Figure S2: IL-18 receptor (IL-18R) expression in pancreatic cancer tissues and cell lines.

Figure S4: IL-18 promotes PC cell proliferation, invasion and metastasis.

Figure S5: IL-18 promotes PC cell proliferation, invasion and metastasis.

Figure S6: IL-18 promotes proliferation and invasion of PC cells via the NF-κB pathway.

We investigated the role of actin filament associated protein 1 antisense RNA1 (AFAP1-AS1) lncRNA in promoting cholangiocarcinoma (CCA). qRT-PCR analysis of patient samples showed that AFAP1-AS1 expression was higher in CCA tumors than matched adjacent non-tumor tissue. AFAP1-AS1 levels were also higher in CCA cell lines (HuCCT1 and TFK-1) than a normal biliary epithelium cell line (HIBEpic). AFAP1-AS1 knockdown in CCA cell lines using shAFAP1-AS1 reduced cell proliferation and colony formation in CCK-8 and colony formation assays, respectively. Cell cycle analysis demonstrated that AFAP1-AS1 knockdown resulted in G0/G1 cell cycle arrest and inhibition of S-G2/M transition compared to the controls. CCA cells transfected with shAFAP1-AS1 also exhibited reduced metastasis and invasiveness in Transwell and wound healing assays. This was further confirmed in xenograft experiments with nude mice using CCA cells transfected with shAFAP1-AS1 or control shRNA. AFAP1-AS1 knockdown cells produced smaller tumors, demonstrating that AFAP1-AS1 promotes tumor growth in vivo. AFAP1-AS1 knockdown also increased expression of actin filament associated protein 1 (AFAP1) and reduced cell stress filament integrity, as determined from western blot and immunofluorescence assays, respectively. These findings indicate that AFAP1-AS1 exerts oncogenic effects in CCA. We postulate that AFAP1-AS1 is a potentially useful diagnostic and prognostic biomarker and therapeutic target for CCA.

Background: Validity of the laparoscopic approach in pancreatic head lesion remains debatable. This study aims to compare the safety and effectiveness of laparoscopic pancreatoduodenectomy (LPD) and open pancreatoduodenectomy (OPD) and investigate the source of heterogeneity from surgeons' and patients' perspectives. Method: We searched PubMed, Cochrane, Embase, and Web of Science for studies published before February 1, 2021. Of 6578 articles, 81 were full-text reviewed. The primary outcome was mortality. Three independent reviewers screened and extracted the data and resolved disagreements by consensus. Studies were evaluated for quality using ROB2.0 and ROBINS-I. According to different study designs, sensitivity and meta-regression analyses were conducted to explore the heterogeneity source. This meta-analyses was also conducted to explore the learning curve's heterogeneity. This study was registered with PROSPERO, CRD42021234579. Results: We analyzed 34 studies involving 46,729 patients (4705 LPD and 42,024 OPD). LPD was associated with lower (P = 0.025) in unmatched studies (P = 0.017). No differences in mortality existed in randomized controlled trials (P = 0.854) and matched studies (P = 0.726). Sensitivity analysis found no significant difference in mortality in elderly patients, patients with pancreatic cancer, and in high- and low-volume hospitals (all P > 0.05). In studies at the early period of LPD (<40 cases), higher mortality (P < 0.001) was found (all P < 0.05). LPD showed non-inferiority in length of stay, complications, and survival outcomes in all analyses. Conclusion: In high-volume centers with adequate surgical experience, LPD in selected patients appears to be a valid alternative to LPD with comparable mortality, LOS, complications, and survival outcomes.

[This corrects the article DOI: 10.18632/oncotarget.16880.].

