Effects Of ALA-PDT On Expression Of VEGF And CD34 In Condyloma Acuminatum Tissue

Background and Objecktive Condyloma acuminatum(CA)is caused by human papilloma virus(HPV)infection caused by the skin and mucous membrane of the benign vegetation,one of our common sexually transmitted diseases,the pathological manifestations of epidermal keratin The formation of cell hyperplasia,dermal lymphocyte infiltration,capillary hyperplasia and vasodilatation,concave cells is its characteristic performance,distributed in the granular layer and spine layer.Traditional treatment methods are mainly frozen,laser,surgery or topical drugs,effective for warts,but its traumatic and sub-clinical infection is basically ineffective,easy to relapse after treatment.ALA-PDT(5-aminolevulinic acid photodynamic therapy)is widely used in the treatment of genital warts and tumors in recent years,treatment of CA with side effects,low recurrence rate advantage.The basic principle of the photosensitizer specifically in the metabolism of strong tissue accumulation,in a specific wavelength of light,produce singlet oxygen and oxide,which can produce cytotoxic effects and kill the lesion,but no damage to the surrounding normal tissue.However,the mechanism of ALA-PDT to kill diseased tissue is not clear,it may be related to the promotion of apoptosis,inhibition of proliferation,injury of blood vessels,regulation of local immunity and other related,there is still a lack of ALA-PDT on CA angiogenesis.In this study,we investigated the effect of ALA-PDT on the angiogenesis of CA in patients with CA before and after treatment with ALA-PDT,and provided the theoretical basis for the biological mechanism of ALA-PDT to kill CA tissue.Materials and Methods 1.Materials 54 cases of CA patients were from the First Affiliated Hospital of Zhengzhou University dermatology clinic,of which 23 females,31 males,aged 18 to 60 years,mean age 34.15 years;sick course of 19 days-165 days,the average 52.36 days.All patients were eligible for condyloma acuminata laboratory,clinical and pathological diagnostic criteria,the clinical manifestations of patients with typical,pathological diagnosis of clear.All patients were initially diagnosed and did not receive any treatment before the visit,and were excluded from patients with other systemic diseases.2.Methods For the 54 patients who met the experimental criteria,ALA-PDT was treated in all patients before treatment,and the patients were treated with ALA-PDT at 1 week after treatment.The tissue was treated as the experimental group.The positive expression of VEGF protein and CD34 in the CA tissue of the experimental group and the control group were detected by immunohistochemical SP method.Results Criteria: Criteria for VEGF: Positive staining of VEGF protein was light yellow to brown granules,mainly cytoplasmic staining.In the light microscope,the positive cell rate is calculated by randomly extracting each slice of the five different high power field,and then take the average,requiring 400 times the mirror,attached to count 200 cells.Positive cells accounted for the percentage of total cells: 0 points is 0 ~ 10%,1 point is 11% ~ 25%,26% ~ 50% corresponds to 2 points,3 points is 51% ~ 75%,4 points corresponds to 76 ~ 100% The Coloring intensity: 0 points is not coloring,1 minute is light yellow,2 is brown,3 points is brown;the two scores multiplied,the total score.The scores were 0,1 to 4,6 to 8,and 9 to 12,respectively,corresponding to negative(-),weak positive(+),positive(++)and strong positive(+++).Microvessel density(MVD): CD34-positive cells are nuclei or cytoplasm of vascular endothelial cells stained to light brown or yellow particles.First,the high vascular density area was found at(40x)low magnification,and then(200x)high magnification,to find the color of small blood vessels and capillaries and count.Count the number of blood vessels with five high power fields,and the final result is expressed by its average.3.Statistical processing SPSS19.0 statistical software was used to analyze the data.The positive expression rate of VEGF was measured before and after treatment with ALA-PDT by χ2 test.The difference of positive expression intensity before and after treatment was analyzed by rank test.The t test was used to measure ALT-PDT before and after treatment between the two groups MVD differences.Test level using P <0.05 that the difference was statistically significant.Results 1.The expression of VEGF in CA tissue before and after treatment with ALT-PDT The positive staining of VEGF protein was light yellow to brown granules,and cytoplasm was predominant.The positive expression rate of VEGF in 53 cases of CA patients with ALA-PDT was 83.33%(45/54),24 cases were positive after ALA-PDT treatment,and the positive expression rate 44.44%(24/54),the difference was statistically significant(χ2 = 17.69,P <0.001).The expression intensity of VEGF in the warts of ALA-PDT was more than that of ++ ~ +++,and after treatment,the intensity of expression in the ~ ~ ++,the difference was statistically significant(Z =-5.25,P <0.001).2.The changes of MVD in the wart tissue of ALT-PDT before and after treatment In the light microscope to find intravascular blood vessel nucleus or cytoplasm coloring to light to brown or yellow particles that CD34 positive cells.The expression intensity of CD34 in vascular endothelial cells before ALA-PDT was more than that of ++ ~ +++ and MVD is(36.20 ± 8.80)%,The expression of CD34 in vascular endothelial cells after ALA-PDT treatment was more than that of + ~ ++ and MVD(11.83 ± 1.71)%.the difference was statistically significant(Z =-5.25,P <0.001).Conclusion 1.CA expression in ALA-PDT after treatment,the VEGF expression intensity,positive expression rate decreased,indicating that ALA-PDT can inhibit the expression of vascular endothelial growth factor in tissue,and thus inhibit the proliferation of warts,to promote the wart subsided 2.The CA tissue was treated with ALA-PDT with a MVD value measured with a vessel-specific marker CD34 down,indicating that ALA-PDT by reducing the microvessel density in the organization,thereby inhibiting the wart proliferation,promote wart dissipation.3.One of ALA-PDT treatment of CA mechanism is to inhibit the proliferation of vascular endothelial cells and damaged tissue in the new blood vessels.