FANCD2 deficiency sensitizes SHH medulloblastoma to radiotherapy via ferroptosis.

Radiotherapy is one of the standard therapeutic regimens for medulloblastoma (MB). Tumor cells utilize DNA damage repair (DDR) mechanisms to survive and develop resistance during radiotherapy. It has been found that targeting DDR sensitizes tumor cells to radiotherapy in several types of cancer, but whether and how DDR pathways are involved in the MB radiotherapy response remain to be determined. Single-cell RNA sequencing was carried out on 38 MB tissues, followed by expression enrichment assays. Fanconi anemia group D2 gene (FANCD2) expression was evaluated in MB samples and public MB databases. The function of FANCD2 in MB cells was examined using cell counting assays (CCK-8), clone formation, lactate dehydrogenase activity, and in mouse orthotopic models. The FANCD2-related signaling pathway was investigated using assays of peroxidation, a malondialdehyde assay, a reduced glutathione assay, and using FerroOrange to assess intracellular iron ions (Fe2+ ). Here, we report that FANCD2 was highly expressed in the malignant sonic hedgehog (SHH) MB subtype (SHH-MB). FANCD2 played an oncogenic role and predicted worse prognosis in SHH-MB patients. Moreover, FANCD2 knockdown markedly suppressed viability, mobility, and growth of SHH-MB cells and sensitized SHH-MB cells to irradiation. Mechanistically, FANCD2 deficiency led to an accumulation of Fe2+ due to increased divalent metal transporter 1 expression and impaired glutathione peroxidase 4 activity, which further activated ferroptosis and reduced proliferation of SHH-MB cells. Using an orthotopic mouse model, we observed that radiotherapy combined with silencing FANCD2 significantly inhibited the growth of SHH-MB cell-derived tumors in vivo. Our study revealed FANCD2 as a potential therapeutic target in SHH-MB and silencing FANCD2 could sensitize SHH-MB cells to radiotherapy via inducing ferroptosis. © 2024 The Pathological Society of Great Britain and Ireland.

Surfactants Enhanced Soil Arsenic Phytoextraction Efficiency by Pteris vittata L.

Soil arsenic (As) pollution has become a global problem. It is urgent to improve the phytoextraction efficiency of soil As. This study found chemical activators (Span 80/SDS and GSH/Span 80/SDS) that can significantly improve the availability of As and the phytoextraction efficiency of As by Pteris vittata L. in As-contaminated soil. Compared with the control, in the soil screening experiment, Span 80/SDS and GSH/Span 80/SDS significantly increased available As in soil by 73.4% and 81.4%, respectively. And in the soil pot experiment, the Span 80/SDS and GSH/Span 80/SDS significantly increased the As concentration in the pinnae of Pteris vittata L. by 53.4% and 41.2%, respectively, and the total As amount extraction by Pteris vittata L. increased significantly by 31.7% and 94.2%, respectively. The results suggest that adding Span 80/SDS and GSH/Span 80/SDS to As-contaminated soil can be considered as an effectively method to improve the efficiency of phytoextraction.

Hymenobacter edaphi sp. nov., isolated from abandoned arsenic-contaminated farmland soil.

A Gram-stain-negative, aerobic, rod-shaped, non-motile, pink-pigmented bacterium, designated NLT, was isolated from arsenic-contaminated farmland soil. Strain NLT showed the highest 16S rRNA gene sequence similarities with those of Hymenobacter jeollabukensis 1-3-3-8T (98.9 %), Hymenobacter gummosus ANT-18T (97.5 %), Hymenobacter paludis KBP-30T (97.4 %), Hymenobacter ocellatus Myx2105T (97.1 %) and Hymenobacter coalescens WW84T (96.4 %). The values of genomic orthoANI and dDDH between strain NLT and Hymenobacter jeollabukensis KCTC 52741T was 90.5 and 41.2 %, respectively, and those between strain NLT and Hymenobacter gummosus KCTC 52166T was 84.4 and 28.4 %, respectively. Strain NLT exhibited DNA-DNA hybridisation values of 41.3 and 44.1 % with Hymenobacter paludis KCTC 32237T and Hymenobacter ocellatus DSM 11117T, respectively. Strain NLT had major fatty acids (>10 %) of summed feature 4 (iso-C17 : 1 I and/or anteiso-C17 : 1 B), iso-C15 : 0 and anteiso-C15 : 0 and the predominant polyamine of homospermidine. The only respiratory quinone was menaquinone-7. The polar lipids were phosphatidylethanolamine, phospholipid, three unidentified lipids and two amino lipids. Strain NLT had a genome size of 6.04 Mb and the average G+C content of 65.6 %. Compared to the other Hymenobacter spp., strain NLT is different in polar lipid profile (without aminophospholipid) and leucine arylamidase activity. Based on the data of the polyphasic analysis, it is considered that strain NLT represented a novel species of genus Hymenobacter, for which the name Hymenobacter edaphisp. nov. is proposed. The type strain is NLT (=KCTC 62521T=CCTCC AB 2018028T).

