The effect of TFAP2A/ANXA8 axis on ferroptosis of cervical squamous cell carcinoma (CESC) in vitro.

Potential role and associated mechanisms of Annexin A8 (ANXA8), a member of the Annexins family, in cervical squamous cell carcinoma (CESC) are still unclear, despite being upregulated in various malignant tumors. Here, we observed a notably elevated expression of ANXA8 in CESC cells. The inhibition of ANXA8 amplified the susceptibility of CESC cells to Erastin and sorafenib-induced ferroptosis, whereas it exerted minimal influence on DPI7 and DPI10-induced ferroptosis. The results from the Fe2+ concentration assay showed no significant correlation between ANXA8 gene knockdown and intracellular Fe2+ concentration induced by ferroptosis inducers. Western blot analysis demonstrated that the knockdown of ANXA8 did not alter ACSL4 and LPCAT levels under ferroptosis-inducing conditions, but it did result in a reduction in intracellular GSH levels induced by the ferroptosis inducer. Subsequently, we identified TFAP2A as an upstream transcription factor of ANXA8, which plays a role in regulating cell ferroptosis. The knockdown of TFAP2A significantly elevated MDA levels and depressed GSH levels in the presence of a ferroptosis inducer, thereby inhibiting cell ferroptosis. However, this inhibitory effect could be reversed by ANXA8 overexpression. Therefore, our research suggests that the TFAP2A/ANXA8 axis exerts regulatory control over ferroptosis in CESC cells by mediating GSH synthesis in System Xc.

Identification and validation of shared genes and key pathways in endometriosis and endometriosis-associated ovarian cancer by weighted gene co-expression network analysis and machine learning algorithms.

BACKGROUND: Epidemiological studies reported that patients with endometriosis had an increased risk of developing endometriosis-associated ovarian cancer (EAOC). The present study aimed to identify shared genes and key pathways that commonly interacted between EAOC and endometriosis. METHODS: The expression matrix of ovarian cancer and endometriosis were collected from the Gene Expression Omnibus database. The weighted gene co-expression network analysis (WGCNA) was used to construct co-expression gene network. Machine learning algorithms were applied to identify characteristic genes. CIBERSORT deconvolution algorithm was used to explore the difference in tumor immune microenvironment. Furthermore, diagnostic nomogram was constructed and evaluated for supporting clinical practicality. RESULTS: We identified 262 shared genes between EAOC and endometriosis via WGCNA analysis. They were mainly enriched in cytokine-cytokine receptor interaction. After protein-protein interaction network and machine learning algorithms, we recognized two characteristic genes (EDNRA, OCLN) and established a nomogram that presented an outstanding predictive performance. The hub genes demonstrated remarkable associations with immunological functions. Survival analysis indicated that dysregulated expressions of EDNRA and OCLN were closely correlated with prognosis of ovarian cancer patients. gene set enrichment analyses revealed that the two characteristic genes were mainly enriched in the cancer- and immune-related pathways. CONCLUSION: Our findings pave the way for further investigation of potential candidate genes and will aid in improving the diagnosis and treatment of EAOC in endometriosis patients. More research is required to determine the exact mechanisms by which these two hub genes affecting the development and progression of EAOC from endometriosis.

Identifying Immune Cell Infiltration and Hub Genes Related to M2 Macrophages in Endometriosis by Bioinformatics Analysis.

