Genome-wide sequencing identified extrachromosomal circular DNA as a transcription factor-binding motif of the senescence genes that govern replicative senescence in human mesenchymal stem cells.

Introduction: Mesenchymal stem cells (MSCs) have long been postulated as an important source cell in regenerative medicine. During subculture expansion, mesenchymal stem cell (MSC) senescence diminishes their multi-differentiation capabilities, leading to a loss of therapeutic potential. Up to date, the extrachromosomal circular DNAs (eccDNAs) have been demonstrated to be involved in senescence but the roles of eccDNAs during MSC. Methods: Here we explored eccDNA profiles in human bone marrow MSCs (BM-MSCs). EccDNA and mRNA was purified and sequenced, followed by quantification and functional annotation. Moreover, we mapped our datasets with the downloading enhancer and transcription factor-regulated genes to explore the potential role of eccDNAs. Results: Sequentially, gene annotation analysis revealed that the majority of eccDNA were mapped in the intron regions with limited BM-MSC enhancer overlaps. We discovered that these eccDNA motifs in senescent BMSCs acted as motifs for binding transcription factors (TFs) of senescence-related genes. Discussion: These findings are highly significant for identifying biomarkers of senescence and therapeutic targets in mesenchymal stem cells (MSCs) for future clinical applications. The potential of eccDNA as a stable therapeutic target for senescence-related disorders warrants further investigation, particularly exploring chemically synthesized eccDNAs as transcription factor regulatory elements to reverse cellular senescence.

Regulation of Hippo/YAP axis in colon cancer progression by the deubiquitinase JOSD1.

Colon cancer is a prevalent malignancy, while recent studies revealed the dys-regulation of Hippo signaling as the important driver for colon cancer progression. Several studies have indicated that post-translational modifications on YAP play crucial roles in both Hippo signaling activity and cancer progression. This raises a puzzling question about why YAP/TAZ, an auto-inhibitory pathway, is frequently over-activated in colon cancer, despite the suppressive cascade of Hippo signaling remaining operational. The protein stability of YAP is subject to a tiny balance between ubiquitination and deubiquitination processes. Through correlation analysis of DUBs (deubiquitinases) expression and Hippo target gene signature in colon cancer samples, we found JOSD1 as a critical deubiquitinase for Hippo signaling and colon cancer progression. JOSD1 could facilitate colon cancer progression and in colon cancer, inhibition of JOSD1 via shRNA has been demonstrated to impede tumorigenesis. Furthermore, molecular mechanism studies have elucidated that JOSD1 enhances the formation of the Hippo/YAP transcriptome by impeding K48-linked polyubiquitination on YAP. ChIP assays have shown that YAP binds to JOSD1's promoter region, promoting its gene transcription. These results suggest that JOSD1 is involved in both activating and being targeted by the Hippo signaling pathway in colon cancer. Consequently, a positive regulatory loop between JOSD1 and Hippo signaling has been identified, underscoring their interdependence during colon cancer progression. Thus, targeting JOSD1 may represent a promising therapeutic approach for managing colon cancer.

Elevating Detection Performance in Optical Remote Sensing Image Object Detection: A Dual Strategy with Spatially Adaptive Angle-Aware Networks and Edge-Aware Skewed Bounding Box Loss Function.

In optical remote sensing image object detection, discontinuous boundaries often limit detection accuracy, particularly at high Intersection over Union (IoU) thresholds. This paper addresses this issue by proposing the Spatial Adaptive Angle-Aware (SA3) Network. The SA3 Network employs a hierarchical refinement approach, consisting of coarse regression, fine regression, and precise tuning, to optimize the angle parameters of rotated bounding boxes. It adapts to specific task scenarios using either class-aware or class-agnostic strategies. Experimental results demonstrate its effectiveness in significantly improving detection accuracy at high IoU thresholds. Additionally, we introduce a Gaussian transform-based IoU factor during angle regression loss calculation, leading to the development of Edge-aware Skewed Bounding Box Loss (EAS Loss). The EAS loss enhances the loss gradient at the final stage of angle regression for bounding boxes, addressing the challenge of further learning when the predicted box angle closely aligns with the real target box angle. This results in increased training efficiency and better alignment between training and evaluation metrics. Experimental results show that the proposed method substantially enhances the detection accuracy of ReDet and ReBiDet models. The SA3 Network and EAS loss not only elevate the mAP of the ReBiDet model on DOTA-v1.5 to 78.85% but also effectively improve the model's mAP under high IoU threshold conditions.

