Stromal Niche Signals That Orchestrate Intestinal Regeneration.

Stromal cell populations have a central role in providing signals that support the maintenance, differentiation, and function of the intestinal epithelium. The behavior and fate of epithelial cells is directed by the spatial organization of stromal cells that either sustain stem and progenitor cell identity or drive differentiation. A combination of single-cell analyses, mouse models, and organoid coculture assays have provided insight into the diversity of signals delivered by stromal cells. Signaling gradients are established and fine-tuned by the expression of signaling agonists and antagonists along the crypt-villus axis. On epithelial injury, there are disruptions to the abundance and organization of stromal populations. There are also distinct changes in the signals originating from these cells that impact remodeling of the epithelium. How these signals coordinate to mediate epithelial repair or sustain tissue injury in inflammatory bowel diseases is beginning to emerge. Understanding of these processes may lead to opportunities to target stromal cell populations as a strategy to modify disease states.

Germline and Somatic Cell Syncytia in Insects.

Syncytia are common in the animal and plant kingdoms both under normal and pathological conditions. They form through cell fusion or division of a founder cell without cytokinesis. A particular type of syncytia occurs in invertebrate and vertebrate gametogenesis when the founder cell divides several times with partial cytokinesis producing a cyst (nest) of germ line cells connected by cytoplasmic bridges. The ultimate destiny of the cyst's cells differs between animal groups. Either all cells of the cyst become the gametes or some cells endoreplicate or polyploidize to become the nurse cells (trophocytes). Although many types of syncytia are permanent, the germ cell syncytium is temporary, and eventually, it separates into individual gametes. In this chapter, we give an overview of syncytium types and focus on the germline and somatic cell syncytia in various groups of insects. We also describe the multinuclear giant cells, which form through repetitive nuclear divisions and cytoplasm hypertrophy, but without cell fusion, and the accessory nuclei, which bud off the oocyte nucleus, migrate to its cortex and become included in the early embryonic syncytium.

Arsenic exposure impairs intestinal stromal cells.

Arsenic is a toxicant commonly found in drinking water. Even though its main route of exposure is oral, little is known of the impact of in vivo arsenic exposure on small intestine. In vitro studies have shown that arsenic decreases differentiation of stem and progenitor cells in several different tissues. Thus, small intestinal organoids were used to assess if arsenic exposure would also impair intestinal stem cell differentiation. Unexpectedly, no changes in markers of differentiated epithelial cells were seen. However, exposing mice to 100 ppb arsenic in drinking water for 5 weeks impaired distinct populations of intestinal stromal cells. Arsenic reduced the width of the pericryptal lamina propria by 1.6-fold, and reduced Pdgfra mRNA expression, which is expressed in intestinal telocytes and trophocytes, by 4.2-fold. The height or extension of Pdgfra+ telopodes into the villus tip was also significantly reduced. Transcript expression of several other stromal cell markers, such as Grem1, Gli, CD81, were reduced by 1.9-, 2.3-, and 1.4-fold, respectively. Further, significant correlations exist between levels of Pdgfra and Gli1, Grem1, and Bmp4. Our results suggest arsenic impairs intestinal trophocytes and telocytes, leading to alterations in the Bmp signaling pathway.

Rhynchophylline improves trophocyte mobility potential by upregulating ZEB1 level via the inhibition of miR-141-3p level.

Preeclampsia (PE) is characterized by the impaired invasive ability of trophocytes, which can be modulated by microRNAs (miRs). In the current study, the effects of rhynchophylline (Rhy) on the viability and invasive ability of trophocytes were explored by focusing on miR-141-3p/ZEB1 axis. The level of miR-141-3p was modulated in human trophocytes and the changes in cell viability, apoptosis, invasive ability, and ZEB1 level were detected. Then the trophocytes with miR-141-3p overexpression were treated with Rhy and the effects on trophocyte phenotypes were assessed. The induced miR-141-3p level suppressed cell viability, induced apoptosis, and inhibited invasion and ZEB1 level in trophocytes. The treatment of Rhy restored the viability and invasive ability of trophocytes under the overexpression of miR-141-3p, indicating the protective effects of Rhy on trophocytes. The findings in the current study highlighted the protective effects of Rhy on trophocytes during PE progression, which was associated with the inhibition of miR-141-3p.

