Cholangiocarcinoma (CCA) is a hepatobiliary carcinoma with uncontrolled cell proliferation, poor prognosis, and high mortality. The ovarian tumor structural domain (OTU) containing protein 6B (OTUD6B) belongs to the OTU deubiquitin family and is vital in tumor development. However, its expression and biological function in CCA remain unknown. The expression of OTUD6B in CCA was analyzed using TIMER2.0, UALCAN, and GEO databases. MTT, clonal formation assay, immunofluorescence staining, immunohistochemistry staining, and flow cytometry examined the regulation of OTUD6B on cell proliferation, cycle, and apoptosis. The effects of OTUD6B on tumor volume and weight were assessed using the xenograft tumor model. The activities of PTK2 and STAT3 were detected by western blot and CO-IP. The biological database identified that OTUD6B was upregulated in CCA. In CCA cells, OTUD6B knockdown reduced CCA cell proliferation and promoted apoptosis. Cell cycle analysis indicated that the cycle stopped at the G0/G1 phase after OTU6B downregulation. Furthermore, OTUD6B knockdown resulted in a decrease in tumor volume and weight in xenograft tumor models. Mechanistically, OTUD6B is involved in the deubiquitination of PTK2. PTK2 further affected the phosphorylation of STAT3 thereby regulating the CCA process. Our study demonstrates that OTUD6B knockdown participates in the ubiquitination of PTK2 and phosphorylation of STAT3 to alleviate the process of CCA. These results suggest that OTUD6B may be a potential new strategy for CCA treatment.

The Ten-Eleven Translocation (TET) family genes are implicated in a wide array of biological functions across various human cancers. Nonetheless, there is a scarcity of studies that comprehensively analyze the correlation between TET family members and the molecular phenotypes and clinical characteristics of different cancers. Leveraging updated public databases and employing several bioinformatics analysis methods, we assessed the expression levels, somatic variations, methylation levels, and prognostic values of TET family genes. Additionally, we explored the association between the expression of TET family genes and pathway activity, tumor microenvironment (TME), stemness score, immune subtype, clinical staging, and drug sensitivity in pan-cancer. Molecular biology and cytology experiments were conducted to validate the potential role of TET3 in tumor progression. Each TET family gene displayed distinct expression patterns across at least ten detected tumors. The frequency of Single Nucleotide Variant (SNV) in TET genes was found to be 91.24%, primarily comprising missense mutation types, with the main types of copy number variant (CNV) being heterozygous amplifications and deletions. TET1 gene exhibited high methylation levels, whereas TET2 and TET3 genes displayed hypomethylation in most cancers, which correlated closely with patient prognosis. Pathway activity analysis revealed the involvement of TET family genes in multiple signaling pathways, including cell cycle, apoptosis, DNA damage response, hormone AR, PI3K/AKT, and RTK. Furthermore, the expression levels of TET family genes were shown to impact the clinical staging of tumor patients, modulate the sensitivity of chemotherapy drugs, and thereby influence patient prognosis by participating in the regulation of the tumor microenvironment, cellular stemness potential, and immune subtype. Notably, TET3 was identified to promote cancer progression across various tumors, and its silencing was found to inhibit tumor malignancy and enhance chemotherapy sensitivity. These findings shed light on the role of TET family genes in cancer progression and offer insights for further research on TET3 as a potential therapeutic target for pan-cancer.

