Celastrus orbiculatus Extracts Inhibit Human Hepatocellular Carcinoma Growth by Targeting mTOR Signaling Pathways

Objective: To characterize the molecular mechanism underlying the antineoplastic activity of Celastrus orbiculatus Thunb. extracts(COE). Methods: The human hepatocellular carcinoma HepG2 cells with mammalian target of rapamycin(mT OR) knockdown expressed(HepG 2/mT OR-) were constructed using molecular biological technology. In vitro, the HepG 2/mT OR-cells were treated with COE at various concentrations(10, 20, 40, 80, 160 and 320 μg/mL). Cell viability was determined using 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide(MTT) assays. According to the half-maximal inhibitory concentration(IC50) value(140 mg/L), the concentrations of COE in the subsequent experiment was set to alleviate cytotoxicity. The HepG 2/mT OR-cells were divided into 5 groups: negative control(untreated), COE treatment groups(40, 80, 120 mg/L COE) and positive control group(cisplatin, DDP, 2 mg/L), respectively. Wild-type HepG 2 cells were used as a blank control. The effects of COE on the cell apoptosis were analyzed by flow cytometry and transmission electronic microscopy(TEM), respectively. The protein expression levels of mT OR signal pathways were determined by Western blotting. In vivo, HepG2/mTOR-cells(2×10~6 cell/mice) were subcutaneously injected into the right flank of nude mice. Thirty-six female nude mice were randomly assigned to 6 groups according to body weight(6 mice per group) as follows: solvent vehicle control, Banmao Capsule treated group(BM, 195 mg/kg), Tegafur, Gimeracil and Oteracil Potassium Capsules(10 mg/kg) treated group, and different dosages of COE(10, 20, 40 mg/kg) groups. Tumor growth was monitored and immunohistochemical staining was used to examine the expression of apoptosis-related proteins in tumor tissues. Results: COE inhibited the proliferation significantly in a concentration-dependent manner in HepG 2/mT OR-cells(P0.01). COE significantly induced the apoptosis of HepG 2/mT OR-cells(P0.01), and the apoptotic bodies can be observed under TEM. COE significantly inhibits the proteins expression of mT ORrelated signal pathways. In vivo, COE significantly inhibited tumor growth in nude mice(P0.01). Moreover, the results showed that COE down-regulated the expression of Bcl-2 and Bcl-xL, and up-regulated the levels of Bax and caspase-3 protein(P0.01). Conclusion: COE was a potential chemotherapeutic drug in HCC treatments via targeting mT OR signal pathway.     

The role of lncRNA MSC-AS1/miR-29b-3p axis-mediated CDK14 modulation in pancreatic cancer proliferation and Gemcitabine-induced apoptosis

Pancreatic ductal adenocarcinoma (PDAC) remains a leading cause of cancer-related death due to the failure of traditional therapies. In the present study, we attempted to construct a lncRNA-miRNA-mRNA network which may modulate PDAC cell proliferation and Gemcitabine-induced cell apoptosis starting from CDK14, a new member of the CDK family and an oncogene in many cancers. Based on TCGA data, a significant positive correlation was observed between lncRNA MSC-AS1 and CDK14. Moreover, MSC-AS1 expression was upregulated in PDAC tissues. Higher MSC-AS1 expression was correlated with poorer prognosis in patients with PDAC. MSC-AS1 knockdown in Panc-1 and BxPC-3 cells significantly inhibited the cell proliferation. Moreover, miR-29b-3p, which has been reported to act as a tumor suppressor, was predicted to bind to both MSC-AS1 and CDK14. Contrary to MSC-AS1, higher miR-29b-3p expression was correlated to better prognosis in patients with PDAC. In both PDAC cell lines, miR-29b-3p negatively regulated MSC-AS1 and CDK14. As confirmed using luciferase reporter gene and RIP assays, MSC-AS1 served as a ceRNA for miR-29b-3p to counteract miR-29b-mediated CDK14 repression. MSC-AS1 knockdown inhibited CDK14 protein levels and PDAC proliferation and enhanced gemcitabine-induced cell death and apoptosis while miR-29b-3p inhibition exerted an opposing effect; the effect of MSC-AS1 knockdown was partially attenuated by miR-29b-3p inhibition. Taken together, we demonstrated that MSC-AS1/miR-29b-3p axis modulates the cell proliferation and GEM-induced cell apoptosis in PDAC cell lines through CDK14. We provided a novel experimental basis for PDAC treatment from the perspective of lncRNA-miRNA-mRNA network.     