The cellular niche for intestinal stem cells: a team effort.

The rapidly self-renewing epithelium in the mammalian intestine is maintained by multipotent intestinal stem cells (ISCs) located at the bottom of the intestinal crypt that are interspersed with Paneth cells in the small intestine and Paneth-like cells in the colon. The ISC compartment is also closely associated with a sub-epithelial compartment that contains multiple types of mesenchymal stromal cells. With the advances in single cell and gene editing technologies, rapid progress has been made for the identification and characterization of the cellular components of the niche microenvironment that is essential for self-renewal and differentiation of ISCs. It has become increasingly clear that a heterogeneous population of mesenchymal cells as well as the Paneth cells collectively provide multiple secreted niche signals to promote ISC self-renewal. Here we review and summarize recent advances in the regulation of ISCs with a main focus on the definition of niche cells that sustain ISCs.

Tissue Resident Memory γδT Cells in Murine Uterus Expressed High Levels of IL-17 Promoting the Invasion of Trophocytes.

γδT cells are non-conventional T cells and serve as the bridge for connecting the innate and adaptive immune systems. γδT cells form a substantial population at barrier sites and play an important role in the development of physiology, inflammation, autoimmune diseases and tumors. γδT cells not only distribute in the maternal-fetal interface during pregnancy but also in non-pregnant uterus. However, the phenotypes and functions of γδT cells in uterus were not clear. In the current study, we found that the percentages of γδT cells were significantly higher in uterus than peripheral blood and most of γδT cells in uterus were distributed in endometrium. Further studies indicated that the majority of γδT cells in uterus were memory cells with higher expression of CD44 and CD27 but lower expression of CD62L and CCR7 compared to those in blood. In addition, we found that γδT cells in uterus were tissue resident memory γδT cells expressing CD69, expressed high levels of CCR6, GranzymeB and CD107a. Moreover, γδT cells in uterus were activated and fully expressed transcription factor RORγt. After short time of activation, γδT cells in uterus significantly expressed high levels of IL-17 but not IFN-γ, which promotes the invasion of murine trophocytes. Taken together, our study will lay the foundation for future research on uterine γδT cells in pregnancy and autoimmune disease.

AP2γ mediated downregulation of lncRNA LINC00511 as a ceRNA suppresses trophoblast invasion by regulating miR-29b-3p/Cyr61 axis.

BACKGROUND: Long noncoding RNA LINC00511 has been identified to be aberrant expression and may as a tumor oncogene in various carcinomas. However, the potential role of LINC00511 in the onset of Preeclampsia (PE) pathogenesis remains unexplored. METHODS: Placental tissues from patients with PE were collected to detect expression levels of LINC00511 by qRT-PCR. Human HTR-8/SVneo trophoblast cell line was cultured, CCK-8 assay, wound healing assay and transwell assay were performed to determine the regulation of trophoblast biological function by LINC00511. Bioinformatics analysis, chromatin immunoprecipitation (ChIP), luciferases reporter assay were performed to verify the regulatory mechanism of LINC00511. RESULTS: LINC00511 was aberrantly down-regulated in placental tissues of PE patients. Overexpression of LINC00511 promoted trophoblast cell proliferation, migration and invasion. The transcription factor AP2γ directly binds to the promoter region of LINC00511 to activate transcription. In addition, LINC00511 was enriched in cytoplasm and functioned as a molecular spong for miR-29b-3p, antagonizing its ability to repress Cyr61 protein translation. CONCLUSION: This study demonstrated that AP2γ mediated downregulation of LINC00511 suppresses trophoblast invasion by regulating miR-29b-3p/ Cyr61 axis.

Characterization of fat body cells at different developmental stages of Culex pipiens.

