BACKGROUND:Approximately 50% of colorectal cancer (CRC) patients exhibit high levels of Fusobacterium nucleatum (Fn), which is associated with chemotherapy resistance. As Fn itself cannot be directly targeted for therapy in vivo, elucidating the mechanism underlying Fn-induced chemotherapy resistance is crucial, though it remains unclear. METHODS:qPCR was used to analyze the correlation between Fn abundance and clinical parameters in 80 human colon cancer samples. The effect of Fn on the sensitivity of colon cancer cells to oxaliplatin was evaluated by clone formation, EdU proliferation and apoptosis assays. RNA sequencing was performed on Fn-infected colon cancer cells to identify differentially expressed genes. Mechanistic studies explored the interaction between PVT1 and ATAD3A, and the role of TLR4/NF-κB pathway in regulating PVT1 expression. RESULTS:qPCR experiments showed that Fn abundance was associated with later clinical stage and shorter RFS. Clone formation, EdU proliferation assay and apoptosis assay showed that Fn could change the sensitivity of colon cancer cells to oxaliplatin. Further RNA sequencing showed that Fn infection could upregulate the expression of long noncoding RNA plasmacytoma variant translocation 1 (PVT1) in colon cancer cells. The abundance of Fn in colon cancer tissues was positively correlated with the level of PVT1, and the mechanism was that PVT1 binds to AAA domain protein 3 (ATAD3A) and prevents its ubiquitination. Fn upregulates ATAD3A expression through PVT1 and inhibits ER stress-mediated cell death. In addition, in colon cancer cells co-cultured with Fn, PVT1 expression is regulated by the TLR4/NF-κB pathway. CONCLUSIONS:This study delineates a pathway where Fn infection promotes oxaliplatin resistancein colon cancer cells by upregulating PVT1 via the TLR4/NF-κB pathway. PVT1 subsequently stabilizes ATAD3A, suppressing cell death. PVT1 is a potential target to overcome the high abundance of Fusobacterium nucleatum leading to oxaliplatin resistance in colon cancer.

This retrospective analysis aimed to assess the feasibility and safety of endoscopic retrograde cholangiopancreatography (ERCP) in pediatric patients by examining ERCP-related adverse events (AEs) occurring over a decade at a single center. Pediatric patients under 18 years old who underwent ERCP at the Second Hospital of Hebei Medical University from 1/2013 to 11/2023 were included. ERCP-related AEs were defined according to ERCP-related adverse events: European Society of Gastrointestinal Endoscopy (ESGE) Guideline. Clinical data of patients experiencing ERCP-related AEs were obtained from electronic medical records for analysis. Over the past decade, a total of 76 pediatric patients underwent 113 ERCP procedures, including 26 patients who underwent repeat ERCP, totaling 63 procedures. There were 32 males and 44 females, with a median age of 13 years (range 3 years and 5 months–17 years and 9 months). Among all ERCP procedures, 14 (12.4

Nonischemic cardiomyopathy (NICM) is a major cause of advanced heart failure, and the morbidity and mortality associated with NICM are serious medical problems. However, the etiology of NICM is complex and the related mechanisms involved in its pathogenesis remain unclear. The microarray datasets GSE1869 and GSE9128 retrieved from the Gene Expression Omnibus database were used to identify differentially expressed genes (DEGs) between NICM and normal samples. The co-expressed genes were identified using Venn diagrams. Kyoto Encyclopedia of Genes and Genomes pathway analyses and gene ontology enrichment were used to clarify biological functions and signaling pathways. Analysis of protein-protein interaction networks using Search Tool for the Retrieval of Interacting Genes/Proteins online to define the hub genes associated with NICM pathogenesis. A total of 297 DEGs were identified from GSE1869, 261 of which were upregulated genes and 36 were downregulated genes. A total of 360 DEGs were identified from GSE9128, 243 of which were upregulated genes and 117 were downregulated genes. In the 2 datasets, the screening identified 36 co-expressed DEGs. Kyoto Encyclopedia of Genes and Genomes pathway and gene ontology analysis showed that DEGs were mainly enriched in pantothenate and CoA biosynthesis, beta-alanine metabolism, kinetochore, G-protein beta/gamma-subunit complex, and other related pathways. The PPI network analysis revealed that DUSP6, EGR1, ZEB2, and XPO1 are the 4 hub genes of interest in the 2 datasets. Bioinformatics analysis of hub genes and key signaling pathways is an effective way to elucidate the mechanisms involved in the development of NICM. The results will facilitate further studies on the pathogenesis and therapeutic targets of NICM.

