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Estimation of plant community composition, aboveground biomass and carbon stock is crucial for understanding forest ecology, strengthening environmental management, and developing effective tools and policies for forest restoration. This study was conducted in nine different forest reserves in Sri Lanka from 2012 to 2018 to examine the impact of community-based forest restoration (CBFR) on stand structural attributes, aboveground biomass, and carbon stock compared to state-managed forests. In total, 180 plots (90 plots in community-managed restoration blocks (CMRBs) and 90 plots assigned to state-managed restoration blocks (SMRBs)) were sampled at the study site. To conduct an inventory of standing trees, circular plots with a radius of 12.6 m (equivalent to an area of 500 square meters) were established. The Shannon diversity index, Allometric equations and Difference in Differences (DID) estimation were used to assess the data. Our study provides evidence of the positive impact of the CBFR program on enriching trees diversity. Considering stand structural attributes of both blocks showed higher trees density in the smaller diameter at breast height (DBH) category, indicating growth in both CMRBs and SMRBs. The results showed that tree biomass and carbon density were disproportionally distributed across the nine different forest reserves. On average, tree biomass and carbon density were higher in SMRBs (79.97 Mg ha−1, 37.58 Mg C ha−1) compared to CMRBs (33.51 Mg ha−1, 15.74 Mg C ha−1). However, CMRBs in Madigala reserve represent the highest biomass (56.53 and 59.92 Mg ha−1) and carbon density (26.57 and 28.16 Mg C ha−1). The results of biomass and carbon estimates were higher in all SMRBs in the nine different forest reserves compared to CMRBs. The findings suggest that future forest restoration programs in Sri Lanka should enhance participatory approaches to optimize tree species diversity, density and carbon storage, particularly in community-controlled forests. Our findings could assist developing tropical nations in understanding how CBFR impacts forest restoration objectives and improves the provision of ecological services within forests.

Abstract The return of rural migrant workers to employment is an endogenous force that drives the development of the rural economy and plays a vital role in rural revitalization. Using data from the China Migrants Dynamic Survey (CMDS) conducted in 2017, this study comprehensively examines the influence of social exclusion and the role of identity on migrant workers’ willingness to return home (MWRH). The findings indicate a significant impact of social exclusion on MWRH, and this relationship remains robust even after employing instrumental variables to address endogeneity concerns. Identity emerges as a crucial mediating factor through which social exclusion affects MWRH, demonstrating a partial mediating effect. Notably, the impact of social exclusion on MWRH is particularly prevalent among rural residents in the western and northeastern regions, the older generation, those with lower incomes, and those with limited intra-provincial mobility. Promoting a positive image of migrant workers and encouraging their pursuit of an improved quality of life can help alleviate social exclusion, strengthen their sense of identity, and foster deeper roots in cities. Furthermore, providing supportive platforms for returning migrant workers can enhance their active willingness to return home.

Abstract The widely tunable and high resolution mid-infrared laser based on a BaGa4Se7 (BGSe) optical parametric oscillator (OPO) was demonstrated. A wavelength tuning range of 2.76–4.64 μm and a wavelength tuning resolution of about 0.3 nm were obtained by a BGSe (56.3°, 0°) OPO, which was pumped by a 1064 nm laser. It is the narrowest reported wavelength tuning resolution for BGSe OPO, and was obtained by simultaneously controlling the angle and temperature of BGSe. Graphical Abstract

With the widespread adoption of ultrasound guidance, Seldinger puncture techniques, and intracardiac electrical positioning technology for the placement of peripherally inserted central catheters in recent years, an increasing number of medical staff and patients now accept peripheral placement of totally implantable venous access devices (TIVADs) in the upper arm. This approach has the advantage of completely avoiding the risks of hemothorax, pneumothorax, and neck and chest scarring. Medical specialties presently engaged in this study in China include internal medicine, surgery, anesthesiology, and interventional departments. However, command over implantation techniques, treatment of complications, and proper use and maintenance of TIVAD remain uneven among different medical units. Moreover, currently, there are no established quality control standards for implantation techniques or specifications for handling complications. Thus, this expert consensus is proposed to improve the success rate of TIVAD implantation via the upper-arm approach, reduce complication rates, and ensure patient safety. This consensus elaborates on the technical indications and contraindications, procedures and technical points, treatment of complications, and the use and maintenance of upper-arm TIVAD, thus providing a practical reference for medical staff.

Abstract Barium titanate (BaTiO3, BT) was co‐doped by solid‐state sintering with niobium pentoxide (Nb2O5) and cobalt trioxide (Co3O4) as dopants. The modified barium titanate containing Nb and Co (BTNC) with larger particle size (0.5–1 μm) and silver powder (Ag) with smaller particle size (25 nm) were co‐filled with polyvinylidene fluoride (PVDF) to prepare (BTNC‐Ag)/PVDF three‐phase composites. The morphology and crystal structure of composites were characterised by scanning electron microscope (SEM) and X‐ray diffraction (XRD), respectively. SEM shows that when the volume ratio of BTNC and Ag in the composite is 4:1, the two fillers have good dispersion in polymer matrix and could intersperse with each other to reduce voids. XRD patterns display that the filling of BTNC and Ag powders was conducive to promoting the enhancement of the diffraction peaks of β phase and γ phase in PVDF. The dielectric properties of the composites are effectively enhanced through the synergistic effect of the micro‐nano bicomponent ceramic BTNC and conductive particles Ag co‐filled polymer PVDF. When the volume ratio of filler (BTNC:Ag = 4:1) to matrix PVDF is 2/1, the dielectric properties of the composite are the best, the dielectric constant reaches 134.1 at 102 Hz and the dielectric loss is 0.04.

Abstract Despite a recent line of research highlighting trust as an important determinant of residents’ happiness in Western countries, empirical evidence regarding the strength of these linkages in the developing world needs to be more comprehensive and conclusive. This paper contributes to this literature by performing a deeper examination into the trust-based explanation of happiness and, particularly, exploring the mediating role of social capital in rural China, where rapid economic growth coexists with gradual and fundamental social changes. Using data from the nationally representative cross-sectional Chinese General Social Survey (CGSS) in 2012, 2013, and 2015, we indicate that trust positively affects the happiness of rural residents and social capital. Our findings support the role of social capital in mediating happiness prediction. Furthermore, additional tests suggest heterogeneous social capital for different ages and degrees of regional market competition. Specifically, the mediating effect of social capital on rural residents over 30 years old (inclusive) and living in high-market competition areas is significant. These findings provide a valuable direction for the government that producing an environment that enhances social networks and facilitates the exchange of social support holds promise for improving the life satisfaction of the rural Chinese population. Trust can significantly improve rural residents’ happiness through increasing social capital. The government should effectively manage the rural social trust crisis and help rural residents build social capital in multiple ways, improving the life satisfaction of rural residents.

