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Objective: To observe the clinical efficacy of modified painless wheat-grain blistering moxibustion for allergic rhinitis (AR) of lung deficiency and cold attacking, and to explore its effects on serum immunoglobulin E (IgE) and interleukin-10 (IL-10)., Methods: Ninety-eight patients of perennial AR with lung deficiency and cold attacking were randomly divided into an observation group (49 cases, 2 dropped out) and a control group (49 cases, 2 dropped out). The control group received mometasone furoate nasal spray treatment. The observation group received modified painless wheat-grain blistering moxibustion at bilateral Feishu (BL 13), Gaohuang (BL 43), Zusanli (ST 36), and Shenzhu (GV 12) in addition to the control group's treatment. Moxibustion at Shenzhu (GV 12) was applied once every other day, 3 grains each time, forming moxibustion sores after about one week. After sores formed, moxibustion was applied once every other 2 days. For Feishu (BL 13), Gaohuang (BL 43), and Zusanli (ST 36), moxibustion was applied on one side first, every other day, 3 grains each time, until sores formed, then on the other side, alternating sides in a cycle. Both groups were treated for 4 weeks. The total nasal symptoms score (TNSS), nasal symptom visual analogue scale (VAS), and rhinoconjunctivitis quality of life questionnaire (RQLQ) scores were observed before and after treatment, and after 4 and 12 weeks of treatment completion (follow-ups). Serum IgE and IL-10 levels were measured before and after treatment, and treatment efficacy and recurrence rates at follow-ups were recorded., Results: Compared before treatment, TNSS, VAS, and RQLQ scores in both groups were reduced after treatment and at follow-ups ( P <0.05), and these scores in the observation group were lower than those in the control group ( P <0.05). Except for TNSS scores in the control group at the follow-ups, and in the observation group at the 4-week follow-up, all scores at follow-ups in both groups were higher than those after treatment ( P <0.05). Compared before treatment, serum IgE levels in both groups were decreased ( P <0.05), and serum IL-10 levels were increased ( P <0.05) after treatment. The observation group had lower serum IgE levels and higher IL-10 levels than the control group ( P <0.05). The total effective rate in the observation group was 93.6% (44/47), higher than 74.5% (35/47) in the control group ( P <0.05). The recurrence rates after 4 and 12 weeks of treatment completion in the observation group were lower than those in the control group (4.5% [2/44] vs 22.9% [8/35], 9.1% [4/44] vs 40.0% [14/35], P <0.05)., Conclusion: On the basis of mometasone furoate nasal spray, modified painless wheat-grain blistering moxibustion could improve clinical symptoms in patients of AR with lung deficiency and cold attacking, and provide more sustained long-term efficacy, possibly through the regulation of serum IgE and IL-10 levels.

Buffalo colostrum is the initial mammary secretion after parturition, consisting of nutritional and bioactive components. In this study, we conducted a proteomic analysis of buffalo colostrum whey to identify bioactive proteins and peptides. A total of 107 differentially expressed proteins (DEPs) were identified in buffalo colostrum whey compared to those in mature milk. Gene Ontology analysis revealed that DEPs were primarily associated with immune response and tissue development. KEGG pathway enrichment suggested that colostrum actively enhances nascent immunity involved in interleukin and interferon signaling pathways. Furthermore, candidate antimicrobial peptides (AMPs) of whey protein hydrolysates from buffalo colostrum were characterized, which exhibits broad-spectrum activity against gram-positive and gram-negative pathogens. Overall, this study improves our understanding of protein variations in buffalo lactation, and contributes to the development of AMPs from buffalo colostrum., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Spermatogenesis carries the task of precise intergenerational transmission of genetic information from the paternal genome and involves complex developmental processes regulated by the testicular microenvironment. Studies performed mainly in mouse models have established the theoretical basis for spermatogenesis, yet the wide interspecies differences preclude direct translation of the findings, and farm animal studies are progressing slowly. More than 32,000 cells from prepubertal (3-month-old) and pubertal (24-month-old) buffalo testes were analyzed by using single-cell RNA sequencing (scRNA-seq), and dynamic gene expression roadmaps of germ and somatic cell development were generated. In addition to identifying the dynamic processes of sequential cell fate transitions, the global cell-cell communication essential to maintain regular spermatogenesis in the buffalo testicular microenvironment was uncovered. The findings provide the theoretical basis for establishing buffalo germline stem cells in vitro or culturing organoids and facilitating the expansion of superior livestock breeding., Competing Interests: The authors declare no competing interests., (© 2022 The Author(s).)