Objective: To clarify the potential therapeutic effects of thymoquinone (TQ) on pancreatic cancer and its gemcitabine (GEM) sensitivity. Methods: The expression levels of hypoxia inducible factor-1α (HIF-1α), collagens (COL1A1, COL3A1, and COL5A1), and transforming growth factor-β1 (TGFβ1) in pancreatic cancer and para-carcinoma tissues were compared using immunohistochemical methods, and their relationships with TNM staging were analyzed. The effects of TQ on apoptosis, migration, invasion, and GEM sensitivity of pancreatic cancer cells were assessed using in vitro and in vivo experiments. Western blot and immunohistochemistry were used to detect the expression levels of HIF-1α, extracellular matrix (ECM) production pathway-related proteins, and TGFβ/Smad signaling pathway-related proteins. Results: The expression levels of HIF-1α, COL1A1, COL3A1, COL5A1, and TGFβ1 in pancreatic cancer tissues were significantly higher than those in para-carcinoma tissues and correlated with TNM staging ( p < 0.05). TQ and GEM administration inhibited the migration and invasion of the human pancreatic cancer cell line PANC-1 and promoted the apoptosis of PANC-1 cells. The combination of TQ and GEM was more effective than GEM alone. Western blot analysis showed that the expression levels of HIF-1α, ECM production pathway-related proteins, and TGFβ/Smad signaling pathway-related proteins were significantly decreased when TQ was used to treat PANC-1 cells ( p < 0.05), and the expression levels of these proteins in the TQ + GEM group were significantly more decreased than those in the GEM group. Overexpression or knockdown of HIF-1α in PANC-1 cells showed the same effects as those induced by TQ administration. In vivo experiments showed that in PANC-1 tumor-bearing mice, tumor volume and tumor weight in mice treated with GEM and TQ were significantly lower than those in control or GEM-treated mice, whereas cell apoptosis was significantly increased ( p < 0.05). Western blot and immunohistochemistry results showed that the levels of HIF-1α, ECM production pathway-related proteins, and TGFβ/Smad signaling pathway-related proteins in the GEM + TQ treatment group were further decreased compared to the control group or the GEM treatment group ( p < 0.05). Conclusion: In pancreatic cancer cells, TQ can promote apoptosis, inhibit migration, invasion, and metastasis, and enhance the sensitivity to GEM. The underlying mechanism may involve the regulation of ECM production through the TGFβ/Smad pathway, in which HIF-1α plays a key role.

Background and Aim Refractory anastomotic leakage (RAL) after intersphincteric resection (ISR) usually leads to failure of protective stoma reversal in ultralow rectal cancers. The aim of this study is to assess the risk factors and oncological outcomes of both anastomotic leakage (AL) and RAL, and quality of life (QoL) of RAL after laparoscopic ISR (LsISR).Methods A total of 371 ultralow rectal cancer patients with LsISR were enrolled from a tertiary colorectal surgery referral center. Risk factors for AL and RAL were identified by logistic regression. Three-year disease-free survival (DFS) of AL and RAL was analyzed by the Cox regression. QoL of RAL group (compared with non-RAL group) was assessed using the European Organization for Research and Treatment of Cancer QLQ-C30 and QLQ-CR29 questionnaires.Results The rates of AL and RAL after LsISR accounted for 8.4% (31/371) and 4.6% (17/371) in this cohort, respectively. Non-left colic artery preservation (odds ratio [OR] = 3.491, P = 0.009), neoadjuvant chemoradiotherapy (nCRT) (OR = 6.038, P < 0.001), and lower anastomosis height (OR = 5.271, P = 0.010) were independent risk factors for AL, while nCRT (OR = 11.602, P < 0.001) was the only independent risk factor for RAL. Male (hazard ratio [HR] = 1.989, P = 0.014), age > 60 years (HR = 1.877, P = 0.018), and lymph node metastasis (HR = 2.125, P = 0.005) were independent risk factors of poor 3-year DFS, but not RAL (P = 0.646). RAL patients have significantly worse global health status, worse emotional and social function scores at the late postoperative stage, and worse urinary and sexual function at the early postoperative stage (all P < 0.05).Conclusions Neoadjuvant chemoradiotherapy was an independent risk factor for RAL after LsISR. RAL shows similar oncological outcomes, but with poor QoL.

Breast cancer (BC) is a highly prevalent aggressive malignancy in women worldwide, and the search for key targets in its pathogenesis is a focus of research. Long non-coding RNAs (lncRNAs) play an important role in many cancers, including breast cancer. This study aimed to investigate the role of lncRNA SNHG9 in BC. The expression of SNHG9 in BC cells was found to be higher than that of human mammary epithelium. SNHG9 was found to inhibit the proliferation, migration, and invasion of cells and promote apoptosis. It was also found that SNHG9 regulates the miR-326/Wnt5a/β-catenin axis to promote the development of BC. Dual luciferase reporter and RNA pull-down assays confirmed the interaction between SNHG9, Wnt5a, and miR-326. Western blot analysis indicated that the expression of Wnt5a, β-catenin, c-myc, and cyclin D1 decreased significantly after the silencing of SNHG9 and the overexpression of miR-326. On the contrary, Wnt5a, β-catenin, c-myc, and cyclin D1 proteins were significantly up-regulated after inhibiting miR-326 expression. These findings suggest that SNHG9 is a promising target for BC therapy.

