Objective To explore the best ratio of platelet-rich plasma (PRP) to activator and the synergistic action of thrombin and the growth factors in PRP gel on proliferation of human marrow-derived mesenchymal stem cells (hBMSCs). Methods The activator was made up of 1000 U bovine thrombin and 1 mL 10% calcium chloride solution. PRP was mixed with the activator at the ratios of 1:1, 5:1, 10:1, 20:1, 40:1 respectively in groups A, B, C, D and E. A quantitative sandwich enzyme-linked immunosorbent assay (ELISA) was used for examining the amounts of transforming growth factor-β1 (TGF-β1) and platelet derived growth factnr-AB (PDGF-AB) in PRP gel in each group after incubation for 0,1,8,24, and 120 hours. MTT assay was used to evaluate the effect of PRP gel in each group on hBMSCs proliferation. Results When PRP was mixed with thrombin, concentrations of PDGF-AB and TGF-β1 in PRP gel increased immediately and reached the peak value in 1 hour. The PRP gel in groups B, C, and D contained the highest amounts of PDGF-AB and TGF-β1, and accelerated hBMSCs growth re-markably. The cells proliferation in group A was inhibited. Conclusions The effect of PRP on the hBMSCs proliferation depends on the concentrations of PDGF-AB and TGF-β1 in PRP. Thrombin may influ-ence the hBMSCs proliferation by regulating concentrations of growth factors in PRP to certain extent, but too high a level of thrombin may inhibit hBMSCs growth.

Objective To explore the feasibility of labeling in vitro rabbit osteoblasts with Qtracker and the features of Qtracker-labeled rabbit osteoblasts. Methods A healthy male rabbit, 3 months old, weighing 2 kg, was used in this study. After bone marrow was aspirated, bone marrow stromal cells (BMSCs) were isolated and cultured using the adherence method in vitro. The third passage of BMSCs was induced into osteablasts before incubation with Qtracker at concentrations of 1, 2, 4, 8, 16, 32 nmol/10~6 cells (Groups A, B, C, D, E, F respectively). Cells not labeled by Qtracker served as negative control (Group G). The following parameters were measured: induction, differentiation and determination of rabbit osteoblasts; the optimal mass concentration of Qtracker labeling by fluorescence microscopy and flow cytometry; the cell sur-vival rates at various concentrations of Qtraeker labeling by trypan-blue exclusion; Qtracker-labeled cell pro-liferation by MTr. Results The primary and the passage rabbit BMSCs were chiefly of fusiform shape. Rabbit BMSCs differentiated into osteoblasts following induction. The osteoblasts cytoplasm showed green fluorescence under fluorescence microscopy after being labeled by Qtracker. The mean labeling rate increased with the increased concentration of Qtracker, reaching up to (93.58±2.08) % after incubation at 8 nmol/ 10~6 cells by fluorescence microscopy, and (95.24±1.31) % by flow cytometry. There were no significant differences between Groups D, E, F(P>0.05), but significant differences were found between Groups A, B, C and Groups D, E, F (P<0.05). The labeling rate for Group G was 0. The cell survival rates were all above 96% (P>0.05) . No significant differences were found in the cell proliferation among various con-centrations (P>0.05). Conclusions Qtraeker can be used as a labeling marker for rabbit osteoblasts. When the concentration is at 8 nmol/10~6 cells, optimal labeling effect can be achieved. Rabbit osteoblasts labeled with Qtracker are of high efficiency and safety.

Objective T o observe the changes of cystathionine β-synthase (C B S ) expression in the cere-bral cortex after brain contusion at different tim es. Methods A n experim ental m odel of traum atic brain injury (T B I) in m ice w as established by an im proved w eight-drop device. T hen W estern blotting and im m unohistochem ical exam ination w ere used to detect the C B S expression in cerebral cortex around in-jury at different tim e points (1 h, 6 h, 12 h, 1 d, 2 d, 3 d, 7 d). Results T he results of W estern blotting revealed that the expression level of C B S w as dow n-regulated and reached its low est level at the 3rd days after injury, and then restored to norm al level after 7 days. T he results of im m unohistochem istry show ed that C B S w as present in the norm al brain cortex. C B S expression gradually decreased at the 3rd days after injury, and then restored to norm al level after 7 days. Conclusion C B S has the potential to be a reference index for tim e estim ation after brain contusion in forensic practice.

