1.Gene clone, eukaryotic expression vector and tissue expression profile analysis of porcine BST-2 gene
Ning KONG ; Yongguang WU ; Qiong MENG ; Zhongze WANG ; Wu TONG ; Hao ZHENG ; Guoxin LI ; Tongling SHAN ; Enmin ZHOU ; Guangzhi TONG
Chinese Journal of Veterinary Science 2017;37(8):1594-1599,1640
In order to study the biological function of pig BST-2 gene,the BST-2 gene was amplified with specific primers from porcine kidney tissue,and molecular characterization of BST-2 nuclectide and amino acid sequence were analyzed with bioinformatics tools and online server.Then the prokaryotic expression and tissue expression profile analysis was carried out.The results showed that the full length of pig BST-2 gene was 851 bp and contained 23 bp of 5'-UTR,294 bp of 3'-UTR and 534 bp of CDS and the gene encoded 177 aa.Amino acid sequence analysis of pig BST-2 protein showed 46.1% identity with gorilla gorilla,41.7% with cricetulus griseus,39.5% with mus musculus,35.4% with equus asinus,42.0% with felis catus,40.5% with bos mutus,44.4% with macaca mulatta,38.7% with ovis aries and 46.8% with homo sapiens.BST-2 protein contained 2 transmembrane structure (27-49 aa and 154-176 aa),2 glycosylation sites and 14 potential phosphorylation sites including ATM,CK Ⅱ,PKA,PKC binding sites.The pig BST-2 protein was expressed in Vero cells after translated the recombinant plasmid FLAG-BST-2.Semiquantitative PCR results showed that BST-2 gene was expressed in all the tissues,especially in lymph nodes,thymus,tonsils,spleen,large intestine and small intestine.This study provide a foundation for further understanding the antiviral mechanism of pig BST-2 protein.
2.Oxidative stress in liver tissues in HCC patients after TACE
Hao SU ; Guangzhi ZHU ; Hongqiang LIN ; Yi LIN ; Yizhen GONG ; Jiaquan LI ; Zhiming LIU ; Lequn LI ; Tangwei LIU ; Zili Lü ; Lünan YAN ; Tao PENG
Chinese Journal of General Surgery 2009;24(10):795-798
Objective To investigate the levels of oxidative stress in liver tissues of hepatocelluar carcinoma(HCC)patients after transcatheter arterial chemotherapy(TAC).Methods Immunohistochemistry streptavidin biotinylated peroxidase(S-P)method was used to detect the cellular levels of 8-hydroxy-2'-deoxyguanosine(8-OHdG),p53 and p21~(waf1/cip1).Eighty-nine HCC patients were divided into TAC group(39 cases)and Non-TAC group(50 cases).15 Non-HCC liver tissues served as controls.Result 8-OHdG level was higher in Non-TAC group than that in TAC group in tumor tissues (F=9.516,P<0.05),with that being the lowest in control group(F=9.516,P<0.01);8-OHdG levels in cancer tissues were significantly higher than that in tumor surrounding tissues in both TAC group (t=7.101,P<0.001)and Non-TAC group(t=8.020,P<0.001),there was no significant difference of 8-OHdG levels between para-tumor tissues and controls.The levels of 8-OHdG between tumor and its surrounding tissues in TAC group(r=0.651,P<0.001)and non-TAC group(r=0.493,P<0.01)was in positive correlation.The difference of p53 levels in cancer tissues in TAC group and Non-TAC group were not statistically significant and p53 was not detected in para-tumor tissues.The difference of p21~(waf1/cip1) levels among TAC group,Non-TAC group and controls was statistically significant,the levels of p21~(waf1/cip1) in normal group was the highest(F=13.459,P<0.001),followed by that in TAC and Non-TAC group in cancer tissues(TAC vs.Non-TAC group,P<0.01);p21~(waf1/cip1) expression in normal controls was significantly higher than that in both TAC and Non-TAC group in para-tumor tissues(F=16.613,P<0.001).The correlation of p21 ~(waf1/cip1) levels between tumor and its surrounding tissues was significant in non-TAC group(r=0.872,P<0.001).Conclusions Oxidative stress levels in HCC tumor tissues were higher than in para-tumor tissues and non-HCC liver tissues.Cancer cells probably survive chemotherapy by fortifying oxidative stress repair mechanism.
