BACKGROUND:Different types of nerve regulatory factors and glial cells have been reported to exert different roles in the differentiation and maturation of neural stem cells, but culturing neural stem cells in large animals is relatively rarely reported. OBJECTIVE:To explore the way for culturing goat neural stem cells and to detect the outcome after in vitro differentiation. METHODS:The neural stem cellwas separated and cultured from the newborn goat cerebral cortex and the anti-nestin immunocytochemical staining was performed for cellidentification. At the same time, anti-S100 active Schwann cells were gotten from the sciatic nerve. Then in vitro differentiation was preformed and the outcome was detected by the immunocytochemical stain of anti-glial fibril ary acidic protein, anti-microtubule-associated protein 2 and anti-S100. cells without primary antibodies served as controls. Gray values were calculated and compared. RESULTS AND CONCLUSION:The Schwann cells were cultured successful y, which were active to the anti-nestin immunocytochemical staining and anti-S100 staining. After differentiation, the products were active to anti-glial fibril ary acidic protein and anti-microtubule-associated protein 2 immunocytochemical stain, but Abstract BACKGROUND:Different types of nerve regulatory factors and glial cells have been reported to exert different roles in the differentiation and maturation of neural stem cells, but culturing neural stem cells in large animals is relatively rarely reported. OBJECTIVE:To explore the way for culturing goat neural stem cells and to detect the outcome after in vitro differentiation. METHODS:The neural stem cellwas separated and cultured from the newborn goat cerebral cortex and the anti-nestin immunocytochemical staining was performed for cellidentification. At the same time, anti-S100 active Schwann cells were gotten from the sciatic nerve. Then in vitro differentiation was preformed and the outcome was detected by the immunocytochemical stain of anti-glial fibril ary acidic protein, anti-microtubule-associated protein 2 and anti-S100. cells without primary antibodies served as controls. Gray values were calculated and compared. RESULTS AND CONCLUSION:The Schwann cells were cultured successful y, which were active to the anti-nestin immunocytochemical staining and anti-S100 staining. After differentiation, the products were active to anti-glial fibril ary acidic protein and anti-microtubule-associated protein 2 immunocytochemical stain, but negative to the anti-S100. And significant difference was found in gray values. The goat neural stem cells and Schwann cells were successful y cultured and identified. After the differentiation, the astrocytes and neurons were detected, but the Schwann cells were not found.

Objective:To evaluate the role of hippocampal REV-ERBα in postoperative cognitive dysfunction in rats.Methods:Thirty-two SPF healthy male Sprague-Dawley rats, aged 12-14 weeks, weighing 360-380 g, were divided into 4 groups ( n=8 each) using a random number table method: control group (group C), surgery group (group S), surgery + dimethyl sulfoxide (DMSO) group (group SD), and surgery + SR9009 group (group SS). Exploratory laparotomy was performed under sevoflurane anesthesia in S, SD and SS groups.Normal saline containing 0.1% DMSO was injected into hippocampal CA1 area at 1 h before laparotomy, with 2 μl on each side in group SD, and REV-ERBα agonist SR9009 (in normal saline containing 0.1% DMSO) was injected into hippocampal CA1 area at 1 h before laparotomy, with 2 μl on each side in group S+ SR9009.Morris water maze test was performed at 1 and 3 days after operation.Rats were sacrificed at 1 h after the end of Morris water maze test on day 3 after surgery, and the hippocampal tissues were obtained for determination of the expression of REV-ERBα, Brain and Muscle ARNT-Like 1 (BMAL1) protein, synaptophysin (SYN), postsynaptic density (PSD)-95 protein and N-methyl-D-aspartate receptor 2B subunit (GRIN2B) (by Western blot) and microscopic examination of the morphology of hippocampal neurons and Nissl bodies (by Nissl staining), and the viable neurons were counted. Results:Compared with group C, the percentage of time of staying at the target quadrant was significantly decreased, and the number of crossing platform was reduced on days 1 and 3 after exploratory laparotomy, the expression of REV-ERBα, BMAL1, PSD95, SYN and GRIN2B was down-regulated, and the number of viable neurons was decreased in group S and group SD ( P<0.05). Compared with group S and group SD, the percentage of time of staying at the target quadrant and the number of crossing platform were significantly increased on days 1 and 3 after exploratory laparotomy, the expression of REV-ERBα and PSD95 was up-regulated, the number of viable neurons was increased ( P<0.05), and no significant change was found in the expression of BMAL1, SYN and GRIN2B in group SS ( P>0.05). There was no significant difference in the indexes mentioned above between group S and group SD ( P>0.05). Conclusions:Activation of REV-ERBα can improve postoperative cognitive dysfunction, and the mechanism may be related to up-regulation of PSD95 expression in hippocampus and reduction of neuronal damage in rats.

Objective:To evaluate the role of hippocampal histone deacetylases (HDACs) in perioperative neurocognitive disorders (PND) and the relationship with postsynaptic dense protein 95 (PSD95) in rats.Methods:Sixty clean-grade healthy male Sprague-Dawley rats, aged 10-14 weeks, weighing 250-280 g, were divided into 3 groups ( n=20 each) using a random number table method: control group (group C), surgery group (group S) and HDAC inhibitor MS-275 group (group MS-275). Exploratory laparotomy was performed under 3% sevoflurane anesthesia in group S. MS-275 10 mg/kg was intraperitoneally injected at 0.5 h before exploratory laparotomy in group MS-275.Morris water maze tests were performed on 1 day before surgery and 1, 3 and 7 days after surgery.Ten rats were sacrificed on 1 day after surgery, and hippocampal tissues were obtained for determination of the expression of HDAC1-3 and PSD95 protein and mRNA by Western blot and real-time polymerase chain reaction, respectively.The density of hippocampal neurons was determined by the Nissl staining. Results:Compared with group C, the postoperative escape latency was significantly prolonged, the number of crossing the original platform was decreased, the density of hippocampal neurons was decreased, the expression of HDAC2 protein and mRNA was up-regulated, and the expression of PSD95 protein and mRNA was down-regulated in group S ( P<0.05 or 0.01). Compared with group S, the postoperative escape latency was significantly shortened, the number of crossing the original platform was increased, the density of hippocampal neurons was increased, the expression of HDAC2 protein and mRNA was down-regulated, and the expression of PSD95 protein and mRNA was up-regulated in group MS-275 ( P<0.05 or 0.01). There was no significant difference in the expression of HDAC1 and HDAC3 protein and mRNA among the three groups ( P>0.05). Conclusion:HDAC2 is involved in the pathophysiological mechanism of PND by down-regulating the expression of PSD95 in rats.

