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Article: The spatio-temporal expression analysis of parathyroid hormone like hormone gene provide a new insight for bone growth of the antler tip tissue in sika deer.

Xing, Haihua / Han, Ruobing / Wang, Qianghui / Sun, Zihui / Li, Heping

Animal bioscience

2024

Abstract: Objective: Parathyroid hormone like hormone (PTHLH), as an essential factor for bone growth, is involved in a variety of physiological processes. The aim of this study was to explore the role of PTHLH gene in the growth of antlers.: Methods: The ... ...

Abstract	Objective: Parathyroid hormone like hormone (PTHLH), as an essential factor for bone growth, is involved in a variety of physiological processes. The aim of this study was to explore the role of PTHLH gene in the growth of antlers. Methods: The coding sequence (CDS) of PTHLH gene cDNA was obtained by cloning in sika deer (Cervus nippon), and the bioinformatics was analyzed. The quantitative real-time polymerase chain reaction (qRT-PCR) was used to analyze the differences expression of PTHLH mRNA in different tissues of the antler tip at different growth periods (early period, EP; middle period, MP; late period, LP). Results: The CDS of PTHLH gene was 534bp in length and encoded 177 amino acids. Predictive analysis results revealed that the PTHLH protein was a hydrophilic protein without transmembrane structure, with its secondary structure consisting mainly of random coil. The PTHLH protein of sika deer had the identity of 98.31%, 96.82%, 96.05% and 94.92% with Cervus canadensis, Bos mutus, Oryx dammah and Budorcas taxicolor, which were highly conserved among the artiodactyls. The qRT-PCR results showed that PTHLH mRNA had a unique spatio-temporal expression pattern in antlers. In the dermis, precartilage, and cartilage tissues, the expression of PTHLH mRNA was extremely significantly higher in MP than in EP, LP (p<0.01). In the mesenchyme tissue, the expression of PTHLH mRNA in MP was significantly higher than that of EP (p<0.05), but extremely significantly lower than that of LP (p<0.01). The expression of PTHLH mRNA in antler tip tissues at all growth periods had approximately the same trend, that is, from distal to basal, it was first down-regulated from the dermis to the mesenchyme and then continuously up-regulated to the cartilage tissue. Conclusion: PTHLH gene may promote the rapid growth of antler mainly through its extensive regulatory effect on the antler tip tissue.
Language	English
Publishing date	2024-02-28
Publishing country	Korea (South)
Document type	Journal Article
ISSN	2765-0189
ISSN	2765-0189
DOI	10.5713/ab.23.0421
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: DNA methylation pattern and mRNA expression of OPN promoter in sika deer antler tip tissues

Xing, Haihua / Zhang, Furui / Han, Ruobing / Li, Heping

Gene. 2023 June, v. 868 p.147382-

2023

Abstract: In order to explore the biological role of OPN gene during the growth of sika deer antler, the dermis, mesenchyme, precartilage and cartilage tissues of sika deer antler tip at the early period of the antler with a saddle-like appearance (30 days), the ... ...

