Vitamin D-binding protein synthesized by a human hepatocellular carcinoma cell line

The binding protein for 25-hydroxycholecalciferol was studied in the medium spent for the culture of HuH-7 cells, which were originally derived from a human hepatocellular carcinoma tissue. The binding protein for 25-hydroxycholecalciferol synthesized by HuH-7 cells was immunologically similar to vitamin D-binding protein in human serum and had an inter-alpha mobility. A sedimentation coefficient of 4.1 S was found on sucrose density gradient analyses. The molecular weight was estimated to be approximately 58,000 by gel filtration on a standardized column of Sephadex G-150. When mixed with filamentous actin purified from rabbit skeletal muscle, it depolymerized filamentous actin and bound to monomeric, globular actin to make a 5.5 S 1:1 molar complex with a molecular weight of approximately 100,000. These results support the conclusion that HuH-7 cells produce a functional vitamin D-binding protein.     

血清肿瘤标志物的联合检测在原发性肝癌中的诊断价值

目的探讨血清甲胎蛋白(AFP)、癌胚抗原(CEA)、癌抗原125(CA125)、CA199的联合检测在原发性肝癌(PHC)中的诊断价值。方法选取2013年7月至2014年6月收治的67例PHC患者(PHC组)、83例良性肝病患者(良性肝病组),另选取同期125例体检健康者(对照组),采用化学发光法分别检测所有研究对象的血清AFP、CEA、CA125、CA199水平并进行统计学分析。结果 PHC组血清AFP、CEA、CA125和CA199阳性率分别为71.6%、40.3%、53.7%、58.2%,均高于良性肝病组与对照组,比较差异均有统计学意义(P0.05);4项肿瘤标志物联合检测在PHC组的阳性率为91.0%,高于各肿瘤标志物单项检测的阳性率,比较差异均有统计学意义(P0.05)。结论 AFP、CEA、CA125、CA199联合检测能够明显提高PHC的诊断阳性率,有助于PHC的早期诊断。     

Contrast-enhanced ultrasonography for blood flow detection in hepatocellular carcinoma during lenvatinib therapy

Purpose

Blood flow reduction after initiation of lenvatinib therapy may not always indicate tumor necrosis. This study aimed to compare the blood flow detectability of contrast-enhanced ultrasonography (CEUS), contrast-enhanced computed tomography (CT), and contrast-enhanced magnetic resonance imaging (MRI) in hepatocellular carcinoma (HCC) during lenvatinib therapy.

Methods

A total of 12 cases underwent CEUS and contrast-enhanced CT/MRI within 2 weeks during lenvatinib therapy. Vascularity on CEUS and CT/MRI was compared.

Results

At the time of CEUS examination, the median period from the start of lenvatinib was 227?±?210 (31–570) days. CEUS showed hyperenhancement in eight cases (66.7%), hypoenhancement in two cases (16.7%), and no enhancement in one case (8.3%), while CT/MRI showed hyperenhancement in one case (8.3%), ring enhancement in three cases (25.0%), and hypoenhancement in eight cases (66.7%) (p?=?0.007). Transarterial chemoembolization (n?=?3), radiofrequency ablation (n?=?2), and stereotactic body radiation therapy (n?=?2) were performed after blood flow detection by CEUS.

Conclusions

The viability of the HCC should be confirmed using CEUS when contrast-enhanced CT/MRI reveals lesion hypoenhancement during lenvatinib therapy.

     

