RESUMO
Objective:To establish a new molecular typing method for Treponema pallidum (TP) based on TP0136 protein sequence heterogeneity. Methods:The amino acid sequences of TP0136 open reading frame (ORF) of 9 strains of Treponema pallidum ssp. Pallidum (TPA) , 3 strains of Treponema pallidum ssp. Pertenue (TPE) , 1 unclassified simian strain of Treponema Fribourg-Blanc (FB) and 1 strain of Treponema pallidum ssp. Endemicum (TEN) were searched from Genbank, and multiple sequence comparisons were performed to obtain the molecular typing results of TP0136 protein. The TP0136 protein-based molecular typing method was used to classify 23 TPA clinical isolates, which were collected from Dermatology Hospital of Southern Medical University from January 2015 to December 2018, and the typing results were compared with those by the traditional typing method based on the tp0548/Arp/Tpr genes. Results:TP0136 protein was highly heterogeneous in different TP strains. According to the amino acid sequence of TP0136, TPE, FB and TEN strains were divided into 4 subtypes of Ⅰ- Ⅳ, TPA strains were divided into 6 subtypes of Ⅴ-Ⅹ, and TPA clinical strains were classified into 4 subtypes of Ⅶ, Ⅸ, Ⅹ, Ⅺ. Through the traditional typing method described above, 23 TPA clinical strains could be divided into 5 types (13D/d, 14D/f, 14D/g, 15D/f, 16A/e) . By using the TP0136 protein-based typing method combined with traditional typing method, the above clinical strains could be further subdivided into 10 types, and the 14D/f type could be further divided into 3 subtypes by using the TP0136 protein-based typing method.Conclusion:The TP0136 protein-based molecular typing method can be used to distinguish TP species, which is helpful for further improvement of traditional TPA molecular typing.
RESUMO
Objective To evaluate the effects of extracts of Semen Coicis (ESC) on a BALB/c mouse model of atopic dermatitis (AD),and to explore its potential mechanism.Methods Forty specific pathogen-free (SPF) female BABL/c mice were randomly divided into blank group (8 mice,receiving no treatment) and AD model group (32 mice).The mice in the model group were topically treated with 2,4-dinitrochlorobenzene (DNCB) in acetone/olive oil to establish the mouse model of AD.After modeling,8 mice in the blank group and 8 in the model group were sacrificed immediately.The other 24 mice in the model group were randomly and equally divided into 3 groups:model control group receiving no treatment,ESC group and ESC vehicle group topically treated with ESC and ESC vehicle respectively once every day on the back and aural region of the mice for 28 consecutive days.Changes in skin lesions were observed by naked eyes every day.A thickness tester was used to measure the thickness of skin lesions on the left ear before modeling,at completion of modeling and 12 hours after the final treatment.At 12 hours after the final treatment,the mice in the above 3 groups were sacrificed,and the eyeballs were removed for collecting blood.Then,the sera were isolated,and skin tissue specimens were obtained from the skin lesions on the back.These tissue sections were subjected to hematoxylin and eosin (HE) staining and toluidine blue staining for observing the infiltration of inflammatory cells in skin lesions.An immunohistochemical study was performed to determine the expression of aquaporin 3 (AQP3),Toll-like receptor 2 (TLR2) and TLR4,and enzyme-linked immunosorbent assay (ELISA) to detect the serum levels of IgE,interleukin-4 (IL-4) and interferon-/ (IFN-γ).Results After 28-day treatment,skin lesions were improved in the ESC group.Compared with the model control group,the ESC group showed a significantly lower clinical symptom score (1.50 ± 0.58 vs.2.50 ± 0.58,P ＜ 0.05),decreased lesional thickness on the left ear ([0.31 ± 0.01] mm vs.[0.33 ± 0.01] mm,P ＜ 0.05),and lower number of infiltrating mast cells per high-power field (15.18 ± 1.64 vs.28.94 ± 1.28,P ＜ 0.05).Immunohistochemical findings indicated that the ESC group showed significantly lower expression of AQP3,TLR2 and TLR4 compared with the model control group,and decreased AQP3 expression in the spinous layer.Compared with the model control group,the ESC group showed significantly lower total serum IgE and IL-4 levels,but higher IFN-γ levels (all P ＜ 0.05).Conclusion Topical ESC is effective for the treatment of skin lesions in mouse models of AD,likely by regulating serum levels of IgE,IL-4 and IFN-γ and affecting the expression of AQP3,TLR2 and TLR4.
