Exploring genotype-phenotype relationships of the CRABP2 gene on growth traits in beef cattle.

Cellular retinoic acid binding protein 2 (CRABP2) is essential to myoblast differentiation. However, there was no report about the function of CRABP2 gene in cattle. This study explored the association of CRABP2 gene polymorphisms with growth traits in cattle breeds by several methods, such as DNA sequencing, PCR, PCR-RFLP and forced PCR-RFLP. Two sequence variants were determined. There were 621 individuals in six cattle breeds from China for the experiment, and three breeds were used to test validation of polymorphisms and extent of linkage disequilibrium (LD). The results showed that both SNPs (SNP1, g.2458 G > T, SNP2, g.3878 G > A) were in intron1. Two SNPs were in low linkage disequilibrium. Association analysis suggested that SNP1 had the significant difference on growth traits with body height, height at hip cross and body slanting length (P < .05), while SNP2 showed a significant difference in growth traits with body height, height at hip cross and body slanting length(P < .05). The results of this investigation displayed that the CRABP2 gene is an available candidate gene and may be used for breed selection and conservation.

Integrating CNVs into meta-QTL identified GBP4 as positional candidate for adult cattle stature.

Copy number variation (CNV) of DNA sequences, functionally significant but yet fully ascertained, is believed to confer considerable increments in unexplained heritability of quantitative traits. Identification of phenotype-associated CNVs (paCNVs) therefore is a pressing need in CNV studies to speed up their exploitation in cattle breeding programs. Here, we provided a new avenue to achieve this goal that is to project the published CNV data onto meta-quantitative trait loci (meta-QTL) map which connects causal genes with phenotypes. Any CNVs overlapping meta-QTL therefore will be potential paCNVs. This study reported potential paCNVs in Bos taurus autosome 3 (BTA3). Notably, overview indexes and CNVs both highlighted a narrower region (BTA3 54,500,000-55,000,000 bp, named BTA3_INQTL_6) within one constructed meta-QTL. Then, we ascertained guanylate-binding protein 4 (GBP4) among the nine positional candidate genes was significantly associated with adult cattle stature, including body weight (BW, P < 0.05) and withers height (WHT, P < 0.05), fitting GBP4 CNV either with three levels or with six levels in the model. Although higher copy number downregulated the mRNA levels of GBP2 (P < 0.05) and GBP4 (P < 0.05) in 1-Mb window (54.0-55.0 Mb) in muscle and adipose, additional analyses will be needed to clarify the causality behind the ascertained association.

IL-4-Producing Dendritic Cells Induced during Schistosoma japonica Infection Promote Th2 Cells via IL-4-Dependent Pathway.

Although dendritic cells (DCs) have been widely demonstrated to play essential roles in initiation of Th2 responses in helminth infections and allergic reactions, the mechanisms remain uncertain largely because DCs do not produce IL-4. In present investigation, we have uncovered a novel subset of DCs from mice infected with Th2-provoking pathogens Schistosoma japonica, which independently promoted Th2 cells via IL-4-dependent pathway. These DCs contained similar levels of IL-4 mRNA and higher levels of IL-12p40 mRNA comparing to basophils, correlating to their Th2-promoting and Th1-promoting dual polarization capacities. Characterized by expression of FcεRI(+), these DCs were induced independent of T cells. Further investigations revealed that Th2-promoting FcεRI(+) DCs were monocyte-derived inflammatory DCs, which were sufficient to induce Th2 cells in vivo. Egg Ags together with GM-CSF or IL-3 alone were able to stimulate the generation of Th2-promoting FcεRI(+) DCs from bone marrow cells in vitro. To our knowledge, our data for the first time demonstrate that IL-4-producing DCs are induced under some Th2-provoking situations, and they should play important roles in initiation of Th2 response.

Interleukin-4- and NACHT, LRR and PYD domains-containing protein 3-independent mechanisms of alum enhanced T helper type 2 responses on basophils.

Aluminium hydroxide (alum), the most widely used adjuvant in human and animal vaccines, has long been known to promote T helper type 2 (Th2) responses and Th2-associated humoral responses, but the mechanisms have remained poorly understood. In this study, we explored whether alum is able to directly modulate antigen-presenting cells to enhance their potency for Th2 polarization. We found that alum treatment of dendritic cells failed to show any Th2-promoting activities. In contrast, alum was able to enhance the capacity of basophils to induce Th2 cells. When basophils from interleukin-4 (IL-4) knockout mice were examined, the intrinsic Th2-promoting activities by basophils were largely abrogated, but the alum-enhanced Th2-promoting activities on basophils were still detectable. More importantly, Th2-promoting adjuvant activities by alum found in IL-4 knockout mice were also largely reduced when basophils were depleted by antibody administration. Therefore, basophils can mediate Th2-promoting activities by alum both in vitro and in vivo through IL-4-independent mechanisms. Further studies revealed that secreted soluble molecules from alum-treated basophils were able to confer the Th2-promoting activities, and neutralization of thymic stromal lymphopoietin or IL-25 attenuated the IL-4-independent development of Th2 cells elicited by alum-treated basophils. Finally, alum was able to activate NACHT, LRR and PYD domains-containing protein 3 (NLRP3) inflammasome in murine basophils in the same way as alum in professional antigen-presenting cells, but NLRP3 was not required for Th2-promoting activities on basophils by alum in vitro. These results demonstrated that alum can enhance the capacities of basophils to polarize Th2 cells via IL-4- and NLRP3-independent pathways.

