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Neurosyphilis is an infection of the central nervous system caused by Treponema pallidum with complex and atypical clinical manifestations. As no gold standard has been established, the diagnosis of neurosyphilis mainly relies on a comprehensive analysis of epidemiological history, clinical manifestations and laboratory examinations. This review discussed the advances in laboratory examinations of neurosyphilis in order to provide reference for clinical diagnosis of neurosyphilis.
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Objective To certify that Chlamydia can spread from the genital tract to the gastrointestinal tract for long-lasting colonization.Methods Totally,120 female C57BL/6J mice aged 5-6 weeks were divided into 4 experimental groups to be inoculated with purified Chlamydia muridarum (C.muridarum) elementary bodies in the vagina (n =35),gastric area (n =30),anus and rectum (n =30),retro-orbital venous plexus (n =5) respectively.Moreover,corresponding negative groups inoculated with sucrose phosphate glutamate buffer (n =5) were set up for each experimental group.On days 3,7,and every 7 days,vaginal and rectal discharges were collected with swabs from the mice,and the number of live C muridarum orgnisms in exfoliated cells infected with C muridarum in the swabs was determined.Indirect immunofluorescence assay and quantitative PCR (qPCR) were performed to determine the number of live chlamydial organisms and the copy number of chlamydial genomes in the mouse genital tract (vagina,uterus,oviduct and ovary),gastrointestinal tract (stomach,small intestine,cecum,colon,rectum)and parenteral tissues (heart,liver,spleen,lung,kidney) on days 7,14,28,56 and 105 after the inoculation.The number of live chlamydial organisms and copy number of chlamydial genomes were transformed logarithmically with a base of 10.The degree of hydrosalpinx and inflammation in the genital tract,and histopathological changes of the gastrointestinal tract were observed.The infectivity and virulence of C.muridarum in the genital tract and gastrointestinal tract were evaluated in the intragastric inoculation group and intra-anal and intrarectal inoculation group on days 28 and 56 after the inoculation.Blood samples were obtained from the mouse caudal vein in the retro-orbital venous plexus inoculation group on days 3,5,7,10 and 14 after the inoculation,the number of live chlamydial organisms and the copy number of chlamydial genomes in the blood samples were determined,and chlamydial infectivity in the genital tract and gastrointestinal tract was evaluated on day 56.Results On day 7 after the inoculation in the vagina,both C.muridarum live organisms and genomes were detected in the genital tract,gastrointestinal tract and parenteral tissues of all the mice.The largest common logarithm of the number of C.muridarum inclusion forming units (IFU) was observed in the vagina (6.26 ± 0.56),with the common logarithm of the copy number of chlamydial genomes in the vagina being 7.30 ± 0.23,and the common logarithms of the number of Chlamydia IFU and genomic copy were 2.60 ± 1.95 and 4.87 ± 0.09 respectively in the rectum.On day 28,no live Chlamydia was detected in the heart,lung or other parenteral tissues,while live Chlamydia could be found in the genital tract and gastrointestinal tract.The common logarithms of the number of Chlamydia IFU and genomic copy were 3.47 ± 1.06 and 5.80 ± 1.49 respectively in the vagina,and 4.00 ±0.35 and 5.14 ± 0.81 respectively in the rectum.On day 56,live Chlamydia could only be detected in the gastrointestinal tract.On day 105,live Chlamydia and its genomes could be still detected in the gastrointestinal tract,and the common logarithms of the number of Chlamydia IFU and genomic copy could be up to 2.60 ± 0.65 and 4.29 ± 0.57 respectively in the rectum.On days 28 and 56 after the inoculation,both live Chlamydia and its genomes could be detected in the gastrointestinal tract of all the mice in the intragastric inoculation group and intra-anal and intrarectal inoculation group.Chlamydia could survive in the blood for about 14 days in the retro-orbital venous plexus inoculation group,and live Chlamydia was detected in anal-rectal swabs in all the mice on day 14.On day 56 after the intravaginal inoculation with C.muridarum,severe hydrosalpinx,chronic inflammation and oviduct dilation occurred in the genital tract of 5 mice,but there was no obvious infiltration of inflammatory cells in the gastrointestinal tract,and inflammatory pathological changes were not observed in the gastrointestinal tract of mice after inoculation with Chlamydia through other routes either.Conclusion The infection with Chlamydia in the genital tract can lead to systemic dissemination,and Chlamydia can be spread to the gastrointestinal tract,and colonize and survive in the gastrointestinal tract for a long time.
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Objective To construct active phages against Chlamydia trachomatis,and to evaluate its effect on Chlamydia trachomatis.Methods The M13 phage was recombined with the IN5 sequences encoding the capsid protein VP1 of chlamydiophage phiCPG1,and then the recombinant M13-IN5 phage was obtained.PCR amplification,enzyme digestion and sequencing were performed to verify whether the target fragment was inserted into the phage successfully.The viability of the phage was evaluated by plaque formation assay.Cell counting kit-8 (CCK8) assay was conducted to evaluate the effect of M13 phage and recombinant M13-IN5 phage at the titer of 1011 plaque-forming units (PFU)/ml on the proliferation of Hela cells,and Hela cells uninfected with chlamydia served as the blank control group.Western blot analysis was performed to determine the expression of the IN5 loop protein in the recombinant M13-IN5 phage,M13 phage and Escherichia coli ER2738 at exponential growth phase.Cultured standard Chlamydia trachomatis serovar E strain was treated with M13 phage and recombinant M13-IN5 phage at the titer of 1011 PFU/ml separately,and chlamydia control group without the treatment with phages was set up.After 36-hour infection,confocal microscopy was performed to detect the location of the M13 phage and the recombinant M13-IN5 phage.Moreover,iodine staining was conducted to count inclusion bodies at 36,48,60 and 72 hours separately after infection.Statistical analysis was carried out by a two-sample t-test for comparisons between two groups,one-way analysis of variance (ANOVA) for intergroup comparison,and Bonferroni test for multiple comparisons.Results The bioactive recombinant M13 phage containing the IN5 loop gene was constructed successfully,and Western blot analysis confirmed that the recombinant phage expressed IN5 loop/p Ⅲ fusion protein with a high titer of 3.05 × 1011 PFU/ml.As CCK8 assay showed,there was no significant difference in proliferation of Hela cells among the blank control group,M 13 phage group and recombinant M13-IN5 phage group (A450 values:3.63 ± 0.01,3.55 ± 0.02,3.70 ± 0.01,respectively,F =12.0,P ＞ 0.05).Confocal microscopy showed overlap between the phage fluorescence and chlamydial inclusion body fluorescence.The M13-IN5 phage group and M13 phage group both showed significantly decreased number of inclusion bodies compared with the control group (both P ＜ 0.05) at 36 and 72 hours after chlamydial infection,and the number of inclusion bodies was significantly lower in the M 13-IN5 phage group than in the M13 phage group (P ＞ 0.05).After 48,and 60 hours of chlamydial infection,the number of inclusion bodies did not differ among the M13 phage group,M13-IN5 phage group and control group (both P ＞ 0.05).Conclusions The recombinant M13-IN5 phage was bioactive and could successfully express the IN5 loop protein.In the in vitro experiments,the recombinant phage could enter into chlamydia inclusion bodies,and markedly inhibited the infection of Chlamydia trachomatis.