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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 learn the epidemiological characteristics of coronavirus disease 2019 (COVID-19) cases reported from January 21 to February 20,2020 in Zhejiang Province,so as to provide basis for formulating and implementing effective control measures.@*Methods @#The COVID-19 cases reported by Zhejiang Province were extracted from the National Diseases Prevention and Control Information System. A descriptive analysis was adopted for the epidemiological characteristics of COVID-19 cases,including time,spatial and population distribution,severity of symptoms, and exposure history. @*Results @#Totally 1 284 cases were reported,of which 1 176 were confirmed cases (91.59%) and 108 were asymptomatic cases (8.41%). The first confirmed case was reported on January 21. The curve of the disease onset peaked from January 22 to 29, with 87 cases as the biggest number a day. The curve of the disease reported peaked from January 28 to 30, with 130 cases as the biggest number a day. Then the number of the cases showed a declining trend. By February 4, totally 79 counties (cities, districts) had confirmed cases, covering 87.78% of Zhejiang Province, and it has not increase since then. A male to female ratio of 1.02∶1 was reported among 1 284 cases,1 021 were 30-69 years old (79.52%) and 339 (26.40%) were business service providers. There were 11 health workers reported to be infected,but not by occupational exposure. Among the confirmed cases, 1 010 (85.88%) were clinically mild cases. One case died. Before January 23, the cases were mainly imported from Hubei Province, local cases were predominant by the end of January. @*Conclusions @#The COVID-19 epidemic has spread widely in Zhejiang Province and people are generally susceptible. Most cases were clinically mild, and were aged 30-69 years. No health workers infected were due to occupational exposure. The incidence of COVID-19 in Zhejiang Province has turned into a lower level,suggesting that the early prevention and control measures have achieved initial results. With people returning from holiday, precise prevention and control should be put into effect.
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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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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.