Zhaozhu Li

Despite significant improvements in the overall survival of Wilms tumor (WT), a subset of patients still experiences poor outcomes. Programmed cell death (PCD) pathways are pivotal in cancer progression. A deeper understanding of their roles in WT is crucial for harnessing these mechanisms to optimize risk stratification. Key tumor-associated genes were identified through limma differential analysis and WGCNA, and subsequently integrated with 12 distinct PCD patterns. TARGET-WT transcriptomic data was divided into training and validation sets to construct and validate a risk stratification model. It was subsequently integrated with clinical information to build a comprehensive prediction model. Immune infiltration and drug sensitivity analyses were performed. External validation was performed using scRNA-seq data from GSE200256. Key tumor-associated genes were enriched in multiple PCD pathways. The risk stratification model was constructed using 4 genes selected via the machine learning algorithm, stratifying the cohort into high- and low-risk groups. In the overall WT cohort, the high-risk group exhibited worse prognosis, with 1-, 3-, and 5-year AUC values of 0.820, 0.721, and 0.728, respectively. DCA demonstrated the superior predictive accuracy of the comprehensive prediction model. The high-risk group showed lower infiltration of TH17 cells and increased sensitivity to paclitaxel and sorafenib. Finally, the expression landscape of hub genes was validated in the single-cell dataset. These results highlight a critical role for PCD genes in the progression and immune regulation of WT. Targeting these genes offers a promising avenue for improving clinical management of patients identified as high-risk.

Small extracellular vesicles originating from adipose-derived mesenchymal stromal cells (ADSC-sEVs) have excellent therapeutic value in acute tendon injury

Background: Extracellular vesicles (EVs) from adipose-derived mesenchymal stem cells (ADSCs) play an important role in tendon healing and regeneration. However, the role of EVs in regulating tendon fibrosis during autophagy has not previously been reported. Purpose: To investigate the effect of EVs from ADSCs (ADSC-EVs) on the autophagy-mediated regulation of tendon fibrosis. Study Design: Controlled laboratory study. Methods: We extracted ADSCs and tenocytes from Sprague-Dawley rats, isolated EVs, and verified their uptake by tenocyts tendon fibroblasts. To assess the effects of ADSC-EVs on tendon fibroblasts, we conducted an EdU (5-ethynyl-2′-deoxyuridine) assay, scratch assay, and transwell assay to evaluate cell proliferation and migration. The activation of autophagy, a fundamental function of eukaryotic cells, by ADSC-EVs was investigated through western blotting, transmission electron microscopy, and transfection with green fluorescent protein–red fluorescent protein–LC3B. Finally, an in vivo experiment was conducted using a patellar tendon healing model. Hematoxylin and eosin staining and biomechanical testing were performed to assess tendon healing. Results: In vitro, ADSC-EVs enhanced the proliferation and migration of tendon fibroblasts after uptake. In vivo, ADSC-EVs were mixed with a gelatin methacryloyl hydrogel, applied to the injured patellar tendon, and exposed to ultraviolet irradiation for coagulation. Biomechanical testing, immunohistochemistry, and hematoxylin and eosin staining confirmed that ADSC-EVs promoted effective tendon healing. Conclusion: We demonstrated that ADSC-EVs reduced fibrosis during tendon healing by activating autophagy, thereby promoting high-quality healing. Clinical Relevance: Our findings confirm that ADSC-EVs can reduce fibrosis and inflammation during tendon healing, which may be a promising treatment approach to improve the quality of tendon healing. Therefore, ADSC-EVs have the potential to be used as cell-free biotherapeutics.