血清 ５ 项肿瘤标志物检测对肝癌的临床应用价值

：目的 探讨血清甲胎蛋白（ＡＦＰ）、甲胎蛋白异质体（ＡＦＰ Ｌ３）、甲胎蛋白异质体比率（ＡＦＰ Ｌ３％）、异常凝血酶原（ＤＣＰ）、 高尔基体蛋白 ７３（ＧＰ７３）单项及联合检测在慢性肝病、肝硬化和肝癌中的临床应用价值。方法 回顾性分析 ２０１９ 年 １０ 月至 ２０２０ 年 １２ 月吉林大学第一医院就诊的 ６ ２０２ 例住院及门诊患者。采用磁微粒化学发光免疫分析法检测各患者上述 ５ 项指 标。分别将肝癌治疗前及治疗后未缓解组、肝癌治疗后缓解组与慢性肝病及肝硬化组检测结果进行两独立样本的秩和检验 分析。通过 ＲＯＣ 曲线及 Ｌｏｇｉｓｔｉｃ 回归评估 ５ 项指标单项及联合检测在肝癌治疗前及治疗后未缓解组中的诊断效能及关联程 度。对 ５ 项指标单项及联合检验阳性率以及诊断肝癌的方法学进行评价。结果 肝癌治疗前及治疗后未缓解组、肝癌治疗后 缓解组该 ５ 项指标与慢性肝病及肝硬化组比较的差异均有统计学意义（Ｐ均＜０．０５）。５ 项指标诊断肝癌治疗前及治疗后未缓 解组的 ＲＯＣ 曲线下面积（ＡＵＣＲＯＣ）为 ０．８００ ～ ０．９１４，ｃｕｔ ｏｆｆ 值分别为：ＡＦＰ ７．７２ ｎｇ ／ ｍＬ，ＡＦＰ Ｌ３ ０．６０ ｎｇ ／ ｍＬ，ＡＦＰ Ｌ３％ ５．０％， ＤＣＰ ３６．７９ ｎｇ ／ ｍＬ，ＧＰ７３ ７８．９７ ｎｇ ／ ｍＬ；敏感性 ６９．０４％ ～ ８２．９４％，特异性 ７８．３４％ ～ ９８．２２％，约登指数 ０．４７６～ ０．８１２。联合检测诊 断肝癌治疗前及治疗后未缓解组的 ＡＵＣＲＯＣ为 ０．９２８～ ０．９９６，敏感性 ８３．２０％ ～９７．６７％，特异性 ９４．８９％ ～９８．１２％，约登指数 ０．８１１～ ０．９５５。单项检测阳性率分别为：慢性肝病及肝硬化组 ０． ６３％（ＡＦＰ Ｌ３％）～ ６． ３７％（ＡＦＰ），肝癌治疗前及治疗后未缓解组 ３２．０６％（ＧＰ７３）～ ７９．７７％（ＡＦＰ），肝癌治疗后缓解组 ５．７７％（ＡＦＰ Ｌ３％）～ １７．６３％（ＤＣＰ）；联合检验阳性率分别为：慢性肝病及 肝硬化组 ３．６７％（ＡＦＰ Ｌ３％＋ＧＰ７３）～ １１．８２％（５ 项联合），肝癌治疗前及治疗后未缓解组 ８５．８０％（ＡＦＰ＋ＡＦＰ Ｌ３％）～ ９９．８２％（５ 项联合），肝癌治疗后缓解组 １２．６７％（ＡＦＰ ＋ＡＦＰ Ｌ３％）～ ３２．９４％（５ 项联合）。诊断肝癌的敏感性分别为 ２２．６２％（ＧＰ７３）～ ６９．７７％（５ 项联合），特异性分别为 ８８．１８％（５ 项联合）～ ９９．３７％（ＡＦＰ Ｌ３％），总正确率 ４７．４４％（ＧＰ７３）～ ７５．９３％（５ 项联合），阳 性预测值 ９２．１６％（５ 项联合）～ ９９．３３％（ＡＦＰ Ｌ３％），阴性预测值 ３８．５９％（ＧＰ７３）～ ５９．４３％（５ 项联合）。结论 慢性肝病、肝硬 化及肝癌治疗前后 ５项血清学指标发生不同程度改变，联合检测对原发性肝癌的辅助诊断和评价疗效具有重要的临床应用价值。     

肝细胞癌分子标记物及相关基因的研究进展

肝细胞肝癌是世界范围内常见的恶性肿瘤之一,其在分化形成过程中可出现的一些异常蛋白表达(如AFP、CD34和GPC3等),有助于肝细胞癌的诊断与鉴别诊断.在肝细胞癌发生、发展机制的研究中,发现有众多的癌基因和抑癌基因的突变或表达异常参与肿瘤的形成和分化,最新研究还发现非编码蛋白microRNA亦参与了肝细胞癌的形成.     

