Studies have shown that UHRF1 can affect the expression of target genes by regulating DNA methylation, which may be related to the mechanism of cancer cell death, especially ferroptosis. In this study, HuH7 and HepG2 hepatoma cell lines were used. Intracellular GSH/GSSG and NADP+/NADPH levels were assessed to analyze REDOX status, and lipid peroxidation was assessed using lipid peroxidation level assays. The regulation of GSTZ1 by UHRF1 and its modification through DNA methylation were explored. UHRF1 silencing significantly inhibited the growth and reproduction of liver cancer cells, led to iron death in HuH7 and HepG2 cells, and caused REDOX imbalance and lipid peroxidation. Further analysis showed that UHRF1 regulates iron death in liver cancer cells by mediating the expression of GSTZ1. UHRF1 can down-regulate the expression of GSTZ1 through DNA methylation modification. The study revealed that UHRF1 protein inhibits the expression of GSTZ1 by regulating DNA methylation, thereby affecting the iron death of liver cancer cells.

Introduction: Colorectal cancer (CRC) is hackneyed cancer and a major lethiferous cancer. Circular RNAs (CircRNAs) have been discovered to own important roles in controlling CRC progression. CircPSMC3 is known to exhibit lower expression in diversified cancers. However, the regulatory function of CircPSMC3 in CRC keeps unclear. Methods: The expression of CircPSMC3 and miR-31-5p was confirmed through RT-qPCR. The cell proliferation was measured through CCK-8 and EdU assays. The protein expression of genes was examined through a western blot. The cell invasion and migration were tested through Transwell and wound healing assays. The binding ability between CircPSMC3 and miR-31-5p was confirmed through the luciferase reporter assay. Results: CircPSMC3 exhibited lower expression in CRC tissues and cell lines. Additionally, CircPSMC3 was revealed to suppress cell proliferation in CRC. Moreover, through Transwell and wound healing assays, CircPSMC3 was discovered to repress CRC cell invasion and migration. In CRC tissues, miR-31-5p expression was up-regulated and negatively correlated with CircPSMC3 expression. Further mechanism exploration experiments disclosed that CircPSMC3 is bound with miR-31-5p to modulate the YAP/β-catenin axis in CRC. At last, through rescue assays, CircPSMC3 inhibited cell proliferation, invasion and migration through sponging miR-31-5p in CRC. Conclusion: Our work was the first time to probe the potential regulatory effects of CircPSMC3 in CRC, and these above results uncovered that CircPSMC3 inhibited CRC cell growth and migration through regulating miR-31-5p/YAP/β-catenin. This discovery hinted that CircPSMC3 may serve as a useful therapeutic candidate for CRC.

The nuclear receptor 4A1 (NR4A1) is widely involved in the regulation of cell survival and is related to ischemic injury in several organs. This research examined the emerging role and mechanism of NR4A1 in hepatocyte ischemia-reperfusion injury (IRI). BRL-3A cells were subjected to hypoxia-reperfusion (H/R) to simulate an IRI model in vitro. The expression of NR4A1 and liver kinase B1 (LKB1)/AMP-activated protein kinase (AMPK) pathway-related proteins (LKB1, AMPK, and ACC) was detected by western blotting or RT-qPCR under H/R condition after NR4A1 overexpression or silencing. Then, radicicol, an inhibitor of LKB1 pathway, was used to determine the role of NR4A1 in hepatocyte H/R injury by regulating LKB1. Under the help of CCK-8 assay, cell viability was assessed. The levels of ROS, MDA, and SOD were determined with corresponding kits to evaluate oxidative stress. Additionally, RT-qPCR was employed to analyze the releases of the inflammatory factors. Flow cytometry was applied to estimate the apoptosis and its related proteins, and autophagy-associated proteins were assayed by western blotting. Results indicated that NR4A1 was highly expressed, while proteins in LKB1/AMPK signaling was downregulated in BRL-3A cells exposed to H/R. The activation of LKB1/AMPK pathway could be negatively regulated by NR4A1. Moreover, NR4A1 depletion conspicuously promoted cell viability, inhibited oxidative stress as well as inflammation, and induced apoptosis and autophagy in H/R-stimulated BRL-3A cells, which were reversed after radicicol intervention. Collectively, NR4A1/LKB1/AMPK axis is a new protective pathway involved in hepatocyte IRI, shedding new insights into the improvement of hepatocyte IRI.

