[Objective]AIM To explore the differentiation of neural stem cells from rat embryonic spinalcord in the serum free condition culture medium of OECs.[Method]OECs were acquired by dissection ofolfactory nerve ensheathing and cultured for 20 hours,then suspending cells were put intoselective culture medium and cultured in the fibernectin coated plate.The activity of OECs was detected by MTT method on different periods and the best situation was selected out and serum-free cultured.Exudates of OECs was added into neural stem cells of the third generation.The differentiation of neural stem cells were observed under inverted microscope.The neural stemcells cultured were identified by immumofluorescence method.[Result]The activity of OECs was high on the 9~(th) day and the 12~(th) day.The serum free condition culture medium of OECs induced 53% neuralstem cells to differentiate into neuron-like cells and 42% neural stem cells into astrocytes cells.[Conclusion]The activity of OECs on different periods is different.The serum free condition culture medium of OECs can obviously induced neural stem cells to differentiate into mature neuronecells.

BACKGROUND: Microglia play an important role in immune surveillance in their quiescent state, but the role of the activated microglia is under discussion. OBJECTIVE: To analyze the mechanism of activated microglia in acute cerebral infarction. METHODS: Totally 96 male Kunming mice were selected and randomly divided into four groups, including transplantation, placebo, blank control and sham operation groups. Permanent occlusion of the middle cerebral artery was performed using suture method in the mice of the transplantation, placebo and blank control groups, followed by injection of microglia suspension via subclavian vein, medium containing the same volume of microglia, and nothing, respectively, at 12 hours after modeling. In the meanwhile, the same amount of microglia suspension was injected into the mice of the sham operation group. The Zea-longa scale and brain-derived neurotrophic factor expression at 12, 24 and 72 hours after modeling, the volume of cerebral infarction and the number of nerve cells positive for microtubule-associated protein-2 at 72 hours after modeling were detected. RESULTS AND CONCLUSION: The Zea-longa scale score was 0 point in the sham operation group, which was significantly lower than that in the other three groups at each time point after modeling (P < 0.01). The Zea-longa scores in the transplantation group were significantly lower than those in the placebo and blank control groups at 24 and 72 hours after transplantation (P < 0.01). The positive expression rate of brain-derived neurotrophic factor in the transplantation group was significantly higher than that in the other three groups after transplantation (P < 0.01). The sham group showed no infarction, while the size of cerebral infarction in the transplantation group was significantly lower than that in the placebo and blank control groups (P < 0.01), and the microtubule-associated protein-2 positive rate was significantly higher than that in the placebo and blank control groups (P < 0.01). These results manifest that the activated microglia can improve the survival rate of nerve cells, promote the recovery of cerebral nerve function and reduce the size of cerebral infarction.

Objective To summarize the clinical features of 22 probands diagnosed with congenital muscular dystrophy (CMD),and to provide genetic counseling and prenatal diagnosis for 23 fetuses of these pedigrees.Methods Data of 22 CMD patients who were treated in the Pediatric Department of Peking University First Hospital during October 2006 to March 2016 were analyzed.Informed written consents for participation in this study were obtained from the parents or guardians.Prenatal diagnosis was performed using DNA samples extracted from fetal villus cells of 12 cases at 11-13 gestational weeks and amniotic fluid of 11 cases at 18-22 gestational weeks.Direct DNA sequencing by polymerase chain reaction (PCR) and multiplex ligation-dependent probe amplification (MLPA) were used to detect CMD-related gene mutations.Linkage analysis of short tandem repeats (STRs) was used to identify maternal blood contamination and biological parents.Results Thirteen out of the 22 probands with CMD were diagnosed with congenital muscular dystrophy type 1 A (MDC1A),and all of them carried compound heterozygous mutations in LAMA2 gene.Prenatal diagnosis of 13 fetuses from these pedigrees found that four fetuses were wild-type,seven were heterozygotes and two carried the same mutations as their proband.Three probands with LMNA-related congenital muscular dystrophy (L-CMD) carried de novo mutations in LMNA gene.In these pedigrees,two fetuses were wild-type and one whose mother was mosaicism carried the same mutations as the proband.One proband with Ullrich congenital muscular dystrophy carried compound heterozygous mutations in COL6A2 gene and the fetus of the same pedigree was wild-type.Five probands were diagnosed with α-dystroglycanopathies.And among them,two cases of muscle-eye-brain disease (MEB) carried compound heterozygous mutations in POMGnT1 gene and the fetuses of the two peidgrees were heterozygotes;one case of congenital muscular dystrophy type 1C (MDC1C) had compound heterozygous mutations in FKRP gene and the fetus carried the same mutations;one patient diagnosed with POMGnT1-related congenital muscular dystrophy with mental retardation (CMD-MR) carried compound heterozygous mutations in POMGnT1 gene,and the fetus was positive for the same mutations;one proband with POMT1-related CMD-MR was positive for compound heterozygous mutations in POMT1 gene and the results of prenatal diagnosis for two fetuses of this pedigree showed that the first fetus had the same mutations as the proband,while the second was heterozygote.Conclusions No effective therapeutic method is available for CMD.Therefore,accurate genetic counseling and prenatal diagnosis are necessary to prevent CMD child from birth.

