Gastric injury repair poses a significant clinical challenge due to the ongoing exposure to gastric acid, complex mechanical environments, and intense inflammatory responses. To address these issues, we developed a bioactive asymmetric Janus hydrogel (PSE/GDMA-BPN@Ori) that features strong wet adhesion, immunomodulatory properties, and regenerative potential for repairing gastric injuries. This hydrogel consists of a bilayer: an anti-adhesive layer made from electrospun polycaprolactone/decellularized small intestinal submucosa (PCL/dSIS-ECM) and a dopamine-modified gelatin methacrylate (GDMA) adhesive layer, which is functionalized with black phosphorus nanosheets (BPN) and loaded with the natural flavonoid orientin (Ori). This configuration allows for one-sided wet adhesion, mechanical robustness, and controlled drug release. The hydrogel demonstrates excellent biocompatibility, antioxidant capacity, hemostatic performance, and in vivo biodegradation. In vitro studies show that it effectively scavenges reactive oxygen species and promotes macrophage polarization toward the M2 reparative phenotype. In a rat model of gastric perforation, the hydrogel significantly accelerated mucosal regeneration, enhanced angiogenesis, and reduced postoperative inflammation and adhesion formation. Overall, this multifunctional Janus adhesive hydrogel integrates wet adhesion, immune regulation, and bioactive repair, facilitating rapid and stable gastric tissue healing. It presents a promising biomaterial alternative for treating gastric perforations and anastomotic leakage.

The aim of this study was to investigate whether eprenetapopt (APR-246, PRIMA-1MET) could enhance anti-tumor effects of venetoclax (VEN) on cell proliferation and apoptosis in vitro cell lines, and to further explore the potential mechanisms underlying the enhanced effects. The cellular proliferative capacity was assessed using the CCK-8 assay, whereas apoptosis levels were evaluated through Western blot analysis and flow cytometry. Key pathways and genes were identified using the following bioinformatics tools: the Cancer Dependency Map (DepMap) and the Cancer Genome Atlas Program (TCGA) database, gene set enrichment analysis (GSEA), and the Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. Both APR-246 and VEN monotherapy demonstrated significant anti-proliferative and pro-apoptotic effects in TP53 R248Q-mutated AML cell lines. Notably, APR-246 enhanced therapeutic efficacy of VEN in TP53-mutated AML cells. From a mechanistic perspective, APR-246 may exert its anti-leukemic effects through dual pathways: (1) Associated with downregulation of PPP1CA expression, which could potentially involve the Hippo signaling pathway, coinciding with observed proliferation inhibition and apoptosis induction; (2) Attenuation of VEN-induced MCL-1 (myeloid cell leukemia-1) overexpression, potentially contributing to the augmented anti-tumor activity with VEN in TP53-mutated AML models. In TP53 R248Q-mutated AML cell lines, APR-246 augmented anti-tumor effects of VEN via proliferation inhibition and apoptosis induction. These observed enhanced effects may relate to multiple factors including: PPP1CA inhibition, Hippo pathway affecting, and MCL-1 expression suppression, warranting further investigation.