Ultralight power generation equipment has high requirements for the power density, continuous operation, and transient stability of the whole machine, and there is a direct conflict between high power density and substantial stability control in the power unit design. In order to meet the power density requirements of ultralight power stations, three main problems need to be solved, namely engine speed oscillation and flameout in the process of load power mutation, matching of the generator and engine torque, and stabilizing the voltage waveform of the AC output end. In this paper, we proposed an exquisite compound control strategy for ultralight power generation systems in engines, generators, and inverters. The effectiveness and practicability of the proposed strategy are verified by both simulation and experiment. The results show that the proposed control strategy can effectively solve the instability problem of the ultralight generator set and improve the stability of the system, where response recovery can be achieved within 0.9 s under the conditions of a total load increase or decrease, and the mismatching degree of the generator following the engine is reduced by 90%. The strategy could also guarantee long-term stable operation with high-quality electrical energy output.

A wide-speed, ultra-light power generation system is a critical power generation unit structure, often because of its high efficiency and power density. Lightness and reliability are two key design indicators within the system, albeit they could lead to contradictory problems, particularly in systems containing prime movers, batteries, generators, rectifiers, and inverters. Ultra-light generator sets are facing more severe problems and contradictions in designing in terms of matching, coordinating, and stabilizing the components in the systems. This paper describes the system design of a low-cost and high-reliability microgenerator set: a gasoline engine, three-phase permanent magnet synchronous generator, rectifier, and inverter. Moreover, the matching relationship between the four parts and the design effect of each part of the power generation system was analyzed, simulated, and tested to verify the effectiveness and feasibility of the so-designed system.

ABSTRACT: Genotype imputation is a powerful technique employed by next-generation sequencing (NGS) and genotyping arrays, which can significantly enhance the cost-effectiveness and efficiency of genomic selection. The accuracy of imputation is largely determined by the choice of reference panel, with previous studies generally demonstrating that a closely related population as a reference panel leads to greater accuracy than a more distantly related population. Various strategies have been proposed for selecting desirable individuals via targeted resequencing, but their efficiencies need further improvement. In this study, we present a practical broiler selection methodology for a local Chinese chicken line that integrates established methods based on pedigree, genomics, and random sampling, and leverages genotype and pedigree information from the yellow-plumage dwarf chicken line. The efficacy of these selection strategies was assessed by evaluating their ability to accurately impute masked genotypes from data obtained using a 600K chip. Our findings reveal that the pedigree-based method yields superior accuracy in genotype imputation, whereas the haplotype-based method exhibits greater stability. Nonetheless, the impact of these targeted methods for selecting key individuals is slightly different when initiating a new sequencing project in a production context. Overall, this study highlights the advantages of using the pedigree-based approach as the preferred method for optimizing genotype imputation in broiler chickens.

Objective: To explore the relationships among self-perceived burden, resourcefulness and depression, and to study further whether resourcefulness mediates the effects of self-perceived burden on depression in young and middle-aged stroke patients. Methods: A cross-sectional survey was conducted with 1050 young and middle-aged stroke patients. We used a general demographic questionnaire, Self-Perceived Burden Scale (SPBS), Resourcefulness Scale© (RS) and Hamilton Depression Scale (HAMD) to assess self-perceived burden, resourcefulness, and depression. Statistical methods included correlation analysis, multiple linear regression, and structural equation model. Results: 1018 valid questionnaires were collected with a response rate of 96.95%. Resourcefulness was inversely correlated with self-perceived burden (r = −0.367, p

Abstract Background Skeletal muscle is comprised of heterogeneous myofibers that differ in their physiological and metabolic parameters. Of these, slow-twitch (type I; oxidative) myofibers have more myoglobin, more mitochondria, and higher activity of oxidative metabolic enzymes compared to fast-twitch (type II; glycolytic) myofibers. Methods In our previous study, we found a novel LncRNA-TBP (for “LncRNA directly binds TBP transcription factor”) is specifically enriched in the soleus (which has a higher proportion of slow myofibers). The primary myoblast cells and animal model were used to assess the biological function of the LncRNA-TBP in vitro or in vivo. Meanwhile, we performed a RNA immunoprecipitation (RIP) and pull-down analysis to validate this interaction between LncRNA-TBP and TBP. Results Functional studies demonstrated that LncRNA-TBP inhibits myoblast proliferation but promotes myogenic differentiation in vitro. In vivo, LncRNA-TBP reduces fat deposition, activating slow-twitch muscle phenotype and inducing muscle hypertrophy. Mechanistically, LncRNA-TBP acts as a regulatory RNA that directly interacts with TBP protein to regulate the transcriptional activity of TBP-target genes (such as KLF4, GPI, TNNI2, and CDKN1A). Conclusion Our findings present a novel model about the regulation of LncRNA-TBP, which can regulate the transcriptional activity of TBP-target genes by recruiting TBP protein, thus modulating myogenesis progression and inducing slow-twitch fibers. Video Abstract

ABSTRACT: Genomic selection using single nucleotide polymorphism (SNP) markers is now intensively investigated in breeding and has been widely utilized for genetic improvement. Currently, several studies have used haplotype (consisting of multiallelic SNPs) for genomic prediction and revealed its performance advantage. In this study, we comprehensively evaluated the performance of haplotype models for genomic prediction in 15 traits, including 6 growth, 5 carcass, and 4 feeding traits in a Chinese yellow-feathered chicken population. We adopted 3 methods to define haplotypes from high-density SNP panels, and our strategy included combining Kyoto Encyclopedia of Genes and Genomes pathway information and considering linkage disequilibrium (LD) information. Our results showed an increase in prediction accuracy due to haplotypes ranging from -0.04∼27.16% in all traits, where the significant improvements were found in 12 traits. The estimates of haplotype epistasis heritability were strongly correlated with the accuracy increase by haplotype models. In addition, incorporating genomic annotation information could further increase the accuracy of the haplotype model, where the further increase in accuracy is significantly relative to the increase of relative haplotype epistasis heritability. The genomic prediction using LD information for constructing haplotypes has the best prediction performance among the 4 traits. These results uncovered that haplotype methods were beneficial for genomic prediction, and the accuracy could be further increased by incorporating genomic annotation information. Moreover, using LD information would potentially improve the performance of genomic prediction.

