1.Evaluation progress of the application of staplers in thoracoscopic lung surgery
Shenghui LI ; Yijiu REN ; Hang SU ; Minglei YANG ; Guofang ZHAO ; Yongxiang SONG ; Xuefei HU ; Deping ZHAO ; Qi XUE ; Chang CHEN
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2025;32(05):709-713
Compared to traditional suturing, lung stapling using automatic staplers offers advantages such as smaller trauma, faster wound healing, ease of operation, and lower complication rates, making it widely used in clinical practice. However, there are significant differences in bronchial tissue thickness at different anatomical locations, and the market is flooded with various types of staplers. Currently, there is a lack of recommended stapling schemes for bronchial staplers at different anatomical locations. This article reviews the development and application of automatic staplers and summarizes some types of staplers that are currently used in clinical practice, with the aim of promoting the formation of individualized stapler selection protocols for minimally invasive thoracic surgery based on the Chinese population.
2.Identification and genetic analysis of a novel goose astrovirus isolated from Jiangxi Province
Enfu HUANG ; Qianlang GU ; Yuxin LIU ; Jun CHEN ; Deping SONG ; Jiangnan HUANG ; Jia TAN ; Fanfan ZHANG
Chinese Journal of Veterinary Science 2025;45(11):2334-2342
In order to understand the genetic variation and whole genome characteristics of goose as-trovirus(GoAstV)in Jiangxi Province,a strain of GoAstV was successfully isolated from a typical gosling gout case sample and the whole genome sequencing and genetic characteristics of the isola-ted strain were analyzed.The results showed that the JXNC1 strain could be stably passaged on LMH cells and could cause mild cytopathic effects in LMH cells.Sequencing and analysis showed that the full-length genome of the strain was 7 173 bp,and its genetic relationship was the closest to the GZ2301(PP966939)reference strain,belonging to the GoAstV-Ⅱ genotype.The complete genome of JXNC1 strain shared 98.1%-98.8%nucleotide similarity with 22 GoAstV-Ⅱ reference strains,and the amino acid similarity of ORF2 gene was 97.6%-99.6%.At the same time,the analysis results showed that the mutation of the strain mainly occurred in the ORF2 gene,and there were 13 amino acid site mutations,of which T630I was a unique mutation.Animal regression experiments showed that the inoculation of JXNC1 strain could cause urate deposition in the or-gans of goslings,congestion and dilatation of hepatic sinusoids,and small focal necrosis of some hepatocytes.Renal tissue tubular dilatation,renal interstitial connective tissue hyperplasia.The re-sults of this study laid a foundation for accurate prevention and control of the disease.
3.Identification and genetic analysis of a novel goose astrovirus isolated from Jiangxi Province
Enfu HUANG ; Qianlang GU ; Yuxin LIU ; Jun CHEN ; Deping SONG ; Jiangnan HUANG ; Jia TAN ; Fanfan ZHANG
Chinese Journal of Veterinary Science 2025;45(11):2334-2342
In order to understand the genetic variation and whole genome characteristics of goose as-trovirus(GoAstV)in Jiangxi Province,a strain of GoAstV was successfully isolated from a typical gosling gout case sample and the whole genome sequencing and genetic characteristics of the isola-ted strain were analyzed.The results showed that the JXNC1 strain could be stably passaged on LMH cells and could cause mild cytopathic effects in LMH cells.Sequencing and analysis showed that the full-length genome of the strain was 7 173 bp,and its genetic relationship was the closest to the GZ2301(PP966939)reference strain,belonging to the GoAstV-Ⅱ genotype.The complete genome of JXNC1 strain shared 98.1%-98.8%nucleotide similarity with 22 GoAstV-Ⅱ reference strains,and the amino acid similarity of ORF2 gene was 97.6%-99.6%.At the same time,the analysis results showed that the mutation of the strain mainly occurred in the ORF2 gene,and there were 13 amino acid site mutations,of which T630I was a unique mutation.Animal regression experiments showed that the inoculation of JXNC1 strain could cause urate deposition in the or-gans of goslings,congestion and dilatation of hepatic sinusoids,and small focal necrosis of some hepatocytes.Renal tissue tubular dilatation,renal interstitial connective tissue hyperplasia.The re-sults of this study laid a foundation for accurate prevention and control of the disease.