Endometriosis (EM) is a chronic, estrogen-dependent inflammatory disease. Presently, the pathophysiology of EM is still unclear, and numerous studies have established that the immune system plays a major role in the pathophysiology of EM. Six microarray datasets were downloaded from the GEO public database. A total of 151 endometrial samples (72 ectopic endometria and 79 controls) were included in this study. CIBERSORT and ssGSEA were applied to calculate the immune infiltration of EM and control samples. Moreover, we validated four different correlation analyses to explore immune microenvironment of EM and finally identified M2 macrophage-related hub genes and further conducted the specific immunologic signaling pathway analysis by GSEA. The logistic regression model was investigated by ROC and further validated by two external datasets. From the results of the two immune infiltration assays, we concluded that M2 macrophages, regulatory T cells (Tregs), M1 macrophages, activated B cells, T follicular helper cells, activated dendritic cells, and resting NK cells have a significant difference between control and EM tissues. Through multidimensional correlation analysis, we found that macrophages play an important central role in cell-to-cell interactions, especially M2 macrophages. Four immune-related hub genes, namely FN1, CCL2, ESR1, and OCLN, are closely related to M2 macrophages and play a crucial role in the occurrence and immune microenvironment of endometriosis. The combined AUC of ROC prediction model in test and validation sets were 0.9815 and 0.8206, respectively. We conclude that M2 macrophages play a central role in the immune-infiltrating microenvironment of EM.

Histone modifications regulate pioneer transcription factor cooperativity.

Pioneer transcription factors have the ability to access DNA in compacted chromatin1. Multiple transcription factors can bind together to a regulatory element in a cooperative way, and cooperation between the pioneer transcription factors OCT4 (also known as POU5F1) and SOX2 is important for pluripotency and reprogramming2-4. However, the molecular mechanisms by which pioneer transcription factors function and cooperate on chromatin remain unclear. Here we present cryo-electron microscopy structures of human OCT4 bound to a nucleosome containing human LIN28B or nMATN1 DNA sequences, both of which bear multiple binding sites for OCT4. Our structural and biochemistry data reveal that binding of OCT4 induces changes to the nucleosome structure, repositions the nucleosomal DNA and facilitates cooperative binding of additional OCT4 and of SOX2 to their internal binding sites. The flexible activation domain of OCT4 contacts the N-terminal tail of histone H4, altering its conformation and thus promoting chromatin decompaction. Moreover, the DNA-binding domain of OCT4 engages with the N-terminal tail of histone H3, and post-translational modifications at H3K27 modulate DNA positioning and affect transcription factor cooperativity. Thus, our findings suggest that the epigenetic landscape could regulate OCT4 activity to ensure proper cell programming.

Applications and prospects of cryo-EM in drug discovery.

Drug discovery is a crucial part of human healthcare and has dramatically benefited human lifespan and life quality in recent centuries, however, it is usually time- and effort-consuming. Structural biology has been demonstrated as a powerful tool to accelerate drug development. Among different techniques, cryo-electron microscopy (cryo-EM) is emerging as the mainstream of structure determination of biomacromolecules in the past decade and has received increasing attention from the pharmaceutical industry. Although cryo-EM still has limitations in resolution, speed and throughput, a growing number of innovative drugs are being developed with the help of cryo-EM. Here, we aim to provide an overview of how cryo-EM techniques are applied to facilitate drug discovery. The development and typical workflow of cryo-EM technique will be briefly introduced, followed by its specific applications in structure-based drug design, fragment-based drug discovery, proteolysis targeting chimeras, antibody drug development and drug repurposing. Besides cryo-EM, drug discovery innovation usually involves other state-of-the-art techniques such as artificial intelligence (AI), which is increasingly active in diverse areas. The combination of cryo-EM and AI provides an opportunity to minimize limitations of cryo-EM such as automation, throughput and interpretation of medium-resolution maps, and tends to be the new direction of future development of cryo-EM. The rapid development of cryo-EM will make it as an indispensable part of modern drug discovery.

Histone modifications regulate pioneer transcription factor binding and cooperativity.

Pioneer transcription factors have the ability to access DNA in compacted chromatin. Multiple transcription factors can bind together to a regulatory element in a cooperative way and cooperation between pioneer transcription factors Oct4 and Sox2 is important for pluripotency and reprogramming. However, the molecular mechanisms by which pioneer transcription factors function and cooperate remain unclear. Here we present cryo-EM structures of human Oct4 bound to a nucleosome containing human Lin28B and nMatn1 DNA sequences, which bear multiple binding sites for Oct4. Our structural and biochemistry data reveal that Oct4 binding induces changes to the nucleosome structure, repositions the nucleosomal DNA and facilitates cooperative binding of additional Oct4 and of Sox2 to their internal binding sites. The flexible activation domain of Oct4 contacts the histone H4 N-terminal tail, altering its conformation and thus promoting chromatin decompaction. Moreover, the DNA binding domain of Oct4 engages with histone H3 N-terminal tail, and posttranslational modifications at H3K27 modulate DNA positioning and affect transcription factor cooperativity. Thus, our data show that the epigenetic landscape can regulate Oct4 activity to ensure proper cell reprogramming.

