1.Synthesis of peptide of BALF4 gene of Epstein-Barr virus and application in the detection of nasopharyngeal carcinoma
Jianhua HUANG ; Yuan LIAO ; Jiangling CHEN ; Bo HU
The Journal of Practical Medicine 2016;32(19):3187-3189
Objective To analyze the application of recombinant gp125 proteins in serological diagnosis of nasopharyngeal carcinoma (NPC). Methods B cell epitopes of EBV gp125 protein were forecasted by internet server and the peptides were synthesized and coated to microplate for the detection of EBV-IgA antibody in NPC patients and healthy donors by ELISA. Results Six peptides were synthesized, wherein two showed low specificity and others displayed significantly higher positive rate in the NPC group than in the healthy group (P <0.001). Conclusion The BALF4 gene peptides are successfully synthesized and epitopes with high specificity and strong immunogenicity are screened for the preparation of gp125 antigen with high specificity and sensitivity.
2.Analysis of the main organs, hematological and biochemical indexes in congenital cataract mice
Jiangling YUAN ; Xiaohui XU ; Yan ZHANG ; Ruifang HUANG ; Jin XIONG ; Xinru CHEN
Chinese Journal of Comparative Medicine 2016;26(6):32-36
Objective To establish the baseline data of body weight, main organ weights, hematological and biochemical indexes in SPF congenital cataract mice. Methods Body weight, main organs weights, hematological and biochemical indexes of the congenital cataract mice were determined at 28 days and 56 days of age, respectively. Normal KM mice in the same age were taken as control. Results There were no statistically significant differences in all indexes of the mice at 28 days of age. Compared with the 56?day old normal KM mice: (1) Statistically significant differences were found in the body weight, and weights of the heart, liver, spleen, lung, kidney and testis ( P<0?05 or P<0?01; ( 2 ) Statistically significant differences were found in hematological indexes WBC, PLT, MPV, LYMP, PDW for female mice and MPV, PDW for male mice (P<0?01);(3) Among the biochemical indexes, there were also statistically significant differences in UREA, ALP, TP, UA, TG, GLU for female and ALT, ALP, TP, ALB, UA, GLU for male mice. Conclusions There are statistical differences in the body weight, main organ weights, hematological indexes and biochemical indexes between the congenital cataract mice and normal KM mice at 56 days of age. These results may provide a useful reference for future research.
3.Genomic epitopes of Xinjiang hemorrhagic fever virus and the evaluation of its laboratory testing value
Huahua ZHANG ; Guoyu ZHAO ; Yang LI ; Shen SHI ; Ran AN ; Jiangling YUAN ; Dongliang LIU ; Jingrui DOU ; Tao LUO ; Surong SUN ; Yujiang ZHANG
Chinese Journal of Laboratory Medicine 2023;46(2):127-136
Objective:To grasp the distribution of fine antigenic epitope profiles of nucleoprotein (NP) and glycoprotein (GP) fragments of Crimean-Congo hemorrhagic fever virus (CCHFV) and to clarify the value of dominant antigenic epitopes in laboratory testing of Crimean-Congo hemorrhagic fever (CCHF).Methods:In a minimal synthetic short peptide consisting of 8 amino acids was segmentally expressed by CCHFV YL04057 strain using a modified bio-peptide synthesis method from 2014 to 2021 in the laboratory of Xinjiang University, College of Life Sciences. Using CCHFV polyclonal antibody or monoclonal antibody 14B7 (IgM) or CCHFV-positive sheep serum as antibodies, the minimal antigenic epitopes (BCEs) with antigenic activity on NP and GP fragments were identified by immunoblotting, and the obtained BCEs with sequence polymorphism were spatially clustered with CCHFV from different regions using the neighbor-joining method to determine the combination mode of BCEs with geographical correlation of regional distribution, to explore its application in establishing serological diagnosis. A prokaryotic expression plasmid (pET-32a), an E. coli expression plasmid (pGEX-KG) and a prokaryotic expression plasmid with an incomplete glutathione (GST188) tag (pXXGST-ST-1) were used to construct and express six dominant antigenic epitopes of different peptide lengths on NP fragments, and an indirect Enzyme-linked immunosorbent assay (ELISA) was established. CCHF sheep serum identified by immunofluorescence assay (IFA) was used as a control, and the specificity, sensitivity and overall compliance of the recombinant proteins with different