4.Rapid construction of rPRV-ΔTK/EGFP variant strain using CRISPR/Cas9 sys-tem
Zaijiao YE ; Chuan ZENG ; Jun GU ; Peixia WANG ; Jinyan SHEN ; Deping SONG ; Dongyan HUANG ; Xiangdong WU ; Houjun HE ; Yuxin TANG ; Yu YE
Chinese Journal of Veterinary Science 2024;44(6):1223-1228
Pseudorabies virus(PRV)is the etiological agent of pseudorabies in pigs,which is char-acterized by dyspnea,reproductive disorders,and neurological diseases,and it spreads widely a-round the world.Since 2011,the newly emerged PRV variants have resulted in poor immunity pro-tection of traditional vaccine strains,and the original method of vaccine strain preparation is time-consuming and labor-intensive.Therefore,it is urgently needed to develop an efficient screening method of the vaccine strain at present.Using CRISPR/Cas9 gene editing technology in this study,two single guide RNAs(sgRNA)were designed targeting the virulence gene TK of PRV variant strain CH/JX/2016,and then the enhanced green fluorescent protein the reporter(EGFP)gene was inserted at the TK locus by a homologous repair plasmid.After multiple rounds of plaque puri-fication,the rPRV-ΔTK/EGFP strain was obtained.The results showed the cleavage efficiency of the two sgRNAs was extremely high.The preparation of rPRV-ΔTK/EGFP strain was succeed af-ter only three rounds of purification,and the EGFP expressed normally.The CRISPR/Cas9 system can edit the PRV gene simply,rapidly,and efficiently,and exhibits great potential in the construction of vaccine candidate strains.Meanwhile,the rescued rPRV-ΔTK/EGFP strain not only could be used as a tracer strain in PRV variant infection progresses,but also for subsequent antivi-ral drug screening.
5.Production and identification of monoclonal antibody against PDCoV N protein
Suxian LUO ; Nannan GUO ; Houjun HE ; Deping SONG ; Dongyan HUANG ; Yuxin TANG ; Yu YE
Chinese Journal of Veterinary Science 2024;44(12):2521-2525
Porcine deltacoronavirus(PDCoV)is one of the important pathogens associated with swine viral diarrhea,which results in enormous economic losses in the pig industry.Among the known encoding proteins,the nucleocapsid(N)protein is relatively conserved and serves as the dominant antigen of coronaviruses,making it an ideal candidate for the early and accurate diagnosis of infection.In this study,the PDCoV N protein was induced and expressed in a prokaryotic system using IPTG treatment at low temperatures.Six-week-old BALB/c mice were immunized with the recombinant PDCoV N protein purified by nickel column.Isolated spleen cells were harvested and fused with SP2/0 cells once the serum titers reached 1∶10 000,as determined by indirect ELISA.The positive monoclonal hybridoma with the highest titer of secretory antibodies were screened af-ter three rounds of limited dilution.Ultimately,indirect immunofluorescent assay(IFA)and West-ern blot were employed to confirm the characterization of the antibodies,and the subtypes of the heavy and light chains of the antibodies were determined.One strain of hybridoma against the PD-CoV N protein,designated as 2G12,was obtained,with a titer approaching 1∶40 960.The IFA as-say showed PDCoV was specifically bound to 2G12.Furthermore,the Western blot assay further showed PDCoV in the supernatant of infected cells or recombinant PDCoV N protein,reacted well with 2G12.The subtype of MAbs was determined as IgG1 with the light chains being κ.The MAbs generated against the PDCoV N protein in this study provide valuable support for the development of a diagnostic kit for PDCoV and play an important role in the functional research of the PDCoV N protein.
