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The world is suffering from the monkeypox epidemic, and China is facing a great risk of monkeypox importation. Understanding and mastering clinical manifestations, diagnosis and treatment of monkeypox is one of the important measures to respond to future crises. This review summarizes updated guidelines and relevant studies, and covers main clinical manifestations, diagnosis, treatment and follow-up of monkeypox patients, as well as management of special populations, aiming to provide references for clinicians and prevention workers.
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Monkeypox is a zoonotic disease caused by monkeypox virus, and human cases infected with the virus have been reported in more than 100 countries. To respond to the potential of case importation and consequent spread of the infection in the country, it is urgent for China to strengthen its comprehensive surveillance efforts consisting of case detection through country-entering check, symptom screening, and investigation among priority populations, and to implement comprehensive strategies to control the source of infection, interrupt the transmission and protect the people at risk.
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Neurosyphilis is one of the most serious clinical manifestations of syphilis. In recent years, more and more neurosyphilis cases have been reported in Chinese literature. However, the exact incidence of neurosyphilis is unknown, and there are still some key scientific problems urgent to be solved in clinical diagnosis and treatment, pathogenesis, and prevention and control of neurosyphilis. Therefore, in order to provide a reference for the prevention and treatment of neurosyphilis, this article proposes the CARE-NS strategy, including the following 6 aspects: Comprehensive management including multiple disciplinary treatment (C) , Alleviating neurological impairment and sequelae (A) , Risk factors and clinical epidemiology (R) , Etiology and pathogenesis (E) , New diagnostic indicators and strategies (N) , Social impact and cost-effectiveness analysis (S) .
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Objective:To investigate the phagocytosis of Treponema pallidum (Tp) by macrophages and the polarization direction of macrophages after Tp stimulation. Methods:Human THP-1 monocyte-derived M0 macrophages were stimulated with the Tp Nichols strain, and the phagocytosis of Tp by macrophages and changes in the intracellular structure of macrophages were observed by transmission electron microscopy and immunofluorescence staining. After 12-hour stimulation by Tp, Tp was removed, the M0 macrophages continued to be cultured for 24, 48, 72 hours and 6 days. Western blot analysis and immunofluorescence staining were performed to determine the expression of the M1 macrophage marker CD86 and M2 macrophage marker CD163, and enzyme-linked immunosorbent assay was conducted to detect levels of M1-type cytokines interleukin (IL) -12 p70, interferon (IFN) -γ, chemokine ligand 10 (CXCL10) , IL-6, tumor necrosis factor (TNF) -α and IL-1β, as well as the M2-type cytokine transforming growth factor (TGF) -β1 in the culture supernatant of macrophages. Dunnett- t test was used for multiple comparisons. Results:As transmission electron microscopy showed, after the stimulation by Tp, the macrophages extended pseudopodia and engulfed Tp, leading to swelling and obviously irregular hyperplasia of endoplasmic reticulum as well as enlargement of mitochondria. Moreover, after additional culture for 24, 48, 72 hours and 6 days, CD86 was highly expressed, but CD163 was lowly expressed in the Tp-treated macrophages; at 24 hours, the supernatant levels of IL-12 p70, IFN-γ, CXCL10, IL-6, TNF-α and IL-1β were significantly higher in the Tp-treated group than in the control group (all P<0.001) , but there was no significant difference in the TGF-β1 supernatant level between the 2 groups ( P>0.05) . Conclusions:After engulfment of Tp, the structures of endoplasmic reticulum and mitochondria in THP-1-derived macrophages markedly changed. Tp could induce the polarization of M0 macrophages into M1 macrophages, and phenotypic switch from M1 to M2 macrophage polarization was not observed within 6 days after Tp stimulation.
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Neurosyphilis is a serious clinical stage of syphilis caused by Treponema pallidum invading the nervous system, and risk and predictive factors of neurosyphilis are different between syphilis patients with and without HIV infection. The risk factors for neurosyphilis in HIV-negative patients with syphilis mainly include gender, age, clinical stage of syphilis, treatment, etc.; the predictive factors include serological titers, changes in some indicators of cerebrospinal fluid, neurological or ophthalmic symptoms. HIV viral load, CD4 + T cell counts and antiretroviral treatment are the main predictors and risk factors for neurosyphilis in HIV-positive patients with syphilis.
