ABSTRACT
With the rapid development of gene editing technology, the study of spermatogonial stem cells (SSCs) holds great significance in understanding spermatogenesis and its regulatory mechanism, developing transgenic animals, gene therapy, infertility treatment and protecting rare species. Biogenesis of lysosome-related organelles complex 1 subunit 1 (BLOC1S1) is believed to have anti-brucella potential. Exploring the impack of BLOC1S1 on goat SSCs not only helps investigate the ability of BLOC1S1 to promote SSCs proliferation, but also provides a cytological basis for disease-resistant breeding research. In this study, a BLOC1S1 overexpression vector was constructed by homologous recombination. The BLOC1S1 overexpression cell line of goat spermatogonial stem cells was successfully constructed by lentivirus packaging, transfection and puromycin screening. The overexpression efficiency of BLOC1S1 was found to be 18 times higher using real time quantitative PCR (RT-qPCR). Furthermore, the results from cell growth curve analysis, flow cytometry for cell cycle detection, and 5-ethynyl-2'-deoxyuridine (EdU) staining showed that BLOC1S1 significantly increased the proliferation activity of goat SSCs. The results of RT-qPCR, immunofluorescence staining and Western blotting analyses revealed up-regulation of proliferation-related genes (PCNA, CDK2, CCND1), and EIF2S3Y, a key gene regulating the proliferation of spermatogonial stem cells. These findings strongly suggest that the proliferative ability of goat SSCs can be enhanced through the EIF2S3Y/ERK pathway. In summary, this study successfully created a goat spermatogonial stem cell BLOC1S1 overexpression cell line, which exhibited improved proliferation ability. This research laid the groundwork for exploring the regulatory role of BLOC1S1 in goat spermatogonia and provided a cell platform for further study into the biological function of BLOC1S1. These findings also establish a foundation for breeding BLOC1S1 overexpressing goats.
Subject(s)
Animals , Male , Goats , Stem Cells , Spermatogonia/metabolism , Cell Proliferation , Flow Cytometry , Testis/metabolismABSTRACT
Sepsis is a systemic disease with severe health consequences, and it was redefined in 2016 as a life-threatening organ dysfunction caused by an abnormal host response to infection and is a global public health priority. In recent years, there has been increasing recognition of the role of dysregulated micronutrient iron metabolism in the pathogenesis of sepsis. The concept of ferroptosis, an iron-dependent, non-apoptotic mode of cell death characterized by the accumulation of lipid reactive oxygen species (ROS), was first proposed by Dixon et al. in 2012. As a novel mode of programmed cell death, ferroptosis differs in morphological and biochemical characteristics from various forms of cell death, such as apoptosis, autophagy, necrosis and lysis. Recent studies have shown that ferroptosis plays an important regulatory role in the development of sepsis and has become a research focus and highlight for the diagnosis and prognosis of related diseases. Therefore, this paper reviews the latest developments in ferroptosis in sepsis, in order to further understanding its pathogenesis and providing new therapeutic targets for sepsis-related organ dysfunction.
ABSTRACT
Objective To use the real-time shear wave elastography(SWE)to detect the elasticmodulus values of different breast lesions,and to assess its application value in differential diagnosis of benign and malignant breast lesions.Methods SWE was per-formed in 71 breast lesions from 63 pathologically confirmed patients.The maximum,minimum and mean elasticmodulus values in breast lesions were detected and the ROC curves were used to determine the optimal diagnostic cut off value.Results Among 7 1 le-sions,48 cases were benign lesions and 23 cases were malignant lesions.The diagnostic accuracy and Youden index of the ordinary ultrasound were 77.5% and 0.55 respectively.The maximum,minimum and mean elasticity values for malignant lesions were (106.7±37.9),(23.7±6.4),(44.8±6.6)kPa respectively,whereas which for benign lesions were(42.7±14.6),(17.2±6.3), (29.4±8.0)kPa respectively.The maximum,minimum and mean elastic values had statistically significant differences between be-nign and malignant lesions(P<0.05).The areas under the curve(AUC)of the maximum,minimum and mean elastic values were 0.963,0.798 and 0.914 respectively.Taking 78.1 kPa as the threshold of the maximum elasticity value,the sensitivity was 82.6%and the specificity was 95.8%.Taking 37.6 kPa as the threshold of the mean elasticity value,the sensitivity was 87.0% and the specificity was 83.3%.Conclusion SWE provides a new way for the differential diagnosis of benign and malignant breast lesions, and the maximum and mean elastic value can be used as the diagnostic evidence for differentiating benign and malignant breast le-sions.