MiR-2284b regulation of α-s1 casein synthesis in mammary epithelial cells of dairy goats.

The lactation character of dairy goats is the most important characteristic, and milk protein is an important index to evaluate milk quality. Casein accounts for more than 80% of the total milk protein in goat milk and is the main component of milk protein. Using GMECs (goat mammary epithelial cells) as the research object, the CHECK2 vector of the CSN1S1 gene and the overexpression vector of pcDNA 3.1 were constructed, and the mimics of miR-2284b and the interfering RNA of CSN1S1 were synthesized. Using PCR, RT-qPCR, a dual luciferase activity detection system, EdU, CCK8, cell apoptosis detection and ELISA detection, we explored the regulatory mechanism and molecular mechanism of miR-2284b regulation of αs1-casein synthesis in GMECs. miR-2284b negatively regulates proliferation and apoptosis of GMECs and αs1-casein synthesis. Two new gene sequences of CSN1S1 were discovered. CSN1S1-1/-2 promoted the proliferation of GMECs and inhibited cell apoptosis. However, it had no effect on αs1-casein synthesis. MiR-2284b negatively regulates αs1-casein synthesis in GMECs by inhibiting the CSN1S1 gene. These results all indicated that miR-2284b could regulate αs1-casein synthesis, thus playing a theoretical guiding role in the future breeding process of dairy goats and accelerating the development of dairy goat breeding.

Endoplasmic reticulum stress exacerbates microplastics-induced toxicity in animal cells.

Human and animal exposure to microplastics (MPs) contained in food is inevitable because of their widespread existence in the environment. Nevertheless, MPs toxicity studies in ruminants often lack attention. Here, we assessed the cytotoxicity of polystyrene microplastics (PS MPs) on goat mammary epithelial cells (GMECs). Compared to controls, PS MPs treatment significantly reduced cell viability, altered cell morphology and disrupted organelle integrity. Detection of membrane potential and reactive oxygen species (ROS) suggested that PS MPs induced mitochondrial dysfunction and oxidative stress. Further transcriptome analysis also confirmed alterations in these pathways. In addition, several genes related to endoplasmic reticulum (ER) homeostasis were significantly regulated in the transcriptional profile. Subsequent experiments confirmed that PS MPs induce ER stress via the PERK/eIF2α/CHOP pathway, accompanied by intracellular Ca2+ overload. Meanwhile, downstream activation of the Bax/Bcl-2 pathway and caspase cascade released apoptotic signals, which led to apoptosis in GMECs. Interestingly, the addition of PERK inhibitor (ISRIB) attenuated PS MPs-induced ER stress and apoptosis, which suggests that ER stress may exacerbate PS MPs-induced cytotoxicity. This work reveals the impact of MPs on mammalian cytotoxicity, enriches the mechanisms for the toxicity of MPs, and provides insight for further assessment of the risk of MPs in food.

Combined Transcriptomic and Proteomic of Corynebacterium pseudotuberculosis Infection in the Spleen of Dairy Goats.

Corynebacterium pseudotuberculosis (C. pseudotuberculosis) is a zoonotic chronic infectious disease. It mainly occurs in dairy goats reared in herds, and once it invades the dairy goats, it is difficult to completely remove it, causing great harm to the development of the sheep industry. This study mainly was based on TMT-based quantitative proteomics and RNA-seq methods to measure the spleen samples of infected dairy goats at different time periods. Nine four-month-old dairy goats were divided into three groups, with three goats in each group. The dairy goats in the first group (NC group) were inoculated with 1.0 mL of sterilized normal saline subcutaneously, and the second (72 h group) and third groups (144 h group) were inoculated with 1.0 mL of 1 × 107 cfu/mL bacterial solution subcutaneously in the neck. Significant changes in the protein and mRNA expression were observed in different infection and control groups. In the 72 h group, 85 genes with differential genes and proteins were up-regulated and 91 genes were down-regulated in this study. In the 144 h group, 38 genes with differential genes and proteins were up-regulated and 51 genes were down-regulated. It was found that 21 differentially expressed genes and proteins were co-up-regulated in the two groups. There were 20 differentially expressed genes and proteins which were co-down-regulated in both groups. The 72 h group were mainly enriched in protein processing in the endoplasmic reticulum, lysosome, amino sugar and nucleotide sugar metabolism and the estrogen signaling pathway. In the 144 h group, they were protein processing in the endoplasmic reticulum pathway which was enriched by mRNA-proteins pairs co-upregulated by the five pairs. The combined transcriptomic and proteomic analyses were performed to provide insights into the effects of C. pseudotuberculosis through several regulatory features and pathways. We found that in the early stage of infection (72 h), the co-upregulated gene-protein pairs were enriched in multiple pathways, which jointly defended against a bacterial invasion. However, in the later stages of infection (144 h), when the disease stabilizes, a few co-upregulated gene-protein pairs played a role in protein processing in the endoplasmic reticulum pathway. In addition, the mRNA and protein expressions of dairy goats infected with the bacteria at different periods of time indicated the adaptability of dairy goats to the bacteria. At the same time, it guides us to carry out a corresponding treatment and feeding management for dairy goats according to different periods of time.

