Nutrition impacts of non-solid cooking fuel adoption on under-five children in developing countries

This paper examines the nutrition impacts of using non-solid cooking fuel on under-five children in developing countries.We draw on data from more than 1.12 million children in 62 developing countries from the Demographic and Health Surveys(DHS).Results from both fixed effects(FE)and instrumental variable(IV)estimates show that using non-solid cooking fuel significantly improves the nutrition outcomes of under-five children.Compared with their peers from households mainly using solid fuel,children from households mainly using non-solid fuel exhibit a lower probability of experiencing stunting(by 5.9 percentage points)and being underweight(by 1.2 percentage points).Our further investigation provides evidence for several underlying mechanisms,such as improved indoor air quality,induced reduction in children’s respiratory symptoms,benefits on maternal health,and reduction in maternal time spent on fuel collection or cooking.Heterogenous analyses suggest that the nutrition benefits of using non-solid cooking fuel are more prominent among boys,children above three years old,and those from households of lower socioeconomic status,rural areas,and Southeast Asia.

Towards Lessening Learners’Aversive Emotions and Promoting Their Mental Health:Developing and Validating a Measurement of English Speaking Demotivation in the Chinese EFL Context

While a plethora of studies has been conducted to explore demotivation and its impact on mental health in second language(L2)education,scanty research focuses on demotivation in L2 speaking learning.Particularly,little research explores the measures to quantify L2 speaking demotivation.The present two-phase study attempts to develop and validate an English Speaking Demotivation Scale(ESDS).To this end,an independent sample of 207 Chinese tertiary learners of English as a Foreign Language(EFL)participated in the development phase,and another group of 188 Chinese EFL learners was recruited for the validation of the scale.Exploratory Factor Analysis(EFA)and Confirmatory Factor Analysis(CFA)were employed to determine the factor structure of the scale.The EFA results revealed a six-factor solution with Teacher-related Factors in Learning Spoken English(TFLSE),Interest and Valence in Learning Spoken English(IVLSE),Self-efficacy in Learning Spoken English(SELSE),Negative Peer Influence in Learning Spoken English(NPILSE),Undesirable Environment for Learning Spoken English(UELSE),and Negative Influence of Assessment and Learning Materials in speaking class(NIALM).In the validation phase,Confirmatory Factor Analysis(CFA)was performed to validate the internal structure of the scale.The CFA results showed that the model fits the data well.Overall,the ESDS is a robust and trustworthy psychometric tool that could be utilized to examine L2 speaking demotivation.Implications for diminishing EFL learners’demotivation,lessening their aversive emotions and promoting their mental health are also discussed.

Challenges to establishing and maintaining kidney transplantation programs in developing countries:What are the coping strategies?

Kidney transplantation(KT)is the optimal form of renal replacement therapy for patients with end-stage renal diseases.However,this health service is not available to all patients,especially in developing countries.The deceased donor KT programs are mostly absent,and the living donor KT centers are scarce.Single-center studies presenting experiences from developing countries usually report a variety of challenges.This review addresses these challenges and the opposing strategies by reviewing the single-center experiences of developing countries.The financial challenges hamper the infrastructural and material availability,coverage of transplant costs,and qualification of medical personnel.The sociocultural challenges influence organ donation,equity of beneficence,and regular follow-up work.Low interests and motives for transplantation may result from high medicolegal responsibilities in KT practice,intense potential psychosocial burdens,complex qualification protocols,and low productivity or compensation for KT practice.Low medical literacy about KT advantages is prevalent among clinicians,patients,and the public.The inefficient organizational and regulatory oversight is translated into inefficient healthcare systems,absent national KT programs and registries,uncoordinated job descriptions and qualification protocols,uncoordinated on-site investigations with regulatory constraints,and the prevalence of commercial KT practices.These challenges resulted in noticeable differences between KT services in developed and developing countries.The coping strategies can be summarized in two main mechanisms:The first mechanism is maximizing the available resources by increasing the rates of living kidney donation,promoting the expertise of medical personnel,reducing material consumption,and supporting the establishment and maintenance of KT programs.The latter warrants the expansion of the public sector and the elimination of non-ethical KT practices.The second mechanism is recruiting external resources,including financial,experience,and training agreements.