BACKGROUND:Delayed gastric emptying (DGE) is a common complication following laparoscopic pancreaticoduodenectomy (LPD), although it remains incompletely understood, and only few studies have investigated the clinical benefits of hepatic branch of the vagus nerve (HBVN) preservation on DGE after LPD until now. We intended to evaluate the effect of preservation of the HBVN during LPD on the incidence of DGE.METHODS:A total of 274 consecutive LPDs performed at a single center between July 2014 and December 2019 with available videos were retrospectively reviewed. DGE was defined according to the International Study Group of Pancreatic Surgery (ISGPS) criteria, and HBVN condition during the LPD procedure was evaluated through a video review. Risk factors associated with DGE were assessed by performing univariate and multivariate logistic regression analyses. Postoperative outcomes between the HBVN-preserved and HBVN-injury groups were compared before and after propensity score matching (PSM).RESULTS:One hundred fifty-six (56.93%) patients underwent LPD with HBVN-preserved and 118 (43.07%) with HBVN injury. DGE occurred in 33.2% of patients (n = 91) with grades B and C occurring at 13.9% (n = 38) and 7.7% (n = 21), respectively. Longer operative time, more EIBL, HBVN injury, POPF (grades B and C), postoperative hemorrhage, intra-abdominal infection, and Clavien-Dindo ≥III were identified as risk factors for DGE in the univariate analysis. Then, in the multivariate analysis, HBVN injury and intra-abdominal infection were found to be independent risk factors affecting the incidence of DGE (any grade) or clinically relevant DGE (grades B and C). Furthermore, the prevalence of DGE was significantly higher in the HBVN-injury group than in the HBVN-preserved group before and after PSM analysis (46.61% vs. 23.08%, P<0.001; 42.59% vs. 23.15%, P=0.013).CONCLUSIONS:HBVN preservation during LPD might be associated with a reduced incidence of DGE as a framework for prospective quality improvement.

目的 探讨百里醌(T Q)对大鼠肝脏缺血再灌注损伤(IRI)的保护作用及分子机制.方法 建立大鼠肝脏IRI模型,肝脏缺血再灌注前,实验大鼠进行百里醌给药预处理.通过酶联免疫吸附试验(ELISA)测定血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)水平,检测肝脏组织谷胱甘肽过氧化物酶(GPX)、谷胱甘肽(GSH)、超氧化物歧化酶(SOD)、丙二醛(MDA)等酶活性;蛋白印迹法(Western blotting)检测肝脏组织凋亡相关蛋白.实验设3组:假手术组(Sham组,n=10)、缺血再灌注组(IRI组,n=10)、百里醌预处理+肝缺血再灌注组(TQ+IRI组,各亚组n=10).结果 与Sham组比较,IRI组大鼠血清ALT、AST水平显著增高,TQ+IRI组ALT、AST较IRI组明显降低(P<0.05).IRI组与Sham组比较,大鼠肝脏组织中GPX、GSH、SOD含量均明显下降(P<0.05),而MDA含量显著增高(P<0.05);TQ+IRI组与IRI组比较,GPX、GSH、SOD含量均显著上升(P<0.05),MDA含量下降(P<0.05).百里醌预处理能下调肝脏IRI的Bax、裂解的胱天蛋白酶(Cleaved-caspase)-3、Cleaved-caspase-9表达,阻断肝细胞凋亡.结论 百里醌可能通过调控肝细胞氧化应激诱导的细胞凋亡进而在肝脏IRI中发挥肝脏保护作用.