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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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With the development of genome sequencing technology,Treponema pallidum has been found to have different subspecies all around the world. It is widely recognized that Treponema pallidum could be subtyped by analyzing two or three target genes. Advances in molecular epidemiology of syphilis reveal that clinical characteristics of Treponema pallidum,such as virulence,serofast,drug resistance and the site of in-fection,are related to its subspecies. Specifically,14f/f may be more virulent;serofast may be more likely to happen in patients infected with Treponema pallidum of Tpr i genotype;A2058 and A2059 genes may be re-lated to resistance to macrolides. All these will be summarized in this review.
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Objective To evaluate the effects of Treponema pallidum (T.pallidum) on the expression of chemokine ligands (CXCL) in human brain microvascular endothelial cells (HBMECs).Methods HBMECs were randomly divided into 4 groups,which were treated with viable T.pallidum suspension (T.pallidum group),heat-inactivated T.pallidum suspension (inactivated T.pallidum group),200 μg/L lipopolysaccharide (LPS group) and cell culture medium (blank control group),respectively,for 6,12 and 24 hours.Fluorescence-based quantitative PCR and enzyme-linked immunosorbent assay (ELISA) were performed to determine the mRNA and protein expression of CXCL6,CXCL8 and CXCL10 in HBMECs in the above groups respectively.Transwell migration assay was conducted to evaluate the effects of T.pallidum-stimulated HBMECs on the chemotaxis of human promyelocytic HL-60 leukemia cells (HL-60 cells).Results At 6,12 and 24 hours,the T.pallidum group showed significantly higher mRNA expression of CXCL6,CXCL8 and CXCL10 in HBMECs compared with the blank control group and inactivated T.pallidum group (all P ＜ 0.05),while there were no significant differences between the blank control group and inactivated T.pallidum group (all P ＞ 0.05).Compared with the LPS group,the T.pallidum group showed significantly decreased mRNA expression of CXCL6 and CXCL8 (P ＜ 0.05),but similar mRNA expression of CXCL10(P ＞ 0.05)at 6,12 and 24 hours.At these time points,the levels of CXCL6 and CXCL8 in the culture supernatant of HBMECs were significantly higher in the T.pallidum group than in the blank control group and the inactivated T.pallidum group (all P ＜ 0.05),but no significant differences were observed between the blank control group and the inactivated T.pallidum group (both P ＞ 0.05).Moreover,there were no significant differences in the level of CXCL10 in the culture supernatant of HBMECs among the T.pallidum group,the inactivated T.pallidum group and the blank control group (all P ＞ 0.05).The number of migratory HL-60 cells in the lower Transwell chambers was significantly higher in the T.pallidum group than in the inactivated T.pallidum group and the blank control group (both P ＜ 0.05).Conclusion Viable T.pallidum can up-regulate the gene expression of CXCL6,CXCL8 and CXCL10 in HBMECs,promote the secretion of CXCL6 and CXCL8,and enhance the chemotactic effect of HBMECs on HL-60 cells,which may play a certain role in the occurrence of neurosyphilis.
