1.Isolation,identification and pathogenicity analysis of two avain infectious bronchi-tis virus strain
Hanwen ZHANG ; Yaru GAO ; Yang WANG ; Yapeng SONG ; Wenming GAO ; Lin LIU ; Xiaoyang CAO ; Jingrui LIU ; Xinsheng LI
Chinese Journal of Veterinary Science 2025;45(10):2133-2141,2155
This study aims to investigate the isolate and identify of infectious bronchitis virus(IBV)in chickens,and study its genetic variation and pathogenicity.In 2023,two strains named CK/CH/HN/SQ202301 and CK/CH/HN/SQ202302 were obtained from suspected infectious bronchitis(IB)infected materials collected in a region of Henan Province,China.Further analysis showed that the two isolates belong to the G Ⅰ-13 and GⅥ genotypes,respectively.The cleavage sites of S protein were all RRSRR.The prediction of glycosylation sites showed that the two isolates had 18 and 12 N-glycosylation sites respectively,but no O-glycosylation site.Recombinant analysis shows that C2023-1 was a recombinant strain.Pathogenicity was assessed by infecting 1-day-old SPF chicks with the two isolates,and the results showed that C2023-1 strain infection could cause clini-cal symptoms such as depression and head shaking,as well as death in chicks,with a mortality rate of 37.5%.There were no clinical symptoms or deaths after infection with C2023-2 strain.Viral load test results showed that both isolates continued to detoxify until the 10th day,and had strong rep-lication capacity in the kidney,trachea and bursa of Fabricius.The results indicate significant differences in the genetic characteristics and pathogenicity of the two isolates due to their different genotypes.This study not only provides new epidemiological data on IB,which contributes to a bet-ter understanding of IBV's epidemiological features and control challenges,but also adds valuable bioinformatics resources for IBV by analyzing its variation mechanisms and biological information.
2.Expression and immunogenicity study of IBV S1 fusion protein containing FAdV-4 Fiber 2 B cell epitopes
Yaru GAO ; Yang WANG ; Hanwen ZHANG ; Lin LIU ; Xiaoyang CAO ; Jingrui LIU ; Bingjie LI ; Jingjing CHANG ; Yapeng SONG ; Wenming GAO ; Xinsheng LI
Chinese Journal of Veterinary Science 2025;45(8):1616-1623
To verify whether the two B-cell epitopes Pep1 and Pep4 in the FAdV-4 WZ fiber can be used as candidate epitopes for multivalent epitope vaccines,epitopes Pep1 and Pep4 were tandemly linked with the chicken infectious bronchitis virus strain M41 S1 protein gene in different patterns,and a recombinant fusion plasmid was constructed and expressed in E.coli BL21(DE3).It was confirmed by Western blot and ELISA tests that all four expressed fusion proteins reacted specific-ally with anti-M41 whole virus serum and WZ strain anti-Fiber 2-knob protein serum.After purifi-cation and immunization of BALB/c mice,specific antibodies against the peptide epitopes were de-tected in mouse sera.The results showed that the Pep4 epitope induced a stronger immune re-sponse than the Pep1 epitope.When Pep1 was connected with the amino and carboxyl termini of the fusion protein,respectively,both resulted in the production of the same level of anti-Pep1 anti-bodies in the immunized animals,whereas when Pep4 was connected with the carboxyl terminus of the fusion protein,the immunized animals produced a higher level of anti-Pep4-specific antibodies.This research indicates that the B cell epitopes Pep1 and Pep4 of the reactive WZ strain Fiber 2,when conjugated with proteins to form fusion proteins,can enhance the immunogenicity of Pep1 and Pep4 without affecting the antigenicity of the carrier protein.This study provides technical support and serves as a reference for the design and development of a multivalent epitope vaccine for FAdV-4.