目的:探讨携带人mda-7/IL-24基因的溶瘤腺病毒SG600-IL24对肝癌细胞选择性杀伤效应的机制.方法:构建携带人mda-7/IL-24基因的溶瘤腺病毒SG600-IL24后,用其分别感染肝癌细胞株HepG2、HCCLM3和正常肝细胞株L02.用RT-PCR和Western blot分别检测感染后,各细胞株STAT3以及其信号通路下游相关信号分子基因与蛋白的表达变化,以及磷酸化STAT3(p-STAT3)蛋白的表达变化.结果:成功构建携带人mda-7/IL-24基因的溶瘤腺病毒SG600-IL24载体.SG600-IL24感染后,3种细胞的mda-7/IL-24基因及蛋白表达均明显升高(均P＜0.05).两种肝癌细胞感染后,STAT3的表达水平明显下调,其下游信号分子c-myc、Bcl-xl、Bcl-2、cyclin D2、survivin、MMP-2、MMP-9、XIAP、OPN、VEGF的表达水平明显下调,Bax表达明显上调,均呈一定的时间依赖趋势(均P＜0.05);p-STAT3蛋白在感染后上升,并在感染2h达到峰值,随后下降.感染后,L02细胞中未见STAT3及其下游信号分子表达的变化(均P＞0.05).结论:携带人mda-7/IL-24基因溶瘤腺病毒SG600-IL24选择性杀伤肝癌细胞的机制可能与其选择性抑制肝癌细胞STAT3信号通路,而对正常肝细胞不产生明显影响.

目的 探讨Oddi括约肌松弛合并胆总管结石复发的手术治疗方案.方法 胆总管复发结石合并Oddi括约肌松弛342例(均行数次胆道手术,胆囊均已切除),根据手术治疗方式不同分为三组,Ⅰ组281例,行横断胆管+胆肠Roux-en-Y吻合术,Ⅱ组40例,行开腹或腔镜胆总管切开取石+T管引流术,Ⅲ组21例,行ERCP术,比较3种手术方式的治疗效果.结果 Ⅰ组结石复发率、胆管炎发生率均低于Ⅱ组和Ⅲ组(P＜0.05),复发平均时间则晚于Ⅱ组和Ⅲ组(P＜0.05),而Ⅱ组和Ⅲ组结石复发率、胆管炎发生率及复发平均时间比较差异无统计学意义(P＞0.05).胆肠Roux-en-Y吻合术的治疗总有效率高于胆总管切开取石+T管引流术和ERCP术(P＜0.05),而胆总管切开取石术治疗总有效率与ERCP术比较差异无统计学意义(P＞0.05).结论 横断胆管+胆肠Roux-en-Y吻合术是目前治疗Oddi括约肌松弛合并复发胆管结石较有效的手术方案.

目的:探究miR-486-5p在肝细胞性肝癌(HCC)中的变化及其调节HCC细胞系增殖的可能机制.方法:q-PCR法检测98例健康人血浆和87例HCC患者组织与血浆miR-486-Sp含量,并采用Pearson相关性分析其与甲胎蛋白(AFP)升高的相关性,同时检测人HCC癌细胞系SMMC-7721、Bel-7402、MHCC97、HepG2、Hep3B、Huh-7、MS751和正常肝细胞LO2细胞及转染0,25,50,100 μmol/L miR-486-5p类似物(miR-486-5p mimic) 24 h后LO2、MHCC97、HepG2和Huh-7细胞的miR-486-5p含量;CCK-8法检测转染miR-486-5p类似物2,12,24,48,72 h的LO2、MHCC97、HepG2和Huh-7细胞的增殖率;Western Blot检测CyclinD1、Cyclin E1、CDK4、CDK6、p-AMPK、AMPK和Sift1蛋白表达量;qPCR检测转染CCND1、CCNE、CDK4、CDK6 mRNA表达量.结果:HCC患者癌组织miR-486-5p含量明显低于癌旁组织,血浆中miR-486-5p含量明显低于正常人;HCC患者癌组织和血浆中miR-486-5p含量与血浆AFP含量呈显著负相关性,相关系数分别为-0.1554和-0.2228(P＜0.001);除Huh-7细胞外,人HCC癌细胞系SMMC-7721、Bel-7402、MHCC97、HepG2、Hep3B和MS751细胞中miR-486-5p含量明显低于LO2细胞(P＜0.05);相比于阴性对照类似物(NC)处理组,0至100 μmol/L的miR-486-5p类似物处理LO2和Huh-7细胞24 h组的miR-486-5p水平均没有明显的变化(P＞0.05),50μmol/L和100 μmol/L的miR-486-5p类似物处理24 h的MHCC97和HepG2细胞,相比于NC处理组,其miR-486-5p水平均显著增加(P＜0.05);miR-486-5p类似物处理LO2组的增殖率相比于NC和Control组无明显的变化.相比于NC和Control组,miR-486-5p类似物处理HepG2和MHCC97细胞24 h的增殖率相比于2h的显著降低,分别为(77.70±7.53)％和(86.61±1.08)％.且具有时间依赖性.而miR-486-5p类似物处理Huh-7细胞48 h组增殖率为(84.79±2.33)％,明显低于2h组;miR-486-Sp类似物转染HepG2和MHCC97细胞24 h的Cyclin D1、CyclinE、CDK6蛋白与mRNA表达量明显低于NC组(P＜0.05),HepG2的miR-486-5p类似物处理CDK4相比于NC组无明显变化(P＞0.05),MHCC97的miR-486-5p类似物组CDK4相比于NC组明显降低(P＜0.05).p-AMPK与Sirt1蛋白水平均明显低于NC组.结论:过表达miR-486-5p通过下调p-AMPK/Sirt1通路阻断HCC肿瘤细胞细胞周期,并抑制癌细胞增殖.

