Raddeanin A Protects the BRB Through Inhibiting Inflammation and Apoptosis in the Retina of Alzheimer's Disease.

Neuroinflammation and endothelial cell apoptosis are prominent features of blood-brain barrier (BBB) disruption, which have been described in Alzheimer's disease (AD) and can predict cognitive decline. Recent reports revealed vascular ß-amyloid (Aß) deposits, Muller cell degeneration and microglial dysfunction in the retina of AD patients. However, there has been no in-depth research on the roles of inflammation, retinal endothelial cell apoptosis, and blood-retinal barrier (BRB) damage in AD retinopathy. We found that Raddeanin A (RDA) could improve pathological and cognitive deficits in a mouse model of Alzheimer's disease by targeting ß-amyloidosis, However, the effects of RDA on AD retinal function require further study. To clarify whether RDA inhibits inflammation and apoptosis and thus improves BRB function in AD-related retinopathy. In vitro we used Aß-treated HRECs and MIO-M1 cells, and in vivo we used 3×Tg-AD mice to investigate the effect of RDA on BRB in AD-related retinopathy. We found that RDA could improve BRB function in AD-related retinopathy by inhibiting NLRP3-mediated inflammation and suppressing Wnt/ß-catenin pathway-mediated apoptosis, which is expected to improve the pathological changes in AD-related retinopathy and the quality of life of AD patients.

Design and synthesis of VEGFR-2 inhibitors based on oleanolic acid moiety.

In this study, twenty-four oleanolic acid (OA) derivatives were rationally designed based on molecule docking studies and their VEGFR-2 inhibitory activities were tested by Homogeneous time-resolved fluorescence (HTRF) method in vitro. All of the synthesized compounds were identified as new compounds, and the structures of these compounds were determined by 1H-NMR and ESI-MS. In the screening for VEGFR-2 inhibitors, compounds I6 and I7 exhibited excellent inhibitory effect. The results indicated that insertion of phenylurea group with a linker at position C-28 of OA can increase the activity against VEGFR-2 significantly. [Formula: see text].

Dexmedetomidine Protects Neural Stem Cells from Ketamine-Induced Injury.

BACKGROUND/AIMS: Ketamine inhibits the proliferation of neural stem cells (NSCs) and disturbs normal neurogenesis. Dexmedetomidine provides neuroprotection against volatile anesthetic-induced neuroapoptosis and cognitive impairment in the developing brain. Whether it may protect NSCs from ketamine-induced injury remains unknown. In this study, we investigated the protective effects of dexmedetomidine on ketamine-exposed NSCs and explored the mechanisms potentially involved. METHODS: Primary NSC cultures were characterized using immunofluorescence. Cell viability was determined using a Cell Counting Kit 8 assay. Proliferation and apoptosis were assessed with BrdU incorporation and TUNEL assays, respectively. Protein levels of cleaved caspase-3, phosphorylated protein kinase B (p-Akt), and glycogen synthase kinase-3ß (p-GSK-3ß) were quantified using western blotting. RESULTS: Ket-amine significantly decreased NSC viability and proliferation and increased their apoptosis. Dexmedetomidine increased NSC proliferation and decreased their apoptosis in a dose-dependent manner. Furthermore, dexmedetomidine pretreatment notably augmented the viability and proliferation of ketamine-exposed NSCs and reduced their apoptosis. Moreover, dexmedetomidine lessened caspase-3 activation and increased p-Akt and p-GSK-3ß levels in NSCs exposed to ketamine. The protective effects of dexmedetomidine on ketamine-exposed NSCs could be partly reversed by the PI3K inhibitor LY294002. CONCLUSIONS: Collectively, these findings indicate that dexmedetomidine may protect NSCs from ketamine-induced injury via the PI3K/Akt/GSK-3ß signaling pathway.

Simvastatin Attenuates Neurogenetic Damage and Improves Neurocongnitive Deficits Induced by Isoflurane in Neonatal Rats.

