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A 46-year-old male patient developed scatterred reddish-brown plaques and nodules on the back 6 years prior to the presentation. Then, the lesions gradually spread to the axillary fossa and protothorax, and became indurated with slight itching in winter. Laboratory examination revealed hypergammaglobulinemia. Computed tomography(CT)scan showed multiple nodular or patchy shadows in both lungs, lymphadenectasis in axillary, mediastinal and inguinal regions, and spleen enlargement. Histopathological examination of skin lesions showed granulomatous infiltrates with plenty of lymphocytes, histiocytes and mature plasma cells in the middle and lower dermis with the presence of lymphoid follicle-like structures, but no cell atypia was observed. Immunohistochemical study showed positive staining for CD38, CD138, CD79a, κ and λ light chains. According to clinical manifestations and laboratory examination results, the patient was diagnosed with cutaneous and systemic plasmacytosis.
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Objective To compare the potentiality of opa typing versus Neisseria gonorrhoeae multi-antigen sequence typing (Ng-MAST) in discrimination of N. gonorrhoeae isolates, and to investigate the consistency of genotypes of N. gonorrhoeae between patients and their sex partners. Methods N. gonor-rhoeae was isolated from patients and their sex partners who attended sexually transmitted disease clinics of the Affiliated Union Hospital of Tongji Medical College, Huazhong University of Science and Technology from February 2006 to August 2007. All isolates were cultured on Thayer-Martin agar and GC-based choco-late agar, and identified by Gram stain and oxidase reaction. Totally, twenty-four strains of N. gonorrhoeae were isolated form 12 pairs of male-female sex partners. Genomic DNA was isolated from the strains, and subjected to PCR amplification of opa gene as well as pot and tbpB genes, opa typing and Ng-MAST typing were utilized to differentiate the 24 strains respectively. Results The 24 isolates were classified into 10 sequence types (ST) by Ng-MAST typing, and 12 opa types (OT) by opa typing. A novel genotype identi-fied by Ng-MAST (217-86% homologisation 178), was unique in China. Conclusions Both ST and OT of N. gonorrhoeae strains are identical between patients and their corresponding partners except for one pair of partners, 45/46, suggesting that gonorrhea is transmitted to each other by contiguity between sex partners. Opa typing is likely to be more potential than Ng-MAST in genotyping of N.gonorrhoeae strains.
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In order to investigate the role of MMP-9 and TIMP-1 in the pathogenesis of systemic sclerosis, the expression of MMP-9 and TIMP-1 was immunohistochemically detected in skin lesions of the patients with diffuse cutaneous systemic sclerosis, skin lesions of the patients with limited cutaneous systemic sclerosis, and skin tissues of normal subjects. The results showed that the expression of MMP-9 in lesions of diffuse cutaneous systemic sclerosis was significantly lower than that of normal skins (P<0.05). However, no significant difference in the level of MMP-9 in the limited cutaneous systemic sclerosis and normal skin was found. Meanwhile, the expression of TIMP-1 in lesions of diffuse cutaneous systemic sclerosis and limited cutaneous systemic sclerosis were significantly higher than that of normal skins (both P<0.05). It was suggested that the expression of MMP-9 and TIMP-1 might play an important role in the development of systemic sclerosis.
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In order to investigate the role of TNF-α and ICAM-1 in the pathogenesis of lichen planus, immunohistechemistty was used to detect the expression of TNF-α and ICAM-1 in skin le- sions of the patients with lichen planus and skin tissues of normal subjects. The results showed that positive rates of TNF-α and ICAM-1 expressions in lichen planus were significantly higher than those in normal skins (both P<0.05). Meanwhile, there was a obvious correlation between the in- crease of TNF-α and that of ICAM-1 in lichen planus. The expression of TNF-α and ICAM-1 might play an important role in the development of lichen planus.