目的 溶瘤腺病毒成为近年研究肿瘤靶向治疗的热点之一,目前鲜见溶瘤腺病毒SG600-IL24应用于裸鼠移植瘤的报道.本研究利用携带人黑色素瘤分化相关基因7/白介素24(melanoma differentiation associated gene-7/interleukin-24,MDA-7/IL-24)基因的溶瘤腺病毒SG600-IL24与干扰素联合治疗肝癌裸鼠移植瘤,旨在探讨溶瘤腺病毒基因和新辅助联合治疗肝癌的作用机制.方法 构建携带人MDA-7/IL-24基因的溶瘤腺病毒SG600-IL24,单独使用该病毒或干扰素以及两者联合治疗裸鼠肝癌模型,观察其抗肿瘤效应.裸鼠随机分为对照组、新辅助治疗组(IFN组)、基因治疗组(SG600-IL24组)和基因治疗+新辅助治疗组(SG600-IL24+ INF组).采用免疫组化方法检测各组肿瘤组织中基质金属蛋白酶-2(matrix metalloproteinase-2,MMP-2)与血管内皮生长因子(vascular endothelial growth factor,VEGF)表达情况.结果 成功构建携带人MDA-7/IL-24基因的溶瘤腺病毒SG600-IL24载体,溶瘤腺病毒SG600-IL24介导的外源基因MDA-7/IL-24在SG600-IL24组和SG600-IL24+ INF组裸鼠体内表达明显增高;对照组、IFN组、SG600-IL24组裸鼠平均生存时间分别为(35.2±1.53)、(67.6±2.10)和(59.2±1.16)d,SG600-IL24+ IFN联合治疗组有3只裸鼠生存时间超过120 d,达到长期生存,与其他3组比较生存期明显延长,F=68.20,P=0.000 1;SG600-IL24+IFN组裸鼠肝癌体积也明显小于SG600-IL24组(P=0.000 9)或IFN组(P=0.000 4);SG600-IL24联合干扰素能明显降低MMP-2[表达率为(2.0±1.9)％]和VEGF表达[MMP-2表达率为(0.8±1.1)％],作用优于SG600-IL24组、IFN组及对照组,差异有统计学意义,均P＜0.001.结论 溶瘤腺病毒SG600-IL24联合干扰素对裸鼠人肝癌种植模型有很好的协同抗肿瘤作用,其作用机制可能与抗肿瘤侵袭、转移有关.

