Genome-scale Transcriptomic Insights Into the Gene-regulatory Network of Seed Abortion in Triploid Siraitia Grosvenorii

BackgroundSiraitia grosvenorii (Swingle) C. Jeffrey, also known as Luohanguo or monk fruit, is a famous traditional Chinese medicine ingredient with important medicinal value and broad development prospects. Diploid S. grosvenorii has too many seeds. Thus, studying the molecular mechanism of seed abortion in triploid S. grosvenorii, identifying the abortion-related genes, and regulating their expression will be a new direction to obtain seedless S. grosvenorii. Herein, we examined the submicroscopic structure of triploid S. grosvenorii seeds during abortion. ResultsBy measuring the content of endogenous hormones, we found that abscisic acid (ABA) and trans-zeatin (ZR) levels were significantly downregulated at days 15 and 20 after flowering. RNA-seq of triploid seeds at different developmental stages was performed to identify the key genes regulating abortion of triploid S. grosvenorii seeds. Multiple differentially expressed genes between adjacent stages were identified; seven genes were differentially expressed across all stages. Weight gene co-expression network analysis revealed that the enhancement of monoterpene and terpene metabolic processes might lead to seed abortion by reducing substrate flow to ABA and ZR.ConclusionsThese findings provide insights into the gene-regulatory network of seed abortion in triploid S. grosvenorii from different perspectives, thereby facilitating the innovation of the breeding technology of S. grosvenorii.

Genome-Wide Identification of TCP Transcription Factors Family in Sweet Potato Reveals Significant Roles of miR319-Targeted TCPs in Leaf Anatomical Morphology

Plant-specific TCP transcription factors play vital roles in the controlling of growth, development, and the stress response processes. Extensive researches have been carried out in numerous species, however, there hasn’t been any information available about TCP genes in sweet potato (Ipomoea batatas L.). In this study, a genome-wide analysis of TCP genes was carried out to explore the evolution and function in sweet potato. Altogether, 18 IbTCPs were identified and cloned. The expression profiles of the IbTCPs differed dramatically in different organs or different stages of leaf development. Furthermore, four CIN-clade IbTCP genes contained miR319-binding sites. Blocking IbmiR319 significantly increased the expression level of IbTCP11/17 and resulted in a decreased photosynthetic rate due to the change in leaf submicroscopic structure, indicating the significance of IbmiR319-targeted IbTCPs in leaf anatomical morphology. A systematic analyzation on the characterization of the IbTCPs together with the primary functions in leaf anatomical morphology were conducted to afford a basis for further study of the IbmiR319/IbTCP module in association with leaf anatomical morphology in sweet potato.

Microscopic and ultramicroscopic structure of typical chordae tendineae of the human’s left ventricle bicuspid valve

Objective – to determine the features of the micro- and submicroscopic structure of typical chordae tendineae of the mitral valve of the human heart.Materials and methods. This study was made on preparations of 35 human hearts, in particular the chordae tendineae of the bicuspid valves of the left ventricles of children and people of early and middle adulthood using the methods of light and electron microscopy.Results. The study of typical chordae tendineae of the mitral valve leaflets was made using the microscopic method and the method of electron microscopy. The areas where the chordae tendineae diverge from the papillary muscles, the middle third of the chordae tendineae, and the place of attachment of the chordae tendineae to the mitral valve leaflets were studied. Chordae tendineae are covered with endothelium from all sides. A peripheral collagen-elastic layer of loose fibrous connective tissue is localized under the endothelium, along the entire perimeter of the chordae tendineae and it differs in structure in different parts of the chordae at the submicroscopic level. Bundles of densely packed collagen fibers, which are oriented rectilinearly along the chordae, form the core of the chordae tendineae. Bundles of striated cardiac typical muscle cells are localized in areas where the chordae tendineae origin from the papillary muscles. It was found that in the composition of the chordae tendineae of children the cellular component in the fibrous connective tissue predominates during comparing the typical chordae tendineae of different aged people.Conclusions. This morphological study made it possible to obtain new information and to clarify the data that are already known to morphologists on the micro- and submicroscopic structure of typical chordae tendineae of the mitral valve of the human heart. We found that differences in the histological structure are more typical for the areas of origin of typical chordae tendineae from the papillary muscles and areas of their attachment to the leaflets of the mitral valve. The diameter of the core of the chordae tendineae increases and the thickness of the loose peripheral collagen-elastic layer decreases with age. The received data can serve as a theoretical basis for the further comparative morphological and clinical studies of the mitral valve of the human heart. That, in turn, will facilitate to an increasing the level of diagnostic, to improve methods of prevention and treatment of minor heart anomalies, congenital and acquired defects of the valvular apparatus of the heart.

