CRISPR/Cas9-mediated knockout of the Vanin-1 gene in the Leghorn Male Hepatoma cell line and its effects on lipid metabolism

Xu Lu,Wang Zhongliang,Liu Shihao,Wei Zhiheng,Yu Jianfeng,Li Jun,Li Jie,Yao Wen,Gu Zhiliang 아세아·태평양축산학회 2024 Animal Bioscience Vol.37 No.3

Objective: Vanin-1 (VNN1) is a pantetheinase that catalyses the hydrolysis of pantetheine to produce pantothenic acid and cysteamine. Our previous studies have shown that the VNN1 is specifically expressed in chicken liver which negatively regulated by microRNA-122. However, the functions of the VNN1 in lipid metabolism in chicken liver haven’t been elucidated. Methods: First, we detected the VNN1 mRNA expression in 4-week chickens which were fasted 24 hours. Next, knocked out VNN1 via CRISPR/Cas9 system in the chicken Leghorn Male Hepatoma cell line. Detected the lipid deposition via oil red staining and analysis the content of triglycerides (TG), low-density lipoprotein-C (LDL-C), and highdensity lipoprotein-C (HDL-C) after VNN1 knockout in Leghorn Male Hepatoma cell line. Then we captured various differentially expressed genes (DEGs) between VNN1- modified LMH cells and original LMH cells by RNA-seq. Results: Firstly, fasting-induced expression of VNN1. Meanwhile, we successfully used the CRISPR/Cas9 system to achieve targeted mutations of the VNN1 in the chicken LMH cell line. Moreover, the expression level of VNN1 mRNA in LMH-KO-VNN1 cells decreased compared with that in the wild-type LMH cells (p<0.0001). Compared with control, lipid deposition was decreased after knockout VNN1 via oil red staining, meanwhile, the contents of TG and LDL-C were significantly reduced, and the content of HDL-C was increased in LMH-KO-VNN1 cells. Transcriptome sequencing showed that there were 1,335 DEGs between LMH-KO-VNN1 cells and original LMH cells. Of these DEGs, 431 were upregulated, and 904 were downregulated. Gene ontology analyses of all DEGs showed that the lipid metabolism-related pathways, such as fatty acid biosynthesis and long-chain fatty acid biosynthesis, were enriched. KEGG pathway analyses showed that “lipid metabolism pathway”, “energy metabolism”, and “carbohydrate metabolism” were enriched. A total of 76 DEGs were involved in these pathways, of which 29 genes were upregulated (such as cytochrome P450 family 7 subfamily A member 1, ELOVL fatty acid elongase 2, and apolipoprotein A4) and 47 genes were downregulated (such as phosphoenolpyruvate carboxykinase 1) by VNN1 knockout in the LMH cells. Conclusion: These results suggest that VNN1 plays an important role in coordinating lipid metabolism in the chicken liver.

Effect of MgO on migration and distribution of chromium in stainless-steel slag glass–ceramics and glass properties

Zhifang Tong,Congcong Xu,Jiaxing Wang,Zhiheng Jia 한양대학교 청정에너지연구소 2023 Journal of Ceramic Processing Research Vol.24 No.1

Herein, stainless-steel slag was used to prepare glass–ceramics. The effects of the MgO content on the occurrence, distributionstate, and migration behavior of Cr in glass–ceramics as well as glass properties were systematically studied. Results revealedthat in the nucleation stage, the number of Cr-spinel (Cr spinel) nanocrystals initially increased and then decreased with theincreasing MgO content. In the crystallization stage, the diopside crystal phase considered Cr-spinel nanocrystals asheterogeneous nuclei, which gradually nucleated and grew. Transmission electron microscopy and X-ray photoelectronspectroscopy analyses showed that part of Cr in the spinel migrated into the diopside lattice and some other parts remainedin the Cr spinel wrapped by the diopside. Cr in the glass phase also diffused and migrated into the diopside lattice with theformation of diopside crystals. When the MgO content was 12.3 wt.%, 97.33 wt.% of the total amount of Cr was observedin the diopside crystal phase and the leaching concentration of Cr in the glass–ceramics was only 0.006 mg/L. The compressivestrength of the glass–ceramics was 261.7 MPa, and the Vickers hardness was 1007.7 HV. The research results providetheoretical and technical support for strengthening Cr fixation and realizing harmless and high-value utilization of stainlesssteelslag.

Lipidomic profiling of Skipjack tuna (Katsuwonus pelamis) by ultrahigh-performance liquid chromatography coupled to high resolution mass spectrometry

Hu, Lingping,Hu, Zhiheng,Chin, Yaoxian,Yu, Haixia,Xu, Jianhong,Zhou, Jianwei,Liu, Donghong,Kang, Mengli,Hu, Yaqin The Korean Society of Fisheries and Aquatic Scienc 2022 Fisheries and Aquatic Sciences Vol.25 No.3

A method of ultrahigh performance liquid chromatography coupled to high resolution mass spectrometry (UPLC-HRMS) was established for characterization of the lipid profile of Skipjack tuna. Over 300 lipid molecular species were identified through cross-acquisition in both positive and negative ion mode. Phospholipids (PLs) were dominant in Skipjack tuna. Lysophosphatidylethanolamine (LPE), phosphatidylethanolamine (PE), lysophosphatidylcholine (LPC) and phosphatidylcholine (PC) were the main lipid molecular species in PLs, accounting for 89.24% of the total PLs. The ratio of sphingolipids (SLs) and glycerolipids (GLs) were considerable, accounting for 12.30% and 13.60% of the total lipids respectively. Ceramide (Cer) was the main lipid molecular species of SLs, accounting for 64.96% of total SLs, followed by sphingomyelin (SM), accounting for 25.45% of total SLs. Ether diglycerides (ether DG) were the main lipid molecular species of GLs (97.83%). The main fatty acids (FAs) are unsaturated fatty acids (UFAs) in Skipjack tuna. Besides, a new FAs class branched fatty acid esters of hydroxy fatty acids (FAHFA) was detected, together with the FA. The active lipids identified in this study can be used to evaluate the nutritional value of Skipjack tuna.