Astragaloside IV (AS-IV), a key active component derived from the traditional Chinese medicinal plant Astragali, has been reported to exhibit various biological activities, including antioxidative, anti-inflammatory, immunoregulatory, and antineoplastic properties. This study aimed to elucidate the role of AS-IV in inhibiting gastric cancer (GC) growth, focusing on its impact on cell ferroptosis and the underlying molecular mechanisms. Proliferation and migration of GC cells upon AS-IV treatment were examined using CCK-8, colony formation, and Transwell assays. Ferroptosis induction was analyzed via ELISA, flow cytometry, and transmission electron microscopy. Ferroptosis suppressor protein 1 (FSP1) mRNA stability was assessed by the ActD assay, while RNA immunoprecipitation (RIP) was employed to confirm the interaction between FSP1 mRNA, Fat mass and obesity-associated protein (FTO) demethylase, and YTH N6-methyladenosine RNA-binding protein F2 (YTHDF2). The dual-luciferase reporter assay was used to explore FTO binding to N6-methyladenosine (m6A)-modified sites on FSP1 mRNA. Furthermore, AS-IV’s anti-tumor effects (20 mg/kg) were validated in vivo using gastric cancer xenograft and lung metastasis mouse models. AS-IV significantly suppressed the proliferation and migration of GC cells by inducing ferroptosis. Mechanistically, AS-IV down-regulated FTO, thus impairing its interaction with FSP1 mRNA and leading to increased m6A modification on FSP1 mRNA. This modification facilitated m6A recognition protein YTHDF2-mediated recognition and subsequent degradation of FSP1 mRNA. The reduction of FSP1 triggered ferroptosis, while the overexpression of FSP1 or inhibition of ferroptosis by ferrostatin-1 partially reversed AS-IV’s effects on cell viability and migration. In vivo, AS-IV effectively inhibited tumor growth and metastasis. This study highlights potent anti-GC effects of AS-IV, mediated by the suppression of FSP1 via the FTO/YTHDF2/m6A axis. The treatment of AS-IV inhibits the expression of demethylase FTO in GC cells, subsequently disrupting the binding between FTO protein and FSP1 mRNA. This disruption leads to a increase in the level of m6A modification on FSP1 mRNA, thereby enhancing the recognition and binding of m6A-recognizing protein YTHDF2, promoting the decay of FSP1 mRNA. Ultimately, the downregulation of FSP1 enhances ferroptosis in GC cells. Collectively. AS-IV holds great promise as a novel therapeutic strategy for gastric cancer treatment. This image was created using the Biorender website (https://www.biorender.com/).

CAR-T cell therapy, as a representative technology in cancer immunotherapy, has demonstrated notable success in the treatment of hematologic malignancies; however, a significant proportion of patients fail to achieve sustained remission. Through the analysis of bone marrow sequencing data prior to CD19 CAR-T cell therapy, we identified cellular adhesion as a pivotal factor influencing clinical outcomes. We developed a model to predict B-ALL treatment efficacy based on the core genes associated with cellular adhesion, which was validated in our clinical cohort. Both in vitro and in vivo experiments revealed that the inhibition or knockout of integrin subunit beta 2 (ITGB2, also known as CD18) in malignant B cells markedly diminished the cytotoxic efficacy of both CD19 and CD20 CAR-T cells against B-lineage tumor cells, with alterations in ITGB2-mediated cytotoxicity linked to the formation of immunological synapses within the tumor microenvironment. Notably, the upregulation of ITGB2 in Nalm6 cells via LPS or Venetoclax significantly augmented the cytotoxic activity of CAR-T cells against Nalm6 cells. Our findings provide a novel predictive model for clinical CD19 CAR-T cell therapy and elucidate a role of the ITGB2 pathway in CAR-T cell-mediated eradication of B-cell malignancies.

Although chimeric antigen receptor (CAR)-redirected T lymphocytes have achieved unprecedented clinical responses in hematologic neoplasms, their role in solid tumors still needs to be improved. To overcome current limitations, we rationally designed a next-generation CAR-T cell construct incorporating autonomous costimulatory signaling pathways that operate independently of tumor antigen engagement, thus faithfully emulating physiological T cell activation paradigms. We screened seven costimulatory receptors from the TNF receptor superfamily and identified that CD30 is the most effective enhancer of CAR-T cell function. Subsequent structural analysis revealed that the intracellular domain (ICD) of CD30 is primarily responsible for its costimulatory activity. Our data showed that these CAR-T cells co-expressing CD30 ICD have stronger proliferation ability and cytokine secretion function. The results of in vitro experiments showed that CD30 signaling improved the cytotoxicity of CAR-T cells and reduced the expression of exhaustion-related markers. In mouse models of orthotopic renal cancer, lung metastasis, and hematologic malignancies, these CAR-T cells showed better expansion capability and superior antitumor activity. Sequencing results suggest that CD30 signaling may enhance the function of CAR-T cells by increasing the activation of the nuclear factor κB (NF-κB) pathway. Further mechanistic experiments confirmed that the NF-κB pathway is significantly activated in CAIX.CD30(ICD) CAR-T cells compared to CAIX CAR-T cells. Reversal experiments demonstrated that NF-κB pathway inhibitors can reverse the effector function of CAIX.CD30(ICD) CAR-T cells, while CAIX-IKKβ CAR-T cells with self-activated NF-κB pathway also exhibit significant functional advantages. By integrating autonomous costimulatory signaling, we demonstrated improved CAR-T cell persistence, proliferation, and antitumor activity across multiple preclinical models, highlighting the therapeutic potential of this approach for both hematologic malignancies and solid tumors.

