Optimizing the Protein Fluorescence Reporting System for Somatic Embryogenesis Regeneration Screening and Visual Labeling of Functional Genes in Cotton

Protein fluorescence reporting systems are of crucial importance to in-depth life science research, providing systematic labeling tools for visualization of microscopic biological activities in vivo and revolutionizing basic research. Cotton somatic cell regeneration efficiency is low, causing difficulty in cotton transformation. It is conducive to screening transgenic somatic embryo using the fluorescence reporting system. However, available fluorescence labeling systems in cotton are currently limited. To optimize the fluorescence reporting system of cotton with an expanded range of available fluorescent proteins, we selected 11 fluorescent proteins covering red, green, yellow, and cyan fluorescence colors and expressed them in cotton. Besides mRuby2 and G3GFP, the other nine fluorescent proteins (mCherry, tdTomato, sfGFP, Clover, EYFP, YPet, mVenus, mCerulean, and ECFP) were stably and intensely expressed in transgenic callus and embryo, and inherited in different cotton organs derive from the screened embryo. In addition, transgenic cotton expressing tdTomato appears pink under white light, not only for callus and embryo tissues but also various organs of mature plants, providing a visual marker in the cotton genetic transformation process, accelerating the evaluation of transgenic events. Further, we constructed transgenic cotton expressing mCherry-labeled organelle markers in vivo that cover seven specific subcellular compartments: plasma membrane, endoplasmic reticulum, tonoplast, mitochondrion, plastid, Golgi apparatus, and peroxisome. We also provide a simple and highly efficient strategy to quickly determine the subcellular localization of uncharacterized proteins in cotton cells using organelle markers. Lastly, we built the first cotton stomatal fluorescence reporting system using stomata-specific expression promoters (ProKST1, ProGbSLSP, and ProGC1) to drive Clover expression. The optimized fluorescence labeling system for transgenic somatic embryo screening and functional gene labeling in this study offers the potential to accelerating somatic cell regeneration efficiency and the in vivo monitoring of diverse cellular processes in cotton.

Can intermittent fasting help in stem-cell rejuvenation?

Background: Human tissues are sustained by stem cells, the balance between stem cell self-renewal and differentiation, and cell death is the crucial element of haemostasis, which plays a vital role in tissue remodelling. Stem cell therapy is recognized as regenerative medicine. We can stimulate stem cell growth/regeneration through various events. Objectives: In this review, we illustrate whether fasting can stimulate stem cell regeneration. Methodology: A literature survey was undertaken to gather recent research and address the effects of fasting on stem cell renewal, which was the review's purpose. Results: Enterocytes in drosophila were restored after exposure to the dietary restriction. As the result, fasting before etoposide exposure safeguarded mice against harm caused by etoposide when compared with the fed group. A study conducted on the effects of periodic fasting in yeast, mice, and humans shows the increase in lifespan and stress resistance of yeast fasting reduces the risks due to age factors and diseases like diabetes, cardiovascular diseases and promotes a healthy life span of mice and human. Conclusion: Dietary limitations will be used in conjunction with the existing therapeutic approach as adjuvant therapy. It will be a more effective treatment. Fasting has the potential to protect against the negative effects of chemotherapy while also boosting stem cell regeneration, according to preclinical findings. Even though regenerative medicine and stem cell therapy are still in their embryonic stage, greater interventions are needed to show the target pathway of fasting in stem cell renewal. Keywords: Fasting, stem cell regeneration, emerging therapy.