基于POA的读写结合教学实践--以外研版高中英语教材Developing Ideas板块为例

读写结合是实施综合性语言学习的重要途径。产出导向法(POA)强调通过语言输入为语言输出搭建支架,有助于落实读写结合教学,实现语言文化知识的学习理解和迁移应用。本文以外研版高中英语教材Developing Ideas板块为例,阐述基于POA进行高中英语读写结合教学的策略:(1)输出驱动--呈现任务、尝试写作、激发动机。(2)输入促成--以读促写、搭建支架、迁移应用。(3)以评为学--师生合作、典型分析、完善修订。在此过程中,写作任务设计、阅读文本的选择等都有所要求。

The molecular mechanism of WRINKLED1 transcription factor regulating oil accumulation in developing seeds of castor bean

The transcription factor WRINKLED1(WRI1),a member of AP2 gene family that contain typical AP2 domains,has been considered as a master regulator regulating oil biosynthesis in oilseeds.However,the regulatory mechanism of RcWRI1 in regulating oil accumulation during seed development has not been clearly addressed.Castor bean(Ricinus communis)is one of the most important non-edible oil crops and its seed oils are rich in hydroxy fatty acids,widely applied in industry.In this study,based on castor bean reference genome,three RcWRIs genes(RcWRI1,RcWRI2 and RcWRI3)were identified and the expressed association of RcWRI1 with oil accumulation were determined.Heterologous transformation of RcWRI1 significantly increased oil content in tobacco leaf,confirming that RcWRI1 activate lipid biosynthesis pathway.Using DNA Affinity Purification sequencing(DAP-seq)technology,we confirmed RcWRI1 binding with Transcription Start Site of genes and identified 7961 WRI1-binding candidate genes.Functionally,these identified genes were mainly involved in diverse metabolism pathways(including lipid biosynthesis).Three cis-elements AW-box([CnTnG](n)7[CG])and AW-boxes like([GnAnC](n)6[GC]/[GnAnC](n)7[G])bound with RcWRI1 were identified.Co-expression network analysis of RcWRI1 further found that RcWRI1 might be widely involved in biosynthesis of storage materials during seed development.In particular,yeast one hybrid experiments found that both AP2 domains within RcWRI1 were required in binding targeted genes.These results not only provide new evidence to understand the regulatory mechanism of RcWRI1 in regulation of oil accumulation during castor bean seed development,but also give candidate gene resource for subsequent genetic improvement toward increasing oil content in oilseed crops.

The barriers of joining in vitro fertilization programs among infertile couples in developing countries:A scoping review

Objective:To determine the barriers of joining in-vitro fertilization(IVF)programs among infertile couples in developing countries.Methods:This study assessed infertile couples and the barriers or associated factors resulting in delayed decision-making of joining IVF program by searching databases PubMed,ScienceDirect,EBSCO,ClinicalKey,and Cochrane Library from inception until December 31,2021.Additional search strategies were snowballing literature search and citation tracking.Results:Eleven articles were included in the scoping review.The cost was the greatest barrier of joining IVF program.Limited access and lack of assisted reproductive technology centers,few qualified infertility trained staff,insufficient government support,low priority in government policy,along with sociocultural factors,such as religion and false beliefs or myths were also majorly considered to be associated obstacles.Conclusions:The main barrier associated with IVF program among infertile couples in developing countries is the high cost of the IVF services.

Environmental Impact Assessment System and Process in Developing Countries

Environmental impact assessment (EIA), a decision-making process for project appraisal and sustainability adopted globally as an administrative process to identify, predict, evaluate, and monitor projects from their feasibility, preconstruction, construction, and operation stages to mitigate the adverse impacts and enhance the beneficial impacts for the protection of the affected environment, The study objective is to explore global EIA systems and processes and find shortcomings and implications for making the best instrument or tool to protect the natural environment from man-made activities over the project cycle. For this, the relevant literature on the EIA system and process was reviewed and evaluated through the application of quantitative and qualitative approaches, including the assessment of legal instruments and the adoption of EIA methodologies in developing countries. EIA, initiated in the 1970s in the US and Australia, expanded to developing countries and was amended in East and Southeast Asia from the 1970s to the 2000s. The evaluation assessed that the South Asian countries follow UNEP and IAIA guidelines, utilizing national laws and expert consultations, with screening criteria and administrative processes based on established environmental legislation. Ad hoc, checklist, matrix, network, overlay, cost-benefit analysis, and predictive or simulation in EIA practice are used to assess the environmental impacts of development activities. Failure to recommend major projects undermines public trust and prevents mitigation measures from being implemented. Most developing countries have followed EIA to fulfill the legal requirement with shadow-off monitoring and follow-up rather than to upset, reduce, or compensate for the project impacts as per size, location, and severity of the project area. The research and guidelines outlined in the IAIA principles and process have synthesized the best EIA practices worldwide. Public participation, impact coverage, scientific mitigation, transparent evidence-based approaches, monitoring, follow-up, legitimate approaches, and future appraisal opportunities are major concerns to be included in best EIA practice.