ABSTRACT: In a previous study, the laying pattern of Muscovy duck was explored by macro-fitting the laying curve of Muscovy duck, and transcriptome sequencing technique of the ovarian tissues was used to screen the egg-related gene “TAT.” Moreover, recent results have shown that TAT is expressed in organs such as oviduct, ovary, and testis. The objective of this study is to examine the effect of TAT gene on egg production traits of Muscovy ducks. First, the expression levels of TAT gene in highest producing (HP) and lowest producing (LP) in 3 tissues related to reproduction were examined, and the results indicated that the expression of TAT gene in hypothalamus was significantly different between HP and LP groups. Then, 6 SNP loci (g. 120G>T, g, 122G>A, g, 254G> A, g. 270C >T, g, 312G>A, and g. 341C>A) were detected in TAT gene. Further, association analysis between the six SNP loci of TAT gene and egg production traits of 652 individual Muscovy ducks was done. The results showed that g. 254G>A and g. 270C>T were significantly correlated (P < 0.05 or 0.001) with the egg production traits of Muscovy ducks. This study elucidated the molecular mechanism that TAT gene might be regulating the egg production traits of Muscovy ducks.

Neoadjuvant systemic therapy (NST) is widely applied in breast cancer treatment, but individuals respond differently to the same NST regimen. It is unclear which patients should adjust their NST regimen and what such an adjustment should be, especially for patients with radiologically partial response (PR). This study aimed to identify a quantitative efficacy evaluation index to evaluate the therapeutic effect of NST. 164 patients were enrolled in this study received four cycles of epirubicin and cyclophosphamide (EC), followed by four cycles of taxanes with trastuzumab [T(H)], if needed. Of patients with a volume change rate of EC treatment (δV1) below 0.80, more than half benefited from subsequent T(H) treatment compared with EC treatment. Importantly, for δV1 of 0.80 and higher, patients’ subsequent T(H) treatment was not as efficient as previous EC treatment and they have a lower pathological complete response (pCR) rate. Across all patients, nanoparticle albumin-bound paclitaxel had a numerically higher pCR rate over other taxanes in patients with triple-negative breast cancer. This study showed that the volume change rate is better than the diameter change rate in monitoring the therapeutic effect of NST. Furthermore, δV1 is a good quantitative efficacy evaluation index to distinguish patients resistant to EC treatment and predict the pCR rate and guide the adjustment of individualized NST regimens.

Abstract Skeletal muscle is the largest metabolic organ in the body, and its metabolic flexibility is essential for maintaining systemic energy homeostasis. Metabolic inflexibility in muscles is a dominant cause of various metabolic disorders, impeding muscle development. In our previous study, we found lncRNA ZFP36L2-AS (for “ZFP36L2-antisense transcript”) is specifically enriched in skeletal muscle. Here, we report that ZFP36L2-AS is upregulated during myogenic differentiation, and highly expressed in breast and leg muscle. In vitro, ZFP36L2-AS inhibits myoblast proliferation but promotes myoblast differentiation. In vivo, ZFP36L2-AS facilitates intramuscular fat deposition, as well as activates fast-twitch muscle phenotype and induces muscle atrophy. Mechanistically, ZFP36L2-AS interacts with acetyl-CoA carboxylase alpha (ACACA) and pyruvate carboxylase (PC) to induce ACACA dephosphorylation and damaged PC protein stability, thus modulating muscle metabolism. Meanwhile, ZFP36L2-AS can activate ACACA to reduce acetyl-CoA content, which enhances the inhibition of PC activity. Our findings present a novel model about the regulation of lncRNA on muscle metabolism.

A widely tunable 4.16–7.31 μm high-resolution mid-infrared optical parametric oscillator with a type-I BaGa4Se7 (BGSe) crystal pumped by a 1064 nm laser was demonstrated for the first time. We have achieved a wavelength tuning accuracy of less than 5 nm through simultaneously performing temperature tuning and angular tuning on the BGSe (46°, 0°) Optical Parametric Oscillation (OPO). The wavelength resolution will be less than 1 nm when using commercial temperature control equipment.

ABSTRACT: Prior studies on transcriptomes of hypothalamus and ovary revealed that AKT3 is one of the candidate genes that might affect egg production in White Muscovy ducks. The role of AKT3 in the uterus during reproductive processes cannot be overemphasized. However, functional role of this gene in the tissues and on egg production traits of Muscovy ducks remains unknown. To identify the relationship between AKT3 and egg production traits in ducks, relative expression profile was first examined prior to identifying the variants within AKT3 that may underscore egg production traits [age at first egg (AFE), number of eggs at 300 d (N300D), and number of eggs at 59 wk (N59W)] in 549 ducks. The mRNA expression of AKT3 gene in high producing (HP) ducks was significantly higher than low producing (LP) ducks in the ovary, oviduct, and hypothalamus (P < 0.05 or 0.001). Three variants in AKT3 (C-3631A, C-3766T, and C-3953T) and high linkage block between C-3766T and C-3953T which are significantly (P < 0.05) associated with N300D and N59W were discovered. This study elucidates novel knowledge on the molecular mechanism of AKT3 that might be regulating egg production traits in Muscovy ducks.

ABSTRACT: Background: Livestock and poultry skeletal muscle development was regulated by a variety of genetic factors playing an essential role in this process. In our de novo sequencing, a missense mutation in ISPD exon 2 showed a selected signature for well-developed muscles. However, the relationship between this mutation and muscle phenotypes remains unclear. Results: Based on the genotype bias of this missense mutation in gamecock chickens, we compared the cross-section of muscle fibers among the individuals with different genotypes with the results showing a genotype preference of this missense mutation in the chickens with hypertrophic muscle fibers. Comparing TT- and CC-genotype ISPD showed that TT- genotype was associated with heavier skeletal muscle and a better effect on myogenesis and myofiber stability. At cellular level, ISPD was identified to inhibit myoblast proliferation, differentiation and antagonize of muscular atrophy. Conclusion: It can be concluded that the missense mutation of Arg84Lys in ISPD was associated with well-developed muscles and demonstrated to improve chicken muscles’ development.

Yellow-feathered chickens have great nutritional value and are widely and traditionally used in China, on an industrial scale as broilers. The presence of intestinal microbes has been shown to correlate with poultry performance and serves as an essential reservoir of antibiotic resistance genes (ARGs). Antibiotic resistance is a major public health concern. Here, we investigated functional characteristics of the gut microbiome of indigenous Chinese yellow-feathered chickens (the Huiyang Bearded, Xinghua, Huaixiang, Zhongshan Shanlan, Qingyuan Partridge, and Yangshan chickens) through metagenomic sequencing and reconstructed 409 draft genomes, including 60 novel species and 6 novel genera. Furthermore, we assessed the functions of the intestinal microbial communities and examined the ARGs within them. The results showed that the microbial populations of yellow-feathered broilers were primarily dominated by Bacteroidetes and Firmicutes at the phylum level and Bacteroides at the genus level. Furthermore, the Qingyuan Partridge chicken showed a significantly higher abundance of Prevotella than the other five breeds of chicken. Principal coordinates analysis indicated significant differences in the structures of microbial communities and ARGs, based on the binary Jaccard distance, among the six chicken breeds. Moreover, 989 ARGs conferring tetracycline, multidrug, and aminoglycoside resistance were identified, which represented more than 80% of the faecal resistomes; the most abundant gene in the yellow-feathered chickens was tet(Q). In addition, we found the greatest abundance of resistance genes in Xinghua chickens, indicating that Xinghua chickens are highly resistant to antibiotics. Overall, our findings revealed differences in the gut microbial community structure of indigenous Chinese yellow-feathered broiler breeds and the composition and characteristics of ARGs and antibiotic resistance that enabled us to reconstruct the yellow-feathered chicken gut microbial community genomes. The current data significantly improves our knowledge of the gut microbiome and antibiotic resistance of popular broiler breeds in China.

