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Proteomic Analysis on Exosomes Derived from Patients, Sera Infected with Echinococcus granulosus

  • Wang, Wen;Zhou, Xiaojing;Cui, Fang;Shi, Chunli;Wang, Yulan;Men, Yanfei;Zhao, Wei;Zhao, Jiaqing
    • Parasites, Hosts and Diseases
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    • v.57 no.5
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    • pp.489-497
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    • 2019
  • Cystic echinococcosis (CE), a zoonotic disease caused by Echinococcus granulosus at the larval stage, predominantly develops in the liver and lungs of intermediate hosts and eventually results in organ malfunction or even death. The interaction between E. granulosus and human body is incompletely understood. Exosomes are nanosized particles ubiquitously present in human body fluids. Exosomes carry biomolecules that facilitate communication between cells. To the best of our knowledge, the role of exosomes in patients with CE is not reported. Here, we isolated exosomes from the sera of patients with CE (CE-exo) and healthy donors and subjected them to liquid chromatography-tandem mass spectrometry analysis. Proteomic analysis identified 49 proteins specifically expressed in CE-exo, including 4 proteins of parasitic origin. The most valuable parasitic proteins included tubulin alpha-1C chain and histone H4. And 8 proteins were differentially regulated in CE-exo (fold change>1.5), as analyzed with bioinformatic methods such as annotation and functional enrichment analyses. These findings may improve our understanding about the interaction between E. granulosus and human body, and may contribute to the diagnosis and prevention of CE.

Underlying mechanisms of phosphodiesterase 10A and glutamate-ammonia ligase genes that regulate inosine monophosphate deposition and thereby affect muscle tenderness in Jingyuan chickens

  • Wang, Weizhen;Zhang, Juan;Hu, Honghong;Yu, Baojun;He, Jintong;Yao, Tingting;Gu, Yaling;Cai, Zhengyun;Xin, Guosheng
    • Animal Bioscience
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    • v.35 no.11
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    • pp.1771-1786
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    • 2022
  • Objective: Inosine monophosphate (IMP) is a key factor that imparts of meat flavor. Differences in the IMP content in the muscles were evaluated to improve chicken meat quality. Methods: For this study, the IMP content was detected by high performance liquid chromatography. The gene expression profiles of Jingyuan chickens with different feeding patterns and different sexes were analyzed by RNA-sequencing (RNA-seq). Results: Breast muscle IMP content in free-range chickens was extremely significantly higher than that of caged chickens (p<0.01). Breast muscle IMP content in hens was also higher than that of cocks, but the difference was not significant. Correlation analysis showed that the breast muscle IMP content in caged hens and cocks was negatively correlated with the shear force, and the breast muscle IMP content in free-range hens was significantly negatively correlated with the shear force (p<0.05). The two key genes associated with IMP synthesis in chickens with different feeding patterns were glutamate-ammonia ligase (GLUL) and phosphodiesterase 10A (PDE10A). Bioinformatics analysis revealed that the GLUL and PDE10A genes are involved in glutamine biosynthesis and purine salvage pathways respectively. In addition, GLUL expression was positively correlated with the IMP content in caged and free-range chickens, and PDE10A expression was significantly positively correlated with the IMP content in caged and free-range chickens (p<0.05). Conclusion: These findings will facilitate the comprehension of the deposition of IMP in the muscles and thereby aid the process of selection and breeding of good quality local chickens.

Lycium barbarum Polysaccharide Inhibits Lipid Oxidation and Protein Degradation in Tan Sheep Meatballs during Frozen Storage