Objective:To investigate the protective effect of REV-ERBα agonist SR9009 on hippocampal working memory in rats with acute rapid eye movement (REM) sleep deprivation after exploratory laparotomy and its possible mechanism.Methods:Ninety SD rats were randomly divided into control group, sleep deprivation group, exploratory laparotomy group, sleep deprivation+exploratory laparotomy group, and sleep deprivation+exploratory laparotomy+SR9009 group ( n=18). Rats in the sleep deprivation group, exploratory laparotomy group, and sleep deprivation+exploratory laparotomy group were given REM sleep deprivation for 96 h or (and) exploratory laparotomy, respectively. Rats in the sleep deprivation+exploratory laparotomy+SR9009 group accepted exploratory laparotomy after REM sleep deprivation for 96 h, and accepted intraperitoneal injection of 100 mg/kg SR9009 daily from the day after surgery to the 6 th d of surgery. The reversall escape latency of rats was recorded by contrapuntal space exploration training one-5 d after surgery. On the 5 th d of surgery, reversal space exploration experiment was conducted to record the number of times of rats crossing the original platform. Western blotting was used to detect the protein expressions of REV-ERBα and BMAL1 in the hippocampus of rats. The levels of interleukin (IL)-1β and IL-6 in the hippocampus of rats were detected by enzyme-linked immunosorbent assay. Immunofluorescent staining was used to detect the expressions of neuronal nucleoprotein (NeuN) and glial fibrillary acidic protein (GFAP). Results:(1) The escape latency in the sleep deprivation group, exploratory laparotomy group, and sleep deprivation+exploratory laparotomy group was significantly longer than that in the control group on the first, 2 nd, 3 rd, 4 th, 5 th d of surgery ( P<0.05); while the escape latency in the sleep deprivation group and sleep deprivation+exploratory laparotomy group was significantly longer than that in the exploratory laparotomy group ( P<0.05); on the 2 nd, 3 rd, 4 th, 5 th d of surgery, the reversal escape latency in the sleep deprivation+exploratory laparotomy+SR9009 group was statistically shorter than that in the sleep deprivation+exploratory laparotomy group ( P<0.05). The number of times of rats crossing the original platform in the sleep deprivation group, exploratory laparotomy group, and sleep deprivation+exploratory laparotomy group was significantly smaller than that of the control group ( P<0.05); that of rats in the sleep deprivation+exploratory laparotomy group was significantly smaller than that of the exploratory laparotomy group, and that of rats in the sleep deprivation+exploratory laparotomy+SR9009 group was significantly larger than that of the sleep deprivation+exploratory laparotomy group ( P<0.05). (2) As compared with the control group, the exploratory laparotomy group, sleep deprivation group and sleep deprivation+exploratory laparotomy group had significantly decreased expressions of REV-ERBα and BMAL1, and statistically increased IL-1β and IL-6 levels in the hippocampal tissues ( P<0.05); as compared with the sleep deprivation+exploratory laparotomy group, the sleep deprivation+exploratory laparotomy+SR9009 group had significantly increased expressions of REV-ERBα and BMAL1, and statistically decreased IL-1β and IL-6 levels ( P<0.05). (3) As compared with the control group, the exploratory laparotomy group, sleep deprivation group and sleep deprivation+exploratory laparotomy group had decreased amount of neurons in the hippocampal CA3 area and increased amount of activated astrocytes; as compared with the sleep deprivation+exploratory laparotomy group, the sleep deprivation+exploratory laparotomy+SR9009 group had increased amount of neurons in the hippocampal CA3 area and decreased amount of activated astrocytes. Conclusion:Acute REM sleep deprivation can lead to work memory impairment in rats accepted exploratory laparotomy, which might be associated with neuroinflammation and REV-ERBα/BMAL1 pathway, and SR9009 could alleviate the damage.

At present the parkinsonian rigidity assessment depends on subjective judgment of neurologists according to their experience. This study presents a parkinsonian rigidity quantification system based on the electromechanical driving device and mechanical impedance measurement method. The quantification system applies the electromechanical driving device to perform the rigidity clinical assessment tasks (flexion-extension movements) in Parkinson's disease (PD) patients, which captures their motion and biomechanical information synchronously. Qualified rigidity features were obtained through statistical analysis method such as least-squares parameter estimation. By comparing the judgments from both the parkinsonian rigidity quantification system and neurologists, correlation analysis was performed to find the optimal quantitative feature. Clinical experiments showed that the mechanical impedance has the best correlation (Pearson correlation coefficient = 0.872, < 0.001) with the clinical unified Parkinson's disease rating scale (UPDRS) rigidity score. Results confirmed that this measurement system is capable of quantifying parkinsonian rigidity with advantages of simple operation and effective assessment. In addition, the mechanical impedance can be adopted to help doctors to diagnose and monitor parkinsonian rigidity objectively and accurately.