Effect of a traditional herbal medicine (shakuyaku-kanzo-to) on testosterone secretion in patients with polycystic ovary syndrome detected by ultrasound
For the quantification of sulfur metabolites in the tissue samples, the tissue metabolites were extracted and derivatized with NEM in ice-cold extraction solvent (80% MeOH: 20% H 2 O containing 25 mM NEM and 10 mM ammonium formate, pH=7.0) which includes stable isotope labeled internal standards (20 M [ 13 C 3 , 15 N]-cysteine-NEM, 36.4 M [ 13 C 2 , 15 N]-GSH-NEM, 10 M [D 4 ]-Cystine, 0.92 M [ 13 C 5 , 15 N 2 ]-GSSG, 20 M [D 4 ]-Hypotaurine, and 20 M of [ 13 C 2 ]-Taurine at a final concentration of 50 mg/mL followed by incubation on 4 C for 24 hr
For NFS1 knock down, the following shRNAs were used: shNFS1_1, TRCN0000229753, B4: shNFS1_2, TRCN0000229755, B6 (Addgene)
instead, they favor moderate doses spread out over time
The remaining 3 studies, scored eight and nine, indicating high-quality studies with low risk of bias (Table 5)
doi: 10.1089/ars.2012.5052 29 HernndezJ