Involvement of microRNA-718, a new regulator of EGR3, in regulation of malignant phenotype of HCC cells.

OBJECTIVE: Hepatocellular carcinoma (HCC) is still one of the most common death-related malignancies worldwide. Because the way onset and progression are hidden most, HCC diagnoses are made at an advanced stage, when they are unsuitable for surgical resection. MicroRNAs are a class of small non-coding RNAs, participating in many aspects of cancers. In this study, we tried to establish the role of microRNA-718 (miR-718) in the malignant phenotype of HCC cells and its possible role in HCC diagnosis. METHODS: Here we first used a methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay, Transwell migration and invasion assays, and colony formation assay to evaluate the impact of miR-718 on the malignant phenotypes of HCC cells. Then, we used bioinformatic methods to predict the target gene of miR-718 and used green fluorescence protein (GFP) reporter assay, Western blot, and quantitative real-time polymerase chain reaction (qRT-PCR) to validate the regulation relationship. Finally, we determined the role of the target gene in the HCC phenotype. RESULTS: We found that the expression of miR-718 was significantly reduced in various HCC cell lines and HCC tissues. Re-expression of miR-718 significantly reduced the cellular viability and colony formation ability as well as inhibited the migration and invasion abilities of HCC cell lines. Early growth response protein 3 (EGR3) is a direct target of miR-718 and is negatively regulated by miR-718. EGR3 could increase the viability and proliferation of HCC cells, and promot the migration and invasion of HCC cells. CONCLUSIONS: miR-718 acts as a tumor suppressive microRNA in HCC via regulating the expression of EGR3, which may provide a new diagnostic marker and treatment target for HCC.

Association between circulating inflammatory molecules and alcoholic liver disease in men.

The association between alcoholic liver disease (ALD) and the inflammatory response remains controversial. The aim of this study was to explore this association between ALD and inflammation. We enrolled 214 male participants, who were divided into three age-matched groups: ALD (n = 135), chronic alcohol ingestion without ALD (non-ALD; n = 42), and control (n = 37). The BMI was significantly higher in the ALD group than in the non-ALD and control groups (all P = 0.000). Further, the constituent ratio of the liver inflammatory level was significantly higher in the ALD group than in the non-ALD and control groups (P = 0.002 and P = 0.000, respectively). In addition, the median serum ALT, AST, and GGT levels were significantly higher in the ALD group than in the control group (P = 0.023, P = 0.008, and P = 0.000, respectively); these levels were also significantly higher in the ALD group than in the non-ALD group (P = 0.013, P = 0.010, and P = 0.000, respectively). The median serum CRP level was significantly higher in the ALD group than in the non-ALD and control groups (P = 0.006 and P = 0.000, respectively). Further, the median serum TNF-α level was significantly lower in the ALD group than in the non-ALD and control groups (P = 0.004 and P = 0.000, respectively). The median serum sOX40L and HSP70 levels were significantly lower in the ALD group than in the control group (P = 0.008 and P = 0.018, respectively). In addition, the ALT, AST, and GGT levels were positively correlated with the CRP level (r = 0.211, P = 0.002; r = 0.220, P = 0.001 and r = 0.295, P = 0.000, respectively), and the GGT level was negatively correlated with the TNF-α (r = -0.225, P = 0.001), sOX40L (r = -0.165, P = 0.016), and HSP70 levels (r = -0.178, P = 0.009). Further, the Cr level was negatively correlated with the IL-10 level (r = -0.166, P = 0.015). Logistic regression analysis verified that the BMI (OR  =  1.637, 95%CI: 1.374-1.951, P  =  0.000) and GGT level were significantly higher (OR  =  1.039, 95%CI: 1.020-1.059, P  =  0.000) and that the TNF-α (OR  =  0.998, 95%CI: 0.996-1.000, P  =  0.030) and HSP70 levels were significantly lower (OR  =  1.017, 95%CI: 1.003-1.031, P  =  0.029) in the ALD group than in the non-ALD group. Further, the moderate-to-severe ALD patients had a significantly higher serum CRP level (Or =   1.349, 95%CI: 1.066-1.702, P  =  0.013) and significantly lower HSP60 (OR  =  0.965, 95%CI: 0.938-0.993, P  =  0.014) and HSP70 levels (OR  =  0.978, 95%CI: 0.962-0.995, P  =  0.010) than the mild ALD patients. These results suggest that ALD patients may present with obesity, liver damage, and an imbalanced inflammatory immune response, mainly manifesting as decreased levels of immune inflammatory cytokines. In addition, they suggest that certain liver and kidney function parameters and ALD severity are either positively or negatively correlated with certain inflammatory cytokines. Hence, ALD patients may be at increased risks of obesity- and inflammation-related diseases. Accordingly, to control the inflammatory response, preventative measures for patients with this disease should include weight control and protection of liver and kidney function.

