Intracellular Cytokine Expression Profile in Patients with Condyloma Acuminata

Objective: To study the immunological mechanisms ofCondyloma acuminata(CA) through investigating Tlymphocyte subset levels and cytokine profile in theperipheral blood of patients with CondylomaAcuminata. Methods: Tricolor and bicolor immunofluorescentstaining antibody of cell surface antigen and intracel-lular IL-2, IL-4, IL-12, IFN-Y in CD4^+ and CD8^+ T-lymphocytes from 20 patients with CA were performedand followed by flow cytometry. Results: The number of CD3^+T, CD4^+T-lymphocytescells and CD4^+/CD8^+T cells ratio were significantlydecreased(P<0.01) in patients with CA Compared tocontrols, and IL-2, IL-12, IFN-γ production in CD4^+Tcells was decreased(P<0.01), IL-4 and IFN-γ produc-tion in CD4^+T cells was not significantly different (P>0.05), while IL-2 and IL-12 production in CD8^+Tc cellswas decreased (P<0.01), whereas IFN-γ and IL-4 pro-ducing in CD4^+T cells were of no significantlydifference(P>0.05). Conclusions: There was an imbalance of T lympho-cyte subsets, Th1/Th2 cytokines and Tc1/Tc2 in theperipheral blood of CA patients, which may play animportant role in the pathogenesis and progression ofCA.

Correlation between the Expression of Fas, Bcl-2 in Peripheral Blood Lymphocytes and the LeveL of IFN-γ,IL-4 in Serum of Patients with Condyloma Acuminata

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 γ 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 γ 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 γ 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 γ 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 γ, but had a negative correlation with the level of IL 4. All the correlation coefficients had significant differerce by t test ( P< 0.01). It was suggested abnormal apoptosis in PBLC, the suppressed secretion of the THl associated cytokines (eg: IFN γ) and the increased secretion of the TH2 asssociated cytokines (eg: IL 4) existed in the patients with CA and might play an important role in the immune pathogenesis of CA.

DIFFERENTIATION OF PSEUDOCONDYLOMA OF VULVA AND CONDYLOMA ACUMINATA BY DOT BLOT HYBRIDIZATION AND POLYMERASE CHAIN REACTION

This study differentiated pseudocondyloma of vulva from condyloma acuminata using dot blot hybridization and polymerase chain reaction (PCR). A total of 27 cases of pseudocondyloma of vulva and 65 cases of condyloma acuminata were selected for the study. The genital lesions were examined clinically and were biopsied. Each biopsy was subjected to histological examination and HPV DNA analysis by dot blot hybridization and PCR. Dot blot analysis detected HPV DNA in 19(82. 6%) out of 23 cases of condyloma acuminata and 2(25% ) out of 8 cases pseudocondyloma of vulvae (P<0. 05). PCR detected HPV DNA in 51 (92. 7%) out of 55 cases of condyloma acuminata , compared with none in 23 cases of pseudocondyloma (P<0.001 ). HPV DNA was present in the majority of condyloma acuminata specimens. HPV 6 and 11 were the predominant types. Peudocondyloma is probably not associated with HPV. PCR was the most sensitive and useful technique for HPV DNA detection.

COMPARISON BETWEEN IMMUNOFLUORESCENCE AND PCR IN DETECTING HUMAN PAPILLOMA VIRUS IN CONDYLOMA ACUMINATA

Objective To compare the effectiveness of immunofluorescence and polymerase chain reaction （PCR） in detecring human papilloma virus （HPV） in condyloma acuminata （CA）. Methods HPVs in CA tissues from 60 patients were detected by immunofluorescence and PCR, respectively. Different subtypes of HPVs were also identified with restriction fragment length polymorphism （RFLP） . Resuits The positive detective rates of immunofluorescence and PCR were 56. 67 % （34/60） and 96. 67 % （58/ 60）, respectively （ P 〈 0. 01 ）. RFLP results showed HPV6 and HPV11 were the main subtypes in the detected virus, which accounted for 98.28%. Conclusion The sensibility of PCR is superior to that of immunofluorescence.

The Expression of Fas/FasL in Peripheral Blood Lymphocytes and the Level of IL-2 in Serum of Patients with Condyloma Acuminata

In order to investigate the role of the expression of the Fas/FasL in peripheral blood lymphocytes and the level of IL 2 in serum of patients with condyloma acuminata in the immune pathogenesis of CA, flow cytometry, indirect immunofluorescence labeling and ELISA were performed to detect the expression of the apoptotic regulatory proteins Fas/FasL in PBLC and the level of IL 2 in serum of 60 patients with different course of CA. The results showed that the expression of Fas/FasL in PBLC of the group of short course in CA was significantly higher than that of the normal controls ; the expression of Fas, FasL in PBLC of the group of long course CA was significantly higher than that of the group of short course and the normal controls ( P <0.05 and P <0.01, respectively); the level of IL 2 in serum of the group of short and long course CA was significantly lower than that of the group of normal controls ; the negative relation was revealed between the expression of Fas/FasL in PBLC and the level of IL 2 in serum of patients with CA . It was suggested that the abnormal apoptosis in PBLC and decreased level of IL 2 in serum of CA might play an important role in the course of CA.

