Investigation into the association of FNDC1 and ADAMTS12 gene expression with plumage coloration in Muscovy ducks.

To elucidate the molecular genetic mechanisms underpinning feather color in Muscovy ducks. A cohort of 100 Muscovy ducks was meticulously selected for this research. Follicular tissues from ducks exhibiting black and white plumage served as the experimental samples. From these tissues, RNA and proteins were extracted for further analysis. The RNA underwent reverse transcription polymerase chain reaction amplification, followed by validation through western blot assays. The data revealed a significant upregulation in the expression of FN domain-containing protein 1 (FNDC1) and ADAMTS12 genes in Muscovy ducks with white plumage traits as opposed to those with black plumage traits. Specifically, individuals with pure white plumage demonstrated a markedly elevated expression of the FNDC1 gene in comparison to their pure black counterparts. Conversely, expression levels of the ADAMTS12 gene were found to be reduced in ducks with pure black plumage relative to those with pure white plumage. Notably, the expression patterns of FNDC1 and ADAMTS12 genes exhibited inconsistencies between mRNA and protein levels. This study offers significant insights into the molecular genetic mechanisms underlying feather color variation in Muscovy ducks. FNDC1 and ADAMTS12 could be considered potential targets for genetic manipulation or selective breeding strategies aimed at achieving specific feather color phenotypes in Muscovy ducks.

Exploration of the genetic influence of MYOT and MB genes on the plumage coloration of Muscovy ducks.

Plumage color, a pivotal attribute delineating diverse Muscovy duck strains, assumes considerable significance within the field of Muscovy duck breeding research. This study extends the existing research by delving into the hereditary aspects of genes associated with plumage coloration in Muscovy ducks. The principal objective is to discern marker genes conducive to targeted breeding strategies based on plumage color, thereby furnishing indispensable technical foundations for the development of novel Muscovy duck varieties. Our investigation focused on scrutinizing the impact of MYOT and MB genes on the genetic expression of plumage color at both the RNA and protein levels in Muscovy ducks. The results elucidate that black Muscovy ducks manifest markedly elevated mRNA and protein expression levels of MYOT and MB genes in comparison to their white counterparts, indicating that both genes may play a constructive regulatory role in the context of plumage coloration in Muscovy ducks. The outcomes of this study delineate a discernible correlation between MYOT and MB genes and the plumage coloration in Muscovy ducks. Employing gene expression analysis, we successfully identified candidate genes that may be intricately linked to the determination of plumage color in these ducks.

Multiple stimulus-response berberine plus baicalin micelles with particle size-charge-release triple variable properties for breast cancer therapy.

OBJECTIVE: The aim was to develop a nanoscale drug delivery system with enzyme responsive and acid sensitive particle size and intelligent degradation aiming to research the inhibitory effect on breast cancer. SIGNIFICANCE: The delivery system addressed the problems of tissue targeting, cellular internalization, and slow drug release at the target site, which could improve the efficiency of drug delivery and provide a feasible therapeutic approach for breast cancer. METHODS: The acid sensitive functional material DSPE-PEG2000-dyn-PEG-R9 was synthesized by Michael addition reaction. Then, the berberine plus baicalin intelligent micelles were prepared by thin-film hydration. Subsequently, we characterized the physical and chemical properties of berberine plus baicalin intelligent micelles, evaluated its anti-tumor effects in vivo and in vitro. RESULTS: The target molecule was successfully synthesized, and the intelligent micelles showed excellent chemical and physical properties, delayed drug release and high encapsulation efficiency. In vitro and in vivo experiments also confirmed that the intelligent micelles could effectively target tumor sites, penetrate tumor tissues, enrich in tumor cells, inhibit tumor cell proliferation, inhibit tumor cell invasion and migration, and induce tumor cell apoptosis. CONCLUSION: Berberine plus baicalin intelligent micelles have excellent anti-tumor effects and no toxicity to normal tissues, which provides a new potential drug delivery strategy for the treatment of breast cancer.