[Dynamic changes in professional and non-professional antigen presenting cells in the spleen from mice infected with Schistosoma japonicum].

UNLABELLED: OBJECTIVE; To observe the changes in various dendritic cell (DC) subsets, macrophages, basophils, eosinophils and mast cells in mouse spleen before and after Schistosoma japonicum infection induced Th2 response. METHODS: C57BL6 mice were infected with 20 S. japonicum cercariae via abdominal skin. Before infection and at 2, 4, and 6 weeks post-infection, the mice were sacrificed and spleen was removed. The frequencies of non-T, non-B basophils (NTNB), eosinophils, mast cells, subsets of DC and macrophages in the spleen were measured by flow cytometry. RESULTS: At 4 weeks after infection, when Th2 cells started to occur, the proportion of CD11c+CD8+DC and CD11c+CD4+DC in B220CD11c+DC increased from 7.4% and 7.9% before infection to 17.1% and 12.0%, respectively. During the infection, CD11c+CD4-CD8-DC, the majority of B220CD11c+DC, remained on a nearly constant level (70%); the percentage of B220+ CD11c+ DC in NTNB decreased. The macrophages were subdivided into two subsets: F4/80+CD11b(int) and F4/80+CD11b(high). The percentage of F4/80+CD11b(int) and F4/80+CD11b(high) in NTNB dropped from 15.4% and 13.7% before infection to 2.7% and 8.6% at 4 weeks post-infection. The proportion of CD11b high macrophages in F4/80+ cells significantly increased from 47.1% before infection to 75.5% at 4 weeks after infection. During S. japonicum infection, eosinophil percentage in the spleen gradually increased, while the frequency of basophils and mast cells in NTNB greatly decreased. CONCLUSION: At the time when Th2 response starts to occur, the frequency of CD11c+CD8+DC and CD11c+CD4+DC in CD11c+ dendritic cells increases. Once Th2 immune response established, the eosinophil frequency increases.

Sociodemographic Factors Associated With HIV/HCV High-Risk Behaviors Among People Who Use Drugs on Methadone Maintenance Treatment: A 10-Year Observational Study.

Background: Sociodemographic factors have an impact worldwide on the behavior of people who use drugs (PWUD). This study attempts to clarify the sociodemographic factors related to HIV/HCV high-risk behaviors (injection drug use, syringe sharing, and multiple sex partners) among PWUD on methadone maintenance treatment (MMT) in the long term. Methods: The 13,300 PWUD recruited into the MMT program were followed during 2006-2015. Generalized estimating equations were used to examine the relationship between sociodemographic characteristics and HIV/HCV high-risk behaviors. Results: We found that male (vs. female), living alone (vs. living with family or relatives), temporary income, financial support from family/friends, and financial support from social welfare (vs. regular salary) were positively associated with injection drug use. Age of initial drug use was negatively associated with injection drug use and syringe sharing. For both genders, being unmarried (vs. married or in cohabitation), living with friends, living alone (vs. living with family or relatives), temporary income, financial supports from family/friends (vs. regular salary), being employed (vs. unemployed/between jobs) was positively associated. In contrast, age at baseline was negatively associated with having multiple sexual partners for both genders. Ethnic of non-Han (vs. Han) was positively associated with having multiple sexual partners simply for males. Being divorced or widowed (vs. married or cohabitated) was positively associated with having multiple sexual partners merely for females. Conclusion: HIV/HCV high-risk behaviors correlated with certain sociodemographic factors of PWUD receiving MMT. There is a need for improving the well-being, employment, and housing status of PWUD on MMT to reduce their HIV/HCV risk behaviors.

Copy Number Variation of the SHE Gene in Sheep and Its Association with Economic Traits.

Copy number variation (CNV) caused by gene rearrangement is an important part of genomic structural variation. We found that the copy number variation region of the Src Homology 2 Domain Containing E (SHE) gene correlates with a quantitative trait locus of sheep related to milk fat percentage and bone density. The aim of our study was to detect the copy number variation of the SHE gene in four sheep breeds and to conduct a correlation analysis with economic traits, hoping to provide some reference for sheep breeding. In this study, we examined 750 sheep from four Chinese breeds: Chaka sheep (CKS), Hu sheep (HS), Large Tail Han sheep (LTHS) and Small Tail Han sheep (STHS). We used qPCR to evaluate the copy number of the SHE gene, and then used general linear models to analyze the associations between CNV and economic traits. The results showed that there were more individuals with SHE copy number loss in CKS and HS than in STHS and LTHS individuals. Association analyses showed that gain and normal copy number types were correlated to body length, circumference of cannon bone, heart girth, chest width and high at the cross in CKS, HS and STHS (p < 0.05), but this association was not observed for LTHS. Chi-square values (χ2) found prominent differences in CNV distribution among the studied breeds. Overall, the CNV of the SHE gene may be an important consideration for sheep molecular breeding.