Restricting Glutamine Uptake Enhances NSCLC Sensitivity to Third-Generation EGFR-TKI Almonertinib

The emergence of secondary resistance is the main failure cause of epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) as a targeted therapy for non-small cell lung cancer (NSCLC). EGFR mutations of NSCLC cells can markedly increase glutamine transporter (SLC1A5) expression, thereby increasing glutamine metabolism. Glutamine metabolites can activate EGFR downstream signals, including mTOR, ERK1/2, STAT3, etc., which is an important cause for the decreased sensitivity of NSCLC to EGFR-TKIs. CCK8 and Annexin V/PI assays were conducted to detect the effects of Almonertinib and/or V9302 on the proliferation and apoptosis of NSCLC cells. Proteomics was used to determine the effect of Almonertinib on energy metabolism-related proteins in NSCLC. siRNA transfection was performed to study the effect of SLC1A5 down-regulation on cell proliferation. In addition, the effects of drugs on colony formation capacity were determined by colony formation assay. Immunofluorescence and Western blot were utilized to detect the apoptosis- and autophagy-related proteins expression. DAPI staining was utilized to detect the effect of drugs on the nucleus. Transmission electron microscope was used to observe the changes of submicroscopic structure such as autophagosomes and nucleus of cells. mCherry-GFP-LC3B tandem fluorescent protein was to used to detect the level of autophagy flux. Tumor-bearing nude mouse model was utilized to detect the effect of V9302 on the anti-tumor effect of Almonertinib in vivo. As a result, Almonertinib suppressed H1975 and A549 cell proliferation depended on its dosage and treatment duration, and it also induced apoptosis. A549 cells with wild-type EGFR had lower sensitivity to Almonertinib. The expression of SLC1A5 was up-regulated by stimulating with low concentration of Almonertinib in NSCLC cells. SLC1A5 was highly expressed in A549 cells with wild-type EGFR. Glutamine deletion or SLC1A5 inhibition/silencing inhibited the proliferation of NSCLC cells, and decreased cellular glutamine uptake. The combination of SLC1A5 inhibitor V9302 and Almonertinib had a synergistic inhibitory effect on the proliferation of NSCLC. V9302 enhanced the effect of Almonertinib in apoptosis-inducing in NSCLC cells. The combination of V9302 and Almonertinib might induce apoptosis by inhibiting autophagy.

A Deep Insight of Spermatogenesis and Hormone Levels of Aqua-Cultured Turbot (Scophthalmus maximus)

Turbot (Scophthalmus maximus) is an important marine fish both in Europe and North China. Although there are plenty of studies on the reproduction of turbot, the complete cytological process of spermatogenesis remains unclear. In this study, we investigated the submicroscopic structure of total 23 types of male germ cells throughout the breeding season, with a relatively complete process of the primary spermatocytes. We found that the spermatid tail formed early at Spermatid II, and there were at least 16 spherical mitochondria in the spermatozoa. The hepatosomatic index (HSI) and gonadosomatic index (GSI) both peaked during the breeding season. Preliminary analysis showed that the vitality of mature sperm was negatively correlated with the proportion of sperm deformity. The serum 3,5,3′-triiodothyronine (T3), 5-hydroxytryptamine (5-HT), testosterone (T), 17α,20β-Dihydroxy-4-pregnen-3-one (17α,20β-DHP), and 17β-estradiol (E2) all increased during the maturity period, with the change of T content most noticeable. Whereas in the testis, an overall high level of 11-ketotestosterone (11-KT) was more remarkable. The expression and localization of androgen receptor (AR) mRNA showed that the AR was highly expressed at the stages of II (15 – 70 g), with a slight rebound at the mature stages [IV(2200 g) to V(2500 g)], whose change was ahead to the changes of T and 11-KT. Fluorescence in situ hybridization (FISH) analysis showed that the AR mainly distributed in but not limited to Sertoli cells. This study represents the most complete overview of the reproductive cycle and spermatogenesis of turbot, which provides an important reference for the reproduction research and the guidance of flatfish breeding.

