Loss of NLRP3 reduces oxidative stress and polarizes intratumor macrophages to attenuate immune attack on endometrial cancer

IntroductionThe interaction between endometrial cancer (EMC) cells and intratumoral macrophages plays a significant role in the development of the disease. PYD domains-containing protein 3 (NLRP3) inflammasome formation triggers caspase-1/IL-1β signaling pathways and produces reactive oxygen species (ROS) in macrophages. However, the role of NLRP3-regulated ROS production in macrophage polarization and the subsequent growth and metastasis of EMC remains unknown.MethodsWe conducted bioinformatic analysis to compare NLRP3 levels in intratumoral macrophages from EMC and normal endometrium. In vitro experiments involved knocking out NLRP3 in macrophages to shift the polarization from an anti-inflammatory M1-like phenotype to a proinflammatory M2-like phenotype and reduce ROS production. The impact of NLRP3 depletion on the growth, invasion, and metastasis of co-cultured EMC cells was assessed. We also evaluated the effect of NLRP3 depletion in macrophages on the growth and metastasis of implanted EMC cells in mice.ResultsOur bioinformatic analysis showed significantly lower NLRP3 levels in intratumoral macrophages from EMC than those from normal endometrium. Knocking out NLRP3 in macrophages shifted their polarization to a proinflammatory M2-like phenotype and significantly reduced ROS production. NLRP3 depletion in M2-polarized macrophages increased the growth, invasion, and metastasis of co-cultured EMC cells. NLRP3 depletion in M1-polarized macrophages reduced phagocytic potential, which resulted in weakened immune defense against EMC. Additionally, NLRP3 depletion in macrophages significantly increased the growth and metastasis of implanted EMC cells in mice, likely due to compromised phagocytosis by macrophages and a reduction in cytotoxic CD8+ T cells.DiscussionOur results suggest that NLRP3 plays a significant role in regulating macrophage polarization, oxidative stress, and immune response against EMC. NLRP3 depletion alters the polarization of intratumoral macrophages, leading to weakened immune defense against EMC cells. The reduction in ROS production by the loss of NLRP3 may have implications for the development of novel treatment strategies for EMC

Comparison of Nerve-Sparing Radical Hysterectomy and Radical Hysterectomy: a Systematic Review and Meta-Analysis

Background/Aims: Radical hysterectomy (RH) for the treatment of cervical cancer frequently caused pelvic organ dysfunctions. This study aimed to compare the results of pelvic organ function and recurrence rate after Nerve sparing radical hysterectomy (NSRH) and RH treatment through systematic review and meta-analysis. Methods: PubMed, Web of Science and China Knowledge Resource Integrated Database were searched from inception to 25 February 2015. Studies of cervical cancer which reported radical hysterectomy or nerve sparing radical hysterectomy were included. The quality of included studies was evaluated using the guidelines of Cochrane Handbook for Systematic Reviews of Interventions. Statistical analysis was performed using Review Manager 5.3 software (Cochrane Collaboration). Results: A total of 20 studies were finally included. Meta-analysis demonstrated that NSRH was associated with less bladder and anorectal dysfunction than RH. The time to bladder and anorectal function recovery after NSRH was shorter than RH. Patients undergoing NSRH also scored higher than patients undergoing RH at Female Sexual Function Index (FSFI). On the other hand, the local recurrence and overall recurrence rate were similar between NSRH and RH. Conclusion: NSRH may be an effective technique for lowering pelvic organ dysfunction and improving the function recovery without increasing the recurrence rate of cervical cancer

Spatiotemporal distribution of schistosomiasis transmission risk in Jiangling County, Hubei Province, P.R. China.