Responses of water quality and phytoplankton assemblages to remediation projects in two hypereutrophic tributaries of Chaohu Lake.

Water shortages and the presence of point and diffuse source pollution have caused a serious deterioration in water quality in two tributaries (the Tangxi River and Shiwuli River) of Chaohu Lake, China. To reduce nutrient pollution and suppress harmful algal blooms (HABs), hard engineering and ecological remediation projects were implemented. A post-project investigation from 2013 to 2016 was carried out to evaluate the outcome of the remediation projects by monitoring the seasonal and spatial variations in water quality and the phytoplankton community. In the Tangxi River, the average total phosphorus (TP) concentrations in the four seasons were below 0.5 mg L-1, with the lowest concentration (0.29 ±â€¯0.12 mg L-1) found in autumn. Remediation measures including sediment dredging, riparian buffer zone creation, downstream wetland park construction, and water augmentation using reclaimed water and filtered lake water might combine to promote P source mitigation. Moreover, the percentage of bloom-forming cyanobacteria (i.e., Microcystis, Aphanizomenon, Anabaena, Oscillatoria, Phormidium and Planktothrix) in the phytoplankton assemblage and the biomass of the dominant species indicated successful HAB control. In the Shiwuli River, water quality improvements and phytoplankton responses have been observed since 2015 after the upgrading of a local wastewater treatment plant (WWTP) with effluent that was used for flow augmentation. Nevertheless, there is still room for improvement via increasing the river self-purification ability (e.g., the creation of downstream wetlands and riparian buffer zones) and promoting water augmentation according to the experience gained in the remediation projects of the Tangxi River.

Performance evaluation of wastewater treatment using horizontal subsurface flow constructed wetlands optimized by micro-aeration and substrate selection.

The effects of micro-aeration and substrate selection on domestic sewage treatment performance were explored using three pairs (with or without micro-aeration) of horizontal subsurface flow (HSSF) constructed wetlands (CWs) filled with zeolite, ceramsite or quartz granules. The individual and combined effects of micro-aeration and substrate selection on the purification performance of the experimental-scale HSSF CWs were evaluated. The results showed that micro-aeration significantly increased the treatment efficiencies for chemical oxygen demand, total nitrogen, total phosphorus (TP), ortho-phosphate (PO4(3-)-P) and ammonium nitrogen (NH4+-N) using HSSF CWs, while the substrate selection significantly affected the TP, PO4(3-)-P and NH4+-N removal efficiencies (p<0.05). A two-way analysis of variance (ANOVA) indicated that there was a significant interaction term (i.e. micro-aeration×substrate selection) for NH4+-N removal (p<0.05). Among the three substrates, ceramsite was the best substrate for the treatment of domestic sewage using HSSF CWs. Therefore, the results of this study suggest that a ceramsite-filled HSSF CW with micro-aeration could be the optimal configuration for decentralized domestic sewage treatment.

Clinical characteristics and diagnostic confirmation of Internet addiction in secondary school students in Wuhan, China.

AIM: This study investigated the clinical characteristics of internet addiction using a cross-sectional survey and psychiatric interview. METHODS: A structured questionnaire consisted of demographics, Symptom Checklist 90, Self-Rating Anxiety Scale, Self-Rating Depression Scale, and Young's Internet Addiction Test (YIAT) was administered to students of two secondary schools in Wuhan, China. Students with a score of 5 or higher on the YIAT were classified as having Internet Addiction Disorder (IAD). Two psychiatrists interviewed students with IAD to confirm the diagnosis and evaluate their clinical characteristics. RESULTS: Of a total of 1076 respondents (mean age 15.4 ± 1.7 years; 54.1% boys), 12.6% (n = 136) met the YIAT criteria for IAD. Clinical interviews ascertained the Internet addiction of 136 pupils and also identified 20 students (14.7% of IAD group) with comorbid psychiatric disorders. Results from multinomial logistic regression indicated that being male, in grade 7-9, poor relationship between parents and higher self-reported depression scores were significantly associated with the diagnosis of IAD. CONCLUSION: These results advance our understanding of the clinical characteristics of Internet addiction in Chinese secondary school students and may help clinicians, teachers, and other stakeholders better manage this increasingly serious mental condition.

circ_0134120: a new frontier in understanding postmenopausal osteoporosis pathogenesis.

