This study was aimed to work out a simple, applicable, sensitive and specific protocol for the identification of phosphoproteome. Isotope-labeling, two-dimensional electrophoresis, autoradiography and so on were used to establish a phosphoproteome map of mice neurons, and then chemiluminescence Western blotting was utilized to detect three phosphoproteins PI3Kr3, MEK1 and PKCalpha selectively. The results of comparison showed that the blots of PI3Kr3, MEK1 and PKCalpha on autoradiography map were almost identical with the blots of PI3Kr3, MEK1 and PKCalpha on chemiluminescence Western blotting maps. So this protocol based on isotope labeling and chemiluminescence Western blotting methods has proven to be sensitive and specific in the identification of phosphoproteome.
Animals ; Animals, Newborn ; Blotting, Western ; methods ; Brain ; cytology ; Cells, Cultured ; Electrophoresis, Gel, Two-Dimensional ; methods ; Luminescent Measurements ; methods ; Mice ; Neurons ; chemistry ; cytology ; Phosphoproteins ; analysis ; Phosphorus Radioisotopes ; Proteome ; analysis ; Sensitivity and Specificity

Objective To investigate the clinical value of 99Tcm-methoxyisobutylisonitrile (MIBI) double-phase imaging in differential diagnosis of bone lesions in tumor patients,which was indefinite in 99Tcm-methylene diphosphonate (MDP) bone imaging,and evaluate the correlation between 99Tcm-MIBI imaging and chemotherapy efficacy.Methods Fifty-two tumor patients (23 males,29 females,mean age 58 years) with bone isolated lesions(≤3) found by 99Tcm-MDP bone scintigraphy was enrolled from June 2014 to November 2015.Since the results of 99Tcm-MDP imaging were indefinite,99Tcm-MIBI double-phase (10 min and 30 min after injection of 99Tcm-MIBI) SPECT/CT imaging was then performed within 1 week.The final diagnosis was made according to results of more than two imaging modalities (CT,MRI,PET/CT) and/or follow-up (≥6 months).The diagnostic efficacy of 99Tcm-MIBI SPECT/CT imaging was calculated.The clinical dataof chemotherapy were also collected.Patients with bone metastasis were grouped as the treatment response.Retention index (RI) of metastatic lesions was calculated and compared between different groups using two-sample t test.The relationship between RI and chemotherapy efficacy was investigated by Spearman correlation analysis.Results A total of 12 benign lesions and 84 malignant lesions were eventually diagnosed.The sensitivity,specificity,positive predictive value (PPV),negative predictive value (NPV) and Youden index of 99Tcm-MIBI imaging were 96.43％(81/84),83.33％(10/12),97.59％(81/83),76.92％(10/13),and 0.80,respectively.The RI was statistically different between complete remission (CR)+partial remission (PR) group and stable disease (SD) + progressive disease (PD) group:-0.142± 0.036 vs-0.384± 0.067 (t =2.367,P＜0.05).The RI of 99Tcm-MIBI in bone metastases was positively correlated with the chemotherapy efficacy (rs =0.78,P＜0.01).Conclusions 99Tcm-MIBI imaging is helpful in differential diagnosis of bone lesions with indefinite diagnosis by 99Tcm-MDP bone imaging in tumor patients.99Tcm-MIBI RI of bone metastasis may predict the therapeutic response of chemotherapy efficacy.