Exceptionally Preserved Punctatus triangulicostalis and Its Post-embryonic Development

Objective The putative coronatean Punctatus flourished at the beginning of the Cambrian explosion, and is significant in the exploration of the origin and evolution of early life. Liu Yunhuan established a new species： Punctatus triangulicostalis Liu, 2006. Two questions remain： how was the special body plan of P. triangulicostalis formed？ How does P. triangulicostalis transform from embryo to the long cylindrical mature individual？ Although the study of Steiner et al. in 2004 involved the above questions, they did not explain them clearly due to the lack of fossil materials at that time. This article aims to consider the above questions based on new information from several rare specimens.

The Post-Embryonic Development of Digestive System and the Demand of Energy of Hucho taimen

With the technology of morphology, histology and nutrition, the characteristics of organ development and the source and demand of nutritional at different stages in post-embryonic （water temperature 3-14℃） of Hucho taimen were discussed. The results showed that the membrane of H. taimen fertilized ovum was ruptured in 30 d, the prototype of U-type stomach was formed, the mouth could open and close freely, and nutrition was supplied from its yolk sac. The membrane ruptured after 8 d, teeth formed, and anus communicated with outsiders. The oropharyngeal cavity, esophagus, U-type stomach, intestine, and anus initially formed in the digestive tube, which nutrition was supplied from the huge yolk sac. The membrane ruptured after 16 d, pancreas and pyloric caeca appeared, larvae could intake and digest exogenous food, fish mixed nutrition, which was from yolk sac and outsiders, mainly depended on endogenous. The membrane ruptured after 24 d, the fish began floating and intaking actively, the fish mixed nutrition which mainly depended on exogenous. The membrane ruptured after 30 d, the yolk sac was completely absorbed, the function and structure of each digestive organs developed consummately, fish was from mixed nutrition into the stage of exogenous nutrition, the nutritional need of juveniles was below 10% fat, level of the optimal protein was 50%.

GAMYB transcription factor LoMYB65 from lily plays a vital role in pollen development

Lily(Lilium spp.)is an important horticultural crop,but its use is limited due to serious pollen contamination problems.There are many studies on pollen development in model plants,but few on flower crops such as lilies.Gibberellin(GA)is a large class of hormones and plays an important role in plant vegetative growth and reproductive development.GAMYB is a group of the R2R3-MYB family upregulated by gibberellin,and plays an important role in anther development.Here,we isolated a novel GAMYB,named LoMYB65,from lily,which was closely related to the AtMYB65 and AtMYB33 in Arabidopsis.Fluorescence quantitative PCR results showed that LoMYB65 was mainly expressed in lily anthers.LoMYB65 could be activated by 288μmol·L^(-1)GA3treatment and the LoMYB65 protein was located in the nucleus and cytoplasm,and had transactivation in yeast and tobacco leaf cells.The conserved motif within 226 amino acids of the C-terminal of LoMYB65 contributed to its transactivation.Overexpression of LoMYB65 caused dwarf phenotype,unnormal tapetum development,less seeds of siliques in transgenic Arabidopsis plants,the transgenic plants showed partly male sterile.Simultaneously,silencing of LoMYB65 with VIGS(Virus Induced Gene Silencing)in lily anthers caused unnormal pollen development and reduced the pollen amount.Overexpression of LoMYB65 in Arabidopsis and silencing of LoMYB65 in lily resulted in decreased pollen counts,so we speculate that LoMYB65 may be dose-dependent.Overall,these findings suggest that LoMYB65 may play an important role in anther development and pollen formation in lily.LoMYB65 may provide a useful candidate gene for pollenless breeding of lily.