Background: Intratumoral hypoxia is widely associated with the development of malignancy, treatment resistance, and worse prognoses. The global influence of hypoxia-related genes (HRGs) on prognostic significance, tumor microenvironment characteristics, and therapeutic response is unclear in patients with non-small cell lung cancer (NSCLC).Method: RNA-seq and clinical data for NSCLC patients were derived from The Cancer Genome Atlas (TCGA) database, and a group of HRGs was obtained from the MSigDB. The differentially expressed HRGs were determined using the limma package; prognostic HRGs were identified via univariate Cox regression. Using the least absolute shrinkage and selection operator (LASSO) and multivariate Cox regression, an optimized prognostic model consisting of nine HRGs was constructed. The prognostic model’s capacity was evaluated by Kaplan‒Meier survival curve analysis and receiver operating characteristic (ROC) curve analysis in the TCGA (training set) and GEO (validation set) cohorts. Moreover, a potential biological pathway and immune infiltration differences were explained.Results: A prognostic model containing nine HRGs (STC2, ALDOA, MIF, LDHA, EXT1, PGM2, ENO3, INHA, and RORA) was developed. NSCLC patients were separated into two risk categories according to the risk score generated by the hypoxia model. The model-based risk score had better predictive power than the clinicopathological method. Patients in the high-risk category had poor recurrence-free survival in the TCGA (HR: 1.426; 95% CI: 0.997–2.042; p = 0.046) and GEO (HR: 2.4; 95% CI: 1.7–3.2; p < 0.0001) cohorts. The overall survival of the high-risk category was also inferior to that of the low-risk category in the TCGA (HR: 1.8; 95% CI: 1.5–2.2; p < 0.0001) and GEO (HR: 1.8; 95% CI: 1.4–2.3; p < 0.0001) cohorts. Additionally, we discovered a notable distinction in the enrichment of immune-related pathways, immune cell abundance, and immune checkpoint gene expression between the two subcategories.Conclusion: The proposed 9-HRG signature is a promising indicator for predicting NSCLC patient prognosis and may be potentially applicable in checkpoint therapy efficiency prediction.

Abstract Purpose To clarify the therapeutic effects of thymoquinone (TQ) in pancreatic cancer (PC) and the potential hypoxia-inducible factor-1α (HIF-1α)-mediated mechanisms. Methods Western blotting and polymerase chain reaction were used to detect the expression levels of HIF-1α in PC cells treated with TQ. The effects of TQ on proliferation, migration, invasion, cell cycle, and apoptosis of PC cells and pancreatic ductal epithelial cells were assessed using in vitro experiments. Results TQ significantly inhibited the migration and invasion ability of PANC-1 cells (p < 0.05) but had no effects in hTERT-HPNE cells, and it significantly increased the proportion of the G2 phase and promoted apoptosis in PANC-1 cells compared with those in hTERT-HPNE cells (p < 0.05). Additionally, TQ significantly inhibited the expression of HIF-1α and phosphatidyl inositol-4,5-bisphosphate-3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/Akt/mTOR)-related proteins in PANC-1 cells (p < 0.05). TQ affected the interaction between heat shock protein 90 (HSP90) and HIF-1α in PANC-1 cells. Conclusion TQ showed suppressive effects on proliferation, migration, invasion, cell cycle, and apoptosis of PC; promoted ubiquitination-mediated degradation of HIF-1α by affecting the interaction of HIF-1α with HSP90; and reduced HIF-1α synthesis by affecting PI3K/Akt/mTOR signaling pathway. The findings suggest that TQ might have potential anti-tumor effects in PC.