6.Production and identification of monoclonal antibody against PDCoV N protein
Suxian LUO ; Nannan GUO ; Houjun HE ; Deping SONG ; Dongyan HUANG ; Yuxin TANG ; Yu YE
Chinese Journal of Veterinary Science 2024;44(12):2521-2525
Porcine deltacoronavirus(PDCoV)is one of the important pathogens associated with swine viral diarrhea,which results in enormous economic losses in the pig industry.Among the known encoding proteins,the nucleocapsid(N)protein is relatively conserved and serves as the dominant antigen of coronaviruses,making it an ideal candidate for the early and accurate diagnosis of infection.In this study,the PDCoV N protein was induced and expressed in a prokaryotic system using IPTG treatment at low temperatures.Six-week-old BALB/c mice were immunized with the recombinant PDCoV N protein purified by nickel column.Isolated spleen cells were harvested and fused with SP2/0 cells once the serum titers reached 1∶10 000,as determined by indirect ELISA.The positive monoclonal hybridoma with the highest titer of secretory antibodies were screened af-ter three rounds of limited dilution.Ultimately,indirect immunofluorescent assay(IFA)and West-ern blot were employed to confirm the characterization of the antibodies,and the subtypes of the heavy and light chains of the antibodies were determined.One strain of hybridoma against the PD-CoV N protein,designated as 2G12,was obtained,with a titer approaching 1∶40 960.The IFA as-say showed PDCoV was specifically bound to 2G12.Furthermore,the Western blot assay further showed PDCoV in the supernatant of infected cells or recombinant PDCoV N protein,reacted well with 2G12.The subtype of MAbs was determined as IgG1 with the light chains being κ.The MAbs generated against the PDCoV N protein in this study provide valuable support for the development of a diagnostic kit for PDCoV and play an important role in the functional research of the PDCoV N protein.
7.Study on the application of standardized patient in practice courses of diagnostics
Wenjun PENG ; Feng MEI ; Yan LI ; Deping CAO ; Huilian YANG ; Shue FENG ; Shuaixian TAO ; Zihao SONG
Chinese Journal of Medical Education Research 2019;18(2):167-171
Objective To explore the application of standardized patients (SP) in practice course of diagnostics.Methods The two classes of 135 students majoring in clinical medicine were selected as research.After the practical course in inquiry,the questionnaires were used to compare the teaching effect between the traditional teaching and SP teaching.Meanwhile,the students evaluated and gave feedback on SP work.Results The questionnaires on teaching effect showed that 127 (94.8%) students believed that the SP teaching method could stimulate interest in inquiry,125 students (93.3%) thought it could increase the understanding and memory of the inquiry content.127 students (94.8%) thought it could improve the application ability of the inquiry method,128 students (95.5%) thought it could improve inquiry techniques,129 students (96.3%) thought it could enhance expression ability,89 students (66.4%) thought it could enhance cooperation with their classmates,117 students (87.3%) thought it could enhance the confidence in communication,115 students (85.8%) thought it could improve their ability in independent thinking and self-study,126 students (94.0%) thought it could improve the learning effect,117 (87.3%) students thought it could develop their clinical thinking ability,116 students (86.6%) thought it could improve the satisfaction to teaching form,111 students (82.8%) thought it could develop the ability to answer patients' problems in a manner which is easy for patients to understand,and 126 (94.0%) students thought it could improve the ability to interact with patients.All issues were significantly better than the traditional teaching method Compared with the traditional teaching method,the students' choices on various items have statistically significant differences (P<0.05).Conclusion Applying SP teaching to the practice courses of diagnostics is helpful to improve the teaching effect and cultivate students' clinical thinking ability.Thus,it is a promising teaching method in medical undergraduate education and worthy of further discussion and popularization.

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