BACKGROUND/AIMS: Isoflurane inhibited neurogenesis and induced subsequent neurocognitive deficits in developing brain. Simvastatin exerts neuroprotection in a wide range of brain injury models. In the present study, we investigated whether simvastatin could attenuate neurogenetic inhibition and cognitive deficits induced by isoflurane exposure in neonatal rats. METHODS: Sprague-Dawley rats at postnatal day (PND) 7 and neural stem cells (NSCs) were treated with either gas mixture, isoflurane, or simvastatin 60 min prior to isoflurane exposure, respectively. The rats were decapitated at PND 8 and PND 10 for detection of neurogenesis in the subventricular zone (SVZ) and subgranular zone (SGZ) of the hippocampus by immunostaining. NSC proliferation, viability and apoptosis were assessed by immunohistochemistry, CCK-8 and TUNEL, respectively. The protein expressions of caspase-3, p-Akt and p-GSK-3ß both in vivo and vitro were assessed by western blotting. Cognitive functions were assessed by Morris Water Maze test and context fear conditioning test at the adult. RESULTS: Isoflurane exposure inhibited neurogenesis in the SVZ and SGZ, decreased NSC proliferation and viability, promoted NSC apoptosis and led to late cognitive deficits. Furthermore, isoflurane increased caspase-3 expression and decreased protein expressions of p-Akt and p-GSK-3ß both in vivo and in vitro. Pretreatment with simvastatin attenuated isoflurane-elicited changes in NSCs and cognitive function. Co-treatment with LY294002 reversed the effect of simvastatin on NSCs in vitro. CONCLUSION: We for the first time showed that simvastatin, by upregulating Akt/GSK-3ß signaling pathway, alleviated isoflurane-induced neurogenetic damage and neurocognitive deficits in developing rat brain.

Correlation Between Flow-Mediated Dilatation of the Brachial Artery and Serum Endothelial Biomarkers in the Evaluation of Acute Endothelial Injury After Cardiopulmonary Bypass.

OBJECTIVES: The purpose of this study was to investigate the correlation between flow-mediated dilatation of the brachial artery and serum endothelial biomarkers and to discuss the feasibility of sonographic evaluation of acute endothelial injury during cardiopulmonary bypass (CPB) surgery. METHODS: Sonography was applied to determine the percentage of change in the brachial artery size during flow-mediated dilatation. Meanwhile, the plasma concentrations of endothelial-derived biomarkers, such as endothelin 1, nitric oxide, and von Willebrand factor, were measured to monitor the changes in endothelial function. We analyzed the correlation between flow-mediated dilatation and biomarkers during the perioperative period of CPB in 27 patients. RESULTS: All of the biomarkers changed dramatically, especially during the CPB period. There was a negative correlation between flow-mediated dilatation and von Willebrand factor (P = .001; R = -0.31). CONCLUSIONS: A CPB event has a substantial impact on endothelial function, and sonographic assessment of the percentage of change in the brachial artery size during flow-mediated dilatation allows early detection of acute endothelial function injury in cardiac surgery.

Physio-psycho-social interaction mechanism in dyadic health of young and middle-aged stroke survivors and their spousal caregivers: a longitudinal observational study protocol.