Causes of Thickening of Carotid Intima-Media Thickness in Patients With Alcoholic Liver Disease: A Prospective Observational Study.

Several studies have reported the relationship between alcoholic liver disease (ALD) and carotid intima-media thickness (CIMT). Few studies, however, have investigated the causes of CIMT thickening in patients with ALD. The authors explored the causes of CIMT thickening in patients with ALD. The authors enrolled 152 patients who were stratified into groups: nonthickening CIMT with ALD (group A); thickening CIMT with ALD (group B); nonthickening CIMT without ALD (group C); and thickening CIMT without ALD (group D). The CIMT was significantly different between patients with and without ALD (χ 2= 3.875, P = 0.049). The patients in groups A, B, and C were significantly younger than group D (P = 0.001, 0.036, and 0.001, respectively). The body mass indexes (BMI) in groups A and B were significantly higher than in group C (P = 0.000 and 0.007, respectively). The blood glucose levels in groups B and D were significantly higher than in group C (P = 0.016 and 0.018, respectively). The blood uric acid levels in group B were significantly higher than in groups A, C, and D (P = 0.009, 0.000, and 0.003, respectively). The blood uric acid in group A was significantly higher than in group C (P = 0.002). The serum total cholesterol (TC) levels of patients in group B were significantly higher than in groups A and C (P = 0.027 and 0.000, respectively) and the serum TC level in group A was significantly higher than in group C (P = 0.048). The serum triglyceride (TG) levels in groups A and B were significantly higher than in group C (P = 0.027 and 0.000, respectively). The serum of very low-density lipoprotein (VLDL) levels in group B were significantly higher than in group C (P = 0.000). Although a comparison of the low-density lipoprotein (LDL) and high-density lipoprotein (HDL) serum levels among the 4 groups indicated no changes. The serum LDL levels in group B were significantly higher than in group A (P = 0.008). No significant differences were observed among the groups with respect to serum homocysteine, C-reactive protein (CRP), interleukin-6 (IL-6), malondialdehyde (MDA), superoxide dismutase (SOD), soluble OX40 ligand (sOX40L), or heat shock protein (HSP) 60 or 70. Alcoholic liver disease may result in CIMT thickening. Carotid intima-media thickness is associated with age and metabolic factors in patients with ALD. In addition, ALD might promote the premature occurrence of CIMT thickening. The thickening of carotid artery intima thickness, however, is not associated with cytokine profiles, oxidative balance, or immune responses in patients with ALD.

Changes in tumor necrosis factor-α, heat shock protein 70, malondialdehyde, and superoxide dismutase in patients with different severities of alcoholic fatty liver disease: a prospective observational study.