3.Meta analysis of hearing loss caused by the combined effect of noise and heat in the working population
Mengyao CHEN ; Hao CHEN ; Huimin WANG ; Guangzhi YANG ; Enmin DING ; Baoli ZHU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2022;40(6):419-422
Objective:To explore the effect of the combined effect of noise and heat on occupational hearing loss of workers by using Meta-analysis method.Methods:In August 2020, the Chinese and English literature on the relationship between exposure to noise and heat and occupational hearing loss published from January 2005 to August 2020 by CNKI, China Biomedical Literature Service System, Wanfang Data Knowledge Service Platform, VIP Official Database, Medline and PubMed Databases were searched, using noise, heat or hyperthermia, hearing as keywords. The selected data were analyzed by Stata 12.0 software, and the combined OR (95% CI) value included in the literature was calculated. Sensitivity analysis was used to explore the source of heterogeneity and analyze publication bias. Results:A total of 14 literatures (14 in Chinese, 0 in English) were included in the analysis, and 38654 subjects were included, including 6411 workers in the noise and heat combined effect group and 32243 workers in the noise alone group. The probability of hearing loss in the noise and heat combined effect group was 1.39 times higher than that in the noise alone group (95% CI: 1.14-1.69). The effect size OR was stable after sensitivity analysis, and there was no publication bias in the included literatures tested by Egger's and Begg's Method ( z=0.38, P=0.702, t=-0.74, P=0.476) . Conclusion:Simultaneous exposure to noise and heat may increase the risk of hearing loss for workers in noisy workplaces.
4.Transcriptomic analysis of delayed cerebral ischemia after aneurysmal subarachnoid hemorrhage
Guangzhi HAO ; Yuwei HAN ; Da HUO
Chinese Journal of Neuroanatomy 2024;40(3):341-346
Objective:To analyze the key differentially expressed genes and related pathways in patients with de-layed cerebral ischemia(DCI)occurring after aneurysmal subarachnoid hemorrhage(aSAH)based on gene expression database(GEO)datasets.Methods:Gene datasets meeting the study requirements were screened from the GEO data-base and divided into a patient group with DCI after aSAH and a control group without DCI,and differentially expressed genes(DEGs)analysis,gene ontology(GO)enrichment analysis,Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis,and gene set enrichment analysis(GSEA)were performed by using R software to find rele-vant genes and pathways.Results:A total 140 differentially expressed genes were identified.KEGG analysis showed that there were 10 significantly enriched signaling pathways(P<0.05),mainly related to hypoxia-inducible factor-1(HIF-1)signaling pathway,glutamatergic synapses,cell adhesion molecules,and chemokine signaling pathways,etc.Twenty-six significantly enriched entries were obtained according to the functional analysis of GO,which involved entries in three categories,namely,biological processes,cellular components,and molecular functions.GSEA analysis showed that two pathways,extracellular matrix receptor interactions as well as peroxisomes,had significant enrichment differ-ences between the patient group and the control group.Conclusion:The development of DCI after aSAH involves sever-al genes such as COL17A1,RPL11,FCAMR,GNB2L1,HIF1A,and can affect the development of DCI through various pathways such as vascular dysfunction,inflammatory response,neurological injury,and oxidative stress.
5.Intelligent precision assistant technology in spinal cord stimulation after spinal cord injury: application and research progress
Mi ZHOU ; Hao ZHONG ; Song LIU ; Cong XING ; Guangzhi NING
Chinese Journal of Trauma 2023;39(9):857-864
Spinal cord injury is a severe central nervous system disorder that often results in sensory and motor impairments below the level of injury, accompanied by respiratory failure, gastrointestinal disturbances, and hemodynamic instability. Existing treatment such as early decompression surgery and steroid-based shock therapy has limited effectiveness and can lead to serious complications. Meanwhile, the rehabilitation therapy is costly and can only provide limited functional recovery. Spinal cord stimulation (SCS), by stimulating and regulating residual neurons in the lower spinal cord, can effectively promote motor function recovery in patients with spinal cord injury. The introduction of intelligent precision-assistive technologies can help to create more precise and individualized neuro-regulation and rehabilitation strategies tailored to the severity, segment, and course of spinal cord injury. These strategies, coupled with real-time feedback and continuous algorithm adjustments, enabled remote and secure control of SCS, enhancing the treatment effectiveness and safety for patients with spinal cord injury, which can help to develop more effective rehabilitation intervention plans and further increase the clinical translational value. In this review, the authors summarized the research progress in the application of intelligent precision-assistive technology in SCS for the treatment of spinal cord injury, so as to provide valuable insights for the rehabilitation of SCI.
