Figure S1 - PDF file 1773K, miR-200a expression levels is unchanged in human gastric cancer.(A) The expression levels of miR-200a was detected in 36 gastric cancer patients by qRT-PCR. Data are shown as fold change of gastric cancer relative to adjacent normal tissues. (B) Relative expression of miR-200a in 7 cell lines derived from gastric cancer and one nonmalignant gastric cell line (GES-1) was determined by qRT-PCR. Data are presented as mean SD from at least three separate experiments

Agmatinase (AGMAT) is an enzyme that hydrolyzes agmatine to putrescine and urea. In this study, we explored the functions of AGMAT in colorectal cancer (CRC). By performing gain-of-function and loss-of-function experiments, we investigated the roles of AGMAT in proliferation, cell cycle progression, and apoptosis of CRC cells. We also established a colitis-associated colorectal cancer model by challenging mice with azoxymethane (AOM) and dextran sodium sulfate (DSS), and we subsequently silenced AGMAT expression in mice by adeno-associated virus 9 (AAV9)-mediated delivery of short hairpin RNA (shRNA). In vitro experiments showed that overexpression of AGMAT accelerated the proliferation and inhibited the apoptosis of CRC cells, and AGMAT knockdown exhibited the opposite effects. Interestingly, the oncogenic transcription factor, c-Myc, could bind to the AGMAT promoter and transcriptionally increase AGMAT expression in CRC cells. Additionally, c-Myc and AGMAT were upregulated in the colon of AOM/DSS-treated mice, and AGMAT silencing significantly mitigated colitis in AOM/DSS-treated mice, as evidenced by the increased colon length, attenuated crypt damage, and reduced levels of inflammatory indicators (myeloperoxidase, interleukin-6, tumor necrosis factor-α, inducible nitric oxide synthase, and phosphorylated p65) in colon tissues. Notably, AGMAT silencing decreased both the number and size of tumors, reduced expression of proliferating cell nuclear antigen, and inhibited phosphorylation of protein kinase B (AKT) and extracellular signal-regulated kinase in the colon of AOM/DSS-treated mice. Overall, we determined that AGMAT facilitates tumor progression in CRC. Our findings will be helpful in the search for potential therapeutic targets for CRC.

目的 探究羟基氯喹对人结肠癌SW480细胞增殖及凋亡的影响,并分析可能的机制.方法 体外培养人结肠癌细胞系SW480细胞,随机分为空白组和低、中、高剂量实验组.空白组正常培养,低、中、高剂量实验组分别用加入含羟基氯喹终浓度为20,40,80μg·mL-1的细胞培养液.用噻唑蓝(MTT)法检测细胞增殖活性;用平板克隆实验检测细胞克隆形成率;用AnnexinV-FITC/PI双染法检测细胞凋亡情况;用蛋白质印迹法检测Toll样受体4(TLR4)、核转录因子kappa B p65(NF-κB p65)、半胱氨酸的天冬氨酸蛋白酶-3(Caspase-3)蛋白表达水平.结果 空白组和低、中、高剂量实验组SW480细胞存活率分别为100％,(83.15±12.45)％,(62.01±9.32)％和(43.25±6.45)％;这4组的克隆形成率分别为(85.16±12.77)％,(65.28±9.78)％,(51.06±7.07)％和(37.11±5.57)％;这4组的凋亡率分别为(9.12±1.37)％,(16.72±2.51)％,(25.89±3.89)％和(39.75±5.97)％;这4组的TLR4蛋白相对表达水平分别为1.15±0.18,0.90±0.13,0.65±0.09和0.29±0.05;这4组的NF-κB p65蛋白相对表达水平分别为1.27±0.20,0.96±0.15,0.69±0.11和0.47±0.08;这4组的Caspase-3蛋白相对表达水平分别为0.43±0.07,0.66±0.09,1.01±0.16和1.38±0.21.上述指标,低、中、高剂量组与空白组比较,差异均有统计学意义(均P<0.05).结论 羟基氯喹可能通过抑制TLR4/NF-κB通路活化抑制人结肠癌SW480细胞增殖并诱导细胞凋亡.

