Graphene Oxide Nanosheet-Composited Poly(N-isopropylacrylamide) Hydrogel for Cell Sheet Recovery

Yongqing Xia,Han Wu,Dachao Tang,Shuai Gao,Binghe Chen,Zhujun Zeng,Shengjie Wang,Meiwen Cao,Dongxiang Li 한국고분자학회 2019 Macromolecular Research Vol.27 No.7

Cell sheet engineering technique has been applied to treat various tissues without the use of a traditional scaffold. To date, methods for the cell sheet harvesting depend mostly on grafted poly(N-isopropylacrylamide) (pNIPAAm) thin layers, while the native pNIPAAm hydrogel, which possibly presents the easiest way to prepare thermo-responsive materials, is not suitable for the cell sheet harvesting due to its low cell attachment. In this study, the graphene oxide (GO) nanosheet was utilized as an additive to enhance the bio-compatibility of the pNIPAAm hydrogel. Different concentrations of GO nanosheets were added to prepare GO/pNIPAAm composite hydrogels through the in-situ free radical polymerization with polyethylene glycol dimethacrylate (PEGDA) as a cross-linker. The results indicated that the physical properties of the composite hydrogels had little difference with that of the native pNIPAAm hydrogel. However, the cell attachment, proliferation and detachment behaviors on the composite hydrogel surface were greatly enhanced. Monkey fibroblast COS7 cells attached and proliferated better on the GO/pNIPAAm composite hydrogel, while intact COS7 cell sheets could be harvested from the composite hydrogels by simply lowering the temperature. In contrast, the cells appeared as clusters on the native pNIPAAm hydrogel. Furthermore, when HeLa and COS7 cells were seeded successively onto the micropatterned GO/pNIPAAm hydrogel, there could be the formation of a patterned HeLa/COS7 cell layer. The geometrically patterned GO/pNIPAAm hydrogel may provide an easy-to-prepare material for releasing patterned cell sheets compared to the specific cell-adhesive proteins reported to make patterned cell layers.

Comparative analysis of liver transcriptome reveals adaptive responses to hypoxia environmental condition in Tibetan chicken

Yongqing Cao,Tao Zeng,Wei Han,Xueying Ma,Tiantian Gu,Li Chen,Yong Tian,Wenwu Xu,Jianmei Yin,Guohui Li,Lizhi Lu,Shuangbao Gun Asian Australasian Association of Animal Productio 2024 Animal Bioscience Vol.37 No.1

Objective: Tibetan chickens, which have unique adaptations to extreme high-altitude environments, exhibit phenotypic and physiological characteristics that are distinct from those of lowland chickens. However, the mechanisms underlying hypoxic adaptation in the liver of chickens remain unknown. Methods: RNA-sequencing (RNA-Seq) technology was used to assess the differentially expressed genes (DEGs) involved in hypoxia adaptation in highland chickens (native Tibetan chicken [HT]) and lowland chickens (Langshan chicken [LS], Beijing You chicken [BJ], Qingyuan Partridge chicken [QY], and Chahua chicken [CH]). Results: A total of 352 co-DEGs were specifically screened between HT and four native lowland chicken breeds. Gene ontology and Kyoto encyclopedia of genes and genomes enrichment analyses indicated that these co-DEGs were widely involved in lipid metabolism processes, such as the peroxisome proliferator-activated receptors (PPAR) signaling pathway, fatty acid degradation, fatty acid metabolism and fatty acid biosynthesis. To further determine the relationship from the 352 co-DEGs, protein-protein interaction network was carried out and identified eight genes (ACSL1, CPT1A, ACOX1, PPARC1A, SCD, ACSBG2, ACACA, and FASN) as the potential regulating genes that are responsible for the altitude difference between the HT and other four lowland chicken breeds. Conclusion: This study provides novel insights into the molecular mechanisms regulating hypoxia adaptation via lipid metabolism in Tibetan chickens and other highland animals.