Conventional Geothermal Systems and Unconventional Geothermal Developments: An Overview

This paper provides an overview of conventional geothermal systems and unconventional geothermal developments as a common reference is needed for discussions between energy professionals. Conventional geothermal systems have the heat, permeability and fluid, requiring only drilling down to °C, normal heat flow or decaying radiogenic granite as heat sources, and used in district heating. Medium-temperature (MT) 100°C - 190°C, and high-temperature (HT) 190°C - 374°C resources are mostly at plate boundaries, with volcanic intrusive heat source, used mostly for electricity generation. Single well capacities are °C - 500°C) and a range of depths (1 m to 20 Km), but lack permeability or fluid, thus requiring stimulations for heat extraction by conduction. HVAC is 1 - 2 m deep and shallow geothermal down to 500 m in wells, both capturing °C, with °C are either advanced by geothermal developers at <7 Km depth (Enhanced Geothermal Systems (EGS), drilling below brittle-ductile transition zones and under geothermal fields), or by the Oil & Gas industry (Advanced Geothermal Systems, heat recovery from hydrocarbon wells or reservoirs, Superhot Rock Geothermal, and millimeter-wave drilling down to 20 Km). Their primary aim is electricity generation, relying on closed-loops, but EGS uses fractures for heat exchange with earthquake risks during fracking. Unconventional approaches could be everywhere, with shallow geothermal already functional. The deeper and hotter unconventional alternatives are still experimental, overcoming costs and technological challenges to become fully commercial. Meanwhile, the conventional geothermal resources remain the most proven opportunities for investments and development.

Development and technology status of energy storage in depleted gas reservoirs

Utilizing energy storage in depleted oil and gas reservoirs can improve productivity while reducing power costs and is one of the best ways to achieve synergistic development of"Carbon Peak–Carbon Neutral"and"Underground Resource Utiliza-tion".Starting from the development of Compressed Air Energy Storage(CAES)technology,the site selection of CAES in depleted gas and oil reservoirs,the evolution mechanism of reservoir dynamic sealing,and the high-flow CAES and injection technology are summarized.It focuses on analyzing the characteristics,key equipment,reservoir construction,application scenarios and cost analysis of CAES projects,and sorting out the technical key points and existing difficulties.The devel-opment trend of CAES technology is proposed,and the future development path is scrutinized to provide reference for the research of CAES projects in depleted oil and gas reservoirs.

BMPRⅡ^(+)neural precursor cells isolated and characterized from organotypic neurospheres:an in vitro model of human fetal spinal cord development

Roof plate secretion of bone morphogenetic proteins(BMPs)directs the cellular fate of sensory neurons during spinal cord development,including the formation of the ascending sensory columns,though their biology is not well understood.Type-ⅡBMP receptor(BMPRⅡ),the cognate receptor,is expressed by neural precursor cells during embryogenesis;however,an in vitro method of enriching BMPRⅡ^(+)human neural precursor cells(hNPCs)from the fetal spinal cord is absent.Immunofluorescence was undertaken on intact second-trimester human fetal spinal cord using antibodies to BMPRⅡand leukemia inhibitory factor(LIF).Regions of highest BMPRⅡ^(+)immunofluorescence localized to sensory columns.Parenchymal and meningeal-associated BMPRⅡ^(+)vascular cells were identified in both intact fetal spinal cord and cortex by co-positivity with vascular lineage markers,CD34/CD39.LIF immunostaining identified a population of somas concentrated in dorsal and ventral horn interneurons,mirroring the expression of LIF receptor/CD118.A combination of LIF supplementation and high-density culture maintained culture growth beyond 10 passages,while synergistically increasing the proportion of neurospheres with a stratified,cytoarchitecture.These neurospheres were characterized by BMPRⅡ^(+)/MAP2ab^(+/–)/βⅢ-tubulin^(+)/nestin^(–)/vimentin^(–)/GFAP^(–)/NeuN^(–)surface hNPCs surrounding a heterogeneous core ofβⅢ-tubulin^(+)/nestin^(+)/vimentin^(+)/GFAP^(+)/MAP2ab^(–)/NeuN^(–)multipotent precursors.Dissociated cultures from tripotential neurospheres contained neuronal(βⅢ-tubulin^(+)),astrocytic(GFAP+),and oligodendrocytic(O4+)lineage cells.Fluorescence-activated cell sorting-sorted BMPRⅡ^(+)hNPCs were MAP2ab^(+/–)/βⅢ-tubulin^(+)/GFAP^(–)/O4^(–)in culture.This is the first isolation of BMPRⅡ^(+)hNPCs identified and characterized in human fetal spinal cords.Our data show that LIF combines synergistically with high-density reaggregate cultures to support the organotypic reorganization of neurospheres,characterized by surface BMPRⅡ^(+)hNPCs.Our study has provided a new methodology for an in vitro model capable of amplifying human fetal spinal cord cell numbers for>10 passages.Investigations of the role BMPRⅡplays in spinal cord development have primarily relied upon mouse and rat models,with interpolations to human development being derived through inference.Because of significant species differences between murine biology and human,including anatomical dissimilarities in central nervous system(CNS)structure,the findings made in murine models cannot be presumed to apply to human spinal cord development.For these reasons,our human in vitro model offers a novel tool to better understand neurodevelopmental pathways,including BMP signaling,as well as spinal cord injury research and testing drug therapies.