Phthalates (PAEs) are common endocrine disrupting chemicals (EDCs) that disrupt fetal development. The present study aimed to evaluate the effects of single and coexposure to phthalates in early pregnancy on fetal growth restriction (FGR) by a nested case-control study based on the Guangxi Zhuang Birth Cohort (GZBC). Maternal serum concentrations of seven phthalates in 97 neonates with FGR and 291 matched controls were detected through gas chromatography-mass spectrometry (GC-MS). The associations between phthalates and FGR were analyzed using multiple logistic regression, weight quantile sum (WQS) regression, and Bayesian kernel machine regression (BKMR) models. We found that exposures to butyl-benzyl phthalate (BBP, OR adj  = 1.849, 95% CI: 1.080-3.177, P adj  = 0.025, P trend  = 0.046), di (2-ethyl-hexyl) phthalate (DEHP, OR adj  = 3.893, 95% CI: 1.305-11.910, P adj  = 0.015, P trend  = 0.098) and dimethyl phthalate (DMP, OR adj  = 1.722, 95% CI: 1.089-2.725, P adj  = 0.020, P trend  = 0.002) were significantly positively associated with the risk of FGR, while mono-butyl phthalate (MBP) showed a significant negative association with FGR (OR high  = 0.192, 95% CI: 0.036-0.795, P adj  = 0.033, P trend  = 0.035) only among girls. The WQS model identified that BBP, di(2-ethyl)phthalate (DEP), DMP, DEHP, di-n-butyl phthalate (DBP), and MBP were highly weighted in the association with FGR. The BKMR model supported the positive association between joint exposure to phthalates and the risk of FGR and identified no significant interaction between the seven phthalates. Overall, maternal exposure to BBP, DEHP, and DMP may cause adverse effects on FGR, especially with combined effects., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The acquisition of mammalian sperm motility is a main indicator of epididymal sperm maturation and helps ensure fertilization. Poor sperm motility will prevent sperm cells from reaching the fertilization site, resulting in fertilization failure. To investigate the proteomic profiling of normal and poorly motile buffalo spermatozoa, a strategy applying liquid chromatography tandem mass spectrometry combined with tandem mass targeting was used. As a result, 145 differentially expressed proteins (DEPs) were identified in poorly motile spermatozoa (fold change > 1.5), including 52 upregulated and 93 downregulated proteins. The upregulated DEPs were mainly involved in morphogenesis and regulation of cell differentiation. The downregulated DEPs were involved with transport, oxidation-reduction, sperm motility, regulation of cAMP metabolism and regulation of DNA methylation. The mRNA and protein levels of PRM1 and AKAP3 were lower in poorly motile spermatozoa, while the expressions of SDC2, TEKT3 and IDH1 were not correlated with motility, indicating that their protein changes were affected by transcription or translation. Such changes in the expression of these proteins suggest that the formation of poorly motile buffalo spermatozoa reflects a low efficiency of energy metabolism, decreases in sperm protamine proteins, deficiencies in motility-related proteins, and variations in tail structural proteins. Such proteins could be biomarkers of poorly motile spermatozoa. These results illustrate some of the molecular mechanisms associated with poorly motile spermatozoa and provide clues for finding molecular markers of these pathways.