目的 探讨腹腔镜直肠癌超低位前切除术(Ls-uLAR)并发直肠吻合口漏病人的转归,并对造口回纳后再发吻合口漏的危险因素进行初步分析.方法 回顾性分析北京大学第一医院普通外科2012年1月至2020年12月同一手术团队完成的Ls-uLAR并发吻合口漏的31例直肠癌病人的临床资料及随访结果,对造口回纳后再发吻合口漏的临床特征及危险因素进行分析.结果 371例接受Ls-uLAR的病人中有31例(8.4％)术后并发吻合口漏.其中预防性造口术后吻合口漏23例(23/307,7.5％),其转归为:1例围手术期死亡,1例吻合口复发再次行经腹-会阴联合切除术,6例吻合口重度狭窄难以回纳,15例经保守治疗吻合口漏临床愈合(其中2例因肺转移而放弃回纳);无预防性造口术后吻合口漏8例(8/64,12.5％),其转归为:1例围手术期死亡,2例急诊行回肠造口,5例经保守治疗愈合.符合吻合口漏临床愈合标准的15例病人于初次术后3～16个月行造口回纳,其中8例(8/15,53.3％)再次出现吻合口漏.单因素分析结果显示:新辅助放化疗(是vs.否:100.0％ vs.30.0％,P=0.026)、初次术中出血量(＞ 50 mL vs.≤50 mL 87.5％ vs.14.3％,P=0.010)及吻合口内镜下缺血征象(有vs.无:85.7％ vs.25.0％,P=0.041)与再发吻合口漏相关.8例再发吻合口漏病人表现为骶前脓肿并继发不完全性肠梗阻4例,反复发作肛周脓肿和肛瘘2例,直肠阴道瘘2例.所有再发漏病人经保守治疗1～2个月均未能愈合,除1例直肠阴道瘘拒绝再次造口外,其余7例均改行横结肠造口.结论 腹腔镜直肠癌超低位前切除术并发吻合口漏结局不良,继发吻合口狭窄及回纳后再发吻合口漏的风险较高,对吻合口漏病人的临床愈合标准、造口回纳时机和手术方式,尤其是新辅助放化疗后病人仍有待进一步研究.

Pathogenesis of hepatic Echinococcosis is induced by parasitism of Echinococcus species.Generally speaking, there are two types of the disease--hepatic cystic echinococcosis which is caused by E. granulosus and hepatic alveolar echinococcosis which is caused by E. multilocularis.Either of the two types is much clinically popular.However, simultaneous appearance of both the two types can be rarely seen.In this study, we reported a case of co-infection of both hepatic cystic and alveolar echinococcosis, aiming to offer experience in diagnosing and treating the co-infection by depicting the clinical characteristics, imaging manifestations and therapies.

Objectives: To examine the risk factors of anastomotic leakage for low rectal cancers undergoing laparoscopic intersphincteric resection (ISR), and to construct a nomogram prediction model for it. Methods: The perioperative data of 302 low rectal cancer patients undergoing laparoscopic ISR by the same surgical team of Department of General Surgery, Peking University First Hospital between January 2012 and January 2019 were retrospectively reviewed. There were 190 males and 112 females, aging 60(14) years (range: 20 to 84 years). χ2 test, independent sample t test, U test and Logistic regression analysis were used to analyze the risk factors for anastomotic leakage. R software was used to complete the drawing of the nomogram prediction model, and the receiver operating characteristic curve was used to evaluate the predictive ability of the nomogram prediction model. Results: There were 24 patients (7.9%) had anastomotic leakage among the 302 patients enrolled, including 10 cases of grade A leakage, 9 cases of grade B leakage, and 5 cases of grade C leakage. Out of the 24 patients, 2 patients (8.3%) died, 3 patients (12.5%) received leakage-related reoperation. Median healing time of the anastomotic leakage was 74 (58) days (range: 14 to 180 days). Univariate analysis showed male gender (P=0.009), preoperative serum albumin concentration (P=0.004), neoadjuvant radiochemotherapy (P=0.017), preserving left colonic artery (P=0.002) and performing a diverting ileostomy (P=0.015) were significantly correlated with anastomotic leakage. Logistic multivariate analysis showed male gender (OR=6.052, 95%CI: 1.535 to 23.860, P=0.010), neoadjuvant radiochemotherapy (OR=4.098, 95%CI: 1.318 to 12.821, P=0.015), no preserving left colonic artery (OR=16.699, 95%CI: 3.051 to 91.406, P=0.001) and not performing a diverting ileostomy (OR=21.218, 95%CI: 4.341 to 103.710, P<0.01) were independent risk factors for anastomotic leakage. According to the results of multi-factor regression analysis, the nomogram prediction model was constructed. The area under the curve of the nomogram prediction model was 0.840 (95%CI: 0.766 to 0.914). After internal verification, the concordance index value of the model was 0.840. Conclusion: Male gender, neoadjuvant radiochemotherapy, no preserving left colonic artery and not performing a diverting ileostomy are independent risk factors for anastomotic leakage for low rectal cancers undergoing laparoscopic ISR.

