Gap junction-mediated cell-cell interaction between transplanted mesenchymal stem cells and vascular endothelium in stroke

We have shown previously that transplanted bone marrow mononuclear cells (BM‐MNC), which are a cell fraction rich in hematopoietic stem cells, can activate cerebral endothelial cells via gap junction‐mediated cell‐cell interaction. In the present study, we investigated such cell‐cell interaction between mesenchymal stem cells (MSC) and cerebral endothelial cells. In contrast to BM‐MNC, for MSC we observed suppression of vascular endothelial growth factor uptake into endothelial cells and transfer of glucose from endothelial cells to MSC in vitro. The transfer of such a small molecule from MSC to vascular endothelium was subsequently confirmed in vivo and was followed by suppressed activation of macrophage/microglia in stroke mice. The suppressive effect was absent by blockade of gap junction at MSC. Furthermore, gap junction‐mediated cell‐cell interaction was observed between circulating white blood cells and MSC. Our findings indicate that gap junction‐mediated cell‐cell interaction is one of the major pathways for MSC‐mediated suppression of inflammation in the brain following stroke and provides a novel strategy to maintain the blood‐brain barrier in injured brain. Furthermore, our current results have the potential to provide a novel insight for other ongoing clinical trials that make use of MSC transplantation aiming to suppress excess inflammation, as well as other diseases such as COVID‐19 (coronavirus disease 2019)

The expression of hepatoma upregulated protein in human endometrium during the menstrual cycle.

Aims:Human endometrium resists embryo implantation except during the window period. Currently, uterine HURP expression is known to be involved in endometrial stromal proliferation during embryo implantation of mice. Thus, we demonstrated hepatoma up-regulated protein (HURP) expression in the human endometrium during the menstrual cycle, as well as HURP regulation in endometrial stromal cells (ESCs).Materials and methods:We collected human endometrial samples from different menstrual cycle phases (early/late proliferative, and early/mid/late secretory), and then analyzed these samples by immunohistochemistry, reverse transcription-polymerase chain reaction, and Western blotting. We also assessed the effects of two sex-steroid hormones, 17β-estradiol (E2) and 4-pregnene-3,20-dione (P4) on the cultured stromal cells.Results:HURP protein was localized to the nucleus of the endometrial both epithelial and stromal cells in all stages. Also, HURP mRNA and protein in human endometrial tissue was significantly up-regulated during late-proliferative and secretory phase, compared with early-proliferative phase. In ESCs, HURP expression was regulated by E2, but not P4.Conclusions:We indicated that cyclic changes in HURP expression in human normal ESC strongly suggested up-regulation by estrogen. Taken together, since estrogen responses are fundamental in endometrial biology, uterine expression of HURP may be involved in female reproductive function during the menstrual cycle

Induction of the epithelial-mesenchymal transition in the endometrium by chronic endometritis in infertile patients.

Background:The purpose of the present study was to evaluate the relationship between chronic endometritis and the epithelial-mesenchymal transition in the endometrium of infertile patients in the implantation phase.Methods:Endometrial biopsy specimens from 66 infertility patients were analyzed. The presence of chronic endometritis was investigated by immunostaining for CD138. Immunohistochemical staining for E-cadherin, N-cadherin, Slug, and Snail was performed, and the expression profiles were statistically analyzed according to the presence of chronic endometritis. When the loss of E-cadherin expression and/or the positive expression of N-cadherin was detected, the specimen was considered epithelial-mesenchymal transition-positive. Epithelial-mesenchymal transition-positive cases were also statistically analyzed according to the presence of chronic endometritis. The characteristics of the patients in the epithelial-mesenchymal transition-positive and epithelial-mesenchymal transition-negative groups were compared. The association between variables, including age, body mass index, gravidity, parity, and each causative factor of infertility and epithelial-mesenchymal transition positivity was analyzed.Results:The rates of the loss of E-cadherin expression, the gain of N-cadherin and epithelial-mesenchymal transition positivity were significantly higher in chronic endometritis patients. The expression of Slug, cytoplasmic Snail, and nuclear Snail was also detected at significantly higher rates in chronic endometritis patients. Chronic endometritis were related to the epithelial-mesenchymal transition.Conclusion:The epithelial-mesenchymal transition was frequently detected in the endometrium in infertile patients with chronic endometritis. Since the epithelial-mesenchymal transition is associated with chronic endometritis, the epithelial-mesenchymal transition appears to be involved in the alteration of mechanisms of implantation

卵巣組織凍結保存を実施した1歳11ヵ月の女児の一例

　がん治療の発達により小児がんを克服する患者が増加している一方で、治療による晩期合併症として妊孕能の低下が惹起されることがある。今回、1歳の女児に対して妊孕能温存療法として、卵巣組織凍結保存を行ったので報告する。　症例は1歳11ヵ月。腹部膨満を主訴に近医を受診したところ、骨盤内腫瘤を認めた。諸検査により仙骨部原発の卵黄嚢腫瘍と診断された。BEP療法後、残存腫瘍に大量化学療法を施行する可能性があり、妊孕能温存の相談のため当科へ紹介された。親権者の同意を得て腹腔鏡下右付属器切除術、卵巣組織凍結保存を施行した。摘出卵巣の大部分が原始卵胞を有する皮質であり、組織を卵巣長軸に対して垂直に細切し、卵巣皮質と髄質を合わせて凍結保存した。術後1日目より発熱を認め、抜管時の嘔吐による誤嚥性肺炎と診断し治療を行った。　1歳児の妊孕能温存療法であったため治療に対する同意、および手術操作への配慮が必要であった。卵巣組織の構造が思春期以降のものと異なるため、組織の凍結方法を工夫した。幼児との意思疎通は困難で、慎重に術後のバイタルおよび身体所見を観察すること、家族や小児科スタッフとの情報共有、連携が重要であると考えられた。(著者抄録

Relationship of Chronic Endometritis With Chronic Deciduitis in Cases of Miscarriage.

