Geographic Variation of Intrahepatic Cholangiocarcinoma,Extrahepatic Cholangiocarcinoma,and Hepatocellular Carcinoma in the United States

Background

Intrahepatic (ICC) and extrahepatic cholangiocarcinomas (ECC) are tumors that arise from cholangiocytes in the bile duct, but ICCs are coded as primary liver cancers while ECCs are coded as biliary tract cancers. The etiology of these tumors is not well understood. It has been suggested that the etiology of ICC is more similar to that of another type of liver cancer, hepatocellular carcinoma (HCC), than to the etiology of ECC. If this is true, geographic incidence patterns and trends in ICC incidence should be more similar to that of HCC than ECC.

Methods

To examine this hypothesis, data from the North American Association of Central Cancer Registries Cancer in North America data file were analyzed. Incidence rates and joinpoint trends were calculated by demographic subgroup. County-level incidence rates were mapped.

Results

Overall incidence rates, racial distribution, male:female ratio, and peak ages were more similar between ICC and ECC than with HCC. During 2000–2009, average annual incidence rates of ECC increased. During 2005–2009, average annual ICC incidence rates also increased. High rates for all three cancer sites were found in the Pacific region, particularly Hawaii and Alaska. Rates of ICC and ECC were also high in the Northeast and the upper Midwest, while rates of HCC were high in the South.

Conclusions

Demographic patterns and geographical variation were more closely related between ICC and ECC than HCC, suggesting that the etiology of ICC and ECC may be similar. Increasing rates of both tumors suggest that further etiology studies are warranted.     

常规超声及超声造影在肝癌中的诊断及鉴别诊断价值

目的:探讨常规超声及超声造影在肝癌中的诊断及鉴别诊断价值。方法:收集2009年5月至2013年10月在我院进行住院诊治的肝癌患者120例,选择飞利浦IU22进行常规超声检查,选择飞利浦的IU22进行超声造影检查。结果:常规超声检查68例为低回声,20例为强回声,12例为等回声,17例为混合性回声,3例误诊为肝外病变,检出率为97.5%。肝癌组织的AT、TTP、ACT值明显晚于正常肝组织(P0.05),而BI、PI、MTT与BF值对比差异无明显差异(P0.05)。Spearman相关分析显示超声造影灌注参数AT、TTP、ACT与肿瘤分化程度呈正向相关性(P0.05)。结论:常规超声及超声造影在肝癌中的诊断及鉴别都有很好的应用价值,超声造影的应用可反映正常和病变组织的血流灌注情况,从而提高肝癌诊断的准确性。     

超声造影在肝硬化合并肝内局灶性小病灶中的诊断价值

目的:探讨超声造影(CEUS)在肝硬化合并肝内局灶性小病灶(≤3.0 cm)中的诊断价值。方法:选择我院自2010年4月至2014年8月患有肝硬化合并肝内局灶性小病灶的患者100例,采用MRI、CT及病理确定病灶良恶性,同时对比影像结果,计算常规超声和CEUS的准确性。结果:MRI、CT及病理确定病灶105个,其中恶性组45例,良性组60例,超声造影技术能清晰的显示出血流灌注的情况以及动脉相、门脉相及延迟相的特点。数据统计结果显示常规超声和CEUS的准确性、特异性和敏感性分别为59.05%、77.78%、45.00%和95.24%、95.56%、95.00%,两者之间的差异具有统计学意义(P0.05)。结论:相对于常规超声,CEUS对患有肝硬化合并肝内局灶性小病灶的患者具有更高的诊断价值。     

Mutation Inactivation of Nijmegen Breakage Syndrome Gene (NBS1) in Hepatocellular Carcinoma and Intrahepatic Cholangiocarcinoma