6.Meta analysis of hearing loss caused by the combined effect of noise and heat in the working population
Mengyao CHEN ; Hao CHEN ; Huimin WANG ; Guangzhi YANG ; Enmin DING ; Baoli ZHU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2022;40(6):419-422
Objective:To explore the effect of the combined effect of noise and heat on occupational hearing loss of workers by using Meta-analysis method.Methods:In August 2020, the Chinese and English literature on the relationship between exposure to noise and heat and occupational hearing loss published from January 2005 to August 2020 by CNKI, China Biomedical Literature Service System, Wanfang Data Knowledge Service Platform, VIP Official Database, Medline and PubMed Databases were searched, using noise, heat or hyperthermia, hearing as keywords. The selected data were analyzed by Stata 12.0 software, and the combined OR (95% CI) value included in the literature was calculated. Sensitivity analysis was used to explore the source of heterogeneity and analyze publication bias. Results:A total of 14 literatures (14 in Chinese, 0 in English) were included in the analysis, and 38654 subjects were included, including 6411 workers in the noise and heat combined effect group and 32243 workers in the noise alone group. The probability of hearing loss in the noise and heat combined effect group was 1.39 times higher than that in the noise alone group (95% CI: 1.14-1.69). The effect size OR was stable after sensitivity analysis, and there was no publication bias in the included literatures tested by Egger's and Begg's Method ( z=0.38, P=0.702, t=-0.74, P=0.476) . Conclusion:Simultaneous exposure to noise and heat may increase the risk of hearing loss for workers in noisy workplaces.
7.Molecular characterization of Japanese encephalitis virus strains prevalent in Chinese swine herds.
Hao ZHENG ; Tongling SHAN ; Yu DENG ; Chunqing SUN ; Shishan YUAN ; Yang YIN ; Guangzhi TONG
Journal of Veterinary Science 2013;14(1):27-36
Japanese encephalitis virus (JEV) is the leading cause of viral encephalitis in Asia and domestic pigs serve as the amplifying hosts. In the present study, the full genomic sequences of two JEV strains (HEN0701 and SH0601) isolated from pigs in China were determined and compared with other 12 JEV strains deposited in GenBank. These two strains had an 88.8% nucleotide sequence similarity and 97.9% deduced amino acid sequence homology. HEN0701 had high nucleotide sequence and high amino acid sequence identity with genotype I (GI) strains, while SH0601 had high nucleotide sequence and high amino acid sequence identity with GIII strains at both the gene and full genome levels. Further phylogenetic analysis showed that HEN0701 belonged to the JEV GI group and SH0601 was classified as a GIII strain. Analysis of codon usage showed there were a few differences between the GI and GIII strains in nucleotide composition and codon usage for the open reading frames.
Animals
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Cell Line
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Cricetinae
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Encephalitis Virus, Japanese/classification/*genetics
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Encephalitis, Japanese/epidemiology/*veterinary/virology
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Gene Expression Regulation, Viral/physiology
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Genome, Viral
;
Molecular Epidemiology
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Phylogeny
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Swine
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Swine Diseases/epidemiology/*virology
8.Molecular characterization of Japanese encephalitis virus strains prevalent in Chinese swine herds.
Hao ZHENG ; Tongling SHAN ; Yu DENG ; Chunqing SUN ; Shishan YUAN ; Yang YIN ; Guangzhi TONG
Journal of Veterinary Science 2013;14(1):27-36
Japanese encephalitis virus (JEV) is the leading cause of viral encephalitis in Asia and domestic pigs serve as the amplifying hosts. In the present study, the full genomic sequences of two JEV strains (HEN0701 and SH0601) isolated from pigs in China were determined and compared with other 12 JEV strains deposited in GenBank. These two strains had an 88.8% nucleotide sequence similarity and 97.9% deduced amino acid sequence homology. HEN0701 had high nucleotide sequence and high amino acid sequence identity with genotype I (GI) strains, while SH0601 had high nucleotide sequence and high amino acid sequence identity with GIII strains at both the gene and full genome levels. Further phylogenetic analysis showed that HEN0701 belonged to the JEV GI group and SH0601 was classified as a GIII strain. Analysis of codon usage showed there were a few differences between the GI and GIII strains in nucleotide composition and codon usage for the open reading frames.
