ECC isolates between 2019 and 2020 from three hospitals in Zhejiang, China and performed a retrospective whole-genome sequencing to analyze their population structure and drug resistance mechanisms. Of the 172 ECC isolates, 160 belonged to 9 classified species, and 12 belonged to unclassified species based on ANI analysis. Most isolates belonged to E. hormaechei (45.14%) followed by E. kobei (13.71%), which contained 126 STs, including 62 novel STs, as determined by multilocus sequence typing (MLST) analysis. Pan-genome analysis of the two ECC species showed that they have an "open" tendency, which indicated that their Pan-genome increased considerably with the addition of new genomes. A total of 80 resistance genes associated with 11 antimicrobial agent categories were identified in the genomes of all the isolates. The most prevailing resistance genes (12/29, 41.38%) were related to ß-lactams followed by aminoglycosides. A total of 247 ß-lactamase genes were identified, of which the blaACT genes were the most dominant (145/247, 58.70%), followed by the blaTEM genes (21/247, 8.50%). The inherent ACT type ß-lactamase genes differed among different species. blaACT-2 and blaACT-3 were only present in E. asburiae, while blaACT-9, blaACT-12, and blaACT-6 exclusively appeared in E. kobei, E. ludwigii, and E. mori. Among the six carbapenemase-encoding genes (blaNDM-1, blaNDM-5, blaIMP-1, blaIMP-4, blaIMP-26, and blaKPC-2) identified, two (blaNDM-1 and blaIMP-1) were identified in an ST78 E. hormaechei isolate. Comparative genomic analysis of the carbapenemase gene-related sequences was performed, and the corresponding genetic structure of these resistance genes was analyzed. Genome-wide molecular characterization of the ECC population and resistance mechanism would offer valuable insights into the effective management of ECC infection in clinical settings. IMPORTANCE The presence and emergence of multiple species/subspecies of ECC have led to diversity and complications at the taxonomic level, which impedes our further understanding of the epidemiology and clinical significance of species/subspecies of ECC. Accurate identification of ECC species is extremely important. Also, it is of great importance to study the carbapenem-resistant genes in ECC and to further understand the mechanism of horizontal transfer of the resistance genes by analyzing the surrounding environment around the genes. The occurrence of ECC carrying two MBL genes also indicates that the selection pressure of bacteria is further increased, suggesting that we need to pay special attention to the emergence of such bacteria in the clinic.

Protocol for an open-label, single-arm, multicentre clinical study to evaluate the efficacy and safety of rituximab in the first episode of paediatric idiopathic nephrotic syndrome.

INTRODUCTION: Rituximab (RTX) effectively prevents relapses in patients with complicated steroid-sensitive nephrotic syndrome (SSNS). The 1-year relapse-free survival rate is approximately 30% in children after the first episode of SSNS treated with standardised corticosteroids. Whether the benefits of RTX extend to the first relapse are unknown. The efficacy and safety of RTX in the first episode of paediatric idiopathic nephrotic syndrome (RTXFIRPedINS) trial (NCT04783675) will assess its effect on the risk of subsequent relapse. METHODS AND ANALYSIS: RTXFIRPedINS is an open-label, single-arm, multicentre trial targeting patients aged 1-18 years with a first episode of SSNS. All patients will receive standardised corticosteroid treatment for 12 weeks. A sample size of 44 patients provides 80% power to detect a 20% increase in the 1-year relapse-free rate, assuming a dropout rate of 10%. After obtaining informed consent and screening, eligible patients will be treated with a single intravenous infusion of 375 mg/m2 RTX within 1 week after achieving remission. Trimethoprim-sulfamethoxazole will be administered for 3 months after RTX administration to prevent Pneumocystis carinii infection. The follow-up period will be 1 year. The primary outcome is the 1-year relapse-free survival rate after RTX infusion. The secondary study outcomes are the number of days from the infusion of RTX to the occurrence of the first relapse, 6-month relapse-free survival rate, the B cell recovery time and treatment-related adverse events. Immunological factors will be studied as predictors of response. ETHICS AND DISSEMINATION: This trial was approved by the Ethics Committee of the Children's Hospital of Fudan University and seven local ethics committees. We will publish our study results in peer-reviewed journals and present them at international scientific meetings. TRIAL REGISTRATION NUMBER: NCT04783675.

Hepatotoxicity and the role of the gut-liver axis in dogs after oral administration of zinc oxide nanoparticles.

Zinc oxide nanoparticles (ZnO NPs) have been used in many fields, and people are concerned about its effects on health. The present study reported the changes in liver metabolites and intestinal microbiota induced by overused ZnO NPs in dogs and explored the related mechanisms of liver injury induced by ZnO NPs. The results showed that overused ZnO NPs promote zinc accumulation in the liver and increase liver coefficient and serum liver-related indexes. In addition, the overuse of ZnO NPs increase the reactive oxygen species levels, affecting the hepatocyte antioxidant capacity and mitochondrial function. Results showed that ZnO NPs significantly inhibited the hepatocyte apoptosis via the Cytc pathway and promoted the autophagy via activating the mTOR/ATG5 pathway. Metabolic analysis of liver tissue showed that 81 metabolites changed overall and mainly affected the glycerophospholipid metabolism. ZnO NPs can significantly change the richness and diversity of the intestinal bacteria in dogs, increasing the abundance of Firmicutes and Actinobacteria while reducing the bacterial abundance of Proteobacteria. In conclusion, the results suggest that overexposure to ZnO NPs can lead to the disruption of intestinal microbiome and liver metabolites in dogs, which ultimately leads to liver damage.