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The early diagnosis and effective treatment of hepatocellular carcinoma (HCC) still remains a difficult problem that plagues the medical community. Exosomes are microvesicles with a diameter of 40~100 nm, and contains proteins, lipids and nucleic acids (mRNAs, lncRNAs, circRNAs, and microRNAs). They serve as an information exchange carrier, and play an important role in regulating and controlling the biomolecular function to maintain the stability of the intracellular environment. The function of exosomes in HCC includes intercellular communication, neoangiogenesis, cancer cell metastasis and multidrug resistance, which mediates the transformation of microRNAs (miRNA) and regulate the microenvironment of tumor progression, and then affect the pathophysiological behavior of cancer cells. Exosome-derived miRNA can be used for HCC monitoring or potential specific markers of early diagnosis. In addition, with the development and application prospects it could be a therapeutic goal for HCC. This paper summarizes the recent progress in the study of HCC-derived exosomal miRNA.
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Objective@#To investigate the Wnt3a expression in tissues of HCC and its gene knockout on effects of HepG2 cell proliferation or xenograft tumor growth.@*Methods@#Hepatic Wnt3a expressions in 87 HCC and their matched surrounding tissues were observed by tissue microarray and immunohistochemistry for analyzing its clinicopathological characteristics; Wnt3a-knockout HepG2 cell lines were established by Crispr/cas9-sgRNA system and genomic cleavage efficiency was verified at gene level by surveyor assay. The relative proteins were confirmed by Western blotting; Cell Counting Kit-8 assay was used to examine cell proliferation after knocking-out Wnt3a successfully, and the nude mice HepG2 cell xenograft tumors delete that the relationship between Wnt3a and HCC growth.@*Results@#The positive Wnt3a with brown staining particles was mainly distributed in cytosol and membrane of hepatocytes. The incidence of hepatic Wnt3a expression in cancerous tissues (95.4%) was significantly higher (χ 2 = 47.754, P < 0.001) than that in their surrounding tissues (49.4%). The high Wnt3a expression was 70.1% in the HCC and only 14.9% in the surrounding tissues. High Wnt3a expression was associated with poorly-differentiated grade, liver cirrhosis, HBV infection, portal vein invasion, TNM stage and 5-year survival rate. After knocked-out by Crispr/cas9-sgRNA system successfully, Wnt3a expression was down-regulated significantly at gene or protein level. Key molecule β-catenin in cytoplasma was obviously inhibited. HepG2 cell lines proliferation was suppressed in time-dependent manner. The nude mice HepG2 cell xenograft tumors confirmed that the knock-out of Wnt3a could significantly supressed HCC growth with slower speed (t = 6.418, P < 0.001), smaller volume(869.4 ± 222.5 mm3 vs 355.0 ± 99.9 mm3, t = 5.168, P < 0.001), and lighter weight (0.88 ± 0.20 g vs 0.35 ± 0.11 g, t = 5.628, P < 0.001)compared with the control group.@*Conclusion@#Abnormal expression of Wnt3a could be expected as a promising target for HCC gene therapy.
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Objective@#To quantitatively detect CD44 expression in patients with nonalcoholic fatty liver disease (NAFLD) for comparative analysis.@*Methods@#Patients with chronic liver diseases accompanied with or without NAFLD, including chronic hepatitis B, cirrhosis and hepatocellular carcinoma after chronic hepatitis B, and healthy blood donors as normal controls who admitted to the Affiliated Hospital of Nantong University from May to October 2018 were selected. The proportion of CD44 positive cells was analyzed by flow cytometry. CD44 level was quantified by an enzyme-linked immunosorbent assay, and the biochemical indicators such as serum aspartate aminotransferase, alanine aminotransferase activity, total cholesterol and triglyceride were routinely analyzed. The cancerous and adjacent cancerous tissues of patients accompanied with or without NAFLD were collected by self-matching method and analyzed by immunoblotting and histochemistry and compared by CD44 integrated optical density. Image-Pro Plus version 6.0, Image J, GraphPad Prism 5.0, Photoshop, Microsoft Excel and IBM SPSS statistics 23 were used to analyze and draw pictures. An independent sample t-test was used to compare the differences between groups.