ObjectiveThis study aims to explore the spatiotemporal distribution of schistosomiasis in Jiangling County, and provide insights into the precise schistosomiasis control.MethodsThe descriptive epidemiological method and Joinpoint regression model were used to analyze the changes in infection rates of humans, livestock, snails, average density of living snails and occurrence rate of frames with snails in Jiangling County from 2005 to 2021. Spatial epidemiology methods were used to detect the spatiotemporal clustering of schistosomiasis transmission risk in Jiangling county.ResultsThe infection rates in humans, livestock, snails, average density of living snails and occurrence rate of frames with snails in Jiangling County decreased from 2005 to 2021 with statistically significant. The average density of living snails in Jiangling County was spatially clustered in each year, and the Moran's I varied from 0.10 to 0.26. The hot spots were mainly concentrated in some villages of Xionghe Town, Baimasi Town and Shagang Town. The mean center of the distribution of average density of living snails in Jiangling County first moved from northwest to southeast, and then returned from southeast to northwest after 2014. SDE azimuth fluctuated in the range of 111.68°-124.42°. Kernal density analysis showed that the high and medium-high risk areas of Jiangling County from 2005 to 2021 were mainly concentrated in the central and eastern of Jiangling County, and the medium-low and low risk areas were mainly distributed in the periphery of Jiangling County.ConclusionsThe epidemic situation of schistosomiasis decreased significantly in Jiangling County from 2005 to 2021, but the schistosomiasis transmission risk still had spatial clustering in some areas. After transmission interruption, targeted transmission risk intervention strategies can be adopted according to different types of schistosomiasis risk areas

Small extracellular vesicles derived from embryonic stem cells restore ovarian function of premature ovarian failure through PI3K/AKT signaling pathway

Abstract Background Premature ovarian failure (POF) has a great impact on reproductive endocrine function in females, and it is an important cause of infertility. Previous studies have demonstrated that small extracellular vesicles (sEVs) derived from stem cells play an important role in tissue regeneration. This study aimed to investigate the therapeutic effect of sEVs derived from embryonic stem cells (ESCs-sEVs) on damaged ovaries and explore the underlying molecular mechanisms. Methods Mice POF models were established by injecting mice with cyclophosphamide and busulfan. Then, ESCs-sEVs were intravenously transplanted into POF mice. The plasma of mice was harvested at 1 and 2 weeks after treatment to analyze the levels of anti-Mullerian hormone (AMH), estradiol (E2), and follicle stimulating hormone (FSH) by ELISA. The morphology of ovaries and follicles was observed by H&E staining, and apoptosis of granulosa cells was detected by TUNEL. In vitro, EdU and CCK-8 tests were used to evaluate the proliferation of cultured granulosa cells stimulated by ESCs-sEVs. Western blotting was used to determine the expression of PI3K/AKT and apoptotic-related proteins. Results After transplantation of ESCs-sEVs, the levels of serum sex hormones recovered to normal levels. In addition, the number of follicles was significantly increased, and the number of apoptotic cells was decreased. The results in vitro revealed that ESCs-sEVs could significantly improve the proliferation rate of granulosa cells and increase the expression of phosphorylated PI3K and AKT. Meanwhile, the positive effect on proliferation and the negative effect on apoptosis observed in granulosa cells were obviously decreased when the PI3K/AKT signaling pathway was inhibited. Conclusion Our findings suggested that ESCs-sEVs could improve ovarian function by regulating the PI3K/AKT signaling pathway, which could provide a promising clinical therapy for POF

Exosomes secreted by urine-derived stem cells improve stress urinary incontinence by promoting repair of pubococcygeus muscle injury in rats

Abstract Background Previous studies revealed that urine-derived stem cells (USCs) could promote myogenesis after the impairment of the sphincter muscles. However, the effects of exosomes secreted by USCs (USCs-Exo) were not elucidated. Exosomes are nanosized membrane vesicles secreted by the cells. They have been proved to be effective in protecting against tissue injury and therapeutic in tissue repair. USCs are ideal sources of exosomes because of the noninvasive obtaining method and self-renewal abilitiy. This study aimed to show the therapeutic effects of USCs-Exo on improving stress urinary incontinence (SUI). Methods Rat SUI models were established in this study using vaginal balloon inflation, and urodynamic and histological examination were carried out after exosome application. The proliferation and differentiation of muscle satellite cells (SCs) were evaluated using EdU, Cell Counting Kit 8, immunofluorescence staining, and Western blot analysis. mRNAs and proteins related to the activation of SCs were detected by reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) and Western blot analysis. Results After exosome injection, the urodynamic parameters significantly improved and the injured muscle tissue recovered well. The activation, proliferation, and differentiation of SCs were promoted. The phosphorylation of extracellular-regulated protein kinases (ERK) was enhanced. When ERK was inhibited, the promoting effect of USCs-Exo treatment disappeared. Conclusion The findings of this study elucidated the functional roles of USCs-Exo in satellite cell ERK phosphorylation and identified a novel agent for skeletal muscle regeneration, providing a basis for further exploring a cell-free correction for SUI