Increased glycolysis is one of the key metabolic hallmarks of cancer cells. However, the roles of lncRNAs in energy metabolism and cancer metastasis remain unclear. Here, the expression of TMEM105 associated with glycolysis was dramatically elevated from normal to breast cancer to breast cancer liver metastasis tissues, and the survival analysis revealed that high TMEM105 expression was related to poor survival, especially in patients with liver metastasis. Moreover, TMEM105 facilitated the glycolysis of breast cancer cells and induced cell invasion and breast cancer liver metastasis (BCLM). Mechanistically, TMEM105 regulated LDHA expression by sponging miR-1208, which further promoted cell glycolysis and BCLM. Importantly, glycolytic production of lactate enhanced TMEM105 expression in breast cancer cells by activating the SHH-MAZ signaling pathway. These findings suggested that the lactate-responsive TMEM105 acted as a miRNA sponge, inducing BCLM via a glycolysis-mediated positive feedback loop, which might be a rational target for the treatment of BCLM patients.

Purpose: Long noncoding RNAs (lncRNAs) are correlated with cancer pathogenesis and prognosis. Many studies have shown that aberrant expression of MIR31HG is implicated in the cancer progression and patient prognosis. However, the biological function and predictive value of MIR31HG in colorectal cancer is unclear. Methods: The correlation between MIR31HG expression and clinicopathological characteristics of colorectal cancer patients was analyzed by collating the information from The Cancer Genome Atlas (TCGA) database. Kaplan-Meier analysis, univariable and multivariable Cox regression analysis were performed to evaluate the prognostic value of MIR31HG. Gene set enrichment analysis (GSEA) was conducted to identify the potential carcinogenic mechanisms implicated in MIR31HG. Moreover, MIR31HG was knocked down using siRNA in colorectal cancer cells, and cell migration, invasion, growth and colony formation assays were performed. The expression of MIR31HG influenced gene markers was quantified by qRT-PCR in MIR31HG-silenced colorectal cancer cells. Results: In TCGA database, we found that MIR31HG was elevated in colorectal cancer patients. The patients with high MIR31HG expression had poor overall survival and disease-specific survival. Univariable and multivariable analyses showed that MIR31HG expression was an independent prognostic predictor in colorectal cancer patients. GSEA revealed that MIR31HG mainly modulated focal adhesion, extracellular matrix organization, integrin cell surface interactions and focal adhesion-PI3K-Akt-mTOR-signaling pathway. Besides, MIR31HG knockdown significantly impaired colorectal cancer cell migration, invasion, growth and colony formation. Further qRT-PCR data confirmed that alteration of MIR31HG expression notably affected the tumorigenesis-related key gene expression in the cells. Conclusion: Our findings provide evidence that MIR31HG is a key factor in maintaining the malignant phenotype of colorectal cancer and act as an independent predictor for patients with colorectal cancer.

Background:Undifferentiated pleomorphic sarcoma (UPS) is a malignant tumor that originates in the mesenchymal tissue and is common in the extremities and retroperitoneum. Primary UPS of the duodenal papilla is rare and a distinct clinical entity.Case presentation:In this report, a 48-year-old Chinese man was admitted to our hospital with symptoms of melena. The patient underwent choledochectomy and choledochaljejunostomy for obstructive jaundice 8 years before admission. Endoscopic examination after admission confirmed a mass located at the duodenal papilla. Then, the duodenal papilla and tumor resection were performed, and the histopathology report confirmed the diagnosis of UPS. The patient refused further treatment and died 2 months later due to local recurrence and intrahepatic metastasis.Conclusions:It is rare that the mass in the duodenal papilla is diagnosed as UPS. The unpredicted behavior of these tumors warrants a careful plan considering their indolent nature and possible recurrence and metastasis. The prognosis was poor despite the early complete resection.