Non-canonical bivalent H3K4me3K9me3 recognition by Spindlin1/C11orf84 complex.

Bivalent chromatin with active H3K4me3 and repressive H3K27me3 was initially identified in embryonic stem cells (ESCs) to poise expression of developmental genes upon lineage commitment. Since then, many more different bivalent modifications have been demonstrated in both ESCs and fully differentiated cells. Bivalency not only spatiotemporally controls gene transcription but also acts to fine-tune the level of transcription during development. Although increasing number of studies demonstrated the functional significance of bivalent chromatin, the molecular connection of bivalent chromatin and transcriptional regulation remains largely elusive. Recently, we showed Spindlin1/C11orf84 complex prefers to recognize the non-canonical histone H3K4me3K9me3 bivalent mark, which is required for timely ribosomal RNA transcription. Here, we hypothesize the recognition of K4me3 and K9me3 at the same histone tail by Spindlin1/C11orf84 complex may serve as a general mechanism of conversion from a repressed to an active chromatin structure for transcriptional activation.

Withaferin A inhibits proliferation of human endometrial cancer cells via transforming growth factor-ß (TGF-ß) signalling.

The present study was designed to evaluate the anticancer effects of withaferin A against the human endometrial cancer via modulation of transforming growth factor-ß (TGF-ß) signalling. The results of the present study revealed that withaferin A exerts a dose and time-dependent antiproliferative effects against the human KLE endometrial cancer cells with comparatively lower toxicity against the THESCs normal cells. The IC50 of withaferin A against the KLE endometrial cancer cells was found to 10 µM. The results showed that withaferin A induced apoptosis and G2/M cell cycle arrest of the KLE cells which was associated with alteration of the apoptosis and cell cycle related proteins. In addition, the transwell assays showed that the migration and invasion of the KLE cells were inhibited by 53 and 40%, respectively. Finally, the effects of withaferin A were also examined on the TGF-ß signalling pathway. The results showed that withaferin A blocked TGF-ß-dependent Smad2 phosphorylation and expression of other TGF-ß-related proteins in KLE cells. Summing up, the results suggest that withaferin A inhibits the proliferation of the human endometrial carcinoma via TGF-ß signalling.

Structural mechanism of bivalent histone H3K4me3K9me3 recognition by the Spindlin1/C11orf84 complex in rRNA transcription activation.

Spindlin1 is a unique multivalent epigenetic reader that facilitates ribosomal RNA transcription. In this study, we provide molecular and structural basis by which Spindlin1 acts in complex with C11orf84 to preferentially recognize non-canonical bivalent mark of trimethylated lysine 4 and lysine 9 present on the same histone H3 tail (H3K4me3K9me3). We demonstrate that C11orf84 binding stabilizes Spindlin1 and enhances its association with bivalent H3K4me3K9me3 mark. The functional analysis suggests that Spindlin1/C11orf84 complex can displace HP1 proteins from H3K4me3K9me3-enriched rDNA loci, thereby facilitating the conversion of these poised rDNA repeats from the repressed state to the active conformation, and the consequent recruitment of RNA Polymerase I for rRNA transcription. Our study uncovers a previously unappreciated mechanism of bivalent H3K4me3K9me3 recognition by Spindlin1/C11orf84 complex required for activation of rRNA transcription.

Complete salpingectomy versus tubal ligation during cesarean section: a systematic review and meta-analysis.