Kinesin family member 2A (KIF2A) serves a vital role in the development of hepatocellular carcinoma (HCC); however, the biological effect of KIF2A on the malignant progression of HCC remains unclear. Therefore, the present study was conducted to systematically determine the biological role of KIF2A in HCC and to better understand the molecular mechanism. The differences of KIF2A expression in HHL-5 normal human hepatocytes and the HCC cell lines Li-7, Huh7 and MHCC97 were assessed by reverse transcription-quantitative PCR and western blotting analysis. Moreover, viability, proliferation, migration and invasion of HCC cells were assessed by performing CCK-8, 5-ethynyl-2'-deoxyuridine staining, wound healing and Transwell assays. Additionally, the tube formation assay was performed to evaluate angiogenesis of HUVECs incubated with the conditioned media of HCC cells in vitro. The interaction between KIF2A and Notch1 was analyzed through co-immunoprecipitation assay. KIF2A was revealed to be highly expressed in HCC cells. KIF2A knockdown suppressed HCC cell proliferation, migration and invasion, and impaired in vitro angiogenesis. Furthermore, it was revealed that KIF2A interacted with Notch1 and positively regulated Notch1 expression. The suppressive effects of KIF2A knockdown on HCC cell proliferation, migration, invasion and in vitro angiogenesis were partially reversed by Notch1 overexpression. Overall, KIF2A may act as an oncogene in HCC via activation of the Notch1 signaling pathway.

Background: Circ_0026344 was reported to be associated with the metastasis of colorectal cancer (CRC). This study aimed to investigate the expression of circ_0026344 in CRC and the effect mechanisms of circ_0026344 on CRC. Methods: The expressions of circ_0026344 and miR-31 in clinical CRC tissues or CRC cell lines were analyzed by qPCR. The target of circ_0026344 was predicted and verified by CircInteractome and dual-luciferase reporter assays. The correlation between circ_0026344 and miR-31 expression was analyzed using Pearson analysis. After the CRC cells were overexpressed circ_0026344 or miR-31 or silenced circ_0026344, the viability, apoptosis, migration, and invasion of CRC cells were evaluated by CCK-8, flow cytometry, wound healing, and transwell. Also, the expressions of miR-31, Bcl-2, Bax, E-cadherin, and N-cadherin in the cells were detected by qPCR or Western blot. Results: Circ_0026344 was low-expressed in CRC tissues and cell lines. Circ_0026344 sponged miR-31 which was high-expressed in CRC tissues. The expression of circ_0026344 was negatively correlated to the expression of miR-31. The miR-31 expression could be down-regulated by circ_0026344 overexpression. Circ_0026344 overexpression inhibited the cell viability, migration, and invasion; and enhanced the apoptosis of CRC cells. Circ_0026344 overexpression decreased the expressions of Bcl-2 and N-cadherin and increased the expressions of Bax and E-cadherin in CRC cells. Circ_0026344 silencing and miR-31 overexpression had an opposite effect on CRC cells as circ_0026344 overexpression. Furthermore, miR-31 overexpression counteracted the effect of circ_0026344 overexpression. Conclusion: Circ_0026344 overexpression inhibited the migration, invasion, and enhanced apoptosis of CRC cells by sponging miR-31.

目的 探讨引起结直肠癌术后发生肠梗阻发生的危险因素.方法 回顾性分析行结直肠癌根治术的420例患者的临床资料,通过非条件Logistic回归分析可能引起术后肠梗阻的危险因素.结果 N2分期、右半结肠癌切除术、左半结肠癌切除术、结直肠肿瘤切除病史、腹腔感染、腹腔灌注化疗、术前合并肠梗阻及手术时间≥2.5 h为术后肠梗阻发生的独立危险因素(OR＞1,均P＜0.05);腹腔镜手术、术前白蛋白≥30 g/L为术后肠梗阻发生的独立保护因素(OR＜1,均P＜0.05).结论 结直肠癌术后发生肠梗阻由多种因素共同作用,积极有效的处理这些因素助于预防和减少术后肠梗阻的发生.