Postmenopausal osteoporosis (OP) is a prevalent skeletal disease with not fully understood molecular mechanisms. This study aims to investigate the role of circular RNA (circRNA) in postmenopausal OP and to elucidate the potential mechanisms of the circRNA-miRNA-mRNA regulatory network. We obtained circRNA and miRNA expression profiles from postmenopausal OP patients from the Gene Expression Omnibus database. By identifying differentially expressed circRNAs and miRNAs, we constructed a circRNA-miRNA-mRNA network and identified key genes associated with OP. Further, through a range of experimental approaches, including dual-luciferase reporter assays, RNA pull-down experiments, and qRT-PCR, we examined the roles of circ_0134120, miR-590-5p, and STAT3 in the progression of OP. Our findings reveal that the interaction between circ_0134120 and miR-590-5p in regulating STAT3 gene expression is a key mechanism in OP, suggesting the circRNA-miRNA-mRNA network is a potential therapeutic target for this condition.

Toxic effects and comparison of common amino antioxidants (AAOs) in the environment on zebrafish: A comprehensive analysis based on cells, embryos, and adult fish.

The ubiquity of amino antioxidants (AAOs) in the environment has attracted increasing attention, given their potential toxicity. This investigation represents a pioneering effort, systematically scrutinizing the toxicological effects of four distinct AAOs across the developmental spectrum of zebrafish, encompassing embryonic, larvae, and adult stages. The results indicate that four types of AAO exhibit varying degrees of cell proliferation toxicity. Although environmentally relevant concentrations of AAOs exhibit a comparatively circumscribed impact on zebrafish embryo development, heightened concentrations (300 µg/L) of N-(1,3-dimethylbutyl)-N'-phenyl-p-phenylenediamine (6PPD) and N-isopropyl-N'-phenyl-p-phenylenediamine (IPPD) distinctly evoke developmental toxicity. Behavioral analysis results indicate that at concentrations of 20 and 300 µg/L, the majority of AAOs significantly reduced the swimming speed and activity of larvae. Moreover, each AAO triggers the generation of reactive oxygen species (ROS) in larvae, instigating diverse levels of oxidative stress. The study delineates parallel toxicological patterns in zebrafish exposed to 300 µg/L of 6PPD and IPPD, thereby establishing a comparable toxicity profile. The comprehensive toxicity effects among the four AAOs is as follows: IPPD >6PPD > N-Phenyl-1-naphthylamine (PANA) > diphenylamine (DPA). These findings not only enrich our comprehension of the potential hazards associated with AAOs but also provide data support for structure-based toxicity prediction models.

Global analysis of the adverse effects of micro- and nanoplastics on intestinal health and microbiota of fish.

The wide occurrence of micro- and nanoplastics (MPs/NPs) within aquatic ecosystems has raised increasing concerns regarding their potential effects on aquatic organisms. However, the effects of MPs/NPs on intestinal health and microbiota of fish remain controversial, and there is a lack of comprehensive understanding regarding how the impact of MPs/NPs is influenced by MPs/NPs characteristics and experimental designs. Here, we conducted a global analysis to synthesize the effects of MPs/NPs on 47 variables associated with fish intestinal health and microbiota from 118 studies. We found that MPs/NPs generally exerted obvious adverse effects on intestinal histological structure, permeability, digestive function, immune and oxidative-antioxidative systems. By contrast, MPs/NPs showed slight effects on intestinal microbial variables. Further, we observed that the responses of intestinal variables to MPs/NPs were significantly regulated by MPs/NPs characteristics and experimental designs. For instance, polyvinyl chloride plastics showed higher toxicity to fish gut than polyethylene and polystyrene did. Additionally, larval fish appeared to be more sensitive to MPs/NPs than juvenile fish. Collectively, this study highlights the potential impacts of MPs/NPs on intestinal health and microbiota of fish, and underscores the determinant role of MPs/NPs characteristics and experimental designs in MPs/NPs toxicity.