The fat body, originates from mesoderm, has many metabolic functions which changes as the embryonic development of the insect progresses. It plays an important role in the intermediate metabolism and in the metabolism of proteins, lipids and carbohydrates. It has roles in synthesis, absorption and storage of nutrients from hemolymph. It is also responsible for the production of immunological system components, antibacterial compounds and blood clotting proteins. The most common type of fat body cells are trophocytes (the basic cells of the fat body) and oenocytes are found associated with the fat body. In this study, it is aimed at determining the cell types contained in the fat body of Culex pipiens at different developmental stages as well as identifying the molecules such as carbohydrate, protein and lipid contained in each of these cells. Knowing the regional distribution of the fat body cells and the concentration of its content at each developmental stage is important in understanding the process related to its physiology and it may help in fighting against the pest C. pipiens, which is a vector species for many contagious diseases observed in humans and other species. To achieve our goal, we have employed different histochemical techniques (fixatives and staining methods) for staining C. pipiens preparates of different developmental stages and analyzed the structure of the fat body, its distribution, its cell types and the macromolecular contents of the cells. We only observed trophocytes and oenocytes as fat body components in C. pipiens. The trophocytes had all the three macromolecules (lipids, proteins, carbohydrates) in the cytoplasm varying in concentration between the different regions and different stages. The oenocytes were observed below the integument as well as between the muscles in the larvae of Culex pipiens. They were present either as single cells or in clusters and also varied in size. Their cytoplasm was stained strongly for proteins when bromophenol blue staining was applied, but it was rather heterogeneous due to the lipid inclusions. On the contrary, oenocytes were not observed among the adult C. pipiens preparations.

Overexpression of heparanase is associated with preeclampsia by inhibiting invasion of trophocytes.

BACKGROUND: Preeclampsia is associated with inadequate invasion of trophocytes and spiral artery remodeling. As a ß-D-glucuronidase enzyme, Heparanase is related to tumor angiogenesis, development and invasion. Trophocytes have similar characteristics to tumor cells, and heparanase could therefore play an important role in the pathogenesis of preeclampsia. METHODS: The expression of heparanase in severe preeclampsia and normal placentas was detected via real-time PCR, immunohistochemistry and western blotting. The effects of heparanase on trophocytes migration and invasion were investigated by culturing the HTR-8/Svneo cell line with recombinant human heparanase protein in vitro. RESULTS: The levels of inactive 65-kDa heterologous heparanase dimers were obviously increased, and the content of the 50-kDa active polypeptide was decreased in severe preeclampsia. Furthermore, exogenous heparanase protein could reduce the migration and invasion of HTR-8/Svneo cells. CONCLUSION: Our results suggested that heparanase might be an important factor in the pathogenesis of severe preeclampsia.

Ovary of Matsucoccus pini (Insecta, Hemiptera, Coccinea: Matsucoccidae): morphology, ultrastructure, and phylogenetic implications.

The structure of ovary in a representative of the scale insect family Matsucoccidae, Matsucoccus pini, is described at the ultrastructural level. The paired ovaries of M. pini are composed of about 50 ovarioles of telotrophic type that develop asynchronously. An individual ovariole consists of an anterior tropharium (trophic chamber) and posterior vitellarium. The tropharium encloses trophocytes (nurse cells) and early previtellogenic oocytes termed arrested oocytes. In the vitellarium from 1 to 6, linearly arranged oocytes may develop. Analysis of serial sections has shown that each ovariole contains 32 germ cells (trophocytes, arrested oocytes, and developing oocytes). In the cytoplasm of all these cells, small rod-shaped bacteria are present. In the early vitellogenic oocytes, accessory nuclei arise. As vitellogenesis progresses, these nuclei migrate toward the cortical ooplasm. The obtained results are discussed in a phylogenetic context.