Borrowing Strength and Borrowing Index for Bayesian Hierarchical Models.

A novel borrowing strength measure and an overall borrowing index to characterize the strength of borrowing behaviors among subgroups are proposed for a given Bayesian hierarchical model. The constructions of the proposed indexes are based on the Mallow's distance and can be easily computed using MCMC samples for univariate or multivariate posterior distributions. Consequently, the proposed indexes can serve as meaningful and useful exploratory tools to better understand the roles played by the priors in a hierarchical model, including their influences on the posteriors that are used to make statistical inferences. These relationships are otherwise ambiguous. The proposed methods can be applied to both the continuous and binary outcome variables. Furthermore, the proposed approach can be easily adapted to various settings of clinical trials, where Bayesian hierarchical models are deem appropriate. The effectiveness of the proposed method is illustrated using extensive simulation studies and a real data example.

Histogram analysis in predicting the grade and histological subtype of meningiomas based on diffusion kurtosis imaging.

BACKGROUND: Presurgical grading is particularly important for selecting the best therapeutic strategy for meningioma patients. PURPOSE: To investigate the value of histogram analysis of diffusion kurtosis imaging (DKI) maps in the differentiation of grades and histological subtypes of meningiomas. MATERIAL AND METHODS: A total of 172 patients with histopathologically proven meningiomas underwent preoperative magnetic resonance imaging (MRI) and were classified into low-grade and high-grade groups. Mean kurtosis (MK), fractional anisotropy (FA), and mean diffusivity (MD) histograms were generated based on solid components of the whole tumor. The following parameters of each histogram were obtained: 10th, 25th, 75th, and 90th percentiles, mean, median, maximum, minimum, and kurtosis, skewness, and variance. Comparisons of different grades and subtypes were made by Mann-Whitney U test, Kruskal-Wallis test, ROC curves analysis, and multiple logistic regression. Pearson correlation was used to evaluate correlations between histogram parameters and the Ki-67 labeling index. RESULTS: Significantly higher maximum, skewness, and variance of MD, mean, median, maximum, variance, 10th, 25th, 75th, and 90th percentiles of MK were found in high-grade than low-grade meningiomas (all P < 0.05). DKI histogram parameters differentiated 7/10 pairs of subtype pairs. The 90th percentile of MK yielded the highest AUC of 0.870 and was the only independent indicator for grading meningiomas. Various DKI histogram parameters were correlated with the Ki-67 labeling index (P < 0.05). CONCLUSION: The histogram analysis of DKI is useful for differentiating meningioma grades and subtypes. The 90th percentile of MK may serve as an optimal parameter for predicting the grade of meningiomas.

Collateral grade of the Willis' circle predicts outcomes of acute intracranial internal carotid artery occlusion before thrombectomy.