Response of Lamiophlomis rotata (Benth.) Kudo to degradation of alpine grasslands in the western Sichuan plateau, China

Background: Lamiophlomis rotata (Benth.)Kudo, also known as Duyiwei in China, is considered to be one of the most common Tibetan medicine. It is famous for the significant effects on improving blood circulation and relieving pain. Due to the environmental deterioration and excessive consumption, the wild resources of L.rotata is decreasing year by year. However, its artificial cultivation has not been realized. This study aimed to initially explore the adaptation mechanism of L.rotata to degraded alpine grassland, and provide reference for its wild tending and artificial cultivation.Methods: In this study, we used a sample method to conduct a population ecological survey of plants and collect plant leaf samples from mildly, moderately, and extremely degraded grassland. The content of SOD, POD, CAT, PRO, MDA, SS, SP, carotene, chlorophyll A and chlorophyll B in different sample leaves was measured by colorimetry. Microscopic characteristics of the cross-section, epidermis and submicroscopic structure of the leaves were observed and compared.Results: The density of L. rotata in moderately degraded grassland was the highest，where L. rotata had the strongest resistance. And the analysis of seven physiological indicators combined with stress resistance showed that the average content of MDA, SOD, POD, CAT and SS is the highest in L. rotata of the moderately degraded grassland. The microscopic observation showed that, by increasing the thickness of the epidermis, the density of the glandular scales, the proportion of palisade tissue and the number of gland scales, L. rotata was able to adapt to the degraded grassland. Meanwhile, morphological studies have shown that L. rotata could enhance root growth in the degraded grassland. Furthermore, in response to the degradation of alpine grassland on the Qinghai-Tibet Plateau, L. rotata population and biomass distribution in the underground parts have increased. However, L. rotata does not have a competitive advantage in mildly degraded grassland. Compare with the mildly and moderately habitat, the population of L. rotata is small in the extremely degraded habitat.Conclusion: The results showed that among the 3 levels of degraded grassland, L. rotata in the moderately degraded grassland shows the strongest resistance. This study can provide a reference for breeding and ecological planting of L. rotata, suggests that in the nursery stage, the medium degraded grassland should be chosen for ecological breeding to enhance the survival rate of seedlings. While after large-scale planting, reasonable community interventions, like the P. fruticosa+F. rubra community, should be built to simulate the yield of medicinal materials.

Comparison analysis of Bacillus Calmette-Guerin induced Autophagy mechanisms in Macrophages Derived from Human Induced Pluripotent Stem Cells and THP-1

Background Tuberculosis (TB) remains a major global public health problem and the leading cause of mortality by a single infectious agent. TB is a chronic infectious disease that is primarily caused by Mycobacterium tuberculosis (Mtb). Macrophage (Mφ) are the main hosts of Mtb, the interaction between Mtb and Mφ plays an important role in the pathogenesis of TB.Summary The macrophages used in the current study are mostly derived from tumor cell lines or peripheral blood mononuclear cells (PBMC), but the application of such cells still have many problems needed to be sloved, such as the loss of function due to changes in genetic structure and the difficulty in cell acquisition. Human induced pluripotent stem cells (hiPS) represent an innovative source for the standardized in vitro generation of Mφ, and show novel promise in exploring disease pathogenesis, particularly TB. Current studies have revealed that autophagy plays a central role in the interaction between Mtb and Mφ, but the molecular mechanism involoved remains unclear and the exact role of hiPS-derived macrophages (hiPS-Mφ) in regulating autophagy induced by Mtb also remains unclear. To investigate the similarities and differences in hiPS-Mφ and THP-1-Mφ in anti-tuberculosis immunity, this study successfully obtained macrophages derived from hiPS and THP-1, then explored the mechanism behind Bacillus Calmette-Guerin (BCG)-induced autophagy through transcriptome sequencing analysis, qPCR, Western Blot Analysis and cell submicroscopic structure observation etc.. Our findings revealed that BCG infection of hiPS-Mφ and THP-1-Mφ would promote autophagy by regulating the expression of autophagy-related genes, which also indicated that the BCG-induced autophagy in hiPS-Mφ and THP-1-Mφ may be associated with PI3K/AKT/mTOR signaling pathway. However, there are some differences in the mechanism by which BCG infects macrophages from different sources and induces autophagy. Considering the above findings, we have provided novel insights into the role of macrophages along with autophagy in the anti-tuberculosis immune mechanism and the possibility of establishing an in vitro hiPS-Mφ-TB disease model.

