| Objective(1)To explore the levels of vitamin D(25(OH)D)in patients with polycystic ovary syndrome and endometriosis through a systematic review,and to provide evidence-based medicine for clinical practice.(2)To study the relationship between vitamin D level and the clinical pregnancy rate of infertile women receiving IVF/ICSI,and to explore the correlation between vitamin D and ART-related factors,and to use vitamin D and ART-related influencing factors as indicators to predict pregnancy on ART Outcomes,establish a predictive model of clinical pregnancy,guide individualized treatment plans,and improve the success rate of ART.(3)Using bioinformatics methods to further explore the relationship between vitamin D and clinical infertility-related genes,to lay a theoretical basis for the mechanism of vitamin D-related genes in female infertility.Methods(1)Systematic review of vitamin D and clinical infertility-related diseases: computer-searched VIP Full-text Database(VIP),Chinese Journals Full-text Database(CNKI),China Biomedical Literature Database(CBM),China Biomedical Literature Database(CBM),Wan Fang Date knowledge service platform(Wan Fang Date),Pubmed,Embase database with "female infertility","polycystic ovary syndrome","PCOS","endometriosis","vitamin D","25-hydroxyvitamin D","25(OH)D","1,25(OH)2D3","vitamin D" are used as keywords or subject words to search.The inclusion and exclusion criteria and data extraction forms were developed.Two researchers independently screened the literature according to the inclusion and exclusion criteria.The Newcastle-Ottawa Scale(NOS)was used to evaluate the quality of the included literature,and then the required literature was extracted.If there is any objection to the data,quality evaluation and data extraction,it shall be negotiated by both parties or decided by a third party.Meta-analysis was performed using Rev Man 5.2 software,Cochrane Q test was used to qualitatively analyze the heterogeneity of research results,I2 test was used to quantitatively analyze the heterogeneity of research results,and funnel plot,egg and begg test were used to analyze whether the included studies had publication bias,Sensitivity analysis was used to judge the stability and reliability of the results.(2)To construct a clinical model of vitamin D and ART-related influencing factors for predicting pregnancy outcomes: 188 infertile patients who received routine IVF and ICSI treatment for the first time in the reproductive center of our hospital from June 2020 to July 2021 were collected.According to the clinical pregnancy results Divided into successful pregnancy group and pregnancy unsuccessful group.Patient data were collected including vitamin D level,age,BMI(body mass index),years of infertility,type of infertility(primary infertility/secondary infertility),AMH(anti-Müllerian hormone),AFC(Antral follicles),basal sex hormones(blood drawn on 2-4 days of menstruation)including: bFSH(basic follicle stimulating hormone),bLH(basic luteinizing hormone),b E2(basic estrogen),P(basic progesterone),PRL(basal prolactin),T(basal testosterone),DHEA(dehydroepiandrosterone),COH regimen(long regimen/antagonist regimen),Gn dose(total),Gn days,number of pregnancies,HCG day endometrial thickness,HCG day E2(estrogen value),fertilization method(IVF/ICSI),number of oocytes retrieved,number of retrieved embryos,and the observed outcome was clinical pregnancy.(3)Bioinformatics analysis of genes related to vitamin D and clinical infertility: The data related to infertility genes and proteins were downloaded from the NCBI database.DAVID was used to analyze the GO function of the screened differential genes,analyze the KEGG signaling pathway of the differential genes,and import the differential genes into the string database to make a protein interaction network diagram.6.1 Software to screen key genes.Mining modules in the network through the MCODE plugin.Results(1)34 articles were finally included.The meta-analysis showed that serum 25(OH)D and 25(OH)D3 were associated with polycystic ovary syndrome,endometriosis and other unexplained infertility-related diseases.There was basically no publication bias in the 34 literatures on the relationship between them.(2)The independent influencing factors of ART pregnancy outcome in multivariate regression analysis were age of infertility,vitamin D level,AMH,AFC,Gn dose,HCG day endometrial thickness,number of oocytes retrieved,and the area under the ROC curve of the all-dependent variable model was: 75.34%,standard error 0.045,P=0.003,95% confidence interval was 0.712-0.965.Multivariate regression analysis showed that independent factors affecting vitamin levels were identified,including number of embryos harvested,endometrial thickness on HCG day,and clinical pregnancy.(3)10 core genes CYP24A1,CYP27B1,POR,GC,DHCR7,NADSYN1,FABP9,C10orf88 and PTH related to VDR were obtained from GEO.Conclusions(1)Serum levels of 25(OH)D or 25(OH)D3 in women with polycystic ovary syndrome,endometriosis and other unexplained infertility-related diseases are negatively correlated,but the results are not significant.Poor stability.(2)The predictive model established by serum 25(OH)D level and ART-related influencing factors has predictive value for pregnancy outcome.