We report on a high-power acoustic-optical (AO) Q-switched Tm:YLF laser operating at ~1.83 μm by controlling the transmittance of the output coupler. Under the continuous-wave (CW) operation, the maximum output power of 13 W is achieved, and the slope efficiency is up to 32.7%. With a YAG etalon inserted into the cavity, the linewidth is compressed below 0.5 nm with a maximum output power of 12.2 W. In the Q-switched state, the maximum pulsed output power of 10.32 W is achieved with a pulse duration of 150 ns when the repetition rate is 15 kHz. And the maximum pulsed energy of 1.13 mJ is generated with a duration of 131 ns at 5 kHz. As far as we know, this is the highest output power reported for the CW and pulsed 1.83 μm laser. In addition, the relationship between the output wavelength and crystal length is theoretically analyzed, which shows that increasing the loss of 1880 nm is the key to high-power 1.83 μm laser output.

BackgroundImproving the health self-management level of patients with tuberculosis (TB) is significant for reducing drug resistance, improving the cure rate, and controlling the prevalence of TB. Mobile health (mHealth) interventions based on behavioral science theories may be promising to achieve this goal. ObjectiveThis study aims to explore and conduct an mHealth intervention based on the Integrated Theory of Health Behavior Change (ITHBC) in patients with pulmonary TB to increase their ability of self-care management. MethodsA prospective randomized controlled study was conducted from May to November 2020. A total of 114 patients who were admitted consecutively to the TB clinic of Harbin Chest Hospital, China from May 2020 to August 2020 were recruited by convenience sampling. Patients were divided into the control group and intervention group, and all received a 3-month intervention. Patients in the intervention group and the control group received routine medical and nursing care in the TB clinic, including the supervision of their medications. In addition, pharmacist-assisted mHealth (WeChat) intervention based on the ITHBC theory about TB management was provided to the intervention group. The primary outcome was self-management behavior, while the secondary outcomes were TB awareness, self-efficacy, social support, and degree of satisfaction with health education. The outcomes were measured using web-based self-designed and standard questionnaires administered at baseline and at the end point of the study. Intergroup data were assessed using the Mann-Whitney U test, whereas intragroup data were assessed with the Wilcoxon test (for paired samples). ResultsA total of 112 patients (59 in intervention group and 53 in control group) completed the study. After the intervention, a statistically significant increase was noted in the scores of each item of self-care management behaviors compared with the scores at the baseline (P

PurposePrescription refills are long-term prescriptions for chronic patients in stable status, which varies from country to country. A well-established prescription refill system is beneficial for chronic patients’ medication management and facilitates the efficacy of clinical care. Therefore, we carried out a bibliometric analysis to examine the development of this field.SummaryPublications on prescription refills from 1970 to 2021 were collected in the Web of Science Core Collection (WoSCC). Search strategy TS = “prescri* refill*” OR “medi* refill*” OR “repeat prescri*” OR “repeat dispens*” OR TI = refill* was used for search. VOSviewer was applied to visualize the bibliometric analysis. A total of 319 publications were found in WoSCC. Study attention on prescription refills has shown a steady rise but is still low in recent years. The United States was the most productive country, which had the highest total citations, average citations per publication, and the highest H-index, and participated in international collaboration most frequently. The University of California system was the most productive institution. The U.S. Department of Veterans Affairs was the institution with the most citations, most average citation, and highest H-index. Sundell was the most productive author, and Steiner J. F. was the most influential author. “Adherence,” “medication,” and “therapy” were the most prominent keywords.ConclusionPublications on prescription refills have increased rapidly and continue to grow. The United States had the leading position in the area. It is recommended to pay closer attention to the latest hotspots, such as “Opioids,” “Surgery,” “Differentiated care,” and “HIV.”

As the world population grows, muscle atrophy leading to muscle wasting could become a bigger risk. Long noncoding RNAs (lncRNAs) are known to play important roles in muscle growth and muscle atrophy. Meanwhile, it has recently come to light that many putative small open reading frames (sORFs) are hidden in lncRNAs; however, their translational capabilities and functions remain unclear. In this study, we uncovered 104 myogenic-associated lncRNAs translated, in at least a small peptide, by integrated transcriptome and proteomic analyses. Furthermore, an upstream ORF (uORF) regulatory network was constructed, and a novel muscle atrophy-associated lncRNA named SMUL (Smad ubiquitin regulatory factor 2 [SMURF2] upstream lncRNA) was identified. SMUL was highly expressed in skeletal muscle, and its expression level was downregulated during myoblast differentiation. SMUL promoted myoblast proliferation and suppressed differentiation in vitro. In vivo, SMUL induced skeletal muscle atrophy and promoted a switch from slow-twitch to fast-twitch fibers. In the meantime, translation of the SMUL sORF disrupted the stability of SMURF2 mRNA. Mechanistically, SMUL restrained SMURF2 production via nonsense-mediated mRNA decay (NMD), participating in the regulation of the transforming growth factor β (TGF-β)/SMAD pathway and further regulating myogenesis and muscle atrophy. Taken together, these results suggest that SMUL could be a novel therapeutic target for muscle atrophy.

Amid the epoch of global overpopulation, the agroforestry system can intervene as a novel practice that can safeguard agricultural sustainability, provide a means of livelihood, yield ecological benefits, and contribute to household food security. However, the adoption of sustained agroforestry practices requires an understanding of both farmers’ personal characteristics and perceived statuses, constituting a difficult task to anticipate, analyze, and visualize. To this end, it is crucial to understand and identify the most significant factors driving the adoption of agroforestry. This research attempts to examine the determinants and psychological drivers of smallholder farmers’ intention to plant trees on farmland and the factors that may discourage them from doing so. The conceptual framework of the study was developed based on the theory of planned behavior. We draw on survey data from 400 smallholder farmers in a northern irrigated plain of Pakistan. A binary logistic regression model was employed to explore how socio-psychological drivers influence farmers’ decision to adopt agroforestry practices. The study results reveal that 60.5% of the respondents prefer to adopt agroforestry practices. The results from a logit estimation showed that socio-economic variables such as family size, land ownership, subsidies, livestock rearing, sources of energy, and total income had a significant positive influence on the planting of trees on farmland, while age had a negative influence. Moreover, runoff control and the control of heat significantly affect the perceptions of farmers regarding the adoption of agroforestry. The findings suggest that implementing policies that enhance the delivery of robust agricultural extension services and training programs for farmers could disseminate the agroforestry system countrywide, which might offer substantial benefits for farmers in the long term while maintaining environmental integrity.

