基于深度学习的肝脏病理图像中肝脂肪变性分级研究

摘要 目的：设计基于深层神经网络模型用来分析肝脏全景病理切片图像（Whole slide images, WSI）的肝脂肪变性分级方法,以实现对非酒精性脂肪性肝病（Non-alcoholic fatty liver disease, NAFLD）病程的辅助诊断。方法：结合临床诊断,以非酒精性脂肪肝活动度积分（NAFLD activity score, NAS）为评价标准,将肝脂肪变性程度分为无、轻度、中度和重度等四级病程,本研究采用多示例学习的策略构建并训练深度神经网络模型,将训练获得的人工智能模型用来实现计算机自动化诊断肝脏病理切片中肝脂肪变性程度分级。结果：通过使用本研究中的人工智能方法可以在3分钟内对一张WSI进行完整的分析,得到该病患肝脏病理切片中肝脂肪变性分级,训练获得的人工智能模型的AUC为0.97,肝脂肪变性分级的平均准确率为78.18%,macro-F1 score、macro-Precision和macro-Recall分别为79.49、82.03和77.10,其结果展示获得的人工智能模型已满足可辅助临床诊断的水平。结论：本研究基于深度学习技术开发的人工智能方法初步实现快速自动化诊断肝脂肪变性分级,展现了其潜在的临床使用价值。     

超声E成像在非酒精性脂肪性肝病中的诊断价值研究

摘要 目的：研究超声E成像在非酒精性脂肪性肝病中的诊断价值。方法：选取2018年1月-2020年2月我院诊断的非酒精性脂肪性肝病患者120例作为观察组,另外,选取同期健康体检的志愿者101例作为对照组,分别对两组受试者进行超声E成像、MR mDixon序列检查,比较两组受试者以及不同病情严重程度患者的谷丙转氨酶（ALT）、谷草转氨酶（AST）、总胆红素（TBIL）、碱性磷酸酶（ALP）、杨氏模量值（ElastPQ）以及脂肪含量之间的差异,分析患者的病情严重程度与ElastPQ以及脂肪含量的相关性。结果：观察组的ALT、AST、TBIL、ALP、ElastPQ以及脂肪含量显著高于对照组（P＜0.05）;不同病情严重程度患者的ALT、AST、TBIL、ALP、ElastPQ以及脂肪含量之间的差异有统计学意义（P＜0.05）,ALT、AST、TBIL、ALP、ElastPQ以及脂肪含量从高到低依次为重度组、中度组以及轻度组,患者的病情严重程度与ElastPQ以及脂肪含量呈正相关（P＜0.05）,ElastPQ与脂肪含量呈正相关（P＜0.05）。结论：超声E成像对于非酒精性脂肪性肝病患者肝脏组织的受损程度具有较高的诊断价值,建议临床推广应用。     

恩格列净对2型糖尿病合并非酒精性脂肪性肝病的影响

摘要 目的：探讨恩格列净单独及联合胰岛素治疗对2型糖尿病合并非酒精性脂肪性肝病（NAFLD）患者的影响。方法：本研究选择2021.3-2022.8于石家庄市第二医院门诊就诊的2型糖尿病合并NAFLD的300例患者作为研究对象,分为胰岛素治疗组（100例）、恩格列净单药治疗组（100例）及恩格列净联合胰岛素治疗组（100例）,治疗48周,采集患者治疗前后体重、体质指数（BMI）、谷草转氨酶（AST）、谷丙转氨酶（ALT）、谷酰转肽酶（GGT）、血常规、HbA1c、肝脏脂肪含量等指标,计算患者NFS、FIB-4指标,比较不同治疗方案治疗前后患者临床特征的变化。观察三组临床治疗期间不良反应的发生情况。结果：应用胰岛素治疗的患者HbA1c及空腹血糖显著下降;BMI、ALT、AST、GGT、肝脏脂肪含量、NFS、FIB-4无变化。恩格列净单药治疗的患者,ALT、AST、GGT、NFS、FIB-4、肝脏脂肪含量均显著下降（P<0.05）;恩格列净联合胰岛素治疗的患者ALT、AST、GGT、NFS、FIB-4、肝脏脂肪含量显著下降（P<0.05）。多元线性回归提示应用恩格列净和肝脏脂肪含量的变化相关（P<0.05）,BMI、HbA1c的变化和肝脏脂肪含量的变化无关（P<0.05）。三组患者治疗期间总不良反应发生率差异无统计学意义（P＞0.05）。结论：恩格列净单药治疗或联合胰岛素治疗均可降低2型糖尿病合并NAFLD患者肝脏脂肪含量均、转氨酶水平,改善肝纤维化。     