INTRODUCTION: In recent years, stroke has become more common among young people. Stroke not only has a profound impact on patients' health but also incurs stress and health threats to their caregivers, especially spousal caregivers. Moreover, the health of stroke survivors and their caregivers is interdependent. To our knowledge, no study has explored dyadic health of young and middle-aged stroke survivors and their spousal caregivers from physiological, psychological and social perspectives. Therefore, this proposed study aims to explore the mechanism of how physiological, psychological and social factors affect dyadic health of young and middle-aged stroke survivors and their spousal caregivers. The findings of this study will provide implications for developing interventions to improve dyadic health of this growing population. METHODS AND ANALYSES: We will collect data from 57 dyads of young and middle-aged stroke survivors and their spousal caregivers during hospitalisation and at 1, 3, 6, 9 and 12 months after discharge. Questionnaires will be used to collect participants' demographic information, stress, depression, anxiety, benefit finding, social support, mutuality and quality of life. The following physiological reactions will be collected at baseline, including interleukin 6, tumour necrosis factor-alpha and salivary cortisol. ETHICS AND DISSEMINATION: The study was approved by the ethics review committee of life sciences of Zhengzhou University (No. ZZUIRB2020-53). Prior to being enrolled in the study, participants will be given full and detailed information about the possible risks involved, the informed consent process, confidentiality, the study procedure and secure data storage. Participants will be guaranteed that they can withdraw from the study at any time without providing a reason or leading to any consequences. Both oral and written informed consent will be obtained from all participants. The findings of this proposed study will be disseminated through peer-reviewed journals and academic conferences.

[Culture and identification of neural stem cells from mouse embryos].

OBJECTIVE: The purpose of this study was to culture and identify neural stem cells from mouse embryos in vitro using a modified method and provide a basis for further study of the biology of neural stem cells under hypoxia. METHODS: The cells were isolated mechanically from the front cortex of fetal Institute of Cancer Research (ICR) mice on embryonic day 14. They were passaged by mechanical dissociation and enzymatic digestion. The neurospheres were identified by immunofluorescent staining of nestin. Cell differentiation was induced by 1% fetal bovine serum and then the cells were identified by immunohistochemistry of ß-tubulin III and GFAP. RESULTS: The cells obtained from the front cortex of fetal ICR mice had the capacity of forming neurospheres which showed nestin immunoreactive positivity. After being induced by 1% fetal bovine serum, the cells were differentiated into ß-tubulin III-positive cells and GFAP-positive cells. CONCLUSIONS: Using mechanical dissociation of primary cells and mechanical dissociation with enzymatic digestion of primary cells, the NSCs from the front cortex of mouse embryos can be obtained.

Cytokine-induced killer cells carrying recombinant oncolytic adenovirus expressing p21Ras scFv inhibited liver cancer.

Background: Oncolytic adenovirus-mediated gene therapy is an emerging strategy for cancer treatment. However, oncolytic adenoviruses are mainly administered locally at tumor site. Intravenous administration of oncolytic adenovirus for cancer gene therapy is a problem that needs to be solved urgently. Methods: We constructed recombinant oncolytic adenovirus KGHV500 carrying anti-p21Ras scFv and employed CIK cells to deliver KGHV500. TUNEL, wound healing, MTT, and Transwell invasion assays were used to determine the anti-tumor efficacy of KGHV500 on liver cancer cells in vitro. Nude mouse xenograft model was used to examine the anti-tumor efficacy of CIK cells combined with KGHV500 in vivo. Furthermore, KGHV500 accumulation in different organs was detected to assess the safety. Results: KGHV500 inhibited the migration, proliferation, invasion, and induced the apoptosis of liver cancer cells. CIK cells carrying KGHV500 reached tumor site and exerted much better anti-tumor efficacy than CIK cells or KGHV500 alone in nude mouse xenograft model. Moreover, we detected KGHV500 and anti-p21Ras scFv in different organs of nude mice, with little effects on the organs. Conclusions: We develop a novel strategy for the treatment of Ras-driven liver cancer by combining CIK cells with oncolytic adenovirus expressing anti-p21Ras scFv. Intravenous injection of CIK cells carrying KGHV500 in vivo significantly inhibits tumor growth, has little effect on normal organs, and is relatively safe.

DUOX2 As a Potential Prognostic Marker which Promotes Cell Motility and Proliferation in Pancreatic Cancer.