Effects of TLR9/NF-κB on oxidative stress and inflammation in IPEC-J2 cells

Ma Lixia,Geng Jinhong,Chen Wei,Qin Ming,Wang Lixue,Zeng Yongqing 한국유전학회 2022 Genes & Genomics Vol.44 No.10

Background: Oxidative stress is one of the most important factors affecting large-scale breeding, especially the performance of pigs. Oxidative stress plays a role by affecting various genes in pigs, which can cause serious body damage, functional degradation and reduce production performance. Objective: The purpose of this study was to investigate the effect of Toll like receptor 9 (TLR9) pathway on IPEC-J2 cells under oxidative stress and to provide reference for the growth development of Dapulian pigs. Methods: In this study, Diquat was used as a source of oxidative stress to study the effects on Dapulian pigs by detecting relevant indicators. Then the IPEC-J2 cells were selected to verify the TLR9 signaling pathway in oxidative stress. Results: Compared with the control group, superoxide dismutase (SOD) in experimental group decreased significantly, malondialdehyde (MDA) was significantly increased, accompanied by inflammatory reaction, and inflammatory factors were significantly increased in the experimental group. Oxidative stress model was constructed by H2O2 incubating IPEC-J2 cells. The interference and overexpression vectors of TLR9 and myeloid differentiation primary response protein 88 (MyD88) were constructed to detect the activity of antioxidant enzymes and related proteins. The results showed that overexpression of TLR9 enhanced the activity of antioxidant enzymes, decreased the secretion of inflammatory factors, and decreased the activity of MDA,reactive oxygen species (ROS); the results were opposite after TLR9 interference. This study also showed that H2O2 can activate the nuclear factor-κB (NF-κB) pathway and promote the translocation of NF-κB into the nucleus. After co-transfection with TLR9 and MyD88, the results showed that TLR9 regulated the expression of NF-κB through MyD88. Conclusion: The study showed that TLR9 pathway had a significant positive effect on antioxidant.

The role of long noncoding RNAs in livestock adipose tissue deposition - A review

Wang, Lixue,Xie, Yuhuai,Chen, Wei,Zhang, Yu,Zeng, Yongqing Asian Australasian Association of Animal Productio 2021 Animal Bioscience Vol.34 No.7

With the development of sequencing technology, numerous, long noncoding RNAs (lncRNAs) have been discovered and annotated. Increasing evidence has shown that lncRNAs play an essential role in regulating many biological and pathological processes, especially in cancer. However, there have been few studies on the roles of lncRNAs in livestock production. In animal products, meat quality and lean percentage are vital economic traits closely related to adipose tissue deposition. However, adipose tissue accumulation is also a pivotal contributor to obesity, diabetes, atherosclerosis, and many other diseases, as demonstrated by human studies. In livestock production, the mechanism by which lncRNAs regulate adipose tissue deposition is still unclear. In addition, the phenomenon that different animal species have different adipose tissue accumulation abilities is not well understood. In this review, we summarize the characteristics of lncRNAs and their four functional archetypes and review the current knowledge about lncRNA functions in adipose tissue deposition in livestock species. This review could provide theoretical significance to explore the functional mechanisms of lncRNAs in adipose tissue accumulation in animals.

Identification and functional prediction of long non-coding RNAs related to skeletal muscle development in Duroc pigs

Ma Lixia,Qin Ming,Zhang Yulun,Xue Hui,Li Shiyin,Chen Wei,Zeng Yongqing 아세아·태평양축산학회 2022 Animal Bioscience Vol.35 No.10