基于肝病十二分级诊疗系统的酒精性肝硬化临床路径研究

目的 根据医院自行开发的肝病十二分级诊断系统研制酒精性肝硬化疾病的临床路径。方法 根据帕累托20/80准则，对306例酒精性肝硬化的47个医嘱项目进行频数统计，然后根据肝病十二分级诊断情况再进行帕累托判定。结果 共有35个医嘱项目进入酒精性肝硬化的临床路径。结论 依据肝病十二分级诊断系统对患者进行分类、分期、分队列之后与医嘱项目进行联动，能够制定较为合理的酒精性肝硬化临床路径。     

Gdf15对酒精性脂肪肝代谢异常的影响

摘要 目的：探讨生长分化因子15（Growth and Differentiation Factor 15, GDF15）对于酒精性脂肪肝(alcoholic fatty liver, AFL)代谢异常的影响。方法：用白酒构建酒精性脂肪肝小鼠模型然后分别给对照组和模型组尾静脉注射AAV8-GDF15过表达肝脏的GDF15分子,将小鼠共分为四组：正常+尾静脉注射对照AAV8-NC组(Con+AAV8-NC)、模型(酒精)+尾静脉注射AAV8对照病毒组(AFL+AAV8-NC)、正常+尾静脉注射过表达AAV8-Gdf15组(Con+AAV8-Gdf15)、酒精+尾静脉注射过表达AAV8-Gdf15组(AFL+AAV8-Gdf15)。对其体重、空腹血糖、葡萄糖耐量、胰岛素释放、血清脂、肝脏脂、谷丙转氨酶、谷草转氨酶含量的测定;其肝脏活组织切片使用苏木精-伊红染色法染色检测肝脏结构异常;q-PCR检测脂代谢相关分子RNA水平等观察过表达GDF15对酒精性脂肪肝的影响。结果：与Con+AAV8-N组相比,AFL+AAV8-NC组的体重下降,而过表达GDF15后AFL+AAV8-Gdf15组体重比AFL+AAV8-NC组的体重下降减少。与Con+AAV8-NC组相比,AFL+AAV8-NC组的空腹血糖升高、糖耐量及胰岛素耐量下降,过表达GDF15后AFL+AAV8-Gdf15组与AFL+AAV8-NC相比空腹血糖显著下降、糖耐量及胰岛素耐量显著升高。AFL+AAV8-NC组与Con+AAV8-NC组相比血脂TG明显升高,过表达GDF15后AFL+AAV8-Gdf15组血脂与AFL+AAV8-NC的血脂相比显著下降。与Con+AAV8-NC组相比,AFL+AAV8-NC组的肝脏重量增加,肝功能损伤程度更严重,肝脏脂肪含量增加,而过表达GDF15后AFL+AAV8-Gdf15与AFL+AAV8-NC组相比,肝脏重量、损伤程度及肝脏的脂肪含量均有显著性下降。结论：酒精性脂肪肝增加GDF15的表达,而GDF15的表达增加会改善酒精性脂肪肝的损伤及代谢异常。     

非酒精性脂肪肝与脂肪性肝炎动物模型研究进展

非酒精性脂肪肝是无酒精滥用的包括单纯性脂肪肝、脂肪性肝炎、脂肪性肝纤维化和肝硬化的肝病综合征,目前已成为广受关注的肝病医学难题。随着抗脂肪肝药物的深入研究,动物模型制作得到很好发展。近年来,在大鼠、沙鼠、小鼠、兔和小猪等动物种属成功地建立了食物、胃肠外营养与蛋氨酸胆碱缺乏等诱导的单纯性脂肪肝和脂肪性肝炎动物模型,这些模型为研究脂肪肝和脂肪性肝炎的发病机理与治疗提供了机会。每种动物模型各有优缺点,合理应用动物模型能更好地开展脂肪肝病的实验和临床研究。本文综述了非酒精性脂肪肝及脂肪性肝炎动物模型制作方法的若干研究进展。     