Microscopic and submicroscopic structure of the heart atria and auricles in condition of the experimental thermal trauma

Burns is the damage of the skin or mucous membranes (often with underlying tissues), because of high-temperature influence (thermal burns), chemical, electrical, radiation, combined, thermochemical and electrothermal factors influence. According to the investigations, burns rank third among other injuries, and in some countries, second, second only to transport injuries. Although the level of survival after burns is increasing, the problem of their treatment remains relevant in modern medicine and pharmacy. The aim was to study the peculiarities and the patterns of morphofunctional changes in the structural components of the heart atria and auricles in case of the experimental thermal trauma. The study involves 30 adult guinea pigs with a bodyweight of 630-670 g. The burn was applied under general ether anesthesia with water vapor at a temperature of 96-97 0C on the surface of the skin of the animal's back for 60 seconds. The size of the lesion area was 18-20% of the body surface. The depth of the lesion corresponded to the degree ІІІА -IIIB. The experimental animals were divided into two groups: the first - intact guinea pigs (6 heads); second - animals with thermal trauma (24 heads). Were used a number of methods of investigation, including macrometric and massometric - to establish the structural restructuring of the heart and its parts, microscopic and electron microscopic to detect changes in the structural components of the atrium and auricle of the heart; morphometric - to obtain quantitative parameters of the morphological components of the heart; mathematical and statistical - to ensure the analysis of the reliability of the research results. In intact guinea pigs, the morphological organization of the heart auricles differs from the atria by the direction of the muscle fibers, the ratio of morphometric parameters of their structural components and the content of muscle endocrine cells. Submicroscopically, endocrine myocytes of the auricles of the heart contain more hormonal granules than atria. In the ears of the heart, the relative volume of connective tissue is 1,34 times greater, and the relative volume of muscle fibers is 1,03 times smaller than in the atria. Submicroscopically, endocrine muscle cells of the heart auricle contain more hormonal granules than atria. It has been established that thermal trauma causes significant morphofunctional changes in the atria and auricles of the heart at different levels of their structural organization. The degree of damage depends on the duration of the experiment and develops against the background of increased plasma toxicity. In the stages of burn shock and early toxemia, adaptive-compensatory processes develop in the structures of the heart, and irreversible destructive changes occur in the stages of late toxemia and septicotoxemia.

Microstructure of oxide ferromanganese ores in the World ocean as the proof of their bacterial origin

The study of submicroscopic structure of oxide ores revealed their similarity to the present-day bacterial communities. It is shown that the structure of cobalt-bearing crusts and ferromanganese nodules is based on bacterial mats, which permits identifying them as stromatolites and oncolites. The facts in favor of intense interaction between biofilms and the environment are found. The signs of mineral phase formation are registered as a result of biochemical absorption and assimilation of iron and manganese by bacteria.

Deoxynivalenol Induces Inflammatory Injury in IPEC-J2 Cells via NF-κB Signaling Pathway

The aim of this study was to investigate the effects of deoxynivalenol (DON) exposure on the inflammatory injury nuclear factor kappa-B (NF-κB) pathway in intestinal epithelial cells (IPEC-J2 cells) of pig. The different concentrations of DON (0, 125, 250, 500, 1000, 2000 ng/mL) were added to the culture solution for treatment. The NF-κB pathway inhibitor pyrrolidine dithiocarbamate (PDTC) was used as a reference. The results showed that when the DON concentration increased, the cell density decreased and seemed damaged. With the increase of DON concentration in the culture medium, the action of diamine oxidase (DAO) in the culture supernatant also increased. The activities of IL-6, TNF-α, and NO in the cells were increased with the increasing DON concentration. The relative mRNA expression of IL-1β and IL-6 were increased in the cells. The mRNA relative expression of NF-κB p65, IKKα, and IKKβ were upregulated with the increasing of DON concentration, while the relative expression of IκB-α mRNA was downregulated. At the same time, the expression of NF-κB p65 protein increased gradually in the cytoplasm and nucleus with a higher concentration of DON. These results showed that DON could change the morphology of IPEC-J2 cells, destroy its submicroscopic structure, and enhance the permeability of cell membrane, as well as upregulate the transcription of some inflammatory factors and change the expression of NF-κB-related gene or protein in cells.