VCP/p97, Down-Regulated by microRNA-129-5p, Could Regulate the Progression of Hepatocellular Carcinoma

Valosin containing protein (VCP)/p97 plays various important roles in cells. Moreover, elevated expression of VCP in hepatocellular carcinoma (HCC) is correlated with increased incidence of recurrence. But the role of VCP in HCC progression in vitro and in vivo is unclear. And there are few reports about the regulation mechanism on the expression of VCP in HCC. In this study, it was identified that the level of VCP was frequently increased in human HCC tissues. In addition, down-regulation of VCP with siRNAs could dramatically suppress the genesis and progression of tumor in vivo. It was found that miR-129-5p directly inhibited the expression of VCP in several HCC cell lines. Meanwhile, the level of VCP in HCC tissues was negatively associated with the level of miR-129-5p. Our further investigation showed that the enhanced expression of miR-129-5p also suppressed tumor growth in vivo. Moreover, it was revealed that miR-129-5p could inhibit the degradation of IκBα and increase the apoptosis and reduce the migration of HCC cells by suppressing the expression of VCP. Our results revealed that the expression of VCP was directly regulated by miR-129-5p and this regulation played an important role in the progression of HCC

Purification Effect of Oysters Based on the Analysis of Environmental Parameters

With the development of coastal industries, marine pollution has intensified, and more and more shellfish in marine areas have been polluted to varying degrees. People's health can be compromised when eating contaminated shellfish. In the actual purification process of the shellfish purification plant project, there is no in-depth exploration of the parameters of the purification pond. Based on supplementing the basic research for the development of the oyster purification industry at different temperatures, salinities, pH, ultraviolet lamp power, irradiation time, and water flow rate, we find the best environmental conditions are a temperature of 30 °C, a pH of 8, a salinity of 32%, an ultraviolet lamp illumination power of 9w, an ultraviolet lamp irradiation time of 12h, and a water body flow rate of 0.5m/s. We supplement basic research related to bioengineering in China. We hope that our results can provide reference for further research and contribute to the sustainable development of the oyster purification industry. Create economic benefits for society

Discrimination and Characterization of the Volatile Organic Compounds in <i>Schizonepetae Spica</i> from Six Regions of China Using HS-GC-IMS and HS-SPME-GC-MS

Volatile organic compounds (VOCs) are the main chemical components of Schizonepetae Spica (SS), which have positive effects on the quality evaluation of SS. In this study, HS-SPME-GC-MS (headspace solid-phase microextraction-gas chromatography-mass spectrometry) and HS-GC-IMS (headspace-gas chromatography-ion mobility spectrometry) were performed to characterize the VOCs of SS from six different regions. A total of 82 VOCs were identified. In addition, this work compared the suitability of two instruments to distinguish SS from different habitats. The regional classification using orthogonal partial least squares discriminant analysis (OPLS-DA) shows that the HS-GC-IMS method can classify samples better than the HS-SPME-GC-MS. This study provided a reference method for identification of the SS from different origins

Response of Organic Fertilizer Application to Soil Microorganisms and Forage Biomass in Grass–Legume Mixtures

It has been widely recognized that organic fertilizer (OF) application under monoculture and continuous cropping can change the microbial community and increase forage biomass in the Qinghai–Tibet Plateau. However, as a commonly used grassland planting pattern, the way in which grass–legume mixtures respond to OF application remains unclear. To clarify application effects of organic fertilizer in the grass–legume mixtures, we conducted a field experiment at the Qinghai–Tibet Plateau and collected the rhizospheric and bulk soils to reveal their microbial community by using high-throughput sequencing and molecular ecological networks. It was found that OF application changed the microbial community and increased the forage biomass under monoculture. However, in grass–legume mixtures, we found that OF application did not promote the increase of forage (Gramineae) biomass (Student t-test: p > 0.05). By analyzing both prokaryote and fungal communities, it was found that OF application had a greater impact on bulk soil microorganisms than on those of the rhizosphere in grass–legume mixtures. Co-occurrence network analysis showed that the rhizosphere and bulk soil networks of grass–legume mixtures were significantly more vulnerable under OF treatment (vulnerability of prokaryotes in grass: 0.1222; vulnerability of prokaryotes in legumes: 0.1730; fungal vulnerability in grass: 0.0116; fungal vulnerability in legumes: 0.0223) than non-OF treatment (vulnerability of prokaryotes in grass: 0.1015; vulnerability of prokaryotes in legumes: 0.1337; fungal vulnerability in grass: 0.0046; fungal vulnerability in legumes: 0.0126), which indicated that OF application did not provide favorable conditions for microbial interactions in grass–legume mixtures. In addition, structural equation modeling showed that OF application had some significant negative impacts on soil physicochemical properties and the robustness of the prokaryote community. The robustness of fungi had a significant negative (p p > 0.05) direct impact on the forage biomass, which indicated that the OF did not promote forage biomass in grass–legume mixtures. These results suggest that the application of organic fertilizer is unnecessary for grass–legume mixtures, because it does not promote the interactions between rhizospheric microbes and forage