OBJECTIVE: This study aimed to evaluate the safety profile and feasibility of complete salpingectomy during cesarean delivery in women desiring permanent sterilization. DATA SOURCES: We searched MEDLINE, Embase, and Cochrane Central Register of Controlled Trials from inception to July 2018. Study Selection: studies comparing total salpingectomy with tubal ligation during cesarean deliveries were included. RESULTS: Nine studies involving a total of 1274 participants were eligible. Our analyses showed that the total operative time was slightly longer for the bilateral salpingectomy than for the tubal ligation group (MD = 5.81, 95% CI: 0.85-10.77). Two comparison groups were comparable with regard to intraoperative complications (RR = 1.42, 95% CI: 0.65-3.11), postoperative complications (RR = 1.70, 95% CI: 0.83-3.48), estimated blood loss in total procedures, need for blood transfusion, operative complications, risk of postpartum hemorrhage, surgical site infection, ICU admission, need for presentation to hospital, short-term ovarian reserve, and completion rate of sterilization surgeries (RR = 0.90, 95% CI: 0.80-1.00). CONCLUSION: Complete salpingectomy slightly prolonged surgical time by a reasonable range and showed similar safety profile and greater cost-effectiveness than tubal ligation as permanent sterilization procedures at cesarean delivery.

Removal, seasonal variation, and environmental impact of parabens in a municipal wastewater treatment facility in Guangzhou, China.

The occurrence, seasonal variation, and environmental impact of five widely used parabens, methyl-(MeP), ethyl-(EtP), n-propyl-(n-PrP), n-butyl-(n-BuP), and benzyl-(BzP) parabens, were investigated in a municipal wastewater treatment plant (WWTP) located in Guangzhou, China, for 1 year. The concentrations of ∑5parabens in the influent and the effluent were 94.2-957 and 0.89-14.7 ng L-1, respectively. The influent paraben concentrations in autumn were significantly lower than in winter, spring, and summer, and the concentrations were generally higher in spring. The removal efficiencies of ∑5parabens in the dissolved phase were over 96%, with high efficiencies in MeP, EtP, and n-PrP. Risk assessment indicated that parabens in the effluent were not likely to pose an environmental risk to aquatic ecosystems. The present study indicates that the treatment processes employed in full-scale WWTPs are effective at removing parabens and highlights the possibility of utilizing WWTPs for restoring water quality in riverine and coastal regions heavily impacted by paraben contamination.

Occurrence, composition and biological risk of organophosphate esters (OPEs) in water of the Pearl River Estuary, South China.

Since the production of brominated flame retardants has been gradually phased out, organophosphate esters (OPEs) are increasingly used as the substitutes. Given their toxicity and water solubility, OPEs may jeopardize the aquatic environment and organisms. Here, we examined the concentration, composition, and biological risk of OPEs in the water collected from the eight major waterways in the Pearl River Delta, a highly industrialized region in China. We found a widespread occurrence of OPEs in this region (∑9OPEs: 134 to 442 ng L-1), dominated by TCPP, TCEP, and TnBP. Halogenated OPEs were dominant over alkyl and aromatic OPEs. The biological risk of OPEs, mainly contributed by TPhP and TnBP, was low (RQ < 0.1). The contamination level of OPEs in the Pearl River Delta was likely associated with the degree of industrial activities. Although OPEs posed low risk to aquatic organisms, more attention should be paid to some OPEs in the future, such as TnBP, due to the high usage and toxicity. Considering the concentrations of OPEs worldwide and their usage, OPEs may become the emerging pollutants of global concern in the next decade.

Occurrence, distribution, and potential risks of environmental corticosteroids in surface waters from the Pearl River Delta, South China.