Effects of silver nanoparticles on denitrification and anammox in sediments of hypertrophic and mesotrophic lakes.

The abundant production and wide usage of silver nanoparticles (Ag NPs) inevitably lead to their release into aquatic ecosystems. However, it is still unclear about how Ag NPs influence denitrification and anammox (DA) in freshwater sediments. To address this, the sediments of hypertrophic and mesotrophic lakes were exposed to 0.5 and 50 mg/L Ag NPs under anaerobic conditions for 7 days to explore the effects of Ag NPs on environmental variables, including redox potential (Eh), pH, organic matter (OM) and acid volatile sulfide (AVS), and the resulting influence on DA. Experimental results indicated that NO3--N and NH4+-N levels were increased by the low (p > 0.05) and high doses of Ag NPs (p < 0.05) in comparison with the non-Ag control, revealing an inhibitive impact on DA. The level of total nitrogen (TN) was notably increased by the low and high doses of Ag NPs (p < 0.05), perhaps due to inhibited enzyme activity and corresponding encoding gene abundance, which were related to generating gaseous nitrogen such as N2O and N2. In addition, environmental factor Eh was significantly raised by Ag NPs (p < 0.05), further inhibiting DA. Moreover, the quantitative analysis unveiled that denitrifying and anammox bacteria in hypertrophic lakes evinced a stronger resistance to Ag NPs toxicity than those in mesotrophic lakes. Overall, our study revealed that short-term exposure to Ag NPs could inhibit DA in sediments. These findings provide an understanding enabling evaluation and prediction of the environmental risks of Ag NPs in freshwater lakes.

Identification of key modules and candidate genes associated with endometriosis based on transcriptome data via bioinformatics analysis.

BACKGROUNDS: Endometriosis is a common disease in women, but the signaling pathways and genes involved remain unclear. This study screened genes that were differentially expressed in ectopic endometrium (EC) and eutopic endometrium (EU) in endometriosis and provided clues for subsequent experimental verification. METHODS: Endometriosis samples were harvested from inpatients that underwent surgery from 2017 to 2019 with pathological evidence of endometriosis. We assessed the mRNA expression profiles in endometriosis and further conducted gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, Gene Set Enrichment Analysis (GSEA) and weighted gene co-expression network analysis (WGCNA) to identify potential biomarkers in endometriosis. Finally, we further validated hub genes using public databases and immunohistochemistry assays. RESULTS: The upregulated DEGs of ectopic endometrium from endometriosis patients were mainly involved in cell adhesion, MAPK signaling, PI3K-Akt signaling pathways, cytokine receptor interactions, and epithelial-mesenchymal transformation (EMT)-associated signaling pathways. The downregulated DEGs between ectopic endometrium and eutopic endometrium were related to decidualization-associated genes in endometriosis. The correlated gene modules in eutopic endometrial cells were mainly enriched in cell adhesion, embryo implantation and inflammation. The eutopic and ectopic endometrial lesions in endometriosis were involved in the EMT process. Furthermore, we identified 18 co-expression modules during WGCNA analysis. Hub genes in the pale turquoise module were FOSB, JUNB, ATF3, CXCL2, FOS, etc. Significantly enriched KEGG pathways included the TNF, MAPK, foxO, oxytocin, and p53 signaling pathways. Enrichment pathways were directly related to immune surveillance, stem cell self-renewal, and epithelial-mesenchymal transformation. Several pathways and modules of endometriosis are related to cancer-associated pathways, which substantiates the correlation between endometriosis and various gynecological tumors. CONCLUSIONS: Endometriosis was tightly correlated with EMT and fibrosis mediated by inflammatory immunity, cytokines, estrogen, kinases and protooncogene through transcriptomics. Overall, our findings lay the groundwork for understanding the pathogenesis of endometriosis and its relationship with malignant transformation.