Abstract	In order to explore the biological role of OPN gene during the growth of sika deer antler, the dermis, mesenchyme, precartilage and cartilage tissues of sika deer antler tip at the early period of the antler with a saddle-like appearance (30 days), the rapid growth period of the antler with two branches (60 days), and the final period of the antler with three branches (90 days) were analyzed. Bisulfite sequencing PCR (BSP) and quantitative real-time PCR (qRT-PCR) were used to explore the DNA promoter methylation and mRNA expression of OPN in sika deer antler from the perspective of space and time. The test results showed that: 1) The methylation rates of OPN promoter at the early, middle and late periods of dermis tissue were (40.48 ± 0.82)%, (40.00 ± 1.43)%, and (39.05 ± 0.82)%; The methylation rates in mesenchyme tissue were (37.62 ± 0.82)%, (34.76 ± 2.18)%, and (38.57 ± 1.43)%; The methylation rates in precartilage tissue were (36.67 ± 0.28)%, (29.52 ± 1.65)%, (28.10 ± 2.18)%; The methylation rates in cartilage tissue were (31.90 ± 1.65)%, (26.67 ± 1.65)%, (24.29 ± 1.43)%. 2) There are 7 CpG sites in the OPN promoter region, and the 3 CpG sites of −367 bp, −245 bp and −31 bp are all methylated to different level. 3) The methylation level of OPN in the dermis, mesenchyme, precartilage and cartilage tissues decreased in sequence at the same growth period. At the middle and late periods, the methylation level of the promoter region of the precartilage tissue was significantly different from that of the dermis and mesenchyme tissues (P < 0.05); At different growth periods, the methylation level of the promoter region of cartilage tissue was extremely significantly different from that of dermis and mesenchyme tissues (P < 0.01); In the same tissue, the methylation level of the promoter region at the middle period was down-regulated compared with the early period, and the methylation level of the promoter region at the early period and the middle period was extremely significantly different in the precartilage and cartilage (P < 0.01). 4) OPN mRNA is highly expressed in precartilage and cartilage tissues. 5) The methylation level of OPN promoter was negatively correlated with mRNA expression level. In summary, it is speculated that the OPN gene, which may be regulated by the DNA methylation level of the promoter, promotes the growth and development of deer antler mainly by regulating the growth of precartilage and cartilage tissues.
Keywords	Cervus nippon ; DNA ; DNA methylation ; bisulfites ; cartilage ; deer ; dermis ; gene expression ; genes ; growth and development ; promoter regions ; quantitative polymerase chain reaction ; space and time ; Antler ; OPN gene ; BSP technology ; mRNA expression ; AGC ; bp ; BSP ; CpG ; DEGs ; DEPs ; EP ; LP ; MP ; MSCs ; OPN ; PCR ; qRT-PCR ; RGD ; SPP1 ; TSS
Language	English
Dates of publication	2023-06
Publishing place	Elsevier B.V.
Document type	Article ; Online
Note	Use and reproduction
ZDB-ID	391792-7
ISSN	1879-0038 ; 0378-1119
ISSN (online)	1879-0038
ISSN	0378-1119
DOI	10.1016/j.gene.2023.147382
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Article ; Online: DNA methylation pattern and mRNA expression of OPN promoter in sika deer antler tip tissues.

Xing, Haihua / Zhang, Furui / Han, Ruobing / Li, Heping

Gene

2023 Volume 868, Page(s) 147382

Abstract	In order to explore the biological role of OPN gene during the growth of sika deer antler, the dermis, mesenchyme, precartilage and cartilage tissues of sika deer antler tip at the early period of the antler with a saddle-like appearance (30 days), the rapid growth period of the antler with two branches (60 days), and the final period of the antler with three branches (90 days) were analyzed. Bisulfite sequencing PCR (BSP) and quantitative real-time PCR (qRT-PCR) were used to explore the DNA promoter methylation and mRNA expression of OPN in sika deer antler from the perspective of space and time. The test results showed that: 1) The methylation rates of OPN promoter at the early, middle and late periods of dermis tissue were (40.48 ± 0.82)%, (40.00 ± 1.43)%, and (39.05 ± 0.82)%; The methylation rates in mesenchyme tissue were (37.62 ± 0.82)%, (34.76 ± 2.18)%, and (38.57 ± 1.43)%; The methylation rates in precartilage tissue were (36.67 ± 0.28)%, (29.52 ± 1.65)%, (28.10 ± 2.18)%; The methylation rates in cartilage tissue were (31.90 ± 1.65)%, (26.67 ± 1.65)%, (24.29 ± 1.43)%. 2) There are 7 CpG sites in the OPN promoter region, and the 3 CpG sites of -367 bp, -245 bp and -31 bp are all methylated to different level. 3) The methylation level of OPN in the dermis, mesenchyme, precartilage and cartilage tissues decreased in sequence at the same growth period. At the middle and late periods, the methylation level of the promoter region of the precartilage tissue was significantly different from that of the dermis and mesenchyme tissues (P < 0.05); At different growth periods, the methylation level of the promoter region of cartilage tissue was extremely significantly different from that of dermis and mesenchyme tissues (P < 0.01); In the same tissue, the methylation level of the promoter region at the middle period was down-regulated compared with the early period, and the methylation level of the promoter region at the early period and the middle period was extremely significantly different in the precartilage and cartilage (P < 0.01). 4) OPN mRNA is highly expressed in precartilage and cartilage tissues. 5) The methylation level of OPN promoter was negatively correlated with mRNA expression level. In summary, it is speculated that the OPN gene, which may be regulated by the DNA methylation level of the promoter, promotes the growth and development of deer antler mainly by regulating the growth of precartilage and cartilage tissues.
MeSH term(s)	Animals ; Deer/genetics ; DNA Methylation ; Antlers/physiology ; RNA, Messenger/metabolism ; Promoter Regions, Genetic ; Real-Time Polymerase Chain Reaction
Chemical Substances	RNA, Messenger
Language	English
Publishing date	2023-03-21
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	391792-7
ISSN	1879-0038 ; 0378-1119
ISSN (online)	1879-0038
ISSN	0378-1119
DOI	10.1016/j.gene.2023.147382
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Article: Assessment of genetic diversity of Cervus albirostris (white-lipped deer) by microsatellite markers

