1.Compound Betamethasone enhances the expression of olfactory marker protein in olfactory mucosa of mice injured by influenza virus
Guilian WAN ; Daofeng NI ; Jing GUAN
Basic & Clinical Medicine 2010;30(5):530-533
Objective To investigate the effects of compound Betamethasone on the expression of olfactory marker protein(OMP)in murine olfactory mucosa injured by influenza virus.Methods An animal model was developed by intranasal application of influenza virus to mice.Compound Betamethasone was injected i.p.(3.5 mg/kg)on day 2 and day 4 after the insult.The expression of OMP was tested by immunohistochemistry and Western blot.Results The expression of OMP was significantly downregnlated in the olfactory mucosa of influenza virus control group 1 and influenza virus control group 2;the expression of OMP was significantly upregulated in the olfactory mucosa of post-infection compound Betamethasone group 1 and post-infection compound Betamethasone group 2.Conclusion Compound Betamethasone can enhance the expression of OMP in the olfactory mucosa injured by influenza virus.
2.Drug sensitivity of thirty-four reference slowly growing mycobacteria to first and second line antituberculous agents
Hui PANG ; Guilian LI ; Kanglin WAN ; Ping YU
Chinese Journal of Zoonoses 2015;(10):914-918
Slowly growing mycobacteria (SGM ) are distributed in the environment ,for example in soil and dirty water . SGM can cause human infections ,especially lung diseases .In this article ,first and second line antituberculous agents were ex‐amined in order to identify the optimum drugs for the treatment of SGM disorders .The fewest SGM in our study (4/34) were susceptible to isoniazid .Rifampicin (13/34) and ethambutol (14/34) were effective against similar numbers of strains .Ofloxa‐cin (23/34) ,kanamycin (26/34 ) , tobramycin (26/34 ) and streptomycin (27/34 ) were active against most of the tested strains .Ciprofloxacin (31/34) ,levofloxacin (31/34) ,amikacin (33/34) and capreomycin (33/34) showed an excellent range of activity .Moxifloxacin (34/34) showed the widest range of activity against the SGM species .Among the tested SGM spe‐cies ,M .simiae and M .af ricanum were resistant to the highest number of drugs .M .szulgai and M .duvalii were susceptible to all the first and second line antituberculous agents tested .Overall ,the second‐line antituberculous agents were good candi‐dates for the treatment of infection by SGM species and can be widely used in the therapy of SGM diseases .
3.C-reactive protein and autoimmune diseases
Yang HAN ; Tianpen CUI ; Guilian CHENG ; Binbin WAN
Chinese Journal of Laboratory Medicine 2014;(11):876-878
Impaired clearance of apoptotic cells is important in the pathogenesis of autoimmune disease.C-reactive protein (CRP) is an acute phase protein that plays a major role in the regulation of the autoimmune and inflammatory response .CRP has a role in the clearance of bacteria and dying and altered cells through binding to phosphocholine and might also have more complex immunomodulatory functions . CRP function as opsonins for pathogens and dying and apoptotic cells through activation of the complement pathway and through binding to Fcγreceptors , and is associated with the clearance of apoptotic cells and nuclear antigen , thus becoming a protective molecule against pathogenic autoimmune responses in general . Measurement of serum CRP level is in widespread clinical use as a sensitive marker of inflammation and autoimmune disease , particularly in relation to the use of the CRP-based disease activity score in the evaluation of rheumatoid disease.
4.Analysis on drug resistance-associated mutations of multi-drug resistant Mycobacterium tuberculosis based on whole-genome sequencing in China
Min GAO ; Tingting YANG ; Guilian LI ; Rong CHEN ; Haican LIU ; Qian GAO ; Kanglin WAN ; Shuidong FENG
Chinese Journal of Epidemiology 2020;41(5):770-775
Objective:To analyze the resistance mutational profiles of multi-drug resistant Mycobacterium tuberculosis in China and the correlation between major mutation types and genotypes based on the whole-genome sequencing data. Methods:Search and download of the genome-wide sequencing data of M. tuberculosis published in China by August 2019 on NCBI database were conducted. Mutation frequency of drug resistance-related gene loci based on whole-genome sequencing was used to predict the molecular susceptibility of strains, and the correlation between mutation types and genotypes was analyzed. Results:According to the results of molecular resistance and susceptibility profiles, 1 024 MDR strains were identified from 2 019 M. tuberculosis strains. The major mutation types of resistance-related genes to common drugs were katG S315T (73.2%, isoniazid), rpoB S450L (63.1%, rifampicin), rpsL K43R (70.0%, streptomycin), embB M306V (37.4%, ethambutol), pncA_promoter T (-11)C (7.9%, pyrazinamide), gyrA A90V (32.3%, fluoroquinolones), rrs A1401G (67.7%, second-line injection drugs), fabG1_promoter C (-15) T (87.0%, Ethionamide), folC I43T (30.4%, P-aminosalicylic acid). Among them, the frequencies of katG S315T, embB M306V, rpsL K43R, gyrA A90V in lineage 2 were significantly higher than those in lineage 4, and folC I43T was only found in lineage 2. The proportion of katG S315T was significantly higher in the ancient Beijing genotype compared to the modern genotype, in contrast, the proportion of rpsL K43R was significantly higher in modern Beijing genotype, the differences were significant (all P<0.05). Conclusions:The results showed the main mutation types of resistance-related genes of MDR strains to many commonly used anti-tuberculosis drugs in China based on whole-genome sequencing, providing a basis for the development of sensitive and specific rapid molecular detection methods. At the same time, it was also found that the major mutation types of MDR-related genes were related to the genotype of the strains.