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Objective To investigate characteristics of Treponema pallidum (Tp)-induced macrophage-derived exosomes and its effect on the proliferation of human umbilical vein endothelial cells (HUVEC).Methods Tp strains were collected from the testis of male rabbit,which were infected with Tp (Nichols strain).The human mononuclear macrophages (THP-1) were induced into macrophages by incubation with propylene glycol methyl ether acetate (PMA),and then the macrophages were divided into 2 groups:experimental group incubated with Tp for 12 hours followed by 48-hour normal culture,and control group receiving normal culture.After the treatment,exosome suspensions were collected,and exosomes were extracted by differential centrifugation and exoEasy Maxi Kit.Transmission electron microscopy and Western blot analyses were performed to identify the exosomes,and nanoparticle tracking analysis (NTA) was conducted to measure the diameters and concentrations of exosomes.In vitro cultured HUVECs were divided into 3 groups,which were cultured with the 10 μl of suspensions containing exosomes derived from Tp-stimulated macrophages at a concentration of 4.5 × 108/ml (experimental group),10 μl of suspensions containing exosomes derived from untreated macrophages at a concentration of 4.5 × 108/ml (control group),and 10 μl of exosome eluents (exosome eluent group),respectively.After the treatment,confocal laser scanning microscopy was performed to observe the phagocytosis of exosomes of HUVECs,and cell counting kit-8 to evaluate the proliferative activity of HUVECs.Results The exosomes were saucer-like microvesicles with diameters of 30-100 nm under the transmission electron microscope.Western blot analyses showed that membrane proteins CD63,CD9 and CD81 were abundantly expressed by exosomes.Under the same conditions,NTA revealed that there were no significant differences in the particle diameter (u =1.90,P ＞ 0.05) and concentration of exosomes (Z =-1.604,P =0.109) between the experimental group and the control group.After co-culture with HUVECs for 5 hours,confocal laser scanning microscopy showed scatteredly distributed exosomes with green fluorescence in the HUVECs in the experimental group and control group.After 12-hour co-culture with the exosome suspensions,the proliferative activity of HUVECs was significantly higher in the experimental group and the control group than in the exosome eluent group (both P ＜ 0.05).After 24-and 48-hour treatment with exosome suspensions,the proliferative activity of HUVECs in the control group was still significantly increased compared with that in the exosome eluent group,and peaked at 48 hours (all P ＜ 0.05).Moreover,there were no significant differences in the proliferative activity of HUVECs between the experimental group and exosome eluent group at 24 and 48 hours (both P ＞ 0.05).At 72 hours,no significant differences in the proliferative activity of HUVECs were observed among the experimental group,the control group and the exosome eluent group (all P ＞ 0.05).Conclusions The exosomes secreted by THP-1 cells-derived macrophages evidently increased the proliferative activity of HUVECs within 48 hours,which peaked at 48 hours.After the stimulation with Tp,the exosomes secreted by THP-1 cells-derived macrophages were similar to those without Tp stimulation in morphology,size and concentration,and only increased the proliferative activity of HUVECs within 12 hours.
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Objective To investigate the association of tumor necrosis factor-ot (TNF-α) gene promoter polymorphisms with generalized pustular psoriasis.Methods Totally,91 patients of Han nationality with generalized pustular psoriasis (generalized pustular psoriasis group) and 102 health checkup examinees (healthy control group) were enrolled into this study.PCR and direct sequencing were performed to analyze the-238,-308 and-857 polymorphic sites of the TNF-α promoter.Results The frequency of the A allele at TNF-α-238 site was significantly higher in the generalized pustular psoriasis group than in the healthy control group (P =0.003,OR =4.819,95％ CI:1.581-14.694),so was the frequency of GA/AA genotype (P =0.006,OR =4.455,95％ CI:1.410-14.077).However,no significant differences were observed in the frequencies of G/A alleles (P =0.794) and GG/GA/AA genotypes (P =0.786) at TNF-o-308 site,or in the frequencies of C/T alleles (P =0.474) and CC/CT/TT genotypes (P =0.453) at TNF-α-857 site,between the generalized pustular psoriasis group and healthy control group.Conclusion TNF-α-238G ＞ A polymorphisms may be associated with the occur-rence of generalized pustular psoriasis.