Han, Ruobing / Zhao, Hongnan / Li, Defa / Li, Heping

Animal biotechnology. 2022 Aug. 1, v. 33, no. 4

2022

Abstract: Cervus albirostris (white-lipped deer) is a unique plateau deer in China. In order to conserve the white-lipped deer population and maintain its sustainable development in Liaoyang, China, ten microsatellite markers was used to analyze the genetic ... ...

Abstract	Cervus albirostris (white-lipped deer) is a unique plateau deer in China. In order to conserve the white-lipped deer population and maintain its sustainable development in Liaoyang, China, ten microsatellite markers was used to analyze the genetic diversity of 42 white-lipped deer, an ex-situ conservation population from Yushu (Qinghai) to Liaoyang (Liaoning). The results showed that expected heterozygosity ranged from 0.399 to 0.825 with a mean of 0.6071, while observed heterozygosity varied from 0.081 to 0.692 with a mean of 0.4456. This study contributes to the study of the unique genetic resources in this ex-situ conservation population and provides a perspective for better implementation of ex-situ conservation of white-lipped deer population in Liaoyang, China.
Keywords	Cervus ; biotechnology ; deer ; ex situ conservation ; genetic variation ; heterozygosity ; microsatellite repeats ; sustainable development ; China
Language	English
Dates of publication	2022-0801
Size	p. 739-744.
Publishing place	Taylor & Francis
Document type	Article
ZDB-ID	2043243-4
ISSN	1532-2378 ; 1049-5398
ISSN (online)	1532-2378
ISSN	1049-5398
DOI	10.1080/10495398.2020.1832104
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Article ; Online: lncRNA Sequencing of Antler Mesenchymal Tissue Revealed that the Regulatory Network of Antler Cell Proliferation and Differentiation

Han, Ruobing / Han, Lei / Xia, Yanling / Guo, Mengya / Li, Heping

Animal Biotechnology. 2022 Dec. 12, v. 33, no. 7 p.1629-1638

2022

Abstract: Antlers have been widely studied due to their unique physiological characteristics, such as rapid growth, periodic shedding and regeneration. However, little is known about how antler growth is regulated by long non-coding RNA (lncRNA). The aim of the ... ...

Abstract	Antlers have been widely studied due to their unique physiological characteristics, such as rapid growth, periodic shedding and regeneration. However, little is known about how antler growth is regulated by long non-coding RNA (lncRNA). The aim of the present study was to identify the lncRNAs expression profile and explore the function of lncRNAs during the antler growth. Herein, RNA-sequencing technology (RNA-seq) was performed on the three growth periods (early developmental period: EP, middle developmental period: MP, later developmental period: LP) of male sika deer (Cervus nippon) antler, 16 differentially expressed lncRNAs (DE lncRNAs) and 11 DE lncRNAs were identified in EP vs MP and MP vs LP related to cell proliferation and cell differentiation, respectively. Finally, lncRNAs-mRNAs co-expression networks were constructed based on the identified DE lncRNAs and their potential trans-target genes. The result reveals that lncRNAs may play diverse roles in different periods of antler growth. It provides a novel perspective for revealing the molecular mechanism of antler growth.
Keywords	Cervus nippon ; animals ; biotechnology ; cell differentiation ; cell proliferation ; males ; non-coding RNA ; sequence analysis ; Antlers ; lncRNAs ; RNA-seq ; co-expression network
Language	English
Dates of publication	2022-1212
Size	p. 1629-1638.
Publishing place	Taylor & Francis
Document type	Article ; Online
ZDB-ID	2043243-4
ISSN	1532-2378 ; 1049-5398
ISSN (online)	1532-2378
ISSN	1049-5398
DOI	10.1080/10495398.2021.1924762
Database	NAL-Catalogue (AGRICOLA)

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Article: Whole Transcriptome Analysis of Mesenchyme Tissue in Sika Deer Antler Revealed the CeRNAs Regulatory Network Associated With Antler Development.