乙酰辅酶A羧化酶抑制剂联合非诺贝特对小鼠非酒精性脂肪肝的治疗效果

摘要 目的：探讨乙酰辅酶A羧化酶抑制剂（MK-4074）联合非诺贝特对小鼠非酒精性脂肪肝（NAFLD）的脂质含量以及肝功能的改善效果。方法：20只C57BL/6小鼠给予60%高脂饲料连续喂养8周构建NAFLD小鼠模型后，随机分为安慰剂组、MK-4074组、非诺贝特组以及MK-4074联合非诺贝特治疗组，每组各5只，继续高脂喂养并分别给予安慰剂（Placebo）、MK-4074（10 mg/kg/天）、非诺贝特（30 mg/kg/天）、以及MK-4074（10 mg/kg/天）+ 非诺贝特（30 mg/kg/天）治疗持续8周。治疗结束后对小鼠体重、肝指数、肝脏脂质含量、肝功能以及肝脏病理和肝脏中性粒细胞和巨噬细胞浸润情况进行分析。结果：与安慰剂组相比，单用MK-4074治疗可显著降低肝指数、肝脏甘油三酯（TG）、胆固醇（TC）、非酯化脂肪酸（NEFA）的含量以及血清ALT和AST水平，而对小鼠体重和血清TC没有显著影响；单用非诺贝特可显著降低小鼠体重，肝脏TG、TC、NEFA以及血清TG、 ALT和AST水平，对小鼠的肝指数、血清TC没有显著影响；而MK-4074与非诺贝特联合治疗可显著降低小鼠体重、肝脏TG、TC、NEFA，以及血清TG、ALT和AST水平，降低肝脏脂质积累以及中性粒细胞与巨噬细胞浸润，效果优于MK-4074或非诺贝特单药治疗。结论：MK-4074联合非诺贝特可显著减少NAFLD小鼠肝脏的脂质含量，改善肝功能。     

基于X线图像的膝关节周围原发性骨肿瘤辅助诊断的机器学习模型研究

摘要 目的：开发机器学习模型,并评估其在膝关节周围原发性骨肿瘤诊断方面的准确性。方法：本文将深度卷积神经网络（DCNN）这一深度学习方法应用于膝关节X线图像的影像组学分析,探讨其辅助诊断膝关节周围原发性骨肿瘤的临床价值。结果：该深度学习模型在区分正常与肿瘤影像方面展现出优异的诊断准确性,使用DCNN模型进行5轮测试的总体准确性为（99.8±0.4）%,而阳性预测值和阴性预测值分别为（100.0±0.0）%和（99.6±0.8）%,各个数据集的曲线下面积（AUC）分别为0.99、1.00、1.00、1.0和1.0,平均AUC为（0.998±0.004）;进一步使用DCNN模型进行了10轮测试显示其在区分良性与恶性骨肿瘤方面的总体准确性为（71.2±1.6）%,且达到了强阳性预测值（91.9±8.5）%,各个数据集的AUC分别为0.63、0.63、0.58、0.69、0.55、0.63、0.54、0.57、0.73、0.63,平均AUC为（0.62±0.06）。结论：本文是首个将人工智能技术应用于骨肿瘤诊断的X线图像影像组学分析方面的研究,人工智能影像组学模型能够帮助医生自动地快速筛查骨肿瘤,确定良性或恶性肿瘤时,阳性预测值较高。     

肠-肝轴与非酒精性脂肪性肝病

肠道菌群组成和数量的改变影响宿主的能量代谢、免疫应答和炎症反应状态。非酒精性脂肪性肝病患者常伴有小肠细菌过度生长或某些菌群种类和数量的改变,以及肠道黏膜通透性增加。肠道细菌通过增强肝脏脂肪合成、诱导机体胰岛素抵抗、激活天然免疫系统相关分子模式等机制,诱发肝脏炎症反应,启动纤维化进程,促进单纯性脂肪变向脂肪性肝炎发展。鉴定影响机体能量代谢和炎症反应的肠道菌群及其产物将为阐明肠-肝轴对肝脏炎症发生、发展所起的作用奠定基础,为揭示非酒精性脂肪性肝病发生、发展的机制开辟新思路,为该病的防治探索新策略。     