  • Yu, Jiangyong;Guo, Mei;Liu, Guishan;Zhang, Jingjing;Fan, Naiyun;Li, Xiaorui;Sun, Yourui;Yuan, Jiangtao;Huang, Rui
    • Food Science of Animal Resources
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    • v.42 no.4
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    • pp.580-592
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    • 2022
  • The aim of the present study was to evaluate the effectiveness of Lycium barbarum polysaccharide (LBP) on lipid oxidation and protein degradation in Tan sheep meatballs during the frozen period. The meatballs were treated with LBP at 0.01%, 0.02%, and 0.03% and stored at -18±1℃ for 0, 3, 6, 9, and 12 weeks. The effects of LBP treatment were investigated using the contents of total volatile basic nitrogen (TVB-N), texture profile (TP), thiobarbituric acid reactive substances (TBARS), colour, and pH values, compared with 0.02% butylated hydroxytoluene treatment and the blank control. The results showed that LBP treatment significantly decreased TBARS content compared with the control, which confirmed LBP to be a highly effective component in preventing lipid oxidation of Tan sheep meatballs during frozen storage, and protein degradation in Tan sheep meatballs had a significant inhibition effect because of TVB-N value reduction. In addition, the colour, TP and pH values of meatballs treated with LBP were improved dramatically. To further determine the quality changes of the blank control and all treated groups during storage, the comprehensive score evaluation equation based on principal component analysis was obtained: Y=0.51632Y1+0.29589Y2 (cumulative contribution rate=81.221%), and the 0.02% LBP-treated group had a higher comprehensive score than the other groups, and the quality of LBP-treated meatballs was better as well. In summary, LBP may reduce or inhibit lipid oxidation and protein degradation, and enhance overall quality and shelf-life in prepared meat products.

Optimized study of an in vitro 3D culture of preantral follicles in mice

  • Hehe Ren;Yingxin Zhang;Yanping Zhang;Yikai Qiu;Qing Chang;Xiaoli Yu;Xiuying Pei
    • Journal of Veterinary Science
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    • v.24 no.1
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    • pp.4.1-4.16
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    • 2023
  • Background: In vitro culture of preantral follicles is a promising technology for fertility preservation. Objectives: This study aims to investigate an optimized three-dimensional (3D) fetal bovine serum (FBS)-free preantral follicle culture system having a simple and easy operation. Methods: The isolated follicles from mouse ovaries were randomly divided in an ultra-low attachment 96-well plates supplement with FBS or bovine serum albumin (BSA) culture or encapsulated with an alginate supplement with FBS or BSA culture. Meanwhile, estradiol (E2) concentration was assessed through enzyme-linked immunosorbent assay of culture supernatants. The diameter of follicular growth was measured, and the lumen of the follicle was photographed. Spindle microtubules of oocytes were detected via immunofluorescence. The ability of oocytes to fertilize was assessed using in vitro fertilization. Results: The diameters were larger for the growing secondary follicles cultured in ultra-low attachment 96-well plates than in the alginate gel on days 6, 8, and 10 (p < 0.05). Meanwhile, the E2 concentration in the BSA-supplemented medium was significantly higher in the alginate gel than in the other three groups on days 6 and 8 (p < 0.05), and the oocytes in the FBS-free system could complete meiosis and fertilization in vitro. Conclusions: The present study furnishes insights into the mature oocytes obtained from the 3D culture of the preantral follicle by using ultra-low attachment 96-well plate with an FBS-free system in vitro and supports the clinical practices to achieve competent, mature oocytes for in vitro fertilization.

Alfalfa xenomiR-162 targets G protein subunit gamma 11 to regulate milk protein synthesis in bovine mammary epithelial cells

  • Guizhi Meng;Hongjuan Duan;Jingying Jia;Baobao Liu;Yun Ma;Xiaoyan Cai
    • Animal Bioscience
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    • v.37 no.3
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    • pp.509-521
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    • 2024
  • Objective: It was shown that microRNAs (miRNAs) play an important role in milk protein synthesis. However, the post-transcriptional regulation of casein expression by exogenous miRNA (xeno-miRNAs) in ruminants remains unclear. This study explores the regulatory roles of alfalfa xeno-miR162 on casein synthesis in bovine mammary epithelial cells (bMECs). Methods: The effects of alfalfa xenomiR-162 and G protein subunit gamma 11 (GNG11) on proliferation and milk protein metabolism of bMECs were detected by 5-Ethynyl-2'-Deoxyuridine (EdU) staining, flow cytometry, cell counting kit-8 (CCK-8), enzyme-linked immunosorbent assay, quantitative real-time polymerase chain reaction (qRT-PCR), and Western blot. Dual-luciferase reporter assay was used to verify the targeting relationship between GNG11 and xenomiR-162. Results: Results showed that over-expression of xenomiR-162 inhibited cell proliferation but promoted apoptosis, which also up-regulated the expression of several casein coding genes, including CSN1S1, CSN1S2, and CSN3, while decreasing the expression of CSN2. Furthermore, the targeting relationship between GNG11 and xenomiR-162 was determined, and it was confirmed that GNG11 silencing also inhibited cell proliferation but promoted apoptosis and reduced the expression of casein coding genes and genes related to the mammalian target of rapamycin (mTOR) pathway. Conclusion: Alfalfa xenomiR-162 appears to regulate bMECs proliferation and milk protein synthesis via GNG11 in the mTOR pathway, suggesting that this xeno-miRNA could be harnessed to modulate CSN3 expression in dairy cows, and increase κ-casein contents in milk.