@*Results@#Patients accompanied with NAFLD had hepatocyte injury and dyslipidemia. NAFLD and chronic liver disease patients had significantly elevated serum CD44 levels than normal control group (P < 0.01). CD44 positive lymphocyte ratio was 78.19 % ± 16.33 % in NAFLD patients and 68.47% ± 20.91% in chronic hepatitis B group, which was higher than the control group (46.51% ± 20.52%). Chronic hepatitis B group with steatosis had significantly higher CD44 concentration (181.42 ± 49.36) ng/ml than chronic hepatitis B group (142.52 ± 53.87) ng/ml and normal control group (99.47 ± 15.23) ng/ml. CD44/GAPDH ratio in the liver cancer group (1.306 ± 0.614) was significantly higher than paracancerous group (0.477 ± 0.291) and the difference between the two groups was statistically significant (t = 3.451, P = 0.004). The integrated optical density of CD44 in the NAFLD-related liver cancer and paracancerous group were 25.721 ± 5.881 and 14.155 ± 4.001 and the difference between the groups was statistically significant (t = 14.544, P < 0.001). The pathological features of high expression of CD44 in patients with hepatocellular carcinoma were significantly correlated with HBV infection, tumor size, single/multi-center, and lymph node metastasis, degree of differentiation, TNM grade, Child-Pugh score, portal vein tumor thrombus and extrahepatic metastasis. HCC patients with NAFLD had significantly higher serum CD44 (234.62 ± 69.40) ng/ml than patients without NAFLD (186.49 ± 58.89) ng/ml (t = -3.191, P = 0.002), but there was no statistically significant difference in the clinicopathological characteristics between the high/low CD44 groups of HCC patients with NAFLD.@*Conclusion@#The results suggest that CD44 is abnormally activated and its mechanism may play an important role in the progression of NAFLD.
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The biological functions of high-mobility group (HMG) proteins include regulation of DNA replication, transcription, recombination and repair. According to molecular weight, sequence alignment and DNA structural characteristics, HMG proteins are subdivided into three superfamilies (HMGA, HMGB and HMGN). Recently, HMGB family members (HMGB1, HMGB2, HMGB3, and HMGB4) found to interact with hepatitis B or C virus. Therefore, activation of relevant signaling molecules to regulate transcription of genes related to hepatocellular carcinoma as a mediator of inflammation promoting HCC progression has attracted considerable attentions. This article focuses on the clinical application of the expression of HMGB family members in the process of HCC progression.
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Hepatocellular carcinoma is one of the most common digestive system tumors. Its occurrence and development are considered as multi-factorial and multi-step process. Recent studies have shown that wingless-related integration (Wnt) pathway plays an important role in the HCC progression and is associated with malignant transformation of hepatocytes, HCC metastasis, drug resistance and liver cancer stem cells. This article analyzes the expression of key signaling molecules in Wnt pathway and its value in diagnosis, prognosis and targeted therapy, and outlines the research progress of Wnt pathway targeted drugs for the treatment of HCC, with a view to providing targeted therapy research for HCC reference.
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The monitoring of malignant transformation of hepatocytes or early diagnosis of primary hepatocellular carcinoma (PHC) remains a challenge in the medical world. Routine examinations including serum alpha-fetoprotein level and ultrasound examination have a limited value in the diagnosis of small hepatocellular carcinoma; however, the effective treatment of PHC depends on its early diagnosis. In recent years, molecular markers including important signaling molecules in PHC-related pathways, carcinoembryonic proteins, and non-coding RNA help with the monitoring of malignant transformation of hepatocytes or early diagnosis of liver cancer. This article reviews the valuable molecular markers in the monitoring of malignant transformation of hepatocytes or early diagnosis of liver cancer.