USP36 promotes tumorigenesis and tamoxifen resistance in breast cancer by deubiquitinating and stabilizing ERα.

BACKGROUND: Breast cancer is the most prevalent cancer in women globally. Over-activated estrogen receptor (ER) α signaling is considered the main factor in luminal breast cancers, which can be effectively managed with selective estrogen receptor modulators (SERMs) like tamoxifen. However, approximately 30-40% of ER + breast cancer cases are recurrent after tamoxifen therapy. This implies that the treatment of breast cancer is still hindered by resistance to tamoxifen. Recent studies have suggested that post-translational modifications of ERα play a significant role in endocrine resistance. The stability of both ERα protein and its transcriptome is regulated by a balance between E3 ubiquitin ligases and deubiquitinases. According to the current knowledge, approximately 100 deubiquitinases are encoded in the human genome, but it remains unclear which deubiquitinases play a critical role in estrogen signaling and endocrine resistance. Thus, decoding the key deubiquitinases that significantly impact estrogen signaling, including the control of ERα expression and stability, is critical for the improvement of breast cancer therapeutics. METHODS: We used several ER positive breast cancer cell lines, DUB siRNA library screening, xenograft models, endocrine-resistant (ERα-Y537S) model and performed immunoblotting, real time PCR, RNA sequencing, immunofluorescence, and luciferase activity assay to investigate the function of USP36 in breast cancer progression and tamoxifen resistance. RESULTS: In this study, we identify Ubiquitin-specific peptidase 36 (USP36) as a key deubiquitinase involved in ERα signaling and the advancement of breast cancer by deubiquitinases siRNA library screening. In vitro and in vivo studies showed that USP36, but not its catalytically inactive mutant (C131A), could promote breast cancer progression through ERα signaling. Conversely, silencing USP36 inhibited tumorigenesis. In models resistant to endocrine therapy, silencing USP36 destabilized the resistant form of ERα (Y537S) and restored sensitivity to tamoxifen. Molecular studies indicated that USP36 inhibited K48-linked polyubiquitination of ERα and enhanced the ERα transcriptome. It is interesting to note that our results suggest USP36 as a novel biomarker for treatment of breast cancer. CONCLUSION: Our study revealed the possibility that inhibiting USP36 combined with tamoxifen could provide a potential therapy for breast cancer.

YAP represses the TEAD-NF-κB complex and inhibits the growth of clear cell renal cell carcinoma.

The Hippo pathway is generally understood to inhibit tumor growth by phosphorylating the transcriptional cofactor YAP to sequester it to the cytoplasm and reduce the formation of YAP-TEAD transcriptional complexes. Aberrant activation of YAP occurs in various cancers. However, we found a tumor-suppressive function of YAP in clear cell renal cell carcinoma (ccRCC). Using cell cultures, xenografts, and patient-derived explant models, we found that the inhibition of upstream Hippo-pathway kinases MST1 and MST2 or expression of a constitutively active YAP mutant impeded ccRCC proliferation and decreased gene expression mediated by the transcription factor NF-κB. Mechanistically, the NF-κB subunit p65 bound to the transcriptional cofactor TEAD to facilitate NF-κB-target gene expression that promoted cell proliferation. However, by competing for TEAD, YAP disrupted its interaction with NF-κB and prompted the dissociation of p65 from target gene promoters, thereby inhibiting NF-κB transcriptional programs. This cross-talk between the Hippo and NF-κB pathways in ccRCC suggests that targeting the Hippo-YAP axis in an atypical manner-that is, by activating YAP-may be a strategy for slowing tumor growth in patients.

Nanopillar array-based electrochemical aptamer sensor for STX sensitivity detection.

