1.Prokaryotic expression and preliminary analysis of immunogenicity of outer mem-brane protein of yak-derived Escherichia coli OmpA
Shinan ZHANG ; Shengyi HAN ; Tian SHI ; Shuping LI ; Guoyuan HU ; Rui GAO ; Jiaqi TIAN ; Wenwen ZHOU ; Shengqing LI
Chinese Journal of Veterinary Science 2025;45(3):458-465,472
The amino acid sequences of the OmpA protein isolated from Escherichia coli QML2206-1(E.coli QML2206-1)in our laboratory were analyzed for homology with different strains of OmpA proteins using bioinformatics software,and the OmpA protein was analyzed for its physicochemical properties,transmembrane structure and signal peptide prediction,B-cell anti-genic epitope prediction,secondary and tertiary structure prediction.The OmpA gene fragment was ligated with pET-32a vector to construct a prokaryotic expression vector,which was purified by a nickel column affinity purification system after prokaryotic expression and optimization of ex-pression conditions in BL21(DE3).The purified recombinant protein was fully mixed with Freund's adjuvant to immunize mice,and the levels of mouse-specific IgG antibody and the expression levels of cytokines CD4,CD8 and IL-4 in mouse serum were detected by ELISA,and the immuno-protective effect was evaluated by mouse attack protection test.OmpA protein is a hydrophilic protein with no transmembrane structural domains and a secondary structure consisting mainly of irregular coils(47.98%)and α-helices(29.77%),with 12 antigenic epitopes that can bind to anti-bodies produced by B cells.The recombinant protein OmpA with a relative molecular mass of a-bout 55 kDa was successfully obtained by prokaryotic expression,and the highest expression was induced by IPTG concentration of 0.000 4 mmol/L for 6 h at 37 ℃.The serum-specific IgG anti-body potency of recombinant protein immunized mice was up to 1∶32 000;the expression levels of CD4,CD8 and IL-4 in the serum of immunized mice were elevated compared with those of the con-trol group.The survival rate of mice was 80%and 40%after attack with minimum lethal dose(MLD)and 2 times minimum lethal dose(2MLD),respectively.OmpA recombinant protein has good antigenicity and certain immunoprotective effects,and this study provides a technical basis for the next step in the development of a genetically engineered subunit vaccine against yak-appli-cable E.coli based on OmpA protein.
2.Prokaryotic expression and preliminary analysis of immunogenicity of outer mem-brane protein of yak-derived Escherichia coli OmpA
Shinan ZHANG ; Shengyi HAN ; Tian SHI ; Shuping LI ; Guoyuan HU ; Rui GAO ; Jiaqi TIAN ; Wenwen ZHOU ; Shengqing LI
Chinese Journal of Veterinary Science 2025;45(3):458-465,472
The amino acid sequences of the OmpA protein isolated from Escherichia coli QML2206-1(E.coli QML2206-1)in our laboratory were analyzed for homology with different strains of OmpA proteins using bioinformatics software,and the OmpA protein was analyzed for its physicochemical properties,transmembrane structure and signal peptide prediction,B-cell anti-genic epitope prediction,secondary and tertiary structure prediction.The OmpA gene fragment was ligated with pET-32a vector to construct a prokaryotic expression vector,which was purified by a nickel column affinity purification system after prokaryotic expression and optimization of ex-pression conditions in BL21(DE3).The purified recombinant protein was fully mixed with Freund's adjuvant to immunize mice,and the levels of mouse-specific IgG antibody and the expression levels of cytokines CD4,CD8 and IL-4 in mouse serum were detected by ELISA,and the immuno-protective effect was evaluated by mouse attack protection test.OmpA protein is a hydrophilic protein with no transmembrane structural domains and a secondary structure consisting mainly of irregular coils(47.98%)and α-helices(29.77%),with 12 antigenic epitopes that can bind to anti-bodies produced by B cells.The recombinant protein OmpA with a relative molecular mass of a-bout 55 kDa was successfully obtained by prokaryotic expression,and the highest expression was induced by IPTG concentration of 0.000 4 mmol/L for 6 h at 37 ℃.The serum-specific IgG anti-body potency of recombinant protein immunized mice was up to 1∶32 000;the expression levels of CD4,CD8 and IL-4 in the serum of immunized mice were elevated compared with those of the con-trol group.The survival rate of mice was 80%and 40%after attack with minimum lethal dose(MLD)and 2 times minimum lethal dose(2MLD),respectively.OmpA recombinant protein has good antigenicity and certain immunoprotective effects,and this study provides a technical basis for the next step in the development of a genetically engineered subunit vaccine against yak-appli-cable E.coli based on OmpA protein.