Clear cell renal cell carcinoma (ccRCC), the most common type of renal cell carcinoma (RCC), is not sensitive to traditional radiotherapy and chemotherapy. The polyphenolic compound Gallic acid (GA) can be naturally found in a variety of fruits, vegetables and plants. Autophagy, an intracellular catabolic process, regulates the lysosomal degradation of organelles and portions in cytoplasm. It was reported that autophagy and GA could affect the development of several cancers. Therefore, the aim of the present study was to evaluate the effects of GA on ccRCC development and clarify the role of autophagy in this process. In the present study, the effects of GA on the proliferation, migration and invasion of ccRCC cells were investigated in vitro by Cell Counting Kit‑8, colony formation, flow cytometry, wound healing and Transwell migration assays, respectively. Additionally, the effects of GA on ccRCC growth and metastasis were evaluated using hematoxylin‑eosin and immunohistochemical staining in vivo. Moreover, it was sought to explore the underlying molecular mechanisms using transmission electron microscopy, western blotting and reverse transcription‑quantitative PCR analyses. In the present study, it was revealed that GA had a more potent viability inhibitory effect on ccRCC cells (786‑O and ACHN) than the effect on normal renal tubular epithelial cell (HK‑2), which demonstrated that GA selectively inhibits the viability of cancer cells. Furthermore, it was identified that GA dose‑dependently inhibited the proliferation, migration and invasion of ccRCC cells in vitro and in vivo. It was demonstrated that GA promoted the release of autophagy markers, which played a role in regulating the PI3K/Akt/Atg16L1 signaling pathway. All the aforementioned data provided evidence for the great potential of GA in the treatment of ccRCC.

This study aimed to identify novel differentially expressed genes in breast cancer and to explore the clinical value and the anti-tumor or oncogenic effects of the identified genes using bioinformatics analysis and in vitro experiments. The differentially expressed genes in breast cancer patients were identified using Gene Expression Omnibus (GEO) database with the cut-off criteria p < 0.05 and |logFC| > 1. The expression levels of palmdelphin (PALMD) and dermatopontin (DPT) in normal tissues and breast cancer tissues were evaluated based on GEPIA and UALCAN databases. PALMD and DPT expression levels in clinical subgroups of patients with breast cancer were analyzed to assess the association of PALMD and DPT expression with clinical characteristics. The prognostic and diagnostic values of PALMD and DPT in breast cancer were evaluated from Kaplan-Meier (K-M) survival curves and receiver operating characteristic (ROC) curves. Pearson's correlation coefficient was performed using LinkedOmics. KEGG pathway enrichment analysis was performed using DAVID. The protein levels were evaluated using western blot analysis. Cell proliferation was assessed using MTT and EdU assays. Two important genes, PALMD and DPT, were identified in breast cancer. The expression levels of PALMD and DPT were significantly lower in breast cancer tissues. The expression levels of PALMD were closely related to age, histological type, and T stage of breast cancer patients. The expression levels of DPT were closely related to age, histological type, T stage, N stage, estrogen receptor status, and progesterone receptor status of breast cancer patients. The K-M survival curves showed that PALMD or DPT was not an independent prognostic factor for breast cancer. The ROC curves showed that both PALMD and DPT had good diagnostic potential for breast cancer. KEGG pathway enrichment results showed that PI3K/Akt pathway was an important overlapping signaling for PALMD and DPT. Further studies proved that overexpression of PALMD and DPT inhibited proliferation in MCF-7 and MDA-MB-231 cells by suppressing the PI3K/Akt pathway. PALMD and DPT knockdown promoted proliferation in MCF-7 and MDA-MB-231 cells by activating the PI3K/Akt pathway. These results collectively suggested that PALMD and DPT might serve as potential diagnostic biomarkers and therapeutic targets for breast cancer.