Tendon injuries are challenging to treat due to the limited blood supply and low cell density in tendon tissue. Tendon stem cells (TSCs) typically reside in a hypoxic environment (1%-5% oxygen), which may be crucial for their physiological function. In this study, we demonstrated that under hypoxic conditions, TSCs release small extracellular vesicles (sEVs) enriched with functional molecules (mainly noncoding RNAs and proteins). These sEVs significantly reduced fibrosis and inflammation in a rat model of an Achilles tendon injury while enhancing the expression of tendon-related factors, thus promoting tendon healing and improving tissue structure and function. In vitro experiments revealed that hypoxia-induced TSC-derived sEVs not only promoted TSC differentiation but also inhibited the conversion of tenocytes to myofibroblasts and reduced the expression of inflammatory markers in macrophages. These effects were mediated through the HIF-1α/miR-145-3p and miR-504 pathways. Proteomic analysis revealed hypoxia-induced changes in the protein expression profile of sEVs, with differentially expressed proteins closely associated with tendon injury repair. Our findings provide compelling evidence that hypoxia-induced sEVs from TSCs play a pivotal role in tendon repair, offering a robust theoretical basis for the development of therapeutic strategies targeting tendon injuries.

Purpose: Oxidative stress (OS) plays a key role in gastric cancer (GC). The purpose of this study was to investigate the role of the mRNA monooxygenase DBH-like 1 ( MOXD1 ) in OS and evaluate its prognostic significance in GC. Methods: An OS risk score was constructed by unsupervised clustering analysis, the log-rank test, and least absolute shrinkage and selection operator–Cox analysis of OS-related genes. The Pearson correlation between MOXD1 expression and the OS risk score was evaluated. Correlations between MOXD1 expression and clinicopathological features in the training cohort were compared. CIBERSORT, ssGSEA, and ESTIMATE were used to analyze the effects of MOXD1 on the immune microenvironment. Gene Ontology, Kyoto Encyclopedia of Genes and Genomes, and gene set enrichment analysis were used to elucidate the biological functions of the mRNAs. Immunohistochemistry for MOXD1 was performed on patient tissue microarray (TMA) samples. Cox regression, log-rank tests, and chi-square analyses were used to investigate the clinicopathological features of the TMAs and associated MOXD1 expression levels. A stable knockdown cell line was constructed in HGC-27 GC cells and investigated using cell counting kit-8 and Transwell assays. Results: The OS risk score was an independent prognostic factor for GC in the training cohort and was successfully combined with age and pTNM stage to construct a nomogram. MOXD1 expression was positively correlated with the OS risk score and was highly expressed in patients with GC. MOXD1 expression and the metastatic lymph node ratio in TMAs were found to be independent prognostic risk factors for GC. MOXD1 knockdown inhibited the proliferation and invasion of HGC-27 cells. Conclusion: The mRNA MOXD1 is a biomarker for both OS and GC. MOXD1 expression can be used to evaluate GC prognosis and guide treatment.

Background Pediatric inguinal hernia is a common surgical condition among children. In this study, we present the application of a self-designed laparoscopic auxiliary needle device (China Patent Number: ZL 201320479515.5) and evaluate its effectiveness in laparoscopic percutaneous extraperitoneal closure in pediatric patients. Methods The laparoscopic auxiliary needle device, with a diameter of 1 mm, consists of a cylindrical needle body and a specifically designed front end. Between January 2013 and January 2021, a total of 2075 children with inguinal hernia (1864 males and 211 females, mean age 3.71 ± 2.95 years old) were included in this study. All patients underwent laparoscopic inguinal hernia repair using the laparoscopic auxiliary needle device. Various data were collected including occult inguinal hernia incidence, complications such as recurrence, and follow-up findings. Results The entire patient cohort underwent laparoscopic inguinal hernia repair using the laparoscopic auxiliary needle. Pneumoperitoneum lasted 5.1 ± 1.2 min in unilateral hernia cases and 11.0 ± 1.3 min in bilateral hernia cases. Occult inguinal hernia were found in 981 (47.3%) patients, out of which initially difficult diagnoses were encountered in 516 cases (24.9%). Postoperative follow-up ranged from 3 to 96 months, with two instances (0.1%) of inguinal hernia recurrence reported. Importantly, no patients reported any visible scars or sensations of subcutaneous foreign bodies. Conclusion Based on our extensive patient cohort, we assert that our self-designed laparoscopic auxiliary needle device represents a valuable tool for performing laparoscopic inguinal hernia repairs while minimizing abdominal scarring. Additionally, this device offers the additional advantage of detecting contralateral occult inguinal hernia. Graphical abstract