The relationships among inflammation, oxidative balance, and the severity of alcoholic fatty liver disease (AFLD) remain unknown. The aim of this study is to explore the relationships among tumor necrosis factor alpha (TNF-α), heat shock protein 70 (HSP70), malondialdehyde (MDA), superoxide dismutase (SOD), and the severity of AFLD.From January 2012 to December 2013, 162 participants were enrolled in this study and divided into 4 groups: 44 cases of mild AFLD (group A), 55 cases of moderate-to-severe AFLD (group B), 44 cases of alcohol consumption without AFLD (group C), and 20 cases of no alcohol consumption without AFLD (group D). A cross-sectional study was conducted by detecting the serum levels of TNF-α, HSP70, MDA, and SOD by enzyme-linked immunosorbent assay.The median serum levels of TNF-α and HSP70 among the 4 groups were statistically significant (P = 0.000 and 0.001, respectively). The median serum levels of TNF-α in groups A and B were significantly lower than in group C (P = 0.002 and 0.000, respectively), and the median serum level of TNF-α in group B was significantly lower than in group D (P = 0.023). In addition, the median serum level of HSP70 in group B was significantly lower than in groups A and C (P = 0.002 and 0.000, respectively), and the median serum level of HSP70 in group C was significantly higher than in group D (P = 0.044). However, the median serum level of MDA in group B was significantly lower than only group C (P = 0.008).Chronic alcohol ingestion without AFLD may result in a significant increase in the circulation of certain inflammatory markers; the severity of AFLD is associated with circulating inflammatory markers, and moderate-to-severe AFLD may result in a more significant reduction of these markers. However, moderate-to-severe AFLD may also result in a significant downregulation of oxidative stress products.

Establishing a new animal model for hepadnaviral infection: susceptibility of Chinese Marmota-species to woodchuck hepatitis virus infection.

Hepatitis B virus infection (HBV) is a major medical problem in China. The lack of a suitable infection model in China is recognized as an obstacle for research on HBV in China. Chinese Marmota-species is phylogenetically closely related to Marmota monax, thus, it might be suitable to serve as an animal model for HBV infection. Therefore, we attempted to prove the claim about the existence of woodchuck hepatitis virus (WHV)-like viruses in Chinese Marmota-species and to determine the susceptibility of these species to experimental WHV infection. In the present study, 653 sera from three Chinese Marmota-species, Marmota himalayana, Marmota baibacina and Marmota bobak, were screened for WHV-like viruses by serological and molecular assays. The susceptibility to WHV of three species was investigated by experimental infection and monitored by testing of anti-WHc and WHsAg by ELISA, detection of WHV DNA by PCR, and detection of WHV replication intermediates and antigens in liver samples. No evidence for the existence of a genetically closely related virus to WHV in three Chinese Marmota-species was found by serological assays and PCR. M. himalayana was susceptible to WHV infection as inoculated animals became positive for anti-WHc, WHsAg and WHV DNA. Further, WHV replication intermediates and proteins were detected in liver samples. In contrast, M. baibacina remained negative for tested virological parameters. M. bobak species showed a limited susceptibility to WHV. Our data do not support early reports about WHV-like viruses in China. M. himalayana is suitable for the establishment of a model for hepadnaviral infection.

[Study on experiment of fluorescence spectra detection of organic pesticides in soil].

The fluorescence characteristics of carbamate pesticide, namely carbaryl, was studied based on the basic theory that organic molecules can emit fluorescence as they are excited by rays. Consequently, a fluorescence spectrograph was applied to conduct fluorescence spectrum experiments with standard solution of carbaryl and the hydrolyzed carbaryl, the fluorescence spectra were obtained under the condition of different concentration, and the relation between their fluorescence intensity and concentration was also analyzed. On the basis of this, using the soil as a medium, the fluorescence characteristics of carbaryl and the hydrolyzed carbaryl in the soil were researched as well. The experiment results show that carbaryl and the hydrolyzed carbaryl in the soil can also emit intensive fluorescence which is similar to that in solution as being excited by UV rays. The fluorescence spectra are located between 400 and 750 nm and they all have smooth spectrum forms and fine resolution, so the spectra are suitable for qualitative and quantitative analysis of carbaryl. As a result, it is feasible to carry out the detection and analysis of the concerned pesticides in soil directly or indirectly by fluorescence spectral analysis.

3,9-Dimethyl-2,3-dihydro-phenanthro[1,2-b]furan-4,5-dione.

The title compound, C(18)H(14)O(3), consists of a four-ring system which contains three six-membered rings forming a phenanthrene-dione system and a five-membered 1,2-dihydro-methyl-furan ring. A three-dimensional supra-molecular framework is formed via non-classical inter-molecular C-H⋯O hydrogen bonds.

[Cloning and characterization of genes differentially expressed in human dental pulp cells and gingival fibroblasts].

