<p><strong>Objective:</strong> discussion on a series of biospecimen related issues are conducted, such as collection and preservation, quality control, as well as management and application, during the construction of human tissue biobank in a general hospital. <strong>Methods:</strong> To develop a set of standardized operational procedures and to collect tissue samples, such as whole blood, serum, plasma, fresh frozen tissues, cerebrospinal fluid, and paraffin-embedded tissues, which were classified and made aliquots according to different requirements, and stored at -80℃ temperature refrigerator or in liquid nitrogen. At the same time, a set of information management software was used to realize management of the biobank. <strong>Results:</strong> currently, there are more than 20,000 specimens of various benign and malignant cases, which cover 380 diseases, being collected in the biological database in our hospital. These specimens include paraffin-embedded tissue, fresh frozen tissue, femoral head, whole blood, plasma, serum and cerebrospinal fluid, etc. A large number of these specimensare beneficial is used in clinical research at present. <strong>Conclusion:</strong> the establishment of biological sample bank can maximize the value of non-reborn human tissue specimens, and provide normal control standards as well as benign and malignant disease standards for clinical diagnosis and treatment, which is of great significance to the research of disease pathogenesis and the development of detection technology.</p>