Clear cell renal cell carcinoma (ccRCC) has a high metastatic rate, and its incidence and mortality are still rising. The aim of this study was to identify the key tumor-infiltrating immune cells (TIICs) affecting the distant metastasis and prognosis of patients with ccRCC and to construct a relevant prognostic panel to predict immunotherapy response. Based on ccRCC bulk RNA sequencing data, resting mast cells (RMCs) were screened and verified using the CIBERSORT algorithm, survival analysis, and expression analysis. Distant metastasis-associated genes were identified using single-cell RNA sequencing data. Subsequently, a three-gene (CFB, PPP1R18, and TOM1L1) panel with superior distant metastatic and prognostic performance was established and validated, which stratified patients into high- and low-risk groups. The high-risk group exhibited lower infiltration of RMCs, higher tumor mutation burden (TMB), and worse prognosis. Therapeutically, the high-risk group was more sensitive to anti-PD-1 and anti-CTLA-4 immunotherapy, whereas the low-risk group displayed a better response to anti-PD-L1 immunotherapy. Furthermore, two immune clusters revealing distinct immune, clinical, and prognosis heterogeneity were distinguished. Immunohistochemistry of ccRCC samples verified the expression patterns of the three key genes. Collectively, the prognostic panel based on RMCs is able to predict distant metastasis and immunotherapy response in patients with ccRCC, providing new insight for the treatment of advanced ccRCC.

Previous studies have identified the dysregulation of various circRNAs in many types of human cancers including thyroid cancer (TC). Circular RNA ZFR (circZFR) serves as an oncogenic circRNA in TC. However, the detailed molecular mechanism of circZFR in TC progression remains to be further explored. CircZFR and miR-16 expressions in TC cells were analyzed through qRT-PCR. Cell viability, invasion, and apoptosis were detected using CCK-8, transwell invasion assay, and flow cytometry analysis, respectively. The relationship between circZFR and miR-16 was explored using luciferase reporter assay, RNA pull-down assay, and qRT-PCR. The relationship between miR-16 and mitogen-activated protein kinase 1 (MAPK1) was explored using luciferase reporter assay and western blot analysis. Results showed that circZFR was upregulated and miR-16 was downregulated in TC cells. CircZFR knockdown inhibited the viability and invasion and induced apoptosis in TC cells. CircZFR inhibited miR-16 expression by sponging miR-16 and miR-16 repressed MAPK1 expression by targeting MAPK1. Moreover, circZFR positively regulated MAPK1 expression in TC cells by serving as a ceRNA of miR-16. Mechanistically, circZFR knockdown-induced inhibition of cell viability and invasion and promotion of apoptosis were overturned after miR-16 downregulation and promotion of MAPK1. Collectively, circZFR knockdown retarded TC progression by sponging miR-16 and modulating MAPK1 expression.

