Comprehensive analysis of the relationship between ubiquitin-specific protease 21 (USP21) and prognosis, tumor microenvironment infiltration, and therapy response in colorectal cancer.

BACKGROUND: Ubiquitin-specific proteases family is crucial to host immunity against pathogens. However, the correlations between USP21 and immunosurveillance and immunotherapy for colorectal cancer (CRC) have not been reported. METHODS: The differential expression of USP21 between CRC tissues and normal tissues was analyzed using multiple public databases. Validation was carried out in clinical samples through qRT-PCR and IHC. The correlation between USP21 and the prognosis, as well as clinical pathological characteristics of CRC patients, was investigated. Moreover, cell models were established to assess the influence of USP21 on CRC growth and progression, employing CCK-8 assays, colony formation assays, and wound-healing assays. Subsequently, gene set variation analysis (GSVA) was used to explore the potential biological functions of USP21 in CRC. The study also examined the impact of USP21 on cytokine levels and immune cell infiltration in the tumor microenvironment (TME). Finally, the effect of USP21 on the response to immunotherapy and chemotherapy in CRC was analyzed. RESULTS: The expression of USP21 was significantly upregulated in CRC. High USP21 is correlated with poor prognosis in CRC patients and facilitates the proliferation and migration capacities of CRC cells. GSVA indicated an association between low USP21 and immune activation. Moreover, low USP21 was linked to an immune-activated TME, characterized by high immune cell infiltration. Importantly, CRC with low USP21 exhibited higher tumor mutational burden, high PD-L1 expression, and better responsiveness to immunotherapy and chemotherapeutic drugs. CONCLUSION: This study revealed the role of USP21 in TME, response to therapy, and clinical prognosis in CRC, which provided novel insights for the therapeutic application in CRC.

Comprehensive analyses of solute carrier family members identify SLC12A2 as a novel therapy target for colorectal cancer.

As the largest transporter family impacting on tumor genesis and development, the prognostic value of solute carrier (SLC) members has not been elucidated in colorectal cancer (CRC). We aimed to identify a prognostic signature from the SLC members and comprehensively analyze their roles in CRC. Firstly, we downloaded transcriptome data and clinical information of CRC samples from GEO (GSE39582) and TCGA as training and testing dataset, respectively. We extracted the expression matrix of SLC genes and established a prognostic model by univariate and multivariate Cox regression. Afterwards, the low-risk and high-risk group were identified. Then, the differences of prognosis traits, transcriptome features, clinical characteristics, immune infiltration and drug sensitivity between the two groups were explored. Furthermore, molecular subtyping was also implemented by non-negative matrix factorization (NMF). Finally, we studied the expression of the screened SLC genes in CRC tumor tissues and normal tissues as well as investigated the role of SLC12A2 by loss of function and gain of function. As a result, we developed a prognostic risk model based on the screened 6-SLC genes (SLC39A8, SLC2A3, SLC39A13, SLC35B1, SLC4A3, SLC12A2). Both in the training and testing sets, CRC patients in the high-risk group had the poorer prognosis and were in the more advanced pathological stage. What's more, the high-risk group were enriched with CRC progression signatures and immune infiltration. Two groups showed different drug sensitivity. On the other hand, two distinct subclasses (C1 and C2) were identified based on the 6 SLC genes. CRC patients in the high-risk group and C1 subtype had a worse prognosis. Furthermore, we found and validated that SLC12A2 was steadily upregulated in CRC. A loss-of-function study showed that knockdown of SLC12A2 expression restrained proliferation and stemness of CRC cells while a gain-of-function study showed the contrary results. Hence, we provided a 6-SLC gene signature for prognosis prediction of CRC patients. At the same time, we identified that SLC12A2 could promote tumor progression in CRC, which may serve as a potential therapeutic target.

Comprehensive analysis of the relationship between cuproptosis-related gene GCSH and prognosis, tumor microenvironment infiltration, and therapy response in endometrial cancer.

