ABSTRACT
Objective To explore circulating biomarkers for screening the invasiveness of non-functioning pituitary adenomas (NF-PAs). Methods The exosomal RNAs were extracted from serum of patients with invasive NF-PA (INF-PA) or noninvasive NF-PA (NNF-PA). Droplet digital PCR was adapted to detect the mRNA expression of candidate genes related to tumor progression or invasion, such as cyclin dependent kinase 6 (CDK6), ras homolog family member U (RHOU), and spire type actin nucleation factor 2 (SPIRE2). Student's -test was used to analyze the statistical difference in the mRNA expression of candidate genes between the two groups. Receiver operating characteristic (ROC) curve was used to establish a model for predicting the invasiveness of NF-PAs. The accuracy, sensitivity, specificity and precision of the model were then obtained to evaluate the diagnostic performance. Results CDK6 (0.2600±0.0912 . 0.1789±0.0628, =3.431, =0.0013) and RHOU mRNA expressions (0.2696±0.1118 . 0.1788±0.0857, =2.946, =0.0052) were upregulated in INF-PAs patients' serum exosomes as compared to NNF-PAs. For CDK6, the area under the ROC curve (AUC) was 0.772 (95% : 0.600-0.943, =0.005), the accuracy, sensitivity, specificity and precision were 77.27%, 83.33%, 75.00% and 55.56% to predict the invasiveness of NF-PAs. For RHOU, the AUC was 0.757 (95% : 0.599-0.915, =0.007), the accuracy, sensitivity, specificity and precision were 72.73%, 83.33%, 68.75% and 50.00%. In addition, the mRNA levels of CDK6 and RHOU in serum exosomes were significantly positively correlated (=0.935, <0.001). After combination of the cut-off scores of the two genes, the accuracy, sensitivity, specificity and precision were 81.82%, 83.33%, 81.25% and 62.50%. Conclusions CDK6 and RHOU mRNA in serum exosomes can be used as markers for predicting invasiveness of NF-PAs. Combination of the two genes performs better in distinguishing INF-PAs from NNF-PAs. These results indicate CDK6 and RHOU play important roles in the invasiveness of NF-PAs, and the established diagnostic method is valuable for directing the clinical screening and postoperative treatment.
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the protective effect of Exosomes from human adipose-derived mesenchymal stem cells (hAMSCs) in neural injury induced by glutamate and its possible mechanism.</p><p><b>METHODS</b>Characteristics of Exosomes from hAMSCs were identified by electron microscopy and Western blot analysis. Cytokines that might play a major role in the protective effect were tested by enzyme-linked immunosorbent assay (ELISA). The protective action of Exosome and its possible signaling pathway were researched by the in vitro neural injury induced by glutamate, including control group (without Glu), Glu group (dealing with Glu), Glu+Exo group (dealing with Glu +100 ng/ml Exo), Glu+Exo+Akt group (dealing with Glu+100 ng/ml Exo+10 μmol/L Akt), Glu+Exo+Erk group (dealing with 100 ng/ml Glu+100 ng/ml Exo+10 μmol/L Erk), and Glu+Exo+TrkB group (dealing with Glu+100 ng/ml Exo +10 μmol/L TrkB).</p><p><b>RESULTS</b>Exosomes from hAMSCs had similar sizes to those isolated from other kinds of cells, and expressed the characteristic proteins such as CD63, CD81, HSP70, and HSP90. Cytokines that had neurotrophic effects on Exosomes were mainly insulin-like growth factor and hepatocyte growth factor, with the concentration being 9336.49±258.63 and 58,645.50±16,014.62, respectively; brain derived neurotrophic factor, nerve growth factor,and vascular endothelial growth factor had lower levels, with the concentration being 1928.25±385.47, 1136.94±5.99, and 33.34±9.43, respectively. MTS assay showed that the PC12 cell survival rates were 0.842±0.047, 0.306±0.024, 0.566±0.026, 0.461±0.016, 0.497±0.003, and 0.515±0.034 in the control group, Glu group, Glu+Exo group, Glu+Exo+Akt group, Glu+Exo+Erk group, and Glu+Exo+TrkB group; obviously, it was significantly lower in Glu group than in control group (P=0.02), significantly higher in Glu+Exo group than in Glu group (P=0.01), and significantly lower in Glu+Exo+Akt group than in Glu+Exo group (P=0.01).</p><p><b>CONCLUSION</b>Exosomes secreted from hAMSCs have protective effect against neuron damage induced by glutamate, which may be mediated through activating the PI3/K-Akt signalling pathway.</p>
Subject(s)
Animals , Humans , Rats , Central Nervous System , Wounds and Injuries , Exosomes , Glutamic Acid , Mesenchymal Stem Cells , PC12 Cells , Vascular Endothelial Growth Factor AABSTRACT
The early diagnosis and treatment of pituitary carcinoma is difficult. The diagnosis is often delayed, and the confirmation of a diagnosis requires the presence of distant subarachnoid,brain or systemic metastasis from the primary pituitary tumor in the sella and also needs the evidences of pathology and imaging of the primary pituitary carcinoma and metastases. Treatment of pituitary carcinoma includes surgery, radiation therapy ,hormone therapy, chemotherapy, and molecularly targeted therapy; however, these methods are mainly palliative and can not prolong the survival. The prognosis remains poor. Efforts should be made to develop more effective diagnosis and treatment options.
Subject(s)
Humans , Diagnostic Imaging , Pituitary Neoplasms , PrognosisABSTRACT
Objectives: To improve the procedures for establishment of focal cerebral ischemic rat models with the intraluminal suture method and to elevate the survival rate of the models. Methods: Sixty male SD rats (weight, 250-270 g) were randomly allocated into 3 groups (n =20 in each group). Control group used the conventional surgical method, the pterygopalatine arteries were exposed and ligated during the procedure, and the rats had free access to food after the procedure; gavage group used the same method as the control group, but postoperative gavage was performed; protective group did not ligate the pterygopalatine arteries, but the carotid arteries and vagus nerves were carefully separated. Trying to avoid stretching the vagus nerves during the procedure and postoperative gavage was performed. The body weight of the rats in each group was weighed continuously before and after modeling, and the neurological deficit scores were measured; the survival rate of rats in each group was calculated at day 3, 14, and 28, respectively after modeling; the brain infarct volume in each group was calculated at day 28 after modeling. Results: Circled digit oneThe mean time of model production was 19.5 min in the protective group, and it was shorter than that in the control and gavage groups. The differences were statistically significant (P < 0.05); Circled digit twoAt day 3 after modeling, the survival rate of the rats in the protective group was higher than that in the control and gavage groups. The differences were statistically significant (P < 0.05); Circled digit threeAfter modeling, the body weight of the rats in the 3 groups were all decreased. The body weight of the rats in each group began to increase from day 7 after modeling. The body weight of the rats in the protective group was all higher than that in the control and gavage groups at day 7, 14, 21, and 28 after the procedure. The differences were statistically significant (P < 0.05); Circled digit fourthere were no significant differences comparing the neurological deficit scores and cerebral infarction volume among the 3 groups. Conclusion: The improved procedures could shorten the time of model production, regain body weight quickly, improve the survival rate effectively after establishment of the cerebral ischemic rat models.