1.Analysis of Potential Suitable Areas and Key Ecological Factors of Polygonatum odoratum Based on MaxEnt Model
Anling HUANG ; Jinxiang JIANG ; Zhiqin REN ; Youqiong HU ; Zhiwei WANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(18):178-185
		                        		
		                        			
		                        			ObjectiveThe potential suitable area for ecological planting, key ecological factors, and suitable range of Polygonatum odoratum in China were analyzed to provide theoretical and scientific guidance for the artificial planting of P. odoratum. MethodA total of 454 geographical distribution records of P. odoratum in China and 118 ecological factors were used in this study. The maximum entropy model (MaxEnt) was adopted to predict the suitable areas of P. odoratum. The key ecological factors and their suitable ranges were analyzed by the jackknife method, contribution rates of ecological factors, and response curves. ResultThe suitable areas of P. odoratum were mainly located in the northwest, north, and northeast of China, the highly suitable areas of which were concentrated in Shaanxi, Shanxi, Gansu, etc. Solar radiation in November (Srad11), precipitation in July (Prec7), percentage of evergreen/deciduous needleleaf trees (Class1), silt content (2-50 μm) mass fraction (SLTPPT), and annual average temperature (Bio1) were found to be the key ecological factors affecting the suitable distribution of P. odoratum in China. The cumulative contribution rate of solar radiation factors (31.29%)>vegetation factors (25.61%)>soil factors (19.52%)>precipitation factors (11.38%)>temperature factors (8.57%)>topography factors (3.63%). ConclusionIt is suggested to carry out ecological planting of P. odoratum mainly in Shaanxi (such as Baoji and Ankang Cities and Ningshan, Liuba, and Hua Counties), Gansu (such as Tianshui City, Gannan Tibetan Autonomous Prefecture, and Liangdang and Huating Counties), and Shanxi (such as Yangquan, Taiyuan, Fenyang, and Jinzhong Cities, as well as Xingxian County) of China. Solar radiation factors should be given priority in the planting process, followed by vegetation, soil, precipitation, temperature, and topography factors. The range of key ecological factors, namely Srad11, Prec7, Class1, SLTPPT, and Bio1 should be controlled within 8 095.21-10 334.98 (optimum 8 787.50) kJ·m-2·d-1, 109.99-223.60 (146.91) mm, 1.00%-9.45% (6.76%-10.68%), 41.73%-50.35% (46.53%), and 3.29-16.33 (13.57) °C, respectively. 
		                        		
		                        		
		                        		
		                        	
2.Immune effect of H9N2 subtype AIV NP protein by prokaryotic expression
Xiaofeng LI ; Zhixun XIE ; Zhihua RUAN ; Meng LI ; Dan LI ; Minxiu ZHANG ; Zhiqin XIE ; Sisi LUO ; You WEI ; Liji XIE ; Tingting ZENG ; Yanfang ZHANG ; Jiaoling HUANG ; Sheng WANG
Chinese Journal of Veterinary Science 2024;44(6):1113-1119
		                        		
		                        			
		                        			The aim of this study is to investigate the immune effect of H9 subtype avian influenza virus(AIV)NP protein on mice and lay the foundation for the development of avian influenza vi-rus(AIV)vaccine.The H9N2 virus NP gene amplification product was cloned into the pET-32a expression vector,and the protein expression was verified by SDS-PAGE and Western blot,and the immune effect was evaluated by measuring the secretion of supernatant multicytokines in mouse splenocytes culture.The results showed that the total length of the coding region sequence of NP gene was 1 497 bp,NP recombinant proteins exist in both soluble and insoluble protein forms,and the specific bands were visible in Western blot.After immunizing mice,serum produces IgG-bind-ing antibodies with antibody titers of 1∶40 000.Compared with the control group,IL-2,IL-5 and IL-13 were significantly increased(P<0.001),and the secretion of IL-6 was significantly increased compared with the control group.IL-4 and IL-12 p70 secretions were elevated compared with con-trols,but there was no significant difference.Compared with the control group,the secretions of IL-1β,IL-18,GM-CMF,TNF-α and IFN-γ were inhibited,but the difference was not significant(P>0.05).The results showed that NP recombinant protein is a good immunogen,laying a foundation for in-depth research on influenza vaccine.
		                        		