Objective To explore the dynamic changes of glutamate in the cortex of cynomolgus monkeys during cerebral ischemia.Methods Proximal M1 segment of middle cerebral artery (MCA) was occluded for 1 h in 3 young cynomolgus monkeys (7.3 ± 1.5 years old) to induce cerebral ischemia.Magnetic resonance imaging and neurologic deficit scoring were used to evaluate the ischemia and observe the manifestations,respectively.Fast Analytical Sensing Technology (FAST) was applied to record the content of cortex glutamate in the same site of ipsilateral primary motor cortex in the periods of pre-,during,and post-occlusion,and at 1 and 2 weeks after surgery.Results Compared with pre-occlusion,the content of glutamate was increased significantly in the process of occluding in the MCA M1 (P =0.003);No significant difference was observed in the content during occluding and post-occlusion (P--0.877).The content in the first week was decreased obviously as compared with post-occlusion (P--0.004),but it showed no statistical difference with that in the second week (P =0.085).Conclusion Cerebral ischemia may potentially accelerate the extra-cellular glutamate release in the cortex,but reperfusion may ameliorate or balance off the glutamate release.

Objective To analyze the risk factors of hemorrhage after thrombolysis for mild cerebral infarction and the curative effect of low-dose alteplase.Methods 375 patients with mild cerebral infarction whose onset time was less than 4.5 hours from January 2013 to May 2017 in our hospital were selected as the research objects.Those undergoing low-dose alteplase thrombolysis were included in the thrombolysis group (n =195) while those without thrombolysis were included in the non-thrombolysis group (n =180).The therapeutic effects were analyzed.Patients with intracerebral hemorrhage in the thrombolysis group at 14 days after thrombolysis were included in the observation group while those without hemorrhage were included in the control group.Related data were compared between the two groups.The single factors and independent risk factors of hemorrhage after thrombolysis in patients with mild cerebral infarction were analyzed.Results National Institute of Health Stroke Scale (NIHSS) scores of the thrombolysis group at 1 week,1 month and 3 months after treatment were significantly lower than those of the non-thrombolysis group (P ＜0.05).Serum neuron specific enolase (NSE) and C-reactive protein (CRP) levels in the thrombolysis group were lower than those in the non-thrombolysis group after treatment (P ＜ 0.05).The prognosis of the thrombolysis group was better than that of the non-thrombolysis group after treatment (P ＜ 0.05).In this study,30 patients undergoing thrombolysis had intracerebral hemorrhage within 14 days after thrombolysis,and the incidence of intracranial hemorrhage was 15.38％ (30/195).Univariate and multivariate logistic regression analysis showed that age,history of heart failure,door to needle time (DNT),systolic blood pressure (SBP) in 12 hours after thrombolysis and leukoaraiosis change were risk factors for hemorrhage in patients with mild cerebral infarction after thrombolysis (P ＜ 0.05).Conclusions Using low-dose alteplase for treating patients with mild cerebral infarction can achieve good thrombolytic effects.Age,history of heart failure,DNT,SBP in 12 hours after thrombolysis and leukoaraiosis change are influencing factors of intracranial hemorrhage,which should be paid attention to.

Antiepileptic drugs (AEDs) are the main treatments of epilepsy, but their use is usually limited by adverse reactions, among them psychiatric and behavioral ones including aggressive behavior. Levetiracetam , perampanel, and topiramate , which have diverse mechanisms of action, have been associated with aggressive behavior. 5-HT and γ-aminobutyric acid (GABA), especially glutamate [via α-amino-3-hydroxy-5-methyl-4-isoxazole-propionate receptor (AMPA) receptor] seem to play key roles. Other mechanisms involve hormones, epigenetics and alternative psychosis. Neurological and/or mental health disorders tend to increase aggressive behavior sensitivity. The adverse reactions caused by AEDs are getting more and more attention. Clinicians may identify and manage this side effect as early as possible, if the risk factors and underlying mechanisms of aggressive behavior induced by antiepileptic we find. This article will explain the possible mechanisms.