3.Expression and immunogenicity study of IBV S1 fusion protein containing FAdV-4 Fiber 2 B cell epitopes
Yaru GAO ; Yang WANG ; Hanwen ZHANG ; Lin LIU ; Xiaoyang CAO ; Jingrui LIU ; Bingjie LI ; Jingjing CHANG ; Yapeng SONG ; Wenming GAO ; Xinsheng LI
Chinese Journal of Veterinary Science 2025;45(8):1616-1623
To verify whether the two B-cell epitopes Pep1 and Pep4 in the FAdV-4 WZ fiber can be used as candidate epitopes for multivalent epitope vaccines,epitopes Pep1 and Pep4 were tandemly linked with the chicken infectious bronchitis virus strain M41 S1 protein gene in different patterns,and a recombinant fusion plasmid was constructed and expressed in E.coli BL21(DE3).It was confirmed by Western blot and ELISA tests that all four expressed fusion proteins reacted specific-ally with anti-M41 whole virus serum and WZ strain anti-Fiber 2-knob protein serum.After purifi-cation and immunization of BALB/c mice,specific antibodies against the peptide epitopes were de-tected in mouse sera.The results showed that the Pep4 epitope induced a stronger immune re-sponse than the Pep1 epitope.When Pep1 was connected with the amino and carboxyl termini of the fusion protein,respectively,both resulted in the production of the same level of anti-Pep1 anti-bodies in the immunized animals,whereas when Pep4 was connected with the carboxyl terminus of the fusion protein,the immunized animals produced a higher level of anti-Pep4-specific antibodies.This research indicates that the B cell epitopes Pep1 and Pep4 of the reactive WZ strain Fiber 2,when conjugated with proteins to form fusion proteins,can enhance the immunogenicity of Pep1 and Pep4 without affecting the antigenicity of the carrier protein.This study provides technical support and serves as a reference for the design and development of a multivalent epitope vaccine for FAdV-4.
4.Isolation,identification and pathogenicity analysis of two avain infectious bronchi-tis virus strain
Hanwen ZHANG ; Yaru GAO ; Yang WANG ; Yapeng SONG ; Wenming GAO ; Lin LIU ; Xiaoyang CAO ; Jingrui LIU ; Xinsheng LI
Chinese Journal of Veterinary Science 2025;45(10):2133-2141,2155
This study aims to investigate the isolate and identify of infectious bronchitis virus(IBV)in chickens,and study its genetic variation and pathogenicity.In 2023,two strains named CK/CH/HN/SQ202301 and CK/CH/HN/SQ202302 were obtained from suspected infectious bronchitis(IB)infected materials collected in a region of Henan Province,China.Further analysis showed that the two isolates belong to the G Ⅰ-13 and GⅥ genotypes,respectively.The cleavage sites of S protein were all RRSRR.The prediction of glycosylation sites showed that the two isolates had 18 and 12 N-glycosylation sites respectively,but no O-glycosylation site.Recombinant analysis shows that C2023-1 was a recombinant strain.Pathogenicity was assessed by infecting 1-day-old SPF chicks with the two isolates,and the results showed that C2023-1 strain infection could cause clini-cal symptoms such as depression and head shaking,as well as death in chicks,with a mortality rate of 37.5%.There were no clinical symptoms or deaths after infection with C2023-2 strain.Viral load test results showed that both isolates continued to detoxify until the 10th day,and had strong rep-lication capacity in the kidney,trachea and bursa of Fabricius.The results indicate significant differences in the genetic characteristics and pathogenicity of the two isolates due to their different genotypes.This study not only provides new epidemiological data on IB,which contributes to a bet-ter understanding of IBV's epidemiological features and control challenges,but also adds valuable bioinformatics resources for IBV by analyzing its variation mechanisms and biological information.