Lateral plate mesoderm cell-based organoid system for NK cell regeneration from human pluripotent stem cells

ABSTRACTHuman pluripotent stem cell (hPSC)-induced NK (iNK) cells are a promising “off-the-shelf” cell product for universal immune therapy. Conventional methods for iNK cell regeneration from hPSCs include embryonic body-formation and feeder-based expansion steps, which bring instability, time-consuming, and high costs for manufacture. In this study, we develop an embryonic body-free, organoid aggregate method for NK cell regeneration from hPSCs. In a short time window of 27-day induction, millions of hPSC input can produce over billions of iNK cells without the necessity of NK cell-expansion feeders. The iNK cells highly express classical toxic granule proteins, apoptosis-inducing ligands, as well as abundant activating and inhibitory receptors. Functionally, the iNK cells eradicate human tumor cells by mechanisms of direct cytotoxity, apoptosis, and antibody-dependent cellular cytotoxicity. This study provides a reliable scale-up method for regenerating human NK cells from hPSCs, which promotes the universal availability of NK cell products for immune therapy.

Is Cell Regeneration and Infiltration a Double Edged Sword for Porcine Aortic Valve Deterioration: A Large Cohort of Histopathological Analysis

Background Bioprostheses are the commonest prostheses used for valve replacement in the western world. The major flaw of bioprostheses is the occurrence of structural valve deterioration (SVD). The objective of this study was to assess in a large cohort of patients the pathologic features of porcine aortic valve (PAV) SVD based on histomorphological and immunopathological features.Methods and materials 109 cases of resected PAV were observed grossly and histopathologically. The type and amount of infiltrated cells were evaluated in the different type of bioprosthetic SVD by immunohistochemical staining . Results The most common cause of SVD was calcification, leaflet dehiscence and tear (23.9%,19.3% and 18.3%, respectively). Immunohistochemical staining demonstrated that vimentin positive cells aggregated around the calcified area in calcified PAV. Macrophages infiltrated in the calcified, lacerated and dehiscence PAV. However, MMP-1 expression was mainly found in the lacerated PAV. The VIM(+)/SMA(-) and VIM(+)/CD31(-) cells were found in PAV. The endothelia rate of dehiscence leaflets were higher than that of calcified and lacerated leaflets. A large amount of CD31 positive cells aggregated in the spongy layer in the lacerated and dehiscence PAV. Conclusions Cell regeneration and infiltration is a double edged sword for the PAV deterioration. Valve interstitial cells (VIC) have essential role in PAV calcification. Macrophages infiltration maybe involve in the different type of SVD, but only MMP-1expression involves in leaflets laceration. VIM(+)/CD31(-) valve endothelial cells (VECs) protect the PAV against the formation of calcified and lacerated lesions. The existence of untransformed VECs maybe one of pathologic substrate of PAV tear and dehiscence, although they can prevent VICs activation and subsequent valve fibrosis and calcification.

Combination of GLP-1 Receptor Activation and Glucagon Blockage Promotes Pancreatic β-Cell Regeneration In Situ in Type 1 Diabetic Mice

Pancreatic β-cell neogenesis in vivo holds great promise for cell replacement therapy in diabetic patients, and discovering the relevant clinical therapeutic strategies would push it forward to clinical application. Liraglutide, a widely used antidiabetic glucagon-like peptide-1 (GLP-1) analog, has displayed diverse β-cell-protective effects in type 2 diabetic animals. Glucagon receptor (GCGR) monoclonal antibody (mAb), a preclinical agent that blocks glucagon pathway, can promote recovery of functional β-cell mass in type 1 diabetic mice. Here, we conducted a 4-week treatment of the two drugs alone or in combination in type 1 diabetic mice. Although liraglutide neither lowered the blood glucose level nor increased the plasma insulin level, the immunostaining showed that liraglutide expanded β-cell mass through self-replication, differentiation from precursor cells, and transdifferentiation from pancreatic α cells to β cells. The pancreatic β-cell mass increased more significantly after GCGR mAb treatment, while the combination group did not further increase the pancreatic β-cell area. However, compared with the GCGR mAb group, the combined treatment reduced the plasma glucagon level and increased the proportion of β cells/α cells. Our study evaluated the effect of liraglutide, GCGR mAb monotherapy, or combined strategy in glucose control and islet β-cell regeneration and provided useful clues for the future clinical application in type 1 diabetes.