FE65:a hub for neurodevelopment

FE65,initially identified as a binding partner of amyloid precursor protein(APP),is an adaptor protein enriched in the brain and regulated during development.FE65 belongs to the FE65 protein family.This family is comprised of three members,FE65,FE65 like-1(FE65L1),and FE65 like-2(FE65L2).

Constructing a standard system of city sustainable development assessment technology based on the SDGs

Given the rapid development of China’s new urbanization,cities with different locations and varying functional positioning,resource endowments,and development stages have insufficient scientific and applicable technical tools for implementing the United Nations Sustainable Development Goals(SDGs).City managers and policymakers must urgently establish SDG benchmarks to diagnose city development.Moreover,successful experiences from similar cities regarding sustainable development and self-improvement must be learned from to promote diversified,sustainable development across the country.Furthermore,emerging technologies such as artificial intelligence,the Internet of Things,big data and 5G are widely used in smart cities.Therefore,there is a growing need for“knowledge-based,personalized and intelligent”technologies to support monitoring,evaluation,and decision-making processes facilitating sustainable development in cities.This paper uses standardization as the theoretical support and technical basis.This approach can help clarify the sustainable development processes in China and clarify the evaluation results of and provide data on horizontal city comparisons,which can be used to develop evaluation technology for sustainable development in cities and construct a standardized system.The results provide a standard framework for intelligent assessment and decision-making regarding cities’sustainable development capabilities in China.Evaluating major international standardization institutions reveals that the practices of Chinese national standards should be fully absorbed and integrated to guide the evaluation of smart,resilient,and low-carbon cities.To this end,an indicator library of city sustainable development is proposed to provide standard evaluation technology methods.Finally,analyzing the response relationship of the indicator library to SDGs reveals the need for a standardized knowledge map of sustainable development assessment techniques and methods from the perspective of integrated management for sustainable development in cities.

Spatial Differentiation and Convergence Trend of High-quality Development Level of China’s Tourism Economy

This paper aims to interpret the connotation of high-quality development of tourism economy(HQTE)from the perspective of the new development concepts of innovation,coordination,green,openness and sharing,and then to evaluate the spatial differenti-ation of China’s HQTE based on provincial panel data from 2009 to 2018.Specifically,we employ the spatial convergence model to ex-plore the absolute and conditionalβconvergence trends of HQTE in the whole country and the eastern,central and western regions of China.Our empirical results reveal that:1)within the decade,from 2009 to 2018,regions of China with the highest HQTE index is its eastern region followed by the central region and then the western region,but the fastest growing one is the western region of China fol-lowed by the central region and then the eastern region.2)Whether or not the spatial effect is included,there are absolute and condition-alβconvergence in HQTE in the whole country and aforementioned three regions.3)The degree of government attention as well as the level of economic development and location accessibility are the positive driving factors for the convergence of HQTE in the whole country and the three regions.The degree of marketization and human capital have not passed the significance test either in the whole country or in the three regions.The above conclusions could deepen the understanding of the regional imbalance and spatial conver-gence characteristics of HQTE,clarify the primary development objects,and accomplish the goal of China’s HQTE.