Han, Ruobing / Han, Lei / Wang, Shengnan / Li, Heping

Frontiers in genetics

2020 Volume 10, Page(s) 1403

Abstract: Deer antler is the only completely regenerable organ in mammals. During the rapid growth period, the antler proliferates even faster than cancerous tissue growth. However, the proliferation and development of antler have been in a stable and controllable ...

Abstract	Deer antler is the only completely regenerable organ in mammals. During the rapid growth period, the antler proliferates even faster than cancerous tissue growth. However, the proliferation and development of antler have been in a stable and controllable growth cycle. In this study, we analyzed the time series expression data of nine samples from mesenchyme layer in three male sika deer in the early period of the antler with a saddle-like appearance (30 days), the rapid growth period of the antler with two branches (60 days), and the final period of the antler with three branches (90 days). Whole Transcriptome sequencing results show that in the 30 d versus 60 d group, 1,464 genes, 85 long noncoding RNAs (lncRNAs), and 61 miRNAs were identified as differentially expressed; 1,748 genes, 138 lncRNAs, and 78 miRNAs were identified as differentially expressed in 30d versus 90d group; and 816 differentially expressed genes (DEGs), 49 differentially expressed lncRNAs (DE lncRNAs), and 24 differentially expressed miRNA (DE miRNAs) were identified in 60d versus 90d group. A total of 182 miRNA-mRNA interaction pairs and 89 miRNA-lncRNA interaction pairs were screened from DEGs, DE miRNAs, and DE lncRNAs to construct the ceRNA regulatory network (ceRNET). In summary, we identified candidate mRNAs, miRNAs and lncRNAs that regulate the development of antler tip. It may lay the foundation for further investigating the molecular mechanism of antler rapid growth and development.
Language	English
Publishing date	2020-02-18
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2606823-0
ISSN	1664-8021
ISSN	1664-8021
DOI	10.3389/fgene.2019.01403
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Article ; Online: Assessment of genetic diversity of

Han, Ruobing / Zhao, Hongnan / Li, Defa / Li, Heping

Animal biotechnology

2020 Volume 33, Issue 4, Page(s) 739–744

Abstract: ... Cervus ... ...

Abstract	Cervus albirostris
MeSH term(s)	Animals ; China ; Deer/genetics ; Genetic Variation/genetics ; Heterozygote ; Microsatellite Repeats/genetics
Language	English
Publishing date	2020-10-15
Publishing country	England
Document type	Journal Article
ZDB-ID	2043243-4
ISSN	1532-2378 ; 1049-5398
ISSN (online)	1532-2378
ISSN	1049-5398
DOI	10.1080/10495398.2020.1832104
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Article ; Online: lncRNA Sequencing of Antler Mesenchymal Tissue Revealed that the Regulatory Network of Antler Cell Proliferation and Differentiation.

Han, Ruobing / Han, Lei / Xia, Yanling / Guo, Mengya / Li, Heping

Animal biotechnology

2021 , Page(s) 1–10

Abstract	Antlers have been widely studied due to their unique physiological characteristics, such as rapid growth, periodic shedding and regeneration. However, little is known about how antler growth is regulated by long non-coding RNA (lncRNA). The aim of the present study was to identify the lncRNAs expression profile and explore the function of lncRNAs during the antler growth. Herein, RNA-sequencing technology (RNA-seq) was performed on the three growth periods (early developmental period: EP, middle developmental period: MP, later developmental period: LP) of male sika deer (
Language	English
Publishing date	2021-05-19
Publishing country	England
Document type	Journal Article
ZDB-ID	2043243-4
ISSN	1532-2378 ; 1049-5398
ISSN (online)	1532-2378
ISSN	1049-5398
DOI	10.1080/10495398.2021.1924762
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Haplotype-resolved Genome of Sika Deer Reveals Allele-specific Gene Expression and Chromosome Evolution.