Clear cell renal cell carcinoma (ccRCC) is a common malignancy with high distant metastasis rate. Long non-coding RNAs (LncRNAs) are reported to be upregulated or downregulated in multiple cancers and play a crucial role in the metastasis of tumors or prognosis. Therefore, the purpose of our study is to construct a prognostic signature for ccRCC based on distant metastasis-related lncRNAs and explore the involved potential competitive endogenous RNA (ceRNA) network. The differentially expressed genes (DEGs) screened from the database of the cancer genome atlas (TCGA) were used to construct a co-expression network and identify the distant metastasis-related module by weighted gene co-expression network analysis (WGCNA). Key genes with metastatic and prognostic significance were identified through rigorous screening, including survival analysis, correlation analysis, and expression analyses in stage, grade, and distant metastasis, and were verified in the data set of gene expression omnibus (GEO) and the database from gene expression profiling interactive analysis (GEPIA). The potential upstream miRNAs and lncRNAs were predicted via five online databases and LncBase. Here, we constructed a ceRNA network of key genes that are significantly associated with the distant metastasis and prognosis of patients with ccRCC. The distant metastasis-related lncRNAs were used to construct a risk score model through the univariate, least absolute shrinkage selection operator (LASSO), and multivariate Cox regression analyses, and the patients were divided into high- and low-risk groups according to the median of the risk score. The Kaplan–Meier survival analysis demonstrated that mortality was significantly higher in the high-risk group than in the low-risk group. Considering the other clinical phenotype, the Cox regression analyses indicated that the lncRNAs model could function as an independent prognostic factor. Quantitative real-time (qRT)-PCR in the tissues and cells of ccRCC verified the high-expression level of three lncRNAs. Gene set enrichment analysis (GSEA) revealed that the lncRNA prognostic signature was mainly enriched in autophagy- and immune-related pathways, indicating that the autophagy and immune functions may play an important role in the distant metastasis of ccRCC. In summary, the constructed distant metastasis-related lncRNA signature could independently predict prognosis in patients with ccRCC, and the related ceRNA network provided a new sight on the potential mechanism of distant metastasis and a promising therapeutic target for ccRCC.

Circular RNAs (circRNAs) have been documented to be aberrantly expressed in many types of malignancies and involved in cancer progression. However, their role in thyroid cancer (TC) remains largely unknown. Our study aimed to explore the role and mechanism of circUBAP2 in TC. The differentially expressed circRNAs in TC tissues were identified using GSE18105 from gene expression omnibus (GEO) database. CircUBAP2 and miR-370-3p expression was analyzed using qRT-PCR. The stability of circUBAP2 was confirmed by actinomycin D and RNase R. The subcellular localization of circUBAP2 was detected using cell fractionation assay. Cell proliferation, apoptosis, and invasion were evaluated using MTT, flow cytometry analysis, and Transwell invasion assay, respectively. The interaction between circUBAP2 and miR-370-3p was predicted using bioinformatics analysis and validated by luciferase reporter assay, RNA pull-down assay, and RNA immunoprecipitation. CircUBAP2 was upregulated and miR-370-3p was downregulated in TC tissues and cells. CircUBAP2 was highly stable, resistant to RNase R digestion, and predominantly localized in the cytoplasm. CircUBAP2 knockdown inhibited cell proliferation and invasion and triggered apoptosis in TC cells. Bioinformatics analysis showed that circUBAP2 contained putative binding sites of miR-370-3p. CircUBAP2 acted as a sponge to inhibit miR-370-3p expression. Mechanistically, miR-370-3p inhibition abolished the effects of circUBAP2 on proliferation, apoptosis, and invasion in TC cells. Taken together, CircUBAP2 knockdown impeded the proliferation and invasion and induced apoptosis in TC cells via sponging miR-370-3p.