DUOX2 has been reported to highly express in several types of cancers. However, the prognostic significance and the biological function of DUOX2 expression with pancreatic cancer (PC) still remain unclear. The present study is aimed at investigating whether DUOX2 could act as a novel biomarker of prognosis and evaluating its effect on PC cell progression. The mRNA and protein expression of DUOX2 in PC cells and tissues were assessed by quantitative real-time PCR (RT-qPCR) and immunohistochemistry. The effect of DUOX2 expression on PC cell motility and proliferation was evaluated in vitro. The correlation between DUOX2 mRNA expression and clinicopathological features and its prognostic significance were analyzed according to the Gene Expression Profiling Interactive Analysis (GEPIA) website based on The Cancer Genome Atlas (TCGA) and the GTEx databases combined with our clinical information. According to bioinformatics analysis, we forecasted the upstream transcription factors (TFs) and microRNA (miRNA) regulatory mechanism of DUOX2 in PC. The expression of DUOX2 at transcriptional and protein level was dramatically increased in PC specimens when compared to adjacent nontumor specimens. Functionally, DUOX2 knockdown inhibited cell motility and proliferation activities. Our clinical data revealed that the patients had better postoperative overall survival (OS) with lower expression of DUOX2, which is consistent with GEPIA data. Multivariate analysis revealed that high DUOX2 expression was considered as an independent prognostic indicator for OS (P = 0.031). Based on Cistrome database, the top 5 TFs of each positively and negatively association with DUOX2 were predicted. hsa-miR-5193 and hsa-miR-1343-3p targeting DUOX2 were forecasted from TargetScan, miRDB, and DIANA-TarBase databases, which were negatively correlated with OS (P = 0.043 and P = 0.0088, respectively) and DUOX2 expression (P = 0.0093 and P = 0.0032, respectively) in PC from TCGA data. These findings suggest that DUOX2 acts as a promising predictive biomarker and an oncogene in PC, which could be a therapeutic target for PC.

A new learning paradigm for random vector functional-link network: RVFL.

In school, a teacher plays an important role in various classroom teaching patterns. Likewise to this human learning activity, the learning using privileged information (LUPI) paradigm provides additional information generated by the teacher to 'teach' learning models during the training stage. Therefore, this novel learning paradigm is a typical Teacher-Student Interaction mechanism. This paper is the first to present a random vector functional link (RVFL) network based on the LUPI paradigm, called RVFL+. The novel RVFL+ incorporates the LUPI paradigm that can leverage additional source of information into the RVFL, which offers an alternative way to train the RVFL. Rather than simply combining two existing approaches, the newly-derived RVFL+ fills the gap between classical randomized neural networks and the newfashioned LUPI paradigm. Moreover, the proposed RVFL+ can perform in conjunction with the kernel trick for highly complicated nonlinear feature learning, termed KRVFL+. Furthermore, the statistical property of the proposed RVFL+ is investigated, and the authors present a sharp and high-quality generalization error bound based on the Rademacher complexity. Competitive experimental results on 14 real-world datasets illustrate the great effectiveness and efficiency of the novel RVFL+ and KRVFL+, which can achieve better generalization performance than state-of-the-art methods.

CUL1 Knockdown Attenuates the Adhesion, Invasion, and Migration of Triple-Negative Breast Cancer Cells via Inhibition of Epithelial-Mesenchymal Transition.