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Objective To investigate mechanisms underlying the regulation of the permeability of vascular endothelial cells by the Treponema pallidum membrane protein Tpp47.Methods Human umbilical vascular endothelial cell (HUVEC) monolayers were established as a model,and were directly cultured with the presence of recombinant Tpp47 protein (rTpp47-treated group),or boiled and inactivated rTpp47 (negative control group).Some HUVEC monolayers,which were pretreated with the RhoA/ROCK signal pathway inhibitor Y-27632 for 30 minutes and then cultured with the presence of rTpp47,served as the pretreatment group.After 1-and 4-hour additional culture,enzymelinked immunosorbent assay (ELISA) was performed to estimate the permeability of these cell monolayers to horseradish peroxidase (HRP).After 12 hours of culture,rhodamine-phalloidin was used to stain cytoskeletal proteins,and confocal laser scanning microscopy was performed to observe the arrangement of the cytoskeletal protein F-actin.Western-blot analysis was conducted to measure the expressions of RhoA in HUVECs treated with rTpp47 or inactivated rTpp47.Results The supernatant level of HRP (expressed as the absorbance value at 450 nm) was significantly higher in the rTpp47-treated group than in the negative control group (0.81 ± 0.10 vs.0.39 ± 0.09,P ＜ 0.05),but no significant difference was observed between the pretreatment group (0.51 ± 0.10) and rTpp47-treated group or negative control group (both P ＞ 0.05) after 1-hour culture.Similarly,the rTpp47-treated group showed significantly increased levels of HRP compared with the pretreatment group and negative control group (2.31-± 0.14 vs.1.21 ± 0.12 and 0.73 ± 0.12,both P ＜ 0.05),while there was no significant difference between the pretreatment group and negative control group after 4-hour culture.The expression of RhoA in HUVECs treated with rTpp47 was significantly higher than that in those treated with inactivated rTpp47.Confocal laser scanning microscopy showed that rTpp47 treatment led to the rearrangement of F-actin in HUVECs followed by the formation of stress fibers in cytoplasm,while Y-27632 could partly inhibit the rearrangement of F-actin.Conclusion The recombinant Treponema pallidum membrane protein Tpp47 can regulate the permeability of vascular endothelial cells through the RhoA/ROCK signal pathway.
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Objective To investigate the effects of broadband ultraviolet B (BB-UVB) on the proliferation of, tyrosinase activity and melanogenesis in melanocytes.Methods Melanocytes isolated from human foreskin were subjected to primary culture.Some cultured primary melanocytes were irradiated with BB-UVB at 10, 20, 30, 40, 50, 100, 200 and 300 mJ/cm2.Then, CCK-8 assay was performed to evaluate the proliferative activity of melanocytes, dopa oxidation assay to estimate the activity of tyrosinase, and sodium hydroxide (NaOH)-lysis method was used to determine melanin content.Real-time fluorescence-based quantitative PCR was conducted to measure the mRNA expressions of genes involved in non-canonical Wnt pathways in melanocytes after irradiation with BB-UVB at 30, 50 and 100 mJ/cm2.Western blot was carried out to determine the expressions of proteins involved in non-canonical Wnt pathways in melanocytes before and after irradiation with BB-UVB of 100 mJ/cm2.The melanocytes receiving no treatment served as the control group.Statistical analysis was carried out by one-way analysis of variance followed by least significant difference (LSD)-t test for multiple group comparisons and by the independent sample t test for two-group comparisons.Results After irradiation with BB-UVB at 10-300 mJ/cm2, the proliferative activity of melanocytes was gradually reduced compared with the control group (all P ＜ 0.05), and the survival rate of melanocytes was less than 50％ when the irradiation dose of BB-UVB was higher than 100 mJ/cm2.Furthermore, tyrosinase activity gradually increased in melanocytes after irradiation with BB-UVB at 10-100 mJ/cm2 compared with the control group, and the increase was statistically significant at the radiation dose of 100 mJ/cm2 (P ＜ 0.05).Compared with the control group, the WIF-1 mRNA expression level decreased, while c-Jun N-terminal kinase (JNK), microphthalmia-associated transcription factor (MITF), Ras-related C3 botulinum toxin substrate 1 (RAC 1) and tyrosinase (TYR) mRNA expression levels increased in melanocytes after irradiation with BB-UVB at 30, 50 and 100 mJ/cm2 (all P ＜ 0.05);the WNT5A mRNA expression significantly decreased in melanocytes irradiated with 30 and 50 mJ/cm2 BB-UVB, but increased in those irradiated with 100 mJ/cm2 BB-UVB (all P ＜ 0.05).The radiation with 100 mJ/cm2 BB-UVB significantly decreased the expression of WIF-1 protein, but enhanced the expressions of WNT5A, JNK, MITF, RAC1 and TYR proteins in melanocytes compared with the control group (all P ＜ 0.05).Conclusions BB-UVB can decelerate the proliferation of, elevate tyrosinase activity and melanin level in, melanocytes.The WIF-1 gene may inhibit melanogenesis, and the decrease in its expression may promote melanogenesis by activating the JNK/MITF/TYR pathway through the combined effects of proteins involved in non-canonical Wnt pathways.