Lymph node involvement is one of the most important prognostic factors for colorectal cancer. Para-aortic lymph node metastasis (PALNM) is an uncommon mode of metastasis of colorectal cancer. Because the incidence of colorectal cancer is lower than that of liver and lung metastases, related studies have certain limitations. Therefore, the diagnosis and treatment of PALNM in colorectal cancer are unclear. Para-aortic lymph node dissection (PALND) can provide a pathological diagnosis and can also improve the prognosis of some selected patients. Although aggressive surgical resection of colorectal cancer with liver and lung metastases is indicated, the optimal surgical management strategy for colorectal cancer with PALNM remains unclear. To determine whether PALND should be performed, the relationship between its safety, effectiveness, recurrence rate, and survival benefit needs to be assessed. This study reviews the research and treatment progress of PALNM in colorectal cancer in recent years.

Ferroptosis, a newly iron-dependent form of cell death, is often accompanied by the damage of membrane lipid peroxide. Recently, the ferroptosis inducer erastin has been reported to exhibit potential anti-cancer activities. The aim of this study was to investigate the effects of SRSF9 on the sensitivity of colorectal cancer (CRC) to erastin and explore the underlying molecular mechanism. Short hairpin RNAs (shRNAs) or SRSF9 overexpression vector (SRSF9-OE) was transfected into erastin-induced human CRC cells to inhibit or overexpress SRSF9. Results showed that SRSF9 inhibition promoted the cell death induced by erastin, conversely, SRSF9 overexpression augmented the resistance to erastin-induced death in human CRC cells. SRSF9 decreased lipid peroxide damage which was a key event during erastin-induced ferroptosis in human CRC cells. Furthermore, we found that SRSF9 inhibition increased erastin-induced ferroptosis by downregulating GPX4 level. In an In vivo study, SRSF9 shRNA or SRSF9-OE stably transfected human CRC cells were subcutaneously injected into the right flank of nude mice. SRSF9 overexpression partly abolished the tumor growth inhibition and ferroptosis induced by erastin. Our data indicated SRSF9's regulation of GPX4 as an essential mechanism driving CRC tumorigenesis and resistance of erastin-induced ferroptosis. This molecular mechanism may provide a novel method for improving the sensitivity of CRC to erastin.

To discuss recurrence patterns and their significance in colorectal cancer. Preexisting medical hypotheses and the clinical phenomena of recurrence in colorectal cancer were evaluated and integrated. Colorectal cancer recurrence/metastasis consists of two types: recurrence from the activation of dormant cancer cells and recurrence from postoperative residual cancer cells. These two recurrences have their own unique mechanisms, biological behaviors, responses to therapy, and prognoses. For type 1 recurrences, surgical resection should be considered. Type 2 recurrences should be managed systematically in addition to surgical resection. The two types of colorectal cancer recurrence should be evaluated and managed separately.

Ferroptosis, a newly discovered form of programmed cell death characterized by lipid peroxidation, crafts a new perspective on cancer treatment. Serine and arginine rich splicing factor 9 (SFRS9) is frequently described as a proto-oncogene in cervical and bladder cancer. However, the role of SFRS9 in colorectal cancer (CRC) and whether SFRS9 exerts its function associated with ferroptosis is largely unknown. Herein, we found that the expression of SFRS9 mRNA and protein in the CRC tissues was obviously higher than that in the paracancerous tissues. Function assays revealed that SFRS9 overexpression (SFRS9-OE) significantly promoted cell viability, cell cycle progression and colony formation of CRC cells. While SFRS9 knockdown by shRNAs transfection inhibited these progressions. Furthermore, cell death and lipid peroxidation induced by ferroptosis inducers erastin and sorafenib were suppressed by SFRS9-OE. Bioinformatics analysis indicated that SFRS9 can bind to peroxidase 4 (GPX4) mRNA which is a central regulator of ferroptosis. Western blot showed that GPX4 protein expression was clearly elevated upon SFRS9-OE, while it was decreased in SFRS9-inhibited CRC cells. RNA immunoprecipitation experiment was carried out in HCT116 cells to confirm the binding of SFRS9 and GPX4 mRNA specifically. SiGPX4 transfection reversed the inhibitory effects of SFRS9-OE on the erastin and sorafenib-induced ferroptosis. Consistent with our in vitro observations, SFRS9 promoted the growth of tumors while SFRS9 knockdown significantly inhibited tumor growth in nude mice. In conclusion, SFRS9 represents an obstructive factor to ferroptosis by upregulating GPX4 protein expression, and knocking down SFRS9 might be an effective treatment for CRC.