Background: Colorectal cancer (CRC) is considered as one of the most lethal malignancies worldwide. However, the effective therapies remain limited. Polydatin, a main effective component of the Chinese herb Polygonum cuspidatum, has multiple antitumor activities; however, whether Polydatin has anti-CRC activity is not fully understood. Materials and Methods: CRC cell proliferation and apoptosis were measured after treatment of Polydatin using Cell Counting Kit-8 assay, colony formation assay, and flow cytometer assay. The expression of miR-382 and programmed cell death ligand 1 (PD-L1) were determined in CRC cell lines by quantitative real-time polymerase chain reaction and western blot, respectively. Furthermore, dual-luciferase reporter assay was conducted to determine the target of miR-382. Moreover, loss-of-functional experiments were used to identify the effect of Polydatin on miR-382. Finally, tumor xenograft experiments were conducted to determine the effect of Polydatin in vivo. Results: As a result, Polydatin effectively inhibited cell proliferation and promoted cell apoptosis in CRC cell lines. PD-L1 was confirmed as a direct target of miR-382. Furthermore, Polydatin could suppress the expression of PD-L1 by upregulating miR-382. Moreover, Polydatin inhibits proliferation and promotes apoptosis of CRC cells by regulating miR-382 and suppressing CRC tumor growth in vivo. Conclusion: Polydatin inhibits CRC cell proliferation and promotes apoptosis by regulating miR-382/PD-L1 axis. Polydatin could be a potential compound to synthesize novel antitumor drugs.

Purpose: To investigate the effect of cytokine-induced killer cells (CIKs) immunotherapy on colorectal cancer cells and the possible mechanisms using co-culture model of CIKs and human colorectal cancer cells. Methods: CIKs were induced by INF-gamma, CD3 and recombinant human IL-2 culture system in vitro. A co-culture model of CIKs and SW480 colorectal cancer cell line was established The proliferation, invasion and apoptosis of SW480 cells were detected by CCK-8 (cell counting kit-8), transwell assay and Hoechst staining, respectively. The contents of IL-2, IL-10, insulin-like growth factor (IGF)-beta and vascular endothelial growth factor (VEGF) in the culture medium was detected by ELISA (enzyme-linked immunosorbent assay). The expression levels of AMPK/Akt/mTOR signaling-related proteins were detected by Western blot. Results: CCK-8 assay showed that the proliferative ability of co-cultured CIK+SW480 cells was remarkably lower than that of SW480 cells (p <0.05). Transwell assay showed that the number of invasive cells in co-cultured CIK+SW480 cells was less than that of SW480 cells (p<0.05). Hoechst staining showed that the apoptosis rate of co-cultured CIK+SW480 cells was higher compared with that of SW480 cells (p<0.05). Increased levels of IL-2 and IL-10 and decreased levels of IGF-beta and VEGF were found in the co-culture medium by ELISA (p<0.05). Western blot results indicated that p-AMPK and p-Akt were up regulated, whereas FoxM1 and p-mTOR were downregulated in co-cultured CIK+SW480 cells compared with SW480 cells (p <0.05). Conclusions: CIKs inhibit the proliferation and invasion of colorectal cancer cells through downregulating FoxM1 and mTOR via AMPK/Akt/mTOR pathway.

