液相色谱串联质谱法检测肝癌细胞N6- 甲基腺嘌呤核苷甲基化水平

目的 建立同时测定N⁶-甲基腺嘌呤核苷(m⁶A)和腺嘌呤核苷(A)的液相色谱-串联质谱法(LC-MS/MS),检测肝癌细胞m⁶A甲基化水平。方法 HepG₂和L₀₂细胞mRNA经分离、消化为核苷,再经含对乙酰氨基酚为内标的甲醇沉淀处理。色谱柱为Agilent Proshell 120 EC-C₁₈柱,流动相为0.1 g/dl甲酸水-甲醇(82:18),流速0.4 ml/min,柱温为30 ℃。质谱检测模式为多反应监测(DMRM)模式,测定m⁶A和A的浓度,计算细胞m⁶A甲基化水平。 结果 建立的LC-MS/MS法检测m⁶A和A的浓度分别在0.15～50.00 ng/ml和1.50～500.00 ng/ml浓度范围内线性关系良好(r > 0.999),日内、日间精密度均小于15.00 %,准确度为93.67 %～101.10 %,回收率为91.46 %～97.60 %,基质效应为90.26 %～99.27 %,样品稳定性良好。HepG₂和L₀₂细胞m⁶A甲基化水平分别为(0.73 ± 0.11)%和(1.26 ± 0.22)%。结论 该方法准确、快速、稳定和灵敏,可用于检测肝癌细胞m⁶A甲基化水平。     

Liver transplantation for intrahepatic and perihilar cholangiocarcinoma: Current and future

Cholangiocarcinomas are categorized as intrahepatic cholangio- carcinoma (iCCA), perihilar cholangiocarcinoma (pCCA), and dis- tal cholangiocarcinoma (dCCA) [1] . iCCA and pCCA are the second most common primary hepatobiliary malignant neoplasms [ 1 , 2 ]. Most patients with iCCA and pCCA have asymptomatic clinical course, highly aggressive nature and dismal prognosis [3] . Com- plete surgical resection offers the best possibility of long-term sur- vival but only a minority of patients are amenable to R0 resec- tion [4] . Tumor involvement of segmental bilateral intrahepatic bile duct, inadequate remnant liver volume or hepatic functional reserve due to underlying chronic liver disease, are key factors limiting the resectability of iCCA and pCCA. Liver transplantation (LT) provides an option for patients with unresectable iCCA and pCCA [4] .     

Damage Control Surgery for Severe Abdominal Sepsis With Free Skin Graft Transplantation: A Case Report

BackgroundAcute abdomen represents a great surgical challenge. Damage control surgery has been a better approach in acute abdominal emergencies to reduce the risk of mortality compared with primary definitive surgery.Case PresentationA 52-year-old man presented with nonspecific abdominal pain for 2 days. Physical examination and computed tomography scan suggested abdominal viscera perforation and acute diffuse peritonitis. Segmental resections and sigmoidostomy were performed, the abdomen was temporarily closed with a 3-liter infusion bag and maintained for 72 hours. Small intestinal ischemic necrosis was found, partial resection of small intestine was performed later, and abdomen was still closed with the 3-liter infusion bag and maintained for 10 days. One month later, intermediate split thickness self-free skin graft transplantation was performed. Abdomen was closed 6 months later.ConclusionsDamage control surgery with free skin graft transplantation can be successfully applied in severe abdominal infection in patients with an open abdomen. The study was in compliance with the Helsinki Congress and the Declaration of Istanbul.Core TipDamage control surgery has been a better approach in acute abdominal emergencies to reduce the risk of mortality compared with definitive surgery. Although laparostomy is a useful treatment of abdominal sepsis, successful management with free skin graft transplantation after laparostomy has rarely been reported in stercoral perforation. We report a patient with severe abdominal sepsis treated with damage control surgery with free skin graft transplantation. The patient was discharged with good recovery.     

如何从传统手术向智能化导航手术转变

经过20多年医理工多领域学科交叉融合、创新诊疗研发与临床实践, 数字医学4.0对传统手术的发展产生了深远而重要的影响。如何将传统外科手术与数字医学4.0技术相结合是未来外科学发展的方向。以数字智能化导航手术为代表的新技术在外科疾病的诊疗工作中得到了深入的探索和广泛的应用。随着人工智能、大数据、混合现实技术的创新发展及应用, 外科手术将会类似于航空航天自动化、智能化导航, 迎接数字医学5.0的到来。     

Decreased miR-124 contributes to the epithelial-mesenchymal transition phenotype formation of lung adenocarcinoma cells via targeting enhancer of zeste homolog 2

IntroductionMiR-124, a tumor suppressor, is involved in regulating various cellular processes. The purpose of this study was to investigate the possible function of miR-124 in LA (lung adenocarcinoma) cells.AimsMiR-124 expression levels in the 54 pairs of LA tissues (and corresponding non-tumor tissues) obtained at the Sixth People’s Hospital of Yancheng City and in LA cells were assessed by qRT-PCR. Colony formation assay, wound healing assay, transwell assays, attachment/detachment, western blotting and immunofluorescence assays were performed to assess the function of miR-124 on proliferation, migration and epithelial-to-mesenchymal (EMT) phenotypes in LA cells in vitro. Enhancer of zeste homolog 2 (EZH2) is identified as a target of miR-124 by bioinformatics analysis and luciferase reporter assays. Rescue assays were applied to verify the relationship between miR-124 and EZH2.ResultsMiR-124 was down-regulated in LA tissues (compared to adjacent non-tumor tissues), and was down-regulated in 3 out of 4 lung cancer cell lines compared to immortalized, non-tumorigenic bronchial epithelial cells. Forced expression of miR-124 significantly suppressed tumor cell proliferation, migration and inhibited the EMT process. On the contrary, deletion of miR-124 could obviously promote cell proliferation, migration and facilitate the formation of EMT phenotype. Bioinformatics analysis and luciferase reporter assays confirmed that EZH2 was a target gene of miR-124 and was negatively correlated with the level of miR-124 in cancer tissues.ConclusionOur current study suggested that miR-124 was a tumor suppressor in LA, and miR-124 was associated with LA cell EMT phenotype formation via targeting EZH2.