Development and Validation of a UPLC Method for the Determination of Docetaxel and Its Related Substances in Pharmaceutical Dosage Forms, an Antineoplastic Agent

A novel, simple, and sensitive Ultra Performance Liquid Chromatography (UPLC) method was developed and validated for the quantification of process-related impurities and degradants, as well as the assay of Docetaxel. The stability-indicating capability of the method was demonstrated through forced degradation studies and a comprehensive mass balance evaluation. Chromatographic separation was achieved using an ACQUITY UPLC BEH C18 column (100 × 2.1 mm, 1.7 µm), with gradient elution. The mobile phase A comprised a mixture of water, methanol, and acetonitrile (500:300:200, v/v/v), while mobile phase B was acetonitrile and water (800:200, v/v). The flow rate was set at 0.4 mL/min, with detection at 232 nm using a photodiode array detector. The method exhibited excellent performance, with a tailing factor of 1.10 for Docetaxel. The method was rigorously validated for precision, accuracy, linearity, LOD, LOQ, ruggedness, specificity, and robustness. Forced degradation studies confirmed the method’s suitability for stability analysis. Stability testing on the drug substance was conducted following ICH guidelines.

The HD-Zip transcription factor LcATHB15 is involved in litchi seed development by promoting the expression of the splicing regulator LcSR45

Litchi (Litchi chinensis Sonn.) is a well-known tropical and subtropical woody fruit that originated from southern China (Wei et al.,2017;Hu et al.,2022).Based on the development of the embryo,litchi varieties can be divided into three distinct groups:large-seeded cultivars,small (abortive)-seeded cultivars and partial abortive-seeded cultivars.Seedless or the reduction in seed size due to embryo abortion is a desirable trait in fruit trees as it directly influences the texture and economic value of fleshy fruits,making small (abortive)-seeded litchi cultivars the most preferred.However,in current production practices,there is no efficient method available to induce or promote high seed abortion in litchi fruit.Therefore,understanding the stenospermy mechanisms of litchi seed development is of great significance for improving agricultural practices and managing genetic resources.

Embryonic and Larval Development of Reciprocal Crosses between Clarias gariepinus (Burchell, 1822) and Clarias jaensis (Boulenger, 1909) in West Cameroon

Establishing intraspecific breeding and hybridization programs and determining genetic variability are two important issues for aquaculture. However, interspecific hybridization to improve growth and feeding efficiency is limited. For this purpose, the embryonic and larval development of reciprocal crosses of Clarias gariepinus (Burchell, 1822) and Clarias jaensis (Boulenger, 1909) were studied under laboratory conditions. The fertilization rate varied from 63.33% to 92%, while the hatching rate ranged from 55.68% to 76% with the highest value in hybrids ♀Cg × ♂Cj. Crosses between ♀Cj × ♂Cj, ♀Cg × ♂Cj and ♀Cj × ♂Cg had embryonic stages similar to those of the pure sib ♀Cg x ♂Cg. All crosses, however, had different timing for the various embryological stages. Hatching occurred at 32 h 15 min and 38 h for ♀Cj × ♂Cj and ♀Cj × ♂Cg, and 23 h and 23 h 30 min, respectively, for ♀Cg × ♂Cg and ♀Cg × ♂Cj. However, both crosses produced viable larvae until the first external feeding. The external morphological features of the larvae were completely formed by the 10th day after hatching. The body forms of the crosses at this time were indistinguishable from the pure sib. This study thus laid the groundwork for further comparative studies on hybrid performance and characterization.