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Objective@#To investigate the dynamic expression of hepatic carnitine palmitoyltransferase-II (CPT-II) in the mitochondrial inner membrane during hepatocyte malignant transformation induced by lipid accumulation.@*Methods@#Male Sprague-Dawley rats were divided randomly into control, fatty liver, and induced cancer groups, which were fed with normal, high-fat (HF), and HF containing 2-fluorenylacetamide (0.05%, 2-FAA) diets, respectively, for 14 weeks. One rat from each group was sacrificed every two weeks and the blood and liver samples were collected. Liver morphological changes were evaluated with hematoxylin and eosin staining, and the liver tissue samples were divided into control, fatty liver, degeneration, precancerous, and cancerous groups accordingly. Hepatic lipids were dyed by the oil red O method. The CPT-II expression was measured by immunohistochemistry and compared with the specific CPT-II concentration (ng/mg liver protein, ng/mg P) among different groups. Serum levels of circulating total cholesterol (Tch), triglyceride (TG), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) were quantitatively analyzed.@*Results@#Massive lipid accumulation hepatocytes was seen in rats on HF and HF containing 2-FAA diets. The lipid levels in the control group were significantly lower than those in the fatty liver (t = -11.556, P < 0.001), degeneration (t = -4.847, P = 0.04), precancerous (t = -13.652, P = 0.005), and cancerous groups (t = -10.896, P = 0.008). The serum TG and Tch levels in the degeneration, precancerous, and cancerous groups were 2-3 times higher than those in the control group (P < 0.05). After 2-FAA treatment, the morphological changes of rat hepatocytes showed the progression from degeneration and precancerosis to cancerosis, with hepatocyte injury. The serum AST and ALT levels in the degeneration, precancerous, and cancerous groups were significantly higher (4-8 times) than those in the control group (P < 0.05). The specific concentration of liver CPT-II expression was significantly reduced during hepatocyte malignant transformation, as confirmed by immunohistochemistry, with the CPT-II levels significantly lower in the cancerous group than in any of other groups (P < 0.05).@*Conclusion@#Low hepatic CPT-II expression might lead to abnormal lipid accumulation in hepatocytes, which should promote the malignant transformation of hepatocytes.
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Hepatocellular carcinoma (HCC) is still one of common malignant cancers worldwide, with increasing incidence and mortality rates. Early diagnosis and effective treatment for HCC remain to be explored. This article introduces the research advances in the early specific diagnosis and effective therapies for HCC in 2016, such as molecular markers in the specific diagnosis and targeted therapy for HCC, main therapeutic regimens, robot-assisted liver resection, and no-touch radiofrequency ablation.
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Nonalcoholic fatty liver disease (NAFLD) is a clinicopathological syndrome characterized by diffuse macrovesicular steatosis and excessive lipid accumulation in the liver due to nonalcoholic reasons and other specific factors for liver injury, including simple fatty liver, nonalcoholic steatohepatitis, and liver cirrhosis. With the increasing incidence of NAFLD, the risk of hepatocellular carcinoma (HCC) is also increasing, but the exact mechanism remains unknown. This paper summarizes the roles of lipid accumulation, related cells, proteins, hormones, and metabolites in malignant transformation of hepatocytes, as well as the relationship between NAFLD, HCC, and lipid metabolism, and provides a new perspective for the prevention of HCC in clinical practice.
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<p><b>OBJECTIVE</b>To investigate the inhibitory effects of intervention of the tumor necrosis factor-alpha (TNFa)/nuclear factor-kappa B (NF-kappaB) signaling pathway activation on hepatoma cell proliferation and to explore its mechanism.</p><p><b>METHODS</b>A rodent hepatoma model was established by feeding N-2-fluorenylacetamide (2-N-FAA) to male Sprague-Dawley rats. Human subjects with various liver diseases were enrolled in the study, and serum and peripheral blood nuclear cells were collected for analysis. HepG2 cells were cultured in vitro and treated with anti-TNFa (monoclonal antibody, mAb) to down-regulate its expression or transfected with siRNA targeting the p65 subunit of NF-kappaB to inhibit its activation. The liver cell line L02 was used as a control. Changes in protein and gene expression levels of NF-kappaB and TNFa were analyzed by Western blotting or enzyme-linked immunosorbent assay and real-time PCR, respectively. Changes in the cell cycle or apoptosis were evaluated by flow cytometry or Annexin-V/PI double-labeling assay, respectively.</p><p><b>RESULTS</b>TNFa and NF-kappaB expression showed increasing trends during the malignant transformation of rat hepatocytes, and the differential expression patterns showed association with histopathological alterations in the hepatocytes. Following treatment with the TNFa mAb, the HepG2 cells showed a higher percentage of apoptotic cells than the untreated control cells (21.45% +/- 4.07% vs. 5.63% +/- 0.93%, q =10.07, P less than 0.01).There was a significant difference in the rate of cells in the G0/G1 phase in the p65-siRNA transfected cells (66.23% +/- 1.29% vs. untreated control cells: 59.00% +/- 1.02%, q =10.98, P less than 0.01). The decreased expression of TNFa and NF-kappaB in cell culture supernatants was positively correlated with the dose of treatment (r =0.89, P less than 0.01), with the most robust decreases being achieved with the highest concentrations ( P less than 0.01). NF-kappaB expression was significantly higher in the HepG2 cells than in the L02 cells, and transfection of p65-siRNA reduced the mRNA (93%) and protein (62%) levels and increased the cell apoptosis index (to 85%).</p><p><b>CONCLUSION</b>Proliferation of hepatoma cells may be significantly inhibited by intervening in the activation of the TNFa/NF-kappaB signaling pathway, which promotes cell apoptosis and blocks cell cycling.</p>