		                        		
		                        		
		                        	
3.Preparation of monoclonal antibody against σA protein of avian reovirus and es-tablishment of sandwich ELISA method for detection
Bingyi YANG ; Zhixun XIE ; Zhiqin XIE ; Hongyu REN ; You WEI ; Liji XIE ; Jiaoling HUANG ; Sheng WANG
Chinese Journal of Veterinary Science 2024;44(7):1373-1379
		                        		
		                        			
		                        			In order to prepare monoclonal antibody to σ A protein of avian reovirus(ARV)and es-tablish a sandwich ELISA method for the detection of ARV pathogens.In this study,the σ A pro-tein of ARV was expressed as antigen by prokaryotic expression and used to immunize BALB/c mice.Then,stable hybridoma cell lines were screened,and monoclonal antibodies were prepared.A sandwich ELISA detection method based on monoclonal antibody of σA protein was established,and the sensitivity,specificity,repeatability,and accuracy were tested.The results showed that the recombinant plasmid pET-32a-σA was successfully constructed and well expressed in Escherichia coli.After immunizing mice,two hybridoma cell lines 6B3 and 8E11,which could secrete mono-clonal antibodies stably,were successfully prepared.Both monoclonal antibodies could react with natural ARV.One of the monoclonal antibodies secreted by 6B3 was selected as the capture anti-body and the ARV-positive chicken polyclonal antibody was used as the detection antibody.A sand-wich ELISA method was established to detect ARV by optimizing the reaction conditions.The specific test showed that the method only detected ARV pathogens and no other common chicken viral pathogens were detected.The detection limit was 7.72 X 102 EID50/mL of ARV antigen.The coefficient of variation of the intra-and inter-assay tests were less than 5.0%and the reproducibili-ty was good.Thirty samples were tested simultaneously by σA-sandwich ELISA and PCR,and the results were consistent with each other.In conclusion,a sandwich ELISA method based on the monoclonal antibody of σA protein was successfully established for the identification and detection of ARV,which provided a technical means for the accurate and rapid detection of ARV.
		                        		
		                        		
		                        		
		                        	
4.Research progress of molecular diagnosis and treatment strategies for RCBTB1 gene-related inherited retinal disease
Chinese Journal of Ocular Fundus Diseases 2024;40(6):472-477
		                        		
		                        			
		                        			RCBTB1 gene associated hereditary retinopathy is an extremely rare inherited retinal disease (IRD) discovered recently. The mutation of RCBTB1 gene can lead to a variety of IRD clinical phenotypes, such as early retinitis pigmentosa and delayed chorioretinal atrophy. The hereditary mode of RCBTB1 gene associated retinopathy is autosomal recessive. RCBTB1 gene plays an important role in maintaining mitochondrial function and anti-oxidative stress defense mechanism of retinal pigment epithelium cells. In the future, it is necessary to further determine whether there is a genotypic and phenotypic correlation in the age of onset of RCBTB1 gene associated retinopathy or multi-organ involvement, and evaluate the safety and efficacy of adeno-associated virus-mediated RCBTB1 gene replacement therapy in animal models, to explore the feasibility of gene replacement therapy and stem cell therapy.
		                        		
		                        		
		                        		
		                        	
5.Construction of a "4M" death education intervention program for advanced cancer caregivers based on the theory of knowledge, belief and practice
Tian HUANG ; Danting ZHANG ; Jie JIN ; Zhiqin YIN
Chinese Journal of Practical Nursing 2023;39(20):1562-1569
		                        		
		                        			
		                        			Objective:To construct a death education intervention program for advanced cancer caregivers to improve the reference for death education for advanced cancer caregivers.Methods:Content analysis, semi-structured interview, Delphi expert consultation method were used to develop a preliminary death education program based on the theory of knowledge, belief, and behavior. From April to May 2022, fifteen experts from palliative care, life and death education, oncology nursing, psychological nursing and other related fields were selected for two rounds of expert consultation, and the contents of the program were revised and improved through preliminary experiments.Results:After two rounds of expert consultation, the results showed that the expert opinions tend to be unanimous. The authoritative coefficient of experts was 0.87, and the Kendall coordination coefficients of feasibility, validity and scientificity of the two rounds of consultation were 0.181, 0.303, 0.363 and 0.249, 0.355, 0.366, respectively (both P<0.05). The preliminary experiments revised and improved the intervention frequency and content, and finally formed a death education intervention program for advanced cancer caregivers which included four-stage progressive death themes: made an appointment with death, made a discussion on death, made an embrace with death and made friends with death. Conclusions:The process of constructing a death education program for advanced cancer caregivers is scientific, and the content is feasible, valid, and scientific. In addition, it is of great significance to promote death education in palliative care.
		                        		