Copy number variation (CNV) in the IGF1R gene across four cattle breeds and its association with economic traits.

The insulin-like growth factor 1 receptor (IGF1R) plays a vital role in immunomodulation and muscle and bone growth. The copy number variation (CNV) is believed to the reason for many complex phenotypic variations. In this paper, we statistically analyzed the copy number and the expression profiling in different tissue types of the IGF1R gene using the 422 samples from four Chinese beef cattle breeds, and the mRNA of IGF1R was widely expressed in nine tissue types of adult cattle (heart, liver, kidney, muscle, fat, stomach, spleen, lung and testis). Results of CNV and growth traits indicated that the IGF1R CNV was significantly associated with body weight and body height of Jinnan (JN) cattle and was significantly associated with body height and hucklebone width of Qinchuan (QC) cattle, making IGF1R CNV a promising molecular marker to improve meat production in beef cattle breeding. Bioinformatics predictions show that the CNV region is highly similar to the human genome, and there are a large number of transcription factors, DNase I hypersensitive sites, and high levels of histone acetylation, suggesting that this region may play a role in transcriptional regulation, providing directions for further study of the role of bovine CNV and economic traits.

Identification of Factors Influencing Out-of-county Hospitalizations in the New Cooperative Medical Scheme.

Throughout the duration of the New Cooperative Medical Scheme (NCMS), it was found that an increasing number of rural patients were seeking out-of-county medical treatment, which posed a great burden on the NCMS fund. Our study was conducted to examine the prevalence of out-of-county hospitalizations and its related factors, and to provide a scientific basis for follow-up health insurance policies. A total of 215 counties in central and western China from 2008 to 2016 were selected. The total out-of-county hospitalization rate in nine years was 16.95%, which increased from 12.37% in 2008 to 19.21% in 2016 with an average annual growth rate of 5.66%. Its related expenses and compensations were shown to increase each year, with those in the central region being higher than those in the western region. Stepwise logistic regression reveals that the increase in out-of-county hospitalization rate was associated with region (X1), rural population (X2), per capita per year net income (X3), per capita gross domestic product (GDP) (X4), per capita funding amount of NCMS (X5), compensation ratio of out-of-county hospitalization cost (X6), per time average in-county (X7) and out-of-county hospitalization cost (X8). According to Bayesian network (BN), the marginal probability of high out-of-county hospitalization rate was as high as 81.7%. Out-of-county hospitalizations were directly related to X8, X3, X4 and X6. The probability of high out-of-county hospitalization obtained based on hospitalization expenses factors, economy factors, regional characteristics and NCMS policy factors was 95.7%, 91.1%, 93.0% and 88.8%, respectively. And how these factors affect out-of-county hospitalization and their interrelationships were found out. Our findings suggest that more attention should be paid to the influence mechanism of these factors on out-of-county hospitalizations, and the increase of hospitalizations outside the county should be reasonably supervised and controlled and our results will be used to help guide the formulation of proper intervention policies.

Distribution and association study in copy number variation of KCNJ12 gene across four Chinese cattle populations.

Copy number variation is a large genome variation which usually happens in the noncoding-region, and it may occur at the locus associated with the functional gene to further influence the phenotype. Potassium inwardly-rectifying channel, subfamily J 12 (KCNJ12) gene expressed widely in cardiomyocytes and neurons, plays an important role in tumor therapy and muscle movement regulation. In this study, we detected the distribution of CNVs for KCNJ12 gene in 404 individuals belonging to four Chinese cattle breeds (NY, JX, JA and GF). We also investigated the KCNJ12 gene expression in different tissues of JX cattle. Additionally, we examined the association of two CNV regions (CNV1: 1,600 bp, intron 1; CNV2: 4,800 bp, intergenic) with growth traits. The statistical analyses indicated that the CNV1 is associated with the body length, rump length and weight in JX cattle population (P < 0.05); and there has a significant association with the body length, chest circumference, and body weight in GF cattle (P < 0.05).The CNV2 had a significant effect on the body length and body weight in JX cattle (P < 0.05); the body length, chest circumference, rump length and body weight in GF cattle (P < 0.01 or P < 0.05). The copy numbers of KCNJ12 gene presented the negative correlations with the transcript level of gene in skeletal muscles (P < 0.05). Our results provide evidence that CNV1 and CNV 2 in KCNJ12 are associated with growth traits in two cattle populations and may be used as candidates for marker-assisted selection and breeding management in cattle.