Background: The Duke Anticoagulation Satisfaction Scale (DASS) offers a method to measure the quality of life and satisfaction of patients taking oral anticoagulants. The study aimed to validate the Chinese version of the DASS in Chinese patients on anticoagulation therapy.Methods: The DASS was translated, back-translated, and transculturally adapted into the Chinese version and then administered to participants taking oral anticoagulants in a physician–pharmacist collaborative anticoagulation clinic at a tertiary teaching hospital from October 2019 to December 2020. Reliability was analyzed through Cronbach’s alpha (α) and split-half reliability. Confirmatory factor analysis was performed to test the structural validity of the scale. Exploratory factor analysis was performed for items in the scales using the varimax rotation method.Results: A total of 189 patients completed the Chinese version of the DASS. Four dimensions and 23 items were included, with Cronbach’s α values of 0.89, 0.81, 0.89, and 0.74 for limitations on physical activities, diet restrictions, hassles and burdens, and positive psychological effect, respectively. Cronbach’s α coefficient of whole scale was 0.91. The split-half reliability of this scale is 0.747 (>0.7).Conclusion: The Chinese version of the DASS indicated excellent reliability and validity, compared to the original version. It could provide a practical instrument for healthcare practitioners to evaluate satisfaction and quality of life for anticoagulated patients in China. The difference in quality of life between patients taking warfarin and novel oral anticoagulants (NOACs) needs to be further explored in future studies.

A high-power narrow-linewidth 1.55 μm pulsed laser, based on MgO:PPLN OPO, has been achieved using a F–P etalon. The pump source is a 1064 nm acousto-optical (AO) Q-switched Nd:YAG laser with a repetition rate of 10 kHz. Under the maximum pump power of 18 W, the signal output power of 2.57 W is demonstrated at 1551.1 nm with a linewidth of 0.07 nm, corresponding to a slope efficiency of 16.1%. Different from traditional inversion lasers, the narrow-linewidth wavelength tunability of approximately 1.55 μm can be realized by changing the temperature.

ABSTRACT: In China, the low egg production rate is a major challenge to Muscovy duck farmers. Hypothalamus and ovary play essential role in egg production of birds. However, there are little or no reports from these tissues to identify potential candidate genes responsible for egg production in White Muscovy ducks. A total of 1,537 laying ducks were raised; the egg production traits which include age at first egg (days), number of eggs at 300 d, and number of eggs at 59 wk were recorded. Moreover, 4 lowest (LP) and 4 highest producing (HP) were selected at 59 wk of age, respectively. To understand the mechanism of egg laying regulation, we sequenced the hypothalamus and ovary transcriptome profiles in LP and HP using RNA-Seq. The results showed that the number of eggs at 300 d and number of eggs at 59 wk in the HP were significantly more (P < 0.001) than the LP ducks. In total, 106.98G clean bases were generated from 16 libraries with an average of 6.68G clean bases for each library. Further analysis showed 569 and 2,259 differentially expressed genes (DEGs) were identified in the hypothalamus and ovary between LP and HP, respectively. The KEGG pathway enrichment analysis revealed 114 and 139 pathways in the hypothalamus and ovary, respectively which includes Calcium signaling pathway, ECM-receptor interaction, Focal adhesion, MAPK signaling pathway, Apoptosis and Apelin signaling pathways that are involved in egg production. Based on the GO terms and KEGG pathways results, 10 potential candidate genes (P2RX1, LPAR2, ADORA1, FN1, AKT3, ADCY5, ADCY8, MAP3K8, PXN, and PTTG1) were identified to be responsible for egg production. Further, protein-protein interaction was analyzed to show the relationship between these candidate genes. Therefore, this study provides useful information on transcriptome of hypothalamus and ovary of LP and HP Muscovy ducks.

(1) Background: Excessive abdominal fat deposition in broilers not only causes feed waste but also leads to a series of metabolic diseases. It has gradually become a new breeding goal of the broiler industry to improve growth rates and to reduce abdominal fat rates. In a previous study, PROS1 was highly expressed in low-abdominal fat broilers, suggesting a potential role in broilers adipogenesis. However, the function of PROS1 in preadipocytes and its association with abdominal fat traits need to be characterized. (2) Methods: qRT-PCR and Western Blot were used to quantify gene expression at the RNA and protein levels; flow cytometry and EdU were carried out to detect cell proliferation; and a GLM analysis was used to determine the association between PROS1 SNPs and carcass traits. (3) Results: PROS1 was downregulated in high-abdominal fat chicken; PROS1 contributed preadipocyte proliferation but suppressed preadipocyte differentiation; and the SNPs in the PROS1 5′ flank were significantly associated with the abdominal fat weight rate. (4) Conclusions: Chicken PROS1 is able to suppress adipogenesis, and its polymorphisms are associated with the abdominal fat weight rate, which can be considered the molecular markers for chicken breeding, indicating that PROS1 is an effective potential gene in regulating abdominal fat deposition.

It is widely acknowledged that object segmentation is a significant research field for computer vision and a key process for many other visual tasks. In the past unsupervised single-image segmentation, there are often cases where the segmentation result is not good. In the current supervised single-image segmentation, it is necessary to rely on a large number of data annotations and long-term training of the model. Then, people attempted to segment simultaneously the common regions from multiple images. On the one hand, it does not need to use a large amount of labeled data to train in advance. On the other hand, it utilizes the consistency constraint between images to better obtain the object information. This idea can generate better performance than the traditional one did, resulting in many methods related to object co-segmentation. This paper reviews some classic and effective object co-segmentation methods, including saliency-based approaches, joint-processing-based approaches, graph-based approaches, and others. For different methods, we select two or three related models to elaborate, such as a model based on random walks. Moreover, in order to exhibit and evaluate these methods objectively and comprehensively, we not only summarize them in the form of flowcharts and algorithm summaries, but also compare their performance with visualization methods and evaluation metrics, such as intersection-over-union, consistency error, and precision-recall rate. From the experiment, we also attempt to clarify and analyze the existing problems. Finally, we point out the challenges and directions and open new venues for future researchers in the field.