Recent advances in tumor markers of human hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is one of the most devastating malignancies in the world and is the third most common cause of cancer-related death in Korea. Because most HCC are accompanied by chronic liver disease that results from hepatitis B or C viruses, prognosis is still poor even after surgical resection of the tumor. Moreover, diagnosis of advanced HCC still leads to an extremely bleak prognosis. Earlier detection of HCC, therefore, could improve patient survival. Accordingly, the development of tumor markers that can detect HCC at even earlier stages is essential. The functions of tumor markers include prediction of prognosis or therapeutic response as well as diagnosis or screening of cancer. Possible candidate tumor markers may be quantitative alterations in DNA-, RNA- or protein-based molecules in tumorous conditions assessed by various technologies, e.g. serological assays, microarrays, mass spectrometry and proteomics. However, validation and clinical implementation is needed after the discovery of novel genes. An ideal tumor marker for HCC would be sensitive and specific enabling to differentiate it at an early stage from premalignant lesions like dysplastic nodules. In addition, the marker should be easily measurable, reproducible and minimally invasive. Although it is important to identify new biomarkers for HCC, the validation and cost-effectiveness of those markers as diagnostic or prognostic tools need confirmation in large-scale studies in clinical practice.     

Obstructive jaundice caused by hepatocellular carcinoma: detection by endoscopic sonography.

Tumor thrombus in the extrahepatic biliary tree is a rare mechanism of obstructive jaundice. We present a patient with a minute hepatocellular carcinoma in the caudate lobe that invaded the common hepatic duct and caused biliary obstruction. Endoscopic sonography showed a tumor thrombus with central echogenicity and a "nodule-in-nodule" pattern and suggested the correct diagnosis.     

Candidate molecular markers for histological diagnosis of early hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is one of the most common malignant tumors. HCC occurs mainly in chronically diseased livers, e.g. following hepatitis B and C infection. These high-risk patients are closely followed up, and increasing numbers of small equivocal lesions are detected by imaging diagnosis. They are now widely recognized as precursor or early-stage HCCs and are classified as dysplastic nodule or early HCC. These lesions lack typical imaging and histology of ordinary HCC and do not show elevated serum markers of alpha-fetoprotein and PIVKA-II, for example. Molecular analysis of these lesions would help to develop molecular markers for objective histological diagnosis of early HCC and possibly new serum markers for early detection of HCC. It has been reported that HSP70, CAP2, glypican 3 and glutamine synthetase could serve as molecular markers for early HCC. Further analysis is expected to evaluate their usefulness in routine pathological diagnosis including biopsy diagnosis and also as serum markers for early detection of HCC.     

原发性肝癌的分子诊断标记物研究进展

在我国,原发性肝癌具有较高的发病率和死亡率,严重威胁我国人民的生命和健康.对肝癌高危人群进行筛查,早期发现、早期诊断、早期治疗,是提高肝癌疗效的关键.肝癌特异性的血清分子诊断标记物是当前肝癌早期筛查的重要手段,已广泛应用于临床工作中.随着新的研究技术的不断应用和肝肿瘤发生机制认识的不断深入,有越来越多的原发性肝癌相关分...     

Serum F protein: a new sensitive and specific test of hepatocellular damage

F protein is a 44-kDa protein which is found mostly in the liver and circulates at much lower concentrations in the serum. Serum F protein has been measured, using a recently developed radioimmunoassay, in a variety of diseases and was appreciably raised only in patients with hepatocellular damage. The serum F protein concentration was a more sensitive and specific marker of liver damage than conventional liver function tests and showed a close correlation with the histological assessment of liver damage (r = 0.86, P less than 0.001). This new marker may be of value in the diagnosis and treatment of liver disease.     

Genetic and epigenetic silencing of SCARA5 may contribute to human hepatocellular carcinoma by activating FAK signaling

The epigenetic silencing of tumor suppressor genes is a crucial event during carcinogenesis and metastasis. Here, in a human genome-wide survey, we identified scavenger receptor class A, member 5 (SCARA5) as a candidate tumor suppressor gene located on chromosome 8p. We found that SCARA5 expression was frequently downregulated as a result of promoter hypermethylation and allelic imbalance and was associated with vascular invasion in human hepatocellular carcinoma (HCC). Furthermore, SCARA5 knockdown via RNAi markedly enhanced HCC cell growth in vitro, colony formation in soft agar, and invasiveness, tumorigenicity, and lung metastasis in vivo. By contrast, SCARA5 overexpression suppressed these malignant behaviors. Interestingly, SCARA5 was found to physically associate with focal adhesion kinase (FAK) and inhibit the tyrosine phosphorylation cascade of the FAK-Src-Cas signaling pathway. Conversely, silencing SCARA5 stimulated the signaling pathway via increased phosphorylation of certain tyrosine residues of FAK, Src, and p130Cas; it was also associated with activation of MMP9, a tumor metastasis–associated enzyme. Taken together, these data suggest that the plasma membrane protein SCARA5 can contribute to HCC tumorigenesis and metastasis via activation of the FAK signaling pathway.     