Effect of resveratrol treatment on apoptosis and apoptotic pathways during boar semen freezing.

Resveratrol (3,5,4'-trihydroxystilbene, RSV) has been widely used in mammalian cells, but whether it can be used during freezing boar semen is still unknown. The effects of RSV treatment during boar semen freezing on its anti-freezing ability, apoptosis, and possible apoptotic pathways were observed in this study. Sperm motility, mitochondrial membrane potential (ΔΨm), adenosine triphosphate (ATP) content, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labeling (TUNEL)-positive apoptotic state, and messenger RNA (mRNA) expression levels of apoptotic genes involved in different apoptotic pathways after freezing with or without RSV treatment were tested. The results showed that: (1) Compared with fresh sperm, the motility, normal acrosome rate, and plasma membrane integrity rate of frozen boar sperm decreased significantly (P<0.05), and RSV did not significantly increase the sperm motility (0.44 vs. 0.40, P>0.05), but it did significantly improve the normal acrosome rate (57.65% vs. 47.00%, P<0.05) and plasma membrane integrity rate (46.67% vs. 38.85%, P<0.05). (2) After freezing, most boar sperm showed low mitochondrial ΔΨm. RSV treatment could increase the rate of high mitochondrial ΔΨm of boar sperm. (3) RSV treatment significantly decreased reactive oxygen species (ROS) levels (58.65% vs. 88.41%, P<0.05) and increased the ATP content (0.49 µmol/L vs. 0.25 µmol/L, P<0.05) of boar sperm during freezing. (4) The apoptotic rate of the freezing group (80.41%) with TUNEL detection increased significantly compared to the fresh group (9.70%, P<0.05), and RSV treatment greatly decreased the apoptotic rate (68.32%, P<0.05). (5) Real-time polymerase chain reaction (RT-PCR) showed that not only the genes from the death receptor-mediated apoptotic pathway (tumor necrosis factor-α (TNF-α), Fas ligand (FasL), and Caspase-8), but also the genes from the mitochondria-mediated apoptotic pathway (manganese superoxide dismutase (MnSOD), B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), and Caspase-9) were both significantly changed after freezing. RSV treatment during freezing greatly changed their expression levels. Although RSV treatment during boar semen freezing did not significantly increase motility after thawing, it still played an efficient antioxidant role, which could enhance the mitochondrial function and decrease the apoptotic level induced by both the death receptor- and mitochondria-mediated apoptotic pathways.

The enhanced treatment efficacy of invasive brain glioma by dual-targeted artemether plus paclitaxel micelles.

High grade-gliomas are highly invasive and prone to metastasis, leading to poor survival and prognosis. Currently, we urgently need a new treatment strategy to effectively inhibit glioma. In this study, artemether and paclitaxel were used as two agents for tumour suppression. Two functional materials were synthesised and modified on the surface of the micelle as targeting molecules. The addition of two functional materials confers the ability of the micelles to effectively cross the blood-brain barrier (BBB) and then target the glioma cells. Thus, this dual-targeted delivery system allows the drug to play a better role in inhibiting tumour invasion and vasculogenic mimicry (VM) channels. In this paper, the anticancer effects of dual-targeted artemether plus paclitaxel micelles on glioma U87 cells were studied in three aspects: (I) In vitro and in vivo targeting assessment, including the role of penetrating BBB and targeting glioma; (II) In vitro regulation of invasion-associated proteins; (III) Inhibition of VM channels formation and invasion in vitro; (IV) The study of pharmacodynamics in tumour-bearing mice. These results suggest that dual-targeted artemether plus paclitaxel micelle may provide a new strategy to treat glioma via inhibiting invasive and VM channels.

Understanding the cross-talk between host and virus in poultry from the perspectives of microRNA.