Recent advances in the corporate processing of images have exhibited its advantages over the individual processing. Image co-segmentation aims to segment shared objects from two or more relevant images, and is becoming more and more interesting for computer vision researchers. Many applications need accurate and efficient segmentation techniques: indoor navigation, autonomous driving, and virtual reality systems to name a few. Although numerous techniques have been proposed, it is still lacking a deep review of image co-segmentation techniques. In this paper, we provide a review on the fundamentals and challenges of image co-segmentation techniques. We organize the recent advances of image co-segmentation into seven major frameworks: graph based framework, clustering based framework, partial differential equation based framework, quadratic programming based framework, low rank matrix recovery based framework, joint optimization based framework, and machine learning based framework. We expect this review to be beneficial to both fresh and senior researchers in this field.

The immune cell inflammation response is closely related to the occurrence of disease, and much evidence has shown that circular RNAs (circRNAs) play vital roles in the occurrence of disease. However, the biological function and regulatory mechanisms of circRNAs in the immune cell inflammation response remain poorly understood. In this study, we constructed an inflammatory model using lipopolysaccharide (LPS)-stimulated chicken macrophage lines (also known as HD11) to verify the function and mechanism of the novel circDCLRE1C (ID: gga_circ_0001674), which was significantly upregulated in spleen tissues infected by coccidia and the macrophage cells exposed to LPS. The results showed that circDCLRE1C aggravated LPS-induced inflammation and apoptosis in HD11 cells. Systemically, circDCLRE1C acted as a sponge for miR-214b-3p binding sites thereby regulating the expression of STAT3. The overexpression of miR-214b-3p rescued the pro-inflammatory effect of circDCLRE1C in HD11 cells stimulated with LPS, and rescued the high expression of STAT3. In conclusion, our study showed that circDCLRE1C could aggravate LPS-induced inflammation and apoptosis through competitive adsorption of miR-214b-3p, thereby increasing the expression of STAT3.

Coccidiosis is a major hazard to the chicken industry, but the host’s immune response to coccidiosis remains unclear. Here, we performed Eimeria coccidia challenge in 28-day-old ROSS 308 broilers and selected the bursa from the three most severely affected individuals and three healthy individuals for RNA sequencing. We obtained 347 DEGs from RNA-seq and found that 7 upregulated DEGs were enriched in Cytokine-cytokine receptor interaction pathway. As the DEGs with the highest expression abundance in these 7 genes, TNFRSF6B was speculated to participate in the process of host’s immune response to coccidiosis. It is showed that TNFRSF6B can polarize macrophages to M1 subtype and promote inflammatory cytokines expression. In addition, the expression of TNFRSF6B suppressed HD11 cells apoptosis by downregulating Fas signal pathway. Besides, TNFRSF6B-mediated macrophages immunity activation can be reversed by apoptosis. Overall, our study indicates that TNFRSF6B upregulated in BAE, is capable of aggravating the inflammatory response by inhibiting macrophages apoptosis via downregulating Fas signal pathway, which may participate in host’s immune response to coccidiosis.

Keratinocytes are organised into a periodic pattern in feather branching, but how this is regulated is unclear. Here, the authors show that there is a coordinated change in cell shape and adherence, mediated by Notch, FGF signalling and Rho GTPases, which in turn regulates feather branching.

Background/Aims: Earlobe color is a typical external trait in chicken. There are some previous studies showing that the chicken white/red earlobe color is a polygenic and sex-linked trait in some breeds, but its molecular genetic and histological mechanisms still remain unclear. Methods: We herein utilized histological section, genome-wide association study (GWAS) and RNA-seq, further to investigate the potential histological and molecular genetic mechanisms of white/red earlobe formation in Qiangyuan Partridge chicken (QYP). Results: through histological section analysis, we found the dermal papillary layer of red earlobes had many more blood vessels than that of white earlobes. And we identified a total of 44 SNPs from Chromosome 1, 2, 3, 4, 9, 10, 11, 13, 19, 20, 23 and Z, that was significantly associated with the chicken white/red earlobe color from GWAS, along with 73 significantly associated genes obtained (e.g., PIK3CB, B4GALT1 and TP63), supporting the fact that the white/red earlobe color was also polygenic and sex-linked in QYP. Importantly, PIK3CB and B4GALT1 are both involved in the biological process of angiogenesis, which may directly give rise to the chicken white earlobe formation through regulating blood vessel density in chicken earlobe. Additionally, through contrast of RNA-seq profiles between white earlobe skins and red earlobe skins, we further identified TP63 and CDH1 differentially expressed. Combined with the existing knowledge of TP63 in epithelial development and tumor angiogenesis, we propose that down-regulated TP63 in white earlobes may play roles in thickening the skin and decreasing the vessel numbers in dermal papillary layer, thereby contributing to the white earlobe formation via paling the redness of the skin in QYP, but the specific mechanism remains to be further clarified. Conclusion: our findings advance the existing understanding of the white earlobe formation, as well as provide new clues to understand the molecular mechanism of chicken white/red earlobe color formation.

As the miniaturization of electronic appliances and microprocessors progresses, low-permittivity interlayer materials are becoming increasingly important for their suppression of electronic crosstalk, signal propagation delay and loss, and so forth. Herein, a kind of copolyimide (CPI) film with a “fluorene” rigid conjugated structure was prepared successfully. By introducing 9,9-Bis(3-fluoro-4-aminophenyl) fluorene as the rigid conjugated structure monomer, a series of CPI films with different molecular weights were fabricated by in situ polymerization, which not only achieved the reduction of permittivity but also maintained excellent thermodynamic stability. Moreover, the hydrophobicity of the CPI film was also improved with the increasing conjugated structure fraction. The lowest permittivity reached 2.53 at 106 Hz, while the thermal decomposition temperature (Td5%) was up to 530 °C, and the tensile strength was ≥ 96 MPa. Thus, the CPI films are potential dielectric materials for microelectronic and insulation applications.

Growth performance is an important economic trait in chicken. MicroRNAs (miRNAs) have been shown to play important roles in various biological processes, but their functions in chicken growth are not yet clear. To investigate the function of miRNAs in chicken growth, breast muscle tissues of the two-tail samples (highest and lowest body weight) from Recessive White Rock (WRR) and Xinghua Chickens (XH) were performed on high throughput small RNA deep sequencing. In this study, a total of 921 miRNAs were identified, including 733 known mature miRNAs and 188 novel miRNAs. There were 200, 279, 257 and 297 differentially expressed miRNAs in the comparisons of WRRh vs. WRRl, WRRh vs. XHh, WRRl vs. XHl, and XHh vs. XHl group, respectively. A total of 22 highly differentially expressed miRNAs (fold change > 2 or < 0.5; p-value < 0.05; q-value < 0.01), which also have abundant expression (read counts > 1000) were found in our comparisons. As far as two analyses (WRRh vs. WRRl, and XHh vs. XHl) are concerned, we found 80 common differentially expressed miRNAs, while 110 miRNAs were found in WRRh vs. XHh and WRRl vs. XHl. Furthermore, 26 common miRNAs were identified among all four comparisons. Four differentially expressed miRNAs (miR-223, miR-16, miR-205a and miR-222b-5p) were validated by quantitative real-time RT-PCR (qRT-PCR). Regulatory networks of interactions among miRNAs and their targets were constructed using integrative miRNA target-prediction and network-analysis. Growth hormone receptor (GHR) was confirmed as a target of miR-146b-3p by dual-luciferase assay and qPCR, indicating that miR-34c, miR-223, miR-146b-3p, miR-21 and miR-205a are key growth-related target genes in the network. These miRNAs are proposed as candidate miRNAs for future studies concerning miRNA-target function on regulation of chicken growth.