Development of a simple whole blood panel test for detection of human heart-type fatty acid-binding protein.

OBJECTIVE: For the diagnosis of acute myocardial infarction (AMI), we have developed a rapid and simple whole blood panel test for the detection of human heart-type fatty acid-binding protein (H-FABP) using one-step immunochromatography. METHODS AND RESULTS: We have developed a whole blood panel test for rapid detection of human H-FABP using a one-step immunochromatography technique. The result of this panel test was not affected by the other contents of the blood such as bilirubin, hemoglobin and others. Furthermore, no cross-reactivity of the antibodies was found with other cardiac markers or other tissue-type FABPs. The result of this panel test was similar to the diagnostic cut-off value, 6.2 ng of H-FABP per mL of serum which was evaluated by the enzyme-linked immunosorbent assay (ELISA). CONCLUSION: We have developed a simple one-step immunochromatography technique to detect H-FABP in whole blood sample. Further studies are required to identify the value of this point-of-care testing (POCT) as a diagnostic marker for AMI.     

4种标记蛋白抗体测定在鼻咽癌体检筛查及诊断中的应用

目的检测EB病毒Rta-IgG、病毒壳抗原(VCA-IgA)、Zta-IgA、核抗原(NA1-IgA)抗体在鼻咽癌血清中的表达水平,以及应用于体检中的最适指标。方法采用酶联免疫吸附试验(ELISA)分别检测健康体检者和鼻咽癌患者血清中的Rta-IgG、VCA-IgA、ZTA-IgA、NA1-IgA抗体水平,比较4项指标在鼻咽癌患者中的阳性预测值和健康体检者中的表达水平及阴性预测值。结果 20例确诊为鼻咽癌的患者检测结果数据可见,VCA-IgA阳性预测值为70%,NA1-IgA、Rta-IgG阳性预测值为85%,Zta-IgA阳性预测值为90%;对1 000例健康体检者4种标记蛋白抗体检测结果数据分析可见,Zta-IgA阴性预测值为83.3%,VCA-IgA阴性预测值为89.5%,Rta-IgG阴性预测值为91.9%,NA1-IgA阴性预测值为91.5%。结论在ELISA检测EB病毒4种标记蛋白抗体中,如果单独采用一种,则以Rta-IgG或NA1-IgA为佳,比较适合于健康体检人群的鼻咽癌体检筛查工作。如采用NA1-IgA、Zta-IgA、VCA-IgA或Zta-IgA、VCA-IgA、Rta-IgG 3项联合检测,则可提高阳性检出率。     

Molecular evaluation of apoptotic versus antiapoptotic angiogenic markers in hepatocellular carcinoma

Objective

To assess the role of HO-1 in HCC progression and to study the expression of apoptotic factors represented by TNF-α, and Fas-L versus antiapoptotic and angiogenic factors represented by HO-1, TGF-β, HGF, and VEGF in HCC compared to non cancerous cirrhotic liver.

Design and methods

Liver biopsies were taken from twelve patients with grade II HCC confined to the liver and twelve patients with non cancerous liver cirrhosis (served as control). RT-PCR of previous genes was evaluated.

Results

HO-1, VEGF, HGF, and TNF-α genes were significantly increased (P < 0.05) in HCC compared to control. Fas-L showed a significant decrease (P < 0.05) in HCC compared to control. TGF-β was higher in HCC than control but the difference was not statistically significant (P > 0.05). HGF showed significant positive correlation with HO-1 (r = 0.8217, P = 0.001).

Conclusion

HCC is associated with increased expression of VEGF, HGF, and TGF-β, and with suppression of Fas-L. In addition, HO-1 is highly significantly expressed in HCC. The significant positive correlation between HO-1 and HGF was first reported in Egyptian human liver biopsies, and this suggests that it may play a role in the progression of hepatocellular carcinoma.     