Han, Ruobing / Han, Lei / Zhao, Xunwu / Wang, Qianghui / Xia, Yanling / Li, Heping

Genomics, proteomics & bioinformatics

2022 Volume 21, Issue 3, Page(s) 470–482

Abstract: Despite the scientific and medicinal importance of diploid sika deer (Cervus nippon), its genome resources are limited and haplotype-resolved chromosome-scale assembly is urgently needed. To explore mechanisms underlying the expression patterns of the ... ...

Abstract	Despite the scientific and medicinal importance of diploid sika deer (Cervus nippon), its genome resources are limited and haplotype-resolved chromosome-scale assembly is urgently needed. To explore mechanisms underlying the expression patterns of the allele-specific genes in antlers and the chromosome evolution in Cervidae, we report, for the first time, a high-quality haplotype-resolved chromosome-scale genome of sika deer by integrating multiple sequencing strategies, which was anchored to 32 homologous groups with a pair of sex chromosomes (XY). Several expanded genes (RET, PPP2R1A, PPP2R1B, YWHAB, YWHAZ, and RPS6) and positively selected genes (eIF4E, Wnt8A, Wnt9B, BMP4, and TP53) were identified, which could contribute to rapid antler growth without carcinogenesis. A comprehensive and systematic genome-wide analysis of allele expression patterns revealed that most alleles were functionally equivalent in regulating rapid antler growth and inhibiting oncogenesis. Comparative genomic analysis revealed that chromosome fission might occur during the divergence of sika deer and red deer (Cervus elaphus), and the olfactory sensation of sika deer might be more powerful than that of red deer. Obvious inversion regions containing olfactory receptor genes were also identified, which arose since the divergence. In conclusion, the high-quality allele-aware reference genome provides valuable resources for further illustration of the unique biological characteristics of antler, chromosome evolution, and multi-omics research of cervid animals.
MeSH term(s)	Animals ; Deer/genetics ; Alleles ; Haplotypes ; Chromosomes ; Gene Expression
Language	English
Publishing date	2022-11-15
Publishing country	England
Document type	Journal Article
ZDB-ID	2240213-5
ISSN	2210-3244 ; 1672-0229
ISSN (online)	2210-3244
ISSN	1672-0229
DOI	10.1016/j.gpb.2022.11.001
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Article: Quantitative patient graph analysis for transient ischemic attack risk factor distribution based on electronic medical records.

Wen, Jian / Zhang, Tianmei / Ye, Shangrong / Zhang, Peng / Han, Ruobing / Chen, Xiaowang / Huang, Ran / Chen, Anjun / Li, Qinghua

Heliyon

2023 Volume 10, Issue 1, Page(s) e22766

Abstract: A transient ischemic attack (TIA) affects millions of people worldwide. Although TIA risk factors have been identified individually, a systemic quantitative analysis of all health factors relevant to TIA using electronic medical records (EMR) remains ... ...

Abstract	A transient ischemic attack (TIA) affects millions of people worldwide. Although TIA risk factors have been identified individually, a systemic quantitative analysis of all health factors relevant to TIA using electronic medical records (EMR) remains lacking. This study employed a data-driven approach, leveraging hospital EMR data to create a TIA patient health factor graph. This graph consisted of 737 TIA and 737 control patient nodes, 740 health factor nodes, and over 33,000 relations between patients and factors. For all health factors in the graph, the connection delta ratios (CDRs) were determined and ranked, generating a quantitative distribution of TIA health factors. A literature review confirmed 56 risk factors in the distribution and unveiled a potential new risk factor "rhinosinusitis" for future validation. Moreover, the patient graph was visualized together with the TIA knowledge graph in the Unified Medical Language System. This integration enables clinicians to access and visualize patient data and international standard knowledge within a unified graph. In conclusion, graph CDR analysis can effectively quantify the distribution of TIA risk factors. The resulting TIA risk factor distribution might be instrumental in developing new risk prediction machine learning models for screening and early detection of TIA.
Language	English
Publishing date	2023-11-25
Publishing country	England
Document type	Journal Article
ZDB-ID	2835763-2
ISSN	2405-8440
ISSN	2405-8440
DOI	10.1016/j.heliyon.2023.e22766
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