Saxitoxin (STX) is a cyanotoxin with high toxicity, and therefore, there is an urgent need to develop a facile detection method for STX. In this study, an ordered nanopillar array-based electrochemical aptasensor was fabricated for the high-performance detection of STX. The anti-STX aptamer with methylene blue (MB) incorporated at the 3'-end (MB-Apt) was immobilized at the surface of an Au@PAN nanopillar array electrode and used as the recognition element. The proposed aptasensor demonstrated highly sensitive and selective STX detection because of synergistic catalysis effects of MB and ordered nanopillar arrays along with the selection of MB-Apt. The nanopillar array-based electrochemical aptasensor exhibited high sensitivity over a wide linear concentration range of 1 pM-3 nM with a linear regression equation of ΔI (µA) = 28.0 + 6.9 × log[STX] (R2 = 0.98079) and 3-100 nM with a linear regression equation of ΔI (µA) = 10.7 + 43.4 × log[STX] (R2 = 0.98772), where R is the correlation coefficient. In addition, the limit of detection (LOD) was as low as 1 pM. Furthermore, the designed aptasensor demonstrated excellent selectivity toward STX, preventing interference from neo-STX, okadaic acid, and common metal ions. The presented orderly nanopillar array-based strategy to develop an electrochemical aptasensor for STX detection offers a promising method for developing high-performance electrochemical sensors, and the presented aptasensor should find useful application in the detection of shellfish poison.

Efficacy of surgical treatment of perianal infection in patients with hematological malignancy.

The efficacy of surgical intervention for perianal infection in patients with hematologic malignancies is not well established. Therefore, our study aimed to investigate the clinical efficacy and complications of surgical treatment of perianal infection in patients with hematologic malignancies. This retrospective study included patients with hematological malignancies who were diagnosed with perianal infections and treated at the China Aerospace Science & Industry Corporation 731 Hospital between 2018 and 2022. Patient characteristics, hematological data, surgical intervention, and complications, including recurrence and mortality, were analyzed. This study included 156 patients with leukemia aged 2 months to 71 years who were treated surgically for perianal infection, comprising 94 males and 62 females. Perianal infection included 36 cases of abscesses, 91 anal fistulas, and 29 anal fissures accompanied by infection. A total of 36 patients developed severe complications postoperatively, including 4 patients who died, 6 patients with severe incision bleeding, 18 patients with severe pain, 6 patients with sepsis, 12 patients who needed reoperation, 15 patients with hospitalization for more than 2 weeks, and 3 patients with anal stenosis; none of the patients developed anal incontinence. Additionally, risk factors for postoperative complications of perianal infection in patients with hematologic malignancies include leukopenia, agranulocytosis, thrombocytopenia, depth of abscess and not undergone an MRI. Surgical intervention may improve the prognosis of patients with perianal abscess formation, particularly in patients who show no improvement with medical therapy and those who develop perianal sepsis. Granulocytopenia and thrombocytopenia should be improved before surgery, which can significantly reduce postoperative complications. Although these findings are from a case series without a comparator, they may be of value to physicians because to the best of our knowledge, no randomized or prospective studies have been conducted on the management of perianal infections in patients with hematological malignancies.

Mining and exploration of rehabilitation nursing targets for colorectal cancer.

BACKGROUND: There are often subtle early symptoms of colorectal cancer, a common malignancy of the intestinal tract. However, it is not yet clear how MYC and NCAPG2 are involved in colorectal cancer. METHOD: We obtained colorectal cancer datasets GSE32323 and GSE113513 from the Gene Expression Omnibus (GEO). After downloading, we identified differentially expressed genes (DEGs) and performed Weighted Gene Co-expression Network Analysis (WGCNA). We then undertook functional enrichment assay, gene set enrichment assay (GSEA) and immune infiltration assay. Protein-protein interaction (PPI) network construction and analysis were undertaken. Survival analysis and Comparative Toxicogenomics Database (CTD) analysis were conducted. A gene expression heat map was generated. We used TargetScan to identify miRNAs that are regulators of DEGs. RESULTS: 1117 DEGs were identified. Their predominant enrichment in activities like the cellular phase of the cell cycle, in cell proliferation, in nuclear and cytoplasmic localisation and in binding to protein-containing complexes was revealed by Gene Ontology (GO). When the enrichment data from GSE32323 and GSE113513 colon cancer datasets were merged, the primary enriched DEGs were linked to the cell cycle, protein complex, cell cycle control, calcium signalling and P53 signalling pathways. In particular, MYC, MAD2L1, CENPF, UBE2C, NUF2 and NCAPG2 were identified as highly expressed in colorectal cancer samples. Comparative Toxicogenomics Database (CTD) demonstrated that the core genes were implicated in the following processes: colorectal neoplasia, tumour cell transformation, inflammation and necrosis. CONCLUSIONS: High MYC and NCAPG2 expression has been observed in colorectal cancer, and increased MYC and NCAPG2 expression correlates with worse prognosis.