RESUMO
Objective To observe the effect of glycyrrhizin (CG) on the proliferation and the expressions of interleukin (IL)-2, IL-4 in human keratinocytes cell line HaCaT cells, and to investigate the therapeutic mechanism of CG in the pa?tients with psoriasis. Methods The HaCaT cells were divided into four groups:blank group (without CG), low concentration group (50μL CG/mL cell supernatant), medium concentration group (100μL CG/mL cell supernatant) and high concentra?tion group (200μL CG/mL cell supernatant). Methyl thiazolyl tetrazolium (MTT) assay was performed to evaluate the prolifer?ation of HaCaT cells that were treated with CG at different concentrations respectively for 24 hours. The contents of IL-2 and IL-4 in HaCaT cells were examined by enzyme linked immunosorbent assay (ELISA). Results Compared with the blank group, there were obviously inhibitory effects in the proliferation of HaCaT cells treated by low,medium and high concentra?tions of CG for 24 hours. There was no significant difference in IL-2 level between low concentration CG group and blank group (ng/L:234.51±17.98 vs 225.31±16.23, P>0.05). After 24 hours of intervention with CG of medium and high concen?trations, the expression levels of IL-2 was significantly lower in HaCaT cells than that in blank group and low concentration group. The IL-2 level was significantly lower in high concentration group than that in medium concentration group (ng/L:188.99±19.22 vs 208.49±18.40, P<0.05). At the same condition, the secretion of IL-4 in HaCaT cells was significantly up-regulated in high concentration group than that in other three groups (ng/L:45.67±10.29 vs 37.62±3.90, 39.68±6.08, 43.85± 10.26, P<0.05). Conclusion Glycyrrhizin may suppress the proliferation of human skin keratinocytes by regulating secre?tion of cytokines IL-2 and IL-4.
RESUMO
Objective To assess polymerase chain reaction(PCR)combined with restriction fragment length polymorphism(RFLP)and gene sequencing technologies in the detection and typing of HPV DNA.Methods Tissue specimens were collected from skin diseases and venereal disease in perianal or genitals.PCR was performed with HPV DNA general primers(MY09/11)in tissue samples. Positive fragments of HPV DNA were purified and digested by restriction enzymes.The digested fragments were typed by po]yacrylamide gel electrophoresis(PAGE).The Resultswere verified by direct sequencing.Results In 50 clinical samples there were 35 HPV DNA positive,including 26 from patients with condyloma acuminatum,8 from patients with bowenoid papulosis,and 1 from patients with squamous cell carcinoma.In HPV DNA positive samples,19 were HPV6,3 were HPV11,8 were HPV16,4 were HPV6 and HPV 11,and I was HPV62.Sequencing Resultswere in accordance with the PCR-RFLP Results .Conclusion PCRRFLP method is effective in the detection and typing of HPV DNA.
RESUMO
Objective To investigate the influence of an anti-integrin α6 monoclonal antibody (GoH3) on the in vitro infection of a human keratinocyte cell line HaCaT with HPV6/11 virus particles (VP). Methods HaCaT cells were infected in vitro with 4 different concentrations of HPV6/11 VP alone, HPV6/11 VP of 106 copies/ml after incubation with 6 different dilutions of GoH3, or 8 clinical isolates of HPV6/11 VP of 106 copies/ml before or after the incubation with 1∶ 100 dilution of GoH3. After additional culture, the infected HaCaT cells were collected and fluorescence quantitative (FQ)-PCR was performed to detect the HPV DNA load in cells. The inhibition rate of CoH3 on the infection was calculated. Results The viral load was different among the HaCaT cells infected with different concentrations of HPV6/11 VP (P < 0.01). The inhibition rate on the infection positively correlated with the concentration of CoH3 when the dilution was more than 1∶ 100; however, when the dilution was less than 1∶ 100, the increase in CoH3 concentration had no influence on the inhibition rate. The average viral load in HaCaT cells infected with clinical isolates of HPV6/11 VP was (5.81 ± 2.51) × 104 copies/ml in the absence of GoH3, (3.02 ± 1.21) × 104 copies/ml with the presence of CoH3, and the average inhibition rate of GoH3 was (46.9 ± 4.7)%. Conclusions GoH3 could partially suppress the adhesion of HPV6/11 VP to HaCaT cells, hinting that integrin a6 is an important HPV6/11 VP receptor on host cells.
RESUMO
Objective To investigate the expression of integrin α6 mRNA in skin lesions of patients with condyloma acuminata. Methods Sixty outpatients with condyloma acuminate, including 30 primary cases and 30 recurrent cases were enrolled in the study;30 non-condyloma acuminate patients undergoing circumcision were used as normal controls. The expression of integrin α6 mRNA in condyloma acuminata lesions of primary and recurrent cases, as well as in skin samples of normal controls were determined by RTPCR. ANOVA was used to compare the differences among the groups, and Student-Newman-Keuls test(q test)was used for pairwise comparisons. Results Relative contents of integrin α6 mRNA(integrin α6/GAPDH)in normal controls, primary and recurrent condyloma acuminata lesions were 0.25±0.10, 0.79±0.16 and 1.07±0.29, respectively, and the difference was of statistical significance(F=127.687, P=0.001). Conclusion Integrin α6 may be associated with the pathogenesis and recurrence of condyloma acuminate, which may provide a new target for prevention and treatment of the disease.
RESUMO
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.