BACKGROUND: Cuproptosis, a novel form of cell death regulated by protein lipoylation and implicated in mitochondrial metabolism. However, the impact of the cuproptosis-related gene γ-glutamylcysteine synthetase (GCSH) on endometrial cancer (EC) prognosis, tumor immune microenvironment, and therapeutic response remains to be further researched. METHODS: The differential expression of GCSH between endometrial cancer and normal tissues was analyzed using multiple public databases. Additionally, cancer and adjacent tissues were prospectively collected from 17 EC patients, and immunohistochemical analysis was performed to further investigate GCSH expression differences. The relationship between GCSH and the prognosis and clinicopathological characteristics of EC patients was evaluated, and a nomogram was constructed to predict patient survival based on GCSH expression. Then, Gene set variation analysis (GSVA) was utilized to explore the potential biological functions of GCSH in EC. The impact of GCSH on the tumor microenvironment (TME) was estimated. Finally, the effect of GCSH on the response to immunotherapy and chemotherapeutic drugs in EC was investigated. RESULTS: The expression of GCSH was significantly upregulated in EC. High GCSH expression was associated with poor prognosis in EC patients. Enrichment analysis showed that high GCSH was associated with immune suppression. Furthermore, high GCSH was found to be associated with a non-inflamed TME, leading to decreased infiltration levels of immune cells. Finally, it was observed that patients with high GCSH were insensitive to both immunotherapy and chemotherapeutic drugs. CONCLUSION: This study revealed the role of GCSH in TME, response to therapy, and clinical prognosis in EC, which provided novel insights for the therapeutic application in EC.

Guanylate-binding proteins signature predicts favorable prognosis, immune-hot microenvironment, and immunotherapy response in hepatocellular carcinoma.

BACKGROUND: The role of guanylate-binding proteins (GBPs) in various cancers has been elucidated recently. However, our knowledge of the clinical relevance and biological characteristics of GBPs in hepatocellular carcinoma (HCC) remains limited. METHODS: A total of 955 HCC patients were enrolled from five independent public HCC cohorts. The role of GBP molecules in HCC was preliminarily investigated, and a GBP family signature, termed GBPs-score, was constructed by principal component analysis to combine the GBP molecule values. We revealed the effects of GBP genes and GBPs-score in HCC via well-established bioinformatics methods and validated GBP1-5 experimentally in a tissue microarray (TMA) cohort. RESULTS: GBPs molecules were closely associated with the prognosis of patients with HCC, and a high GBPs-score highly inferred a favorable survival outcome. We also revealed high GBPs-score was related to anti-tumor immunity, the immune-hot tumor microenvironment (TME), and immunotherapy response. Among the GBPs members, GBP1-5 rather than GBP6/7 may be dominant in these fields. The TMA analysis based on immunohistochemistry showed positive correlations between GBP1-5 and the immune-hot TME with abundant infiltration of CD8+ T cells in HCC. CONCLUSIONS: Our integrative study revealed the genetic and immunologic characterizations of GBPs in HCC and highlighted their potential values as promising biomarkers for prognosis and immunotherapy.

Identification and validation of immune-associated NETosis subtypes and biomarkers in anti-neutrophil cytoplasmic antibody associated glomerulonephritis.

Background: NETosis is a new form of cell death, marked by DNA chromatin release from dead neutrophils. While it aids in microbe defense, it may worsen inflammation in autoimmune diseases, causing tissue harm. The impact of NETosis on Anti-neutrophil Cytoplasmic Antibody-associated Glomerulonephritis (ANCA-GN) remains unexplored and requires investigation. Methods: First, a weighted gene co-expression network analysis (WGCNA) was conducted to uncover differential expression of neutrophil extranuclear trap-associated genes (DE-NETs) in ANCA-GN. The NETosisScore model was established through the single sample gene set enrichment analysis (ssGSEA), which categorized all patients into high-risk and low-risk groups. The accuracy of model was assessed by ROC curve. The biological function of various subgroups was explored through Gene Set Variation Analysis (GSVA), while the abundance of immune cell infiltration was measured with CIBERSORT. Furthermore, the key NETosis-related genes (NRGs) were identified using three machine learning algorithms, and their relationship with renal function was analyzed through the NephroseqV5 database. Through the application of qPCR and immunohistochemical staining techniques, the mRNA and protein expression levels of NRGs were determined in patients with ANCA-GN and control. Results: A NETosisScore model was developed from 18 DE-NETs using the ssGSEA algorithm. The model's ability to predict ANCA-GN patients with a ROC AUC of 0.921. The high-risk group in ANCA-GN showed enrichment of immune-related pathways and greater infiltration of immune cells, as revealed by KEGG enrichment analysis and CIBERSORT. Using three machine learning algorithms, we identified six NRGs. Significant positive correlations were found between NRGs and CCR, macrophages, T-cell co-inhibition, and TIL. Further KEGG analysis revealed that the functions of NRGs may be closely related to the toll-like receptor signaling pathway. The levels of NRGs increased as kidney function declined and were positively correlated with Scr (serum creatinine) and negatively correlated with GFR (glomerular filtration rate), qPCR analysis showed increased expression of most NRGs in ANCA-GN patients. Furthermore, immunohistochemical staining confirmed higher expression of all NRGs in ANCA-GN patients. Conclusion: NETosisScore model accurately predicts high-risk patients in ANCA-GN with enriched immune pathways, 6 NRGs identified as potential biomarkers.