(3)The 10 core genes CYP24A1,CYP27B1,POR,GC,DHCR7,NADSYN1,FABP9,C10orf88 and PTH related to VDR were obtained from GEO,which play a crucial role in the long-term effects of female infertility and health effect.Part One: A Systematic Review of Vitamin D and Clinical Infertility related diseasesObjective Infertility has become a hot issue of medical and social concern.Pelvic inflammation is the common cause,followed by ovulation disorder and immune factors;pelvic inflammation or fallopian tube inflammation caused by fallopian tube obstruction is the main cause.Polycystic ovary syndrome(PCOS)is the main cause of ovulation disorders.Endometriosis,which affects the pelvic microenvironment and causes abnormal immune function,has also become the main factor of infertility after fallopian tube obstruction and polycystic ovary syndrome.Some studies have shown that the deficiency of vitamin D level is related to infertility caused by polycystic ovary syndrome and endometriosis,but there is still a lack of evidence-based medical evidence.The purpose of this study was to explore the level of vitamin D(25(OH)d)in patients with polycystic ovary syndrome and endometriosis through systematic evaluation,and to provide evidence-based medical evidence for this clinical problem.Methods Computer search of the Chinese Journal Database(VIP),Chinese Journal Network Full-text Database(CNKI),China Biomedical Literature Database(CBM),Wan Fang Data Knowledge Service Platform(Wan Fang Date),Pubmed,Embase Data search terms used Including("infertility","polycystic ovary syndrome","endometriosis","PCOS","vitamin D","25 hydroxy vitamin D","25(OH)D","1,25(OH)D3","vitamin D"“female infertility”、“polycystic ovary syndrome”、“endometriosis”.All databases were searched until December 2020.According to the established inclusion and exclusion criteria,two researchers independently selected the literature and extracted the data,used the Newcastle-Ottawa Scal(NOS)to evaluate the quality of the included literature.When there is disagreement with data extraction and quality evaluation,it shall be decided by both parties through consultation or by a third party.Using Rev Man 5.2 software to Meta analysis,Heterogeneity of results was qualitative analysis by Cochrane Q inspection and quantitative analysis by I2 inspection.Funnel plot,Begg test and Egger test were used to check whether there was publication deviation in the included literature.Sensitivity analysis is used to judge the stability and reliability of the results.Results 34 literatures were finally included.The meta-analysis results showed that the serum 25(OH)D25(OH)D3,25(OH)D: [MD=-7.79,95% CI-(-11.63.-3.95)] in women with polycystic ovary syndrome,endometriosis and other unexplained infertility related diseases;25(OH)D3: [MD =-10.65,95% CI=(-14.12,-7.18)],indicating that women with polycystic ovary syndrome,endometriosis and other unexplained infertility related diseases have obvious 25(OH)D or 25(OH)D3 deficiency.However,there are obvious heterogeneity among the studies,among which the index units of vitamin D,detection methods and etiological typing may be the source of some heterogeneity;The results of sensitivity analysis showed that the stability of the results was not good;Funnel plot,egger linear regression and Begg correlation were used to test whether there was publication bias in the included literatures.The results showed that there was no publication bias in 34 literatures on the relationship between serum 25(OH)D,25(OH)D3 and polycystic ovary syndrome,endometriosis and other unexplained infertility related diseases.Conclusion the serum levels of 25(OH)D or 25(OH)D3 in women with polycystic ovary syndrome,endometriosis and other unexplained infertility related diseases are negatively correlated,but the stability of the results is poor,suggesting that due to many influencing factors that have not been effectively controlled,vitamin D deficiency has a certain impact on infertility related diseases,but it does not play an important role,so it should be treated with caution.Part Two: Clinical Model of Clinical Indicators Vitamin D and Related Factors of ART in Predicting Pregnancy OutcomeObiective The purpose of this study was to analyze the different risk factors for clinical pregnancy on ART.To study the relationship between vitamin D level and the clinical pregnancy rate of infertile women receiving IVF/ICSI,explore the correlation between vitamin D and AMH,and use vitamin D and AMH as indicators to predict the pregnancy outcome of ART,and establish a clinical pregnancy prediction model,to guide individualized treatment plans and improve the success rate of ART.Methods From June 2020 to July 2021,188 infertile patients who underwent routine IVF and ICSI treatment for the first time in the reproductive center of our hospital were collected and divided into a successful pregnancy group and an unsuccessful pregnancy group according to clinical pregnancy results.Patient data were collected including vitamin D level,age,BMI(body mass index),years of infertility,type of infertility(primary infertility/secondary infertility),AMH(anti-Müllerian hormone),basal Sex hormones(blood drawn on 2-4 days of menstruation)include: bFSH(basal follicle stimulating hormone),bLH(basal luteinizing hormone),E2(basic estrogen),P(basic progesterone),PRL(basic prolactin),T(basal testosterone),DHEA(dehydroepiandrosterone),COH regimen(long regimen/antagonist regimen),Gn dose(total): including(total FSH + total HMG),Gn days,number of pregnancies,Parity,endometrial thickness on HCG day,E2(estrogen value)on HCG day,fertilization method(IVF/ICSI),number of eggs retrieved,number of embryos retrieved,and the observed outcome was clinical pregnancy.Results(1)Differences in these demographic and clinical characteristics were compared between successful and unsuccessful pregnancy groups.The overall clinical pregnancy rate of clinical pregnancy was 37.8%(71/188).Infertility age,infertility years,vitamin D level,AMH,AFC(antral follicle),bFSH,DHEA,COH regimen,total Gn,number of pregnancies,parity,HCG day endometrial thickness,number of oocytes retrieved were significantly different between the two groups Further logistic regression analysis identified 7 independent factors,including vitamin D level,AMH,AFC,DHEA,number of embryos retrieved,HCG day endometrial thickness,and clinical pregnancy.