肝癌侧支供血的临床研究

目的：探讨肝癌侧支供血对肝癌化疗栓塞（ＴＡＥ）治疗效果的影响。材料和方法：通过１９２例肝癌腹腔动脉造影表现进行分析。结果：显示（ＴＡＥ）疗效与肿瘤物供血情况有关,单一动脉供血,无侧支供血者疗效较好,有侧支供血者疗效差。结论：侧支供血浊影响疗效的重要因素,指出ＴＡＥ时对侧支供血动脉应作永久性栓塞。     

Dysregulated expression of dickkopfs for potential detection of hepatocellular carcinoma

The prognosis for hepatocellular carcinoma (HCC) remains dismal due to the lack of diagnostic markers for early detection. This review will discuss the clinical potential of the dickkopf (DKK) family members as diagnostic and/or prognostic markers for HCC. In comparison to serum α-fetoprotein (AFP) level, which remains the gold standard for HCC diagnosis, high serum DKK1 levels have higher diagnostic value for HCC, especially for AFP-negative HCC, and can distinguish HCC from non-malignant chronic liver diseases. Additionally, the combination of serum DKK1 and AFP levels enhances diagnostic accuracy for HCC compared to serum DKK1 or AFP levels alone. Although DKK1 offers potential for its use in HCC diagnosis this review will discuss the challenges facing DKK1 and also shed some light on recent developments on the remaining DKK family members: DKK2, DKK3 and DKK4.     

血清MMP9蛋白ELISA检测方法的建立及在肝癌患者血清检测中的初步应用

目的建立基质金属蛋白酶9（MMP9）的双抗体夹心ELISA检测方法，探讨血清中MMP9在正常人及肝癌患者中的差异。方法从人肝组织中经过反转录扩增出MMP9基因721—1156bp区段，插入到原核表达质粒pQE30中，在大肠埃希菌M15中诱导蛋白表达。以纯化的融合蛋白HIS—MMP9为抗原免疫实验用兔及豚鼠，得到的MMP9抗血清经过纯化后，建立双抗体夹心ELISA法。对227份正常及193份肝癌患者血清中的MMP9进行检测。结果SDS—PAGE显示所表达的HIS—MMP9融合蛋白相对分子质量约为17000；以原核表达蛋白为抗原制备的MMP9抗血清可有效地识别重组MMP9和中性粒细胞和HepG2细胞内源性MMP9蛋白；利用建立的MMP9 ELISA检测技术发现肝细胞癌患者血清中MMP9的含量高于正常人群，差异具有统计学意义，P〈0．001。结论建立的双抗体夹心ELISA方法可用于血清MMP9的检测，为建立以MMP9为检测指标的肝细胞癌患者转移复发筛查方法提供了科学基础。     

CD133、CD44和PSCA在肝癌组织中的表达及其意义

目的 研究CD133、CD44和PSCA在肝癌组织、癌旁肝组织和肝癌细胞系中表达的特异性.方法 采用免疫组化SP法检测CD133、CD44和PSCA在70例肝细胞癌(HCC)、50例肝内胆管细胞癌(ICC)组织及培养的HepG2细胞的表达.结果 CD133、CD44和PSCA在HCC组织中,阳性率分别为58.6％ (41/70)、61.4％ (43/70)和40％ (28/70);在ICC组织中的阳性率分别为56％ (28/50)、52％(26/50)和44％(22/50);统计学分析显示三者在HCC和ICC的阳性率差异不显著(x2=0.824,P＞0.05);但PSCA在ICC的阳性表达强度明显高于HCC(P＜0.05).CD133、CD44、PSCA阳性细胞在HCC、ICC和HepG2培养细胞中的分布均存在散在和成簇/片状两种形式,其中CD133以散在分布为主,而CD44和PSCA则多呈成簇/片状分布.在HCC和ICC癌的旁组织中,CD133、CD44和PSCA阳性表达率分别为51.4％(36/70)、47.1％(33/70)和32.9％(23/70)以及46％(23/50)、32％(16/50)和36％(18/50),比较3种蛋白在HCC和ICC的癌旁组织中的阳性率无明显差异(P＞0.05).CD133、CD44、PSCA在HCC、ICC及其癌旁组织中的阳性表达有明显的一致性.结论 CD133、CD44和PSCA可能均是肝癌干细胞的特异性表面标记物,其中CD133的特异性最强;而PSCA可能是ICC较为特异的癌干细胞表面标记物.     

原发性肝癌的新型肿瘤标志物:甲胎蛋白异质体AFP-L3、高尔基体蛋白GP73

原发性肝癌(hepatocellular carcinoma,HCC)是严重威胁我国人民健康的恶性肿瘤之一,近年来在我国HCC发病率还有上升趋势,年龄标化的HCC发病率达31.7/10万人/年.HCC也是恶性程度高、预后很差的一种肿瘤,1995年卫生部统计资料显示HCC死亡率为20.4/10万人,居我国恶性肿瘤致死类型的第二位.