Objective: The growth of pigs involves multiple regulatory mechanisms, and modern molecular breeding techniques can be used to understand the skeletal muscle growth and development to promote the selection process of pigs. This study aims to explore candidate lncRNAs and mRNAs related to skeletal muscle growth and development among Duroc pigs with different average daily gain (ADG). Methods: A total of 8 pigs were selected and divided into two groups: H group (high-ADG) and L group (low-ADG). And followed by whole transcriptome sequencing to identify differentially expressed (DE) lncRNAs and mRNAs. Results: In RNA-seq, 703 DE mRNAs (263 up-regulated and 440 down-regulated) and 74 DE lncRNAs (45 up-regulated and 29 down-regulated) were identified. In addition, 1,418 Transcription factors (TFs) were found. Compared with mRNAs, lncRNAs had fewer exons, shorter transcript length and open reading frame length. DE mRNAs and DE lncRNAs can form 417 lncRNA-mRNA pairs (antisense, cis and trans). DE mRNAs and target genes of lncRNAs were enriched in cellular processes, biological regulation, and regulation of biological processes. In addition, quantitative trait locus (QTL) analysis was used to detect the functions of DE mRNAs and lncRNAs, the most of DE mRNAs and target genes of lncRNAs were enriched in QTLs related to growth traits and skeletal muscle development. In single-nucleotide polymorphism/insertion-deletion (SNP/INDEL) analysis, 1,081,182 SNP and 131,721 INDEL were found, and transition was more than transversion. Over 60% of percentage were skipped exon events among alternative splicing events. Conclusion: The results showed that different ADG among Duroc pigs with the same diet maybe due to the DE mRNAs and DE lncRNAs related to skeletal muscle growth and development. Objective: The growth of pigs involves multiple regulatory mechanisms, and modern molecular breeding techniques can be used to understand the skeletal muscle growth and development to promote the selection process of pigs. This study aims to explore candidate lncRNAs and mRNAs related to skeletal muscle growth and development among Duroc pigs with different average daily gain (ADG).Methods: A total of 8 pigs were selected and divided into two groups: H group (high-ADG) and L group (low-ADG). And followed by whole transcriptome sequencing to identify differentially expressed (DE) lncRNAs and mRNAs.Results: In RNA-seq, 703 DE mRNAs (263 up-regulated and 440 down-regulated) and 74 DE lncRNAs (45 up-regulated and 29 down-regulated) were identified. In addition, 1,418 Transcription factors (TFs) were found. Compared with mRNAs, lncRNAs had fewer exons, shorter transcript length and open reading frame length. DE mRNAs and DE lncRNAs can form 417 lncRNA-mRNA pairs (antisense, cis and trans). DE mRNAs and target genes of lncRNAs were enriched in cellular processes, biological regulation, and regulation of biological processes. In addition, quantitative trait locus (QTL) analysis was used to detect the functions of DE mRNAs and lncRNAs, the most of DE mRNAs and target genes of lncRNAs were enriched in QTLs related to growth traits and skeletal muscle development. In single-nucleotide polymorphism/insertion-deletion (SNP/INDEL) analysis, 1,081,182 SNP and 131,721 INDEL were found, and transition was more than transversion. Over 60% of percentage were skipped exon events among alternative splicing events.Conclusion: The results showed that different ADG among Duroc pigs with the same diet maybe due to the DE mRNAs and DE lncRNAs related to skeletal muscle growth and development.

Identification and functional prediction of long non-coding RNAs related to oxidative stress in the jejunum of piglets

Li Jinbao,Zhang Jianmin,Jin Xinlin,Li Shiyin,Du Yingbin,Zeng Yongqing,Wang Jin,Chen Wei 아세아·태평양축산학회 2024 Animal Bioscience Vol.37 No.2

Objective: Oxidative stress (OS) is a pathological process arising from the excessive production of free radicals in the body. It has the potential to alter animal gene expression and cause damage to the jejunum. However, there have been few reports of changes in the expression of long noncoding RNAs (lncRNAs) in the jejunum in piglets under OS. The purpose of this research was to examine how lncRNAs in piglet jejunum change under OS. Methods: The abdominal cavities of piglets were injected with diquat (DQ) to produce OS. Raw reads were downloaded from the SRA database. RNA-seq was utilized to study the expression of lncRNAs in piglets under OS. Additionally, six randomly selected lncRNAs were verified using quantitative real-time polymerase chain reaction (qRT‒PCR) to examine the mechanism of oxidative damage. Results: A total of 79 lncRNAs were differentially expressed (DE) in the treatment group compared to the negative control group. The target genes of DE lncRNAs were enriched in gene ontology (GO) terms and Kyoto encyclopedia of genes and genomes (KEGG) signaling pathways. Chemical carcinogenesis-reactive oxygen species, the Foxo signaling pathway, colorectal cancer, and the AMPK signaling pathway were all linked to OS. Conclusion: Our results demonstrated that DQ-induced OS causes differential expression of lncRNAs, laying the groundwork for future research into the processes involved in the jejunum’s response to OS.