The occurrence, spatiotemporal distribution, and potential risks of 21 glucocorticoids (GCs) and 3 mineralocorticoids (MCs) in four rivers were studied by investigating the surface waters from the Pearl River Delta (PRD), South China. These environmental corticosteroids (ECs) were commonly present in the river surface waters with average concentrations varying from <0.17 ng/L for fluticasone propionate to 5.6 ng/L for clobetasone butyrate; and cortisone had the highest concentration, 32.9 ng/L. The total ECs ranged in concentration from undetectable to 83.3 ng/L, with a mean and median of 8.1 ng/L and 4.8 ng/L, respectively. Spatially the total EC concentration levels in the Pearl River system occurred in the following order: Zhujiang River (ZR) > Dongjiang River (DR) > Shiziyang waterway (SW) > Beijiang River (BR). These levels generally demonstrated a trend of increasing from upstream to midstream or downstream then attenuating toward the estuary. Considerable seasonal variations in the ECs differed among rivers. Higher ECs concentrations in winter were mostly found in the ZR, whereas lower levels were found in the DR. Moreover, the temporal variations of the ECs were marginal in the BR and SW. These spatiotemporal distributions of the ECs might have been simultaneously influenced by pollution sources derived from anthropogenic activities and river hydrologic conditions. Correlation analyses indicated that dissolved organic carbon (DOC) could play a key role in the occurrence and distribution of ECs in an aquatic environment. Risk assessment demonstrated that the occurrence of ECs might have posed medium to high risk to aquatic organisms in the Pearl River.

Vertical profile of soil/sediment pollution and microbial community change by e-waste recycling operation.

The present study aims to assess the effect of electronic waste (e-waste) recycling on microbial community and the underlying modulation mechanism. Core soil/sediment samples were collected from an abandoned e-waste burning site and neighboring farmland/stream sites in Guiyu, China. High concentrations and health risks of toxic heavy metals, particularly, Sb and Sn, and halogenated flame retardants (HFRs), including decabromodiphenyl ether (BDE 209) and decabromodiphenyl ethane (DBDPE) were mostly retained at the top surface layers of soils/sediments (0-30cm) after more than one year of natural vertical diffusion and microbe-facilitated biodegradation. Heavy metals, such as Ag, Cd, Cu, Pb, Sb, and Sn, played a critical role for the reduction of microbial diversity. This is the first study reporting the open burning of e-waste caused an obvious heat effect and enriched thermophilic/mesophilic microbes in local area. The acid washing during e-waste recycling process may result in the enrichment of acidophilic microbes. This investigation showed that e-waste processing operation resulted in not only severe pollution of the soils/sediments by various pollutants, but also reduction of microbial diversity that was difficult to self-store by the local ecosystem.

Trace metals in e-waste lead to serious health risk through consumption of rice growing near an abandoned e-waste recycling site: Comparisons with PBDEs and AHFRs.

Despite the endeavour to eradicate informal e-waste recycling, remediation of polluted sites is not mandatory in many developing countries and thus the hazard of pollutants remaining in soil is often overlooked. It is noteworthy that a majority of previous studies only analysed a few pollutants in e-waste to reflect the impact of informal e-waste recycling. However, the actual impact may have been largely underestimated since e-waste contains various groups of pollutants and the effect of some emerging pollutants in e-waste remains unexplored. Thus, this study examined the contamination of metals, PBDEs and AHFRs in the vicinity of an abandoned e-waste recycling site. The accumulation and translocation of these pollutants in rice plants cultivated at the nearby paddy field were measured to estimate the health risk through rice consumption. We revealed that the former e-waste burning site was still seriously contaminated with some metals (e.g. Sn, Sb and Ag, Igeo > 5), PBDEs (Igeo > 3) and AHFRs (Igeo > 3), which can disperse to the nearby paddy field and stream. The rice plants can effectively absorb some metals (e.g. Mo, Cr and Mn, BCF > 1), but not PBDEs and AHFRs (BCF < 0.15), from soil and translocate them to the leaves. Alarmingly, the health risk through rice consumption was high primarily due to Sb and Sn (HQ > 20), whereas PBDEs and AHFRs had limited contribution (HQ < 0.08). Our results imply that abandoned e-waste recycling sites still act as the pollution source, jeopardising the surrounding environment and human health. Since some trace metals (e.g. Sb and Sn) are seldom monitored, the impact of informal e-waste recycling would be more notorious than previously thought. Remediation work should be conducted promptly in abandoned e-waste recycling sites to protect the environment and human health.

Matrix metalloproteinase-7 may serve as a novel biomarker for cervical cancer.