Twenty versus 40 back-and-forth needle movements for endoscopic ultrasound-guided fine-needle biopsy of solid pancreatic masses: a prospective, crossover, randomized study.

BACKGROUND AND AIMS: In endoscopic ultrasound (EUS)-fine-needle biopsy (FNB) of solid pancreatic mass lesions, the number of times the needle moves back and forth within the lesion might affect the collection of the sample and the subsequent diagnostic accuracy. Thus, this study was designed to compare the diagnostic adequacy between different numbers of back-and-forth movements in EUS-FNB. METHODS: Fifty-five patients with solid pancreatic masses underwent EUS-FNB sampling with the needle (22-gauge) moved 20 times (MTT) and 40 times (MFT) randomly and sequentially for a total of four alternating passes. We compared the acquisition rate of appropriate and adequate specimens for histologic assessment and diagnostic accuracy. RESULTS: Finally, 55 patients (35 men and 20 women) were included in the study. We found that 56.4% (31/55) and 60% (33/55) of the specimens obtained using MTT and MFT, respectively, could be adequately diagnosed histologically (P = 0.815, McNemar test). The diagnostic accuracy of MTT and MFT was 72.7% (40/55) and 80% (44/55), respectively (P = 0.289, McNemar test). The overall diagnostic accuracy was 89.1%. CONCLUSION: There was no significant statistical difference between the histopathological diagnostic samples obtained in MTT and those obtained in MFT. Therefore, a large number of back-and-forth movements of the needle should be avoided during EUS-FNB, which can help reduce the operation time and may reduce the risk of intraoperative and postoperative complications (Clinical trial registration number: ChiCTR2000031106).

Novel oviduct endoscope combining optical coherence tomography with intratubal ultrasonography for fallopian tube exploration: An in vivo rabbit pilot study.

BACKGROUND AND STUDY AIM: Currently, several limitations exist in the examination of the oviduct. In this study, the usefulness and feasibility of a novel ultrafine dual-modality oviduct endoscopy device for in vivo assessment of the oviduct were evaluated. METHODS: Five Japanese white rabbits were selected to undergo oviduct probing using a combination of optical coherence tomography (OCT) and intratubal ultrasonography. The feasibility of the procedure was evaluated through 152 pairs of clear, clinically interpretable images obtained using spiral scanning via the pull-back method. OCT images were compared with the oviduct histopathology sections. RESULTS: Visualization of the oviduct using both OCT and ultrasound revealed a differentiated three-layer tissue; however, ultrasound showed a poorer clarity than OCT. By comparing OCT images with the histological morphology of the oviduct, the inner low-reflective layer of the oviduct corresponds to the mucosal layer, the middle high-reflective layer corresponds to the fibrous muscle layer, and the outer low-reflective layer corresponds to the connective tissue layer. Postoperatively, the general condition of the animals was good. CONCLUSION: This study demonstrated the feasibility and potential clinical value of the novel ultrafine dual-modality oviduct endoscope. Dual-modality imaging of OCT and intratubal ultrasonography can provide clearer microstructure of the oviduct wall.

Esophagogastroduodenoscopy Outcomes Variated by the Time of the Day: A Single-Center Experience.

(1) Background: To assess whether the start time influences the outcomes of esophagogastroduodenoscopy (EGD). (2) Methods: We retrospectively analyzed the clinical data of patients who underwent EGD between January 2021 and December 2021 in our endoscopy center. The EGD were divided into three shifts, according to the start time. The lesion detection rate (LDR) and endoscopy biopsy rate (EBR) were used to evaluate the quality of the EGD. (3) Results: A total of 14,597 procedures were included in this study. The LDR of shift 2 was significantly lower than that of shift 1 (62.4% vs. 58.5%; p < 0.001). The EBR of shift 1 (37.4% vs. 31.5%; p < 0.001) and shift 3 (35.5% vs. 31.5%; p = 0.024) were significantly higher than that of shift 2; the EBR in shift 1 did not differ significantly from shift 3 (p = 0.280). The multivariable analysis for the EGD performed before 14:00 demonstrated a graded decrease in the LDR and EBR after adjusting the confounders (p < 0.001). (4) Conclusion: In a continuous working period, the lesion detection and biopsy submission of EGD are superior to those in the first three hours compared to the last three hours; the LDR and EBR decreased as the day progressed, probably due to the endoscopists' fatigue.

Comprehensive analysis of cuproptosis-related genes in prognosis, tumor microenvironment infiltration, and immunotherapy response in gastric cancer.