Objective:To evaluate the relationship between the tea drinking habit and postoperative delirium (POD) in elderly patients.Methods:Two hundred and ninety-two patients, aged 65-85 yr, weighing 50-80 kg, of American Society of Anesthesiologists physical status Ⅰ-Ⅲ, undergoing elective knee/hip arthroplasty under spinal-epidural anesthesia in our hospital, were enrolled in this study.The patient′s cognitive function was assessed using Mini-Mental State Examination at 1 day before operation.Peripheral venous blood samples were collected before anesthesia, and the concentrations of caffeine and tea polyphenols in plasma were measured by enzyme-linked immunosorbent assay.In the anesthesia recovery room after operation and at 1, 3 and 7 days after operation (or before discharge), neuropsychological tests were performed, and the Delirium Rating Scale was used to recognize POD developed.The patients were divided into POD group (P group) and non-POD group (NP group) according to whether POD occurred after operation.Logistic regression analysis was used to analyze the variables of which P values were less than 0.05. Results:There was no significant difference in age, American Society of Anesthesiologists physical status, concentrations of caffeine and tea polyphenols in plasma between P group and NP group ( P<0.05). The results of logistic regression analysis showed that age was an independent risk factor for POD, and concentrations of caffeine and tea polyphenols in plasma and tea drinking habits were protective factors for reducing the occurrence of POD in elderly patients. Conclusion:Tea drinking habit is a protective factor for reducing the occurrence of POD in elderly patients.

Objective:To evaluate the relationship between cholinergic biomarkers and postoperative delirium (POD) in elderly patients.Methods:The patients, aged 65-85 yr, weighing 50-80 kg, of American Society of Anesthesiologists physical status Ⅰ-Ⅲ, underwent total knee/hip arthroplasty under combined spinal-epidural block in our hospital from July 2018 to September 2019, were collected.The baseline clinical data of patients were collected, and cubital venous blood samples 5 ml were collected before anesthesia to detect plasma concentrations of choline acetyltransferase (ChAT), acetylcholinesterase (AChE), butyrylcholinesterase (BuChE), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6). The neuropsychological testing was performed on 1 day before operation, following admission to the recovery room after surgery, and on 1, 3 and 7 days (or before discharge) after surgery.The patient′s cognitive function was assessed using Mini-Mental State Examination (MMSE) before surgery.Confusion Assessment Method and Memorial Delirium Assessment Scale were used to evaluate the occurrence of postoperative delirium (POD) after surgery.The patients were divided into POD group (P group) and non-POD group (NP group) according to whether POD occurred.Logistic regression was used to analyze the related risk factors for POD.Results:There were 349 cases in NP group and 57 cases in P group, and the incidence of POD was 14.0%.Compared with NP group, the age of patients, preoperative coexisting underlying diseases (≥3 types), plasma ChAT, TNF-α and IL-6 concentrations were increased, and plasma AChE and BuChE concentrations were decreased in P group ( P<0.05). The results of multivariate logistic regression analysis showed that changes in plasma AChE, BuChE, and ChAT concentrations and older age were independent risk factors for POD ( P<0.05). Conclusion:The development of POD is related to the preoperative changes in plasma AChE, BuChE and ChAT concentrations in elderly patients.