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Adult , Aged , Animals , Female , Humans , Male , Middle Aged , Rats , Apoptosis , Carcinoma, Hepatocellular , Pathology , Cell Proliferation , Hep G2 Cells , Hepatocytes , Metabolism , Liver Neoplasms , Pathology , NF-kappa B , Metabolism , RNA, Messenger , Genetics , Rats, Sprague-Dawley , Signal Transduction , Transfection , Tumor Necrosis Factor-alpha , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To investigate the effects of Annexin A2 (ANXA2) deficiency on the malignant biological behaviour of hepatoma cells.</p><p><b>METHODS</b>The human hepatocellular carcinoma (HCC) cells lines MHCC97-H, HepG2, SMMC-7721, SMMC-7402 and L02 were evaluated. The expression and distribution of ANXA2 were analysed by western blotting, real-time PCR, immunofluorescence and immunohistochemistry.Cell cycle was assessed by flow cytometry and propidium iodide staining. Effects of ANXA2 silencing on invasion and migration potential were assessed by transwell assay and wound healing assay, respectively. Proliferative potential was assessed by CCK-8 kit in vitro and xenograft tumour-growth assay in vivo. The t-test, chi square test, rank sum test, q-test and F-test were used for statistical analyses.</p><p><b>RESULTS</b>The expression level of ANXA2 was markedly higher in the MHCC97-H cells with high metastasis potential than in the HepG2, SMMC-7721, SMMC-7402 and L02 cells. The efficiency of shRNA-mediated ANXA2 deficiency was more than 80%. Immunofluorescence analysis of the MHCC97-H cells indicated that ANXA2 expression was mainly localized to the cellular membrane and cytoplasm, with some nuclear localization. Down-regulation of ANXA2 led to S-phase arrest of HCC cells (q =8.001, P =0.002) and an inhibition of proliferation (q =17.140, P less than 0.01), migration (q =12.808, P less than 0.01) and invasion potential (q =9.069, P =0.002). Xenograft tumour-growth assay indicated that shRNA targeting of ANXA2 led to lower tumour weight (q =11.968, P < 0.001) and down-regulated ANXA2 expression (Z =2.530, P =0.011).</p><p><b>CONCLUSION</b>Down-regulation of Annexin A2 gene transcription effectively changes the biological behaviours of hepatoma cells, and may represent a potential target of HCC molecular therapies.</p>
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Animals , Humans , Annexin A2 , Genetics , Carcinoma, Hepatocellular , Pathology , Cell Line, Tumor , Cell Proliferation , Liver Neoplasms , Pathology , Neoplasm Transplantation , RNA, Small Interfering , Genetics , Signal Transduction , Transcription, GeneticABSTRACT
During years of managent practice of postgraduate training,it is found that great attention of leadership is a good foundation,that pre-service training is an essential part,that srtict implementation of the requirements on postgraduate,advisor-responsibility system unified instruction under subject,focusing on process-management of training and exploring actively effective incentive training methods is necessary.
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With the electronic medical records information system as the core of hospital information platform design, this paper introduces the design of the system which is structured for electronic medical records, and the advantage and effectiveness is also introduced. From the aspects of the concept, system framework, data integration, right frame and safety control, identity index and a clinical pathway, etc, the paper introduces the basic idea and process of the hospital information platform design, with the hospital recording electronic medical records as the core.
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Electronic Health Records , Hospital Information Systems , Medical Records Systems, ComputerizedABSTRACT
Objective To explore the clinical value of peripheral blood albumin gene and alpha-fetoprotein gene in early diagnosis and differential diagnosis ofhepatocellular carcinoma(HCC).Methods Two gene fragments were amplified by nested reverse transcription polymerase chain reaction(RT-PCR)with the samples from patients.Results The positive tales of peripheral blood gene fragments from patients with HCC were 53.7％in AFP and 59.7％in Alb.respectively.The former Wag signifieantly higher than that in liver cirrhosis(LC),chronic hepatitis(CH),acute hepatitis(AH)and extrahepatic tumor(ET).The latter Wag also significantly higher than that in LC,CH and ET(P<O.01),but there were no remarkably difference in the positive rate of Alb.mRNA between HCC and AH.The presenee of AFP-mRNA and AIb-mRNA were correlmed with the stage of HCC and intrahepatic metastasis.Conclusion The results suggest that gene fragments of peripheral blood AFP and Alb are useful marker8 for diagnosis of HCC,differentiating extrahepatic metastasis of HCC and monitoring recurrence after HCC operation.
