HEK293 or HeLa cells were transfected by NIRF and, or P53, whole cell extracts andimmunoprecipitates were subjected to SDS-PAGE followed by Western blotting. GST pull-down was carried outto identify the interactions between NIRF and P53. In vitro ubiquitination reaction was carried out to identify P53ubquitinate by NIRF. The results suggested that NIRF could interact with P53 in vivo and in vitro. The results alsoshowed that NIRF could ubiquitinate P53 in vivo and in vitro. The results indicated that NIRF would be a newnegative regulator of P53.

Objective:To investigate the extracellular signal-regulated kinase 5 (ERK5) and matrix metallo proteinase-9 (MMP-9) expres-sion levels in osteosarcoma tissues and their clinical significance. Methods:The ERK5 and MMP-9 expression levels in 71 specimens of osteosarcoma tissue and 40 specimens of normal bone tissue were detected by immunohistochemistry. The relationship between ERK5 and MMP-9 expression levels, their clinical characteristics, and prognosis of patients with osteosarcoma were analyzed. Results:The positive expression of ERK5 and MMP-9 in osteosarcoma tissues was 85.9%(61/71) and 74.65%(53/71), respectively, which were significantly higher than those in normal bone tissues at 12.5%(5/40) and 10.0%(4/40) (all P<0.05). The positive expression of ERK5 and MMP-9 was associated with Enneking stage and metastasis (all P<0.05). Kaplan-Meier analysis showed that the survival duration of patients with positive ERK5 and MMP-9 expression levels was shorter than those of the patients in the negative expression groups (all P<0.05). Univariate analysis of COX proportional hazards regression model revealed that tumor size, Enneking stage, metastasis, and positive ERK5 and MMP-9 expression levels are relevant to the overall survival of patients with osteosarcoma (all P<0.05). Multi-variate analysis of COX proportional hazards regression model confirmed that Enneking stage, metastasis, and positive ERK5 and MMP-9 expression levels can act as independent prognostic factors for osteosarcoma patients (all P<0.05). Conclusion:The ERK5 and MMP-9 expression levels are high in osteosarcoma tissues and are related to the clinical characteristics and prognosis of patients with osteo-sarcoma. Thus, ERK5 and MMP-9 expression levels may play important roles in osteosarcoma development and progression.

To summarize the experience of perinatal nursing for 27 days of delayed delivery of the second fetus of a twin pregnancy with placenta previa.Key nursing points include:the start of high risk warning during delayed delivery,and being alert to placenta previa hemorrhage;effective prevention of infection,reducing the incidence of maternal sepsis and chorioamnionitis;actively inhibiting the contractions to prolong gestational age and improving the perinatal survival rate of the second fetus;strengthening in utero fetal monitoring,identifying the early signs of fetal distress;targeted implementation of personalized psychological nursing,to ensure the mental health of patients.The rescue plan during delivery should be improved to ensure the safety of mother and child.Postpartum prevention and treatment of massive bleeding was actively conducted,as well as the prevention of puerperal infection.After 79 d of active treatment and intensive nursing care,the patient was treated and discharged from the hospital without any signs of infection or a massive bleeding.The second newborn,whose delivery was delayed,was discharged from the hospital 59 d after birth,and the follow up was continued for a month until the reach to the standard growth development parameters and the overall physical examination was good.