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Objective To investigate the in vitro effects of Treponema pallidum membrane protein Tpp17 on the permeability of endothelial barrier for further investigation on the immunopathogenesis of syphi-lis.Methods A cellular model of in vitro monolayer was established by using human umbilical vein endo-thelial cells ( HUVECs) .Cell-ELISA and a TMB kit were respectively used to measure the expression of VE-cadherin and the flux of horseradish peroxidase ( HRP) by monolayer HUVECs after stimulation with the re-combinant Tpp17 (rTpp17) protein.THP-1 cells stained with Calcein AM were added to the top of HUVEC monolayer in Transwell culture.Then, the numbers of THP-1 cells in the upper wells and beneath the HUVEC monolayer were counted by using a fluorescence microscope.The rTpp17 protein-treated HUVECs were fixed in 4%buffered paraformaldehyde and stained with rhodamine-phalloidin for observing the distri-bution of F-actin under a confocal laser scanning microscope.Results Compared with the control group, the expression of VE-cadherin in HUVECs was decreased, while the permeability of HUVEC monolayer was increased upon the stimulation with rTpp17 protein (P<0.05).Moreover, rTpp17 protein-induced F-actin redistribution and increased transendothelial migration of THP-1 cells were observed in rTpp17 protein-trea-ted HUVECs as compared with those of the control group (P<0.05).Conclusion Treponema pallidum membrane protein Tpp17 could suppress the expression of VE-cadherin and enhance the redistribution of F-actin, resulting in an enhanced transendothelial migration of THP-1 cells and an increased permeability of HUVEC monolayer.The Tpp17 protein might play an important role in the immunopathogenesis of syphilis.
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Objective To observe the attachment of Treponema pallidum to human brain microvascular endothelial cells (HBMECs) in vitro.Methods Some primary cultured HBMECs were inoculated into in 24-well plates to be cocultured with the suspension of T.pallidum at a concentration of 1.6 × 107 treponemes/ml.After 0.5,2 and 4 hours of co-culture,scanning electron microscopy was conducted to observe the attachment of T.pallidum to HBMECs.Some HBMECs were cocultured with the presence of T.pallidum suspensions at different concentrations (4 × 106,8 × 106,1.6 × 107 treponemes/ml) for 2,4,6 and 16 hours,then,dark-field microscopy was performed to count the number of treponemes that attached to single HBMECs.Statistical analysis was carried out by using repeated-measures analysis of variance.Results As scanning electron microscopy showed,treponemes gathered at some regions on the surface of HBMECs when they attached to HBMECs.In addition,T.pallidum partly merged with the membrane of HBMECs at the site of attachment.After co-culture with T.pallidum suspensions,the number of treponemes that attached to single HBMECs was significantly different among different time points (F =387.72,P ＜ 0.001) and among different concentrations of T.pallidum suspensions (F =593.23,P ＜ 0.001),with an interaction effect between the concentration of T.pallidum suspensions and incubation period (F =98.74,P ＜ 0.001).Concretely speaking,the number of treponemes that attached to single HBMECs increased over time until 6 hours after the start of coculture,then showed a decreasing trend,and reached the nadir value at 16 hours.Conclusion T.pallidum can adhere to cultured HBMECs in vitro,likely by the merger of its end with the membrane of HBMECs at some regions.
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Objective To establish an in vitro model of mycobacterial granuloma.Methods Mononuclear cells were isolated from peripheral blood of healthy human subjects,and stimulated to differentiate into macrophages,which were then classified into four groups to be cocultured with Mycobacterium marinum,Mycobacterium tuberculosis,Bacillus Calmette-Guérin,and Mycobacterium leprae,respectively,for five days followed by incubation with peripheral blood mononuclear cells (PBMCs) from the corresponding donors to establish an in vitro model of mycobacterial granuloma.The macrophages cocultured with PBMCs or mycobacteria alone served as the control.Microscopy was performed to dynamically visualize the formation of granuloma in vitro,flow cytometry to detect the expressions of cell surface antigens at different stages,real-time quantitative PCR and enzyme-linked immunosorbent assay (ELISA) to determine the mRNA expressions of important cytokines and their protein levels in the supernatant of macrophages,respectively.Results After 7-9 days of coculture with mycobacteria and PBMCs,the macrophages aggregated to form granuloma-like clumps,and some cells fused to form multinuclear giant cells,along with the expressions of some surface antigens such as CD14,CD68 and CD86 on these macrophages.The mRNA expressions of some important cytokines,including tumor necrosis factor-a,interferon-γ interleukin (IL)-1 β and IL-10,were detectable in the macrophages cocultured with mycobacteria and PBMCs,and the secretion of these cytokines was confirmed by ELISA in the supernatant of these cells.Conclusions An in vitro model of mycobacterial granuloma is basically established,which may facilitate the investigation into the formation of granuloma caused by and immune response to mycobacterial infection.