Role of self-help groups on socioeconomic development and the achievement of Sustainable Development Goals(SDGs)among rural women in Cooch Behar District,India

This study examines the transformative role of self-help groups(SHGs)in the socioeconomic development of rural women in Cooch Behar District,India,and their contribution toward achieving Sustainable Development Goals(SDGs)of the United Nations.In this study,we explored the effect of SHGs on rural women by specifically addressing SDGs,such as no poverty(SDG 1),zero hunger(SDG 2),good health and well-being(SDG 3),quality education(SDG 4),and gender equality(SDG 5).Given this issue,a cross-sectional survey and comparison analyses are needed to assess the socioeconomic development of rural women and their awareness level before and after the participation of rural women in SHGs.The survey conducted as part of this study was divided into three sections,namely,demographic characteristics,socioeconomic development,and awareness level,with each focusing on different aspects.A group of 400 individuals who were part of SHGs completed the questionnaire survey form.The results showed that the participation of rural women in SHGs significantly improved their socioeconomic development and awareness level,as supported by both mean values and t test results.Memberships in SHGs and microcredit programs were the major elements that boosted the socioeconomic development of rural women,which also achieves SDGs 1,2,3,4,and 5.This study revealed that participation in SHGs and related financial services significantly aided rural women in economically disadvantaged communities in accumulating savings and initiating entrepreneurial ventures.Moreover,participation in SHGs was instrumental in enhancing the self-confidence,self-efficacy,and overall self-esteem of rural women.Finally,doing so enabled them to move more freely for work and other activities and to make family and common decisions.

Development and formation of wing cuticle based on transcriptomic analysis in Locusta migratoria during metamorphosis

Wings are an important flight organ of insects.Wing development is a complex process controlled by a series of genes.The flightless wing pad transforms into a mature wing with the function of migratory flight during the nymphto-adult metamorphosis.However,the mechanism of wing morphogenesis in locusts is still unclear.This study analyzed the microstructures of the locust wing pads at pre-eclosion and the wings after eclosion and performed the comparative transcriptome analysis.RNA-seq identified 25,334 unigenesand 3,430 differentially expressed genes(DEGs)(1,907 up-regulated and 1,523 down-regulated).The DEGs mainly included cuticle development(LmACPs),chitin metabolism(Lm Idgf4),lipid metabolism-related genes,cell adhesion(Integrin),zinc finger transcription factors(LmSalm,LmZF593 andLmZF521),and others.Functional analysis based on RNA interference and hematoxylin and eosin(H&E)staining showed that the three genes encoded zinc finger transcription factors are essential for forming wing cuticle and maintaining morphology in Locusta migratoria.Finally,the study found that the LmSalm regulates the expression of LmACPs in the wing pads at pre-eclosion,and LmZF593 and LmZF521 regulate the expression of LmIntegrin/LmIdgf4/LmHMT420 in the wings after eclosion.This study revealed that the molecular regulatory axis controls wing morphology in nymphal and adult stages of locusts,offering a theoretical basis for the study of wing development mechanisms in hemimetabolous insects.

OsPPG encodes a pseudouridine-5'-phosphate glycosidase and regulates rice spikelet development

Florets are the basic structural units of spikelets,and their morphogenesis determines the yield and quality of rice grains.However,whether and how pseudouridine-5’-phosphate glycosidase participates in rice spikelet development remains an open question.In this study,we identified a novel gene,OsPPG,which encodes a peroxisome-localized pseudouridine-5’-phosphate glycosidase and regulates the development of rice spikelets.osppg mutants exhibited abnormal sterile lemma,lemma,palea,lodicule,stamens,and pistils;male sterility;shorter panicles;and reduced plant height.OsPPG was found to regulate several OsMADS genes,thereby affecting the morphogenesis of rice spikelets.Furthermore,metabolomics revealed that the OsPPG gene was involved in the decomposition of pseudouridine via the pyrimidine metabolism pathway and may affect the jasmonic acid signaling pathway.These results suggest that OsPPG is a key regulator of rice spikelet development.