Background: Recently, some lncRNAs in studies were also regarded as the potential prognostic biomarkers for renal cell carcinoma. The aim of this meta-analysis is to evaluate the prognostic significance of lncRNAs for RCC and explore their correlation with lymph node metastasis (LNM) and distant metastasis (DM). Methods: A series of electronic databases were chosen to retrieve the eligible articles. A total of 35 studies involving 3535 patients with RCC were included finally. The statistical analysis was performed using STATA 16.0 and review manager 5.3 software. The relative risk (RR) and hazard ratio (HR) with 95% confidence interval (CI) were calculated to assess the significance. The stratified subgroup analysis was conducted by the up-regulated group and down-regulated group. Additionally, the sensitivity analysis was performed by the sequential omission of individual studies, and the publication bias was detected using Begg’s test. Results: The results indicated lncRNAs expression levels were correlated with LNM for up-regulated lncRNAs (RR=2.06, 95% CI: 1.53-2.78, p<0.00001) and down-regulated lncRNAs (RR=0.49, 95% CI: 0.31-0.76, p=0.002). The expression levels of lncRNAs were closely related to DM (RR=1.67, 95% CI: 1.21-2.29, p=0.002). Moreover, the expression levels of these lncRNAs were also associated with the overall survival (HR=1.71, 95% CI: 1.28-2.28, P=0.0003). Conclusions: This meta-analysis suggested that lncRNAs could be regarded as biomarkers for clinical lymph node metastasis or distant metastasis and also may serve as the potential predictive factors of prognosis for RCC.

Accumulating evidence shows that exosomal circRNAs reflect the physiological status of donor cells, and various cell reactions are induced after exosomal circRNAs are captured by recipient cells. In this study, qRT-PCR was performed to detect circ-0004277 expression in hepatocellular carcinoma (HCC) cell lines, tissues, and plasma exosomes. The effects of circ-0004277 on the proliferation and migration of HCC cells were assessed by cell counting, 5-ethynyl-2′-deoxyuridine assays, Transwell migration assays, and tumor formation in nude mice. We found that circ-0004277 was significantly upregulated in HCC cells, tissues, and plasma exosomes compared to that in normal controls. Overexpression of circ-0004277 enhanced the proliferation, migration, and epithelial-mesenchymal transition (EMT) of HCC cells in vivo and in vitro . Furthermore, exosomes from HCC cells enhanced circ-0004277 expression in surrounding normal cells and stimulated EMT progression. ZO-1, a tight junction adapter protein, was downregulated in HCC tissues. In conclusion, our findings suggest that circ-0004277 promotes the malignant phenotype of HCC cells via inhibition of ZO-1 and promotion of EMT progression. In addition, exosomal circ-0004277 from HCC cells stimulates EMT of peripheral cells through cellular communication to further promote the invasion of HCC into normal surrounding tissues.

The mechanism by which miR‐605‐3p regulates hepatocellular carcinoma (HCC) metastasis has not been clarified. In this study, we found that miR‐605‐3p was down‐regulated in HCC and that low miR‐605‐3p expression was associated with tumour thrombus and tumour satellites. HCC patients with low miR‐605‐3p expression showed shorter overall survival and disease‐free survival after surgery. Overexpression of miR‐605‐3p inhibited epithelial‐mesenchymal transition and metastasis of HCC through NF‐κB signalling by directly inhibiting expression of TRAF6 , while silencing of miR‐605‐3p had the opposite effect. We also found that SNHG16 directly bound to miR‐605‐3p as a competing endogenous RNA. Mechanistically, high expression of SNHG16 promoted binding to miR‐605‐3p and inhibited its activity, which led to up‐regulation of TRAF6 and sustained activation of the NF‐κB pathway, which in turn promoted epithelial‐mesenchymal transition and metastasis of HCC. TRAF6 increased SNHG16 promoter activity by activating NF‐κB, thereby promoting the transcriptional expression of SNHG16 and forming a positive feedback loop that aggravated HCC malignancy. Our findings reveal a mechanism for the sustained activation of the SNHG16 / miR‐605‐3p / TRAF6 /NF‐κB feedback loop in HCC and provide a potential target for a new HCC treatment strategy.

