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Background & objectives: Notably, 17-hydroxy progesterone (17-OHP) (17-OHP) is a precursor for testosterone (T) synthesis, and intratesticular testosterone (ITT) is essential for spermatogenesis. Varicocele (Vx) has an estimated prevalence of 15% in the general population and 35% in those with primary infertility. We aimed to evaluate the correlation between changes of serum 17-OHP after sub-inguinal micro-varicocelectomy and improvement of semen parameters. Patients and methods: The current prospective study included 45 infertile men attending the andrology clinic form February 2021 to August 2021. Two semen analyses and hormonal profile were evaluated. Colored duplex ultasonography (CDUS) was done in standing and supine position for accurate measurements of testicular volumes and confirmation of Vx. Patients underwent sub-inguinal micro-varicocelectomy using a surgical microscope HB surgitech. We followed them prospectively up for three months following micro-varicocelectomy with serum TT and 17-OHP. Results: Sperm concentration improved significantly from 8.36 ± 5.04 million/ml to 12.52 ± 8.42 million/ml after 3 months following sub-inguinal micro-varicocelectomy (p= 0.001), with normalization of concentration in 15/45 (33%) patients. Total motility did not improve significantly but progressive motility improved significantly from 8.62 ± 8.74% to 16.24 ± 14.45% (p=0.001). Abnormal forms significantly declined from 96.67 ± 2.03% to 95.75 ± 2.47% (p=0.009). Serum 17 OHP and 17 OHP/total testosterone (TT) improved significantly from 1.21 ± 0.45 ng/ml and 0.26 ± 0.09 to 1.42 ± 0.76 ng/ml and 0.3 ± 0.16 (p= 0.013, p= 0.004), respectively, while serum TT did not improve significantly. A significant correlation was found between improvement in sperm concentration and both serum 17 OHP and 17 OHP/TT ratio (p=0.001, p=004). Furthermore, change in abnormal sperm forms showed significant correlations with changes in both 17-OHP and 17-OHP/TT. Conclusion: 17 OHP and 17OHP/ TT ratio can be used as biomarkers to detect improvement in semen parameters following sub-inguinal micro-varicocelectomy. [ABSTRACT FROM AUTHOR]