Vitamin D3 attenuates SARS-CoV-2 nucleocapsid protein-caused hyperinflammation by inactivating the NLRP3 inflammasome through the VDR-BRCC3 signaling pathway in vitro and in vivo.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection-caused coronavirus disease 2019 (COVID-19) is a global crisis with no satisfactory therapies. Vitamin D3 (VD3) is considered a potential candidate for COVID-19 treatment; however, little information is available regarding the exact effects of VD3 on SARS-CoV-2 infection and the underlying mechanism. Herein, we confirmed that VD3 reduced SARS-CoV-2 nucleocapsid (N) protein-caused hyperinflammation in human bronchial epithelial (HBE) cells. Meanwhile, VD3 inhibited the NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome activation in N protein-overexpressed HBE (HBE-N) cells. Notably, the inhibitors of caspase-1, NLRP3, and NLRP3 or caspase-1 small interference RNA (siRNA) enhanced VD3-induced NLRP3 inflammasome inactivation, with subsequent suppression of interleukin-6 (IL6) and IL1ß release in HBE-N cells, which were abolished by the NLRP3 agonist. Moreover, VD3 increased NLRP3 ubiquitination (Ub-NLRP3) expression and the binding of the VDR with NLRP3, with decreased BRCA1/BRCA2-containing complex subunit 3 (BRCC3) expression and NLRP3-BRCC3 association. VD3-induced Ub-NLRP3 expression, NLRP3 inflammasome inactivation, and hyperinflammation inhibition were improved by the BRCC3 inhibitor or BRCC3 siRNA, which were attenuated by the vitamin D receptor (VDR) antagonist or VDR siRNA in HBE-N cells. Finally, the results of the in vivo study in AAV-Lung-enhanced green fluorescent protein-N-infected lungs were consistent with the findings of the in vitro experiment. In conclusion, VD3 attenuated N protein-caused hyperinflammation by inactivating the NLRP3 inflammasome partially through the VDR-BRCC3 signaling pathway.

Pristine and sulfidized ZnO nanoparticles alter microbial community structure and nitrogen cycling in freshwater lakes.

Zinc oxide nanoparticles (ZnO NPs) and its sulfidized form (ZnS NPs) are increasingly entering into freshwater systems through multiple pathways. However, their impacts on the composition and function of sedimentary microbial communities are still largely unknown. Here, two kinds of lake-derived microcosms were constructed and incubated with ZnO NPs, or ZnS NPs to investigate the short-term (7 days) and long-term (50 days) impacts on sedimentary microbial communities and nitrogen cycling. After 7 days, both ZnO NPs and ZnS NPs dosed microbial communities experienced distinct alterations as compared to the undosed controls. By day 50, the structural shifts of microbial communities caused by ZnO NPs were significantly enlarged, while the microbial shifts induced by ZnS NPs were largely resolved. Additionally, ZnO NPs and ZnS NPs could significantly alter nitrogen species and nitrogen cycling genes in sediments, revealing their non-negligible impacts on nitrogen cycling processes. Furthermore, our data clearly indicated that the impacts of ZnO NPs and ZnS NPs on nitrogen cycling differed distinctly in different lake-derived microcosms, and the impacts were significantly correlated with microbial community structure. Overall, this research suggests that the entrance of pristine or sulfidized ZnO NPs into freshwater systems may significantly impact the sedimentary microbial community structure and nitrogen cycling.

PLXDC2 enhances invadopodium formation to promote invasion and metastasis of gastric cancer cells via interacting with PTP1B.

Plexin-domain containing 2 (PLXDC2) has been reported as an oncoprotein in several human malignancies. However, its expression and roles in gastric cancer remain largely unclear. In this study, we found that PLXDC2 was highly expressed in gastric cancer tissues, and the expression levels were positively correlated with clinicopathological features, but negatively with the patients' outcome. Cox regression analysis identified PLXDC2 as an independent prognostic indicator for the patients. Knockdown of PLXDC2 markedly suppressed the in vitro invasion and in vivo metastasis of gastric cancer cells, while overexpression of PLXDC2 resulted in opposite effects. Mechanistically, PLXDC2 enhanced the level of phosphorylated Cortactin (p-Cortactin) by physically interacting with protein tyrosine phosphatase 1B (PTP1B), an important dephosphorylase, to prevent its dephosphorylating of p-Cortactin, thereby promoting the formation of invadopodia. Collectively, our results indicate that PLXDC2 contributes to the invasion and metastasis of gastric cancer by inhibiting PTP1B to facilitate the invadopodium formation, and may serve as a potential prognostic biomarker and a therapeutic target for this disease.