Assessment of rural sustainable development and analysis and prediction of obstacles and coupled coordinated development:A case study of Zaozhuang City

In China,the Innovation Demonstration Zone for the National Sustainable Development Agenda is a key initiative for executing the United Nations(UN)2030 Agenda for Sustainable Development.The Zone also plays critical roles in addressing the developmental bottlenecks that China faces and sharing Chinese insights into global sustainable development efforts.On July 15,2022,the State Council endorsed the Zone’s establishment in Zaozhuang City,Shandong Province,where it focuses on innovation-led sustainable development in rural areas.Research into the levels,impediments,interdependencies,and evolutionary trends of rural sustainable development is crucial.Therefore,this research aimed to assist in comprehensively assessing developmental challenges and facilitating the harmonious advancement of social,economic,and environmental aspects in rural areas.In pursuit of the three fundamental dimensions of the UN’s Sustainable Development Goals(SDGs),namely development’s drivers,quality,and equity,this study was grounded in China’s national Rural Revitalization Strategy and the demands of sustainable development strategies.It also aligns with the UN 2030 Agenda for Sustainable Development and the associated SDG indicators.Focusing on four key areas,namely production elements,natural elements,social elements,and rural governance,this study developed an evaluation index system for assessing the level of rural sustainable development.It employed a range of analytical models,including the game theory-based combination empowerment method,barrier degree model,coupling degree model,coupling coordination degree model,and gray prediction GM(1,1)model,to analyze the status and evolving trends of rural sustainable development in Zaozhuang City from 2015 to 2022.The key findings were as follows:①Relative to the baseline year 2015,the sustainable development level in Zaozhuang’s rural areas has shifted toward an improved state overall.②The primary barrier to achieving rural sustainable development in Zaozhuang is the city’s rural governance system.③While the components of rural sustainable development in Zaozhuang are in the early stages of both basic and moderate coordination,an overall enhancement has occurred in their integrative coordination.④Between 2023 and 2025,the level of integrative coordination in Zaozhuang is expected to rise steadily.However,reaching a state of advanced coordination will require additional time for development.

Research on the Development of Smart Animal Husbandry in Beijing

With the rapid development of agricultural science and technology,animal husbandry,as an important pillar in the field of agriculture,is gradually moving towards a new era of smart animal husbandry with the deep integration of informatization and digitalization.This transformation not only breaks through the traditional production mode of animal husbandry,but also promotes it to a new form under the Internet ecology,draws a new blueprint for the development of agriculture and animal husbandry,and gives birth to numerous potential business opportunities for the development of new agriculture.However,the practice and promotion of smart animal husbandry is not a smooth road,and many challenges and problems need to be solved urgently.On the basis of an in-depth investigation of the development status of smart animal husbandry in Beijing,this paper comprehensively analyzes the current problems,including the difficulty of technology integration,the lack of talent reserve,and the need to improve the policy environment.In view of these problems,it puts forward a series of practical suggestions,in order to speed up the development of animal husbandry in Beijing to the direction of smart development,and realize the sustainable development of animal husbandry.

Identification and Expression Profile of a Neuropeptide LFRFamide-Like Gene During Different Stages of Gonadal Development in the Cephalopod Sepia pharaonis

Neuropeptides are widely distributed in vertebrates and invertebrates,regulating a variety of physiological activities in the organisms,such as metabolism,feeding and reproduction.In this study,to explore the function of neuropeptide LFRFamide in Sepia pharaonis,the full-length cDNA of LFRFamide-like gene(named SpLFRFL,MG869822.1)was identified with rapid amplification of cDNA ends(RACE)method.The sequence of SpLFRFL was 860 bp in length and encoded 188 amino acids containing 4 different mature peptides:1 copy of PHTPFRFamide,NSLFRFamide,TIFRFamide,and 3 copies of GNLFRFamide.Multiple alignment and phylogenetic analysis results showed that SpLFRFL shared high identity with LFRFamides of Sepia officinalis and Sepiella japonica and had the closest relationship with them.Through quantitative Real-time PCR(qRT-PCR),it was found that the SpLFRFL gene was highly expressed in the optic lobe and brain at three different stages during gonad development in both genders.Moreover,the four mature peptides at a concentration of 0.01μmol L^(−1) could inhibit the protein synthesis in the Chinese hamster ovary cell strain-K1(CHOK1)induced by SpGnRH.These data suggest that SpLFRFL might be involved in the development and reproduction of S.pharaonis.The results can contribute to future studies on neuropeptide evolution and function and benefit the cuttlefish farming.