Background:Cholangiocarcinoma is a primary malignant tumor, and its progression involves oncogene activation, the absence of tumor suppressor gene, abnormal signaling pathways and miRNA expression. MiRNAs are abnormally ex pressed in many types of tumors. Objective:This study aims to observe the effects of miR-582 on cholangiocarcinoma cell proliferation, S-phase arrest, migration and invasion and to analyze the regulation of miR-582 on LIS1 to clarify the real role of miR-582 in cholangiocarcinoma development. Materials and Methods: TCGA database of cholangiocarcinoma samples was analyzed. Dual fluorescence reporter and TargetScan were conducted to confirm whether LIS1 was the target gene of miR-582. Effects of miR-582 and LIS1 on HCC-9810 cell proliferation, S-phase cell ratio, migration and invasion were determined by CCK-8, Flow cytometry and Transvvell, respectively, whereas the function of iniR-582 on MMP-2 and P-Akt expression was identified by Western blotting. Nude mice xenograft model of cholangiocarcinoma was established to detect what miR-582 did for tumor growth. Results:TCGA showed that miR-582 was lowly expressed and LIS1 was highly expressed in donor tissues compared with adjacent tissues. MiR-582 targeted LIS1 to inhibit MMP-2 and p-AKT expression. Transfection of miR-582 mimics could suppress HCC-9810 cell proliferation, S-stage arrest, migration and invasion, while LIS1 worked oppositely. MiR-582 inhibitors promoted cell biological behavior, whereas LIS1 siRNA was opposite. In nude mice xenograft model, miR-582 overexpression inhibited tumor growth. Conclusions:It implies that miR-582 could negatively regulate LIS1 to inhibit MMP-2 and P-Akt expression, thus suppressing cell invasion and proliferation in cholangiocarcinoma.

目的 对外科完成经内镜胰胆管造影术(ERCP)规范化培训后的独立术者成长期所有病例进行回顾性研究,分析术后并发症发生情况,并绘制其学习曲线.方法 确定ERCP操作过程中的可量化评价标准包括乳头插管时间、手术并发症(急性胰腺炎、出血、穿孔、急性胆管炎)发生率、尝试插管次数等六项,采集自2018年7月至2020年12月全部的ERCP手术操作记录中以上参数的具体值进行统计分析,学习曲线评价指标计算公式为:δ=Xi-X0,计算每一次操作的6个评价指标的累积和值即该次ERCP操作水平量化值的总和(∑=δ1+δ2+δ3+δ4+δ5+δ6),量化后绘制学习曲线并分析学习期术后并发症发生率.结果 该术者总体插管成功率为99.4％,高于一般文献报道的独立ERCP操作术者的插管成功率要求.取前100例ERCP操作病例绘制CUSUM控制图并拟合学习曲线.拟合曲线公式:y=10.7001+0.1020x+0.008372x2-0.000089x3,决定系数R2=0.899,曲线拟合效果较好.68例ERCP手术后,曲线k值为负,故68例ERCP手术后,该术者跨越学习曲线.结论 外科术者即使没有内镜基础直接进行ERCP操作培训完全可以胜任ERCP手术,其插管成功率及手术并发症发生情况较文献报道的无明显升高.

Hypoxia induces a series of cellular adaptive responses that enable promotion of inflammation and cancer development. Hypoxia-inducible factor-1α (HIF-1α) is involved in the hypoxia response and cancer promotion, and it accumulates in hypoxia and is degraded under normoxic conditions. Here we identify prostate cancer associated transcript-1 (PCAT-1) as a hypoxia-inducible long non-coding RNA (lncRNA) that regulates HIF-1α stability, crucial for cancer progression. Extensive analyses of clinical data indicate that PCAT-1 is elevated in breast cancer patients and is associated with pathological grade, tumor size, and poor clinical outcomes. Through gain- and loss-of-function experiments, we find that PCAT-1 promotes hypoxia-associated breast cancer progression including growth, migration, invasion, colony formation, and metabolic regulation. Mechanistically, PCAT-1 directly interacts with the receptor of activated protein C kinase-1 (RACK1) protein and prevents RACK1 from binding to HIF-1α, thus protecting HIF-1α from RACK1-induced oxygen-independent degradation. These findings provide new insight into lncRNA-mediated mechanisms for HIF-1α stability and suggest a novel role of PCAT-1 as a potential therapeutic target for breast cancer.

