Objective To study the clinical efficacy and safety of hydroxyurea combined with thalidomide in the treatment of JAK2V617F positive myeloproliferative neoplasms (MPN).Methods A prospective analysis was conducted, involving 80 patients diagnosed with JAK2V617F positive myeloproliferative neoplasms via quantitative PCR at our hospital from January 2020 to January 2023. The patients were divided into an observation group (40 cases) and a control group (40 cases) based on the treatment method. The observation group received a combination of hydroxyurea and thalidomide, while the control group received conventional thalidomide treatment alone. The clinical efficacy, incidence of adverse reactions and bone marrow morphology were compared between the two groups, as well as the JAK2V617F mutation burden, the levels of platelet (PLT), white blood cell (WBC), hemoglobin (HGB) and hematocrit (HCT), and the myelofibrosis degrees of the two groups before treatment and 6 and 12 months after treatment.Results After treatment, the total effective rate of the observation group was significantly higher than that of the control group (P<0.05). At 6 and 12 months after treatment, the JAK2V617F mutation load, the levels of PLT and HGB, the MPN-10 score and MF grade in the two groups were significantly lower than those before treatment, and those in the observation group were significantly lower than those in the control group (P<0.05). However, there was no significant change in WBC and HCT levels (P>0.05). The incidence of abnormal proliferation of bone marrow cells in the observation group was significantly lower than that in the control group (P<0.05). No statistically significant differences were found in the incidence of adverse reactions between the two groups (P>0.05).Conclusion Hydroxyurea combined with thalidomide treatment is effective for JAK2V617F positive myeloproliferative neoplasm patients. It can significantly improve the clinical symptoms, reduce JAK2V617F mutation burden, and reverse myelofibrosis.