5.Research Progress on Chemical Constituents of Alpinia oxyphylla and its Pharmacological Activities
Haibo LI ; Mi ZHOU ; Jie DONG ; Zhenzhong WANG ; Liang CAO ; Xinsheng YAO ; Yang YU ; Wei XIAO
World Science and Technology-Modernization of Traditional Chinese Medicine 2024;26(7):1870-1887
As a commonly used traditional Chinese medicine,Alpinia oxyphylla is widely used as both medicine and edible resources.A.oxyphylla has the effects of warming the kidney,consolidating essence,contracting urine,warming the spleen,stopping diarrhea and absorbing saliva,which mainly treated diseases caused by kidney deficiency and spleen cold.A.oxyphylla is rich in chemical components,mainly including 194 volatile oil,121 terpenoids(including 111 sesquiterpenoids),19 diphenylheptanes,ten flavonoids,ten bases and nucleosides,four steroids,eight glycosides and 13 organic acids.It has a wide range of pharmacological effects such as anti-AD/PD,anti-tumor,anti-inflammatory,antioxidant,etc.This article reviews the chemical components and pharmacological effects of A.oxyphylla,in order to provide reference for its further development and rational application.
6.Real-time evaluation of pulmonary ventilation of HP movements based on electrical impedance tomography:an observational study
Zhijun GAO ; Hang WANG ; Chunchen WANG ; Yilong QIN ; Xinsheng CAO ; Lin YANG
Journal of Army Medical University 2024;46(16):1882-1889,1896
Objective To explore the feasibility of electrical impedance tomography(EIT)for real-time and accurate monitoring of respiration during HP anti-G movements and the key parameters of pulmonary ventilation.Methods Twelve healthy male students in our university were enrolled in September 2023 and subjected in this study.HP anti-G movements were performed 3 times each for 30 s in the anti-G physiological training apparatus,during which EIT and ophthalmic horizontal arterial pressure were measured to analyze the relationship of the global and local parameters of pulmonary ventilation,including inspiratory volume(IV),expiratory uniform(EU),expiratory speed(ES),center of ventilation(COV)and right-to-left lung ventilation ratio(RtoL)with anti-G ability of anti-G straining maneuver(AGSM).Results The average eye horizontal systolic blood pressure(SBP)at the eye level was 148.82±22.75 mmHg during HP anti-G movements,which was significantly higher than that during quiet breathing(PJ)(95.17±8.51 mmHg,P<0.001).From the global pulmonary ventilation,the participants had significantly increased IV during HP anti-G movements(P<0.001).According the d value(mean increase of eye horizontal SBP=SBPHP-SBPPJ),the subjects were divided into 3 groups,with the d value of>60,30~60 and<30 mmHg,respectively.The inspiratory volume ratio(IVHP/IVPJ)was the highest in the>60 mmHg group and the smallest in the<30 mmHg group(P<0.01).The subjects had significantly decreased EU and more evenly expiration(P<0.05),but no change was seen in the expiratory uniformity ratio(EUHP/EUPJ)among the 3 groups.ES was obviously faster during HP anti-G movements(P<0.001),and the expiratory speed ratio(ESHP/ESPJ)had no significant difference among the 3 groups.The inspiratory time and expiratory time were 0.77±0.32 and 1.59±0.21 s,respectively,and both of them were notably shorter during HP anti-G movements(P<0.001,P<0.01).From the local pulmonary ventilation,COV during HP anti-G movements was significantly smaller than that during PJ(P<0.001),and the ventilation center deviated to the ventral side,and RtoL was decreased and the ventilation distribution deviated to the left lung(P<0.05).Conclusion EIT can perform real-time imaging of pulmonary global and local ventilation during HP anti-G movements,and it has a great application prospect in AGSM training and monitoring.