An Efficient Electron Ptychography Method for Retrieving the Object Spectrum from Only a Few Iterations.

We present an efficient approach for electron ptychography based on a mathematical relationship that differs from that underlying the established algorithms of the ptychography iterative engine or the noniterative algorithms like the Wigner-distribution-deconvolution or the single-side-band method. Three variables are handled in this method-the transfer function of the objective lens, the object spectrum, and the diffraction wave whose phase is unknown. In the case of an aberration-corrected electron microscope, one is able to obtain a well-estimated transfer function of the lens. After reducing the number of three variables down to two, we construct an iterative loop between the object spectrum and the diffraction wave, which retrieves the object spectrum within a small number of iterations. We tested this object spectrum retrieval method on both a calculated and an experimental 4D-STEM datasets. By applying this method, we explore the influence of sampling, dose, and the size of illumination aperture on the reconstructed phase images.

A randomized double-blinded trial to assess recurrence of systemic allergic reactions following COVID-19 mRNA vaccination.

BACKGROUND: Systemic allergic reactions (sARs) following coronavirus disease 2019 (COVID-19) mRNA vaccines were initially reported at a higher rate than after traditional vaccines. OBJECTIVE: We aimed to evaluate the safety of revaccination in these individuals and to interrogate mechanisms underlying these reactions. METHODS: In this randomized, double-blinded, phase 2 trial, participants aged 16 to 69 years who previously reported a convincing sAR to their first dose of COVID-19 mRNA vaccine were randomly assigned to receive a second dose of BNT162b2 (Comirnaty) vaccine and placebo on consecutive days in a blinded, 1:1 crossover fashion at the National Institutes of Health. An open-label BNT162b2 booster was offered 5 months later if the second dose did not result in severe sAR. None of the participants received the mRNA-1273 (Spikevax) vaccine during the study. The primary end point was recurrence of sAR following second dose and booster vaccination; exploratory end points included biomarker measurements. RESULTS: Of 111 screened participants, 18 were randomly assigned to receive study interventions. Eight received BNT162b2 second dose followed by placebo; 8 received placebo followed by BNT162b2 second dose; 2 withdrew before receiving any study intervention. All 16 participants received the booster dose. Following second dose and booster vaccination, sARs recurred in 2 participants (12.5%; 95% CI, 1.6 to 38.3). No sAR occurred after placebo. An anaphylaxis mimic, immunization stress-related response (ISRR), occurred more commonly than sARs following both vaccine and placebo and was associated with higher predose anxiety scores, paresthesias, and distinct vital sign and biomarker changes. CONCLUSIONS: Our findings support revaccination of individuals who report sARs to COVID-19 mRNA vaccines. Distinct clinical and laboratory features may distinguish sARs from ISRRs.

Phenotypic switching as a non-genetic mechanism of resistance predicts antibody therapy regimens.

Despite the specificity and effectiveness of antibody therapy, resistance to treatment remains a major barrier for their broad clinical applications. While genetic mutations are known to be critical, the impact of non-genetic mechanisms, such as epigenetic changes and phenotypic adaptations, on resistance to antibody-dependent cellular cytotoxicity (ADCC) is not fully understood. Our study investigated the non-genetic resistance mechanisms that colorectal cancer cells develop against cetuximab and the resulting ADCC pressure. Resistance clones exhibited decreased EGFR/HER2 expressions, enriched interferon-related pathways, and lower NK cell activation. Interestingly, these resistance clones regained sensitivity upon the withdrawal of therapeutic pressure, implying phenotypic plasticity and reversibility. To counter resistance, we developed a mathematical model recapitulating the phenotypic switching dynamics. The model predicted that intermittent dosing strategy outperforms continuous regimen in delaying treatment resistance. Our findings have implications for improving efficacy and circumventing resistance to targeted antibody therapies.