Nijmegen breakage syndrome (NBS) with NBS1 germ-line mutation is a human autosomal recessive disease characterized by genomic instability and enhanced cancer predisposition. The NBS1 gene codes for a protein, Nbs1(p95/Nibrin), involved in the processing/repair of DNA double-strand breaks. Hepatocellular carcinoma (HCC) is a complex and heterogeneous tumor with several genomic alterations. Recent studies have shown that heterozygous NBS1 mice exhibited a higher incidence of HCC than did wild-type mice. The objective of the present study is to assess whether NBS1 mutations play a role in the pathogenesis of human primary liver cancer, including HBV-associated HCC and intrahepatic cholangiocarcinoma (ICC). Eight missense NBS1 mutations were identified in six of 64 (9.4%) HCCs and two of 18 (11.1%) ICCs, whereas only one synonymous mutation was found in 89 control cases of cirrhosis and chronic hepatitis B. Analysis of the functional consequences of the identified NBS1 mutations in Mre11-binding domain showed loss of nuclear localization of Nbs1 partner Mre11, one of the hallmarks for Nbs1 deficiency, in one HCC and two ICCs with NBS1 mutations. Moreover, seven of the eight tumors with NBS1 mutations had at least one genetic alteration in the TP53 pathway, including TP53 mutation, MDM2 amplification, p14ARF homozygous deletion and promoter methylation, implying a synergistic effect of Nbs1 disruption and p53 inactivation. Our findings provide novel insight on the molecular pathogenesis of primary liver cancer characterized by mutation inactivation of NBS1, a DNA repair associated gene.     

Paralog-divergent Features May Help Reduce Off-target Effects of Drugs: Hints from Glucagon Subfamily Analysis

Side effects from targeted drugs remain a serious concern. One reason is the nonselective binding of a drug to unintended proteins such as its paralogs, which are highly homologous in sequences and have similar structures and drug-binding pockets. To identify targetable differences between paralogs, we analyzed two types(type-I and type-II) of functional divergence between two paralogs in the known target protein receptor family G-protein coupled receptors(GPCRs) at the amino acid level. Paralogous protein receptors in glucagon-like subfamily, glucagon receptor(GCGR) and glucagon-like peptide-1 receptor(GLP-1R), exhibit divergence in ligands and are clinically validated drug targets for type 2 diabetes. Our data showed that type-II amino acids were significantly enriched in the binding sites of antagonist MK-0893 to GCGR, which had a radical shift in physicochemical properties between GCGR and GLP-1R. We also examined the role of type-I amino acids between GCGR and GLP-1R. The divergent features between GCGR and GLP-1R paralogs may be helpful in their discrimination, thus enabling the identification of binding sites to reduce undesirable side effects and increase the target specificity of drugs.     

超声造影引导下行微波消融术治疗肝癌的临床效果分析

目的：评估超声造影（CEUS）gl导下微波消融（MwA）治疗肝癌的有效性及应用价值。方法：108例肝癌患者,共147个病灶经MWA治疗。根据患者在做常规超声检查时,是否有扫查不清晰的结节或其他影像学检查提示可能存在多发结节等,分成CEUS族和对照组。CEUS组41名患者,年龄（57．9±7．8）岁,共57个病灶,平均直径（2．4±1．5）cm,经超声造影引导下行微波消融治疗。对照组67名患者,年龄（55．5±8．9）岁,共90个病灶,平均直径（2．6±1．7）cm,常规超声引导下同种条件行微波消融治疗。治疗后对两组患者进行6～12个月随访。结果：CEUS组的57个病灶均清晰显示其位置、数目、大小、边界、形态,完全消融的结节为55个,未完全消融的结节为2个。对照组67名患者的90个病灶,完全消融的病灶78个;未完全消融的结节12个。CEUS组完全消融率高于病例对照组（96．49％VS86．67％;P〈0．05）。随访6～12个月后发现,CEUS组完全消融率高于病例对照组（P〈0．05）,差异有统计学意义。结论：CEUS能更好的显示病灶的位置、数目、大小,更精确显示病灶边界、范围,造影引导下MWA是一种有效提高完全消融效率的方法,具有较大应用价值。     