Developing Projects with Low-code Combined with ChatGPT

This paper aims to explore a simpler and more user-friendly way of generating software based on model-driven development.Previous studies have attempted to generate code from domain models,hoping to reduce coding time by increasing modeling time.However,as code tools become more advanced,it is challenging to improve efficiency because models are abstract while implementations are concrete.This paper proposes a novel approach that integrates ChatGPT as a plug-in into the whole R&D process and combines it with our code generation tool to enhance R&D efficiency.We have developed some demos to demonstrate the effectiveness of our approach.According to our evaluation,our approach can save more than 90%of the work in implementing the code generation tool,leaving only about 10%of the work for code review,code improvement,and unit testing.

Digital Development Rights in Developing Countries:Where the Governance Rules for Cross-Border Data Flows

The digital development rights in developing countries are based on establishing a new international economic order and ensuring equal participation in the digital globalization process to achieve people's well-rounded development in the digital society.The relationship between cross-border data flows and the realization of digital development rights in developing countries is quite complex.Currently,developing countries seek to safeguard their existing digital interests through unilateral regulation to protect data sovereignty and multilateral regulation for cross-border data cooperation.However,developing countries still have to face internal conflicts between national digital development rights and individual and corporate digital development rights during the process of realizing digital development rights.They also encounter external contradictions such as developed countries interfering with developing countries'data sovereignty,developed countries squeezing the policy space of developing countries through dominant rules,and developing countries having conflicts between domestic and international rules.This article argues that balancing openness and security on digital trade platforms is the optimal solution for developing countries to realize their digital development rights.The establishment of WTO digital trade rules should inherently reflect the fundamental demands of developing countries in cross-border data flows.At the same time,given China's dual role as a digital powerhouse and a developing country,it should actively promote the realization of digital development rights in developing countries.

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.

Selection and Validation of Reference Genes for Normalization of RT-qPCR Analysis in Developing or Abiotic-Stressed Tissues of Loquat (Eriobotrya japonica)

Loquat(Eriobotrya japonica Lindl.)is a subtropical evergreen fruit tree that produces fruits with abundant nutrients and medicinal components.Confirming suitable reference genes for a set of loquat samples before qRT-PCR experiments is essential for the accurate quantification of gene expression.In this study,eight candidate reference genes were selected from our previously published RNA-seq data,and primers for each candidate reference gene were designed and evaluated.The Cq values of the candidate reference genes were calculated by RT-qPCR in 31 different loquat samples,including 12 subgroups of developing or abiotic-stressed tissues.Different combinations of stable reference genes were screened according to a comprehensive rank,which was synthesized from the results of four algorithms,including the geNorm,NormFinder,BestKeeper andΔCt methods.The screened reference genes were verified by normalizing EjLGA1 in each subgroup.The obtained suitable combinations of reference genes for accurate normalization were GAPDH,EF1αand ACT for floral development;GAPDH,UBCE and ACT for fruit setting;EF1α,GAPDH and eIF2B for fruit ripening;ACT,EF1αand UBCE for leaves under heat stress;eIF2B,UBCE and EF1αfor leaves under freezing stress;EF1α,TUA and UBCE for leaves under salt stress;ACT,EF1αand eIF2B for immature pulp under freezing stress;ACT,UBCE and eIF2B for immature seeds under freezing stress;EF1α,eIF2B and UBCE for both immature pulp and seeds under freezing stress;UBCE,TUB and TUA for red-fleshed fruits under cold-storage stress;eIF2B,RPS3 and TUB for white-fleshed fruits under coldstorage stress;and eIF2B,UBCE and RPS3 for both red-and white-fleshed fruits under cold-storage stress.This study obtained different combinations of stable reference genes for accurate normalization in twelve subgroups of developing or abiotic-stressed tissues in loquat.To our knowledge,this is the first report to obtain stable reference genes for normalizing gene expression of abiotic-stressed tissues in E.japonica.The use of the three most stable reference genes could increase the reliability of future quantification experiments.