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Objective To evaluate the effect of Treponema pallidum membrane protein Tpp47 on vascular endothelial cells.Methods Human umbilical vein endothelial cells (HUVECs) were classified into multiple groups to be cultured with various concentrations (50,100,200,400 and 800 μg/L) of the recombinant protein Tpp47 or lipopolysaccharide (LPS) for different durations (3,6,12,24 and 48 hours).Then,enzyme-linked immunosorbent assay (ELISA) was performed to determine the levels of intercellular cell adhesion molecule-1 (ICAM-1) and E-selectin in the culture supernatant of,fluorescence-based real-time quantitative PCR to quantify the mRNA expressions of ICAM-1 and E-selectin in,HUVECs.The 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT) assay was used to evaluate the proliferation activity of HUVECs treated with Tpp47 (400 μg/L) and LPS (200 μg/L) respectively for 24 hours.To estimate the effect on adhesion ability,some HUVECs were pretreated with Tpp47 (400 μg/L) and LPS (200 μg/L) respectively for 24 hours followed by coculture with THP-1 human monocytic leukaemia cells for 6 hours,then,the adhesion of HUVECs to THP-1 cells was visualized by fluorescence microscopy.The cells receiving no treatment served as the blank control.Results A significant increase was observed in the supernatant level (expressed as the absorbance value at 450 nm) of ICAM-1 for HUVECs treated with Tpp47 of 400 μg/L for 24 hours (1.28 ± 0.03 vs.0.90 ± 0.01,t =18.28,P ＜ 0.05) and that of E-selectin for HUVECs treated with Tpp47 of 400 μg/L for 12 hours (0.51 ± 0.01 vs.0.13 ± 0.03,t =18.19,P＜ 0.05) compared with untreated HUVECs.The adhesion rate to THP-1 cells was significantly higher in HUVECs pretreated with Tpp47 for 24 hours than in untreated HUVECs (56.1％ ± 1.9％ vs.16.3％ ± 2.1％,x2 =12.65,P ＜ 0.05).The cell proliferation rate was 19.5％ ± 1.7％ in HUVECs treated with Tpp47,significantly higher than that in untreated HUVECs (10.0％ ± 3.1％,x2 =3.92,P＜ 0.05),but lower than that in those treated with LPS (41.2％ ± 3.7％,x2 =10.42,P ＜ 0.05).Conclusions The recombinant membrane protein Tpp47 could enhance HUVECs to proliferate and adhere to monocytic THP-1 cells,suggesting a certain role of Tpp47 in the pathogenesis of syphilis.
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Objective To study and evaluate the effect of developing the preventive service in venereal clinic .Methods The questionnaire survey on the selected outpatients was performed by the trained medical personnel in the venereal disease clinic of 5 medical institutions in Chongqing .The intervention services were carried out by providing healthy advices and tailor-made preven-tive packs .The healthy knowledge publicity were posted in the waiting area of the venereal clinic .The questionnaire investigation in the re-visiting patients was performed again to understand the awareness of prevention venereal disease knowledge and the condom use .Results 96 .4% of outpatients in this study accepted the AIDS-related knowledge at the first visit .84 .1% of them acquired the knowledge by consulting their doctor ,60 .9% of the patients insisted on using condom in sexual activity after accepting the interven-tion knowledge ,which was much higher than 17 .4% before intervention .Conclusion The STD clinic attenders satisfy the interven-tional preventive service in the venereal clinic ,the improvement of the venereal disease related knowledge and the adverse behaviors is obvious .
