Innovative biomedical applications have high requirements for biomedical materials. Herein, a series of biocompatible, antibacterial and hemostatic sponges were successfully fabricated for the treatment of acute upper gastrointestinal bleeding (AUGB). Quaternized chitosan (QC) and soy protein isolate (SPI) were chemically cross-linked to obtain porous SPI/QC sponges (named SQS-n, with n = 30, 40, 50 or 60 corresponding to the weight percentage of the QC content). The chemical composition, physical properties and biological activity of SQS-n were investigated. SQS-n could support the adhesion and proliferation of L929 cells while triggering no obvious blood toxicity. Meanwhile, SQS-n exhibited good broad-spectrum antibacterial activity against both gram-positive bacteria (Staphylococcus aureus) and gram-negative bacteria (Escherichia coli). The in vivo hemostatic effect of SQS-n was evaluated using three different bleeding models. The results revealed that SQS-50 performed best in reducing blood loss and hemostatic time. The overall hemostatic effect of SQS-50 was comparable to that of a commercial gelatin sponge. The enhanced antibacterial and hemostatic activities of SQS-n were mainly attributed to the QC component. In conclusion, this work developed a QC-functionalized hemostatic sponge that is highly desirable for innovative biomedical applications, such as AUGB.