Free PEG Suppresses Anaphylaxis to PEGylated Nanomedicine in Swine.

Covalent conjugation of poly(ethylene glycol) (PEG) is frequently employed to enhance the pharmacokinetics and biodistribution of various protein and nanoparticle therapeutics. Unfortunately, some PEGylated drugs can induce elevated levels of antibodies that can bind PEG, i.e., anti-PEG antibodies (APA), in some patients. APA in turn can reduce the efficacy and increase the risks of allergic reactions, including anaphylaxis. There is currently no intervention available in the clinic that specifically mitigates allergic reactions to PEGylated drugs without the use of broad immunosuppression. We previously showed that infusion of high molecular weight free PEG could safely and effectively suppress the induction of APA in mice and restore prolonged circulation of various PEGylated therapeutics. Here, we explored the effectiveness of free PEG as a prophylaxis against anaphylaxis induced by PEG-specific allergic reactions in swine. Injection of PEG-liposomes (PL) resulted in anaphylactoid shock (pseudoanaphylaxis) within 1-3 min in both naïve and PL-sensitized swine. In contrast, repeated injection of free PEG alone did not result in allergic reactions, and injection of free PEG effectively suppressed allergic reactions to PL, including in previously PL-sensitized swine. These results strongly support the further investigation of free PEG for reducing APA and allergic responses to PEGylated therapeutics.

Development of anti-PEG IgG/IgM/IgE ELISA assays for profiling anti-PEG immunoglobulin response in PEG-sensitized individuals and patients with alpha-gal allergy.

Polyethylene glycol (PEG) is frequently used in various protein and nanomedicine therapeutics. However, various studies have shown that select PEGylated therapeutics can induce production of anti-PEG antibodies (APA), potentially culminating in rapid clearance from the systemic circulation, loss of efficacy and possibly increased risks of allergic reactions. Although IgE is a frequent cause of immediate hypersensitivity reactions (IHR), the role of IgE APA in PEG-related IHR is not well understood, due in part to a lack of standardized assays for measuring IgE APA. Here, we developed a rigorous competitive ELISA method to measure the concentrations of various APA isotypes, including IgE, with picomolar sensitivities. In a small number of serum samples from patients with known PEG allergy, the assay allowed us to detect a strong correlation between IgG and IgE APA in individuals with history of allergic reactions to PEG or PEGylated drugs, but not between IgM and IgE APA. We detected appreciable levels of IgG and IgM APA in individuals with history of alpha-gal allergy, however, they were not elevated relative to those detected in other healthy controls, and we found no pre-existing IgE APA. While preliminary and should be further investigated, these results suggest that differences in the route and mechanism of PEG exposure may drive variability in APA response.

PSMD14 stabilizes estrogen signaling and facilitates breast cancer progression via deubiquitinating ERα.

The over-activation of ERα signaling is regarded as the major driver for luminal breast cancers, which could be effective controlled via selective estrogen receptor modulators (SERM), such as tamoxifen. The endocrine resistance is still a challenge for breast cancer treatment, while recently studies implicate the post-translational modification on ERα play important roles in endocrine resistance. The stability of ERα protein and ERα transcriptome are subject to a balance between E3 ubiquitin ligases and deubiquitinases. Through deubiquitinases siRNA library screening, we discover PSMD14 as a critical deubiquitinase for ERα signaling and breast cancer progression. PSMD14 could facilitate breast cancer progression through ERα signaling in vitro and in vivo, while pharmaceutical inhibition of PSMD14 via Thiolutin could block the tumorigenesis in breast cancer. In endocrine resistant models, PSMD14 inhibition could de-stabilize the resistant form of ERα (Y537S) and restore tamoxifen sensitivity. Molecular studies reveal that PSMD14 could inhibition K48-linked poly-ubiquitination on ERα, facilitate ERα transcriptome. Interestingly, ChIP assay shows that ERα could bind to the promoter region of PSMD14 and facilitate its gene transcription, which indicates PSMD14 is both the upstream modulator and downstream target for ERα signaling in breast cancer. In general, we identified a novel positive feedback loop between PSMD14 and ERα signaling in breast cancer progression, while blockade of PSMD14 could be a plausible strategy for luminal breast cancer.