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Objective To study the effects of Treponema pallidum membrane protein Tpp17 on ac-tivation of human umbilical vein endothelial cells (HUVECs) in vitro and to understand its role in the immu-nopathogenesis of syphilis .Methods HUVECs were co-cultured with recombinant protein Tpp 17.Then the expressions of TNF-α, MCP-1, ICAM-1 and E-selectin at mRNA and protein levels in supernatants were re-spectively detected by enzyme-linked immunosobent assay ( ELISA ) and fluorescent real-time quantitative PCR.The adhesive ability of THP-1 cells was observed by fluorescence microscopy after co-cultured pretrea-ted HUVECs with recombinant protein Tpp 17 with Calcein-AM labeled THP-1.Pretreated HUVECs were cultured in the lower chamber of Transwell with recombinant protein Tpp 17 and monocytic THP-1 cells were cultured in the upper chamber of Transwell .After that, the migration of monocytic THP-1 cells was evalua-ted by using fluorescence microscopy .Results Compared with the blank control group , the expression of TNF-α, ICAM-1, E-selectin, especially the MCP-1, were enhanced by recombinant protein Tpp 17 of Trepo-nema pallidum.Moreover, Tpp17 improved the adhesive ability and migration of monocytic THP-1 cells, es-pecially the latter.Conclusion Treponema pallidum membrane protein Tpp17 might play a certain role in the immunopathogenesis of syphilis by enhancing the expression of TNF-α, MCP-1, ICAM-1 and E-selectin, and by promoting the adherence ability and migration of monocytic THP-1 cells.
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ObjectiveTo clone and express the Tp0136 gene of Treponema pallidum,to purify the recombinant protein and to evaluate its immunocompetence.MethodsThe full-length Tp0136 gene was synthesized,then subcloned into the expression vector pET28a to construct a recombinant plasmid,pET28a-Tp0136,which was subsequently transfected into E.coli Rosetta for protein expression.The recombinant protein was purified with nickel-nitrilotriacetic acid(Ni-NTA) affinity chromatography,and identified by using sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE) and Western blot.New Zealand rabbits were immunized with the recombinant Tp0136 (rTp0136)protein,and anti-Tp0136 polyclonal antibodies in sera of the rabbits were examined by indirect enzyme linked immunosorbent assay(ELISA) with rTp0136 protein as the coating antigen.Also,positive sera were obtained from patients with syphilis and examined by Western blot to identify the immunoreactivity of the rTp0136 protein.ResultsThe E.coli expression vector pET28a-Tp0136 was constructed successfully,and rTp0136 protein was also successfully expressed with a molecular weight of about 50 kD and a purity above 95％.High titres of anti-rTp0136 antibodies were detected in sera of rabbits immunized with the rTp0136 protein,and Western blot showed that the rTp0136 could specifically react with the sera from syphilitic patients,which proved the high immunogenicity and immunoreactivity of the recombinant protein.ConclusionsThe full-length Tp0136 membrane protein is successfully expressed with a high immunocompetence,and Tp0136 membrane protein may play an important role in the pathogenic mechanisms of Treponema pallidum.
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ObjectiveTo understand the clinical and histopathologic diagnostic criteria for aneurysmal fibrous histiocytoma(AFH).MethodsThe clinical and histopathological features of 5 patients with AFH were retrospectively reviewed.ResultsThere were 3 males and 2 females in these patients.All the tumors clinically manifested as dark erythematous or brown nodules.Three cases had a recent history of rapid growth.The lesions were located on the limbs(n =3),or chest and lower mandible(n =2).Histopathological examination of skin biopsies showed typical features of dermatofibroma,accompanied by many irregular cleftlikeorcavernousblood-filledspaceswithnumeroushemosiderinpigmentsinallofthesecases.Immunohistochemically,the tumor cells were immunoreactive to vimentin and CD68 but negative for CD34 or CD31.Conclusions In view of a history of recent rapid growth,the presence of hemorrhagic pseudocysts and high vascularity,AFH should be differentiated from angiosarcoma and angiomatoid fibrous histiocytoma.