Identification and functional prediction of long noncoding RNAs related to intramuscular fat content in Laiwu pigs

Wang Lixue,Xie Yuhuai,Chen Wei,Zhang Yu,Zeng Yongqing 아세아·태평양축산학회 2022 Animal Bioscience Vol.35 No.1

Objective: Intramuscular fat (IMF) is a critical economic indicator of pork quality. Studies on IMF among different pig breeds have been performed via high-throughput sequencing, but comparisons within the same pig breed remain unreported. Methods: This study was performed to explore the gene profile and identify candidate long noncoding RNA (lncRNAs) and mRNAs associated with IMF deposition among Laiwu pigs with different IMF contents. Based on the longissimus dorsi muscle IMF content, eight pigs from the same breed and management were selected and divided into two groups: a high IMF (>12%, H) and low IMF group (<5%, L). Whole-transcriptome sequencing was performed to explore the differentially expressed (DE) genes between these two groups. Results: The IMF content varied greatly among Laiwu pig individuals (2.17% to 13.93%). Seventeen DE lncRNAs (11 upregulated and 6 downregulated) and 180 mRNAs (112 upregulated and 68 downregulated) were found. Gene Ontology analysis indicated that the following biological processes played an important role in IMF deposition: fatty acid and lipid biosynthetic processes; the extracellular signal-regulated kinase cascade; and white fat cell differentiation. In addition, the peroxisome proliferator-activated receptor, phosphatidylinositol- 3-kinase-protein kinase B, and mammalian target of rapamycin pathways were enriched in the pathway analysis. Intersection analysis of the target genes of DE lncRNAs and mRNAs revealed seven candidate genes associated with IMF accumulation. Five DE lncRNAs and 20 DE mRNAs based on the pig quantitative trait locus database were identified and shown to be related to fat deposition. The expression of five DE lncRNAs and mRNAs was verified by quantitative real time polymerase chain reaction (qRT-PCR). The results of qRT-PCR and RNA-sequencing were consistent. Conclusion: These results demonstrated that the different IMF contents among pig individuals may be due to the DE lncRNAs and mRNAs associated with lipid droplets and fat deposition. Objective: Intramuscular fat (IMF) is a critical economic indicator of pork quality. Studies on IMF among different pig breeds have been performed via high-throughput sequencing, but comparisons within the same pig breed remain unreported.Methods: This study was performed to explore the gene profile and identify candidate long noncoding RNA (lncRNAs) and mRNAs associated with IMF deposition among Laiwu pigs with different IMF contents. Based on the longissimus dorsi muscle IMF content, eight pigs from the same breed and management were selected and divided into two groups: a high IMF (>12%, H) and low IMF group (<5%, L). Whole-transcriptome sequencing was performed to explore the differentially expressed (DE) genes between these two groups.Results: The IMF content varied greatly among Laiwu pig individuals (2.17% to 13.93%). Seventeen DE lncRNAs (11 upregulated and 6 downregulated) and 180 mRNAs (112 upregulated and 68 downregulated) were found. Gene Ontology analysis indicated that the following biological processes played an important role in IMF deposition: fatty acid and lipid biosynthetic processes; the extracellular signal-regulated kinase cascade; and white fat cell differentiation. In addition, the peroxisome proliferator-activated receptor, phosphatidylinositol-3-kinase-protein kinase B, and mammalian target of rapamycin pathways were enriched in the pathway analysis. Intersection analysis of the target genes of DE lncRNAs and mRNAs revealed seven candidate genes associated with IMF accumulation. Five DE lncRNAs and 20 DE mRNAs based on the pig quantitative trait locus database were identified and shown to be related to fat deposition. The expression of five DE lncRNAs and mRNAs was verified by quantitative real time polymerase chain reaction (qRT-PCR). The results of qRT-PCR and RNA-sequencing were consistent.Conclusion: These results demonstrated that the different IMF contents among pig individuals may be due to the DE lncRNAs and mRNAs associated with lipid droplets and fat deposition.