肝血窦内皮细胞介导的他莫昔芬对非酒精性脂肪性肝炎治疗作用的转录组学分析

摘要 目的：探究他莫昔芬对饮食诱导的非酒精性脂肪性肝炎小鼠肝血窦内皮细胞的代谢、炎症及纤维化等通路基因表达的影响。方法：采用8周龄的雄性C57BL/6小鼠,给予MCD饲料喂养6周后,每天腹腔注射一次他莫昔芬（100 mg/kg）,持续5天。分离并收集肝血窦内皮细胞,加入1 mL TRIzol试剂吹打至沉淀消失,放入-80℃冰箱保存。样本后续送至基迪奥生物公司进行转录组测序并在Omicsmart平台进行生物信息学分析。部分生物信息学分析数据来自已经发表的文献并通过Omicshare分析平台分析。结果：转录组测序发现,差异基因KEGG和GO分析发现差异基因在免疫和炎症通路富集。通过分析肝脏内皮特异性代谢基因表达,我们发现他莫昔芬治疗逆转了NASH过程中部分代谢基因的下调,以及NASH过程中CCL2、CXCL2、CXCL5和VCAM-1等促炎基因和Col1a1、Col1a2、Col3a1、Tgfb2、和Timp1等促纤维化基因的表达上调。同时,GSEA分析也显示他莫昔芬抑制了炎症和纤维化通路的表达。结论：他莫昔芬可能通过逆转非酒精性脂肪性肝炎对小鼠肝血窦内皮细胞代谢基因的改变以及炎症及纤维化相关基因的上调来治疗非酒精性脂肪性肝炎。     

New scoring system combining the FIB-4 index and cytokeratin-18 fragments for predicting steatohepatitis and liver fibrosis in patients with nonalcoholic fatty liver disease

Purpose: To establish a new scoring system as a noninvasive tool for predicting steatohepatitis and liver fibrosis in patients with nonalcoholic fatty liver disease (NAFLD).

Methods: A total of 170 patients histologically diagnosed with nonalcoholic steatohepatitis (NASH) (n?=?130) or nonalcoholic fatty liver (NAFL) (n?=?40) were enrolled. We analyzed receiver operating characteristic (ROC) curves and performed multivariate analysis to predict steatohepatitis and liver fibrosis.

Results: Multivariate analysis showed that cytokeratin-18 fragment (CK18-F) levels (≥278?U/L) (odds ratio [OR], 4.46; 95% confidence interval [CI], 1.42–14.00; p?=?0.010) and the FIB-4 index (≥1.46) (OR, 4.54; 95% CI, 1.93–29.50; p?=?0.004) were independently associated with prediction of NASH. We then established a new scoring system (named the FIC-22 score) for predicting NASH using CK18-F levels and FIB-4 index. The areas under the ROC curve (AUROCs) of the FIC-22 score and NAFIC score were 0.82 (95% CI, 0.75–0.89) and 0.71 (95% CI, 0.62–0.78) (p?=?0.044). Additionally, the AUROC of the FIC-22 score for predicting the presence of fibrosis (F?≥?1) was 0.78 (95% CI, 0.70–0.85).

Conclusions: In patients with NAFLD, the FIC-22 score had high predictive accuracy not only for steatohepatitis but also for the presence of liver fibrosis.     