BACKGROUNDS: Cuproptosis is the most recently identified copper-dependent cell death form that influences tricarboxylic acid (TCA) cycle. However, the relationship between cuproptosis and clinical prognosis, tumor microenvironment infiltration (TME), and response to immunotherapy remains unclear. METHODS: Single-sample gene-set enrichment analysis (ssGSEA) was employed to construct cuproptosisScore (cpS) and 1378 gastric cancer (GC) patients from five independent public datasets were classified into high- or low-cpS groups according to the median of cpS. Then the impacts of cuproptosis on tumor microenvironment infiltration (TME), biological function, response to immunotherapy, and clinical prognosis of GC were evaluated. RiskScore and nomogram were constructed using Lasso Cox regression algorithm to validate its predictive capability in GC patients. RESULTS: Compared to patients with high cpS, patients with low cpS exhibited poorer prognosis, higher TNM stage, and stronger stromal activation. Meanwhile, the analysis of response to immunotherapy confirmed patients with high cpS could better benefit from immunotherapy and had a better susceptibility to chemotherapeutic drugs. Then, 9 prognosis-related signatures were collected based on differentially expressed genes (DEGs) of cpS groups. Finally, a riskScore model was constructed using the multivariate Cox (multi-Cox) regression coefficients of prognosis-related signatures and had an excellent capability of predicting 1-, 3-, and 5-year survival in GC patients. CONCLUSIONS: This study revealed the role of curproptosis in TME, response to immunotherapy, and clinical prognosis in GC, which highlighted the significant clinical implications of curproptosis and provided novel ideas for the therapeutic application of cuproptosis in GC.

A deep learning network based on multi-scale and attention for the diagnosis of chronic atrophic gastritis.

BACKGROUND AND STUDY AIM: Chronic atrophic gastritis plays an important role in the process of gastric cancer. Deep learning is gradually introduced in the medical field, and how to better apply a convolutional neural network (CNN) to the diagnosis of chronic atrophic gastritis remains a research hotspot. This study was designed to improve the performance of CNN on diagnosing chronic atrophic gastritis by constructing and evaluating a network structure based on the characteristics of gastroscopic images. METHODS: Three endoscopists reviewed the endoscopic images of the gastric antrum from the Gastroscopy Image Database of Zhongnan Hospital and labelled available images according to pathological results. Two novel modules proposed recently were introduced to construct the Multi-scale with Attention net (MWA-net) considering the characters of similar medical images. After training the network using images of training sets, the diagnostic ability of the MWA-net was evaluated by comparing it with those of other deep learning models and endoscopists with varying degrees of expertise. RESULTS: As a result, 5,159 images of the gastric antrum from 2,240 patients were used to train and test the MWA-net. Compared with the direct application of famous networks, the MWA-net achieved the best performance (accuracy, 92.13%) with an increase of 1.80% compared to that of ResNet. The suspicious lesions indicated by the network are consistent with the conclusion of experts. The sensitivity and specificity of the convolutional network for gastric atrophy diagnosis are 90.19% and 94.51%, respectively, which are higher than those of experts. CONCLUSIONS: Highly similar images of chronic atrophic gastritis can be identified by the proposed MWA-net, which has a better performance than other well-known networks. This work can further reduce the workload of gastroscopists, simplify the diagnostic process and provide medical assistance to more residents.

Novel biliopancreatic duct endoscope combining optical coherence tomography with intraductal US for exploring the bile duct: a diagnostic study in a porcine model.

BACKGROUND AND AIMS: Existing biliopancreatic duct endoscopy is deficient in the examination of early biliary and pancreatic tumors. This study aimed to evaluate the usefulness and feasibility of a novel ultrafine, separable, biliopancreatic duct endoscopy device with dual modalities of intraductal US (IDUS) and optical coherence tomography (OCT) for the in vivo assessment of the biliopancreatic duct system during ERCP. METHODS: Five Bama miniature pigs were selected to probe their common bile duct and branches by using this novel equipment during ERCP. The feasibility of the procedure was evaluated by clear, clinically interpretable images obtained by using spiral scanning with the pull-back method. The clinical usefulness of the novel product was evaluated by postoperative choledochoscopy and assessment of the animal's general condition. RESULTS: One hundred forty-one pairs of images from 5 Bama miniature pigs were acquired. Visualization of the bile duct using both OCT and IDUS was characterized by a differentiated 3-layer architecture, whereas IDUS had poor clarity when compared with OCT. Postoperative choledochoscopy showed no local lesion in the bile duct wall, and the general condition of animals was normal. CONCLUSIONS: This prospective evaluation indicated the feasibility and potential clinical value of the novel, ultrafine, separable biliopancreatic duct endoscopy device. The fusion of the 2 imaging modalities can shed light on the early diagnosis of biliary and pancreatic tumors. Further studies will be carried out to establish diagnosis criteria with the dual-modality imaging using an animal pathologic model and a human clinical study.