Objective: To investigate the significance of using negative pressure biopsy in the diagnostic of spinal diseases. Methods: A retrospective analysis was performed on the cases of negative pressure biopsy conducted in our hospital from January 2015 to December 2017. A total of 52 cases, including 24 males and 28 females, were diagnosed as chest-back or lower back pain with vertebral body signal changes or destruction in imaging. It was difficult to make a clear diagnosis before surgery. Bilateral puncture is generally used for puncture of the affected vertebra under conventional local anesthesia in prone position. If the disease is obviously limited to one side of the vertebral body, unilateral puncture is selected. After establishing a channel, slowly screwing the trephine, stopping when reached the vertebral back edge in the lateral X-Ray, pull out slowly and check the presence of bone, after clearing the trephine, slowly screwing the trephine again, when observed the head of trephine is located in the front of the disease, stopped down. In the end of a trephine through 20 ml syringe infusion tee connection to the negative pressure and maintain, exit the trephine slowly gather syringe and tissue and blood clots in the trephine for pathologic examination. If the amount of tissue is small, it can be coaxial again into the trephine. When the trephine position on the lateral position slightly exceeds the lesion position, the negative pressure is established and the trephine is withdrawn. Specimens are collected and the working channel is withdrawn. The blood loss, operation time and pathological examination results were recorded and summarized. Results:One patient had much bleeding after the extraction of the trephine, and the bleeding stopped 3 minutes later after the gelatin sponge was filled with an internal bone cement filler. All the remaining patients successfully completed the surgery, with an average blood loss of less than 10ml and an average operation time of 15.82min. All the 48 cases had obvious bone tissue mass, 3 cases had soft tissue mass, and 1 case had no obvious macroscopic tissue mass. Results of histopathological examination: 34 cases of metastatic tumor, 5 cases of bone tuberculosis, 4 cases of myeloma, 3 cases of chronic inflammation, 1 case of lymphoma, 4 cases of bone tissue, 1 case of blood clot. The positive rate of puncture tissue was 98.08%, and the positive rate of puncture results was 90.38%. Conclusion: Using gear trephine under negative pressure can significantly improve the positive rate of vertebral body biopsy, which is a simple and effective diagnostic method for difficult spinal diseases