OBJECTIVE: To study the biological properties of human dental pulp cells (HDPC) by cloning and analysis of genes differentially expressed in HDPC in comparison with human gingival fibroblasts (HGF). METHODS: HDPC and HGF were cultured and identified by immunocytochemistry. HPDC and HGF subtractive cDNA library was established by PCR-based modified subtractive hybridization, genes differentially expressed by HPDC were cloned, sequenced and compared to find homogeneous sequence in GenBank by BLAST. RESULTS: Cloning and sequencing analysis indicate 12 genes differentially expressed were obtained, in which two were unknown genes. Among the 10 known genes, 4 were related to signal transduction, 2 were related to trans-membrane transportation (both cell membrane and nuclear membrane), and 2 were related to RNA splicing mechanisms. CONCLUSION: The biological properties of HPDC are determined by the differential expression of some genes and the growth and differentiation of HPDC are associated to the dynamic protein synthesis and secretion activities of the cell.

[Study on the detection of organic pesticides by fluorescence spectra].

In the present paper the basic theory that organic molecules can emit fluorescence as they are excited by ultraviolet rays is described, the molecular structures of a few common pesticides, such as carbamate, benzoylurea and fungicide, are analyzed, and the mechanism of fluorescence generation is also ascertained. Consequently, the theoretic basis for further detection of pesticides by means of fluorescence methods is provided. Moreover a steady-state fluorescence spectrograph was applied to conduct fluorescence spectrum experiments with standard solutions of these pesticides, the fluorescence spectra were obtained, and their fluorescence characteristics were also analyzed. The results indicate that carbamate, benzoylurea and fungicide pesticides may emit strong fluorescence when excited by UV rays under the condition of a certain solvent, their fluorescence spectra are distinct, and the resolution is fine. As a result, it is feasible to carry out qualitative and the quantitative analysis of the concerned pesticides by fluorescence spectral analysis.

[Study on fluorescence analysis for carbamate pesticides].

In the present paper the basic theory that an organic substance can emit fluorescence as it is excited by ultraviolet rays is described and the relationships between the molecular structures of organic substances and fluorescence generation are studied. According to this theory, the molecular structures of a few common carbamate pesticides such as carbaryl, carbofuran and propoxur etc.are analyzed, their fluorescence characteristics are shown, and the mechanism of fluorescence generation of carbamate pesticides is ascertained. It provides a theoretic basis for further detecting carbamate pesticides by means of a fluorescence analytical method. Moreover a steady-state fluorescence spectrograph is applied to conduct fluorescence spectrum experiments with standard carbaryl solution and carbofuran solution. The results indicate that carbamate pesticides may emit fluorescence as excited by UV under the condition of a certain solvent, and that their fluorescence spectra are distinct and have fine resolution and less interference. It is feasible to detect carbamate pesticides by fluorescence spectral analysis.

[Study on extraction process of zhanjin ruji].

OBJECTIVE: To select the optimum extraction process of Zhanjin Ruji. METHOD: To observe influence of extraction time upon the extraction rate of volatile oil, the orthogonal test was adopted to observe the extraction process by alcohol from the extraction rate and content of the total saponins in Radix Notoginseng. RESULT: The three kinds of herbs including Radix Angelicae Sinensis, Resina Olibani and Myrrha were extracted with water for 3 hours, 95% of volatile oil can be distilled. The three kinds of herbs including Radix Notoginseng, Herba Lycopodii and Radix Gentianae Macrophyllac were extracted by alcohol. Four factors such as alcohol concentration(A), extraction times(B), extraction time(C), and solvent amount(D), had not significant effect on the content of total saponins in Radix Notoginseng in herbal extraction, but factor A and B had significant effect on the extraction rate. The optimum extraction process was as follows extracted with 5 times the amount of the solvent volum 60% alcohol for 3 times and with each time for 1 hour. Three times experiments showed that the extraction rate was 26.5% and the content of the total saponins in Radix Notoginseng was 17.28% mg.g-1. CONCLUSION: The above experimental results can provide experimental basis for deciding the extraction process of Zhanjin Ruji.