| Background and ObjecktiveCondyloma acuminatum(CA)is a benign neoplasms of the skin and mucous membrane caused by human papillomavirus(HPV)infection,which often grows in the parts of the body such as the external genitalia,anal regions,urethra and cervix,very few in the toes,mouth,umbilical fossa and so on.In recent years,its incidence continues to rise,and is one of the very common sexually transmitted disease(STD)of our country,and this disease is mainly transmitted through sexual contact,sexually active young men and wonmen are mainly infected objects.Although CA is a benign proliferative but it has proliferates rapidly,highly contagious and has a high recurrence rate,and some large cases of CA may occur malignant.Patients have a greater psychological stress,even some patients with depressive symptoms,seriously affecting the physical and mental health of the patients.CA has become one of the key detection of STD by the Centers for Disease Control(CDC).CA is a benign and abnormal proliferative disease,however,it,proliferates rapidly,highly contagious and has a high recurrence rate,which is related to the abnormal proliferation and abnormal apoptosis of keratinocytes in CA tissue.But the exact mechanism of abnormal proliferation and abnormal apoptosis of keratinocytes in CA tissues is not very clear,which may be associated with a variety of apoptosis-related genes.The apoptotic protein Bad is a member of the Bcl-2 family of proteins,which have BH3 homology domains and sequences,can promote apoptosis.When the apoptosis stimulus signal is present,the structure of Bad has changed,which translocates immediately to the mitochondrial outer membrane and binds Bcl-xL after its activation,inducing apoptosis.A number of studies have shown that pro-apoptotic protein Bad is of great significance to the formation and metastasis of various tumors.PCNA,as a coenzyme of DNA polymerase,is an acidic nucleoprotein that can participate in the replication of DNA during abnormal proliferation of cell,which is closely related to the regulation of cell cycle,PCNA is a recognized marker of the cell S phase of tumor and often widely used in cell abnormal hyperplasia of many experimental tests.However,there is a lack of research on the expression of pro-apoptotic protein Bad and its association with PCNA in CA tissue.In order to explore the expression of Bad and their relationship with PCNA in CA tissue,this study detect the expression of Bad and PCNA in CA and normal foreskin by the immunohistochemical method to explore the possible pathogenesis of the occurrence and development of CA by Bad and PCNA.Materials and Methods1.MaterialsA total of 53 patients were included in the study.Among these 53 patients,32 were male and 21 were female.Patients were 16~65 years old,with a mean of 32.5+8.7 years old.The course of CA was 14~180 days,and the average range was 50.7+4.3 days.CA patients in the experimental group were treated in the clinic of skin and STD of the first affiliated hospital of Zhengzhou University from September 2015 to September 2016.All patients had typical clinical manifestations of CA,Vinegar white test positive,HPV infection positive,and the histopathology of lesions is consistent with the histological manifestations of CA,which were associated with the CA standard,without other systemic diseases.Patients with skin lesions were not received any treatment,such as immune drugs,freezing,laser and any other related treatment.The experimental specimens of control group is the foreskin tissue of 28 cases of normal adult male skin after resection in our hospital urology,and the foreskin tissue was confirmed by histopathological examination as normal tissue epithelium.2.MethodsThe expression of Bad and PCNA were detected by immunohistochemistry.All experimental steps are carried out in strict accordance with kit instructions.The positive expression of Bad was mainly detected in the cell membrane and cytoplasm,the colour of positive particles was brownish-yellow to brown.PCNA was positively expressed mainly in the cell nucleus,together with brown particles.The scoring criteria was based on the quotationed literature,and according to the tinting strength and percentage of positive cells in the tissue section,and quantitative classification was performed.The tinting strength was scored as follows: no colour,0 points;light yellow,1 point;brown-yellow,2 points;brown,3 points.The percentage of positive cells was scored as follows: <10%,0 points;11%~25%,1 point;26~50%,2 points;51%~75%,3 points;?76%,4 points.Taking the product of the above two point systems as the total number of points,the final scores were interpreted as follows: strongly positive(+++),9~12 points;positive(++),6~8 points;weakly positive(+),1~4 points;negative,0 points.3.Statistical treatmentThe experimental data were analyzed statistically by SPSS17.0 statistical software,The positive expression rate of Bad and PCNA was compared by ?2 test;the positive expression intensity of Bad and PCNA used the rank sum test method;The relationship between Bad and PCNA in CA tissue was analyzed by Spearman method in non-parametric system.The standard of examination was P <0.05,the difference was statistically significant.Results1.The Bad expression in CA and the normal foreskin tissuesAmong the 53 CA patients,the positive expression rate of Bad was 69.81%(37/53),the positive expression intensity was ++~+++,and the positive color was mainly observed in the cell membranes and cytoplasm;In the normal tissue samples(n = 28),the positive expression rate of Bad was 28.57%(8/28),and the positive expression intensity was-~+.The positive expression rate of Bad in these two groups was significantly different(?2 = 18.436,P <0.05),and the expression intensity of Bad was significantly significant between these two groups(H= 23.063,P <0.05).2.The PCNA expression in CA and normal prepuce tissuesAmong the 53 cases in the CA group,PCNA was positively expressed in 46 cases(86.79%),the expression intensity ranged from ++ ~+++,and positive colors are mainly located in the nucleus,most of them are brown granules;In the 28 cases of the control group,the positive expression rate of PCNA was 39.28%,the positive expression intensity was +~++.The positive expression rate of PCNA in the two groups was statistically different(?2 =4.356,P <0.05),and the positive expression intensity of PCNA was statistically significant between these two groups(H = 13.452,P <0.05).3.Correlation analysis of Bad and PCNA expression in the CA tissuesAmong 37 cases with positive Bad expression,31 cases were positive of PCNA and 6 were negative;In 46 cases with positive PCNA protein expression,Bad expression was positive in 31 cases,negative in 15 cases.A Spearman rank correlation analysis indicated that Bad expression was positively correlated with PCNA expression(r = 0.379,P < 0.05).Conclusions1.The positive expression rate and positive expression intensity of Bad in CA tissues is higher than those in the control group,indicating that Bad was excessive overexpression in CA tissues,which may promote the apoptosis of keratinocytes in CA tissue,to a certain extent,and inhibit the hyperplasia of CA tissue.2.The positive expression rate and positive expression intensity of PCNA in CA tissues were higher than the control group,showing there is a high expression of PCNA in CA tissue,suggesting that CA tissue had abnormal proliferation of keratinocytes.3.There was a positive correlation between Bad and PCNA expression in CA tissues,suggesting that Bad and PCNA may be involved in the germination and development of CA,and showing CA is regulated by a variety of apoptosis-related genes,but its exact mechanism still needs further study. |
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