		                        		
		                        		
		                        	
6.Breaking blood expelling stasis method accelerates hematoma resolution after hyperacute intracerebral hemorrhage and its potential mechanism
Bei LI ; Yuexiang ZHOU ; Zhiqin HUANG ; Lizhou LIN ; Zi LIANG
Journal of Chinese Physician 2022;24(1):53-58
		                        		
		                        			
		                        			Objective:To explore the efficacy of breaking blood expelling stasis method accelerates hematoma resolution after intracerebral hemorrhage (ICH) and its potential mechanism.Methods:63 ICH patients confirmed by computer tomography (CT) scan from August 2019 to February 2020 were selected as the research objects and randomly divided into control group ( n=29, routine treatment plus placebo) and observation group ( n=34, routine treatment plus breaking blood expelling stasis granules). The changes of neurological function and hematoma volume were observed before and after treatment. At the same time, the ICH rat model was constructed to observe the changes of neurobehavior and hematoma volume after the intervention of breaking blood expelling stasis granules. The expressions of peroxidase proliferator-activated-receptor γ(PPARγ), CD11b and CD36 in the surrounding tissues of hematoma were detected by Western blot on the third day after the intervention. Results:After two weeks of treatment, the National Institutes of Health Stroke Scale (NIHSS) score and hematoma volume of the two groups decreased (all P<0.05), and the NIHSS score and hematoma volume of the observation group were significantly lower than those of the control group (all P<0.05). In addition, the changes of NIHSS score and hematoma volume in the observation group before and after treatment were significantly greater than those in the control group ( P<0.01). In animal experiments, the hematoma volume in the breaking blood expelling stasis group on the 14th day after ICH was significantly smaller than that in the ICH group [(9.8±4.9)mm 3 vs (17.6±6.4)mm 3,P<0.05], and the reduction of hematoma in the breaking blood expelling stasis group on the 7th and 14th day was significantly larger than that in the ICH group [(4.6±2.9)mm 3 vs (-2.1±1.6)mm 3, (14.3±3.8)mm 3 vs (4.2±2.8)mm 3, all P<0.01]. The percentage of right turn on 3rd, 7th and 14th day and the modified Neurological Severity Score (mNSS) on 7th and 14th day in the breaking blood expelling stasis group were lower than those in the ICH group (all P<0.05). Western blot analysis showed that the expressions of CD11b, CD36 and PPARγ in the breaking blood expelling stasis group on the third day after ICH were significantly higher than those in the ICH group (CD11b: 0.78±0.12 vs 0.49±0.11, P<0.05; CD36: 1.16±0.16 vs 0.80±0.11, P<0.05; PPARγ: 0.78±0.11 vs 0.37±0.10, P<0.01). Conclusions:Breaking blood expelling stasis can effectively accelerate intracerebral hematoma clearance and improve neurological outcome after ICH, and the mechanism maybe probably mediated by activating PPARγ and enhanced CD36, CD11b upregulation on microglia/macrophages, which resulting in facilitating erythrocyte endogenous phagocytosis.
		                        		
		                        		
		                        		
		                        	
7.Molecular characterization of chicken anemia virus in Guangxi Province, southern China, from 2018 to 2020
Minxiu ZHANG ; Xianwen DENG ; Zhixun XIE ; Yanfang ZHANG ; Zhiqin XIE ; Liji XIE ; Sisi LUO ; Qing FAN ; Tingting ZENG ; Jiaoling HUANG ; Sheng WANG
Journal of Veterinary Science 2022;23(5):e63-
		                        		
		                        			 Background:
		                        			Chicken anemia virus (CAV) causes chicken infectious anemia, which results in immunosuppression; the virus has spread widely in chicken flocks in China. 
		                        		
		                        			Objectives:
		                        			The aim of this study was to understand recent CAV genetic evolution in chicken flocks in Guangxi Province, southern China. 
		                        		
		                        			Methods:
		                        			In total, 350 liver samples were collected from eight commercial broiler chicken farms in Guangxi Province in southern China from 2018 to 2020. CAV was detected by conventional PCR, and twenty CAV complete genomes were amplified and used for the phylogenetic analysis and recombination analysis. 
		                        		
		                        			Results:
		                        			The overall CAV-positive rate was 17.1%. The genetic analysis revealed that 84 CAVs were distributed in groups A, B, C (subgroups C1-C3) and D. In total, 30 of 47 Chinese CAV sequences from 2005-2020 belong to subgroup C3, including 15 CAVs from this study. There were some specific mutation sites among the intergenotypes in the VP1 protein. The amino acids at position 394Q in the VP1 protein of 20 CAV strains were consistent with the characteristics of a highly pathogenic strain. GX1904B was a putative recombinant. 
		                        		