Objective:To evaluate the effect of short-chain fatty acids on microglial synapse engulfment in aged rats with postoperative cognitive dysfunction (POCD).Methods:Forty-eight healthy male Sprague-Dawley rats, aged 18 months, weighing 520-650 g, were divided into 4 groups ( n=12 each) using a random number table method: control group (group C), short-chain fatty acids group (group S), POCD group (group P), and POCD+ short-chain fatty acids group (group PS). Rats received short-chain fatty acids (sodium propionate 25.9 mmol/L, sodium butyrate 40 mmol/L and sodium acetate 67.5 mmol/L) in the free drinking water for 28 days in S and PS groups. On day 29, anesthesia was induced with 4%-5% sevoflurane and maintained with 3% sevoflurane, and the tibial fracture internal fixation was performed to prepare a rat model of POCD in P group and PS group. Morris water maze test was performed at day 7 after surgery. The escape latency, times of crossing the original platform, mean swimming speed and time spent in the original platform quadrant were recorded. The rats were sacrificed at the end of Morris water maze test, and the brains were collected to analyze the number and density of dendritic spines in the hippocampal CA1 region (by Golgi staining) and to determine the expression of postsynaptic density 95 (PSD95) and complement 1q (C1q) in the hippocampal CA1 region (by immunofluorescence). Results:Compared with group C, the times of crossing the original platform were significantly decreased, the time spent in the original platform quadrant was shortened, the escape latency was prolonged, the number and density of dendritic spines and the number of intersection points between dendrites and concentric circles were decreased, the expression of PSD95 was down-regulated, and the expression of C1q was up-regulated in P and PS groups ( P<0.05). Compared with group P, the times of crossing the original platform were significantly increased, the time spent in the original platform quadrant was prolonged, the escape latency was shortened, the number and density of dendritic spines and the number of intersection points between dendrites and concentric circles were increased, the expression of PSD-95 was up-regulated, and the expression of C1q was down-regulated in group PS ( P<0.05). Conclusions:The mechanism by which short-chain fatty acids attenuates POCD is related to decreased microglial engulfment of synapses in aged rats.

Objective: To investigate the expression of integrin α5 in cervical cancer, and to explore its relationship with clinicopathological characteristics of patients with cervical cancer. Methods: Immunohistochemistry was used to detect the expression of integrin α 5 in cervical cancer tissues of 60 cases and normal cervical paraffin-embeded tissues of 20 cases of benign uterine lesions undergoing hysterectomy from Qingdao Municipal Hospital between January 2014 and December 2017. Real-time quantitative polymerase chain reaction (RT-PCR) was used to detect the mRNA expression level of integrin α5 in 20 fresh cervical cancer tissues and 20 normal cervical tissues collected from benign cervical lesions in Qingdao Municipal Hospital between January 2018 and July 2018. The relationship between the expression of integrin α5 and the clinicopathological characteristics of patients with cervical cancer was analyzed. Results: The positive expression rate of integrin α5 protein in cervical cancer and normal cervical tissues was 63.3% (38/60), 35.0% (7/20), respectively, and the difference was statistically significant (χ ² = 4.893, P < 0.05). The expression of integrin α5 mRNA in cervical cancer was 1.6±0.4 times as high as that in normal cervical tissues (t = 5.529, P < 0.01). The positive expression of integrin α5 protein was associated with lymph node metastasis in cervical cancer patients (Z = -2.636, P = 0.008). Conclusion The high expression of integrin α5 is related to lymph node metastasis of cervical cancer, and integrin α5 may be a new potential target for treatment of cervical cancer.

Polycomb group (PcG) ring finger protein 6 (PCGF6), though known as a member of the transcription-repressing complexes, PcG, also has activation function in regulating pluripotency gene expression. However, the mechanism underlying the activation function of PCGF6 is poorly understood. Here, we found that PCGF6 co-localizes to gene activation regions along with pluripotency factors such as OCT4. In addition, PCGF6 was recruited to a subset of the super-enhancer (SE) regions upstream of cell cycle-associated genes by OCT4, and increased their expression. By combining with promoter capture Hi-C data, we found that PCGF6 activates cell cycle genes by regulating SE-promoter interactions via 3D chromatin. Our findings highlight a novel mechanism of PcG protein in regulating pluripotency, and provide a research basis for the therapeutic application of pluripotent stem cells.