瞬时弹性成像技术对非酒精性单纯性脂肪肝的诊断价值及受控衰减系数与糖脂代谢的相关性分析

目的:探讨瞬时弹性成像技术(FibroTouch)对非酒精性单纯性脂肪肝(NAFLD)的诊断价值及受控衰减系数(CAPTM)与糖脂代谢的相关性分析。方法:选取2017年8月~2019年8月期间本院收治的NAFLD患者117例为研究对象,纳入实验组,另选同期在我院进行健康体检的健康受试者80例为对照组。对实验组患者和对照组受试者进行FibroTouch检查、腹部彩色多普勒超声检查、糖脂代谢指标生化检查。对比两组受试者的糖脂代谢指标水平、CAPTM水平、FibroTouch检查与彩色多普勒超声检查诊断NAFLD的准确率,分析CAPTM水平与糖脂代谢指标水平的相关性。结果:实验组空腹血糖(FPG)、糖化血红蛋白(HbA1C)、空腹胰岛素(FINS)水平均高于对照组(P<0.05)。实验组甘油三酯(TG)、总胆固醇(TC)、低密度脂蛋白(LDL-C)、载脂蛋白A(apoA)、CAPTM水平均高于对照组(P<0.05)。FibroTouch检查NAFLD的准确率高于彩色多普勒超声检查(P<0.05)。经Pearson相关性分析显示,CAPTM与各项糖脂代谢指标均呈正相关性(P<0.05)。结论:相比于彩色多普勒超声检查,FibroTouch用于NAFLD的诊断准确率更高,FibroTouch检查参数CAPTM与糖脂代谢指标存在明显的相关性,可辅助用于NAFLD的诊断和病情评估。     

Capsaicin suppresses liver fat accumulation in high-fat diet-induced NAFLD mice

ABSTRACT

Dietary capsaicin exhibits anti-steatosis activity in obese mice. High-fat diet (HFD)-induced mice is a highly studied approach to develop non-alcoholic fatty liver disease (NAFLD). In this study, we determined whether the topical application of capsaicin can improve lesions of NAFLD. The HFD-induced mice were treated with daily topical application of capsaicin for 8 weeks. Topical application of capsaicin reduced liver fat in HFD-fed mice. Capsaicin stimulated carnitine palmitoyl transferase (CPT)-1 and CD36 expression, which are associated with β-oxidation and fatty acids influx of liver while it decreased the expression of key enzymes involved in the synthesis of fatty acids, such as acetyl Co-A carboxylase (ACC) and fatty acid synthase (FAS). Immunohistochemical analysis revealed the elevated level of adiponectin in liver tissue of the capsaicin-treated mice. These results suggest that the topical application of capsaicin suppresses liver fat accumulation through the upregulation of β-oxidation and de novo lipogenesis in HFD-induced NAFLD mice.     

A high-fat and cholesterol diet causes fatty liver in guinea pigs. The role of iron and oxidative damage

Abstract

Non-alcoholic fatty liver disease (NAFLD) is a common chronic liver disease. Iron, cholesterol, and oxidative damage are frequently suggested to be related to the progression of NAFLD, but the precise relationship between them remains unclear. Guinea pigs fed on a high cholesterol and fat diet (without oxidized lipids) generated a disease model of NAFLD with hallmark observations in liver histology and increased liver damage markers. Hepatic cholesterol and iron levels were found to be significantly elevated and directly correlated. Plasma hepcidin and transferrin levels were decreased. Plasma iron concentrations were found to be elevated, likely due to an increased intestinal iron absorption caused by the decrease in plasma hepcidin. However, hepatic transferrin receptor-2 levels were unchanged. No significant increase in hepatic lipid peroxidation was detected using F₂-isoprostanes as a reliable biomarker, nor was there a rise in protein carbonyls, a general index of oxidative protein damage. Some increases in cholesterol oxidation products were observed, but largely negated after normalizing for the elevated hepatic cholesterol content. Indeed, increased hemosiderin deposition and unchanged ferritin levels in liver suggested that the excess iron mainly existed as hemosiderin, which is redox-inactive.     

Berberine reduces methylation of the MTTP promoter and alleviates fatty liver induced by a high-fat diet in rats

High-calorie food leads to nonalcoholic fatty liver disease (NAFLD) through dysregulation of genes involved in lipid metabolism, but the precise mechanism remains unclear. DNA methylation represents one of the mechanisms that contributes to dysregulation of gene expression via interaction with environmental factors. Berberine can alleviate fatty liver in db/db and ob/ob mice. Here, we investigated whether DNA methylation is involved in the pathogenesis of NAFLD induced by a high-fat diet (HFD) and whether berberine improves NAFLD through influencing the methylation status of promoters of key genes. HFD markedly decreased the mRNA levels encoding CPT-1α, MTTP, and LDLR in the liver. In parallel, DNA methylation levels in the MTTP promoter of rats with NAFLD were elevated in the liver. Interestingly, berberine reversed the downregulated expression of these genes and selectively inhibited HFD-induced increase in the methylation of MTTP. Consistently, berberine increased hepatic triglyceride (TG) export and ameliorated HFD-induced fatty liver. Furthermore, a close negative correlation was observed between the MTTP expression and its DNA methylation (at sites −113 and −20). These data indicate that DNA methylation of the MTTP promoter likely contributes to its downregulation during HFD-induced NAFLD and, further, that berberine can partially counteract the HFD-elicited dysregulation of MTTP by reversing the methylation state of its promoter, leading to reduced hepatic fat content.     