Animals
;
Cell Line
;
Cricetinae
;
Encephalitis Virus, Japanese/classification/*genetics
;
Encephalitis, Japanese/epidemiology/*veterinary/virology
;
Gene Expression Regulation, Viral/physiology
;
Genome, Viral
;
Molecular Epidemiology
;
Phylogeny
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Swine
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Swine Diseases/epidemiology/*virology
9.Effect of melatonin on reducing mitochondrial oxidative stress in mice with ischemia-reperfusion via a silent information regulator
Yangyang LIU ; Guobiao LIANG ; Endi ZHEN ; Chongdan MAO ; Xu GAO ; Guangzhi HAO ; Yushu DONG
Chinese Journal of Cerebrovascular Diseases 2017;14(10):519-524
Objective To investigate the effect of melatonin on mice with ischemia-reperfusion via a silent information regulator 1 (SIRT1) reducing mitochondrial oxidative stress mechanism. Methods A transient middle cerebral artery occlusion ( MCAO) cerebral ischemia-reperfusion ( IR) model in mice was established by the suture-occluded method. One hundred and ninety mice were injected with melatonin intraperitoneally or the SIRT1 inhibitor (EX527) intracerebroventricularly,30 dead and model failure mice were excluded. They were divided into IR,melatonin,melatonin +EX527,and EX527 groups (n =40 in each group ) according to the random number table. The cerebral infarct volume was detected by the triphenyltetrazolium chloride (TTC) method,the brain edema was measured by the wet and dry weight method and the neurological deficit scores were measured. Western blot was used to detect SIRT1,Ac-P53, acetylated-nuclear factorκB (Ac-NF-κB),BCl2,Bax proteins in the mitochondria and cytoplasm,as well as the cytochrome C protein expression. A single factor analysis of variance was used for comparison among the groups. Results ( 1 ) There were significant differences in cerebral infarction volume, neurological dysfunction scores and cerebral edema among the four groups ( F values,16. 452,23. 622,and 18. 786, respectively (all P<0. 05). There were significant differences in the expression levels of SIRT1,Ac-P53, Ac-NF-κB,BCl2, and Bax among the four groups ( F values, 2348. 158, 1434. 841, 7042. 563, 14627. 128,and 691. 475,respectively,all P<0. 05). There were significant differences in mitochondrial membrane potential,mitochondrial reactive oxygen species,and complex I activity in mice among the four groups (F value,28. 454,33. 728 and 29. 716,respectively,all P <0. 05). (2) Compared with the IR group,the infarct volume was reduced (32 ± 5 mm3 vs. 57 ± 5 mm3,P<0. 05),neurological deficit scores were decreased (2. 4 ± 0. 3 vs. 3. 5 ± 0. 3,P<0. 05);brain edema was reduced (80. 2 ± 0. 9% vs. 83. 9 ± 1. 2%,P<0. 05);the expression levels of SIRT1 and anti-apoptosis protein BCL2 were increased in the melatonin group (P<0. 05);the expression levels of pro-apoptotic protein BAX and Ac-P53,Ac-NF-κB were reduced ( P <0. 05 );the mitochondrial membrane potential, mitochondrial complex I activity and cytochrome C level were increased (P<0. 05);and the cytoplasmic reactive oxygen species and cytochrome C level were decreased (P < 0. 05). (3) Compared with the melatonin group,cerebral infarction volume were increased (42 ± 5 mm3 vs. 32 ± 5 mm3,P < 0. 05);nerve dysfunction scores were increased(3. 2 ± 0. 3 vs. 2. 4 ± 0. 3,P<0. 05);cerebral edema was aggravated (83. 4 ± 0. 8% vs. 80. 2 ± 0. 9%, P < 0. 05 );the expression levels of SIRT1 and anti-apoptotic protein BCL2 were reduced (P <0. 05);the pro-apoptotic protein BAX,Ac-P53,and Ac-NF-κB expression levels were increased (P<0. 05);the mitochondrial membrane potential and mitochondrial complex I activity and cytochrome C level were decreased (P<0. 05);and the cytoplasmic reactive oxygen species and cytoplasmic cytochrome C level were increased in the melatonin+EX527 group (P<0. 05). Conclusion In ischemic stroke model mice, melatonin plays a neuroprotective role by activating the SIRT1 signaling pathway and reducing oxidative stress injury and cell death in mitochondria,thus plays a role in cerebral protection.
10.The effects of melatonin therapy on hypothalamic-pituitary-adrenal axis in rats with posttraumatic stress disorder
Xiao SUN ; Zhida LI ; Guangzhi HAO ; Yushu DONG
Chinese Journal of Neuroanatomy 2023;39(6):677-681
Objective:To investigate the effects of melatonin(MLT)on hypothalamic-pituitary-adrenal(HPA)axis in posttraumatic stress disorder(PTSD)rats induced by foot shocks.Methods:The rat model of PTSD was prepared by plantar electroshock,and MLT was given by intraperitoneal injection.The behavioral changes of rats were detected by rejection reaction test,the mRNA expression of corticotropin-releasing hormone(CRH)in hypothalamus was detected by real time RT-PCR,and the contents of adrenocorticotropin hormone(ACTH),epinephrine(EPI)and glucocorticoid(GC)in serum were detected by enzyme-linked immunosorbent assay(ELISA).Results:In the PTSD group,the rejection reaction was obvious,the expression of CRH mRNA in hypothalamus was increased(P<0.05),the serum ACTH and EPI were increased(P<0.05),but the GC level was decreased(P<0.05).After MLT treatment,the rejection reaction of PTSD rats was significantly alleviated,CRH mRNA expression in hypothalamus was decreased(P<0.05),serum ACTH and EPI levels were decreased(P<0.05),and GC levels were increased(P<0.05).Conclusion:MLT treatment can relieve the symptoms of PTSD and restore the neuroendocrine balance of HPA axis in rats.