BACKGROUND: The biological and clinical significance of matrix metalloproteinase-7 (MMP-7) in cervical cancer remains unknown. Here, we investigated the function of MMP-7 in cervical cancer cells and evaluated its clinical significance in both tissues and serum from cervical cancer patients. METHODS: First, we analyzed the expression of MMP-7 in cervical cancer using Oncomine microarray data and examined its expression in cervical tissues by quantitative real-time polymerase chain reaction and Western blotting. Second, we utilized gene silencing to explore the role of MMP-7 in cells. Finally, we examined the MMP-7 levels in patients with cervical cancer and normal serum by enzyme-linked immunosorbent assay. Moreover, we further investigated the relationship between MMP-7 expression and pathological features. RESULTS: The mRNA and protein MMP-7 levels were higher in cervical cancer tissues than in healthy controls. Silencing of MMP-7 significantly decreased cervical cancer cell proliferation, migration, and invasion. The serum MMP-7 levels were significantly higher in cervical cancer patients than in healthy subjects (P<0.01). Further, higher MMP-7 expression was associated with increased lymph metastasis (P=0.021), pathological grade (P=0.039, P=0.047), and clinical stage (P=0.049, P=0.046). CONCLUSION: MMP-7 appears to act as an oncogene in cervical cancer cells and is involved in cell proliferation, migration, and invasion. MMP-7 expression was significantly higher in the tissue and serum of cervical cancer patients compared to healthy individuals and was correlated with increased pathalogical grade, clinical stage, and lymph metastasis. Therefore, our data provide novel evidence that MMP-7 may be a clinically relevant biomarker for cervical cancer.

Structural and mechanistic insights into UHRF1-mediated DNMT1 activation in the maintenance DNA methylation.

UHRF1 plays multiple roles in regulating DNMT1-mediated DNA methylation maintenance during DNA replication. The UHRF1 C-terminal RING finger functions as an ubiquitin E3 ligase to establish histone H3 ubiquitination at Lys18 and/or Lys23, which is subsequently recognized by DNMT1 to promote its localization onto replication foci. Here, we present the crystal structure of DNMT1 RFTS domain in complex with ubiquitin and highlight a unique ubiquitin binding mode for the RFTS domain. We provide evidence that UHRF1 N-terminal ubiquitin-like domain (UBL) also binds directly to DNMT1. Despite sharing a high degree of structural similarity, UHRF1 UBL and ubiquitin bind to DNMT1 in a very distinct fashion and exert different impacts on DNMT1 enzymatic activity. We further show that the UHRF1 UBL-mediated interaction between UHRF1 and DNMT1, and the binding of DNMT1 to ubiquitinated histone H3 that is catalyzed by UHRF1 RING domain are critical for the proper subnuclear localization of DNMT1 and maintenance of DNA methylation. Collectively, our study adds another layer of complexity to the regulatory mechanism of DNMT1 activation by UHRF1 and supports that individual domains of UHRF1 participate and act in concert to maintain DNA methylation patterns.

Non-structural protein 1 of H3N2 influenza A virus induces nucleolar stress via interaction with nucleolin.

The nucleolus is a stress sensor associated with cell cycle progression and a central hub for the replication of pathogenic RNA viruses. However, the role of nucleolus in influenza A virus infection has not been well studied. Here we show that the interaction between NS1 protein of influenza A/Shantou/602/06 (H3N2) and nucleolin, a ubiquitous protein of nucleolus repressed RNA Pol I-dependent transcription via establishing hyper-methylation in the UCE of rRNA gene promoter. NS1 expressed cells showed significant association of ribosomal proteins with MDM2, and p53 accumulation, suggesting induced nucleolar stress. Disruption of the interaction of NS1 with nucleolin or overexpression of nucleolin in NS1 expressed cells revived RNA Pol I-dependent transcription, indicating nucleolin could be one target for NS1 to repress rRNA synthesis of host cells. Our present study suggests that NS1 protein of H3N2 could induce nucleolar stress based on epigenetic alteration of rRNA gene promoter via interaction with nucleolin.