Fatty liver induced by free radicals and lipid peroxidation

Abstract

An excessive accumulation of fat in the liver leads to chronic liver injury such as non-alcoholic fatty liver disease (NAFLD), which is an important medical problem affecting many populations worldwide. Oxidative stress has been implicated in the pathogenesis of NAFLD, but the exact nature of active species and the underlying mechanisms have not been elucidated. It was previously found that the administration of free radical-generating azo compound to mice induced accumulation of fat droplet in the liver. The present study was performed aiming at elucidating the changes of lipid classes and fatty acid composition and also measuring the levels of lipid peroxidation products in the liver induced by azo compound administration to mouse. The effects of azo compound on the liver were compared with those induced by high fat diet, a well-established cause of NAFLD. Azo compounds given to mice either by intraperitoneal administration or by dissolving to drinking water induced triacylglycerol (TG) increase and concomitant phospholipid decrease in the liver, whose pattern was quite similar to that induced by high fat diet. Lipid peroxidation products such as hydroxyoctadecadienoic acid and hydroxyeicosatetraenoic acid were increased in the liver in association with the increase in TG. These results show that free radicals as well as high fat diet induce fatty liver by similar mechanisms, in which lipid peroxidation may be involved.     

泽泻汤提取物三萜类化学成分分析及其抗非酒精性脂肪肝作用研究

目的:对泽泻汤不同极性溶剂提取物三萜类化学成分进行定性分析并探究其对非酒精性脂肪肝的干预作用。方法:制备泽泻汤水及95%乙醇提取物,利用超高效液相色谱串联高分辨质谱技术对三萜类成分进行定性鉴别,并比较两种提取物中三萜类化合物数量及含量差异。采用油酸诱导的HepG2细胞脂肪累积模型评价泽泻汤不同极性提取物干预非酒精性脂肪肝的能力。结果:在泽泻汤中共鉴定20个三萜类色谱峰,水提取物中鉴定出14个三萜类成分,其中16-oxo-alisol A相对含量最高,95%乙醇提取物中鉴定出16个三萜类成分,alisol B 23-acetate相对含量最高。两提取物均能显著降低细胞中的脂肪累积量,且乙醇提取物表现出更强的抑制细胞内脂肪累积的作用。结论:泽泻汤水及95%乙醇提取物中三萜类化合物数量和含量存在差异,两种提取物均具有潜在抗非酒精性脂肪肝作用,且乙醇提取物活性更强,研究结果为泽泻汤进一步的药效物质基础及药理活性研究提供了基础。     

NAFLD患者肠粘膜机械屏障损伤的研究

非酒精性脂肪性肝病(nonalcoholic fatty liver disease,NAFLD)是慢性肝损伤的主要病因之一。据估计,大约有20%的成人有非酒精性脂肪性肝病,2%-3%发展成非酒精性脂肪性肝炎(nonalcoholic steatohepatitis,NASH)。NASH是NAFLD的渐进形式,并可能导致肝硬化和肝细胞癌。NAFLD不仅增加了肝病患者死亡率,作为代谢综合征,还增加了肥胖、2型糖尿病及高脂血症的发病率。肌球蛋白轻链激酶(MLCK)是细胞收缩的关键酶,可使肌球蛋白轻链磷酸化(MLC),促使肌动蛋白收缩,破坏细胞间的紧密连接蛋白,使细胞骨架收缩,进而使肠上皮通透性增加,肠粘膜机械屏障遭到破坏,致使NAFLD的病情进一步发展。MLCK在NAFLD的发生及发展中起着重要作用。NAFLD严重威胁人类健康,影响人们的生活质量及生存质量。为NAFLD患者寻找崭新的治疗方法是极其必要的。