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BACKGROUND: Cardiospecific proteins of the troponin-tropomyosin complex in the contractile system of the cardiomyocytes have challenged creatine kinase isoenzyme MB (CK-MB) as the "gold standard" for the early biochemical detection of acute myocardial injury.OBJECTIVE: To investigate cardiac troponin Ⅰ messager RNA (cTnI-mRNA) in peripheral blood and its clinical values in diagnosis of patients with myocardial injury.DESIGN: A basic and observational study for set up a method to analyze cTnI-mRNA.SETTING: Department of Cardiology, Affiliated Hospital, Nantong University.MATERIALS: The project was accomplished from May 2003 to May 2005 in Research Center of Clinical Molecular Biology, and Department of Pathology, Affiliated Hospital, Nantong University. The cTnI-mRNA was detected from blood by a nested PCR assay, and its clinical values as a sensitive myocardial diagnostic marker were confirmed in patients with myocardial injury.METHODS: Pathologic features and microstructure of cardiac myocytes were examined by H&E staining or electron microscopy. The cTnI-mRNA was extracted from blood and synthesized to cDNA through random primers and reverse transcriptase, and amplified by a nested PCR assay, and its clinical values as a myocardial diagnostic marker were investigated in patients with myocardial injury.MAIN OUTCOME MEASURES: Microstructure of cardiomyocytes, sensitivity of analysis method and diagnostic values.RESULTS: Microstructure of cardiomyocytes with mitochondria swell,rupture, vacancy-like denaturation, nucleus abnormality, and chromatin condensed were observed by electron microscopy. The cTnI-mRNA fragments from heart and blood were successfully amplified and the sensitivity was 2 pg/μL. The product sequences from tissues or blood were confirmed by sequencing. The cTnI-mRNA from cardiac myocytes was found that it present in blood plasma and not in circulating nucleus cell. The incidence of blood cTnI-mRNA of chronic cardiomyopathy was significantly higher (P ＜ 0.05) than that of serum enzymatic patterns or cTnI quality,respectively.CONCLUSION: The analysis of blood cTnI-mRNA is a sensitive marker for diagnosis and monitoring of myocardial injury.
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Objective To explore the clinical significances of total antioxidation (TAO) and nitric oxide synthase (NOS) expression in patients with liver diseases.Methods The concentrations of TAO,NOS and nitric oxide(NO) were determined and the mechanism of their changes were analyzed in sera of patients with acute hepatitis(AH),chronic hepatitis(CH),liver cirrhosis(LC) and hepatocellular carcinoma(HCC).Results The abnormal rate of TAO was 80% in AH or in CH,and 50% in LC or in HCC,respectively.The sera NOS activity over normal reference value was 70% in patients with liver diseases.The abnormal rate of NO level was 70% in AH,CH and LC groups,and 48% in HCC,respectively.The average level of serum TAO was significantly higher in AH group or in CH group,but not in both HCC and LC groups than that in normal subjects.The average levels of NO and NOS were significantly higher in patients with liver diseases than those in normal subjects.However,the two markers were lower in HCC patients than those in AH,CH and LC groups,respectively.Conclusions The data suggest that the activity of NOS is close relation to serum NO level in patients with liver diseases,the increase of NO concentration may play a role in protection of hepatocytes.
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Objective To explore the roles of vascular endothelial growth factor (VEGF) in microvessel angiogennesis, development and metastasis of hepatocellular carcinoma (HCC). Methods The expression and cellular distributions of VEGF in HCCs were investigated by immunohistochemical method, and the levels of total RNA and VEGF also were quantitatively analysed in HCCs and paracancerous tissues. Results The positive rates of VEGF were 63 9% in all HCCs, 78 3% in non-encapsuled HCCs, and 90 9% in HCCs with extra-hepatic metastasis, respectively. VEGF expression was not associated with tumor size and differentiation degree. The total RNA level in HCCs was significantly lower than that in paracancerous and distal paracancerous tissues (P
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There have emerged gradually serious one-sided benefits chasing behaviour over the past 10 years in many hospitals. In this article we've tried to analyze its scocial roots and disclose its harming effects which lead us to ethical misunderstanding and contradict serving tenets in our clinical practice. We also present these pertinent criteria of medical ethics and search after approaches for keeping away these mistakes repeatedly.