Accumulating studies report that microRNAs (miRNAs) are actively involved in skeletal myogenesis. Previously, our study revealed that miR-146b-3p was related to the growth of skeletal muscle. Here, we further report that miR-146b-3p is essential for the proliferation, differentiation, and apoptosis of chicken myoblast. Elevated expression of miR-146b-3p can dramatically suppress proliferation and differentiation, and facilitate apoptosis of chicken myoblast. Besides, we identified two target genes of miR-146b-3p, AKT1 and MDFIC, and found that miR-146b-3p can inhibit the PI3K/AKT pathway. Our study also showed that both AKT1 and MDFIC can promote the proliferation and differentiation while inhibit the apoptosis of myoblast in chicken. Overall, our results demonstrate that miR-146b-3p, directly suppressing PI3K/AKT pathway and MDFIC, acts in the proliferation, differentiation, and apoptosis of myoblast in chicken.

Circular RNAs and microRNAs widely exist in various species and play crucial roles in multiple biological processes. It is essential to study their roles in myogenesis. In our previous sequencing data, both miR-30a-3p and circular HIPK3 (circHIPK3) RNA, which are produced by the third exon of the HIPK3 gene, were differentially expressed among chicken skeletal muscles at 11 embryo age (E11), 16 embryo age (E16), and 1-day post-hatch (P1). Here, we investigated their potential roles in myogenesis. Proliferation experiment showed that miR-30a-3p could inhibit the proliferation of myoblast. Through dual-luciferase assay and Myosin heavy chain (MYHC) immunofluorescence, we found that miR-30a-3p could inhibit the differentiation of myoblast by binding to Myocyte Enhancer Factor 2 C (MEF2C), which could promote the differentiation of myoblast. Then, we found that circHIPK3 could act as a sponge of miR-30a-3p and exerted a counteractive effect of miR-30a-3p by promoting the proliferation and differentiation of myoblasts. Taking together, our data suggested that circHIPK3 could promote the chicken embryonic skeletal muscle development by sponging miR-30a-3p.

MicoRNAs (miRNAs), usually as gene regulators, participate in various biological processes, including stress responses. The hypothalamus–pituitary–adrenal axis (HPA axis) is an important pathway in regulating stress response. Although the mechanism that HPA axis regulates stress response has been basically revealed, the knowledge that miRNAs regulate stress response within HPA axis, still remains poor. The object of this study was to investigate the miRNAs in the pituitary and adrenal cortex that regulate chronic stress response with high-throughput sequencing. The pituitary and adrenal cortex of beagles and Chinese Field dogs (CFD) from a stress exposure group (including beagle pituitary 1 (BP1), CFD pituitary 1 (CFDP1), beagle adrenal cortex 1 (BAC1), CFD adrenal cortex 1 (CFDAC1)) and a control group (including beagle pituitary 2 (BP2), CFD pituitary 2 (CFDP2), beagle adrenal cortex 2 (BAC2), CFD adrenal cortex 2 (CFDAC2)), were selected for miRNA-seq comparisons. Comparisons, that were made in pituitary (including BP1 vs. BP2, CFDP1 vs. CFDP2, BP1 vs. CFDP1 and BP2 vs. CFDP2) and adrenal cortex (including BAC1 vs. BAC2, CFDAC1 vs. CFDAC2, BAC1 vs. CFDAC1 and BAC2 vs. CFDAC2), showed that a total of 39 and 18 common differentially expressed miRNAs (DE-miRNAs) (Total read counts > 1,000, Fold change > 2 & p-value < 0.001), that shared in at least two pituitary comparisons and at least two adrenal cortex comparisons, were detected separately. These identified DE-miRNAs were predicted for target genes, thus resulting in 3,959 and 4,010 target genes in pituitary and adrenal cortex, respectively. Further, 105 and 10 differentially expressed genes (DEGs) (Fold change > 2 & p-value < 0.05) from those target genes in pituitary and adrenal cortex were obtained separately, in combination with our previous corresponding transcriptome study. Meanwhile, in line with that miRNAs usually negatively regulated their target genes and the dual luciferase reporter assay, we finally identified cfa-miR-205 might play an important role by upregulating MMD in pituitary and hippocampus, thus enhancing the immune response, under chronic stress exposure. Our results shed light on the miRNA expression profiles in the pituitary and adrenal cortex with and without chronic stress exposure, and provide a new insight into miR-205 with its feasible role in regulating chronic stress in the pituitary and hippocampus through targeting MMD.

The geodesic active contour (GAC) model has been widely used due to the high precision of edge detection and the continuity of boundaries. However, compared to the gradient vector flow (GVF) field, it usually has poor performance in the object boundaries with concave shape. In this paper, the improvements that integrate the good performance of the GAC model and the GVF field are described in detail and the GAC_GVF&B model combining the GAC model with the GVF field and a balloon force is proposed. The experimental results demonstrate that the proposed method is insensitive to the positions of initial curves and can segment complex-shape or multi-objects images correctly with higher convergence.

PDE-based image segmentation based on the active contour model attracts many researchers due to its high precision of edge detection and the continuity of boundaries. Its basic idea is to define an energy functional on a dynamic curve which achieves its minimum when the curve conforms to the boundary of the objects. Thus, the image segmentation problem is in essence an optimization problem. The most widely used optimization method is the gradient descent method in PDE-based image segmentation. However, the convergence of the gradient descent method is very poor. In this paper, a quasi-Newton method is extended to the generalized quasi-Newton method, and then the generalized Newton method and the generalized quasi-Newton method are used to solve a simple region-based model and compared with the gradient decent method. Experimental results show that the generalized quasi-Newton method has accurate segmentation results in the least possible number of iteration. Moreover it is able to segment noisy images correctly.