Intercellular adhesion molecule 1 and selectin l play crucial roles in ulcerative colitis.

Ulcerative colitis (UC) is a chronic inflammatory bowel disease that primarily affects the mucosal layer of the colon (large intestine). However, the relationship between Intercellular Adhesion Molecule-1 (ICAM1), SELL and UC is unclear. The UC datasets, GSE87466 and GSE36807, were downloaded from the gene expression omnibus database. The R package limma was utilized to identify differentially expressed genes (DEGs). Weighted gene co-expression network analysis was conducted. The construction and analysis of protein-protein interaction network, functional enrichment analysis, gene set enrichment analysis, and comparative toxicogenomics database analysis were performed. TargetScan was employed to screen miRNAs regulating central DEGs. Western blot (WB) was used to verify. A total of 2118 DEGs were identified in our study. Gene ontology analysis indicated their enrichment primarily in immune system processes, cellular responses to chemical stimuli, responses to organic substances, responses to external stimuli, and immune responses. Kyoto Encyclopedia of Genes and Genomes analysis revealed that the target cells were mainly enriched in chemokine signaling pathways and TNF signaling pathways. Gene set enrichment analysis enrichment analysis showed significant enrichment in chemokine signaling pathways and cell adhesion molecules. In the Metascape enrichment project, gene ontology terms included regulation of cell activation and positive regulation of immune response. Through the construction and analysis of a protein-protein interaction network, we identified 11 core genes (ICAM1, SELL, CD44, CD40, CCR7, CXCL8, CD19, CCL4, CD274, IL7R, IL1B). We found that the core genes (ICAM1, SELL) were highly expressed in UC samples and lowly expressed in normal samples, suggesting their potential regulatory roles in UC. These core genes were associated with lymphoproliferative disorders, inflammation and necrosis. WB results confirmed the high expression of ICAM1 and SELL in UC. ICAM1 and SELL are highly expressed in UC, and the higher the ICAM1 and SELL genes, the worse the prognosis.

Crack Segmentation Extraction and Parameter Calculation of Asphalt Pavement Based on Image Processing.

Crack disease is one of the most serious and common diseases in road detection. Traditional manual methods for measuring crack detection can no longer meet the needs of road crack detection. In previous work, the authors proposed a crack detection method for asphalt pavements based on an improved YOLOv5s model, which is a better model for detecting various types of cracks in asphalt pavements. However, most of the current research on automatic pavement crack detection is still focused on crack identification and location stages, which contributes little to practical engineering applications. Based on the shortcomings of the above work, and in order to improve its contribution to practical engineering applications, this paper proposes a method for segmenting and analyzing asphalt pavement cracks and identifying parameters based on image processing. The first step is to extract the crack profile through image grayscale, histogram equalization, segmented linear transformation, median filtering, Sauvola binarization, and the connected domain threshold method. Then, the magnification between the pixel area and the actual area of the calibration object is calculated. The second step is to extract the skeleton from the crack profile images of asphalt pavement using the Zhang-Suen thinning algorithm, followed by removing the burrs of the crack skeleton image using the connected domain threshold method. The final step is to calculate physical parameters, such as the actual area, width, segments, and length of the crack with images obtained from the crack profile and skeleton. The results show that (1) the method of local thresholding and connected domain thresholding can completely filter noise regions under the premise of retaining detailed crack region information. (2) The Zhang-Suen iterative refinement algorithm is faster in extracting the crack skeleton of asphalt pavement, retaining the foreground features of the image better, while the connected-domain thresholding method is able to eliminate the missed isolated noise. (3) In comparison to the manual calibration method, the crack parameter calculation method proposed in this paper can better complete the calculation of crack length, width, and area within an allowable margin of error. On the basis of this research, a windowing system for asphalt pavement crack detection, WSPCD1.0, was developed. It integrates the research results from this paper, facilitating automated detection and parameter output for asphalt pavement cracks.

