| Objective1.To investigate the relationship between angiogenic mimicry(VM)and strong penetration long retention(EPR)effects in renal clear cell carcinoma.2.To investigate the relationship between angiogenic mimicry and WHO/ISUP grading in renal clear cell carcinoma.Methods1.Choose 45 patients which been treated between January 2017 to December 2018 in our hospital renal tumor resection and postoperative kidney specimens in vitro in polymer nano micelle contrast agent perfusion experiment,sorts through related data and contains general information such as gender,age,postoperative pathological diagnosis,and polymer nano micelle contrast agent perfusion data measured in the experiment;The peak CT value in the tumor region measured in the perfusion experiment was used to represent the intensity of EPR effect in the tumor region,and the time of reaching the peak of CT value in the tumor region was used to describe the dynamic process of EPR effect in the region.2.Pathological specimens of the above patients were obtained,and then re-sected for HE staining,rabbit monoclonal antibody CD34 combined with Schiff periodate reagent(PAS)staining.According to the results of immunohistochemical staining,the staining results were divided into the experimental group and the control group.The experimental group corresponded to the sections with VM visible under the microscope,while the control group corresponded to the sections without VM.Mimicry vessels were detected and determined,and the positive rate and density of the simulated vessels in all sections were counted and the density was classified.3.Combined with the experimental data and the previous data for statistical and analysis.Results1.In this study,data of 45 subjects were collected and collated completely.Among them,45 cases were pathologically diagnosed as renal clear cell carcinoma,and all underwent in vitro renal polymeric nano-micelle perfusion and imaging measurement after surgery.There were 27 males and 18 females,ranging from 42 years old to 67 years old,with an average of(55.65±7.35)years old.22 tumors were located in the left kidney and 23 in the right kidney.Postoperative pathology confirmed tumor infiltration and fibrous membrane in 37 cases,and no fibrous membrane invasion in 8 cases.The maximum tumor diameter ranged from 2.5cm to 10.5cm,with an average of(6.23±2.10)cm.WHO/ISUP classification included 12 cases of grade 1,8 cases of grade 2,19 cases of grade 2-3,5 cases of grade 3,and 1 case of grade 4.The minimum CT peak value of contrast medium was 2.28×100ug/cm 3,the maximum was9.36×100ug/cm 3,and the average value was(5.58±1.84)×100ug/cm 3.The minimum peak time of contrast medium CT value was 1h,the maximum peak time was 6.5h,and the average peak time was(3.88±1.08)h.2.After tumor specimen sections were stained with HE and CD34 combined with Schiff periodic acid reagent,the staining effect of the sections was ideal under microscope.The sections treated with HE staining showed lumen-like structures surrounded by cancer cells and containing red blood cells.Immunohistochemical staining showed that the lumen structure formed by tumor cells was negative for CD34 and positive for PAS,which was consistent with the criteria of blood VM.The tumor vessels composed of normal vascular endothelium showed positive CD34 and PAS.35 of the 45 subjects were found to be VM positive,corresponding to the experimental group;10 cases were negative for VM,corresponding to the control group.The positive rate of VM in all sections was 77.78%.Mimicry vascular density of 45 sections was counted and scored,including 11 cases with 0score,6 cases with 1 score,8 cases with 2 score,9 cases with 3 score and 11 cases with 4score.3.Chi-square test was used to analyze the correlation between the expression status of VM and general data.When 0.05,the difference was considered statistically significant.χ2=0.536,P=0.464 > between immunohistochemical results and sex;0.05;χ2=0.173,P=0.677 >between VM expression and age;0.05;The χ2 between VM expression and tumor site was0.407,P=0.524 >;0.05;χ 2 =0.043,P=0.835 > between experimental results and tumor envelope invasion;0.05.There was no significant correlation between VM and gender,age,tumor site and capsule invasion.The maximum tumor diameters of the experimental group and the control group were statistically analyzed.The mean maximum tumor diameters of the experimental group and the control group were(6.19 ± 2.20)cm and(6.39 ± 1.83)cm respectively.The t-test analysis showed that there was no significant difference(t=-1.976,P=0.793 >;0.05).The WHO/ISUP grades of the experimental group and the control group were listed and analyzed by rank sum test.The results showed that there was significant statistical difference between the data of the two groups,and the WHO/ISUP grade of the experimental group was higher than that of the control group(U=56.500,Z=-3.412,P < 0.05).4.Combined with the results of immunohistochemical staining and nano-micelle perfusion assay,the average peak CT value of the experimental group was(6.19±1.61)×100ug/cm 3,and the average peak CT value of the control group was(3.44 ± 0.46)×100ug/cm 3.0.01);The average CT peak time of the experimental group was(3.30±1.42)h,and the average CT peak time of the control group was(4.04±0.93)h.The t test was also used to analyze the data of the two groups,and there was no significant difference in the peak time between the two groups(t=-1.976,P=0.055 > 0.05)5.Pearson correlation analysis was performed on the VM score of all slices and the CT peak value obtained from the nano-micelle perfusion experiment,and the correlation coefficient was 0.874,P < 0.05,indicating a strong positive correlation between the two.Pearson correlation analysis was performed on the VM score of sections and the peak time of corresponding CT value,and the correlation coefficient was 0.128,P= 0.401> 0.05.No significant correlation was found between the two.The VM score of sections and the corresponding WHO/ISUP classification were analyzed by Pearson correlation,and the correlation coefficient was 0.685,P < 0.05,indicating a strong positive correlation between the two of them.The peak time of iodine enrichment in VM(+)group was(3.30±1.42)h,and that in VM(-)group was(4.04±0.93)h.There was no difference between the two groups(t=-1.976,P>0.05)Conclusion1.CD34 combined with PAS double staining can effectively distinguish the vascular endothelial vessels and the mimetic vessels formed by tumors in renal clear cell carcinoma tissues,and is suitable for the study of renal clear cell carcinoma tumor microenvironment.2.The expression of VM in renal clear cell carcinoma was positively correlated with the intensity of EPR effect,but not with the peak time of EPR effect.3.The expression of VM in renal clear cell carcinoma was not correlated with age,sex,tumor site,tumor maximum diameter,and capsule infiltration,but positively correlated with WHO/ISUP grade of renal clear cell carcinoma. |
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