Background Intracorporeal rectal transection at the anorectal junction for ultralow rectal cancer is technically difficult due to pelvic width and limited roticulation, which might require a transanal transection or an oblique transection with multiple firings. These procedures were reported to be associated with the increased risk of morbidity. To address these problems, we presented a novel technique Transanterior Obturator Nerve Gateway (TANG) to transect rectum for ultralow rectal cancer and evaluated its safety and feasibility in this study. Methods A total of 210 consecutive patients who underwent laparoscopic coloanal anastomosis with or without partial intersphincteric resection (CAA/pISR) for rectal cancers between January 2017 and January 2020 were included. Eighty of these patients were analyzed using propensity score matching (PSM). The perioperative characteristics, TANG-related variables, and genitourinary and anal function outcomes were analyzed. Results Among these enrolled patients, 170 patients underwent traditional transection, and 40 underwent TANG transection; the patients were matched to include 40 patients in each group by PSM. After PSM, there were no significant differences in the operating time ( p = 0.351) or bleeding volume ( p = 0.474) between the two groups. However, the TANG group had fewer cases of conversion to transanal transection (0 vs. 13, p < 0.001). Moreover, the patients in TANG group had a more desirable transection with longer distal resection margin (1.7 vs. 1.1 cm, p < 0.001), shorter stapling line (6.6 vs. 10.3 cm, p < 0.001) and fewer stapler firings ( p < 0.001). The overall postoperative complication rates and genitourinary and anal function outcomes were not significantly different between the two groups. Conclusions The TANG approach appears to be a safe, feasible and effective approach for intracorporeal ultralow rectal transection with more distal resection, more vertical transection and fewer stapler firings.

Background Circulating microRNAs that post-transcriptionally regulate gene expressions have been reported as promising biomarkers in cancer monitoring. This study was to identify the potential role of circulating miR-212 in gastric cancer and whether it could serve as a novel biomarker for gastric cancer. Methods We detected the serum levels of miR-212 in 100 health people and 110 gastric cancer patients and analyzed the relationships of the serum level of miR-212 with gastric cancer. We detected the expression of miR-212 in human gastric mucosal epithelial cell line (GES-1) and human gastric cancer cell lines (NCI-N87 and SNU-16) using qRT-PCR. Then, we detected the role of 5-aza-deoxycytidine on the epigenetic regulation of miR-212 in human gastric cancer cell lines. Furthermore, luciferase reporter assay was used to detect binding activity of miR-212 on SOX4 mRNA, and their functions on the cell proliferation and apoptosis. Results The expression of miR-212 was higher in health people than that in gastric cancer patients, higher in gastric mucosal epithelial cell line than that in gastric cancer cells. miR-212 can be a circulating biomarker and an independent prognostic factor of gastric cancer. Moreover, miR-212 can directly regulate the 3 ' UTR of SOX4 mRNA to suppress p53 and Bax, resulting gastric cancer cells proliferation inhibition and apoptosis induction. Conclusion Our study demonstrated that miR-212 was epigenetically downregulated in gastric cancer, and resulting low level of miR-212 can be a potential circulating biomarker and poor prognosis predicator of gastric cancer.