5.Preliminary evaluation of immunogenicity and protective effect of multicomponent recombinant protein vaccine EPRHP014 against tuberculosis
Bin CAO ; Xueting FAN ; Ruihuan WANG ; Xiuli LUAN ; Chengyu QIAN ; Jinjie YU ; Haican LIU ; Machao LI ; Guilian LI ; Xiuqin ZHAO ; Xiuqin YUAN ; Kanglin WAN
Chinese Journal of Epidemiology 2023;44(10):1653-1660
Objective:To evaluate the immunogenicity and protective effect of a multicomponent recombinant protein vaccine EPRHP014 constructed independently and provide a scientific basis for developing new tuberculosis (TB) vaccine and effective prevention and control of TB.Methods:Three full-length Mycobacterium ( M.) tuberculosis protein antigens (EsxH, Rv2628, and HspX) and two epitope-predicted and optimized epitope-dominant protein antigens (nPPE18 and nPstS1) were selected, from which five protein antigens were used to construct a protein antigen composition EPRHP014, including a fusion expression multi-component protein antigen (EPRHP014f) and a multi-component mixed protein antigen (EPRHP014m) formed with the five single protein using clone, purification, and purification respectively. Multicomponent protein vaccines EPRHP014f and EPRHP014m were prepared with aluminum adjuvant, and the BCG vaccine was used as a control. ELISA detected the titer of serum-specific antibodies, the secretion of various cytokines was detected by ELISpot and Luminex, and immune protection was observed by the M.tuberculosis growth inhibition test in vitro. The results were statistically analyzed by t-test or rank sum test, and P<0.05 was considered a statistically significant difference. Results:Mice Immunized with EPRHP014m and EPRHP014f could produce highly effective IgG antibodies and their subtypes IgG1 and IgG2a, and the antibody titers were similar to those of mice immunized with BCG, with no statistical significance ( P>0.05). The number of spot-forming cells (SFC) secreting IFN-γ and IL-4 induced by EPRHP014f group was significantly higher than those by EPRHP014m group and BCG group ( P<0.05), but there was no significant difference in the number of SFC for IFN-γ and IL-4 induced between EPRHP014m group and BCG group ( P>0.05). The secretion levels of GM-CSF and IL-12p70 induced by the EPRHP014m group were higher than those of the BCG group ( P<0.05), but there was no significant difference in the levels of IL-6 and IL-10 induced between EPRHP014m group and BCG group ( P>0.05). There was no significant difference in the secretions of IL-6, IL-10, IL-12, and GM-CSF between the EPRHP014f and BCG groups ( P>0.05). EPRHP014m group, EPRHP014f group, and BCG group had obvious antibacterial effects in vitro, and the difference was insignificant ( P>0.05). Conclusion:Both EPRHP014f and EPRHP014m can induce strong humoral and cellular immune responses in mice after immunization, and have a strong ability to inhibit the growth of M. tuberculosis in vitro, indicating that the antigen composition EPRHP014 has good potential in the development and application of TB vaccine.
6.Preliminary immunological evaluation of Mycobacterium tuberculosis multicomponent protein vaccine candidates EPDPA015f and EPDPA015m
Ruihuan WANG ; Xueting FAN ; Chengyu QIAN ; Bin CAO ; Jinjie YU ; Machao LI ; Guilian LI ; Xiuqin ZHAO ; Xiuli LUAN ; Haican LIU ; Kanglin WAN
Chinese Journal of Microbiology and Immunology 2023;43(4):294-303
Objective:To preliminarily evaluate the immunogenicity and efficacy of two novel tuberculosis vaccine candidates (a fusion multicomponent protein EPDPA015f and a mixed multicomponent protein EPDPA015m) and to provide a new antigen combination for the development of tuberculosis vaccines.Methods:Recombinant plasmids for the expression of EPDPA015f and EPDPA015m proteins were constructed. Six-week-old BALB/c mice were immunized with EPDPA015f or EPDPA015m in combination with aluminium adjuvant (50 μg/mouse) for three times with an interval of 10 d. The mice were sacrificed 10 d after the last immunization to collect blood and spleen samples. Serum antibody titers and cytokine levels were measured by ELISA, Luminex technique and enzyme-linked immunospot assay (ELISPOT). Mycobacterial growth inhibition assay (MGIA) was used to detect the ability of mouse splenocytes to inhibit the growth of Mtb in vitro. One-way analysis of variance and t-test were used for statistical analysis. Results:Both EPDPA015f and EPDPA015m could induce the production of various cytokines and IgG antibodies at a high level. The levels of cytokines related to Th1 (IL-2, TNF-α, IFN-γ), Th2 (IL-4, IL-6, IL-10) and Th17 (IL-17) as well as other proinflammatory cytokines (GM-CSF, IL-12) were higher in the EPDPA015f group than in the adjuvant group ( P<0.05). The titer of IgG antibody induced by EPDPA015f was as high as 1∶4×10 6. The results of MGIA showed that the numbers of Mtb (lgCFU) in the PBS, adjuvant, EPDPA015f and EPDPA015m groups were 3.46±0.11, 3.51±0.06, 2.98±0.09 and 3.19±0.08, respectively. The number of colonies in the EPDPA015f group was the least as compared with that in the other three groups ( P<0.001, P<0.001, P<0.01). Conclusions:The vaccine candidate EPDPA015f could elicit more comprehensive and high-level cellular and humoral immune responses, and exhibited superior in vitro inhibitory activity against the growth of Mtb. EPDPA015f had the potential to be used as a preventive vaccine or a booster vaccine