脾切除术对肝炎后肝硬化患者肝癌发生的影响

目的:观察脾切断流术对肝硬化门静脉高压症患者肝功的影响.方法:回顾分析2010.4-2011.12在我科住院的肝硬化门静脉高压症患者行脾切除术前后肝功能的变化.应用超声多普勒分别检测手术前后门静脉血流量(PVF)、肝动脉血流量(HAF),术中水柱法测量自由门静脉压力(FPP);回顾分析(通过对病历查阅获取)上述患者Child分级指标以及肝功能指标.用SAS 8.0对获取的指标采用配对资料t检验和卡方检验.结果:肝硬化门静脉高压症患者共480例,行脾脏切除加贲门周围血管离断术450例,单纯脾切除30例.PVF由术前的(1368± 402)ml/min减少至术后的(986± 295)ml/min(P＜0.01);HAF由术前的(412± 68)ml/min增加至术后的(526± 132)ml/min(P＜0.01);FPP由术前的(30± 6)cm H2O降至术后的(26± 5)cm H2O(P＜0.01).R15 ICG由术前的(18±8)％降至术后的(16±7)％(P＞0.05).肝功能Child分级A级患者比率由术前85％增加至术后95％(P＜0.01).其它肝脏功能指标如总胆红素、凝血酶原延长时间以及腹水消失率都明显好转.结论:脾切断流术后门静脉血流量虽然减少,但是肝动脉血流量增加及术后门体分流减少均有利于肝功能恢复.     

Generation of Subcutaneous and Intrahepatic Human Hepatocellular Carcinoma Xenografts in Immunodeficient Mice

In vivo experimental models of hepatocellular carcinoma (HCC) that recapitulate the human disease provide a valuable platform for research into disease pathophysiology and for the preclinical evaluation of novel therapies. We present a variety of methods to generate subcutaneous or orthotopic human HCC xenografts in immunodeficient mice that could be utilized in a variety of research applications. With a focus on the use of primary tumor tissue from patients undergoing surgical resection as a starting point, we describe the preparation of cell suspensions or tumor fragments for xenografting. We describe specific techniques to xenograft these tissues i) subcutaneously; or ii) intrahepatically, either by direct implantation of tumor cells or fragments into the liver, or indirectly by injection of cells into the mouse spleen. We also describe the use of partial resection of the native mouse liver at the time of xenografting as a strategy to induce a state of active liver regeneration in the recipient mouse that may facilitate the intrahepatic engraftment of primary human tumor cells. The expected results of these techniques are illustrated. The protocols described have been validated using primary human HCC samples and xenografts, which typically perform less robustly than the well-established human HCC cell lines that are widely used and frequently cited in the literature. In comparison with cell lines, we discuss factors which may contribute to the relatively low chance of primary HCC engraftment in xenotransplantation models and comment on technical issues that may influence the kinetics of xenograft growth. We also suggest methods that should be applied to ensure that xenografts obtained accurately resemble parent HCC tissues.     

超声造影引导下行微波消融术治疗肝癌的临床效果分析*

摘要目的：评估超声造影(CEUS)引导下微波消融(MWA)治疗肝癌的有效性及应用价值。方法：108 例肝癌患者,共147个病灶经 MWA治疗。根据患者在做常规超声检查时,是否有扫查不清晰的结节或其他影像学检查提示可能存在多发结节等,分成CEUS 族和对照组。CEUS组41 名患者,年龄(57.9± 7.8）岁,共57 个病灶,平均直径（2.4± 1.5）cm,经超声造影引导下行微波消融治疗。 对照组67 名患者,年龄（55.5± 8.9）岁,共90 个病灶,平均直径（2.6± 1.7）cm,常规超声引导下同种条件行微波消融治疗。治疗后 对两组患者进行6～12 个月随访。结果：CEUS组的57 个病灶均清晰显示其位置、数目、大小、边界、形态,完全消融的结节为55 个,未完全消融的结节为2个。对照组67 名患者的90 个病灶,完全消融的病灶78 个;未完全消融的结节12 个。CEUS组完全消 融率高于病例对照组(96.49 %VS 86.67 %;P< 0.05）。随访6～12 个月后发现,CEUS 组完全消融率高于病例对照组(P< 0.05）,差 异有统计学意义。结论：CEUS能更好的显示病灶的位置、数目、大小,更精确显示病灶边界、范围,造影引导下MWA 是一种有效 提高完全消融效率的方法,具有较大应用价值。     