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In order to study the expression of intedeukin-22 (IL-22) and S100A7, A8, A9 mRNA in the skin lesions of patients with psoriasis vulgaris and their relationship, the biopsies were taken from skin lesions in 35 patients with psoriasis vulgaris and the skin of 16 normal controls, and the expres- sion levels of IL-22 and S 100A7, A8 and A9 mRNA were detected by semi-quantitative RT-PCR. The results showed that (1) IL-22 and SI00A8, A9 mRNA were positively expressed in the psoriatic skin lesions but negatively expressed in the normal controls; The expression level of S100A7 was (1.133±0.040) in the psoriatic skin lesions, significantly higher than that in the normal controls (0.744±0.037, P<0.01). (2) There were significantly positive correlations between the expression of IL-22/S100A7 mRNA, IL-22/S100A8 mRNA, IL-22/S100A9 mRNA in the psoriasis vulgaris (r1=0.543, r2=0.774, r3=0.621, P<0.01). It was concluded that IL-22 and S100A7, A8, A9 might play important roles in the occurrence and progression of psoriasis.
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In order to investigate the role of Th17 cytokines in the pathogenesis of psoriasis, reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect the expression of IL-17, IL-23 (p19/p40), and IL-6 in skin lesions and non-lesions of the patients with psoriasis and skin tissues of normal subjects. The results showed that the mRNA expression levels of IL-17, IL-23p19, IL-23p40 and IL-6 in psoriasis leision were significantly higher than those of non-leisions (1.231±0.843 vs 1.003±0.044, 1.166±0.142 vs 0.765±0.133, 1.125±0.104 vs 0.730±0.103, 1.186±0.222 vs 0.976±0.122, respectively, all P＜0.05). Meanwhile, The expression levels of IL-17 mRNA,IL-23p19 mRNA, IL-23p40 mRNA and IL-6 mRNA were higher in non-leisions than those in normal skin tissues (1.003±0.044 vs 0.620±0.104, 0.765±0.133 vs 0.584±0.078, 0.730±0.103 vs 0.000±0.000, 0.976±0.122 vs 0.656±0.121, respectively, all P＜0.05). The overexpression of Th17 cytokines in the skin lesions of patients with psoriasis may indicate that Th17 cytokines play a very important role in the immunopathogenesis of psoriasis.
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To study the expression of p63 and cyclooxygenase-2 (cox-2) in skin tumors and evaluate the correlation between p63 and cox-2, the expressions of cox-2 and p63 were measured by streptavidin-peroxidase complex immunohistochemical technique in 17 cases of skin squamous cell carcinoma (SCC), 19 cases of Bowen's disease(Bowen), 11 cases of actinic keratosis(AK), 12 cases of seborreic keratosis(SK) and 13 specimens of normal skin. Our results showed that the expression of p63 in skin squamous cell carcinoma, Bowen's disease and actinic keratosis were significantly higher than that in seborreic keratosis, while the expression of p63 in seborreic keratosis was significantly higher than that in normal skin. The expression of cox-2 in skin squamous cell carcinoma,Bowen's disease and actinic keratosis were significantly higher than that in seborreic keratosis, while no statistical difference was noted in the expression of cox-2 between seborreic keratosis and normal skin. Cox-2 expression was positively correlated with the high p63 expression in malignant skin tumors. The increased expression of cox-2 and p63 may play an important role in the development of skin tumors and work synergetically in malignant skin tumors.
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To investigate the role of toll-like receptor 9 (TLR9) in the pathogenesis of lichen planus,the expressions of TLR9 and its mRNA in the lesional skin of lichen planus were detected by immunohistochemical technique (SP) and RT-PCR. As control, normal skin of healthy volunteers was also tested. The immunohistochemical study showed that the expression of TLR9 in the lesional skin of lichen planus was significantly higher than that in the normal controls. The results of RT-PCR showed that both skin lesions and normal controls had TLR9 expression. In skin lesions, the expression level of TLR9 mRNA was 1.6075±0.0930, which was significantly higher than that in normal controls (P＜0.001). These findings indicated that up-regulated expression of TLR9 and its mRNA might be involved in the pathogenesis of lichen planus.