Purpose: The inflammatory response plays a crucial role in the occurrence and development of colon cancer. In this study, we aimed to explore a novel prognostic model for patients with colon cancer (COAD) based on inflammatory response-related genes. Methods: Inflammatory response-related genes were obtained from Molecular Signatures database. Univariate and multivariate Cox regression analyses were used for model construction based on TCGA dataset. GSE39582 dataset and qRT-PCR dataset were used for validation. Gene set variation analysis and gene set enrichment analysis were performed to explore the potential regulatory pathways. The immune cell infiltration level was analyzed via CIBERSORT. Immunohistochemistry analysis and experiments were used to explore the function of genes in model. Results: In this study, a novel prognostic signature was identified using stepwise Cox proportional hazards regression analysis based on TCGA dataset. The results were subsequently validated in 562 patients from GSE39582 and a qRT-PCR data set from 70 tumor samples. Functional analysis indicated that the tumor microenvironment and immune cell infiltrate were different between high-and low-risk groups. Additionally, IHC results showed that the protein levels of prognostic genes were significantly different between COAD tissues and adjacent non-tumorous tissues, and prognostic genes could regulate the malignant phenotype of COAD cells. Conclusion: Overall, the inflammation-related gene signature can be used for prognostic prediction in patients with COAD.

Cancer stem cells play crucial roles in the development of colon cancer (COAD). This study tried to explore new markers for predicting the prognosis of colon cancer based on stem cell-related genes. In our study, 424 COAD samples from TCGA were divided into three subtypes based on 412 stem cell-related genes; there were significant differences in prognosis, clinical characteristics, and immune scores between these subtypes. 694 genes were screened between subgroups. Subsequently a six-gene signature (DYDC2, MS4A15, MAGEA1, WNT7A, APOD, and SERPINE1) was established. This model had strong robustness and stable predictive performance in cohorts of different platforms. Taken together, the six-gene signature constructed in this study could be used as a novel prognostic marker for COAD patients.

The karyopherin α2 subunit gene (KPNA2), an oncogene, is involved in metabolic reprogramming in cancer. This study aimed to explore the function of KPNα2 in the growth and glycolysis in colon cancer (CC) cells. Genes from the Oncomine database that were differentially expressed in multiple CC types were screened. Bioinformatics analysis suggested that KPNA2 was highly expressed in CC, and consequently, high expression of KPNA2 was detected in the CC cell lines. Down-regulation of KPNA2 reduced viability and DNA-replication ability, and increased apoptosis of HCT116 and LoVo cells. It also reduced glucose consumption, extracellular acidification rate, and the ATP production in cells. Centromere protein A (CENPA) was confirmed as an upstream transcription activator of KPNA2. There was significant H3K27ac modification in the promoter region of KPNA2. CENPA primarily recruited histone acetyltransferase general control of amino acid synthesis (GCN)-5 to the promoter region of KPNA2 to induce transcription activation. Overexpression of either CENPA or GCN-5 blocked the role of short hairpin KPNα2 and restored growth and glycolysis in CC cells. To conclude, the findings from this study suggest that CENPA recruits GCN-5 to the promoter region of KPNA2 to induce KPNα2 activation, which strengthens growth and glycolysis in, and augments the development of, CC.

Abstract Introduction: Vaginal agenesis is a congenital disorder, which can be managed by nonsurgical dilation or surgical reconstruction of the vagina. The sigmoid vaginoplasty procedure is a popular approach, which pulls down part of the sigmoid colon to form a neovagina. One complication of this procedure is introital stenosis. Patient concerns: A 55-year-old woman presented to the outpatient general surgery department with severe, persistent abdominal pain. The patient was diagnosed with congenital absence of uterus and vagina, and a sigmoid vaginoplasty was performed 34 years ago. Diagnosis: A pelvic MRI and an abdominal enhanced CT scan were performed, finding that the uterus was absent, and the os of the vagina was closed, forming a closed loop full of fluid. Introital atresia and closed loop of neovaginal colon conduit were diagnosed. Interventions: Based on our conclusions and the patient's consent we surgically removed the neovagina. Outcomes: After surgery, the abdominal pain was relieved, and the patient reported full recovery during a 6-month follow-up appointment. Conclusion: Introital stenosis is one of the long-term complications of sigmoid vaginoplasty procedure. Introital stenosis, leading to introital atresia, is rare but may occur. Surgical removal of neovagina can relieve the pain in patients who do not have the demand of sexual intercourse.