Immune status affects the initiation and progression of clear cell renal cell carcinoma (ccRCC), the most common subtype of renal cell carcinoma. In this study, we identified an immune-related, five-gene signature that improves survival prediction in ccRCC. Patients were classified as high- and low-risk based on the signature risk score. Survival analysis showed differential prognosis, while principal component analysis revealed distinctly different immune phenotypes between the two risk groups. High-risk patients tended to have advanced stage, higher grade disease, and poorer prognoses. Functional enrichment analysis showed that the signature genes were mainly involved in the cytokine-cytokine receptor interaction pathway. Moreover, we found that tumors from high-risk patients had higher relative abundance of T follicular helper cells, regulatory T cells, and M0 macrophages, and higher expression of PD-1, CTLA-4, LAG3, and CD47 than low-risk patients. This suggests our gene signature may not only serve as an indicator of tumor immune status, but may be a promising tool to select high-risk patients who may benefit from immune checkpoint inhibitor therapy. Multivariate Cox regression analysis showed that the signature remained an independent prognostic factor after adjusting for clinicopathological variables, while prognostic accuracy was further improved after integrating clinical parameters into the analysis.

Hirschsprung disease (HSCR) is a birth defect with an approximate incidence of 1/5,000 live births, and up to one-third of HSCR patients develop Hirschsprung-associated enterocolitis (HAEC), the leading cause of HSCR-related death. Very little is known about the pathogenesis, prevention, and early diagnosis of HAEC. Here, we used a prospective study to investigate the enteric microbiome composition at the time of surgery as a predictor for developing postoperative HAEC. We identified a microbiome signature containing 21 operational taxonomic units (OTUs) that can potentially predict postoperative HAEC with ~85% accuracy. Furthermore, we identified exclusive breastfeeding as a novel protective factor for total HAEC (i.e., preoperative and postoperative HAEC combined). In addition, we discovered that breastfeeding was associated with a lowered risk for HAEC potentially mediated by modulating the gut microbiome composition characterized by a lower abundance of Gram-negative bacteria and lower LPS concentrations. In conclusion, modulating the gut microbiome by encouraging breastfeeding might prevent HAEC progression in HSCR patients.

Background: Hirschsprung-associated enterocolitis (HAEC) is the leading cause of mortality in Hirschsprung's disease (HSCR) patients. We aimed to examine the association between exclusive breastfeeding and risk of HAEC as well as the underlying mechanisms. Methods: In a hospital-based cohort study of 253 HSCR patients, we examined the association between exclusive breastfeeding and risk of HAEC by log-binomial regression. Using enteric tissue samples collected at surgery, we further conducted a prospective nested case-control study of 25 postoperative HAEC cases and 50 HSCR controls. We characterized the enteric microbiome by 16S rRNA gene sequencing and measured lipopolysaccharide by an enzyme-linked immunosorbent assay. Relative abundance of Gram-negative bacteria and LPS concentrations were compared between groups: (1) postoperative HAEC cases and controls; (2) HSCR patients with and without exclusive breastfeeding, using Wilcoxon rank-sum test. Stepwise random forest classification was used to identify an enteric microbiome signature to predict postoperative HAEC occurrence. Findings: Exclusive breastfeeding was associated with lower risk of overall and postoperative HAEC, with an adjusted RR of 0.61 (95% CI, 0.44-0.85; P=0.003) and 0.51 (95% CI, 0.24-1.08; P=0.08), respectively. Exclusive breastfeeding contributed to an enteric microbiome characterized by lower abundance of Gram-negative bacteria (particularly Enterobacteriaceae) and lower lipopolysaccharide concentrations, which was associated with lower occurrence of postoperative HAEC. In addition, we identified a microbiome signature that predicted postoperative HAEC occurrence with ~85% accuracy. Interpretation: Exclusive breastfeeding reduces risk of HAEC through modulation of the enteric microbiome. Funding Statement: This work was supported by the National Natural Science Foundation of China (NSFC 81570467). Declaration of Interests: Authors declare that they have no competing interests.Ethics Approval Statement: The study was approved by the Medicine Ethics Committee at Children’s Hospital of Nanjing Medical University and the Human Subjects Committee at the Lawrence Berkeley National Laboratory, and all participants provided informed consent.