7.Curative effect analysis of transplantation in patients with myelodysplastic syndrome with ASXL1 gene mutation
Yupei ZHANG ; Xinsheng XIE ; Yajie SHI ; Weijie CAO ; Rong GUO ; Dingming WAN
The Journal of Practical Medicine 2023;39(23):3087-3092
Objective To investigate the efficacy and influencing factors of allo-HSCT in the treatment of MDS patients with ASXL1+.Methods The second-generation sequencing technique was used to detect 22 gene mutations in 247 newly diagnosed MDS patients in our hospital.The patients were divided into chemotherapy group and transplant group according to treatment style.The differences of OS and PFS between the two groups were compared,and the influencing factors of prognosis of transplant patients were analyzed.Results ASXL1+ was detected in 75 patients(30.36%),with a median mutation ratio of 42.93(18.10,58.39)%,10 received supportive treatment,43 received demethylation therapy or demethylation combined with pre-excitation therapy,and 22 received allo-HSCT.2-year PFS rate and OS rate of transplantation group were significantly higher than that of chemotherapy group(P<0.05).The 2-year OS rate in the low ASXL1 mutation load group(VAF≤42.93%)was significantly higher than that in the high ASXL1 mutation load group(VAF>42.93%)(P<0.05).In the context of allo-HSCT in patients with ASXL1+,2-year OS and PFS rates were significantly reduced in patients with RUNX1+ or ASXL1+(P<0.05);Multivariate analysis showed that high mutation load of ASXL1 or U2AF1+ were independent risk factors for OS in transplant patient(P<0.05).U2AF1+ were the risk factors for PFS(P<0.05).Conclusion allo-HSCT significantly improved the prognosis of patients with ASXL1+ MDS.High ASXL1 mutation load or U2AF1+ were independent risk factors affecting the outcome of allo-HSCT.
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.Efficacy comparison of the second generation TKI combined with allogeneic hematopoietic stem cell transplantation or chemotherapy in the treatmeat of Philadelphia chromosome-positive acute lymphoblastic leukemia
Suping ZHANG ; Dingming WAN ; Zhongxing JIANG ; Weijie CAO ; Xue LI ; Li LI ; Xinsheng XIE ; Rong GUO ; Yanfang LIU ; Ling SUN
Chinese Journal of Organ Transplantation 2020;41(6):346-350
Objective:To compare the efficacy of the second generation tyrosine kinase inhibitor dasatinib combined with allogeneic hematopoietic stem cell transplantation(allo-HSCT)or chemotherapy in the treatment of Ph + acute lymphoblastic leukemia (Ph + ALL). Methods:A total of 56 Ph + ALL patients received dasatinib from January 2014 to June 2018. According to whether or not allo-HSCT was performed, they were divided into transplantation group(n=22)and chemotherapy group(n=34). The total survival rate(OS), disease-free survival rate(DFS), relapse and non-recurrence mortality(NRM)were compared between two groups. Results:The 2-year OS, DFS and cumulative recurrence rates were 69.1 % vs 47.8 %, 62.2 % vs 43.1 % and 14.6 % vs 44.1 % in transplantation and chemotherapy groups respectively. Significant inter-group differences existed in 2-year DFS, DFS and cumulative recurrence rates. The value of NRM was higher in transplantation group than that in chemotherapy group(18.6 % vs 14.1 %). However, the difference was statistically insignificant( P=0.476). Conclusions:The efficacy of dasatinib plus allo-HSCT is superior to that of dasatinib plus chemotherapy in the treatment of Ph + ALL.
10.Biosynthesis of clinically used antibiotic fusidic acid and identification of two short-chain dehydrogenase/reductases with converse stereoselectivity.
Zhiqin CAO ; Shaoyang LI ; Jianming LV ; Hao GAO ; Guodong CHEN ; Takayoshi AWAKAWA ; Ikuro ABE ; Xinsheng YAO ; Dan HU
Acta Pharmaceutica Sinica B 2019;9(2):433-442
Fusidic acid is the only fusidane-type antibiotic that has been clinically used. However, biosynthesis of this important molecule in fungi is poorly understood. We have recently elucidated the biosynthesis of fusidane-type antibiotic helvolic acid, which provides us with clues to identify a possible gene cluster for fusidic acid ( cluster). This gene cluster consists of eight genes, among which six are conserved in the helvolic acid gene cluster except and . Introduction of the two genes into the NSAR1 expressing the conserved six genes led to the production of fusidic acid. A stepwise introduction of and revealed that the two genes worked independently without a strict reaction order. Notably, we identified two short-chain dehydrogenase/reductase genes and in the cluster, which showed converse stereoselectivity in 3-ketoreduction. This is the first report on the biosynthesis and heterologous expression of fusidic acid.

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