peptide lengths of antigenic epitopes with IFA assay results were statistically analyzed. Results:CCHFV, NP and GP fragments had a total of 30 antigenically active BCEs, among which the core intermediate fragment NP2 (aa 170 th-305 th), which had a concentration of antigenic epitopes in the NP fragment, has 6 BCEs, and the NP1 (aa 1 st-200 th) and NP3 (aa 286 th-482 nd) at both ends have 9 BCEs; the Gc (aa 1 st-558 th) and Gn (aa 533 th-708 th) fragments of the GP fragment have 14 BCEs and a long antigenic peptide (AP) containing 15 amino acids, and the amino acid sequence homology of the NP fragment BCEs was 97.1% and that of the GP fragment BCEs was 89.1%. There was a significant difference ( P=0.0281, P<0.05). Among the 9 BCEs with sequence polymorphism in the GP fragment, 6 combined BCEs from GnEc1, GnE2, GnE4, GcE3, GcE6 and GcAP-4 (Ap) could cluster 15 CCHFV strains from different regions of the world into 5 geographical taxa, AsiaⅠ, AsiaⅡ, AficaⅠ, AficaⅡ and Europe. The constructs expressing PET-32a-NP (full length), PGEX-KG-NP2 (aa 170 th-305 th), pGEX-KG-NP2-1 (aa 235 th-275 th), PGEX-KG-NP2-1-1 (aa 237 th-256 th), pXXGST-1-NP2-1-2 (aa 250 th-265 th) and PGEX KG-NP2-1-3 (aa 260 th-276 th), six recombinant proteins CCHFV NP rabbit polyclonal antiserum (pAb) Western Blotting reaction positive, 33 sheep sera tested by IFA XHF as a reference, the sensitivity of the assay established by indirect ELISA using the recombinant proteins constructed from two fragments of NP2 and NP2-1 as antigens. The sensitivity, specificity and overall compliance were the best, with 73.4% (11/15) and 66.7% (10/15) for sensitivity, 100% (18/18) and 94.4% (17/18) for specificity, and 87.9% (29/33) and 81.8% (27/33) for overall compliance. Conclusion:CCHFV NP and GP are distributed with a high number of BCEs with antigenic immunoreactivity, among which the dominant antigenic epitopes are of high value in the laboratory serological diagnosis of CCHF.
4.Molluscicidal effect of spraying different formulations of niclosamide ethanolamine salt with drones against Oncomelania hupensis in ditches
Zelin ZHU ; Xia ZHANG ; Junyi HE ; Ying CHEN ; Weisi WANG ; Hehua HU ; Chunli CAO ; Ziping BAO ; Suying GUO ; Liping DUAN ; Yi YUAN ; Jing XU ; Shizhu LI ; Xiaonong ZHOU
Chinese Journal of Schistosomiasis Control 2024;36(5):527-530
Objective To evaluate the molluscicidal effect of spraying different formulations of niclosamide ethanolamine salt with drones against Oncomelania hupensis in ditches. Methods A semi-dry and semi-wet ditch with O. hupensis snails was selected in the second branch field of Jiangbei Farm, Jiangling County, Hubei Province in May 2023, and divided into 4 experimental areas, named groups A1, A2, B1 and B2. Environmental cleaning was performed in groups A1 and B2, and was not conducted in groups A2 or B2. Then, 50% wettable powder of niclosamide ethanolamine salt was sprayed with drones at an effective dose of 2 g/m2 in groups A1 and A2, and 5% niclosamide ethanolamine salt granule was sprayed with drones at an effective dose of 2 g/m2 in groups B1 and B2. O. hupensis snails were surveyed using the systematic sampling method 1, 3, 5, 7, 14 days after spraying, and the natural mortality and corrected mortality of O. hupensis snails were calculated. Results The occurrence of frames with living snails, mean density of living snails and natural mortality of snails were 97.50% (117/120), 6.30 snails/0.1 m2 and 1.18% (9/765) in the test ditch before spraying, respectively. There were significant differences in the mortality of snails among four groups 1, 3, 5, 7 and 14 days after spraying niclosamide formulations with drones (χ2 = 17.230, 51.707, 65.184, 204.050 and 34.435, all P values < 0.01). The overall mortality rates of snails were 94.51% (1 051/1 112), 79.44% (908/1 143), 96.54% (977/1 012) and 88.55% (1 021/1 153) in groups A1, A2, B1 and B2 (χ2 = 207.773, P < 0.05), respectively. In addition, there was no significant difference in the overall snail mortality between groups A1 and B1 (P > 0.05), and the snail mortality in groups A1 and B1 were both statistically different from that in groups A2 and B2 (all P values < 0.05). Conclusion Both 50% wettlable powder of niclosamide ethanolamine salt and 5% niclosamide ethanolamine salt granule sprayed with drones are active against O. hupensis snails in ditches, and environmental cleaning may improve the molluscicidal effect.