		                        			Conclusions
		                        			Subgroup C3 was the dominant genotype in Guangxi Province from 2018–2020.The 20 CAV strains in this study might be virulent according to the amino acid residue analysis. These data help improve our understanding of the epidemiological trends of CAV in southern China. 
		                        		
		                        		
		                        		
		                        	
8.Extensive expansion of the chemical diversity of fusidane-type antibiotics using a stochastic combinational strategy.
Xiaojun SONG ; Jianming LV ; Zhiqin CAO ; Huiyun HUANG ; Guodong CHEN ; Takayoshi AWAKAWA ; Dan HU ; Hao GAO ; Ikuro ABE ; Xinsheng YAO
Acta Pharmaceutica Sinica B 2021;11(6):1676-1685
		                        		
		                        			
		                        			Fusidane-type antibiotics, represented by helvolic acid, fusidic acid and cephalosporin P
		                        		
		                        		
		                        		
		                        	
9.Research advances in cytokines associated with primary biliary cholangitis
Journal of Clinical Hepatology 2020;36(2):446-450
		                        		
		                        			
		                        			 Primary biliary cholangitis (PBC) is an autoimmune liver disease characterized by nonsuppurative inflammation of the small- and medium-sized bile ducts in the liver. The pathogenesis of PBC remains unclear, and immunoregulation may play a critical role. As the important components of the immune system, cytokines may be involved in the pathogenesis of PBC. This article reviews related research advances in recent years, including the roles of pro-inflammatory cytokines (interleukin-12, interleukin-17, interleukin-9, interleukin-8, tumor necrosis factor-α, and interleukin-6), the anti-inflammatory cytokine interleukin-10, and their signaling pathways in PBC, so as to deepen the understanding of the pathogenesis of PBC and explore new treatment methods. 
		                        		
		                        		
		                        		
		                        	
10.Expression of CLEC4G in liver disease and its correlation with hepatocellular carcinoma
Manling TANG ; Xiang CHEN ; Zhiqin XIE ; Meiyuan HUANG ; Hui LIN ; Zuiming JIANG
International Journal of Surgery 2020;47(3):164-168,封三
		                        		
		                        			
		                        			Objective:To investigate the expression level of C-type lectin domain family 4 member G ( CLEC4 G) in liver disease tissues and its correlation with the clinicopathological characteristics of hepatocellular carcinoma (HCC) patients. Methods:The cancer tissue and the corresponding adjacent tissues (at least 2 cm from the edge of the cancer tissue), cut in surgeries from January to December in 2019, of 40 HCC patients in Zhuzhou Central Hospital, as well as 10 normal liver tissue samples (seen as far away as possible from the edge of the cancer tissue with naked eyes) and 10 liver cirrhosis samples were analyzed retrospectively. The tumor genome atlas (TCGA) database was used to screen the HCC transcriptome data sets, and bioinformatics methods were used to make expression heat maps and box maps which can help analyze the difference of CLEC4 G in cancer and adjacent tissues. The mRNA expression level of CLEC4 G was detected by conducting real-time fluorescence quantitative PCR (qRT-PCR), and the protein expression level of CLEC4G was detected by immunohistochemistry (IHC). The measurement data were expressed as mean±standard deviation ( Mean± SD). Group t test was used for inter-group comparison. The counting information was expressed as a percentage (%). The χ2 test was adopted to analyze the correlation between CLEC4 G expression level and the clinicopathological features of patients. Results:The expression level of CLEC4 G in cancer tissues was significantly decreased in heat map compared with that in adjacent tissues. In the box figure, the relative expression of CLEC4 G mRNA in the cancer tissues was (82.5±18.9) and (3 354.4±296.2) in paracancer tissues, with statistically significant difference ( P<0.001). Respectively, qRT-PCR and IHC showed that mRNA of CLEC4 G were abundant in normal liver tissues (3 301.3±286.4), while they were very little in liver cancer tissues (63.6±32.9), significantly decreasing in liver cirrhosis (1 742.6±208.7) and paracancer tissues (1 553.2±249.9), with statistically significant difference ( P<0.001). Moreover, low CLEC4 G expression level was associated with tumor vascular metastasis in HCC patients. Conclusions:CLEC4 G is highly expressed in normal liver tissue, but with the progression of malignant liver disease, it is significantly decreased with little expression in HCC tissue. It can be expected to be a good marker for the pathological diagnosis of HCC.
		                        		
		                        		
		                        		
		                        	
            
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