BACKGROUND AND PURPOSE: Endovascular mechanical thrombectomy (EVMT) shows significant promise in improving acute ischemic stroke (AIS) with proximal artery occlusion, but outcomes have been variable. We explored the patients treated by thrombectomy to investigate the association between a favorable clinical outcome of EVMT in intracranial internal carotid artery occlusion (iICAO) and a set of predictors. METHODS: A total of 38 iICAO patients treated by EVMTs were analyzed. Primary collateral grades (PCG) at baseline based on the integrity of Willis' circle were categorized into three degrees. The favorable outcomes, measured by modified Rankin scale (mRS), were defined as ≤2 at 90 days. The reperfusion was one of the most important confounders, defined as modified thrombolysis in cerebral infarction (mTICI) ≥ 2b. The other risk factors included demographic characteristics, vascular risk factors, stroke severity, procedural EVMT, and PCG at baseline was adjusted to reveal the association with favorable outcomes. RESULTS: Of 38 iICAO patients, 65.8% (25 in 38) achieved reperfusion. However, only 31.6% (12/38) achieved favorable outcomes at 90 days. With a PCG3, 61.5% of them achieved favorable outcomes, while only 37.5% of those with PCG2 and PCG1 achieved favorable outcomes (p = .003). In multivariable logistic regression, PCG was revealed as a predictor for favorable outcomes (OR 5.278, p = .019) after adjusting the reperfusion and other factors. CONCLUSIONS: The PCG based on the integrity of Willis' circle might be an underlying predictor of the prognosis of AIS in patients with iICAO after EVMT. The function of intact anterior communicating artery (AcoA) and ipsilateral posterior communicating artery (PcoA) in favoring prognosis of the iICAO patients might need to be validation in future study.

Disease-specific and glucocorticoid-responsive serum biomarkers for Duchenne Muscular Dystrophy.

Extensive biomarker discoveries for DMD have occurred in the past 7 years, and a vast array of these biomarkers were confirmed in independent cohorts and across different laboratories. In these previous studies, glucocorticoids and age were two major confounding variables. In this new study, using SomaScan technology and focusing on a subset of young DMD patients who were not yet treated with glucocorticoids, we identified 108 elevated and 70 decreased proteins in DMD relative to age matched healthy controls (p value < 0.05 after adjusting for multiple testing). The majority of the elevated proteins were muscle centric followed by cell adhesion, extracellular matrix proteins and a few pro-inflammatory proteins. The majority of decreased proteins were of cell adhesion, however, some had to do with cell differentiation and growth factors. Subsequent treatment of this group of DMD patients with glucocorticoids affected two major groups of pharmacodynamic biomarkers. The first group consisted of 80 serum proteins that were not associated with DMD and either decreased or increased following treatment with glucocorticoids, and therefore were reflective of a broader effect of glucocorticoids. The second group consisted of 17 serum proteins that were associated with DMD and these tended to normalize under treatment, thus reflecting physiologic effects of glucocorticoid treatment in DMD. In summary, we have identified a variety of circulating protein biomarkers that reflect the complex nature of DMD pathogenesis and response to glucocorticoids.

Differentiation between vestibular schwannomas and meningiomas with atypical appearance using diffusion kurtosis imaging and three-dimensional arterial spin labeling imaging.

PURPOSE: An accurate differentiation between vestibular schwannomas (VS) and meningiomas is critical in determining treatment strategies and clinical prognoses. However, misdiagnoses may occur when typical imaging appearances are absent. The purpose of this study was to assess the performances of diffusion kurtosis imaging (DKI) and three-dimensional arterial spin labeling imaging (3D-ASL) in the differentiation of VS and meningiomas with atypical appearance. MATERIALS AND METHODS: Thirty-eight patients with pathologically proven VS and meningiomas were consecutively enrolled. All patients had no typical appearance and underwent DKI and 3D-ASL scan. Then, the DKI and 3D-ASL parameters were measured. Statistical analyses were performed using independent-sample t-tests, Mann-Whitney U tests and receiver operating characteristic (ROC) curve analyses. RESULTS: Mean kurtosis (MK), radial kurtosis (RK), axial kurtosis (AK), fractional anisotropy (FA) and cerebral blood flow (CBF) were significantly lower in VS than those in meningiomas, mean diffusivity (MD) were significantly higher in VS than that in meningiomas (all P < 0.05). ROC curve analyses showed that the diagnostic performance of kurtosis values outperformed those of other MR parameters. A cut-off RK value of 0.766 yielded a sensitivity of 91.67%, a specificity of 100.0%, and an accuracy of 94.74%, with an AUC of 0.988. CONCLUSION: DKI and 3D-ASL are useful for differentiating VS and meningiomas with atypical appearance, with kurtosis values of DKI have the best diagnostic efficiency.