Upregulation of Fas in epithelial ovarian cancer reverses the development of resistance to Cisplatin

  • Fan, Yang;Wang, Long;Han, Xuechuan;Liu, Xueqin;Ma, Hongyun;Ding, Yonghui
    • BMB Reports
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    • v.48 no.1
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    • pp.30-35
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    • 2015
  • This study was to investigate the role of Fas in the development of Cisplatin-resistant ovarian cancer. On the cellular level, Fas expression was significantly reduced in Cisplatin resistant A2780 (A2780/CP) cells compared with A2780 cells. Fas silence with siRNA would promote tumor cell lines proliferation, facilitate tumor cell cycle transition of G1/S, prevent cell apoptosis, and promote cell migration. Expression of drug resistance gene was negatively correlated to Fas. In nude mice metastasis model of human ovarian carcinoma by subcutaneous transplantation, after Ad-Fas injected intratumorly, we found that upregulation of Fas could inhibit transplantation tumor tissue growth and reduce the expression of drug resistance gene. Our results indicated that upregulation of Fas in epithelial ovarian cancer reversed the development of resistance to Cisplatin. In conclusion, our findings suggested that Fas might act as a promising therapeutic target for improvement of the sensibility to Cisplatin in ovarian cancer.

The study of blood transcriptome profiles in Holstein cows with miscarriage during peri-implantation

  • Zhao, Guoli;Li, Yanyan;Kang, Xiaolong;Huang, Liang;Li, Peng;Zhou, Jinghang;Shi, Yuangang
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.1
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    • pp.38-48
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    • 2019
  • Objective: In this study, the transcriptome profile of cow experiencing miscarriage during peri-implantation was investigated. Methods: Total transcriptomes were checked by RNA sequencing, and the analyzed by bioinformatics methods, the differentially expressed genes (DEGs) were analysed with hierarchical clustering and Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis. Results: The results suggested that serum progesterone levels were significantly decreased in cows that miscarried as compared to the pregnant cows at 18, 21, 33, 39, and 51 days after artificial insemination. The RNA sequencing results suggested that 32, 176, 5, 10, and 2 DEGs were identified in the pregnant cows and miscarried cows at 18, 21, 33, 39, and 51 d after artificial insemination. And 15, 101, 1, 2, and 2 DEGs were upregulated, and 17, 74, 4, and 8 DEGs were downregulated in the cows in the pregnant and miscarriage groups, respectively at 18, 21, 33, and 39, but no gene was downregulated at 51 d after artificial insemination. These DEGs were distributed to 13, 20, 3, 6, and 20 pathways, and some pathway essential for pregnancy, such as cell adhesion molecules, tumor necrosis factor signaling pathway and PI3K-Akt signaling pathway. Conclusion: This analysis has identified several genes and related pathways crucial for pregnancy and miscarriage in cows, as well as these genes supply molecular markers to predict the miscarriage in cows.

In silico genome wide identification and expression analysis of the WUSCHEL-related homeobox gene family in Medicago sativa

  • Yang, Tianhui;Gao, Ting;Wang, Chuang;Wang, Xiaochun;Chen, Caijin;Tian, Mei;Yang, Weidi
    • Genomics & Informatics
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    • v.20 no.2
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    • pp.19.1-19.15
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    • 2022
  • Alfalfa (Medicago sativa) is an important food and feed crop which rich in mineral sources. The WUSCHEL-related homeobox (WOX) gene family plays important roles in plant development and identification of putative gene families, their structure, and potential functions is a primary step for not only understanding the genetic mechanisms behind various biological process but also for genetic improvement. A variety of computational tools, including MAFFT, HMMER, hidden Markov models, Pfam, SMART, MEGA, ProtTest, BLASTn, and BRAD, among others, were used. We identified 34 MsWOX genes based on a systematic analysis of the alfalfa plant genome spread in eight chromosomes. This is an expansion of the gene family which we attribute to observed chromosomal duplications. Sequence alignment analysis revealed 61 conserved proteins containing a homeodomain. Phylogenetic study sung reveal five evolutionary clades with 15 motif distributions. Gene structure analysis reveals various exon, intron, and untranslated structures which are consistent in genes from similar clades. Functional analysis prediction of promoter regions reveals various transcription binding sites containing key growth, development, and stress-responsive transcription factor families such as MYB, ERF, AP2, and NAC which are spread across the genes. Most of the genes are predicted to be in the nucleus. Also, there are duplication events in some genes which explain the expansion of the family. The present research provides a clue on the potential roles of MsWOX family genes that will be useful for further understanding their functional roles in alfalfa plants.