In poultry, viral infections (e.g., Marek's disease virus, avian leukosis virus, influenza A virus, and so on) can cause devastating mortality and economic losses. Because viruses are solely dependent on host cells to propagate, they alter the host intracellular microenvironment. Thus, understanding the virus-host interaction is important for antiviral immunity and drug development in the poultry industry. MicroRNAs are crucial posttranscriptional regulators of gene expression in a wide spectrum of biological processes, including viral infection. Recently, microRNAs have been identified as key players in virus-host interactions. In this review, we will discuss the intricacies involved in the virus-host cross-talk mediated by host and viral microRNAs in poultry (i.e., chicken and ducks), as well as recent trends and challenges in this field. These findings may provide some insights into the rapidly developing area of research regarding viral pathogenesis and antiviral immunity in poultry production.

Transcriptomic analysis of different stages of pigeon ovaries by RNA-sequencing.

The pigeon ovary is an ideal model for deciphering the molecular mechanism of folliculogenesis. While most analysis has focused on the influence of hormones and factors on ovarian follicle development in this model, changes occurring in the ovarian stroma can also be extremely informative. Here, we profiled the transcriptome of pigeon ovaries at pre-ovulation, post-ovulation, and 5-6 days after ovulation using RNA-sequencing to gain insights into the molecular and cellular events mediating ovary activity. We obtained 44,784,505 clean reads that aligned with 14,088 genes. Gene expression profile analysis identified 409 differentially expressed genes between pre- and post-ovulation; 96 genes were up-regulated genes while 313 genes were down-regulated. Gene ontology analysis of the down-regulated genes revealed significant enrichment in components of the immune response, immune system, antigen processing and presentation, receptor binding, and biological adhesion. Pathway analyses of the high-expression genes of the post-ovulation ovary identified enrichment in phagosomes, lysosomes, cytokine-cytokine receptor interactions, cell adhesion molecules, and the Toll-like receptor signaling. These data together suggest that post-ovulatory follicle regression and elimination occurs through an immune response. Mol. Reprod. Dev. 83: 640-648, 2016. © 2016 Wiley Periodicals, Inc.

Nuclear effects of G-protein receptor kinase 5 on histone deacetylase 5-regulated gene transcription in heart failure.

BACKGROUND: G-protein receptor kinases (GRKs) modulate cardiac ß-adrenergic signaling. GRK5 is upregulated in heart failure, and a gain-of-function polymorphism substituting leucine for wild-type glutamine at amino acid 41 (GRK5-Leu41) is associated with improved outcomes in heart failure and hypertension. GRK5 is distinguished by partial nuclear localization and class II histone deacetylases (HDAC) kinase activity that is postulated to regulate Gαq-stimulated cardiac gene expression. METHODS AND RESULTS: We used in vitro tissue culture and in vivo mouse compound genetic models to examine the effects of GRK5 on HDAC phosphorylation, nucleo-cytoplasmic HDAC transport, and Gαq-dependent transcriptional regulation. In vitro, GRK5 stimulated HDAC5 nuclear export only in the context of Gαq signaling stimulated by angiotensin II. GRK5-Gln41 and Leu41 were similar inducers of HDAC5 nucleo-cytoplasmic shuttling. In vivo, GRK5-Gln41 and-Leu41 partitioned equally to nuclear and nonnuclear myocardial fractions. GRK5 increased cardiac HDAC5 phosphorylation and reversed the increase in nuclear HDAC5 content seen with cardiomyocyte-autonomous Gαq overexpression. Deep RNA sequencing showed few changes in gene expression induced by GRK5 overexpression or ablation alone, but GRK5 overexpression normalized steady-state expression levels of 48% (96 of 200) of all Gαq down-regulated mRNAs. CONCLUSIONS: GRK5 is a transcriptional modifier of a subset of Gαq-downregulated genes, acting in opposition to the pathological effects of Gαq and normalizing levels of these transcripts. This transcriptional coregulator effect may act in concert with ß-adrenergic receptor desensitization to protect against heart failure decompensation.

Receptor-independent protein kinase C alpha (PKCalpha) signaling by calpain-generated free catalytic domains induces HDAC5 nuclear export and regulates cardiac transcription.