Robustness Study and Superior Method Development and Validation for Analytical Assay Method of Atropine Sulfate in Pharmaceutical Ophthalmic Solution

Background: The robustness is a measurement of an analytical chemical method and its ability to contain unaffected by little with deliberate variation of analytical chemical method parameters. The analytical chemical method variation parameters are based on pH variability of buffer solution of mobile phase, organic ratio composition changes, stationary phase (column) manufacture, brand name and lot number variation;flow rate variation and temperature variation of chromatographic system. The analytical chemical method for assay of Atropine Sulfate conducted for robustness evaluation. The typical variation considered for mobile phase organic ratio change, change of pH, change of temperature, change of flow rate, change of column etc. Purpose: The aim of this study is to develop a cost effective, short run time and robust analytical chemical method for the assay quantification of Atropine in Pharmaceutical Ophthalmic Solution. This will help to make analytical decisions quickly for research and development scientists as well as will help with quality control product release for patient consumption. This analytical method will help to meet the market demand through quick quality control test of Atropine Ophthalmic Solution and it is very easy for maintaining (GDP) good documentation practices within the shortest period of time. Method: HPLC method has been selected for developing superior method to Compendial method. Both the compendial HPLC method and developed HPLC method was run into the same HPLC system to prove the superiority of developed method. Sensitivity, precision, reproducibility, accuracy parameters were considered for superiority of method. Mobile phase ratio change, pH of buffer solution, change of stationary phase temperature, change of flow rate and change of column were taken into consideration for robustness study of the developed method. Results: The limit of quantitation (LOQ) of developed method was much low than the compendial method. The % RSD for the six sample assay of developed method was 0.4% where the % RSD of the compendial method was 1.2%. The reproducibility between two analysts was 100.4% for developed method on the contrary the compendial method was 98.4%.

Combining genome-wide association study and transcriptome analysis to identify molecular markers and genetic basis of population-asynchronous ovarian development in Coilia nasus

Coilia nasus,a migratory fish species found in the middle and lower reaches of the Yangtze River and along offshore areas of China,possesses considerable aquacultural and economic potential.However,the species faces challenges due to significant variation in the gonadal development rate among females,resulting in inconsistent ovarian maturation times at the population level,an extended reproductive period,and limitations on fish growth rate due to ovarian prematurity.In the present study,we combined genome-wide association study(GWAS)and comparative transcriptome analysis to investigate the potential single nucleotide polymorphisms(SNPs)and candidate genes associated with population-asynchronous ovarian development in C.nasus.Genotyping of the female population based on whole-genome resequencing yielded 2120695 high-quality SNPs,39 of which were suggestively associated with ovarian development.Of note,a significant SNP peak on LG21 containing 30 suggestively associated SNPs was identified,with cpne5a determined as the causal gene of the peak.Therefore,single-marker and haplotype association analyses were performed on cpne5a,revealing four genetic markers(P<0.05)and seven haplotypes(r2>0.9)significantly associated with the phenotype.Comparative transcriptome analysis of precociously and normally maturing individuals screened out 29 and 426 overlapping differentially expressed genes in the brain and ovary,respectively,between individuals of different body sizes.Integrating the GWAS and transcriptome analysis results,this study identified genes and pathways related to hypothalamic-pituitary-gonadal axis hormone secretion,extracellular matrix,angiogenesis,and gap junctions involved in population-asynchronous ovarian development.The insights gained from this study provide a basis for a deeper understanding of the molecular mechanisms underlying ovarian development in fish and may facilitate the genetic breeding of C.nasus strains exhibiting population-synchronous ovarian development in the future.