Plasma Protein Biomarkers of Hepatocellular Carcinoma in HCV-Infected Alcoholic Patients with Cirrhosis

Hepatocellular carcinoma (HCC) is one of the most common and lethal cancers in the world, with limited options for treatment unless timely diagnosed. Chronic hepatitis C virus (HCV) infection and persistent heavy alcohol consumption are independent risk factors for HCC development, which may induce a specific protein expression pattern different from those caused separately. The aim of the study was to identify protein biomarkers for the detection of HCC in HCV-infected alcoholic patients with cirrhosis in order to improve survival. We compared protein expression profiles of plasma samples from 52 HCV-infected alcoholic patients with and without HCC, using 2-D DIGE coupled with MALDI-TOF/TOF mass spectrometry. The 2-D DIGE results were analyzed statistically using Decyder software, and verified by western-blot and ELISA. In plasma samples from HCV-infected alcoholic patients, we found significantly differential expression profiles of carboxypeptidase-N, ceruloplasmin (CP), complement component 4a (C4a), fibrinogen-alpha (FGA), immunoglobulin mu chain C region, serum albumin, and serum paraoxonase/arylesterase 1 (PON1). Deregulation of plasma/serum levels of the identified proteins was associated to HCV, ethanol consumption, and/or HCC progression. In the validation through ELISA, C4a serum concentration was increased in HCC patients (2.4±1 ng/mg vs 1.8±0.6 ng/mg; p = 0.029), being the only independent predictor of HCC in the multivariate analysis (OR = 2.15; p = 0.015), with an AUROC = 0.70. The combination of C4a, FGA, CP and PON1 improved slightly the predictive ability of C4a alone (AUROC 0.81). In conclusion, we identified proteins related to acute-phase response, oxidative stress, or immune response, whose differential expression in plasma may be attributed to the presence of HCC. Among them, C4a, and its combination with CP, FGA and PON1, could be considered as potentially reliable biomarkers for the detection of HCC in HCV-infected alcoholic patients.     

Smo 蛋白在肝癌和肝硬化中的表达及临床意义

目的:研究Smoothened(SMO)在肝癌和肝硬化中的表达及临床意义。方法:选取组织标本后,运用石蜡包埋,切片后,HE染色,构建组织芯片,免疫组织化学和原位杂交方法检测Smo蛋白在肝癌和肝硬化中的表达。结果:Smo蛋白在肝癌细胞浆、良性肝肿瘤组织细胞浆、肝硬化组织中强染色,在正常组织中无染色。并且在典型肝硬化中强染色,在中度肝硬化中弱阳性。结论:Smo蛋白表达与肝癌的发生有关,Smo基因的高表达可能激活某种机制而参与诱导肝癌的发生。可能是通过异常激活Sonic hedgchog信号通路,从而诱导肝癌的发生与发展。     

HBV相关肝癌组织中微生物宏蛋白质组特征分析

越来越多的证据表明微生物菌群和肝脏疾病有着密切联系,但是HBV相关肝细胞癌(hepatitis B virus(HBV)-related hepatocellular carcinoma,HCC)人群肝脏组织内的微生物菌群种类和数量仍不清楚.本研究对来自165个HCC人群的肝癌组织和对应癌旁组织的质谱数据进行宏蛋白质组...     

Interferon-Based Therapy Decreases Risks of Hepatocellular Carcinoma and Complications of Cirrhosis in Chronic Hepatitis C Patients

Background

Interferon-based therapy (IBT) has been the standard of care for hepatitis C virus (HCV) infection. However, conflicting results exist regarding the effects of IBT on risk of developing hepatocellular carcinoma (HCC) and cirrhosis-associated complications, and most included highly selected patients.

Methods

This 8-year cohort study was based on the Longitudinal Health Insurance Database 2000 (LHID 2000) consisting of 1,000,000 beneficiaries randomly selected from all Taiwan National Health Insurance enrollees in 2000 (>23.7 million). Patients with newly detected HCV infections (n = 11,264) were classified based on treatment and clinical outcomes. IBTs were defined as regimens that included interferon- alfa, pegylated interferon- alfa -2a, or pegylated interferon- alfa -2b for at least 3 months. The Cox proportional hazards models were used to estimate the hazard ratio (HR) and associated confidence interval (CI) of HCC and cirrhosis-associated complications for IBT.