A Study on the Motivation and Choice Attributes of Visiting Wineries in Ningxia, China (중국 닝샤 와이너리 방문동기와 선택속성에 관한 연구)

  • Li, Shu-Xian;Woo, Won-Seok;Hwang, Jae-Hyun
    • Korean Journal of Organic Agriculture
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    • v.32 no.1
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    • pp.39-54
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    • 2024
  • This study aimed to elucidate the relationship between the selection attributes and visit motivations of Ningxia wineries and tourist satisfaction and intention to revisit. Utilizing multiple linear regression and simple linear regression models, the study quantitatively analyzed tourists' selection attributes and visit motivations for Ningxia wineries. Factor analysis categorized 12 visit motivation variables into four factors: 'Sensation', 'Educational', 'Companion-Friendly', and 'Experiential'. Additionally, 17 selection attribute variables were classified into four factors: 'Service Quality', 'Environmental', 'Facility', and 'Economic'. Through analyzing the impact of visit motivations on satisfaction and revisit intentions, we have identified the pivotal factors as 'Wine Cultural Education', 'Enhanced Companion-Friendliness', and 'Wine Cultural Experience'. In the analysis of the relationship between selection attributes and revisit intentions, crucial elements involve 'Service Quality provided by the winery"'and 'Environmental of the winery'. Conversely, key influencing revisit intentions include 'Environmental of the winery' and 'Costs and Pricing associated with winery visits'. To ensure the sustainable development of the Ningxia winery industry and promote the enduring growth of rural economies, wineries should place greater emphasis on cultivating wine cultural experiences, artisanal activities, and other project endeavors.

LINC00174 Facilitates Proliferation and Migration of Colorectal Cancer Cells via MiR-3127-5p/ E2F7 Axis

  • Ma, Yuhong;Li, Yuzhen;Tang, Yuanyuan;Tang, Ning;Wang, Dengke;Li, Xiaofei
    • Journal of Microbiology and Biotechnology
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    • v.31 no.8
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    • pp.1098-1108
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    • 2021
  • The literature indicates that LINC00174 promotes the growth of colorectal cancer (CRC) cells, but its research needs to be enriched. We tried to explore the function and mechanism of LINC00174 in CRC cell proliferation and migration. Bioinformatics analysis predicted the binding relationship and expressions of lncRNA, miRNA and mRNA. Clinical study analyzes the relationship between LINC00174 and clinical data characteristics of CRC patients. The expressions of LINC00174, miR-3127-5p and E2F7 were verified by RT-qPCR, and the combination of the two was verified by dual luciferase analysis and RNA immunoprecipitation as needed. Western blot was used to detect the expression of EMT-related protein and E2F7 protein. Functional experiments were used to evaluate the function of the target gene on CRC cells. LINC00174 was up-regulated in CRC clinical samples and cells and was related to the clinical characteristics of CRC patients. High-expression of LINC00174, contrary to the effect of siLINC00174, promoted cell viability, proliferation, migration and invasion, up-regulated the expressions of N-Cadherin, Vimentin, E2F7, and inhibited the expression of E-Cadherin. MiR-3127-5p was one of the targeted miRNAs of LINC00174 and was down-regulated in CRC samples. In addition, miR-3127-5p mimic partially reversed the malignant phenotype of CRC cells induced by LINC00174. Besides, E2F7 was a target gene of miR-3127-5p, and LINC00174 repressed miR-3127-5p to regulate E2F7. Our research reveals that LINC00174 affected the biological characteristics of CRC cells through regulated miR-3127-5p/ E2F7 axis.