The Focus Ultrasound On Persisitent Infec Tion Of HR-HPV: The Rapeutic Mtchanism And Clinical Preliminary Study

Uterine cervix cancer is the most common tumbor in the female. Persistent infection of high risk human papiloman virus (HR-HPV) is the nitiating agent for cervical intra-epithelial neoplasia (CIN) and cervical cancer,Treating the persistent infection of HR-HPV has became a very important means to prevent and cute the cervical cancer. However there was no effective treatment for it. As a new technique for cervical disease, whether can focus ultrasound interdict the persistent infection of HR-HPV. In this study, through to research the effection of Focus ultrasound on expression of HPV E6 and E7 gene of the Caski cells, we can see the therapeutic mechanisms to interdict the persistent infection of HR-HPV, and we adopt a double-blind comparison research to study the clinical efficacy of focus ultrasound for HR-HPV infection.Part1 The effection of diffrrent dose of focus ultrasound for the morphologic changes and the survival rate of Caski cellsAfter irradiated with different time including 10,20,30,40,50S by a fixed exposure intensity of focus ultrasound(The focal length of 4-6 mm, operating at a frequency of 10.2 MHz and a acoustic power of 3.5W).The morphologic changes of Caski cells were examined with microscop.The cell survival rate was measured by Methyl Thazolyl Tetrazolium (MTT) assay.Results⑴Along with prolongation of irradiate time,caski cells losed the nomal shape and became accrescent and round. At the same time,The cellular nucleus of caski cells was accrescent,with their nuclear material became greater and more obvious.⑵The result of MTT assay showed: After 24 hours by irradiated with focus ultrasound for 30S and 50S,the suviver rate of Caski cell was 55.7% and 26.1% respectivly.Along with the lengthen of irradiated time, the surviver rate of caski cells had became gradually reduced,and the difference has statistical significance(P<0.05).Conclution The Fucos ultrasound can impair the Caski cells visibly,the longer of irradiated time,the more significant of cells anormal forms and the lower of survive rate of cells.Part 2 The effection of focus ultrasound for HPV E6 E7gene in Caski cellsAfter irradiated for 30s and 50s respectively, the cells was incubationed for 24 hours. We extracted the con-RNA of Caski cells using Trizol kit. The expressions of HPV16 E6,E7 genes were simultaneously detected by reverse transcription - polymerase chain reaction (RT-PCR) . Then take an electrophoresis in sepharose whose saturation was 1%. Observe the results were analysised by image analysis systerm. Their optical density (OD) was examined wih ultraviolet spectrophotometer.Results The mRNA expressions of HPV E6 E7 were decreased after irradiated by Focus ultrasound for 30S, And it was the lowest after irradiated for 50S. The ration of detected strap toβ-actin strap was caculated.We compare their optical density and the difference has statistical significance(P<0.01)Conclution Focus ultrasound can inhibit the expression of HPV16 E6E7 genes effectively.Part 3 The clinical preliminary study of focus ultrasound for cervical HPV infectionA total of 90 women with cervical high-risk HPV infection were divided randomly into ultrasound therapy group(30cases),interferon therapy group (30cases) and control group (30cases) . In ultrasound therapy group, the patient were treated by the ultrasound therapeutic device, which was developed by the Chongqing Haifu(HIFU) Technology,Co.Ltd,China. In interferon therapy group,the patients were injected theα-2b interferon 3 million unit on the cervical 12,3,6,9 point,every other day,the frequency total 10. After 3 months and 6 months'follow up, the tite of HPVDNA were detected by Hybrid Capture 2, and the negative rate of HPV infection compared in three groups,. Results In the third month,the negative rate of ultrasound group were 53.3%;And it was 48.3% for interferon treatment;To control group,it was 31.0%. In the sixth month,the negative,remission,and ineffective rate of ultrasound group changed into 82.8%,And it was 79.3% for interferon treatment;To control group,It was 53.6%. There was highly significant statistical difference in the three groups (P< 0.05),the negative rate of two therapy groups were obviously higher than the control group, however it was insignificant difference between the ultrasound group and interferon group (P>0.05).Conclusion The ultrasound treatment can control the infection of cervical HR-HPV effectively.