目的 评价腹腔镜后盆腔脏器切除术治疗局部进展期直肠癌的安全性及近期疗效,分析腹腔镜后盆腔脏器切除术操作要点.方法 回顾性分析2014年4月至2019年10月由同一术者行腹腔镜后盆腔脏器切除术的15例局部进展期直肠癌患者临床资料,包括手术相关指标、病理指标和随访情况.结果 15例患者均顺利完成腹腔镜后盆腔脏器切除术(LPPE),无中转开腹病例.消化道重建方式中9例为结肠—直肠/肛管吻合,3例为乙状结肠造瘘,3例为远端直肠封闭近端乙状结肠造瘘;阴道处理方式中12例为阴道残端缝合,3例为阴道后壁切除.3例术前MRI检查提示侧方淋巴结肿大而行单侧侧方淋巴结清扫;1例联合膀胱部分切除,1例联合输尿管部分切除,1例联合右半结肠切除.平均手术时间(268.3±80.9)min,中位出血量150(100,200)mL,平均术后住院时间(15.2±7.2)d,2例患者出现术后并发症,1例为阴道残端瘘,保守治疗3周后痊愈出院,1例为直肠吻合口漏继发直肠阴道瘘,后期遗留吻合口狭窄,至今保护性回肠造口未能还纳.全组无术后膀胱功能恢复延迟.所有手术均为R0切除,11例为癌性侵犯,8例伴区域淋巴结转移.中位随访时间2年,随访期间4例患者因肿瘤术后复发转移死亡,11例患者实现无病生存,中位生存时间25个月.结论 腹腔镜后盆腔脏器切除术治疗局部进展期直肠癌安全可行,近期疗效尚可,远期肿瘤学效果仍需进一步扩大样本进行研究.

Background Castleman disease (CD) is a rare lymphoproliferative disease characterized by high heterogeneity in clinical manifestation and prognosis. This study aimed to summarize clinical features of localized retroperitoneal CD and our experiences to improve the diagnosis and treatment of this disease. Methods Clinical data of 45 patients with localized retroperitoneal CD were retrospectively analyzed. The differences in clinical features between groups with and without paraneoplastic pemphigus (PNP) were compared. Survival was analyzed between groups depending on whether complicating with PNP, bronchiolitis obliterans (BO), gender, age and uni-centric CD (UCD)/multi-centric CD (MCD), respectively. Results Significant differences were observed between patient groups in the prevalence of retroperitoneal CD with PNP complicated with BO (P=0.010), the constituent ratios of initial symptoms (P<0.001) and the duration from appearance of the initial symptoms to being diagnosed (P=0.009). Among 45 cases, 43 tumors had clear margins and intact envelops and were completely resected, 40 patients were cured or significantly relieved, 3 patients were not significantly relieved, 2 patients received palliative surgical therapy and eventually relapsed and died after surgery. There were significant differences in the survival rate between groups depending on complication with BO, gender and age (≤40 and >40 years) (all P<0.05). Conclusions Prompt and complete removal of the retroperitoneal CD tumor is critical to the management of this disease, as palliative resection tends to cause relapse and lead to a poor prognosis. Retroperitoneal CD patients with PNP may develop complications from BO leading to death. Complication with PNP, complication with BO, male gender and age ≥40 years were identified as prognostic risk factors for patients with localized retroperitoneal CD.

Background Gastric cancer (GC) is one of the most common and lethal malignancies worldwide. Therefore, a better understanding of the mechanism of its malignant progression and chemoresistance will be helpful for the treatment of patients with GC. Methods The gene expression profiles downloaded from GEO database and the TargetScan Human were used to identify the key regulation model based on miRNA by bioinformatics analyses. The regulation of miRNA to target was clarified by luciferase assay, qPCR, and Western blotting. Then, the in vitro and in vivo experiments were further conducted by overexpression or knockdown of miRNA and/or target to examine the regulation effects and clarify the mechanism. Results In the present study, miR-424-3p was identified to be differentially expressed among normal gastric, GC, and chemoresistant GC tissues. Target analysis results indicated that ABCC2, a chemoresistance-related gene, was a regulated target of miR-424-3p. The in vitro and in vivo experiment results further demonstrated that miR-424-3p relied on ABCC2-induced chemoresistance to promote GC proliferation and metastasis. Conclusion Overall, this study revealed that miR-424-3p contributed to the malignant progression and chemoresistance of GC. Thus, miR-424-3p could be a potential target for the treatment of GC.