Elevated Kir2.1/nuclear N2ICD defines a highly malignant subtype of non-WNT/SHH medulloblastomas.

Medulloblastoma (MB) is one of the most common childhood malignant brain tumors (WHO grade IV), traditionally divided into WNT, SHH, Group 3, and Group 4 subgroups based on the transcription profiles, somatic DNA alterations, and clinical outcomes. Unlike WNT and SHH subgroup MBs, Group 3 and Group 4 MBs have similar transcriptomes and lack clearly specific drivers and targeted therapeutic options. The recently revised WHO Classification of CNS Tumors has assigned Group 3 and 4 to a provisional non-WNT/SHH entity. In the present study, we demonstrate that Kir2.1, an inwardly-rectifying potassium channel, is highly expressed in non-WNT/SHH MBs, which promotes tumor cell invasion and metastasis by recruiting Adam10 to enhance S2 cleavage of Notch2 thereby activating the Notch2 signaling pathway. Disruption of the Notch2 pathway markedly inhibited the growth and metastasis of Kir2.1-overexpressing MB cell-derived xenograft tumors in mice. Moreover, Kir2.1high/nuclear N2ICDhigh MBs are associated with the significantly shorter lifespan of the patients. Thus, Kir2.1high/nuclear N2ICDhigh can be used as a biomarker to define a novel subtype of non-WNT/SHH MBs. Our findings are important for the modification of treatment regimens and the development of novel-targeted therapies for non-WNT/SHH MBs.

Factors Influencing Residents' Psychological Status During Standardized Training in COVID-19.

Objectives: To explore the influencing factors of residents' psychological status during standardized training in COVID-19 for finding ways to promote their mental health. Methods: A total of 760 residents were surveyed with a structured questionnaire. Correlation analysis was used to analyze the influencing factors of psychological status of the residents, and a mediation model was constructed to verify the mediating role of satisfaction. Results: Age, willingness to study medicine, and satisfaction were positively correlated with negative psychological status (P < 0.05). And gender, only child or not, and annual household income (RMB) were negatively correlated with negative psychological status (P < 0.01). Residents' satisfaction with standardized training mode plays a complete mediating role between annual household income and negative psychological status. Conclusions: Our findings emphasize the importance of concentrating on resident's psychological status and family economic situation. And relative departments should take action to optimize the standardized training mode to improve the satisfaction.

Hydrogels With Tunable Mechanical Properties Based on Photocleavable Proteins.

Hydrogels with photo-responsive mechanical properties have found broad biomedical applications, including delivering bioactive molecules, cell culture, biosensing, and tissue engineering. Here, using a photocleavable protein, PhoCl, as the crosslinker we engineer two types of poly(ethylene glycol) hydrogels whose mechanical stability can be weakened or strengthened, respectively, upon visible light illumination. In the photo weakening hydrogels, photocleavage leads to rupture of the protein crosslinkers, and decrease of the mechanical properties of the hydrogels. In contrast, in the photo strengthening hydrogels, by properly choosing the crosslinking positions, photocleavage does not rupture the crosslinking sites but exposes additional cryptical reactive cysteine residues. When reacting with extra maleimide groups in the hydrogel network, the mechanical properties of the hydrogels can be enhanced upon light illumination. Our study indicates that photocleavable proteins could provide more designing possibilities than the small-molecule counterparts. A proof-of-principle demonstration of spatially controlling the mechanical properties of hydrogels was also provided.

Dihydroartemisinin Sensitizes Esophageal Squamous Cell Carcinoma to Cisplatin by Inhibiting Sonic Hedgehog Signaling.

Platinum-based regimens have been routinely used in the clinical treatment of patients with esophageal squamous cell carcinoma (ESCC). However, administration of these drugs is frequently accompanied by drug resistance. Revealing the underlying mechanisms of the drug resistance and developing agents that enhance the sensitivity to platinum may provide new therapeutic strategies for the patients. In the present study, we found that the poor outcome of ESCC patients receiving platinum-based regimens was associated with co-expression of Shh and Sox2. The sensitivity of ESCC cell lines to cisplatin was related to their activity of Shh signaling. Manipulating of Shh expression markedly changed the sensitivity of ESCC cells to platinum. Continuous treatment with cisplatin resulted in the activation of Shh signaling and enhanced cancer stem cell-like phenotypes in ESCC cells. Dihydroartemisinin (DHA), a classic antimalarial drug, was identified as a novel inhibitor of Shh pathway. Treatment with DHA attenuated the cisplatin-induced activation of the Shh pathway in ESCC cells and synergized the inhibitory effect of cisplatin on proliferation, sphere and colony formation of ALDH-positive ESCC cells in vitro and growth of ESCC cell-derived xenograft tumors in vivo. Taken together, these results demonstrate that the Shh pathway is an important player in cisplatin-resistant ESCC and DHA acts as a promising therapeutic agent to sensitize ESCC to cisplatin treatment.