Response of Organic Fertilizer Application to Soil Microorganisms and Forage Biomass in Grass&ndash;Legume Mixtures

It has been widely recognized that organic fertilizer (OF) application under monoculture and continuous cropping can change the microbial community and increase forage biomass in the Qinghai&ndash;Tibet Plateau. However, as a commonly used grassland planting pattern, the way in which grass&ndash;legume mixtures respond to OF application remains unclear. To clarify application effects of organic fertilizer in the grass&ndash;legume mixtures, we conducted a field experiment at the Qinghai&ndash;Tibet Plateau and collected the rhizospheric and bulk soils to reveal their microbial community by using high-throughput sequencing and molecular ecological networks. It was found that OF application changed the microbial community and increased the forage biomass under monoculture. However, in grass&ndash;legume mixtures, we found that OF application did not promote the increase of forage (Gramineae) biomass (Student t-test: p &gt; 0.05). By analyzing both prokaryote and fungal communities, it was found that OF application had a greater impact on bulk soil microorganisms than on those of the rhizosphere in grass&ndash;legume mixtures. Co-occurrence network analysis showed that the rhizosphere and bulk soil networks of grass&ndash;legume mixtures were significantly more vulnerable under OF treatment (vulnerability of prokaryotes in grass: 0.1222; vulnerability of prokaryotes in legumes: 0.1730; fungal vulnerability in grass: 0.0116; fungal vulnerability in legumes: 0.0223) than non-OF treatment (vulnerability of prokaryotes in grass: 0.1015; vulnerability of prokaryotes in legumes: 0.1337; fungal vulnerability in grass: 0.0046; fungal vulnerability in legumes: 0.0126), which indicated that OF application did not provide favorable conditions for microbial interactions in grass&ndash;legume mixtures. In addition, structural equation modeling showed that OF application had some significant negative impacts on soil physicochemical properties and the robustness of the prokaryote community. The robustness of fungi had a significant negative (p &lt; 0.001) impact on forage biomass, but OF application had no significant (p &gt; 0.05) direct impact on the forage biomass, which indicated that the OF did not promote forage biomass in grass&ndash;legume mixtures. These results suggest that the application of organic fertilizer is unnecessary for grass&ndash;legume mixtures, because it does not promote the interactions between rhizospheric microbes and forage

miR-129-5p could directly regulate the expression of VCP.

<p>A: miR-129-5p could significantly suppress the luciferase activities of pGL3-VCP-3′UTR in HepG2 cells. Five predicted miRNAs (miR-103, miR-107, miR-129-5p, miR-136, miR-339-5p) were transfected in HepG2 cells, respectively, with VCP 3′UTR report. 72 h after the transfection, the cells were harvested for luciferase activity. Shown data are representative from three independent experiments. **:P<0.01. B: Bioinformatic analysis of miR-129-5p predicted binding sites in the VCP 3′UTR. There were two putative miR-129-5p target sites located in the VCP 3′UTR (162–168 and 505–511). Three mutant reporter vectors were constructed with the deletion of two target sites individually (pGL3-VCP-3′UTRm1, pGL3-VCP-3′UTRm2) or both (pGL3-VCP-3′UTRm3). C: The luciferase activity of the mutant VCP 3′UTR report genes were not regulated by miR-129-5p in HepG2 cells. Shown data are representative from three independent experiments.**:P<0.01. D: miR-129-5p could significantly suppress the luciferase activities of pGL3-VCP-3′UTR in MHCC-LM3 cells, while has no effect on three mutant reporter vectors. Shown data are representative from three independent experiments.**:P<0.01. E: The inhibiter of miR-129-5p could increase the luciferase activities of pGL3-VCP-3′UTR in SK-HEP1 cells, while has no effect on three mutant reporter vectors. Shown data are representative from three independent experiments. **:P<0.01. F: Analysis of the level of miR-129-5p in HCC tissues by qRT-PCR. The result showed that the level of miR-129-5p is frequently reduced in human HCC tissues.*:P<0.05.**:P<0.01. G: The level of miR-129-5p was negatively correlated with the expression of VCP in HCC. The 39 tissue samples of HCC were divided into two groups according to the level of VCP. The level of miR-129-5p in each sample from the two groups (level 1,n = 17;level 2,n = 22) was measured by qRT-PCR and compared. U6 snRNA was used as internal control gene. Dot plots showed an inverse relationship between U6 snRNA and miR-129-5p expression in HCC. Significant differences were determined using Student's t tests. **: P<0.01; Bars: mean±SD.</p