Mitochondrial dysfunction in affected skin and increased mitochondrial DNA in serum from patients with psoriasis

Psoriasis is characterized by keratinocyte proliferation and chronic inflammation, but the pathogenesis is still unclear. Dysregulated mitochondria (mt) could lead to reduced apoptosis and extracellular secretion of mtDNA, acting as “innate pathogen” triggering inflammation. Serum was obtained from healthy volunteers and psoriatic patients. Mitochondrial DNA was extracted from the serum and amplified with quantitative PCR (qPCR). Punch biopsies were obtained from lesional and non‐lesional psoriatic skin (10 cm apart) and from healthy volunteers, were placed in RNA later and were stored at ?80°C until RNA was extracted and cDNA was synthesized; gene expression of uncoupling protein 2 (UCP2), Dynamin‐related protein 1 (Drp1) and calcineurin, involved in the regulation of mitochondria function, was detected with qPCR. Mitochondrial DNA was significantly increased (7s, P = 0.0496 and Cytochrome B, CytB, P = 0.0403) in the serum of psoriatic patients (n = 63) as compared to controls (n = 27). Gene expression was significantly reduced for UCP2 (P = 0.0218), Drp1 (P = 0.0001) and calcineurin (P = 0.0001) in lesional psoriatic skin, as compared to non‐lesional or control skin. Increased serum extracellular mtDNA in psoriatic patients and decreased expression of mitochondrial regulatory proteins in psoriatic skin suggest increased inflammation and reduced keratinocyte apoptosis, respectively. Inhibitors of mtDNA secretion and/or UCP2 stimulants may be potential treatment options.     

直肠癌淋巴结转移的深度神经网络评判研究进展

正目前,临床上对于直肠癌常用的影像评估方法有MRI、螺旋CT、PET-CT、直肠腔内超声(ERUS)等。而MRI作为首选检查方式,对肿瘤位置、浸润深度、淋巴结转移、血管侵犯、环周切缘及周围器官侵犯等方面的评估均具有明显优势~([1-2])。通过MRI诊断淋巴结的方法通常是影像科医师逐层浏览每一幅图像,从中识别淋巴结的形状、界限及密度来判断,这种传统方式耗时较长且存在主观偏倚,导致     

靶向抑制ZNF281基因表达对直肠癌细胞生长抑制及化疗增敏作用的机 制探讨

目的:探讨抑制ZNF281基因表达对直肠癌细胞活力、凋亡率及5-氟尿嘧啶(5-FU)化疗敏感性的影响。方法:以LipofectamineTM 2000（Lipo2000）为载体，将siRNA转染人直肠癌Colo320细胞，Colo320细胞分为NC组（阴性对照siRNA）、si-ZNF281组（靶向抑制ZNF281表达的小干扰RNA）和si-ZNF281+5-FU组，MTT检测细胞活力，流式细胞仪检测细胞凋亡率，Western blotting检测β-catenin、PCNA和Survivin的蛋白表达。结果:ZNF281 siRNA转染Colo320细胞后，ZNF281表达显著低于空白组（P<0.05）。与NC组比较，si-ZNF281组细胞活力降低，凋亡率升高，β-catenin、PCNA和Survivin的蛋白表达降低，差异均具有统计学意义（P<0.05）。与si-ZNF281组比较，si-ZNF281+5-FU组细胞活力降低，凋亡率升高，β-catenin、PCNA和Survivin的蛋白表达降低，差异均具有统计学意义（P<0.05）。结论:抑制ZNF281基因表达可降低直肠癌细胞活力及诱导细胞凋亡，且可增加5-FU化疗敏感性，机制与抑制Wnt/β-catenin信号通路有关。     

Computational pathology definitions,best practices,and recommendations for regulatory guidance: a white paper from the Digital Pathology Association

In this white paper, experts from the Digital Pathology Association (DPA) define terminology and concepts in the emerging field of computational pathology, with a focus on its application to histology images analyzed together with their associated patient data to extract information. This review offers a historical perspective and describes the potential clinical benefits from research and applications in this field, as well as significant obstacles to adoption. Best practices for implementing computational pathology workflows are presented. These include infrastructure considerations, acquisition of training data, quality assessments, as well as regulatory, ethical, and cyber-security concerns. Recommendations are provided for regulators, vendors, and computational pathology practitioners in order to facilitate progress in the field. © 2019 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of Pathological Society of Great Britain and Ireland.     

Ginsenoside Rg1 prevents acetaminophen-induced oxidative stress and apoptosis via Nrf2/ARE signaling pathway

Inappropriate use of acetaminophen (APAP) can lead to morbidity and mortality secondary to hepatic necrosis. Ginsenoside Rg1 is a major active ingredient in processed Panax ginseng, which is proved to elicit biological effects. We hypothesized the beneficial effect of Rg1 on APAP-mediated hepatotoxicity was through Nrf2/ARE pathway. The study was conducted in cells and mice, comparing the actions of Rg1. Rg1 significantly improved cell survival rates and promoted the expression of antioxidant proteins. Meanwhile, Rg1 reduced the excessive ROS and the occurrence of cell apoptosis, which were related to Nrf2/ARE pathway. Expression of Nrf2 has a certain cell specificity.