3.An exploratory study of the domestic robotic surgical system for general surgery
Jing YANG ; Zeping ZHANG ; Guoyuan YANG ; Yiyun PENG ; Meijuan SONG ; Weipeng ZHAN ; Hongwei TIAN ; Ming HU ; Changfeng MIAO ; Dongdong CHEN ; Wei FANG ; Wutang JING ; Yuntao MA
Chinese Journal of General Surgery 2024;39(9):698-702
Objective:To evaluate the safety and efficacy of the domestic robotic surgical system for general surgery.Methods:A prospective single-center, single-arm exploratory study was conducted at Gansu Provincial People's Hospital from Jun 2022 to Oct 2023, enrolling 54 patients undergoing general surgery using domestically produced Toumai? Endoscopic Surgical Robotic System. The primary study endpoint was surgical success rate, and the secondary study endpoints were intraoperative bleeding, operative time, complications, system performance, hospitalization days.Results:In this study, robotic surgery was successfully completed in 52 patients, and in 2 patients undergoing thyroid operation it was converted to open surgery due to bleeding, with a success rate of 96%, no organ injury or death during surgery, and no system failure. The types of surgery included cholecystectomy, radical gastric cancer resection, radical colorectal cancer resection, inguinal hernia repair, partial hepatectomy, total thyroidectomy and choledocho-jejunal anastomosis.Conclusion:The study provides preliminary evidence of the safety and efficacy of the Toumai? Endoscopic Surgical Robotic System for the treatment of general surgical diseases.
4.Drug Resistance and Prognosis of 150 Cases of Peritoneal Dialysis-associated Peritonitis
Yueyuan WU ; Xiaohua DAI ; Jie XU ; Xianfeng ZHANG ; Deyu XU ; Kun HU ; Lei SHEN ; Guoyuan LU ; Qiang HAN ; Yongfu HANG
Herald of Medicine 2024;43(2):287-291
Objective To analyze the pathogenic bacteria and drug resistance of peritoneal dialysis-associated peritonitis(PDAP),and provide a clinical reference for the rational use of antibiotics.Methods The demographic data of PDAP patients admitted to the peritoneal dialysis(PD)Center of the First Affiliated Hospital of Soochow University from July 1,2015 to December 30,2021 were collected,and the pathogens,drug resistance and prognosis were retrospectively analyzed.Results A total of 150 episodes of PDAP occurred in 92 patients.The positive rate of PD fluid culture was 61.33%,including 65 cases(70.65%)of Gram-positive(G+)bacteria,mainly Staphylococcus and Streptococcus.Gram-negative(G-)bacteria were in 16 cases(17.39%),mainly Escherichia coli and Enterobacter cloacae.There were 11 cases(11.96%)of multiple infections,including 5 cases of combined fungal infection.From 2016 to 2021,the incidence of G+bacteria-related PDAP decreased from 14 to 8 cases.G+strains were resistant to methicillin(35.00%),and were sensitive to linezolid(100.00%),teicoplanin(100.00%)and rifampicin(100.00%).The sensitivity rate to vancomycin was 98.59%.G-strains were sensitive to ceftazidime(86.36%),ceftizoxime(88.89%)and amikacin(100.00%).The MIC of vancomycin against Staphylococcus showed an upward trend in 2019-2021.The overall cure rate of PDAP was 81.33%in patients who responded to antibiotic treatment,and the cure rate of G+bacteria was higher than that of multiple infections(89.23%vs.36.36%,P<0.01).The outcome of patients with multiple infections,especially those with concurrent fungal infection was poor.Conclusion The incidence of PDAP in the PD center has shown a decreasing trend in recent years.G+bacteria are still the main pathogenic bacteria causing PDAP,and they are highly resistant to methicillin,so vancomycin should be used as empirical therapy.For G-bacteria,cefotaxime and amikacin can be chosen as empirical therapy.There is a drift in the MIC values of vancomycin against Staphylococcus in the study period,so it is necessary to monitor the MIC of vancomycin against Staphylococcus and its changing trend.
5.The application of anchor-paired classroom combined case teaching mode in the teaching of pediatric surgery
Guoyuan LI ; Fang HU ; Peiying LIU ; Ru WEI ; Jiahang ZENG ; Gaoyan DENG ; Le LI
Modern Hospital 2024;24(12):1962-1965
Objective To explore the application effect of the anchored and paired classroom combined case teaching mode in the teaching of pediatric Surgery.Methods The five-year undergraduate students of pediatrics in Guangzhou Medical U-niversity from September 2022 to June 2024 were selected as the research objects.Traditional teaching mode was used as the con-trol group,and the anchored and paired classroom combined case teaching mode was used as the research group.According to the"Teaching Evaluation Form",the learning process,learning effect,classroom atmosphere and teaching strategies were evaluated,and the test results of the two groups of medical students were compared,so as to evaluate the degree of students'mastery of bas-ic knowledge and clinical thinking ability.Results The study group scored higher than the control group,and the difference was statistically significant(P<0.05).Conclusion The anchored and paired classroom combined case teaching mode can improve the teaching quality of Pediatric Surgery,improve the classroom atmosphere,and enhance the learning interest and autonomy of medical students.