目的 探讨腹腔镜微波消融术联合三维适形放疗治疗肝癌转移患者的临床疗效.方法 选择肝癌转移患者89例的资料进行回顾性分析,按照患者的治疗方法分为对照组(n=45)和观察组(n=44),对照组患者给予三维适形放疗,观察组患者在对照组基础上使用腹腔镜微波消融术,对比两组的近期治疗效果、治疗前后的生活质量、治疗过程中的各项不良反应以及总生存时间.结果 观察组患者的治疗效果、治疗有效率和疾病控制率均优于对照组,差异有统计学意义(P＜0.05).经过治疗后,两组患者的生活质量均提升,观察组的升高幅度大于对照组(P＜0.05).两组患者的发热[37.78％(17/45)vs.45.45％ (20/44)]、疼痛[51.11％ (23/45)vs.61.36％ (27/44)]、白细胞下降[40.00％(18/45)vs.34.09％ (15/44)]、肝功能损伤[22.22％(10/45)vs.15.91％(7/44)]、消化道症状[60.00％ (27/45)vs.70.45％(31/44)]、肾功能损伤[8.89％ (4/45)vs.13.64％ (6/44)]发生率差异均无统计学意义(P＞0.05).对照组患者的总生存时间中位数为23.988个月,观察组患者的总生存时间中位数为27.976个月,两组差异有统计学意义(x2=4.177,P＜0.05).结论 使用腹腔镜微波消融术联合三维适形放疗治疗肝癌转移患者,可以提升治疗效果,改善生活质量,延长患者的生存时间,安全性良好,值得临床推荐.

Objective To compare the safety and short-term efficacy between uncut Roux-en-Y anastomosis and traditional Roux-en-Y anastomosis in digestive tract reconstruction after totally laparoscopic distal gastrectomy. Methods Retrospective analyzed the clinical data of 43 patients with total laparoscopic radical gastrectomy for gastric cancer between January 2015 and June 2016(23 patients with uncut Roux-en-Y anastomosis,20 patients underwent conventional Roux-en-Y anastomosis),and the perioperative related indicators and postoperative follow-up of the two groups were analyzed retrospectively. Results Compared with group B,group A had shorter time in digestive tract reconstruction and less blood loss(P<0.01). The operation time of group A was shorter than that of group B(P<0.05). After 6 months follow-up,none of the two groups died, and the incidence of postoperative complications was similar(P>0.05). Conclusion Uncut Roux-en-y digestive tract anastomosis after laparoscopic distal gastrectomy for gastric cancer has the advantages of short operation time and low bleeding volume,and does not increase the incidence of postoperative adverse reaction,and it is worth clinical application.

目的:尝试多种化疗药物联合使用来诱导人肝癌Bel-7402细胞株,得到具有多药耐药性的人肝癌细胞株,测定细胞的生物学特性,并与亲本细胞株和单药诱导得到多药耐药细胞株作比较. 方法:细胞分为A、B、C共3组,以人肝癌Bel-7402细胞为亲本细胞作为A组,B组利用大剂量顺铂(DDP)冲击法诱导建立多药耐药细胞株Bel-7402/DDP,C组利用大剂量奥沙利铂、5-氟尿嘧啶、 顺铂共同冲击诱导建立人肝癌多药耐药细胞株Bel-7402/X. 光镜下观察各组细胞形态学变化,CCK-8法测定3组细胞的生长曲线、 药物敏感性,细胞划痕实验比较各组细胞的迁徙能力. 结果:利用多种化疗药共同诱导成功得到多药耐药Bel-7402/X细胞株模型. 它和单药诱导得到的Bel-7402/DDP细胞株模型一样, 对多种化疗药物产生耐药性.Bel-7402/X细胞株的迁徙能力强于Bel-7402/DDP和Bel-7402细胞株,增殖活性小于Bel-7402/DDP和Bel-7402细胞株. 结论:Bel-7402/X细胞株对多种化疗药物有耐药性,可以用作逆转肿瘤多药耐药性体外实验的模型.