Chicken growth performance provides direct economic benefits to the poultry industry. However, the underlying genetic mechanisms are unclear. The objective of this study was to identify candidate genes associated with chicken growth and investigate their potential mechanisms. We used RNA-Seq to study the breast muscle transcriptome in high and low tails of Recessive White Rock (WRRh, WRRl) and Xinghua chickens (XHh, XHl). A total of 60, 23, 153 and 359 differentially expressed genes were detected in WRRh vs. WRRl, XHh vs. XHl, WRRh vs. XHh and WRRl vs. XHl, respectively. GO, KEGG pathway and gene network analyses showed that CEBPB, FBXO32, FOXO3 and MYOD1 played key roles in growth. The functions of FBXO32 and FOXO3 were validated. FBXO32 was predominantly expressed in leg muscle, heart and breast muscle. After decreased FBXO32 expression, growth-related genes such as PDK4, IGF2R and IGF2BP3 were significantly down-regulated (P < 0.05). FBXO32 was significantly (P < 0.05) associated with carcass and meat quality traits, but not growth traits. FOXO3 was predominantly expressed in breast and leg muscle. In both of these tissues, the FOXO3 mRNA level in XH was significantly higher than that in WRR chickens with normal body weight (P < 0.05). In DF-1 cells, siRNA knockdown of FOXO3 significantly (P < 0.01) inhibited the MYOD expression and significantly up-regulated (P < 0.01 or P < 0.05) the expression of growth-related genes including CEBPB, FBXO32, GH, GHR, IGF1R, IGF2R, IGF2BP1, IGF2BP3, INSR, PDK1 and PDK4. Moreover, 18 SNPs were identified in FOXO3. G66716193A was significantly (P < 0.05) associated with growth traits. The sites C66716002T, C66716195T and A66716179G were significantly (P < 0.05) associated with growth or carcass traits. These results demonstrated that FOXO3 is a candidate gene influencing chicken growth. Our observations provide new clues to understand the molecular basis of chicken growth.

Runting and stunting syndrome (RSS), which is characterized by lower body weight, widely occurs in broilers. Some RSS chickens simply exhibit slow growth without pathological changes. An increasing number of studies indicate that broiler strains differ in susceptibility to infectious diseases, most likely due to their genetic differences. The objective of this study was to detect the differentially expressed miRNAs and mRNAs in RSS and normal chickens. By integrating miRNA with mRNA expression profiling, potential molecular mechanisms involved in RSS could be further explored. Twenty-two known miRNAs and 1,159 genes were differentially expressed in RSS chickens compared with normal chickens (P < 0.05). qPCR validation results displayed similar patterns. The differentially expressed genes were primarily involved in energy metabolism pathways. The antisense transcripts were extensively expressed in chicken liver albeit with reduced abundance. Dual-luciferase reporter assay indicated that gga-miR-30b/c directly target CARS through binding to its 3'UTR. The miR-30b/c: CARS regulation mainly occurred in liver. In thigh muscle and the hypothalamus, miR-30b/c are expressed at higher levels in RSS chickens compared with normal chickens from 2 to 6 w of age, and notably significant differences are observed at 4 w of age.

INTRODUCTION: Growth traits are important in poultry production, however, little is known for its regulatory mechanism at epigenetic level. Therefore, in this study, we aim to compare DNA methylation profiles between fast- and slow-growing broilers in order to identify candidate genes for chicken growth. Methylated DNA immunoprecipitation-sequencing (MeDIP-seq) was used to investigate the genome-wide DNA methylation pattern in high and low tails of Recessive White Rock (WRR(h); WRR(l)) and that of Xinhua Chickens (XH(h); XH(l)) at 7 weeks of age. The results showed that the average methylation density was the lowest in CGIs followed by promoters. Within the gene body, the methylation density of introns was higher than that of UTRs and exons. Moreover, different methylation levels were observed in different repeat types with the highest in LINE/CR1. Methylated CGIs were prominently distributed in the intergenic regions and were enriched in the size ranging 200-300 bp. In total 13,294 methylated genes were found in four samples, including 4,085 differentially methylated genes of WRR(h) Vs. WRR(l), 5,599 of XH(h) Vs. XH(l), 4,204 of WRR(h) Vs. XH(h), as well as 7,301 of WRR(l) Vs. XH(l). Moreover, 132 differentially methylated genes related to growth and metabolism were observed in both inner contrasts (WRR(h) Vs. WRR(l) and XH(h) Vs. XH(l)), whereas 129 differentially methylated genes related to growth and metabolism were found in both across-breed contrasts (WRR(h) Vs. XH(h) and WRR(l) Vs. XH(l)). Further analysis showed that overall 75 genes exhibited altered DNA methylation in all four contrasts, which included some well-known growth factors of IGF1R, FGF12, FGF14, FGF18, FGFR2, and FGFR3. In addition, we validate the MeDIP-seq results by bisulfite sequencing in some regions. CONCLUSIONS: This study revealed the global DNA methylation pattern of chicken muscle, and identified candidate genes that potentially regulate muscle development at 7 weeks of age at methylation level.

BACKGROUND: Abundant evidence indicates that chicken reproduction is strictly regulated by the hypothalamic-pituitary-gonad (HPG) axis, and the genes included in the HPG axis have been studied extensively. However, the question remains as to whether any other genes outside of the HPG system are involved in regulating chicken reproduction. The present study was aimed to identify, on a genome-wide level, novel genes associated with chicken reproductive traits. METHODOLOGY/PRINCIPAL FINDING: Suppressive subtractive hybridization (SSH), genome-wide association study (GWAS), and gene-centric GWAS were used to identify novel genes underlying chicken reproduction. Single marker-trait association analysis with a large population and allelic frequency spectrum analysis were used to confirm the effects of candidate genes. Using two full-sib Ningdu Sanhuang (NDH) chickens, GARNL1 was identified as a candidate gene involved in chicken broodiness by SSH analysis. Its expression levels in the hypothalamus and pituitary were significantly higher in brooding chickens than in non-brooding chickens. GWAS analysis with a NDH two tail sample showed that 2802 SNPs were significantly associated with egg number at 300 d of age (EN300). Among the 2802 SNPs, 2 SNPs composed a block overlapping the GARNL1 gene. The gene-centric GWAS analysis with another two tail sample of NDH showed that GARNL1 was strongly associated with EN300 and age at first egg (AFE). Single marker-trait association analysis in 1301 female NDH chickens confirmed that variation in this gene was related to EN300 and AFE. The allelic frequency spectrum of the SNP rs15700989 among 5 different populations supported the above associations. Western blotting, RT-PCR, and qPCR were used to analyze alternative splicing of the GARNL1 gene. RT-PCR detected 5 transcripts and revealed that the transcript, which has a 141 bp insertion, was expressed in a tissue-specific manner. CONCLUSIONS/SIGNIFICANCE: Our findings demonstrate that the GARNL1 gene contributes to chicken reproductive traits.