Background: Wilms’ tumor (WT) is the most common renal tumor in childhood. Pyroptosis, a type of inflammation-characterized and immune-related programmed cell death, has been extensively studied in multiple tumors. In the current study, we aim to construct a pyroptosis-related gene signature for predicting the prognosis of Wilms’ tumor. Methods: We acquired RNA-seq data from TARGET kidney tumor projects for constructing a gene signature, and snRNA-seq data from GEO database for validating signature-constructing genes. Pyroptosis-related genes (PRGs) were collected from three online databases. We constructed the gene signature by Lasso Cox regression and then established a nomogram. Underlying mechanisms by which gene signature is related to overall survival states of patients were explored by immune cell infiltration analysis, differential expression analysis, and functional enrichment analysis. Results: A pyroptosis-related gene signature was constructed with 14 PRGs, which has a moderate to high predicting capacity with 1-, 3-, and 5-year area under the curve (AUC) values of 0.78, 0.80, and 0.83, respectively. A prognosis-predicting nomogram was established by gender, stage, and risk score. Tumor-infiltrating immune cells were quantified by seven algorithms, and the expression of CD8( +) T cells, B cells, Th2 cells, dendritic cells, and type 2 macrophages are positively or negatively correlated with risk score. Two single nuclear RNA-seq samples of different histology were harnessed for validation. The distribution of signature genes was identified in various cell types. Conclusions: We have established a pyroptosis-related 14-gene signature in WT. Moreover, the inherent roles of immune cells (CD8( +) T cells, B cells, Th2 cells, dendritic cells, and type 2 macrophages), functions of differentially expressed genes (tissue/organ development and intercellular communication), and status of signaling pathways (proteoglycans in cancer, signaling pathways regulating pluripotent of stem cells, and Wnt signaling pathway) have been elucidated, which might be employed as therapeutic targets in the future.

Background:Fibrosis and inflammation due to ureteropelvic junction obstruction substantially contributes to poor renal function. Urine-derived stem-cell-derived exosomes (USC-Exos) have therapeutic effects through paracrine.Methods:In vitro, the effects of USC-Exos on the biological functions of HK-2 and human umbilical vein endothelial cells were tested. Cell inflammation and fibrosis were induced by transforming growth factor-β1 and interleukin-1β, and their anti-inflammatory and antifibrotic effects were observed after exogenous addition of USC-Exos. Through high-throughput sequencing of microRNA in USC-Exos, the pathways and key microRNAs were selected. Then, the antifibrotic and anti-inflammatory effects of exosomal miR-122-5p and target genes were verified. The role of the miR-122-5p/SOX2 axis in anti-inflammatory and antifibrotic effects was verified. In vivo, a rabbit model of partial unilateral ureteral obstruction (PUUO) was established. Magnetic resonance imaging recorded the volume of the renal pelvis after modeling, and renal tissue was pathologically analyzed.Results:We examined the role of USC-Exos and their miR-122-5p content in obstructive kidney injury. These Exos exhibit antifibrotic and anti-inflammatory activities. SOX2 is the hub gene in PUUO and negatively related to renal function. We confirmed the binding relationship between miR-122-5p and SOX2. The anti-inflammatory and antifibrotic effects of miR-122-5p were inhibited, indicating that miR-122-5p has anti-inflammatory and antifibrotic effects by inhibiting SOX2 expression. In vivo, the PUUO group showed typical obstructive kidney injury after modeling. After USC-Exo treatment, the shape of the renal pelvis shown a remarkable improvement, and inflammation and fibrosis decreased.Conclusions:We confirmed that miR-122-5p from USC-Exos targeting SOX2 is a new molecular target for postoperative recovery treatment of obstructive kidney injury.