目的:分析慢性便秘患者的腹部CT影像学表现,探讨CT检查对慢性便秘的诊断价值.方法:收集中国医科大学附属盛京医院2012年2月-2017年7月因慢性便秘为主诉患者的CT影像学资料,共68例,分析胃肠道的一般影像学特征,包括肠道的形态、直径、粪便分布等特点.结果:CT表现提示肠壁形态僵硬23例(33.82％),结肠袋消失30例(44.12％),结肠最大直径大于6 cm即符合巨结肠诊断标准17例(6.2～15.3 cm,平均9.09 cm),可见肠道移行段7例(10.29％),CT可见肠腔内高密度粪块或大量低密度内容物(油性通便剂)63例(92.65％),相对冗长且大量粪便分布于升结肠段4例(5.88％)、横结肠30例(44.12％)、降结肠2例(2.94％)、乙状结肠20例(29.41％)、结肠肝曲和脾曲12例(17.65％).结论:CT可作为便秘患者的常规检查,有一定的诊断价值.

目的 探索案例教学法(case-based learning,CBL)联合多学科诊疗模式(multi-disciplinary treatment,MDT)的教学模式在结直肠外科住院医师规范化培训中的应用效果.方法 将2017年6月—2018年12月在中国医科大学附属盛京医院结直肠外科进行住院医师规范化培训的58名住院医师为研究对象,随机分为试验组及对照组,每组各29人,试验组采用CBL联合MDT教学法,对照组采用传统教学法,通过客观成绩测定、学生主观评价教学满意度来评估不同教学方法的教学效果.结果 试验组的基础理论成绩和病例分析成绩均高于对照组,差异有统计学意义(P<0.05),临床操作成绩与传统教学组比较差异无统计学意义(P>0.05);教学满意度调查,试验组高于对照组,具有统计学意义(P<0.05).结论 联合MDT的CBL教学模式凭借其整合性强及理论结合实践等优势,能够有效提高肛肠科住院医师规范化培训的效果.

INTRODUCTION:Slow transit constipation is a major cause of chronic constipation. During pregnancy, changes in hormone levels and the physical effects of an enlarged uterus could cause new onset slow transit constipation or aggravate a pre-existing constipation. The management of slow transit constipation-induced ileus during pregnancy is a medical dilemma. PATIENT CONCERNS:A 28-year-old pregnant woman presented to the emergency department with a 7-day history of worsening bloating and abdominal colic. The patient was in her third trimester (27 weeks). She had a 5-year history of constipation which had worsened with her pregnancy, and neither flatus nor stool could be passed. DIAGNOSIS:Based on the constipation history and computed tomography, a slow transit constipation-induced ileus was confirmed. INTERVENTIONS:As medications for the management of constipation and endoscopic efforts to remove the blockage were ineffective and the patient's symptoms worsened, Cesarean section and colectomy with ileorectal anastomosis were performed. OUTCOMES:After the procedure, the patient recovered and defecated well. At the 6-month follow-up, the patient reported that she defecated two to three times per day without difficulty. CONCLUSION:Pregnancy can worsen pre-existing constipation and cause ileus. In cases where drug treatment is unsuccessful, colectomy, and ileorectal anastomosis may be necessary.

Objective To explore the clinical value of neutrophil/lymphocyte ratio (NLR) in staging of stage Ⅱ/Ⅲ middle and low rectal cancer. Methods The clinical and pathological data of 111 patients with middle and low rectal cancer who underwent surgical treatment were retrospectively analyzed. The NLR values along with various clinical factors and pathological parameters were also statistically analyzed, and then the relationship between preoperative NLR and various clinical factors was determined. All patients were divided into high and low groups based on NLR, with the cut-off value of 1.99. Then, the clinical, pathological parameters and disease-free survival of the two groups were analyzed. Results The maximum diameter of the tumor and the NLR value were positively correlated (P ＜ 0.05). The patients with high and low NLR had significant differences in age, stage Ⅱ and Ⅲ, N stage, pathological type, maximum diameter of the tumor, and disease-free survival (P ＜ 0.05). Conclusion Preoperative NLR values may be used as predictive values for the pathological parameters of stage Ⅱ and Ⅲ middle and low rectal cancer, and NLR may be an indicator of its prognostic assessment.