BackgroundLncRNAs have proven to be involved in the initiation and progression of cholangiocarcinoma (CCA), although the mechanism by which this occurs remains unknown.MethodsThe current study reveals that RHPN1-AS1 was overexpressed in CCA patient samples, which predicted poor outcome of CCA patients. RHPN1-AS1 increased in vitro pancreatic carcinoma cell proliferation as well as promoted xenograft growth in vivo. Mechanistically, DANCR upregulated expression of YAP1 by competitively binding to miR-345-5p. Importantly, RHPN1-AS1 level was positively correlated with YAP1 expression level in CCA tissues. Moreover, YAP1 overexpression could predicted a poor outcome of CCA patients.ResultsTaken together, our results suggested that RHPN1-AS1 might be a remarkable biomarker to evaluate prognosis in CCA.ConclusionThe RHPN1-AS1/YAP1 axis may provide new strategies for CCA clinical practice.

Objective:To analyze the relationship between the expression of microRNA (miRNA, miR)-150 and epithelial-mesenchymal transition (EMT) in gallbladder cancer and its mechanism.Methods:The expression of miR-150 in 30 cases of gallbladder cancer was detected by real-time quantitative polymerase chain reaction (qPCR), and 30 cases of chronic cholecystitis tissues was used as control. Simultaneously, the expression of proliferating cell nuclear antigen (PCNA), E-cadherin, N-cadherin and matrix metalloproteinase (MMP)-9 was detected, and the relationship of miR-150 and these genes was analyzed. Human gallbladder cancer cell line GBC-SD was transfected with miR-150 mimics. Then scratch experiments and Transwell chamber experiments were used to detect changes in cell migration and invasion; qPCR and Western blotting were used to detect changes in mRNA and protein of each gene.Results:The expressions of miR-150 and E-cadherin in gallbladder cancer tissues were lower than those in chronic cholecystitis tissues ( t=-7.035, -14.146, P<0.01). The expressions of PCNA, N-cadherin and MMP-9 were significantly higher compared with chronic cholecystitis tissues ( t=3.813, 9.339, 5.616, P<0.01). The expression of miR-150 was related to the degree of tumor differentiation, clinical stage, and lymph node metastasis ( t=3.228, 2.396, -2.604, P<0.05). Pearson correlation analysis showed that miR-150 was positively correlated with E-cadherin ( r=0.630, P<0.05), and negatively correlated with PCNA, N-cadherin, and MMP-9 ( r=-0.769, -0.628, -0.616, P<0.05). After transfection, the migration and invasion ability of GBC-SD cells decreased ( F=1 965.860, 114.940, P<0.01). The expressions of PCNA, N-cadherin, and MMP-9 were decreased, while the expression of E-cadherin was increased ( P<0.05). Conclusion:Decreased miR-150 expression in gallbladder cancer plays a key role in tumor progression, and the mechanism may be that miR-150 participates in tumor EMT process by regulating some genes.

The authors have requested that this preprint be withdrawn due to author disagreement.

Background Outcomes of gastroesophageal reflux disease (GERD) using Toupet fundoplication (TF) and Stretta radiofrequency (SRF) have not been compared and this study was conducted to compare therapeutic efficacy of the two methods. Methods This retrospective study analyzed a total of 230 patients undergoing TF or SRF at our hospital. Baseline data, reflux symptoms, the DeMeester scores, lower esophageal sphincter (LES) pressure and adverse events were compared over 1 year period. Results A total of 226 patients were included in the study. The time and frequency of reflux and percentage of reflux time before and 12 months after therapy were not significantly different. There were significantly interactions between the therapy method and follow-up time on the DeMeester score and LES pressure. Twelve months post therapy, the DeMeester score was significantly higher in SRF than in TF group, while the LES pressure was lower. At 12 months after therapy, multivariate Cox proportional regression analysis showed that reflux frequency, the DeMeester score and LES pressure were risk factors for poor prognosis in TF group, while reflux frequency and the DeMeester score, and LES pressure were risk factors for poor prognosis in SFR group. Conclusions Compared with TF, SFR can significantly improve the esophageal pH and pressure in GERD patients without increasing the risk of poor prognosis.