CALM3 affects the prognosis of leukemia and hemorrhoids.

Leukemia is an abnormal proliferation of white blood cells in the bone marrow, resulting in a large accumulation of abnormal leukemia cells in the blood and bone marrow. Hemorrhoids are dilated and swollen veins in the rectum or anal area. However, the relationship between CALM3 and leukemia and hemorrhoids remains unclear. The hemorrhoids dataset GSE154650 and leukemia dataset GSE26294 were downloaded from GEO databases generated by GPL20301 and GPL571.The R package limma was used to screen differentially expressed genes (DEDs). Weighted gene co-expression network analysis (WGCNA) was performed. The construction and analysis of protein-protein interaction (PPI) network, functional enrichment analysis, Gene Set Enrichment Analysis (GSEA) and comparative toxicogenomics database (CTD) analysis were performed. TargetScan was used to screen miRNAs regulating central DEGs. It was verified by western blot basic cell assay. A total of 125 DEGs were co-identified. According to the GO analysis, they are mainly enriched in small molecule catabolic processes, skin development, and chemokine receptor binding. The KEGG analysis results show that the target cells are mainly enriched in the interaction of cytokines and cytokine receptors, as well as butyric acid metabolism. The GSEA analysis results indicate enrichment in small molecule catabolic processes, skin development, and chemokine receptor binding. Six core genes (CALM3, ACE2, PPARGC1A, XCR1, CFTR, PRKCA) were identified. We found that the core gene CALM3 is highly expressed in hemorrhoid samples, low in leukemia samples, and has low expression in normal samples, which may play a regulatory role in hemorrhoids and leukemia. Immunoinfiltration results showed a higher proportion of T_cells_CD4_memory_resting and a correlation with T_cells_CD8. WB experiment verified the result. CALM3 expression is low in leukemia, and the lower the expression is, the worse the prognosis is. CALM3 is highly expressed in hemorrhoids, and the higher the expression, the worse the prognosis.

Regulation of the Hippo/YAP axis by CXCR7 in the tumorigenesis of gastric cancer.

BACKGROUND: The Hippo pathway is crucial in organ size control and tumorigenesis. Dysregulation of the Hippo/YAP axis is commonly observed in gastric cancer, while effective therapeutic targets for the Hippo/YAP axis are lacking. Identification of reliable drug targets and the underlying mechanisms that could inhibit the activity of the Hippo/YAP axis and gastric cancer progression is urgently needed. METHODS: We used several gastric cancer cell lines and xenograft models and performed immunoblotting, qPCR, and in vivo studies to investigate the function of CXCR7 in gastric cancer progression. RESULTS: In our current study, we demonstrate that the membrane receptor CXCR7 (C-X-C chemokine receptor 7) is an important modulator of the Hippo/YAP axis. The activation of CXCR7 could stimulate gastric cancer cell progression through the Hippo/YAP axis in vitro and in vivo, while pharmaceutical inhibition of CXCR7 via ACT-1004-1239 could block tumorigenesis in gastric cancer. Molecular studies revealed that the activation of CXCR7 could dephosphorylate YAP and facilitate YAP nuclear accumulation and transcriptional activation in gastric cancer. CXCR7 functions via G-protein Gαq/11 and Rho GTPase to activate YAP activity. Interestingly, ChIP assays showed that YAP could bind to the promoter region of CXCR7 and facilitate its gene transcription, which indicates that CXCR7 is both the upstream signalling and downstream target of the Hippo/YAP axis in gastric cancer. CONCLUSION: In general, we identified a novel positive feedback loop between CXCR7 and the Hippo/YAP axis, and blockade of CXCR7 could be a plausible strategy for gastric cancer.

Pegloticase co-administered with high MW polyethylene glycol effectively reduces PEG-immunogenicity and restores prolonged circulation in mouse.

