An experiment was conducted to study the effects of yeast selenium (SeYst), chromium picolinate (CrPic) and their interaction on performance, lipid metabolism and egg quality of laying hens. Three hundred and twenty 45-week-old H-Sex hens were randomly assigned to four treatments with four replicates each treatment and twenty birds in each replicate. Four dietary treatments were as followings: 1) basal diet with no chromium supplementation (group Ⅰ); 2) basal diet+0.4 mg/kg CrPic (group Ⅱ); 3) basal diet+0.5 mg/kg SeYst (group Ⅲ); 4) basal diet+04 mg/kg CrPic+05 mg/kg SeYst (group Ⅳ), respectively. The trial lasted for 35 days. The results showed that: 1) laying rate in groups Ⅲ and Ⅳ was significantly increased and ratio of feed to egg was reduced (P<0.01); 2) serum cholesterol contents in groups Ⅱ and Ⅳ were significantly reduced (P<0.01), and serum HDL-C contents in groups Ⅲ and Ⅳ were significantly increased (P<0.01); and yolk cholesterol contents in groups Ⅱ, Ⅲ and Ⅳ were significantly reduced compared with those in group Ⅰ (P<0.01); 3) egg selenium contents in groups Ⅲ and Ⅳ were significantly increased compared with those in groups Ⅰ and Ⅱ (P<0.01); 4) egg selenium contents in groups Ⅲ and Ⅳ were increased compared with those in group Ⅰ from starting to 2nd week, and those in groups Ⅰ-Ⅳ reached a relatively steady value from 2rd to 5th week of the trial. These results indicated that supplementation of CrPic reduced cholesterol contents in serum and yolk, and supplementation of SeYst reduced ratio of feed to egg and yolk cholesterol content and increased laying rate and egg selenium content. Combination of SeYst and CrPic increased laying rate, and reduced ratio of feed to egg and cholesterol contents in serum and yolk, and increasd serum HDL-C content and egg selenium content.