Neutron-irradiated effect on the thermoelectric properties of Bi2Te3-based thermoelectric leg

Zhao Huanyu,Liu Kai,Xu Zhiheng,Liu Yunpeng,Tang Xiaobin 한국원자력학회 2023 Nuclear Engineering and Technology Vol.55 No.8

Thermoelectric (TE) materials working in radioisotope thermoelectric generators are irradiated by neutrons throughout its service; thus, investigating the neutron irradiation stability of TE devices is necessary. Herein, the influence of neutron irradiation with fluences of 4.56 1010 and 1 1013 n/cm2 by pulsed neutron reactor on the electrical and thermal transport properties of n-type Bi2Te2.7Se0.3 and ptype Bi0.5Sb1.5Te3 thermoelectric alloys prepared by cold-pressing and molding is investigated. After neutron irradiation, the properties of thermoelectric materials fluctuate, which is related to the material type and irradiation fluence. Different from p-type thermoelectric materials, neutron irradiation has a positive effect on n-type Bi2Te2.7Se0.3 materials. This result might be due to the increase of carrier mobility and the optimization of electrical conductivity. Afterward, the effects of p-type and n-type TE devices with different treatments on the output performance of TE devices are further discussed. The positive and negative effects caused by irradiation can cancel each other to a certain extent. For TE devices paired with p-type Bi0.5Sb1.5Te3 and n-type Bi2Te2.7Se0.3 thermoelectric legs, the generated power and conversion efficiency are stable after neutron irradiation.

Xylazole inhibits NO-cGMP pathway in fetal rat nerve cells

Xinyu Wang,Yue Wu,Lin Liu,Hui Bai,Zhiheng Zhang,Mingchao Zhao,Tianwen Ma,Xiaopeng Song,Lina Jia,Liangyu Lv,Yue Yu,Xinyu Xu,Hong Chen,Li Gao 대한수의학회 2022 Journal of Veterinary Science Vol.23 No.1

Background: Xylazole (Xyl) is a veterinary anesthetic that is structurally and functionally similar to xylazine. However, the effects of Xyl in vitro remain unknown. Objectives: This study aimed to investigate the anesthetic mechanism of Xyl using fetal rat nerve cells treated with Xyl. Methods: Fetal rat nerve cells cultured for seven days were treated with 10, 20, 30, and 40 μg/ mL Xyl for 0, 5, 10, 15, 20, 25, 30, 45, 60, 90, and 120 min. Variations of amino acid neurotransmitters (AANTs), Nitric oxide-Cyclic GMP (NO-cGMP) signaling pathway, and ATPase were evaluated. Results: Xyl decreased the levels of cGMP and NO in nerve cells. Furthermore, Xyl affected the AANT content and Na+-K+-ATPase and Ca2+-Mg2+-ATPase activity in nerve cells. These findings suggested that Xyl inhibited the NO-cGMP signaling pathway in nerve cells in vitro. Conclusions: This study provided new evidence that the anesthetic and analgesic effects of Xyl are related to the inhibition of the NO-cGMP signaling pathway.

Estrogen promotes the onset and development of idiopathic scoliosis via disproportionate endochondral ossification of the anterior and posterior column in a bipedal rat model

Shuhui Zheng,Hang Zhou,Bo Gao,Yongyong Li,Zhiheng Liao,Taifeng Zhou,Chengjie Lian,Zizhao Wu,Deying Su,Tingting Wang,Peiqiang Su,Caixia Xu 생화학분자생물학회 2018 Experimental and molecular medicine Vol.50 No.-

This study aimed to verify the effects of estrogen on the onset and development of adolescent idiopathic scoliosis and the mechanisms associated with these effects by constructing a pubescent bipedal rat model. Experiments were conducted to investigate whether scoliosis progression was prevented by a Triptorelin treatment. One hundred twenty bipedal rats were divided into female, OVX (ovariectomy), OVX + E2, Triptorelin, sham, and male groups. According to a spinal radiographic analysis, the scoliosis rates and curve severity of the female and OVX + E2 groups were higher than those in the OVX, Triptorelin, and male groups. The measurements obtained from the sagittal plane of thoracic vertebrae CT confirmed a relatively slower growth of the anterior elements and a faster growth of the posterior elements between T11 and T13 in the female and OVX + E2 groups than in the OVX and Triptorelin groups. Histomorphometry and immunohistochemistry revealed a significantly longer hypertrophic zone of the vertebral cartilage growth plates that expressed more type X collagen and less type II collagen in the OVX and Triptorelin groups than in the female and OVX + E2 groups. Ki67 immunostaining confirmed an increase in the proliferation of vertebral growth plate chondrocytes in the OVX group compared with the female and OVX + E2 groups. In conclusion, estrogen obviously increased the incidence of scoliosis and curve severity in pubescent bipedal rats. The underlying mechanism may be a loss of coupling of the endochondral ossification between the anterior and posterior columns. Triptorelin decreased the incidence of scoliosis and curve magnitudes in bipedal female rats. Introduction Adolescent