Linking Competitors’ Knowledge and Developing Innovative Products Using Data Mining Techniques

In this article, the relationship between the knowledge of competitors and the development of new products in the field of capital medical equipment has been investigated. In order to identify the criteria for measuring competitors’ knowledge and developing new capital medical equipment products, marketing experts were interviewed and then a researcher-made questionnaire was compiled and distributed among the statistical sample of the research. Also, in order to achieve the goals of the research, a questionnaire among 100 members of the statistical community was selected, distributed and collected. To analyze the gathered data, the structural equation modeling (SEM) method was used in the SMART PLS 2 software to estimate the model and then the K-MEAN approach was used to cluster the capital medical equipment market based on the knowledge of actual and potential competitors. The results have shown that the knowledge of potential and actual competitors has a positive and significant effect on the development of new products in the capital medical equipment market. From the point of view of the knowledge of actual competitors, the market of “MRI”, “Ultrasound” and “SPECT” is grouped in the low knowledge cluster;“Pet MRI”, “CT Scan”, “Mammography”, “Radiography, Fluoroscopy and CRM”, “Pet CT”, “SPECT CT” and “Gamma Camera” markets are clustered in the medium knowledge. Finally, “Angiography” and “CBCT” markets are located in the knowledge cluster. From the perspective of knowledge of potential competitors, the market of “angiography”, “mammography”, “SPECT” and “SPECT CT” in the low knowledge cluster, “CT scan”, “radiography, fluoroscopy and CRM”, “pet CT”, “CBCT” markets in the medium knowledge cluster and “MRI”, “pet MRI”, “ultrasound” and “gamma camera” markets in the high knowledge cluster are located.

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).

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 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.

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.

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.

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.

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%.

A cell transcriptomic profile p ovides insights into adipocytes of porcine mammary gland across development

Background Studying the composition and developmental mechanisms in mammary gland is crucial for healthy growth of newborns. The mammary gland is inherently heterogeneous, and its physiological function dependents on the gene expression of multiple cell types. Most studies focused on epithelial cells, disregarding the role of neighboring adipocytes.Results Here, we constructed the largest transcriptomic dataset of porcine mammary gland cells thus far. The dataset captured 126,829 high-quality nuclei from physiological mammary glands across five developmental stages(d 90 of gestation, G90;d 0 after lactation, L0;d 20 after lactation, L20;2 d post natural involution, PI2;7 d post natural involution, PI7). Seven cell types were identified, including epithelial cells, adipocytes, endothelial cells, fibroblasts cells, immune cells, myoepithelial cells and precursor cells. Our data indicate that mammary glands at different developmental stages have distinct phenotypic and transcriptional signatures. During late gestation(G90), the differentiation and proliferation of adipocytes were inhibited. Meanwhile, partly epithelial cells were completely differentiated. Pseudo-time analysis showed that epithelial cells undergo three stages to achieve lactation, including cellular differentiation, hormone sensing, and metabolic activation. During lactation(L0 and L20), adipocytes area accounts for less than 0.5% of mammary glands. To maintain their own survival, the adipocyte exhibited a poorly differentiated state and a proliferative capacity. Epithelial cells initiate lactation upon hormonal stimulation. After fulfilling lactation mission, their undergo physiological death under high intensity lactation. Interestingly, the physiological dead cells seem to be actively cleared by immune cells via CCL21-ACKR4 pathway. This biological process may be an important mechanism for maintaining homeostasis of the mammary gland. During natural involution(PI2 and PI7), epithelial cell populations dedifferentiate into mesenchymal stem cells to maintain the lactation potential of mammary glands for the next lactation cycle.Conclusion The molecular mechanisms of dedifferentiation, proliferation and redifferentiation of adipocytes and epithelial cells were revealed from late pregnancy to natural involution. This cell transcriptomic profile constitutes an essential reference for future studies in the development and remodeling of the mammary gland at different stages.