Background: The presence of chronic deciduitis (CD) was determined in patients diagnosed with or without chronic endometritis (CE) before pregnancy.Objective: To study the effect of CE on decidua in cases of miscarriage.Methods:Decidual tissue was obtained from the patients who miscarried at the first pregnancy within a year after the diagnosis of the presence or absence of CE. The number and distribution pattern of plasma cells stained with CD138 in decidual tissue in 10 high-power fields (HPFs) was examined. The prevalence of CD diagnosed with four different grade; grade 0, no plasma cell in 10 HPFs, thus Non-CD;grade 1, rare single plasma cells; grade 2, rare clusters or more than 5 single cells total; and grade 3, many plasma cells with more than 5 clusters, were examined and compared between Non-CE and CE.Results:The incidence rate of CD of grade2 + 3 was significantly higher in CE than Non-CE (53.8%; 7/13 vs. 0%; 0/13, P < 0.01). Presence of clusters or a number of plasma cells in 10 HPFs of decidua showed a sensitivity of 53.8%, a specificity of 100%, a positive predictive value of 100%, and a negative predictive value of 68.4% for the diagnosis of CE.Conclusion:Presence of clusters of plasma cells or five or more of plasma cells in decidua was found in more than half of CE, but not found in Non-CE. When CD with cluster or five or more of plasma cells is confirmed histologically in miscarriage decidual tissue, the presence of CE before the pregnancy should be suspected

Histological diagnostic criterion for chronic endometritis based on the clinical outcome.

Background:The diagnostic criteria of chronic endometritis remain controversial in the treatment for infertile patients.Methods: A prospective observational study was conducted in a single university from June 2014 to September 2017. Patients who underwent single frozen-thawed blastocyst transfer with a hormone replacement cycle after histological examination for the presence of chronic endometritis were enrolled. Four criteria were used to define chronic endometritis according to the number of plasma cells in the same group of patients: 1 or more (≥ 1) plasma cells, 2 or more (≥ 2), 3 or more (≥ 3), or 5 or more (≥ 5) in 10 high-power fields. Pregnancy rates, live birth rates, and miscarriage rates of the non-chronic endometritis and the chronic endometritis groups defined with each criterion were calculated. A logistic regression analysis was performed for live births using eight explanatory variables (seven infertility factors and chronic endometritis). A receiver operating characteristic curve was drawn and the optimal cut-off value was calculated.Results:A total of 69 patients were registered and 53 patients were finally analyzed after exclusion. When the diagnostic criterion was designated as the presence of ≥ 1 plasma cell in the endometrial stroma per 10 high-power fields, the pregnancy rate, live birth rate, and miscarriage rate were 63.0% vs. 30.8%, 51.9% vs. 7.7%, and 17.7% vs. 75% in the non-chronic and chronic endometritis groups, respectively. This criterion resulted in the highest pregnancy and live birth rates among the non-chronic endometritis and the smallest P values for the pregnancy rates, live birth rates, and miscarriage rates between the non-chronic and chronic endometritis groups. In the logistic regression analysis, chronic endometritis was an explanatory variable negatively affecting the objective variable of live birth only when chronic endometritis was diagnosed with ≥ 1 or ≥ 2 plasma cells per 10 high-power fields. The optimal cut-off value was obtained when one or more plasma cells were found in 10 high-power fields (sensitivity 87.5%, specificity 64.9%).Conclusions:Chronic endometritis should be diagnosed as the presence of ≥ 1 plasma cells in 10 high-power fields. According to this diagnostic criterion, chronic endometritis adversely affected the pregnancy rate and the live birth rate

NEAT1 long noncoding RNA regulates transcription via protein sequestration within subnuclear bodies

Paraspeckles are subnuclear structures formed around nuclear paraspeckle assembly transcript 1 (NEAT1)/MEN epsilon/beta long noncoding RNA (IncRNA). Here we show that paraspeckles become dramatically enlarged after proteasome inhibition. This enlargement is mainly caused by NEAT1 transcriptional up-regulation rather than accumulation of undegraded paraspeckle proteins. Of interest, however, using immuno-electron microscopy, we find that key paraspeckle proteins become effectively depleted from the nucleoplasm by 50% when paraspeckle assembly is enhanced, suggesting a sequestration mechanism. We also perform microarrays from NEAT1-knockdown cells and find that NEAT1 represses transcription of several genes, including the RNA-specific adenosine deaminase B2 (ADARB2) gene. In contrast, the NEAT1-binding paraspeckle protein splicing factor proline/glutamine-rich (SFPQ) is required for ADARB2 transcription. This leads us to hypothesize that ADARB2 expression is controlled by NEAT1-dependent sequestration of SFPQ. Accordingly, we find that ADARB2 expression is strongly reduced upon enhanced SFPQ sequestration by proteasome inhibition, with concomitant reduction in SFPQ binding to the ADARB2 promoter. Finally, NEAT1(-/-) fibroblasts are more sensitive to proteasome inhibition, which triggers cell death, suggesting that paraspeckles/NEAT1 attenuates the cell death pathway. These data further confirm that paraspeckles are stress-responsive nuclear bodies and provide a model in which induced NEAT1 controls target gene transcription by protein sequestration into paraspeckles