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In order to investigate the role of Th17 cytokines in the pathogenesis of psoriasis, reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect the expression of IL-17, IL-23 (p19/p40), and IL-6 in skin lesions and non-lesions of the patients with psoriasis and skin tissues of normal subjects. The results showed that the mRNA expression levels of IL-17, IL-23p19, IL-23p40 and IL-6 in psoriasis lesion were significantly higher than those of non-lesions (1.231 +/- 0.843 vs 1.003 +/- 0.044, 1.166 +/- 0.142 vs 0.765 +/- 0.133, 1.125 +/- 0.104 vs 0.730 +/- 0.103, 1.186 +/- 0.222 vs 0.976 +/- 0.122, respectively, all P < 0.05). Meanwhile, The expression levels of IL-17 mRNA, IL-23p19 mRNA, IL-23p40 mRNA and IL-6 mRNA were higher in non-lesions than those in normal skin tissues (1.003 +/- 0.044 vs 0.620 +/- 0.104, 0.765 +/- 0.133 vs 0.584 +/- 0.078, 0.730 +/- 0.103 vs 0.000 +/- 0.000, 0.976 +/- 0.122 vs 0.656 +/- 0.121, respectively, all P < 0.05). The overexpression of Th17 cytokines in the skin lesions of patients with psoriasis may indicate that Th17 cytokines play a very important role in the immunopathogenesis of psoriasis.
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To investigate the role of toll-like receptor 9 (TLR9) in the pathogenesis of lichen planus, the expressions of TLR9 and its mRNA in the lesional skin of lichen planus were detected by immunohistochemical technique (SP) and RT-PCR. As control, normal skin of healthy volunteers was also tested. The immunohistochemical study showed that the expression of TLR9 in the lesional skin of lichen planus was significantly higher than that in the normal controls. The results of RT-PCR showed that both skin lesions and normal controls had TLR9 expression. In skin lesions, the expression level of TLR9 mRNA was 1.6075+/-0.0930, which was significantly higher than that in normal controls (P<0.001). These findings indicated that up-regulated expression of TLR9 and its mRNA might be involved in the pathogenesis of lichen planus.
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To study the expression of p63 and cyclooxygenase-2 (cox-2) in skin tumors and evaluate the correlation between p63 and cox-2, the expressions of cox-2 and p63 were measured by streptavidin-peroxidase complex immunohistochemical technique in 17 cases of skin squamous cell carcinoma (SCC), 19 cases of Bowen's disease(Bowen), 11 cases of actinic keratosis(AK), 12 cases of seborreic keratosis(SK) and 13 specimens of normal skin. Our results showed that the expression of p63 in skin squamous cell carcinoma, Bowen's disease and actinic keratosis were significantly higher than that in seborreic keratosis, while the expression of p63 in seborreic keratosis was significantly higher than that in normal skin. The expression of cox-2 in skin squamous cell carcinoma, Bowen's disease and actinic keratosis were significantly higher than that in seborreic keratosis, while no statistical difference was noted in the expression of cox-2 between seborreic keratosis and normal skin. Cox-2 expression was positively correlated with the high p63 expression in malignant skin tumors. The increased expression of cox-2 and p63 may play an important role in the development of skin tumors and work synergetically in malignant skin tumors.
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In order to investigate the mRNA expression and function of interleukin-23 (p19/p40) and interleukin-12 (p35/p40) in the psoriatic lesion, no-lesion and normal human skin, reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect the expression of IL-23 (p19/p40) and IL-12 (p35/p40). The results showed that the expression of IL-23p19 mRNA and p40 (IL-12/IL-23)mRNA were higher in psoriatic lesion than those of non-lesional skin and normal skin. The levels of IL-23p19 mRNA and p40 (IL-12/IL-23) mRNA were higher in psoriatic non-lesional skin than normal skin. However, no significant difference was found in the level of IL-12p35 mRNA among the psoriatic lesional skin, non-lesional skin and normal skin. It was suggested that IL-23 might be more important in the pathogenesis of psoriasis than IL-12.
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To investigate the expression of plasmacytoid dendritic cells (pDCs), interferon regulatory factor-7 (IRF-7) and interferon alpha (IFN- α ) mRNA in skin lesions of patients with psoriasis vulgaris, the expressions of plasmacytoid dendritic cells, IRF-7, IFN-α mRNA in the lesional skin of psoriasis vulgaris were detected by immunohistochemical technique (SP) and RT-PCR. Normal skin of healthy volunteers, serving as control, was also tested. The immunohistochemical study showed that the expression of pDCs in the psoriatic lesions was significantly higher than that in the normal controls. RT-PCR showed that the mRNA expression of IRF-7 was much higher than that in normal controls, but no difference in the expression of IFN-α mRNA was found between two groups. Our findings indicate that up-regulated expression of pDCs, IRF-7mRNA might be involved in the pathogenesis of psoriasis.