Alkylphenols are a type of endocrine disruptors, which are commonly found in personal care products, food, and water and are more harmful to the human body. To investigate the relationship between exposure of alkylphenols in serum of pregnant women during early pregnancy and adverse birth outcomes, a total of 2035 healthy pregnant women and their neonates were recruited in the birth cohort of Zhuang nationality in Guangxi from 2015 to 2018. The peripheral venous blood samples were collected from pregnant women in early pregnancy; the concentrations of nonylphenol (NP), 4-nonylphenol (4-N-NP), 4-tert-octylphenol (4-T-OP), and 4-n-octylphenol (4-N-OP) in serum were detected by ultra-performance liquid performance chromatography-tandem mass spectrometry (UPLC-MS). Binary logistic regression showed that NP [OR = 1.40 (95% CI: 1.00, 1.94)] was positively associated with preterm birth. Restricted cubic spline analyses showed that logNP and log4-T-OP had non-linearity associations with preterm birth (logNP: P overall = 0.006, P non-linear = 0.003; log4-T-OP: P overall = 0.004, P non-linear = 0.002). Multiple linear regression analysis showed that maternal serum concentration of NP was negatively associated with birth weight of perinatal infants (β = -14.68, 95% CI: -29.18, -0.19), which may be sensitive in male neonates (β = -26.18, 95% CI: -47.33, -5.02). The findings demonstrate that nonylphenol is a risk factor of preterm birth, and nonylphenol is negatively associated with the birth weight in male infants., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Maternal-effect genes (MEGs) accumulate in oocytes during oogenesis and mediate the pre-implantation embryo developmental programme until activation of the zygote genome. Nlrp5 and Tle6 are required for normal pre-implantation and embryonic development. However, the precise function of these MEGs in buffalo (Bubalus bubalis) remains to be elucidated. The aim of this study was to characterize Nlrp5 and Tle6 sequences and analyse their mRNA and protein expression patterns in somatic tissues, oocytes and pre-implantation embryos of buffalo. The coding sequences of each gene were successfully cloned and characterized. Real-time quantitative reverse transcription PCR results revealed an absence of Nlrp5 or Tle6 transcripts in somatic tissues, with the exception of ovary. Expression levels of Nlrp5 and Tle6 in oocytes increased from the germinal vesicle stage to metaphase II stage and then gradually decreased during morula and blastocyst stages. Protein expression patterns were confirmed by immunofluorescence analysis. This study lays a foundation for further validation of the function of MEGs in buffalo., (© 2022 Wiley-VCH GmbH.)

Endoplasmic reticulum (ER) stress plays a crucial role in granulosa cell (GCs) apoptosis, which is the main cause of follicular atresia. Quercetin (QC), a plant-derived flavonoid, has antioxidant, anti-inflammatory, and other biological properties. However, whether QC can alleviate the effects of ER stress on buffalo GCs remains unknown. In this study, we constructed an ER stress model in buffalo GCs by using tunicamycin (TM) and pre-treated with QC to explore the effect of QC on cells under ER stress. Apoptosis was detected by Annexin fluorescein 5 isothiocyanate (V-FITC), and the expressions of mRNA and related proteins involved in ER stress and apoptosis were detected via real-time polymerase chain reaction and Western blot. The results revealed that ER stress can cause apoptosis in GCs, whereas QC pre-treatment can prevent apoptosis caused by ER stress. After pre-treatment with QC, the expression levels of ER stress-related genes and proteins significantly decreased, pro-apoptotic genes were significantly down-regulated, and anti-apoptotic genes were significantly up-regulated. Furthermore, the results of Chop gene overexpression suggested that QC alleviated ER stress via the PERK/CHOP signaling pathway. In this study, we preliminarily elucidated that QC alleviates ER stress-induced apoptosis in buffalo GCs, and the results suggest a novel strategy for delaying follicular atresia by inhibiting GCs apoptosis.

Bromodomain (BRD) is an evolutionarily conserved protein-protein interaction module that is critical in gene regulation, cellular homeostasis, and epigenetics. This study aimed to conduct an identification, evolution, and expression analysis of the BRD gene family in the swamp buffalo ( Bubalus bubalis ). A total of 101 BRD protein sequences deduced from 22 BRD genes were found in the buffalo genome. The BRD proteins were classified into six groups based on phylogenetic relationships, conserved motifs, and conserved domains. The BRD genes were irregularly distributed in 13 chromosomes. Collinearity analysis revealed 20 BRD gene pairs that had remarkable homologous relationships between the buffalo and cattle, although no tandem or segmental duplication event was found in the buffalo BRD genes. Comparative transcriptomics using a 10x sequencing platform analysis showed that 22 BRD genes were identified in the Sertoli cells (SCs) at different developmental stages of buffalo. Further, the mRNA expression levels of bromodomain and the extraterminal (BET) family in SCs at the pubertal stage were higher than that at the prepubertal stage of buffalo. However, the SMARCA2 , PHIP , BRD9 , and TAF1 genes exhibited the opposite trend. The maturation process of SCs may be regulated by the BRD family members expressed differentially in SCs at different developmental stages of buffalo. In summary, our findings provide an understanding of the evolutionary, structural, and functional properties of the buffalo BRD family members, and further characterize the function of the BRD family in the maturation of SCs. It also provides a theoretical basis for further understanding in the future of the mechanism of SCs regulating spermatogenesis.