The Temperature Dependent Proteomic Analysis of <em>Thermotoga maritima</em>

<div><p><em>Thermotoga maritima</em> (<em>T. maritima</em>) is a typical thermophile, and its proteome response to environmental temperature changes has yet to be explored. This study aims to uncover the temperature-dependent proteins of <em>T. maritima</em> using comparative proteomic approach. <em>T. maritima</em> was cultured under four temperatures, 60°C, 70°C, 80°C and 90°C, and the bacterial proteins were extracted and electrophoresed in two-dimensional mode. After analysis of gel images, a total of 224 spots, either cytoplasm or membrane, were defined as temperature-dependent. Of these spots, 75 unique bacterial proteins were identified using MALDI TOF/TOF MS. As is well known, the chaperone proteins such as heat shock protein 60 and elongation factor Tu, were up-regulated in abundance due to increased temperature. However, several temperature-dependent proteins of <em>T. maritima</em> responded very differently when compared to responses of the thermophile <em>T. tengcongensis</em>. Intriguingly, a number of proteins involved in central carbohydrate metabolism were significantly up-regulated at higher temperature. Their corresponding mRNA levels were elevated accordingly. The increase in abundance of several key enzymes indicates that a number of central carbohydrate metabolism pathways of <em>T. maritima</em> are activated at higher temperatures.</p> </div

The temperature-dependent expression of the <i>T. maritima</i> carbohydrate metabolism related genes validated by Western blot and real-time PCR.

<p>A) The temperature-dependent proteins of central carbohydrate pathways in <i>T. maritima</i> were validated by Western blot. The six temperature-dependent proteins in <i>T. maritima</i> were specifically recognized by their correspondent antibodies at the four temperatures. B) The temperature-dependent expression of the T. maritima carbohydrate metabolism related genes were validated by real-time PCR. The experimental procedure and the statistic estimation for the parallel runs were described in “<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0046463#s2" target="_blank">Material and Methods</a>”.</p

The possible network of central carbon metabolism in <i>T. maritima</i>.

<p>The symbols, “↑, ↓, and ↓↑,” indicate the protein abundances up-, down-, or bell-shaped-regulation response to temperature change. <b>Abbreviations: ED</b>, Entner–Doudoroff pathway; <b>EMP</b>, Embden–Meyerhof–Parnas pathway; <b>XynC</b>, family 10 xylanase XynC; <b>Glad</b>, gluconate dehydratase; <b>Kdpg</b>, 2-dehydro-3-deoxyphosphogluconate aldolase; <b>Glk</b>, glucokinase; <b>Gpi</b>, glucose-6-phosphate isomerase; <b>Pfk</b>, phosphofructokinase; <b>Fba</b>, fructose-1,6-bisphosphate aldolase; <b>HK</b>, hexokinase; <b>Pmi</b>, phosphomannoisomerase; <b>Pmm</b>, phosphomannomutase; <b>Tim</b>, triosephosphate isomerase; <b>Glda</b>, glycerol dehydrogenase; <b>Gk</b>, glycerol kinase; <b>Gapdh</b>, D-glyceraldehyde-3-phosphate dehydrogenase; <b>Pgk</b>, 3-phosphoglycerate kinase; <b>Pgm</b>, phosphoglycerate mutase; <b>Eno</b>, enolase; <b>Ppdk</b>, pyruvate,orthophosphate dikinase; <b>Pfor</b>, pyruvate synthase subunit porA; <b>OaDc</b>, oxaloacetate decarboxylase; <b>CoA</b>, coenzyme A; <b>PEP</b>, phosphoenolpyruvate.</p