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To investigate the expression of plasmacytoid dendritic cells (pDCs), interferon regulatory factor-7 (IRF-7) and interferon alpha (IFN- alpha) mRNA in skin lesions of patients with psoriasis vulgaris, the expressions of plasmacytoid dendritic cells, IRF-7, IFN-alpha mRNA in the lesional skin of psoriasis vulgaris were detected by immunohistochemical technique (SP) and RT-PCR. Normal skin of healthy volunteers, serving as control, was also tested. The immunohistochemical study showed that the expression of pDCs in the psoriatic lesions was significantly higher than that in the normal controls. RT-PCR showed that the mRNA expression of IRF-7 was much higher than that in normal controls, but no difference in the expression of IFN-alpha mRNA was found between two groups. Our findings indicate that up-regulated expression of pDCs, IRF-7mRNA might be involved in the pathogenesis of psoriasis.
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In order to investigate the mRNA expression and function of interleukin-23 (p19/p40) and interleukin-12 (p35/p40) in the psoriatic lesion, no-lesion and normal human skin, reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect the expression of IL-23 (p19/p40) and IL-12 (p35/p40). The results showed that the expression of IL-23p19 mRNA and p40 (IL-12/IL-23) mRNA were higher in psoriatic lesion than those of non-lesional skin and normal skin. The levels of IL-23p19 mRNA and p40 (IL-12/IL-23) mRNA were higher in psoriatic non-lesional skin than normal skin. However, no significant difference was found in the level of IL-12p35 mRNA among the psoriatic lesional skin, non-lesional skin and normal skin. It was suggested that IL-23 might be more important in the pathogenesis of psoriasis than IL-12.
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In order to investigate the correlation between the expression of the apoptotic regulatory proteins (Fas, Bcl-2) in peripheral blood lymphocytes (PBLC) and the level of IFN-gamma and IL-4 in serum of the patients with condyloma acuminata (CA) in the immune pathogenesis of CA, indirect immunofluorescence labeling method of flow cytometer and solid sandwich ELISA method were performed for detecting the expression of Fas, Bcl-2 in PBLC and the level of IFN-gamma and IL-4 in serum of 60 cases of CA. The results showed the expression level of Fas in PBLC of CA was significantly higher than in the normal control group, but the expression level of Bcl-2 was significantly lower (both P<0.01). The level of IFN-gamma in serum of CA was significantly lower than in the normal control group (P<0.01), but IL-4 was significantly lower (both P<0.01). The expression of Fas in PBLC had a negative correlation with the level of IFN-gamma in serum of patients with CA, but had a positive correlation with the level of IL-4; The expression of Bcl-2 had a positive correlation with the level of IFN-gamma, but had a negative correlation with the level of IL-4. All the correlation coefficients had significant difference by t test (P<0.01). It was suggested abnormal apoptosis in PBLC, the suppressed secretion of the TH1-associated cytokines (eg: IFN-gamma) and the increased secretion of the TH2-associated cytokines (eg: IL-4) existed in the patients with CA and might play an important role in the immune pathogenesis of CA.
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Adolescente , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Apoptose , Condiloma Acuminado , Sangue , Citometria de Fluxo , Interferon gama , Sangue , Interleucina-4 , Sangue , Linfócitos , Metabolismo , Proteínas Proto-Oncogênicas c-bcl-2 , Sangue , Receptor fas , SangueRESUMO
In order to investigate the role of murine double minute 2 (MDM2) in cell proliferation and carcinogenesis in condyloma acuminatum (CA), immunohistochemistry and in situ hybridization were used to detect the expression of MDM2 protein and mRNA in normal skin and skin lesions of CA of vulva. PCR was also used to detect HPV types. The results showed that in 32 observed CA specimens, the expression of MDM2 protein and mRNA was detected in 18 (56.25%) and 22 (68.75%) respectively, while the co-expression of MDM2 protein and mRNA was found in 14. PCR results revealed that HPV6/11 and HPV16/18 subtypes were shown in 28(87.5%) and 4 (12.5%) respectively out of 32 CA specimens. Out of the 18 positive specimens expressing MDM2 protein, HPV6/11 subtypes were shown in 15 and HPV16/18 subtypes in 3. In 22 positive specimens expressing MDM2 mRNA, HPV6/11 subtypes were shown in 18 and HPV16/18 subtypes in 4. No expression of MDM2 protein and MDM2 mRNA was observed in normal skin. Our study indicated that the overexpression of MDM2 might be involved in malignant proliferation and carcinogenesis of CA.