BACKGROUND:Gastric cancer is a common malignant tumor in the clinic with a high mortality rate, ranking the first among malignant tumors of the digestive system. Early gastric cancer exhibits no specific clinical symptoms and signs, and most of the patients were diagnosed as advanced gastric cancer. The prognosis is poor, and the 5-year overall survival rate is still lower than 30%, seriously threatening people's life and health. However, the pathogenesis of gastric cancer is still unclear.METHODS:This study aimed to identify methylated differentially expressed genes in gastric cancer and to study the cellular functions and pathways that may be involved in its regulation, as well as the biological functions of key methylated differentially expressed genes. The gene expression data set and methylation data set of gastric cancer genes based on TCGA were analyzed to identify prognostic methylated genes.RESULTS:This study showed that the methylation of the DERL3 promoter was correlated with the clinical analysis of tumors. Further studies were conducted on genes co-expressed with DERL3, whose functions and pathways to inhibit gastric cancer were adaptive immune response, T cell activation, immune response-regulating pathway, cell surface on molecules, and natural killer cell-mediated cytotoxicity. Finally, cell proliferation assay, cell scratch assay, and cell invasion assay confirmed that DERL3 as a tumor suppressor gene inhibited the malignant evolution of gastric cancer.CONCLUSIONS:The analysis of key methylated differentially expressed genes helped elucidate the epigenetic regulation mechanism in the development of gastric cancer. DERL3, as a methylation biomarker, has a predictive and prognostic value in the accurate diagnosis and treatment of gastric cancer and provides potential targets for the precision treatment of gastric cancer.TRIAL REGISTRATION:Not applicable.

Echinococcosis is a disease caused by the Echinococcus species that parasitizes in humans. Alveolar echinococcosis (AE) which is caused by Echinococcus multilocularis is harmful to humans. AE mainly occurs in the liver and can be transferred to retroperitoneal lymph nodes, lung, brain, bone, spleen and other organs through lymphatic and blood vessels. Cholangiocarcinoma can occur in the intrahepatic and extrahepatic bile ducts and is more common in the hilar. We reported a case of hilar bile duct alveolar echinococcosis which was originally misdiagnosed an cholangiocarcinoma.

TF/FVIIa (Tissue Factor/Active Coagulation factor VII) and EGFR (Epidermal Growth Factor Receptor) signaling both promote malignant progression of colorectal cancer. However, the crosstalk of these two signaling pathways in human colorectal cancer cells remains unclear. Here we detected the changes of mRNA profile in human colorectal cancer cell SW620 exposed to FVIIa. Microarray showed that mRNA levels of EGFR ligands were significantly upregulated. Western blot analysis confirmed the upregulation of EGFR ligands and the phosphorylation of EGFR at tyrosine-845 in colorectal cancer cells exposed to FVIIa. However, knockdown of TF by RNAi could block the upregulation of EGFR ligands induced by FVIIa stimulation. On the other hand, the expression of components of TF/FVIIa signaling was significantly upregulated in LoVo cells stimulated by EGF. However, the crosstalk between the two signaling pathways could not be detected in HT-29 colon cancer cells bearing wild-type KRAS. Taken together, our study suggest that the crosstalk between TF/FVIIa and EGFR signaling pathways in colon cancer cells depends on KRAS mutation.

