The invention relates to the field of 
biopharmaceutics, in particular to a 
liposome preparation for 
anticoagulant thrombolytic difunctional 
fusion protein, namely 
anticoagulant thrombolytic difunctional 
fusion protein of 12 peptides of 
hirudin and reteplase (HV12p-rPA) and a preparation method thereof. The 
anticoagulant thrombolytic difunctional 
fusion protein (HV12p-rPA) constructed in the laboratory has the dual effects of anticoagulation and 
thrombolysis by structural identification, expression, 
chromatography renaturation, purification and extracorporal and intracorporal pharmacodynamic experiments. In the preparation method, the 
liposome preparation is prepared from the anticoagulant thrombolytic difunctional fusion 
protein serving as a 
raw material, and a 
liposome is prepared by an optimized membrane dispersion-probe ultrasonic method; and the optimized membrane dispersion-probe ultrasonic method is characterized in that a prescription which is most suitable for improving the envelop rate of the HV12p-rPA liposome is selected by taking a ratio of 
phospholipid to 
cholesterol, the concentration of 
protein medicaments, the volume of 
buffer solution and water-bath ultrasonic time as influence factors, and the grain 
diameter of the HV12p-rPA liposome is reduced and homogenized further by probe ultrasonic, so that the HV12p-rPA liposome of which the grain 
diameter is between 140 and 145 nanometers and which is used for intravenous injection is prepared. By the preparation method, the envelop rate of the prepared HV12p-rPA liposome is over 90 percent, and extracorporal thrombolytic activity and extracorporal 
anticoagulant activity are 23,810 
international unit (IU) .mg<1> and 414 
antithrombin unit (ATU) .mg<1> respectively.