LncRNAs have been reported to be involved in the initiation and progression of cholangiocarcinoma (CCA), although the mechanisms by which this occurs remains unknown. The current study aimed to reveal the clinico-pathological relationship of RHPN1-AS1 expressed with CCA, and also investigate the functions of RHPN1-AS1 both in vivo and in vitro. RHPN1-AS1 was overexpressed in CCA cell proliferation as well as promoted xenograft growth in vivo. Mechanistically, DANCR upregulated the expression of YAP1 by competitively binding to miR-345-5p. Importantly, RHPN1-AS1 level was positively correlated with YAP1 expression level in CCA tissues (P<0.05). Moreover, Kaplan-Meier curves showed that YAP1 overexpression predicted a poor outcome of CCA patients (P=0.004). Taken together, our results suggested that RHPN1-AS1 might be a remarkable biomarker to evaluate prognosis in CCA. The RHPN1-AS1/YAP1 axis may provide new strategies for CCA clinical practice.

Abstract The authors have requested that this preprint be removed from Research Square.

目前胆管癌仍是胆道系统最常见的恶性肿瘤[1?2] ,其起病隐匿,患者就诊的首要症状多为渐进性黄疸,发现时往往已属晚期,失去了手术切除机会,根治性手术五年生存率仅10%,预后极差[3] .对于无法行根治性手术或者不愿意行根治性手术的患者,内镜下胆道引流术可以有效解除阻塞性胆汁淤积,减轻黄疸?皮肤瘙痒?腹痛等相关并发症,预防胆管炎并避免渐进性胆道梗阻引起的肝衰竭等,是解除恶性胆道梗阻的一种有效的姑息性治疗手段.

BACKGROUND:A variety of factors contribute to biliary injury that is difficult to be repaired. Stent implantation is extensively used for bile duct injury, but either scaffolds made by metal or plastics can lead to certain adverse reactions. OBJECTIVE:To explore the biological characteristics of a novel biodegradable scaffold and its repair effects on bile duct injury. METHODS:The biological characteristics of the novel biodegradable scaffold were detected by fresh bile, and its degradation was observed at different time points. Thirty Bama mini pigs were included and were randomly divided into observation group (n=15) and control group (n=15). After bile duct injury models were prepared, the control group was subjected to the bile duct interrupted suture, while the observation group was subjected to the novel biodegradable scaffold combined with omentum majus. The biological properties of the scaffolds were observed. Hepatic enzymes and serum total bilirubin levels were detected, as wel as hematoxylin-eosin staining, Masson staining and immunohistochemistry detection ofα-smooth muscle actin were performed. RESULTS AND CONCLUSION:Before and 1, 3 and 6 months after surgery, hepatic enzymes and total bilirubin of two groups were detected, and neither intra-group nor intergroup comparisons had significant differences (P>0.05). Hematoxylin-eosin staining and Masson staining revealed that inflammatory reactions and fiber hyperplasia at the anastomotic site in the observation group were lighter than those in the control group at different time points after surgery. Theα-smooth muscle actin-positive scores in both two groups were in a rise at 1 and 3 months after surgery, and peaked at the 3rd month, and then began to decline. Moreover, theα-smooth muscle actin-positive scores in the observation group were significantly lower than those in the control group at 3 and 6 months after surgery (P<0.05). These results show that the novel biodegradable scaffold has good biological characteristics and can obtain ideal repair effects in the bile duct injury.

measured to compare the correlation between the changes of T lymphocyte subsets before and after operation and the change of platelet. Results Thirty-one patients were underwent laparoscopic splenectomy. The T lymphocyte subsets and CD3 + ,CD4 + and CD3 + / CD4 + in all patients showed an increase trend as the time prolonged after operation,and the differences were significant( F = 6. 91,3. 93,4. 18;P = 0. 023,0. 004,0. 011). While the ratio of CD3 + / CD8 + and CD3 + CD4 + / CD3 + CD8 + were decreased as time increased,and differences were statistically significant( F = 2. 59,3. 67;P = 0. 001,0. 002). They were related with platelet change after operation and correlation statistics analysis showed R values were 0. 332,0. 271,0. 345,- 0. 119,- 0. 164,and the P value were 0. 039,0. 021,0. 002,0. 017,0. 023. Conclusion ITP patients have the imbalance of T lymphocyte subsets,and splenectomy can improve the imbalance of T lymphocyte subsets in patients with ITP. T lymphocyte subsets may serve as a reference index to indicate the prognosis of the patients.