(2)The risk factors of clinical pregnancy in multivariate regression analysis were infertility age,vitamin D level,AMH,AFC,Gn dose,HCG day endometrial thickness,number of oocytes retrieved,the area under the model ROC curve was 75.34%,the standard error is 0.045,P=0.003,the 95% confidence interval is 0.712～0.965。(3)The clinical pregnancy rate in the vitamin D deficiency group was lower than that in the vitamin D non-deficiency group,and the difference was statistically significant(P<0.05).There were statistically significant differences in membrane thickness and clinical pregnancy(P<0.05).Logistic regression analysis showed that independent factors affecting vitamin levels were identified,including number of embryos retrieved,HCG day endometrial thickness,and clinical pregnancy.Conclusions(1)Infertility age,serum 25(OH)D level,AMH,AFC,Gn dose,number of embryos obtained,and HCG-day endometrial thickness are the influencing factors of clinical pregnancy rate in IVF/ICSI patients.(2)The serum 25(OH)D level was correlated with the number of embryos obtained,endometrial thickness on the day of HCG and clinical pregnancy.(3)The prediction model established by serum 25(OH)D level and ART-related influencing factors has reference significance for the prediction of pregnancy outcome.Part three:Bioinformatics Analysis of Vitamin D and Clinical Infertility Related GenesObiective More and more evidence supports that vitamin D may play an important role in female fertility,but RCT has not determined that vitamin D supplementation plays an important role in assisted reproduction.More and more evidence supports that vitamin D may play an important role in female fertility,but RCT has not determined the impact of vitamin D supplementation on the success of assisted reproduction(Art).Therefore,there is no definite evidence of the relationship between vitamin D and female infertility,and vitamin D may be needed to improve women’s fertility.In this study,bioinformatics methods were used to further explore the relationship between vitamin D and clinical infertility related genes,so as to lay a theoretical basis for the mechanism of vitamin D related genes in female infertility.(Art)the impact of success.Therefore,there is no definite evidence of the relationship between vitamin D and female infertility,and vitamin D may be needed to improve women’s fertility.In this study,bioinformatics methods were used to further explore the relationship between vitamin D and clinical infertility related genes,so as to lay a theoretical basis for the mechanism of vitamin D related genes in female infertility.Methods Infertility gene and protein related data were downloaded from NCBI database.First,enter the GEO homepage,enter the search box on the homepage,enter "female infertility,endometriosis,PCOS",and click search.Look for the infertility gene expression chip "GSE120103" for analysis,save the GEO2 R.txt file,open it with EXCEL,and screen log FC>2 or log FC<-2,P<0.05 as differential genes.Use DAVID to perform GO function analysis on the differential gene after screening,analyze the differential gene KEGG signal pathway,import the differential gene into the sting database to make a protein interaction network diagram,use a gray value greater than or equal to 10 as the standard for screening target genes,and use Cytoscape3.6.1 The software screens key genes.Mining modules in the network through the MCODE plug-in.Results A total of 2371 differential genes were screened,of which 1183 were up-regulated and 1188 genes were down-regulated.Download the matrix file and make a heat map.Using DAVID to perform GO functional analysis of the differential genes after screening,the results show that most of the infertility gene proteins are involved in chemical synaptic transmission: regulation of alternative m RNA splicing;respiratory system development;ossification;skeletal system development;release of sequestered calcium ion into cytosol;regulation of sequestering of calcium ion;hindbrain development;gland development;lung alveolus development;calcium ion transport into cytosol;muscle system process;formation of primary germ layer,etc.Analysis of the differential gene KEGG signaling pathway,the results show that it participates in Neuroactive ligand-receptor interaction with the infertility gene protein: Cytokine-cytokine receptor interaction;Bladder cancer;AGE-RAGE signaling pathway in diabetic complications;Neuroactive ligand-receptor interaction;Hepatitis B;Osteoclast differentiation;Th17 cell differentiation;Chagas disease;Calcium signaling pathway;Chronic myeloid leukemia,etc.,through the MCODE plug-in to explore the modules in the network,The module with the highest score,Cluster1,among which the 10 proteins with the highest scores are CYP24A1(score: 0.946),CYP27B1(score: 0.946),POR(score: 0.925),GC(score: 0.867),DHCR7(score: 0.865),NADSYN1(score: 0.839),VDR(score: 0.815),FABP9(score: 0.800),C10orf88(score: 0.669)and PTH(score: 0.668).Conclusion 10 core genes CYP24A1,CYP27B1,POR,GC,DHCR7,NADSYN1,CYP2R1,FABP9,C10orf88 and PTH around VDR related to VD were obtained from GEO.Several genes have been directly identified that play a crucial role in the long-term effects of female infertility and health. |
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