PFKP deubiquitination and stabilization by USP5 activate aerobic glycolysis to promote triple-negative breast cancer progression.

BACKGROUND: Triple-negative breast cancer (TNBC) remains the most challenging subtype of breast cancer and lacks definite treatment targets. Aerobic glycolysis is a hallmark of metabolic reprogramming that contributes to cancer progression. PFKP is a rate-limiting enzyme involved in aerobic glycolysis, which is overexpressed in various types of cancers. However, the underlying mechanisms and roles of the posttranslational modification of PFKP in TNBC remain unknown. METHODS: To explore whether PFKP protein has a potential role in the progression of TNBC, protein levels of PFKP in TNBC and normal breast tissues were examined by CPTAC database analysis, immunohistochemistry staining (IHC), and western blotting assay. Further CCK-8 assay, colony formation assay, EDU incorporation assay, and tumor xenograft experiments were used to detect the effect of PFKP on TNBC progression. To clarify the role of the USP5-PFKP pathway in TNBC progression, ubiquitin assay, co-immunoprecipitation (Co-IP), mass spectrometry-based protein identification, western blotting assay, immunofluorescence microscopy, in vitro binding assay, and glycolysis assay were conducted. RESULTS: Herein, we showed that PFKP protein was highly expressed in TNBC, which was associated with TNBC progression and poor prognosis of patients. In addition, we demonstrated that PFKP depletion significantly inhibited the TNBC progression in vitro and in vivo. Importantly, we identified that PFKP was a bona fide target of deubiquitinase USP5, and the USP5-mediated deubiquitination and stabilization of PFKP were essential for cancer cell aerobic glycolysis and TNBC progression. Moreover, we found a strong positive correlation between the expression of USP5 and PFKP in TNBC samples. Notably, the high expression of USP5 and PFKP was significantly correlated with poor clinical outcomes. CONCLUSIONS: Our study established the USP5-PFKP axis as an important regulatory mechanism of TNBC progression and provided a rationale for future therapeutic interventions in the treatment of TNBC.

Alamandine injected into the paraventricular nucleus increases blood pressure and sympathetic activation in spontaneously hypertensive rats.

Alamandine is a newly discovered new component of the renin-angiotensin (Ang) system (RAS) that has been shown to exert vasoactive effects in some areas of the nervous system. The present study investigated whether administration of alamandine to the hypothalamic paraventricular nucleus (PVN) modulates blood pressure and sympathetic activity. Mean arterial pressure (MAP) and renal sympathetic nerve activity (RSNA) were recorded in anaesthetized rats. PVN microinjection of alamandine increased MAP and RSNA both in Wistar-Kyoto (WKY) rats and in spontaneously hypertensive rats (SHRs), but to a greater extent in SHRs. Moreover, these effects were blocked by pretreatment with alamandine receptor Mas-related G-protein-coupled receptor, member D (MrgD) antagonist D-Pro7-Ang-(1-7), adenylyl cyclase (AC) inhibitor SQ22536, and protein kinase A (PKA) inhibitor rp-adenosine-3',5'-cyclic monophosphorothionate (Rp-cAMP). Treatment with D-Pro7-Ang-(1-7), SQ22536, or Rp-cAMP alone in PVN decreased MAP and RSNA in the SHRs. Conversely cAMP alone increased MAP and RSNA, and pretreatment with cAMP enhanced alamandine's effects. These results indicate that microinjection of alamandine into the PVN increases blood pressure and sympathetic outflow via MrgD and the cAMP-PKA pathway.

[Case-control study on minimally invasive percutaneous new plate osteosynthesis applied in proximal humerus fractures in elder patients].

OBJECTIVE: To evaluate the clinical significance of minimally invasive percutaneous new plate osteosynthesis (MIPPO) applied in three and four-part fractures of proximal humerus in elder patients. METHODS: From June 2008 to December 2010, 98 old patients with three and four-part fractures of proximal humerus were treated with locking plate osteosynthesis. There were 43 males and 55 females with an average age of (71.3 +/- 6.2) years ranging from 65 to 84. Among them,47 cases were treated with minimally invasive percutaneous new plate osteosynthesis (MIPPO) through antrolateral transdeoltoid approach, and meanwhile 51 cases were treated with open reduction and internal fixation (ORIF) through a traditional deltopectoral approach. The operation time,blood loss,wound complications and hospital stay were recorded. The functional outcomes of the shoulder were assessed according to Constant score. RESULTS: Ninety-six patients were followed up for 13 to 42 months (averaged 18.1 months),and 2 patients died during the follow-up period. The mean blood loss of the MIPPO group was lower than that of the ORIF group by 125 ml (P < 0.05). There was no difference in the operation time between two groups. But the MIPPO group had a slightly shorter hospital stay than the ORIF group (P < 0.05). All the fractures united clinically and radiologically. The difference of the mean healing time between two groups was not significant. The mean Constant score at final follow-up of two groups had no statistic difference. But the rate of complication of the MIPPO group was lower than that of the ORIF group (P < 0.05). Only 1 case of the MIPPO group had axillary nerve paralysis post-operatively, but 4 cases had slack internal fixation and 2 cases had avascular necrosis of the humeral head in the ORIF group. CONCLUSION: The technique of percutaneous insertion combined new locking plate internal fixation has the advantages of less blood loss, rapid recovery, less vascular damage, lower risk of complications and so on. It can effectively treat the proximal humerus three-and four-part fractures, especially to elder patients with osteoporosis.