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Adolescente , Adulto , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Transformação Celular Neoplásica , Metabolismo , Condiloma Acuminado , Metabolismo , Microbiologia , Patologia , Papillomaviridae , Infecções por Papillomavirus , Metabolismo , Proteínas Proto-Oncogênicas c-mdm2 , Genética , RNA Mensageiro , GenéticaRESUMO
Objective To construct a prokaryotic expression plasmid pET-28a(+) encoding the multiple transferable resistance C (mtrC) gene of N. gonorrhoeae and express it in E.coli DE3, in order to provide a model to study the pathogen's resistance mechanisms to antimicrobial hydrophobic agents. Methods The mtrC gene of N. gonorrhoeae was amplified by polymerase chain reaction from reference strains,cleaved with restriction endonuclease, and then cloned into the prokaryotic expression plasmid pET-28a (+) to construct the recombinant pET-mtrC. This was confirmed by cleavage of restriction endonuclease and DNA sequencing. The recombinant pET-mtrC was transformed into E.coli DE3 to express the protein MtrC with induction by IPTG. Results The mtrC gene in the recombinant pET-mtrC showed 99.5% homology with the reference sequence in GeneBank (U14993). A 48.5 kD fusion protein was identified by SDS-PAGE. Conclusions The successful construction of a prokaryotic plasmid encoding the mtrC gene of N. gonorrhoeae and its expression in E.coli may facilitate the development of a monoclonal antibody to the MtrC protein and help to investigate the mechanism of the mtr efflux system of N. gonorrhoeae.
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The telomerase activity in condyloma acuminatum (CA) tissue with human papillomavirus (HPV) types of 6/11 and 16/18 was detected to investigate the function of telomerase in the occurrence, development and carcinogenesis of genital CA. Forty-two biopsies from patients with genital CA and 30 control tissue samples were tested for telomerase activity, HPV presence and types. The telomerase activity was determined by modified telomerase repeat amplification protocol (TRAP) assay and HPV typing by polymerase chain reaction (PCR) with typing-specific primers. Results showed that HPV-DNA was negative and the expression rate of telomerase was 16.7% in all normal skin samples. All CA samples were positive for HPV (6/11 type was found in 32 cases, 16/18 in 3 and mixed type in 7). Telomerase activity was detectable in all CA patients. The telomerase activity in CA of 16/18 type was apparently higher than in CA of 6/11 type. It was concluded that the hyperplasia in CA might be increased as a result of HPV infection, suggesting that the activation of telomerase by HPV, especially by 16/18 type may play a role in the etiology and carcinogenesis of genital CA.
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Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Condiloma Acuminado , Virologia , Doenças dos Genitais Femininos , Virologia , Doenças dos Genitais Masculinos , Virologia , Papillomaviridae , Classificação , Infecções por Papillomavirus , Virologia , Telomerase , Metabolismo , Infecções Tumorais por Vírus , VirologiaRESUMO
In order to investigate the role of interleukin-4 (IL-4) in experimental murine systemic Candidiasis, we created the intact and dexamethasone-induced immunosuppressed murine systemic Candidiasis models. In these models, two-site ELISA and RT-PCR were applied to determine the level of IL-4 protein and mRNA expression in spleens respectively, clone forming units (CFUs) of infected kidneys were determined with the plating dilution method, and mean survival time (MST) of the mice was recorded. The results showed that, when compared with the controls, protein level of IL-4 increased in both intact mice infected with lethal doses of yeast (day 3, P<0.05; day 7, P<0.001) and immunosuppressed mice infected with sublethal doses of yeast (day 3, P>0.05; day 7, P<0.05). Furthermore, the level of IL-4 was higher on day 7 than on day 3 after infection (P<0.001 and P<0.05 respectively in two groups). The tendency of IL-4mRNA expression was similar with that of IL-4 protein. As for fungal loads in kidneys, CFUs were significantly higher on day 7 than on day 3 after infection (P<0.001 in both groups). Mice in both groups succumbed to infection within several days. It was suggested that IL-4 might play a promoting role in the development of murine systemic Candidiasis.