Results

The 8-year incidence rate for HCC was 3.9% among patients who received IBT and 5.6% among those who did not. The HCC-free survival rate was significantly higher among patients receiving IBT during the 8-year period than their counterpart (adjusted HR, 0.50; 95% CI, 0.31–0.81; P = .004). Similarly, the event-free survival rates for esophageal variceal bleeding (adjusted HR, 0.45; 95% CI, 0.22–0.91; P = .026), hepatic encephalopathy (adjusted HR, 0.38; 95% CI, 0.21–0.69; P = .001), ascites (adjusted HR, 0.28; 95% CI, 0.14–0.57; P<.001), and cirrhosis (adjusted HR, 0.63; 95% CI, 0.44–0.91; P = .013) were significantly higher among patients who received IBT than those who did not, after adjustment for associated factors.

Conclusion

Treatment with interferon may reduce the 8-year risk of HCC and cirrhosis-associated complications in patients with chronic HCV infection.     

Relationship between Enhanced Intensity of Contrast Enhanced Ultrasound and Microvessel Density of Aortic Atherosclerostic Plaque in Rabbit Model

The aim of this study was to evaluate the relationship between enhanced intensity of contrast enhanced ultrasound and microvessel density of aortic atherosclerotic plaque in rabbit model. The abdominal aortas of thirty-six male New Zealand rabbits were damaged by balloon expansion and the animals were then fed a high fat diet for 12 weeks. Twenty-seven plaques on the near aortic wall were detected using conventional ultrasound examination. The maximum thickness of each plaque was recorded. CEUS was performed on these 27 plaques and the time-intensity curves (TICs) were analyzed offline. Using the quantitative ACQ software, features such as the arrival time (AT), time to peak (TTP), baseline intensity (BI), peak intensity (PI) and enhanced intensity (EI) (EI = PI-BI) were assessed. Inter- and intra-observer agreement of EI were assessed using the Bland-Altman test. After CEUS examination, the rabbits were sacrificed for pathological examination and CD34 monoclonal antibody immunohistochemical detection. Microvessel density (MVD) was counted under the microscope. The relationship between indexes of CEUS and the level of MVD was analyzed. There was a good positive linear correlation between EI and MVD (γ = 0. 854, P<0. 001), the intraclass correlations for inter- and intra-observer agreement for EI were 0.73 and 0.82 respectively, suggesting that EI may be act as a useful index for plaque risk stratification in animal models.     

Ultrasound Exposure Improves the Targeted Therapy Effects of Galactosylated Docetaxel Nanoparticles on Hepatocellular Carcinoma Xenografts

Purpose

The distribution of targeted nanoparticles in tumor tissue is affected by a combination of various factors such as the physicochemical properties of the nanoparticles, tumor hemoperfusion and tumor vascular permeability. In this study, the impact of the biological effects of ultrasound on nanoparticle targeting to liver carcinoma was explored.

Methods

The copolymer MePEG-PLGA was used to prepare the galactosylated docetaxel nanoparticles (GDN), and the physical and chemical properties as well as the acute toxicity were then assayed. The impact of ultrasound exposure (UE) on tumor hemoperfusion was observed by contrast-enhanced ultrasonography (CEUS), and the distribution of docetaxel in tumors and liver were detected by high performance liquid chromatography (HPLC). In the GDN combined with UE treatment group, the mice were injected intravenously with GDN, followed by ultrasound exposure on the human hepatocellular carcinoma xenografts. Twenty-eight days post-administration, the tumor growth inhibition rate was calculated, and the expression of Survivin and Ki67 in tumor tissues were determined by immunohistochemistry assay and quantitative real-time PCR.