Cholangiocarcinoma (CCA) is a cancer type with high postoperative relapse rates and poor long-term survival largely due to tumor invasion, distant metastasis, and multidrug resistance. Deregulated microRNAs (miRNAs) are implicated in several cancer types including CCA. The specific roles of the miRNA let-7c in cholangiocarcinoma are not known and need to be further elucidated. In our translational study we show that microRNA let-7c expression was significantly downregulated in human cholangiocarcinoma tissues when compared to adjacent tissues of the same patient. Let-7c inhibited the tumorigenic properties of cholangiocarcinoma cells including their self-renewal capacity and sphere formation in vitro and subcutaneous cancer cell growth in vivo. Ectopic let-7c overexpression suppressed migration and invasion capacities of cholangiocarcinoma cell lines in vitro, however, promoted distant invasiveness in vivo. Furthermore, we found that let-7c regulated the aforementioned malignant biological properties, at least in part, through regulation of EZH2 protein expression and through the DVL3/β-catenin axis. The miRNA let-7c thus plays an important dual role in regulating tumorigenic and metastatic abilities of human cholangiocarcinoma through mechanisms involving EZH2 protein and the DVL3/β-catenin axis.

OBJECTIVE:The aim of this study was to evaluate the safety, feasibility, and oncologic outcomes of 3-dimensional total laparoscopic pancreaticoduodenectomy (3D-TLPD).METHODS:Data of all patients who underwent 3D-TLPD (n = 202) or open pancreaticoduodenectomy (OPD) (n = 213) at a single institution between October 2014 and December 2016 were reviewed. We evaluated the safety, feasibility, and oncologic outcomes of 3D-TLPD compared with OPD.RESULTS:The mean operative times in the 3D-TLPD and OPD groups were comparable (P = 0.322). The estimated blood loss and perioperative transfusion rate were similar in both groups, as were the morbidity and mortality rates (P > 0.050). The mean number of analgesic administered and the mean length of hospital and intensive care unit stay were lower (P < 0.001) and shorter (P < 0.001, P = 0.009) in the 3D-TLPD group than in the OPD group, respectively. The surgical resection margins and the number of lymph nodes harvested did not differ between the 2 groups; however, a significant difference was observed in pathological results.CONCLUSIONS:Three-dimensional TLPD had the typical advantages of minimally invasive abdominal procedures, such as shorter hospital stays. It is technically safe and feasible and has comparable operative times and similar oncologic outcomes to those of OPD.

目的 探讨ALDH抑制剂DEAB致肝癌细胞凋亡及增强其对ADM化疗敏感性的具体机制.方法 通过DEAB干预肝癌细胞系HepG2和正常肝细胞系L02以及与经ADM处理的HepG2和L02细胞.通过流式细胞仪检测各组细胞中活性氧(ROS)的表达变化及Annexin-V和PI双染后细胞凋亡量的变化.通过Realtime PCR及western blot方法检测各组细胞中Bcl-2、Bax、ALDH1以及MDR1基因和蛋白的变化.结果 DEAB干预HepG2细胞后,致使其ROS表达增强、凋亡量增加,ALDH1基因和蛋白表达无明显变化,Bcl-2、MDR1基因和蛋白表达下调,Bax基因和蛋白表达上调.正常肝细胞系L02无以上变化.结论 抑制ALDH可致肝癌细胞系HepG2凋亡,并增强其对ADM的化疗敏感性.而对正常肝细胞系L02无此作用.

BACKGROUND:The treatment of borderline resectable pancreatic head cancer (BRPHC) is still controversial and challenging. The artery-first approaches are described to be the important options for the early determination. Whether these approaches can achieve an increase R0 rate, better bleeding control and increasing long-term survival for BRPHC are still controversial. We compared a previously reported technique, a modified artery-first approach (MAFA), with conventional techniques for the surgical treatment of BRPHC.METHODS:A total of 117 patients with BRPHC undergone pancreaticoduodenectomy (PD) from January 2013 to June 2015 were included. They were divided into an MAFA group (n=78) and a conventional-technique group (n=39). Background characteristics, operative data and complications were compared between the two groups.RESULTS:Mean operation time was significantly shorter in the MAFA group than that in the conventional-technique group (313 vs 384 min; P=0.014); mean volume of intraoperative blood loss was significantly lower in the MAFA group than that in the conventional-technique group (534 vs 756 mL; P=0.043); and mean rate of venous resection was significantly higher in the conventional-technique group than that in the MAFA group (61.5% vs 35.9%; P=0.014). Pathologic data, early mortality and morbidity were not different significantly between the two groups.CONCLUSIONS:MAFA is safe, simple, less time-consuming, less intraoperative blood loss and less venous resection, and therefore, may become a standard surgical approach to PD for BRPHC with the superior mesenteric vein-portal vein involvement but without superior mesenteric artery invasion.