The interactions between polymers and the immune system remains poorly controlled. In some instances, the immune system can produce antibodies specific to polymer constituents. Indeed, roughly half of pegloticase patients without immunomodulation develop high titers of anti-PEG antibodies (APA) to the PEG polymers on pegloticase, which then quickly clear the drug from circulation and render the gout treatment ineffective. Here, using pegloticase as a model drug, we show that addition of high molecular weight (MW) free (unconjugated) PEG to pegloticase allows us to control the immunogenicity and mitigates APA induction in mice. Compared to pegloticase mixed with saline, mice repeatedly dosed with pegloticase containing different MW or amount of free PEG possessed 4- to 12- fold lower anti-PEG IgG, and 6- to 10- fold lower anti-PEG IgM, after 3 rounds of pegloticase dosed every 2 weeks. The markedly reduced APA levels, together with competitive inhibition by free PEG, restored the prolonged circulation of pegloticase to levels observed in APA-naïve animals. In contrast, mice with pegloticase-induced APA eliminated nearly all pegloticase from the circulation within just four hours post-injection. These results support the growing literature demonstrating free PEG may effectively suppress drug-induced APA, which in turn may offer sustained therapeutic benefits without requiring broad immunomodulation. We also showed free PEG effectively blocked the PEGylated protein from binding with cells expressing PEG-specific B cell receptors. It provides a template of how we may be able to tune the interactions and immunogenicity of other polymer-modified therapeutics. STATEMENT OF SIGNIFICANCE: A major challenge with engineering materials for drug delivery is their interactions with the immune system. For instance, our body can produce high levels of anti-PEG antibodies (APA). Unfortunately, the field currently lack tools to limit immunostimulation or overcome pre-existing anti-PEG antibodies, without using broad immunosuppression. Here, we showed that simply introducing free PEG into a clinical formulation of PEG-uricase can effectively limit induction of anti-PEG antibodies, and restore their prolonged circulation upon repeated dosing. Our work offers a readily translatable method to safely and effectively restore the use PEG-drugs in patients with PEG-immunity, and provides a template to use unconjugated polymers with low immunogenicity to regulate interactions with the immune system for other polymer-modified therapeutics.

Spontaneous Redox-Reaction-Driven Growth of Ag Nanoparticles on Co(OH)2 Nanoflower Arrays for Surface-Enhanced Raman Scattering.

A simple and reliable method is developed to fabricate Ag-nanoparticle-decorated Co(OH)2 nanoflowers grafted on polyacrylonitrile (PAN) nanopillar arrays as uniform and sensitive surface-enhanced Raman scattering (SERS) substrates. First, Co(OH)2-nanosheet-assembled nanoflowers are achieved on the highly uniform PAN nanopillar arrays via electrochemical deposition. Then, Ag nanoparticles (Ag-NPs) are decorated onto the Au-nanoparticle-precoated Co(OH)2 nanoflowers based on a spontaneous redox reaction (SRR) between the silver ions and Co(OH)2 nanosheets at room temperature. Ag-NPs can be successfully in situ synthesized on the Co(OH)2 nanoflowers, and Au nanoparticles precoated on the surface of the Co(OH)2 nanosheets can ensure that the Co(OH)2 nanoflower structure does not collapse. Because of the highly uniform PAN nanopillar arrays and the high-density sub-10 nm gaps between the neighboring Ag-NPs on the surface of the Co(OH)2 nanoflowers, the hierarchical three-dimensional Ag@Co(OH)x grown on PAN nanopillar arrays can produce a reproducible and sensitive SERS effect. To verify the SERS performance of the substrate, 4-aminothiophenol (4-ATP) is used as the probe molecule, and the Ag@Co(OH)x grown on PAN nanopillar arrays is employed as the SERS substrate. As a result, 4-ATP concentrations as low as 10-10 M can still be identified, exhibiting high SERS activity. Additionally, the relative standard deviation value of the main characteristic peak of 10-5 M 4-ATP is 9.43%, indicating good uniformity of the SERS signal of the substrate. The SRR between silver ions and Co(OH)2 can provide a simple route to prepare heterostructures as SERS substrates, which has great potential for application in the field of analysis.