| Background and objectives:Human papillomavirus (HPV) are double stranded DNA viruses, which are divided into High risk (LR) and Low risk (LR) genotypes according to a spectrum of anogenital region epithelial proliferative diseases ranging from benign wart to intraepithelial neoplasms associating with HPV. It is now recognized that HR-HPV infection is a necessary cause for cervical intraepithelial neoplasia (CIN) and cervical cancer. LR-HPV6,11are most often detected genotypes in condylomata acuminata (CA) lesions. Epidemiology studies demonstrated that a high percentage of CA lesions contain LR and HR types and HPV16is the most frequently detected HR-HPV type in patients with normal immunity. Two large cohort studies of patients with CA revealed that condylomata acuminata are strongly associated with increased risk of cervical cancer in situ. So we speculate that the infection rate of cervical HR-HPV of females with CA is significantly higher than normal females. Cervical persistent infection of HR-HPV not only might result cervical cancer, but increase transmission chances of virus between sexual partners. Although associations between cervical subclinical infection and cervical cancer have been revealed, there is no effective method to clear HPV in cervix. Topical photodynamic therapy using aminolevulinic acid (ALA-PDT) is a technique for superficial CA with higher recovery rates and lower recurrence rates than the conventional treatments. The main mechanism contributing to the efficacy of ALA-PDT is the phototoxicity to the proliferative cells. In addition, ALA-PDT has been shown to have some antiviral properties. However, there is no research about the anti-HPV property of ALA-PDT by far. The topical application of ALA results in a shallow penetration depth into tissue, which prevents ALA-PDT from sufficiently entering into and destroying multiple or bulky warts lesions. The tissue response varies with the intensity of the cryotherapy ranges from inflammatory response to tissue destruction. Theoretically, the loss or the alteration in the integrity of the tissue structure and the damage of cell constituents resulting from cryotherapy might facilitate ALA penetration int the cells of the wart lesion, which could improve the efficacy of ALA-PDT. The objectives of the study include investigate cervical HR-HPV infection situation in female patients with condylomata acuminate, studying the influence of5-aminolaevulinic acid and photodynamic therapy vs. cryotherapy plus5-aminolaevulinic acid and photodynamic therapy to Hela cells and HPV18E6,E7gene expression, studying the clinical efficacy of5-aminolaevulinic acid and photodynamic therapy for cervical HR-HPV infection and studying cryotherapy plus photodynamic therapy vs. cryotherapy in the treatment of multiple condylomata acuminata.Method:1Study of cervical high risk human papillomavirus infection rate in patients with condylomata acuminata.1.1Cervical exfoliated cells of142patients with CA and212healthy females were detected by hybrid capture II.1.2The positive rates and virus loads of HR-HPV of two groups were compared.2The influence of5-aminolaevulinic acid and photodynamic therapy vs. cryotherapy plus5-aminolaevulinic acid and photodynamic therapy to Hela cells and HPV18E6, E7gene expression.2.1Hela cells were divided into treatment group1(20%ALA-PDT), group2(1%ALA-PDT), group3(0.1%ALA-PDT), group4(0.002%ALA-PDT), group5(cryotherapy plus0.002%ALA-PDT), control group (cryotherapy alone) and blank group (no cryotherapy or ALA-PDT)2.2After12hours of treatment, each group was detected by flow cytometry (FCM). HPV18E6, E7were detected by reverse transcription polymerase chain reaction (RT-PCR) and quantity one software.3Study of the clinical efficacy of5-aminolaevulinic acid and photodynamic therapy treatment for cervical HR-HPV infection.3.1Thirty five patients with cervical HR-HPV infection were divided into ALA-PDT group (n=17) and control group (n=18). A20%ALA solution was applied to the cervix area3h before illumination with red light (635nm,100mW/cm2,100J/cm2).3.2After3month of treatment, cervical HR-HPV of patients in ALA-PDT group and control group were detected. 3.3The negative rates and adverse effects in the two groups were analyzed.4A randomized clinical comparative study of cryotherapy plus photodynamic therapy vs. cryotherapy in the treatment of multiple condylomata acuminata.4.1Eighty patients with multiple CA receive cryotherapy plus ALA-PDT (n=40) or cryotherapy plus placebo-PDT (n=40). After cryotherapy, a20%ALA or a placebo solution was applied to the CA area3h before illumination with red light (635nm,100mW/cm2,100J/cm2). The treatment was repeated7days after the first treatment if the lesions were not completely resolved.4.2The complete response rate, recurrence rate and adverse effects in the two groups were analyzed.Results:1. The positive rates of CA group and control group were54.2%and13.2%, respectively, which was significantly different between two groups (P<0.05). The virus loads (RLU/PC) were divided into four grades. Patients with1～<10grade were24and10,10～<100grade were16and10,100～<1000grade were22and7,>1000grade were15and1in CA group and control group, respectively. The number of patients with high grade virus loads (≥1000) in CA group was significantly higher than that in control group (P<0.05).2. the normal rates, apoptosis rates and dead rates of cells of the seven groups were10.84%,0.23%,86.01%in group1;8.59%,14.83%,74.54%in group2;81.17%,2.80%.8.55%in group3;83.98%,1.66%,11.16%in group4;18.06%,10.80%,67.99%in group5;23.05%,9.40%,58.85%in the control group and88.78%,1.00%,7.63%in the blank group, respectively. The expression rates of E6in group1-5and control group compared with the blank group were0,0,34.02%,49.76%,41.67%, and97.67%, respectively. The expression rates of E7in group1-5and control group compared with the blank group were26.81%,41.20%,61.83%,71.75%,70.30%and93.26%, respectively.3. After one treatments, the negative rates of the ALA-PDT and control group were58.82%(10/17),5.6%(1/18)(p<0.05). The adverse effects in the ALA-PDT group included mild to moderate pain, edema and erosion, without any infection, ulcers, scarring or cervical malformations.4. After two treatments, the complete response rates in the combined group and cryotherapy group were32.4%(36/111) and32.6%(43/132) in the anal area (P>0.05),100%(32/32) and54.5%(18/33) in the urethral meatus (P<0.05), and94.2%(129/137) and50.5%(56/111) in the external genitals (P<0.05), respectively. The recurrence rates in the combined group and cryotherapy group were24.3%(27/111) and31.1%(41/132) in the anal area (P>0.05),9.4%(3/32) and39.4%(13/33) in the urethral meatus (P<0.05), and3.6%(5/137) and31.5%(35/111) in the external genitals (P<0.05), respectively. The adverse effects in each group included mild to moderate pain, edema, erosion and hypopigmentation, without any infection, ulcers, scarring or urethral malformations.Conclusions:1. Detection of cervical HR-HPV in patients with CA should be highly valued because HR-HPV infection rate and virus loads of CA patients were significantly higher than those of healthy females.2. ALA-PDT can inhibit the expression of HPV18E6, E7mRNA in Hela cells; cryotherapy plus ALA-PDT is a more effective regimen for killing Hela cells and inhibiting the expression of HPV18E6,E7mRNA compared with ALA-PDT alone.3. ALA-PDT is a safe and effective therapeutic method for cervical HR-HPV infection.4. Cryotherapy plus ALA-PDT is a more effective regimen for the treatment of multiple CA compared with cryotherapy alone. |
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