1.The Applicable Value Of Serum Inhibin B In Distinguishing Obstructive And Non-obstructive Azoospermia 2.The Study Of Y Chromosome Microdeletions In Patients With Non-obstructive Azoospermia

Currently in the world, the hormone distinguishing OA and NO A with very high sensitivity and specificity does not exit in clinical usage. The need for researching serum INHB has been greater. It has been seventy years when INHB was found. Most trials demonstrated that INHB had good effects in predicting spermatogenesis status and was very promising in clinical applications.The study of INHB is to predict the status of spermatogenesis within the testis. Thus provide the basis for clinical diagnosis and treatment. However, the normal reference range for chinese men as well as the cut point in identifying OA and NOA has not been obtained till now. Because of significant racial differences in serum INHB, the normal reference range as well as the cutoff point in identifying OA and NOA with INHB aborad can not be applied internally. Although some references are given by chinese clinical researcheres, they are not widely used because of small subject number or inferior experimental methods.FSH, seminal plasma neutral a-Glu have differences in patients with OA and NOA. Within recent years an increasing body of evidence has shown that INHB is better than FSH and seminal plasma neutral a-Glu in predicting spermatogenesis.A study can clarify the application values of INHB in distinguishing OA and NOA to provide guidance in theory and experimental evidence for INHB to doctors. The study compared the serum INHB and FSH, seminal plasma neutral a-Glu in forecasting OA and NOA.Objective:This research aims to evaluate the applicable values of serum inhibin B(INHB) in distinguishing obstructive and non-obstructive azoospermia and comparing it with classical parameters including follicle-stimulating hormone(FSH) and seminal plasma neutralα-glucosidase. Meanwhile, to predict the spermatogenetic malfunction in testes. Methods:Semen and blood samples were collected from healthy fertile males(n=60), non-obstructive azoospermia(NOA, n=77) and obstructive azoospermia(OA, n=39) that was defined by testicular biopsy as golden standard for diagnosis. Semen parameters were analyzed. Levels of the serum INHB,FSH and seminal plasma neutralα-glucosidase were determined and their area under curve (AUC), cut-off points, sensitivities and specificities were calculated through receiver operating characteristics(ROC) curve.Results:The 95% confidence interval of serum inhibin B in healthy fertile males was between 20.37 pg/ml and 206.21 pg/ml in our laboratory. There were significant differences in levels of serum inhibin B, FSH, seminal plasma neutralα-glucosidase, INHB/FSH ratio and combination of INHB plus FSH between the OA and NOA(P<0.01), among which, serum INHB alone had the largest AUC of ROC with diagnostic value of 0.985 while the sensitivity and specificity were 97.4% and 92.2% respectively.Conclusion:Serum inhibin B has better sensitivity and specificity in distinguishing OA and NOA than FSH, seminal plasma neutralα-glucosidase, INHB/FSH ratio and combination of INHB plus FSH. Y chromosome microdeletions in loci of azoospermia factor (AZF) will lead to non-obstructive azoospermia(NOA). Therefore, patients with NOA are necessary to be tested. Microdeletion loci can be tested through the PCR technique. However, there have been arguments on the methods and loci in testing Y chromosome microdeletions. Although some researches showed two to twenty-two microdeletion loci situations through single or multi-PCR technique in different males groups, the way on how to measure it have become what we debate.Some researchers have found that patients with NOA had incidence of Y chromosome microdeletions from 1% to 55%. The possible causes of different incidences are given as follows:(1) different selection criteria for subjects; (2) different STSs marker density and locations; (3) different detection methods; (4) different genetic background and environment.Objective:This study compared multi-PCR and single PCR detecting Y chromosome microdeletions, evaluated using 6 or 15 microdeletions loci in testing patients with NOA, so that causes of azoospermia can be verified for clinical diagnosis and treatment in order to provide guidance for clinical practices using this technique.Methods:The 15 classical microdeletion loci were divided into six groups and the optimal multi-PCR reaction conditions in each group and single PCR reaction conditions were found. Then each patient was detected with their own DNA. The multi-PCR and single PCR detection rates,15 and 6 loci microdeletion rates were compared using statistical methods.Results:The six groups'optimal multi-PCR reaction conditions were obtained. Each single PCR reaction condition was explored. In the same 73 patients, statistical significances in either microdeletion patient number and microdeletion locus number (x2=5.06, P＜0.05; x2=15.06, P＜0.05) were found with both traditional 6 loci test and classical 15 loci test. However, no statistically significant (χ2= 2.25, P> 0.05) was found in patient number of microdeletion by comparing single PCR and multi-PCR, while the locus number was statistically significant (χ2=22.04, P<0.05). Among the 12 patients, microdeletion loci at frequency of 4,3,2,1 were detected in 1,2,3,6 patients, respectively.Conclusion:The incidence of 15 microdeletion loci detected by multi-PCR in patients with NOA is 16.44%. Testing 15 Y-chromosome microdeletion loci have much more clinical values than testing 6 microdeletion loci only. Multi-PCR reaction sometimes may lead to false negative results and needs to be verified by single PCR method.The research also found that microdeletion of AZFb+c can lead to very serious damage to spermatogenic function.Y chromosome microdeletion has no relationship to testicular volume.Different PCR machines cannot use the same reaction condition.