Receptor-mediated activation of protein kinase (PK) C is a central pathway regulating cell growth, homeostasis, and programmed death. Recently, we showed that calpain-mediated proteolytic processing of PKCα in ischemic myocardium activates PKC signaling in a receptor-independent manner by releasing a persistent and constitutively active free catalytic fragment, PKCα-CT. This unregulated kinase provokes cardiomyopathy, but the mechanisms remain unclear. Here, we demonstrate that PKCα-CT is a potent regulator of pathological cardiac gene expression. PKCα-CT constitutively localizes to nuclei and directly promotes nucleo-cytoplasmic shuttling of HDAC5, inducing expression of apoptosis and other deleterious genes. Whereas PKD activation is required for HDAC5 nuclear export induced by unprocessed PKCs activated by phorbol ester, PKCα-CT directly drives HDAC cytosolic relocalization. Activation of MEF2-dependent inflammatory pathway genes by PKCα-CT can induce a cell-autonomous transcriptional response that mimics, but anticipates, actual inflammation. Because calpain-mediated processing of PKC isoforms occurs in many tissues wherein calcium is increased by stress or injury, our observation that the catalytically active product of this interaction is a constitutively active transcriptional regulator has broad ramifications for understanding and preventing the pathological transcriptional stress response.

[Compatibility chemistry of acid-alkaline pair medicine of Dahuang and Huangbai in Dahuang Xiaoshi decoction].

OBJECTIVE: To determine the change pattern of alkaline and acid components of Dahuang and Huangbai amoung different combinations in Dahuang Xiaoshi decotion. METHOD: The contents of anthraquinones and berberine were determined by HPLC in samples of Dahuang extracts, Huangbai extracts, Dahuang and Huangbai pair medicine extracts, and extracts of whole recipe. RESULT: The contents of anthraquinones and berberine were decreased with the changes of combinations: from component medicinal material alone to pair medicines, and to whole recipe. the change pattern was more apparent in water decoctions than in ethanol extracts. CONCLUSION: The contents of alkaline and acid components are changed with different extract methods and different combinations.

[Compatability chemistry of acid-alkaline pair medicine of dahuang and Fuzi in Dahuang Fuzi decoction].

OBJECTIVE: To determine the change pattern of alkaline and acid components of Dahuang and Fuzi in different combinations. METHOD: The contents of anthraquinones and aconite alkaloids were determined by HPLC in samples of Dahuang extracts, Fuzi extracts, Dahuang and Fuzi pair medicines extracts, and extracts of whole recipe. RESULT: As for Dahuang, the contents of anthraquinones were decreased with changes of combination: higher contents were in decoctions of Dahuang alone, lower contents were in decoctions of pair medicines, and the lowest contents were in decoctions of whole recipe. As for Fuzi, monoester aconite alkaloids could be detected in water extracts, and both biester and monoester aconite alkaloids could be detected in ethanol extracts. The contents of aconite alkaloids were decreased with changes of combinations: from Fuzi alone to pair medicines, and to whole recipe. The change pattern was more apparent when extracted in water decoction. CONCLUSION: The contents of alkaline and acid components are changed with different extract methods and different combinations.

[Compatability chemistry of acid-alkaline pair medicine of Fuzi and Gancao in Sini decoction].

OBJECTIVE: To determine the change pattern of alkaline and acid components of Fuzi and gancao amoung different combinations in Sini Decotion. METHOD: The contents of aconite alkaloids and glycyrrhizic acid were determined by HPLC in samples of Fuzi extracts, gancao extracts, Fuzi and gancao pair medicines extracts, and extracts of whole recipe. RESULT: As for Fuzi, monoester aconite alkaloids could be detected in water extracts, and both biester and monoester aconite alkaloids could be detected in ethanol extracts. The contents of aconite alkaloids were decreased with changes of combinations: from Fuzi alone to pair medicines, and to whole recipe. The change pattern was more apparent when extracted in water decoction. As for Gancao, the glycyrrhizic acid was lowered from single medicine to pair medicines, and to whole recipe. CONCLUSION: The contents of alkaline and acid components were changed with different extract methods and different combinations of Fuzi and Gancao.