Clinical and pathological analysis of 34 cases with exaggerated placental site / 中国医师进修杂志

Objective To summarize the clinical and pathological features of exaggerated placental site (EPS),explore its pathogenesis regularity,diagnosis and treatment strategies.Methods The clinical data related to 34 patients with EPS were analyzed retrospectively.Results In 34 patients,11 patients performed full-term cesarean section,2 patients performed normal vaginal delivery,the other 21 patients had abortion.Thirty-one patients had pregnancy history.Fifteen patients performed hysterectomy,13 patients performed dilatation and curettage,6 patients performed exploratory hysteroscopy and lesions resection.All the patients survived after treatment.Conclusions The patients can not be diagnosed in the antepartum and intrapartum,but can be diagnosed relying on the pathological diagnosis.When it is ineffective to stop bleeding after delivery or abortion by conventional treatment,we should consider the possibility of EPS.Timely perform hysterectomy is theeffective method to stop bleeding,can save the life of patients.If bleeding is not much,curettage or exploratory hysteroscopy can get a significant effective treatment and avoid hysterectomy.

A comparative study of fat body morphology in five mosquito species

The insect fat body plays major roles in the intermediary metabolism, in the storage and transport of haemolymph compounds and in the innate immunity. Here, the overall structure of the fat body of five species of mosquitoes (Aedes albopictus, Aedes fluviatilis, Culex quinquefasciatus, Anopheles aquasalis and Anopheles darlingi) was compared through light, scanning and transmission electron microscopy. Generally for mosquitoes, the fat body consists of lobes projecting into the haemocoel and is formed by great cell masses consisting of trophocytes and oenocytes. Trophocytes are rich in lipid droplets and protein granules. Interestingly, brown pigment granules, likely ommochromes, were found exclusively in the trophocytes located within the thorax and near the dorsal integument of Anopheles, which is suggestive of the role these cells play in detoxification via ommochrome storage. This study provides a detailed comparative analysis of the fat body in five different mosquito species and represents a significant contribution towards the understanding of the structural-functional relationships associated with this organ.

Changes in the fat body during the post-embryonic development of the predator Toxorhynchites theobaldi (Dyar & Knab) (Diptera: Culicidae)

Several studies have focused on understanding the biochemistry and morphology of the fat body of the hematophagous mosquito Aedes aegypti (L.) (Diptera: Culicidae). In contrast, few studies, if any, have focused on morphological characters of the fat body in other mosquitoes, especially non-hematophagous taxa such as the culicid Toxorhynchites. Larvae of Toxorhynchites prey upon the larvae of other mosquito species and are used in vector mosquito control. We investigated aspects of the fat body trophocytes, including the morphometric analyses of the lipid droplets, protein granules and nuclei, during Toxorhynchites theobaldi (Dyar & Knab) post-embryonic development. Following the body weight increase from larval stage L2 to L4, the size of lipid droplets within the trophocytes also increase, and are likely the result of lipogenesis. Lipid droplets decrease in size during L4 to the female pupal stage and increase once again during the period from newly-emerged to mature adult females. Protein granules are observed for the first time in female pupae, and their appearance might be related to protein storage during metamorphosis. The size of the nucleus of trophocytes also increases during larval development, followed by a decrease during metamorphosis and an additional increase as adult female ages. In conclusion, the morphology of the fat body of T. theobaldi changes according to the developmental stage. Our study provides for the first time important insights into T. theobaldi fat body development and contributes to understand this species biology.

Antagonism of Bushen Yiqi Prescription on apoptosis of human first trimester trophoblastic cell by regulating Bcl-2 and XIAP in vitro / 中华中医药杂志

Objective:To investigate the effect and mechanism of Bushen Yiqi Prescription on apoptosis of human first trimester trophoblasts.Methods:Isolated from first trimester placental tissues,human trophoblasts cultured in media with 5%, 10%,20%drug-containing serum for a 24,48 or 72h period.Then we measured cell viability by MTT assay,cell DNA content by flow cytomester,cell Caspase-3 enzyme activity by Caspase-3 active assay kit,and,cell Fas,FasL,Bcl-2 and XIAP protein.Results:Absorbency of trophoblast in 490nm wavelength increased greatly after being incubated with drug-containing serum for 24,48,and 72 h(P