Objective: The aim of this study was to analyse the shunt placement in patients who had normal pressure hydrocephalus after poor-grade aneurysmal subarachnoid haemorrhage (aSAH).Methods: Patients diagnosed with NPH after poor-grade aSAH were divided into a treatment group and control group, based on whether they had received ventriculoperitoneal shunt placement. The treatment group was then divided into an improvement group and non-improvement group according to their recovery. The Glasgow Outcome Scale and Mini Mental Scale Examination were used for 3 month and 1 year follow-up rehabilitation measures.Results: Of the 46 total patients, significant improvement was observed at the 3 month and 1 year follow-ups (p<0.01) after shunt implantation in the treatment group compared to the control group. Furthermore, patients who were younger (p = 0.022), had better neurological function (higher Glasgow Coma Score, p<0.01) and less severe hydrocephalus (lower EI, p<0.01) appears to be more likely to benefit from the shunt.Conclusions: Patients who had NPH due to poor-grade aSAH would benefit from shunt placement when given the correct candidates and timely management of shunt malfunction. Additionally, the curative effect of the shunt should have been regarded as a long-term goal of rehabilitation in these patients.

目的:探讨胰十二指肠同源盒基因-1 (Pdx1)在诱导多潜能干细胞(iPSCs)分化为胰岛β细胞中的作用及其机制.方法:体外培养人皮肤成纤维细胞来源的iPSCs,将该iPSCs定向诱导分化20 d;RT-PCR检测胰岛素相关基因的表达情况;比较诱导前后几个重要转录因子的表达情况;ChIP检测胰岛β细胞发育过程中最重要的转录因子Pdx1结合启动子区域的具体位点.结果:体外成功培养人皮肤来源的iPSCs;胰岛素相关基因MafA、insulin、Glut2、Nkx6.1、Glucokinase和Tcf1均呈现不同程度的表达增强,并在第20 d时表达基本达到高峰;实验组转录因子Pdx1、Ngn3与Pax6表达较对照组明显增强;Pdx1通过结合Insulin-P、Ngn3-P、Pax6-P区域激活下游基因Ngn3和Pax6.结论:Pdx1激活下游基因Ngn3和Pax6可能是其促进iPSCs定向诱导分化为胰岛β细胞的机制之一.

Objective: To investigate the mechanism of adipose tissue wound healing(ATWH). Methods: 3T3-L1 preadipocytes were cultured and expression of adiponectin receptors(AdipoR1/2)in 3T3-L1 preadipocytes was detected by immunohistochemistry and reverse transcription-polymerase chain reaction(RT-PCR). The concentrations of adiponectin secreted by 3T3-L1 preadipocytes were measured by ELISA. Proliferation and migration of 3T3-L1 preadipocytes were performed in vitro by MTT and wound closure assay, respectively. Results: AdipoR1/2 were expressed in 3T3-L1 preadipocytes. There were no statistically significant differences of adiponectin secreted by 3T3-L1 preadipocytes of different densities(P0.05). Adiponectin can promote 3T3-L1 preadipocytes proliferation and migration. Conclusions: Adiponectin might regulate ATWH by promoting preadipocytes proliferation and migration, and application of adiponectin systemically and/or locally is proposed to be a hopeful therapeutic approach for coping with surgical incision.

Objective To investigate relationship between adiponectin and adipocyte apoptosis during adipose tissue wound healing (ATWH),and clarify the role and its mechanism of adiponectin in subcutaneous suture to promote ATWH.Methods The pig models of ATWH were established to detect adiponectin expression of local adipose tissue by reverse transcriptase-polymerase chain reaction (RT-PCR),and apoptosis was examined by fluorescent labeling technique.3T3-L1 preadipocytes of mice were cultured in vitro.In experimental group 3T3-L1 preadipocytes were cultured with different concentrations of adiponectin,and apoptosis rate was tested by Annexin-V-FITC method.Results The expression of adiponectin in subcutaneous suture was significantly higher than that in control group (P ＜ 0.05).After 3 days and 5 days,the number of apoptotic cells was 29 ± 13 and 15 ± 8,respectively,statistically significantly less than that in the control group (69 ±25 and 23 ± 16) (P ＜0.05).With adiponectin concentration rising,the number of apoptotic cells was (26.42 ± 1.58) ％,(15.14 ± 1.91) ％,(7.04 ± 1.16) ％ and (5.05 ± 0.83) ％,significantly less than that in control group (33.18 ± 1.86) ％ (P ＜ 0.05).Conclusion Adiponectin plays a catalytic role in ATWH in the subcutaneous suture,and inhibiting adipocytes apoptosis is one of its mechanisms.