[Research on compatibility chemistry of acid-alkaline pair medicines in formulas of traditional Chinese medicine].

Compatibility chemistry of acid-alkaline pair medicines in formulas of traditional Chinese medicine (TCM) is an important research field which should merit to pay attention. The ideas and methods in prescription compatibility research on formulas containing alkaline-acid pair medicines were summarized from the aspect of chemical groups of alkaline and acid ingredients; the research results were introduced and analyzed; the research meaning was elaborated; and the expectation of the field was viewed.

[Compatability chemistry of acid-alkaline pair medicine of Ephedra sinica and Glycyrrhiza uralencis in Maxing Shigan decoction].

OBJECTIVE: To investigate the change regularity of ephedrine and glyrihhzine acid in Ephedra sinila and Glycyrrhiza uralencis pair medicines and in Maxing Shigan decoction. METHOD: The contents of ephedrine and glycyrrhizic acid were determined by HPLC in samples of E. sinica extracts, G. uralencis extracts, pair medicines extracts of Maxing Shigao decoction sinica and G. uralencis, and extracts of E. sinica. RESULT: There were no significant difference in ephedrine contents amoung different samples; the contents of glycyrrhizic acid were lower in decoctions of Maxing Shigan decoction than in G. uralencis decoction and pair medicines extracts. CONCLUSION: Macromolecular complex was NOT formed by ephedrine and Glycyrrhizic acid in decoctions containing pair medicines of E. sinica and G. uralencis.

[Studies on chemical structure of complexes compounds formed by acid and alkaline components of pair medicines in decoction].

OBJECTIVE: To study the chemical structures of macromolecular complexes formed by aconitine and glycyrrhizic acid, aconitine and rhein, berberine and rhein, ephedrine and glycyrrhizic acid in decoctions. METHOD: The binding energy of reactants and products was determined by X-ray spectrography, the ground level charge distribution of products was caculated by density function theory of quantum chemistry. RESULT: The binding energy of aconitine and glycyrrhizic acid, aconitine and rhein, berberine and rhein were changed after reaction. The characteristic atoms in the molecular structures of acid components have higher positive electric charge, while the ones in alkaline components have higher negative charge. CONCLUSION: Aconitine and glycyrrhizic acid, aconitine and rhein, berberine and rhein can form macromolecular complexes, ephedrine and glycyrrhizic acid can not.

[Identification and genetic analysis of SNPs in duck adiponectin gene].

Single nucleotide polymorphisms (SNPs) within the duck adiponectin gene were detected by single strand conformation polymorphism (SSCP) using 5 pairs of primers to amplify an area spanning the open reading frame. Eight duck breeds, including Kunshan Sheldrake, Cherry Valley Meat duck, Gaoyou duck, Shanma duck, Jinding duck, Longbai duck, Jingjiang Sheldrake and White feather Muscovy duck, were used. Seven nucleotide variations were found, of which G430A, A457G, and T523C resulted in amino acid changes of A144T, I153V, and Y175H, respectively. The remaining 4 SNPs were C507T, T540C, C576T and C597T. Eight genotypes (AA, AB, AC, BB, BC, CC, DD, and DE) were detected in the 8 breeds. Chi(2) analysis showed that the distribution of the eight genotypes was very different among the different breeds (P < 0.01). Ex-cept for the Jingding duck, all breeds were in accordance with the Hardy-Weinberg equilibrium. Genetic analysis indicated that homozygosity was highest in the Jinding duck, lowest in the Gaoyou duck and similar in other breeds. Polymorphism information content (PIC) was low in the Jinding, high in the Gaoyou and intermediate in other breeds. These results showed that the adiponectin gene had a high level of polymorphism in different duck breeds, and could be used as a candidate gene to analyze the correlation between its polymorphism and fat traits in duck.