Objective To investigate the effect of oncolytic adenovirus vector SG600-IL24 expressing human melanoma differentiation associated gene-7/interleukin-24 (MDA-7/IL-24) on hepatocellular carcinoma cell lines with different metastatic potentials (HepG2,Hep3B,SMMC7721 and normal liver cell line L02).Methods The oncolytic adenovirus SG600-II24 carrying MDA-7/IL-24 gene was transfected into hepatocellular carcinoma cell lines and normal liver cell line.The mRNA and protein expression of MDA-7/IL-24 in HepG2,Hep3B,SMMC7721 and L02 cells was confirmed by reverse transcriptase-polymerase chain reaction (RT-PCR),enzyme linked immunosorbent assay (ELISA) and Western blotting respectively.Methyl thiazol tetrazolium (MTT) assay and flow cytometry were used to study tumor cell proliferation and cell cycle in vitro.Hoechst 33258 and flow cytometry were studied to indicate the apoptosis effects.Results It was confirmed by RT-PCR,ELISA and Western blotting that the exogenous MDA-7/IL-24 gene was highly expressed in HepG2,Hep3B,SMMC7721 and L02 cells.MTT and apoptosis detection indicated that MDA-7/IL-24 could induce the growth suppression [inhibition rate:(75.0 ±2.5)％,(85.0 ±2.0)％,and (86.0±3.5)％ respectively.for HepG2:F=5.860;for Hep3B:F=7.210;for SMMC7721:F=7.980,P=0.000],promote apoptosis [apoptosis rate:(56.5 ± 4.0) ％,(34.4 ± 2.0) ％ and (43.3 ± 2.5) ％ respectively.For HepG2:F =203.400;for Hep3B:F =313.200;for SMMC7721:F=130.500,P =0.000],and block cancer cells in G2/Mphase [the blocking rate:(35.4±4.2)％,(47.3±6.2)％ and (40.5±5.0)％] but not L02 cells.Conclusion Oncolytic adenovirus vector SG600-IL24 can selectively induce growth suppression and promote apoptosis in hepatocellular carcinoma lines in vitro but not in L02 cells.

目的 探讨罕见病胰高血糖素瘤的诊断治疗方法.方法 回顾性分析本院收治的2例及国内文献报道共42例胰高血糖素瘤的诊治资料.结果 所有患者在病程中的不同阶段曾被误诊.国内文献报道的42例患者中92.9%患者有典型的胰高血糖素瘤综合征,33例行手术治疗,病理诊断恶性者26例;所有手术患者术后皮疹均未见新发;1例术后局部及肝脏复发行减瘤手术后仍带瘤生存.结论 胰高血糖素瘤临床罕见,极易误诊.根据典型的胰高血糖素瘤综合征及实验室检查血浆胰高血糖素水平即可提出定性诊断,辅以影像学检查可提出定位诊断.根治性或姑息性手术切除是其有效的治疗方法.

The technique of minimally invasive pancreatic surgeries has evolved rapidly, including minimally invasive pancreaticoduodenectomy (MIPD). However, controversy on safety and feasibility remains when comparing the MIPD with the open pancreaticoduodenectomy (OPD); therefore, we aimed to compare MIPD and OPD with a systemic review and meta-analysis.