Objective To detect the expression profile changes of human colon cancer cell SW620 when its tissue factor/active coagulation factor Ⅶ (TF/FⅦa) pathway is activated and discover new downstream effectors of TF/FⅦa pathway.Methods Total RNAs of two groups of SW620 cells (treated with 100 nmol/L FⅦa for 24 h in treament group and identical volume of phosphate-buffered saline in control group) were extracted and hybridization was performed on gene chips.Differentially expressed genes were screened and part of the gene expressions were verified using real-time reverse transcriptase-polymerase chain reaction (RT-qPCR),network of differentially expressed genes and pathways were analysed.Gene expressions of epidermal growth factor receptor (EGFR) ligands amphiregulin (AREG),epiregulin (EREG),Heparin-binding EGF-like growth factor (HB-EGF),transforming growth factor-α (TGF-α),β-cellulin (BTC),epidermal growth factor (EGF) and epigen (EPGN) were detected by RT-qPCR following activation of TF/FⅦa pathway of SW620,SW480,LoVo and HCT116 cells.Results There were 263 differentially expressed genes with a test/control fold change of ≥ 1.5 (150 upregulated,113 downregulated).Network analysis showed that genes of laminin-integrin system,pathways of glycometabolism and cancer related pathways were associated with TF/F Ⅶ a signalling's activation.Gene expressions of AREG,EREG,HB-EGF and TGF-α of SW620 were upregulated in a time-dependent manner and reached peaks at 12 h,relative gene expressions of control group vs treatment group were 0.92±0.13 vs.5.50 ±0.91(P=0.000),1.00±0.15 vs.2.17±0.44(P=0.002),1.11 ±0.08 vs.1.73 ±0.32(P =0.005),0.94 ±0.12 vs.1.49 ±0.09(P =0.002).When TF/FⅦa pathway was activated for 6 h,in SW480 cells,gene expressions of AREG and BTC were significantly upregulated,relative expressions were t.00 ±0.25 vs.2.19 ±0.60(P =0.033),1.00 ±0.23 vs.4.41 ±0.56 (P =0.001),expression of EPGN was not detected,in LoVo cells,AREG and BTC were significantly upregulated and EGF was significantly downregulated:1.00 ± 0.19 vs.1.87 ± 0.39 (P =0.025),1.00 ± 0.01 vs.1.86±0.06 (P=0.000),1.00±0.08 vs.0.68 ±0.16 (P=0.035),in HCT116 cells,AREG,EREG,HB-EGF,TGF-α,and EGF were significantly upregulated:1.00 ± 0.09 vs.1.38 ± 0.06 (P=0.004),1.00±0.02 vs.1.14±0.03 (P=0.003),1.00±0.03 vs.1.85 ±0.11 (P=0.000),1.00±0.05 vs.1.26±0.10 (P=0.015),1.00±0.26 vs.1.91 ±0.22 (P=0.010).Conclusion In SW620 cells,activation of TF/FⅦa pathway can induce expression changes of genes of laminin-integrin system,pathways of glycometabolism and cancer related pathways and EGFR pathway,these effectors and pathways may take critical roles in TF/FⅦa pathway-related bioeffect.

OBJECTIVE:To preliminarily verify the cross talk between tissue factor/active coagulation factor VII (TF/FVIIa) and epidermal growth factor receptor (EGFR) pathways in human colon cancer cells in culture.METHODS:FVIIa was treated to HT-29 (KRAS-wild type) and LoVo (KRAS-mutant) colon cancer cells to activate TF/FVIIa pathway, qRT-PCR and Western blot were used to detect the expressions of amphiregulin (AREG) and epiregulin (EREG), ligands of EGFR on mRNA and protein levels, respectively. After knocking down expression of TF by TF-targeted siRNA transfection, FVIIa was treated and mRNA expressions of AREG and EREG were detected to see whether the FVIIa-induced effects were dependent on TF. Expressions of mRNA of TF and FVII were detected by qRT-PCR following the activation of EGFR pathway by treatment with epidermal growth factor (EGF) to HT-29 and LoVo cells.RESULTS:After TF/FVIIa pathway was activated, for HT-29 cells, expressions of AREG (on mRNA level) and EREG (both on mRNA and protein level) were significantly down-regulated versus those of control group, gene expressions of AREG and EREG were 0.55±0.09 vs.0.99 ±0.09, 0.67±0.10 vs.1.02±0.02, protein expressions of EREG were 0.54±0.09 vs.1.04±0.13, all P<0.05. For LoVo cells, expressions of AREG (both on mRNA and protein level) and EREG (on protein level) were significantly up-regulated versus those of control group, gene expression of AREG were 1.87±0.39 vs. 0.93±0.23, protein expressions of AREG and EREG were 3.09±0.73 vs.1.11±0.21, 1.53±0.19 vs.0.97±0.23, all P<0.05. The regulating effect of AREG and EREG mRNA expression by FVIIa in HT-29 and LoVo cells could both be partly blocked by knocking down TF expression. For HT-29 cells, activation of EGFR pathway induced no significant TF mRNA expression, FVII mRNA expression was not detected. However,for LoVo cells, activation of EGFR pathway induced significantly higher mRNA expressions of both TF and FVII, expressions were 1.53±0.23 vs.1.00±0.23, 53.20±6.08 vs.1.00±0.15, all P<0.05.CONCLUSION:In colon cancer cell LoVo, when activated, TF/FVIIa pathway and EGFR pathway could interact through upregulating the other pathway's effectors, and mutant KRAS might play a critical role in the two pathways' cross talk.