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.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.
4.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
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Molecular Epidemiology
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Phylogeny
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Swine
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Swine Diseases/epidemiology/*virology
5.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
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Molecular Epidemiology
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Phylogeny
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Swine
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Swine Diseases/epidemiology/*virology
6.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.
7.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.
8.Establishment controlled motor cortex impact injury model of rats and detection of motor dysfunction
Weilong SONG ; Guangzhi HAO ; Zhen WANG ; Hai JIN
Chinese Journal of Neuroanatomy 2023;39(6):705-708
Objective:This study was to establish a rat model of controlled motor cortex impact injury(CCI).Methods:SD rat CCI model was prepared by electromagnetic shock method.A neurologic severity score(NSS)was employed to evaluate the status of the rat after injury.The changes of neuron-specific enolase(NSE)in serum were detected by ELISA.The sensomotor function was detected by sticker removal test,and the fine motor function was de-tected by food pellets grasping test.Results:The serum NSE level of rats increased 1 h after CCI trauma,and reached the highest level 6 h after CCI trauma.The sensomotor function and fine motor function of CCI model rats were signifi-cantly impaired.Conclusion:The CCI model of rat motor cortex was successfully established,and the rats showed obvious motor dysfunction.
9.DDX3X/NF-κB pathway mediates early neurons apoptosis in mice with subarachnoid hemorrhage
Guangzhi HAO ; Yu HUAN ; Yuwei HAN ; Yushu DONG ; Guobiao LIANG
Chinese Journal of Neuroanatomy 2024;40(1):85-90
Objective:To study the role of DDX3X/NF-κB pathway in early neuronal apoptosis in subarachnoid hemorrhage(SAH)mice.Methods:The mouse model of SAH was established by internal carotid artery puncture,and the neurological function score of the mice was evaluated.The DDX3X expression was knocked down using recombinant lentivirus expressing DDX3X targeted shRNA(Lv-shDDX3X),or the NF-κB pathway was inhibited by NF-κB-IN-1(IN-1).Western Blot was used to detect the expression of DDX3X and NF-κB(p65)in mouse cortex.TUNEL/NeuN staining was used to detect the apoptosis of cerebral cortex neurons.Results:Twenty-four hours after SAH operation,the neurological function of mice was significantly impaired(P<0.05).While the expression of DDX3X was signifi-cantly increased and the expression of NF-κB(p65)was significantly decreased in the cortex(P<0.05).When the DDX3X expression is knocked down firstly,then SAH surgery is performed.The neurological function of mice was sig-nificantly recovered,and the expression of NF-κB(p65)protein was significantly higher than that in SAH group(P<0.05);If the NF-κB activity was inhibited by IN-1 while DDX3X knockdown,there is no significant recovery of neuro-logical function in SAH mice.TUNEL/NeuN staining showed that the number of TUNEL-positive neurons in the brain tissue after DDX3X knockdown was less than that in the SAH group(P<0.05),while the number of TUNEL-positive neurons was not significantly reduced when IN-1 was used to inhibit NF-κB activity at the same time of DDX3X knock-down.Conclusion:DDX3X/NF-κB mediated cell death in mice with early brain injury after SAH.
10.Clinical application of non-invasive myocardial work imaging in ST-segment elevation myocardial infarction patients with left ventricular remodeling after percutaneous coronary intervention
Xue YANG ; Quanrui MA ; Yanping XU ; Ying WANG ; Wei CAO ; Guangzhi CONG ; Yu HAO ; Lisha NA
Chinese Journal of Ultrasonography 2023;32(6):493-500
Objective:To explore the application value of non-invasive myocardial work imaging in evaluating the cardiac function of ST-segment elevation myocardial infarction (STEMI) patients with left ventricular remodeling (LVR) after percutaneous coronary intervention (PCI).Methods:One hundred and twenty-six patients with STEMI undergoing PCI in General Hospital of Ningxia Medical University from December 2021 to September 2022 were prospectively collected and divided into left ventricular remodeling group (LVR group, 34 cases) and non left ventricular remodeling group (NLVR group, 92 cases) according to whether there was left ventricular remodeling 3 months after surgery. General data were collected. Routine echocardiography and noninvasive myocardial work imaging were performed before, 1 week, 1 month, and 3 months after surgery, the differences in the above parameters between the two groups were compared. Pearson correlation analysis was used to analyze the correlation between the indicators.Logistic regression analysis was used to determine the independent risk factors of left ventricular remodeling after STEMI, and a predictive model was obtained. The diagnostic value of the model was judged by ROC curve.Results:①General information comparison: There were statistically significant differences between the two groups in BMI, systolic blood pressure, diastolic blood pressure, average number of stents implanted, and history of hyperlipidemia (all P<0.05), but there was no significant difference in other data (all P>0.05). There was no statistically significant difference in two-dimensional transthoracic echocardiography (2D-TTE) parameters and non-invasive myocardial work (MW) parameters between the two groups before and 1 week after operation (both P>0.05). ②2D-TTE parameter comparison: LVESV and LVEDV at 3 months after PCI in the LVR group were significantly higher than those in the NLVR group, and LVEF and E/A were significantly lower than those in the NLVR group (all P<0.05); There were no significant differences in other indexes between the two groups by conventional echocardiography at 3 months after PCI(all P>0.05). ③Comparisons of noninvasive myocardial work parameters: GLS, GWE, GWI, GCW at 1 month and 3 months after PCI in the LVR group were significantly lower than those in the NLVR group, and GWW were significantly higher than those in the NLVR group ( P<0.001). ④Correlation analysis: GLS, GWE, GCW, GWI and LVEDV were negatively correlated at 1 month after operation ( r=-0.42, -0.38, -0.50, -0.53, all P<0.001), GWW was positively correlated with LVEDV ( r=0.45, P<0.001). ⑤Logistic regression analysis: GLS<17%, GCW<1 900 mmHg%, GWW>105 mmHg%, and GWE<90 mmHg% at 1 month after PCI were independent predictors for LVR in STEMI patients after PCI (all P<0.05). The predictive model was Logit (P)=0.692GLS+ 0.804GCW+ 0.972GWW+ 0.880GWE. The AUC of this model was 0.886, 95% CI=0.845-0.926, which was significantly higher than single index, the sensitivity was 0.86, and the specificity was 0.79. Conclusions:GLS, GWE, GWI, GCW are positively correlated with LVR, while GWW is negatively correlated with left ventricular remodeling. Noninvasive myocardial work parameters are independent risk factors for left ventricular remodeling in patients with STEMI after PCI surgery. This technique can be used to evaluate LVR and has great clinical application value.