Etiology and Prevention of Recurrent Condyloma Accuminata

The possible causes of condyloma acuminatarecurrence arc summarized: such as patients with cellularimmune denciencies, physical therapy triggering subclinicalinfective foci, sowing of the virus particles, and insufficienttherapy. Corresponding preventive measures are addressed,including: immunomodulators improving cellular immunity,ensuring the range and depth of physical therapy, and trcatingsexual partners simultaneously.

Evaluation of the Effectiveness of Nd:YAG Laser under Urethroscope in the Treatment of Intraurethral Condyloma Acuminata (CA) in Males

Objective: To evaluate the therapeutic effect of Nd:YAGlaser under urethroscope for the treatment of intraurethralCandyloma Acuminata (CA) in males. Method: Nd:YAG laser under urethroscope was employed toremove the warts. Result: Seventeen patients out of eighteen cases with CAwere cured after the first procedure. The cure rate was 94.4%.For second treatment, the cure rate was 100% and no relapseoccurred after observation for 12-24 months. Conclusion: Nd:YAG laser by urethroscope is a very goodmethod for treatment of intraurethral CA.

TLMA Expression in Condyloma Acuminata from Patients with High and Low Risk HPV

Objective: To detect the activated expression oftelomerase in condyloma acuminata lesions in low-risk (6/11)and high-risk (16/18) human papilloma virus (HPV)infection and examine the role played by telomerase in theoccurrence, development and carcinomatous change ofcondyloma acuminata. Methods: Assaying the expression of telomerase and thetype of HPV in damaged skin of 42 CA patients and normalskin of 30 healthy control individuals through telomeraserepeat amplification protocol (TRAP) and polymerase chainreaction (PCR). Results: In all the normal skin controls, PCR for HPV wasnegative and only 16.7% of samples were positive for TLMAexpression; in CA lesions, HPV testing was positive in all (32cases were low-risk, 3 were high-risk, and 7 were of mixedtype) and all were positive for TLMA expression. Conclusion: TLMA may be activated by HPV infection,and in turn cause the hyperplasia of epidermal cells. It wasalso indicated that HPV, especially high risk types, canactivate TLMA. The activation of TLMA may play animportant role in abnormal hyperplasia and carcinomatouschanges in CA lesions.

EXPRESSION OF MIDKINE IN BENIGN, PREMALIGNANT AND MALIGNANT VULVAR TUMORS

OBJECTIVE: To clarify the role of midkine (MK) in vulvar carcinogenesis though examination of its expression in vulvar lesions including vulvar condyloma acuminata (VCA), vulvar intraepithelial neoplasia (VIN) and vulvar squamous cell carcinomas (VSCC), and to analyze the relationship between MK expression and human papilloma virus (HPV) infection. METHODS: Thirty VSCC, 15 VIN and 10 VCA patients were studied by streptavidin-biotin-immunoperoxidase method. MK expression was compared with clinicopathologic features of vulvar tumors. RESULTS: MK was expressed in 26 of 30 VSCC (87%), 3 of 5 VIN III and all VCA samples, whereas no MK expression was detected in the VIN I-II samples or in normal epithelium. The difference of MK expression between VIN III and VSCC was statistically significant (P

CONSTRUCTION AND IMMUNOGENICITY OF HUMAN PAPILLOMAVIRUS TYPE 6B L1 RECOMBINANT PLASMID

To construct a DNA vaccine as a prophylactic model to prevent condyloma acuminatum and detect its immu-nogenicity in mice. Methods The major capsid protein (L1) gene of human papillomavirus (HPV) 6b was inserted into an eukaryotic ex-pression plasmid (pcDNA3.1). The recombinant plasmid was transfected into COS-7 cells. Western blot were performed to detect whether L1 protein can be expressed in eukaryotic cells. Eighteen female BALB/c mice were tested for immunoge-nicity study. Results The recombinant plasmid (pcDNA3.1-HPV6bL1) was verified as HPV6b L1 gene by sequencing. Western blot showed specific strip. Anti-L1 protein antibodies could be detected in the mice’s sera inoculated with pcDNA3.1-HPV6bL1. Similarly, IL-4, IL-2, and IFN-γ were increased in the same mice. Conclusion HPV6b L1 recombinant plasmid was constructed successfully which had immunogenicity for BALB/c mice. It provided experimental evidence for the research of DNA vaccine of condyloma acuminata..-

Development and optimization of nanoemulsion gel for topical delivery of imiquimod

In the present study,we aimed to formulate,optimize and characterize nanoemulsion-based gel of imiquimod for its topical administration and to improve the drug permeation.Nanoemulsions were prepared by the aqueous phase titration method and spontaneously formed by mixing specific fractions of oil phase:Smix:water.The nanoemulsion formulations were optimized by response surface methodology(RSM) using mixture design,Scheffe model.The formulated nanoemulsion was incorporated into 0.5% Carbopol 934(w/v) to enhance convenience in superficial application of the drug.The nanoemulsions were characterized in terms of droplet size,zeta potential,TEM,DSC and in vitro drug permeation.The vesicle size was 113.6 nm with polydispersity index of 0.251.The zeta potential was 34 m V.The spherical droplet shape was confirmed by TEM analysis.The drug permeation from the diffusion membrane was 73.67% in 6 h for the optimized formulation.An optimized nanoemulsion gel formulation of imiquimod was successfully developed with improved permeation using experimental design technique.The developed formulation could be further explored as a potential alternate to currently available topical formulations for the treatment of genital warts.