1.Susceptibility of Mycoplasma Hominis to Several Antimicrobials and Chinese Medicinal Herbs
Yamin CHE ; Shuhe MAO ; Wenling JIAO
Chinese Journal of Dermatology 1994;0(06):-
Objective To study the susceptibility of Mycoplasma hominis(Mh) to several antimicrobials and Chinese medicinal herbs. Methods The minimal inhibitory concentrations(MIC) of Mh to 11 antimicrobials and 19 herbs were detected by the method of liquid microdilution. Results The sensitivity of Mh to those antimicrobials from high to low, was as follows:lincomycin, tetracycline, minocycline, levofloxacin, ciprofloxacin, ofloxacin, rifampin, spiramycin, azithromycin, erythromycin ethyl succinate and erythromycin. Among 19 herbs,Mh was sensitive to Radix Isatidis, Radix Angelicae Dahuricae, Cortex Phellodendri, Rheam Officinale, Fructus Kochiae and Herba Houttuyniae. Conclusions The susceptibility of local isolates of Mh to common antimicrobials is found out. Six Mh sensitive Chinese medicinal herbs are screened. The study provides evidences of treating Mh infection by combination of antimicrobials with Chinese medicinal herbs.
2.Construction of a novel bivalent DNA vaccine against SARS-CoV-2 and influenza A virus H3N2 and evaluation of its immunogenicity in mice
Chengcheng ZHAI ; Di HAN ; Yao DENG ; Jiao REN ; Wen WANG ; Donghong WANG ; Wenling WANG ; Long GAO ; Wenjie TAN
Chinese Journal of Microbiology and Immunology 2022;42(9):683-690
Objective:To construct a bivalent DNA vaccine against SARS-CoV-2 and influenza A virus H3N2 and to evaluate its immunogenicity in mice.Methods:The coding sequences for spike 1 (S1) protein of SARS-CoV-2 Beta variant and hemagglutinin (HA) of influenza A virus Cambodia (H3N2) strain were codon-optimized and synthesized. The two coding genes were ligated by the self-cleaving 2A peptide using over-lapping PCR to construct S1-2A-HA fragment, which was inserted into pVRC vector to construct the bivalent DNA vaccine, named as pVRC-S1-2A-HA. Indirect immunofluorescence assay (IFA) and Western blot were performed to detect the expression of S1 and HA proteins. BALB/c mice were immunized with pVRC-S1-2A-HA by intramuscular injection and electroporation. The humoral immune responses induced in mice were detected by indirect ELISA, pseudovirus neutralization assay and hemagglutination inhibition assay. Cellular immune responses were detected by IFN-γ ELISPOT, intracellular cytokine staining (ICS) and cytometric bead array (CBA).Results:The bivalent DNA vaccine pVRC-S1-2A-HA could express S1 and HA proteins in vitro. Specific cellular immune responses against S1 protein and specific IgG antibody against HA protein were significantly induced in mice with single-dose immunization. The antigen-specific immunity was significantly enhanced after booster immunization. The geometric mean titer (GMT) of specific IgG antibody increased to 3 251 for S1 protein and 45 407 for HA protein after two-dose immunization. Moreover, the S1-specific T cells increased to 1 238 SFC/10 6 cells. ICS results indicated that the booster vaccination induced CD4 + T and CD8 + T cells to produce IL-2, IFN-γ and TNF-α in mice. The secretion of various cytokines including IL-2, IL- 4, IL-6, IL-10 and IFN-γ in mouse splenocytes was induced after single-dose immunization. Conclusions:A bivalent DNA vaccine against SARS-CoV-2 and influenza A virus H3N2 was constructed and could induce S1- and HA-specific humoral and cellular immune responses in mice, suggesting the great potential of it for further development and application.
3.Normalization and parallel comparison of the detection results of antibodies against SARS-CoV-2
Wenling WANG ; Huijuan WANG ; Baoying HUANG ; Yao DENG ; Li ZHAO ; Fei YE ; Wen WANG ; Jiao REN ; Wenjie TAN
Chinese Journal of Microbiology and Immunology 2022;42(10):761-768
Objective:To improve the consistency of test results through reducing inter-laboratory variation in SARS-CoV-2 antibody detection with WHO SARS-CoV-2 antibody candidate international standard (IS, sample G) and antibody reference panel (samples E, F, H, I, J).Methods:Ten WHO samples (A-J) including the candidate IS and reference panel were evaluated using different methods, such as microneutralization tests based on live SARS-CoV-2, pseudovirus neutralization assay and commercial ELISA kits. The test results were compared using statistical analysis.Results:Using IS (sample G) as a reference, the relative concentrations of other samples could be determined with less variation. ELISA and pseudovirus neutralization assay had consistent results with those obtained with the microneutralization test based on SARS-COV-2 strain HB02. Weakly positive samples could be detected only by a certain kit.Conclusions:The availability of an IS for antibodies would facilitate the standardization of SARS-CoV-2 antibody detection methods. The reference panel fitted all the assays based on the SARS-CoV-2 prototype Wuhan strain. Pseudovirus neutralization assay and ELISA could be used as alternatives to live SARS-CoV-2-based neutralization test to some extent.