Results

The mean size of prepared liver-targeting nanoparticles GDN was 209.3 nm, and the encapsulation efficiency was 72.28%. The median lethal dose of GDN was detected as 219.5 mg/kg which was about four times higher than that of docetaxel. After ultrasound exposure, the tumor peak - base intensity difference value, examined by CEUS, increased significantly. The drug content in the tumor was 1.96 times higher than in the GDN treated control. In vivo, GDN intravenous injection combined with ultrasound exposure therapy achieved the best anti-tumor effect with a tumor growth inhibition rate of 74.2%, and the expression of Survivin and Ki67 were significantly decreased as well.

Conclusion

Ultrasound exposure can improve targeting nanoparticles accumulation in the tumor, and achieve a synergism antitumor effect on the hepatocellular carcinoma xenografts.     

基于生物信息学分析的肝癌核心基因的筛选及验证

本研究基于GEO数据库,选取由慢性乙型肝炎诱导的肝细胞癌芯片数据GSE121248为研究对象,利用GEO2R软件分析数据,筛选出差异表达基因,利用DAVID数据库进行GO分析和KEGG pathway富集分析.利用STRING数据库构建PPI网络,分析筛选核心基因.利用GEPIA对核心基因的表达进行验证,Kaplan ...     

CD90在肝细胞肝癌组织中的表达及其与肝癌临床病理特征的关系

目的:探讨CD90在肝细胞肝癌(HCC)患者肿瘤组织中的表达及其与临床病理参数的关系,为HCC的临床治疗提供参考。方法:选择2014年3月-2015年3月在我院接受治疗的HCC患者80例,收集患者肿瘤组织及癌旁组织,另选择同期就诊的80例正常肝脏组织作为对照组。观察并比较CD90在肝癌组织、癌旁组织及正常肝脏组织中的表达情况。结果:CD90在肝癌组织、癌旁组织及正常肝脏组织中呈不同程度阳性表达(P0.05)。CD90在肝癌组织和癌旁组织中的表达显著高于正常肝脏组织,差异具有统计学意义(P0.05)。CD90在肝癌组织中的表达显著高于癌旁组织,差异具有统计学意义(P0.05)。CD90在HCC组织中的表达与肿瘤大小、肝内转移、TMN分期和病理分级有关(P0.05)。结论:CD90在肝癌组织中呈高表达,其表达水平与肝癌临床病理参数有关,可以作为HCC诊断、治疗及预后评估的参考指标。     

Contrast Enhanced Ultrasound Imaging for Assessment of Spinal Cord Blood Flow in Experimental Spinal Cord Injury

Reduced spinal cord blood flow (SCBF) (i.e., ischemia) plays a key role in traumatic spinal cord injury (SCI) pathophysiology and is accordingly an important target for neuroprotective therapies. Although several techniques have been described to assess SCBF, they all have significant limitations. To overcome the latter, we propose the use of real-time contrast enhanced ultrasound imaging (CEU). Here we describe the application of this technique in a rat contusion model of SCI. A jugular catheter is first implanted for the repeated injection of contrast agent, a sodium chloride solution of sulphur hexafluoride encapsulated microbubbles. The spine is then stabilized with a custom-made 3D-frame and the spinal cord dura mater is exposed by a laminectomy at ThIX-ThXII. The ultrasound probe is then positioned at the posterior aspect of the dura mater (coated with ultrasound gel). To assess baseline SCBF, a single intravenous injection (400 µl) of contrast agent is applied to record its passage through the intact spinal cord microvasculature. A weight-drop device is subsequently used to generate a reproducible experimental contusion model of SCI. Contrast agent is re-injected 15 min following the injury to assess post-SCI SCBF changes. CEU allows for real time and in-vivo assessment of SCBF changes following SCI. In the uninjured animal, ultrasound imaging showed uneven blood flow along the intact spinal cord. Furthermore, 15 min post-SCI, there was critical ischemia at the level of the epicenter while SCBF remained preserved in the more remote intact areas. In the regions adjacent to the epicenter (both rostral and caudal), SCBF was significantly reduced. This corresponds to the previously described “ischemic penumbra zone”. This tool is of major interest for assessing the effects of therapies aimed at limiting ischemia and the resulting tissue necrosis subsequent to SCI.