Objective To examine the role of p38MAPK in high glucose-induced apoptosis of osteoblast MC3T3-E1 cell line, and to investigate its effect on the expressions of apoptosis-related molecules including caspase3, bax, and bcl-2. Methods The lentiviral vector containing short hairpin RNA targeting p38MAPK was constructed. The cultured osteoblast MC3T3-E1 cell were divided into 5 groups:normal control group(A group), high glucose group(B group), p38MAPK-shRNA transfection group(C group), signal transduction inhibitor group(D group), and transfection with negative control siRNA group(E group). RT-PCR was used to determine the p38MAPK mRNA expression levels in MC3T3-E1 cells. Flow cytometry(FCM)was employed to detect the cell apoptotic percentage. The protein levels of apoptosis-related molecules p38MAPK, p-p38MAPK, caspase-3, bax, and bcl-2 were assayed by Western blot. Ultrastructural alternation of MC3T3-E1 cell was observed under transmission electron microscopy(TEM). Results The lentiviral vector containing short hairp in RNA targeting p38MAPK was successfully constructed and transfected into MC3T3-E1 cells. RT-PCR result suggested that the siRNA targeting p38MAPK could effectively reduce the p38MAPK mRNA expression level induced by high glucose in MC3T3-E1 cell line. FCM showed siRNA significantly decreased high glucose-induced apoptosis percentage of MC3T3-E1 cells(P＜0.01). Meanwhile, we also found the siRNA significantly attenuated the proteins levels of p38MAPK, p-p38MAPK, caspase-3, and gene bax induced by high glucose in MC3T3-E1 cells, whereas the protein level of gene bcl-2 was enhanced remarkably when compared with high glucose group and negative control siRNA group(P＜0.01, P＜0.05).Conclusion The iRNA targeting p38MAPK suppressed high glucose-induced MC3T3-E1 cell apoptosis via inhibiting the activation of p38MAPK signaling pathway, thereby reducing the expressions levels of p-p38MAPK, caspase-3 and gene bax, and up-regulating the level of gene bcl-2.

Objective:To investigate the characteristics and evolution of mild motor symptoms (MMS) in patients with prodromal Parkinson′s disease (pPD).Methods:Based on the pPD cohort screened by Parkinson′s Disease Prodromal Clinical Assessment Scale in Nanjing community from July 2018 to December 2020, the clinical data of 30 patients with pPD who completed the baseline assessment and were followed up for at least 1 year were analyzed. According to the Unified Parkinson Diease Rating Scale Ⅲ (UPDRS-Ⅲ) score, the patients were divided into MMS group (UPDRS-Ⅲ score>3) and non-MMS group (NMMS group, UPDRS-Ⅲ score≤3). The differences and evolution characteristics of clinical characteristics between the 2 groups were compared. Multivariate linear regression was used to analyze the risk factors of motor symptom progression in pPD patients.Results:Among the 30 patients with pPD, 7 of 23 patients in the MMS group were converted to PD at the end of follow-up, 1 of 7 patients in the NMMS group were converted to PD at the end of follow-up. The UPDRS-Ⅲ score [10.00 (7.00, 17.00)], Montreal Cognitive Assessment Scale (MoCA) score [25.50 (24.75, 28.00)] and the Hamilton Anxiety Scale (HAMA) score [9.00 (5.00, 13.00)] at the end of follow-up of pPD patients were significantly higher than those at baseline [7.00 (4.00, 12.00), 24.00 (22.75, 25.25) and 8.00 (2.00, 11.00)], and the differences were statistically significant ( Z=-3.505, P<0.001; Z=-2.956, P=0.003; Z=-2.427, P=0.015).Subgroup analysis showed that UPDRS-Ⅲ score [11.00 (7.00, 18.00)], MoCA score [25.00 (24.00, 27.00)] and HAMA score [ 9.00 (6.00, 15.00)] at the end of follow-up in the MMS group were higher than those at baseline [8.00 (6.00, 12.00), 24.00 (22.00, 25.00) and 9.00 (3.00, 11.00)], and the difference was statistically significant (Z=-2.768, P=0.006; Z=-2.457, P=0.014; Z=-2.250, P=0.024). The Non-Motor Symptoms Questionnaire score at the end of follow-up in the MMS group (8.96±5.20) was significantly lower than that in the baseline (11.04±4.41), and the difference was statistically significant ( t=2.441, P=0.023).There was no significant difference in Mini-Mental State Examination (MMSE), Hamilton Depression Scale (HAMD), Rapid Eyes Movement Sleep Behavior Disorder Questionnaire-Hong Kong (RBDQ-HK) and Sniffin′ sticks olfactory test score at the end of follow-up in the MMS group. Only UPDRS-Ⅲ score in the NMMS group was increased at the end of follow-up [7.00 (5.00, 8.00)] compared with the baseline [4.00 (1.00, 4.00)], and the difference was statistically significant ( Z=-2.375, P=0.018). There was no significant difference in MoCA, MMSE, HAMA, HAMD, RBDQ-HK, and Sniffin′ sticks olfactory test score between the NMMS group and the baseline at the end of follow-up. Conclusion:The clinical conversion rate of pPD patients with MMS is high,and screening of this population should be paid attention.