Leydig cells (LCs) are testosterone-generating endocrine cells that are located outside the seminiferous tubules in the testis, and testosterone is fundamental for retaining spermatogenesis and male fertility. In buffalo, adult Leydig cells (ALCs) are developed by immature Leydig cells (ILCs) in the postnatal testes. However, the genes/pathways associated to the regulation of testosterone secretion function during the development of postnatal LCs remains comprehensively unidentified. The present study comparatively analyzed the transcriptome profiles of ILC and ALC in buffalo with significant differences in testosterone secretion. Differentially expressed genes (DEGs) analysis identified 972 and 1,091 annotated genes that were significantly up- and down-regulated in buffalo ALC. Functional enrichment analysis showed that cAMP signaling being the most significantly enriched pathway, and testosterone synthesis and lipid transport-related genes/pathways were upregulated in ALC. Furthermore, gene set enrichment analysis (GSEA) shows that cAMP signaling and steroid hormone biosynthesis were activated in ALC, demonstrating that cAMP signaling may serve as a positive regulatory pathway in the maintenance of testosterone function during postnatal development of LCs. Protein-protein interaction (PPI) networks analysis highlighted that ADCY8, ADCY2, POMC, CHRM2, SST, PTGER3, SSTR2, SSTR1, NPY1R, and HTR1D as hub genes in the cAMP signaling pathway. In conclusion, this study identified key genes and pathways associated in the regulation of testosterone secretion function during the ILC-ALC transition in buffalo based on bioinformatics analysis, and these key genes might be deeply involved in cAMP generation to influencing testosterone levels in LCs. The results suggest that ALCs might increase testosterone levels by enhancing cAMP production than ILCs. Our data will enhance the understanding of developmental mechanism studies related to testosterone function and provide preliminary evidence for molecular mechanisms of LCs regulating spermatogenesis., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Sertoli cells provide nutrients and support for germ cell differentiation and maintain a stable microenvironment for spermatogenesis. Comprehensive identification of Sertoli cellular proteins is important in understanding spermatogenesis. In this study, we performed an integrative analysis of the proteome and phosphoproteome to explore the role of Sertoli cells in spermatogenesis. A total of 2912 and 753 proteins were identified from the proteome and phosphoproteome in Sertoli cells, respectively; 438 proteins were common to the proteome and phosphoproteome. Data are available via ProteomeXchange with identifier PXD024984. In the proteome, ACTG1, ACTB, ACTA2, MYH9 were the most abundant proteins. Gene Ontology (GO) analysis indicated that most of the proteins were involved in the processes of localization, biosynthesis, gene expression, and transport. In addition, some of the proteins related to Sertoli cell functions were also enriched. In the phosphoproteome, most of the proteins were involved in gene expression and the RNA metabolic process; the pathways mainly involved the spliceosome, mitogen-activated protein kinase signaling pathway, focal adhesion, and tight junctions. The pleckstrin homology-like domain is the most highly enriched protein domain in phosphoproteins. Cyclin-dependent kinases and protein kinases C were found to be highly active kinases in the kinase-substrate network analysis. Ten proteins most closely related to network stability were found in the analysis of the network interactions of proteins identified jointly in the phosphoproteome and proteome. Through immunohistochemistry and immunofluorescence verification of vimentin, it was found that there were localization differences between phosphorylated and non-phosphorylated vimentin in testicular tissue. This study is the first in-depth proteomic and phosphoproteomic analysis of buffalo testicular Sertoli cells. The results provide insight into the role of Sertoli cells in spermatogenesis and provide clues for further study of male reproduction., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2021 Elsevier Inc. All rights reserved.)

In this paper, we present a simultaneous and continuous kinematics estimation method for multiple DoFs across shoulder and elbow joint. Although simultaneous and continuous kinematics estimation from surface electromyography (EMG) is a feasible way to achieve natural and intuitive human-machine interaction, few works investigated multi-DoF estimation across the significant joints of upper limb, shoulder and elbow joints. This paper evaluates the feasibility to estimate 4-DoF kinematics at shoulder and elbow during coordinated arm movements. Considering the potential applications of this method in exoskeleton, prosthetics and other arm rehabilitation techniques, the estimation performance is presented with different muscle activity decomposition and learning strategies. Principle component analysis (PCA) and independent component analysis (ICA) are respectively employed for EMG mode decomposition with artificial neural network (ANN) for learning the electromechanical association. Four joint angles across shoulder and elbow are simultaneously and continuously estimated from EMG in four coordinated arm movements. By using ICA (PCA) and single ANN, the average estimation accuracy 91.12% (90.23%) is obtained in 70-s intra-cross validation and 87.00% (86.30%) is obtained in 2-min inter-cross validation. This result suggests it is feasible and effective to use ICA (PCA) with single ANN for multi-joint kinematics estimation in variant application conditions.