| Objective To explore the predictive value of endometrial CD 138 immunohistochemical examination on pregnancy outcome of in vitro fertilization-embryo transfer(IVF-ET),and to provide a reference for threshold setting of antibiotic treatment in chronic endometritis patients before assisted reproduction.Methods A total of 193 eligible patients who underwent IVF-ET and endometrial biopsy in reproductive medicine center of Zhejiang Provincial People’s Hospital from December 2015 to December 2019 were selected as the study subjects.The endometrial tissue wax block was taken out for CD138 immunohistochemical staining.A cell was considered as CD138+if its plasma exhibited basophilia and complete brown staining,with an intact cell membrane and a clear nucleus,and appeared as individual cells or as small clusters of cells,which represent for plasma cell.The CD138+cells counting was assisted by a digital pathology slide scanner with annotation platform,then Image J was used to calculate the tissue area of each slice.In this study,the total number,and the average density of CD 138+cell per slice were used as two quantitative methods for plasma cells counting.The biochemical pregnancy rate,embryo implantation rate,clinical pregnancy rate and live birth rate were regarded as pregnancy outcome indicators.According to whether or not be clinical pregnancy,the patients were divided into two groups,the total number of CD138+cells and the density of CD138+cells between the two groups were compared.According to whether or not presence of CD138+cells,the patients were divided into two groups,pregnancy outcomes between the two groups were compared.Logistic regression analysis was performed on various factors related to clinical pregnancy,including the demographic characteristics of patients,the number and the type of embryos transferred,endometrial thickness and CD 13 8+cell number,to screen the independent significant factors and verify the significant effects of CD 13 8+cell on clinical pregnancy by two different counting approaches.Reference to clinical pregnancy,two receiver operating characteristic curves(ROC)were drawn according to two plasma cell quantitative methods,respectively.Then,divided by the best optimal cut-off points,the patients were grouped and pregnancy outcomes,rate were compared,respectively,to judge the predictive ability of the two quantitative methods for pregnancy outcomes.According to the hierarchical density level of CD138+ cells,the patients were further divided into 0/HPF,(>0-1)/30HPF group,(>13)/30HPF group and>3/30/HPF group,various pregnancy outcomes were compared among these groups.Results 1.The total number and the density of CD138+ cells in the clinical pregnancy group were statistically significantly higher than those of the group without clinical pregnancy(P=0.005,P=0.003).2.The CD138+ group was consisted of 104 patients(53.89%),while the CD138+ group was consisted of 89 patients(46.11%).The biochemical pregnancy rate,embryo implantation rate,clinical pregnancy rate and live birth rate of these two group were 17.31%vs 8.99%,34.54%vs 52.94%,45.19%vs 64.04%,32.69%vs 52.81%,respectively,with statistical significance in the embryo implantation rate,clinical pregnancy rate and live birth rate(P=0.01,P<0.001,P=0.005).3.Logistic regression analysis of clinical related factors showed that both the number and density of CD138+cells were independent factors for clinical pregnancy,and the patients without any CD138+ cell were 2.48 times more likely to have a successful pregnancy than those with CD 138+cells.4.The area under the ROC of the whole slice total number of CD138+ cells were 0.612,and its best optimal cut-off point was 6.50/slice with sensitivity of 88.5%and specificity of 31.5%.The area under the ROC curve of whole slice density of CD138+ cells was 0.618,and its best optimal cut-off point was 1.20/30HPF with sensitivity of 78.8%and specificity of 42.7%.Divided by 6.50/slice,the patients were grouped into group 1A(total number of CD138+cell ≤6.50/slice)with 153 patients(79.27%)and group 1B(total number of CD138+cell>6.50/slice)with 40 patients(20.73%).The biochemical pregnancy rate,embryo implantation rate,clinical pregnancy rate and live birth rate of the two groups were 10.45%vs 25.00%,48.44%vs 22.58%,60.13%vs 30.00%,48.67%vs 20.00%,respectively,all with statistical significancy(P=0.016,P<0.001,P=0.001,P=0.001).Divided by 1.20/30HPF,the patients were grouped into group 2A(density of CD138+ cell ≤ 1.20/30HPF,133(68.91%)patients),and group 2B(density of CD138+ cell>1.20/30HPF,60(31.09%)patients).The biochemical pregnancy rate,embryo implantation rate,clinical pregnancy rate and live birth rate of the two groups were 9.02%vs 23.33%,48.67%vs 30.43%,61.65%vs 36.67%,47.37%vs 31.67%,respectively,all with statistical significancy(P<0.001,P=0.003,P=0.001,P=0.041).5.Patients were divided into four groups according to the average density of CD138+cells:0/HPF(group 3A),(>0-1)/30HPF(group 3B),(>1-3)/30HPF(group 3C)and>3/30HPF(group 3D).The biochemical pregnancy rate of the four groups were 8.99%,10.26%,16.67%and 25.71%,respectively,showing an upward trend(P=0.084).Comparison between groups revealed the difference between group 3 A and 3D had statistical significancy(P=0.015).The embryo implantation rates of the four groups were 52.94%,37.50%,35.42%and 30.19%,respectively,with a significant downward trend(P=0.015),and the difference of group 3 A compared to group 3B,3C and 3D all had statistically significant(P=0.038,P=0.034,P=0.004).The clinical pregnancy rate of the four groups were 64.04%,51.28%,43.33%and 37.14%,respectively,with a significant downward trend(P=0.028),and the difference of group 3A compared to group 3C and 3D had statistically significant(P=0.046,P=0.007).The live birth rate of the four groups were 52.81%,35.90%,33.33%and 28.57%,respectively,with a significant downward trend(P=0.039),the difference between group 3A and 3D had statistical significancy(P=0.015).Conclusion 1.The number of endometrial CD138+cell is associated with outcome of IVF-ET.The higher the total number and density of CD138+ cell,the lower the rate of clinical pregnancy,embryo implantation and live birth,and the higher the rate of biochemical pregnancy.2.The number of endometrial CD138+ cell has good sensitivity but poor specificity for predicting pregnancy outcome.3.A CD138+cell density up to 1/30HPF can negatively affect IVF-ET pregnancy outcome. |
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