6.The application of anchor-paired classroom combined case teaching mode in the teaching of pediatric surgery
Guoyuan LI ; Fang HU ; Peiying LIU ; Ru WEI ; Jiahang ZENG ; Gaoyan DENG ; Le LI
Modern Hospital 2024;24(12):1962-1965
Objective To explore the application effect of the anchored and paired classroom combined case teaching mode in the teaching of pediatric Surgery.Methods The five-year undergraduate students of pediatrics in Guangzhou Medical U-niversity from September 2022 to June 2024 were selected as the research objects.Traditional teaching mode was used as the con-trol group,and the anchored and paired classroom combined case teaching mode was used as the research group.According to the"Teaching Evaluation Form",the learning process,learning effect,classroom atmosphere and teaching strategies were evaluated,and the test results of the two groups of medical students were compared,so as to evaluate the degree of students'mastery of bas-ic knowledge and clinical thinking ability.Results The study group scored higher than the control group,and the difference was statistically significant(P<0.05).Conclusion The anchored and paired classroom combined case teaching mode can improve the teaching quality of Pediatric Surgery,improve the classroom atmosphere,and enhance the learning interest and autonomy of medical students.
7.Role of immunoglobulin glycosylation in autoimmune diseases
Xiaoli ZENG ; Guoyuan ZHANG ; Chaojun HU
Chinese Journal of Laboratory Medicine 2021;44(9):868-872
Glycosylation is one of the most important post-translational modification in protein biosynthesis. Immunoglobulin glycosylation can exert anti-inflammatory or pro-inflammatory activity by regulating antibody stability and affecting its interaction with different FcγRs. In recent years, a large number of studies have confirmed that abnormal glycosylation of immunoglobulin plays a key role in the pathogenesis of autoimmune diseases. Its products, such as sugar chain structure or/and glycosylated proteins, can be detected by lectin microarray and other technologies, are expected to become new serum markers of autoimmune diseases. It has a broad application prospect in disease diagnosis, condition monitoring, prognosis evaluation and treatment.
8.Latest advances in the pathogenesis of hepatogenous diabetes
Han HU ; Caiyun TIAN ; Guoyuan ZHANG ; Shide LIN
Journal of Clinical Hepatology 2021;37(2):429-432
Hepatogenous diabetes (HD) is a common complication of end-stage liver disease, and many studies have confirmed its adverse effect on prognosis. In recent ten years, a great number of studies have been conducted on the pathogenesis of HD and some progress has been made. This article reviews the research advances in the pathogenesis of HD, in order to provide a reference for the diagnosis and treatment of HD by clinicians.
9.Influence of glucose metabolism disorder on complications associated with liver cirrhosis
Caiyun TIAN ; Han HU ; Guoyuan ZHANG ; Shide LIN
Journal of Clinical Hepatology 2021;37(5):1197-1200.
Both diabetes mellitus and liver cirrhosis have high incidence rate and mortality rate around the world, and in recent ten years, scholars in China and globally have conducted many studies on the association between diabetes mellitus and liver cirrhosis. This article systematically reviews the advances in the basic and clinical research on the influence of diabetes mellitus on liver cirrhosis and its complications and summarizes possible mechanisms. The results show that diabetes mellitus can accelerate the process of liver fibrosis, increase the risk of complications and progression to liver cancer in patients with liver cirrhosis, and reduce their survival rate.
10.Variation and clinical significance of circulation annexin Ⅱ in patients with systemic lupus erythematosus
Kun HU ; Guoyuan LU ; Miao JIANG ; Zhaoyue WANG
Chinese Journal of Rheumatology 2011;15(5):332-335
Objective To compare the level of Annexin Ⅱ in patients with systemic lupus erythematosus(SLE),diabetic nephropathy(DN),chronic glomerulonephritis and normal controls,and explorle the significance of the annexin Ⅱ in SLE.Methods Thirty-five cases of patients with SLE,ten cases of patients with DN,ten cases of patients with chronic glomerulonephritis were enrolled in this study,twenty cases of healthy controls were also enrolled.Circulating annexin Ⅱ in white blood cells was detected bv flow cytometry.Student's t test,variance analysis and Lineat correlation analysis were used for statistial analysis.Resuits Compared with healthy controls,the level of annexin Ⅱ in white blood cells in SLE patients (7.1±2.9)%and DN patients(8.0±3.7)%were significantly lower than that of the healthy controls(P<0.01,p<0.05).In the SLE group,the level of annexin Ⅱ of patients who had more active disease(SLEDAI≥9)decreased more thall those with less active disease(SLEDAI<9),(P<0.05).A positive correlation was found between annexin Ⅱ and serum albumin level(r:0.439,P<0.01),but negative correlation was found between annexin and urine protein/urine creatinine(r=-0.382,P<0.05),SLEDAI(r=-0.417,P<0.05),D-dimer(r=-0.336.p<0.05) levels.Conclusion The level of annexin Ⅱ is decreased in patients with SLE,so it can renectthe abnormality of coagulation and fibrinolytic systems,and it may be used as a good indicator for prothrombotic status in SLE patients.It can be helpful to evaluatethe activity of the disease and the therapeutic efficacy.

Result Analysis
Print
Save
E-mail