目的 探讨淋巴示踪在腹腔镜结直肠癌根治手术中的应用效果及对微小淋巴结检出的影响.方法 回顾性分析80例腹腔镜结直肠癌根治术患者资料,根据使用示踪剂情况分为对照组(40例)和观察组(40例),观察组术前使用淋巴示踪剂,对照组术前则不使用,观察对比两组的手术基本情况、淋巴结情况、1年生存率与复发率以及肿瘤标志物情况.结果 观察组手术基本情况优于对照组,差异有显著性(P<0.05);淋巴结清扫数、有转移的淋巴结清扫数和微小淋巴结检出数均较对照组多(t=-15.539、-10.670、-15.888,P<0.001);两组1年生存率与复发率比较,差异无显著性(P>0.05);治疗后观察组癌胚抗原、细胞角蛋白19片段抗原和糖链抗原125水平低于对照组(t=14.503、9.413、12.381,P<0.001).结论 淋巴示踪在腹腔镜结直肠癌根治手术中的应用效果较好,可明显提高微小淋巴结检出率.

结直肠癌是世界范围内常见的恶性肿瘤.随着手术技术的进步以及新的治疗药物的出现,结直肠癌的5年生存率逐渐提高,但结直肠癌仍是导致肿瘤相关性死亡的重要原因之一.结直肠癌具有高度异质性,其生物学行为在不同个体间或同一个体的不同病灶间具有广泛的遗传学和表观遗传学差异,可进一步导致患者预后以及对治疗反应的不同.在分子层面对结直肠癌进行分型,了解每种亚型的分子改变特征及其相应的肿瘤生物学行为特征,对结直肠癌的治疗具有重要的指导意义.

目的:探讨术前CT的骨盆测量解剖参数及病理参数对腹腔镜中低位直肠癌手术难度预测价值及相应预测评分系统建立.方法:纳入2014年10月至2018年4月于中国医科大学附属盛京医院结直肠肛门外科行腹腔镜直肠癌根治术的80例患者,通过对术前CT骨盆测量得到的解剖参数及病理参数进行单因素分析得到可能影响手术难度的因素,再将筛选出的危险因素进行logistic多因素分析,根据logistic回归分析特点对危险因素赋值,建立评分系统,再利用受试者工作曲线(receiver operating characteristic,ROC)评价该评分系统效能.结果:肿瘤位置、耻骨联合高度比中骨盆前后直径、耻骨联合高度比中骨盆棘突直径为手术操作难度的影响因素.通过以上述因素构建的评分系统,得分越高,困难程度越大.结论:更低的肿瘤位置及更深窄的骨盆可能会导致手术难度增加.相应的评分系统有助于术前对患者进行评估,选择合适的术式.

Although increasing evidence have confirmed that carbon monoxide release molecule-2(CORM-2) plays an active role in the treatment of inflammation and tumors, poor aqueous solubility and short CO-release duration restrict its extensive application. Our previous work synthesized styrene-maleic acid copolymer-encapsulated CORM-2 (SMA/CORM-2) to overcome above-mentioned deficiencies and demonstrated satisfactory effects in colitis. This study is to investigate the function of SMA/CORM-2 on colorectal cancer proliferation and metastasis. CCK-8 experiment is used to clarify the half maximal inhibitory concentration (IC50) of SMA/CORM-2 and to detect cell proliferation. Transwell assay coated with or without matrigel was to detect cell invasion and migration. Western blot was used to detect β-catenin, AKT, p-AKT, VEGF, MMP-2 and MMP-9 proteins. At last, nude mice xenograft was used to further investigate the anti-tumor effect of SMA/CORM-2 in vivo. After SW480 and C26 cells were treated with 0.5 mg/ml SMA/CORM-2, CRC cells proliferation, migration and invasion were inhibited. In vivo, SMA/CORM-2 treatment remarkably suppressed tumor growth and lung metastasis in nude mice. Furthermore, the expression of β-catenin, p-AKT, VEGF, MMP-2 and MMP-9 proteins could be down-regulated after SMA/CORM-2 treatment. SMA/CORM-2 exerted both in vitro and in vivo anti-proliferation and anti-metastatic effects, which may yield a novel therapeutic strategy for CRC.

目的 通过Ualcan数据库分析WASH2P在结直肠癌中的表达及意义.方法 利用UALCAN数据库分析结肠癌中WASH2P的表达水平,并分析其与肿瘤分期、种族、性别、体重、病理亚型之间的关系;对WASH2P进行生存分析,并利用GEPIA数据库进行验证;对结肠癌中WASH2P与MSH5和PMS2的协同表达情况进行分析.结果 WASH2P在结肠癌组织中的表达高于正常组织,WASH2P高表达的结肠癌患者预后差,WASH2P与MSH5、PMS2的表达存在明显的相关性.结论 WASH2P可能是治疗结肠癌的潜在靶点.