Objective Renal cell carcinoma (RCC) is the most common malignancy of the urinary system, and it is a serious threat to human health. HOXA transcript at the distal tip (HOTTIP), located at the 5' end of the HOXA locus, is a long non-coding RNA that has been newly discovered in recent years. It has been reported to promote the development of several types of tumors. Moreover, accumulating evidence has indicated that autophagy plays an important role in tumor cell survival or death. However, whether HOTTIP affects RCC development by regulating autophagy remains unknown. Methods In this study, we first measured HOTTIP expression in 42 paired RCC and adjacent non-tumor tissues, as well as in 4 RCC cell lines and 1 normal renal tubular epithelial cell line. Then, we selected 2 RCC cell lines to silence HOTTIP expression and 1 RCC cell line to overexpress HOTTIP, and we measured their proliferation, migration and invasion, as well as autophagy, after pretreatment with an autophagy inhibitor or inducer. In addition, we assessed the growth, metastasis and autophagy of tumors in nude mice and explored the mechanism involved. Results The results showed that HOTTIP expression was significantly upregulated in the RCC tissues and cell lines, and it was closely associated with TNM stage, histological grade, lymph node metastasis and patient prognosis. The in vitro and in vivo assays indicated that HOTTIP silencing inhibited RCC cell proliferation, migration and invasion and induced autophagy, and 3-MA (an autophagy inhibitor) reversed these effects. In contrast, HOTTIP overexpression and rapamycin (an autophagy inducer) yielded the opposite results. Further research revealed that HOTTIP modification could affect RCC cell autophagy via the PI3K/Akt/Atg13 signaling pathway. Conclusions Our study will help in finding a potential marker for RCC diagnosis and supply a target molecule for RCC treatment.

Research over the past decade suggested critical roles for circular RNAs in the natural growth and disease progression. However, it remains poorly defined whether the circular RNAs participate in Hirschsprung disease (HSCR). Here, we reported that the cir-ZNF609 was down-regulated in HSCR compared with normal bowel tissues. Furthermore, suppression of cir-ZNF609 inhibited the proliferation and migration of cells. We screened out several putative cir-ZNF609 ceRNAs of which the AKT3 transcript was selected. Finally, RNA immunoprecipitation and luciferase reporter assays demonstrated that cir-ZNF609 may act as a sponge for miR-150-5p to modulate the expression of AKT3. In conclusion, these findings illustrated that cir-ZNF609 took part in the onset of HSCR through the crosstalk with AKT3 by competing for shared miR-150-5p.

Background It has been proposed that lncRNAs, widely transcribed from genomes, play pivotal regulatory roles in a variety of biological processes, but their function in regulating spermatogenesis in human males is rarely reported. Methods QRT-PCR was adopted to detect HOTTIP expression level in testicular tissues from hypospermatogenesis (Hypo) patients or controls. The proliferation levels of NT2 and 293T were measured via CCK-8 and EdU detection. Meanwhile, luciferase reporter gene assay and bioinformatics analysis were carried out to identify a target of HOTTIP. Additionally, the underlying mechanism of HOTTIP's function was investigated using western blotting and RIP analysis. Results The research results manifested that the expression of HOTTIP in testicular tissues from Hypo patients was prominently reduced in comparison with that in control testicular tissues. Interestingly, it was noted that HOTTIP exhibited a high expression in testicular embryonal carcinoma cell line NT2 compared with that in normal control cell line 293T. It was denoted in cell function evaluation that cell proliferation was impeded by downregulated HOTTIP but evidently stimulated by overexpressed HOTTIP. Moreover, HOTTIP was capable of positively modulating HOXA13 expression via the competitive binding to miR-128-3p. Conclusion Therefore, HOTTIP acting as ceRNAs to promote testicular embryonal carcinoma cell proliferation.