Betulinic Acid Inhibits Endometriosis Through Suppression of Estrogen Receptor ß Signaling Pathway.

Endometriosis is an inflammatory gynecological disorder characterized by endometrial tissue growth located outside of the uterine cavity in addition to chronic pelvic pain and infertility. In this study, we aim to develop a potential therapeutic treatment based on the pathogenesis and mechanism of Endometriosis. Our preliminary data showed that the expression of estrogen receptor ß (ERß) was significantly increased, while ERα was significantly decreased, in endometriotic cells compared to normal endometrial cells. Further investigation showed that betulinic acid (BA) treatment suppressed ERß expression through epigenetic modification on the ERß promoter, while had no effect on ERα expression. In addition, BA treatment suppresses ERß target genes, including superoxide dismutase 2 (SOD2), nuclear respiratory factor-1 (NRF1), cyclooxygenase 2 (COX2), and matrix metalloproteinase-1 (MMP1), subsequently increasing oxidative stress, triggering mitochondrial dysfunction, decreasing elevated proinflammatory cytokines, and eventually suppressing endometriotic cell proliferation, mimicking the effect of ERß knockdown. On the other hand, gain of ERß by lentivirus infection in normal endometrial cells resulted in increased cell proliferation and proinflammatory cytokine release, while BA treatment diminished this effect through ERß suppression with subsequent oxidative stress and apoptosis. Our results indicate that ERß may be a major driving force for the development of endometriosis, while BA inhibits Endometriosis through specific suppression of the ERß signaling pathway. This study provides a novel therapeutic strategy for endometriosis treatment through BA-mediated ERß suppression.

Berberine Ameliorates Prenatal Dihydrotestosterone Exposure-Induced Autism-Like Behavior by Suppression of Androgen Receptor.

Many epidemiology studies have shown that maternal polycystic ovary syndrome (PCOS) results in a greater risk of autism spectrum disorders (ASD) development, although the detailed mechanism remains unclear. In this study, we aimed to investigate the potential mechanism and provide a possible treatment for PCOS-mediated ASD through three experiments: Experiment 1: real-time PCR and western blots were employed to measure gene expression in human neurons, and the luciferase reporter assay and chromatin immunoprecipitation (ChIP) was used to map the responsive elements on related gene promoters. Experiment 2: pregnant dams were prenatally exposed to dihydrotestosterone (DHT), androgen receptor (AR) knockdown (shAR) in the amygdala, or berberine (BBR), and the subsequent male offspring were used for autism-like behavior (ALB) assay followed by biomedical analysis, including gene expression, oxidative stress, and mitochondrial function. Experiment 3: the male offspring from prenatal DHT exposed dams were postnatally treated by either shAR or BBR, and the offspring were used for ALB assay followed by biomedical analysis. Our findings showed that DHT treatment suppresses the expression of estrogen receptor ß (ERß) and superoxide dismutase 2 (SOD2) through AR-mediated hypermethylation on the ERß promoter, and BBR treatment suppresses AR expression through hypermethylation on the AR promoter. Prenatal DHT treatment induces ERß suppression, oxidative stress and mitochondria dysfunction in the amygdala with subsequent ALB behavior in male offspring, and AR knockdown partly diminishes this effect. Furthermore, both prenatal and postnatal treatment of BBR partly restores prenatal DHT exposure-mediated ALB. In conclusion, DHT suppresses ERß expression through the AR signaling pathway by hypermethylation on the ERß promoter, and BBR restores this effect through AR suppression. Prenatal DHT exposure induces ALB in offspring through AR-mediated ERß suppression, and both prenatal and postnatal treatment of BBR ameliorates this effect. We conclude that BBR ameliorates prenatal DHT exposure-induced ALB through AR suppression, this study may help elucidate the potential mechanism and identify a potential treatment through using BBR for PCOS-mediated ASD.