Cullin-1 (CUL1) is an important factor for tumor growth and a potential therapeutic target for breast cancer therapy, but the molecular mechanism in triple-negative breast cancer (TNBC) is unknown. In the present study, CUL1 shRNA was transfected into BT549 and MDA-MB-231 breast cancer cells. Cell morphology, adhesion, invasion, and migration assays were carried out in the CUL1 knockdown cells. Additionally, protein expression levels of epithelial-mesenchymal transition (EMT)-related factors, Akt phosphorylation at S473 (pAkt), glycogen synthase kinase-3ß phosphorylation at ser9 (pGSK3ß), cytoplasmic and nuclear ß-catenin, and epidermal growth factor receptor phosphorylation at Tyr1068 (pEGFR) were detected by Western blot analysis. CUL1 knockdown significantly suppressed the adhesion, invasion and migration capabilities of the cells, and decreased the expression of Snail1/2, ZEB1/2, Twist1/2, Vimentin, and increased the expression of Cytokeratin 18 (CK18). Moreover, CUL1 knockdown significantly downregulated the phosphorylated levels of Akt, GSK3ß, and EGFR, inhibiting the translocation of ß-catenin from the cytoplasm to the nucleus. The results indicate that CUL1 knockdown prohibited the metastasis behaviors of breast cancer cells through downregulation (dephosphorylation) of the EMT signaling pathways of EGFR and Akt/GSK3ß/ß-catenin in breast cancer. These results strongly suggested that reinforcement of the EMT might be a key for CUL1 to accelerate TNBC metastasis.

Effects of banana powder (Musa acuminata Colla) on the composition of human fecal microbiota and metabolic output using in vitro fermentation.

Bananas are rich in indigestible carbohydrates and are considered potential whole-fruit prebiotics. To investigate banana-induced changes in the composition of the human gut microbiota and the production of short chain fatty acids (SCFAs), ripe banana (Musa acuminata Colla, Degrees Brix: 22.6 ± 0.2° Bé), from Hainan, China, was powdered and fermented in vitro for 24 hr with the feces of six Chinese donors. The degradation of banana polysaccharides was observed in all six fecal samples. During in vitro fecal fermentation, banana polysaccharides were gradually degraded up to approximately 80%. The production of SCFAs was also measured. The addition of banana powder increased the concentrations of acetate, propionate, and butyrate, with the production of acetate being higher than that of propionate and butyrate. Changes in the human gut microbiota were assessed using high-throughput sequencing of the 16S ribosomal RNA (rRNA) gene. The results indicated that banana powder significantly altered bacterial diversity, increasing the relative abundance of Bacteroides, while maintaining the proportion of Bifidobacterium in the feces. At the same time, banana powder also increased the proportion of Lactobacillus; however, a significant difference was not observed. In summary, banana powder can be utilized by specific bacteria in human intestines, providing data support for the study of the effects of banana powder on the human intestinal health. PRACTICAL APPLICATION: In this study, in vitro batch fermentation was used to evaluate the effect of banana powder on the human intestinal microbial community, and the metabolized products of banana powder were determined. Our study showed that banana powder improved the human intestinal microbial flora and promoted the growth of Bifidobacterium and Bacteroides and could produce beneficial SCFAs (acetate, propionate, and butyrate). This study provided a theoretical basis for the use of banana powder as a potential prebiotic in production applications and our daily diet.

[Effect of prenatal heroin exposure on p-ERK1/2 expression in the prefrontal lobe cortex, hippocampus and nucleus accumbens in mice].

OBJECTIVE: To study the expression of phosphorylated-ERK1/2(p-ERK1/2)MAPK in the prefrontal lobe cortex (PFC), hippocampus (HP) and nucleus accumbens (Acb) in mice exposed to heroin in the uterus, and elucidate whether ERK MAPK signal transduction pathway participates in neurobehavioral teratogenicity induced by maternal heroin abuse. METHODS: Animal model was established by subcutaneous administration of diacetylmorphine (10 mg/kg.d) to pregnant BALB/c mice on embryonic days 9-18, and their offspring were assigned to heroin and normal saline groups according to the maternal treatment. P-ERK1/2 expression in the PFC, HP and Acb were detected by RT-PCR and Western blot. RESULTS: The heroin group had body weights similar to the normal saline group after birth. There were no significant differences in the p-ERK1/2 expression in the PFC, HP and Acb between the two groups. CONCLUSIONS: Prenatal exposure to 10 mg/kg heroin altered neither the body weight nor the general development in mice. The ERK1/2 MAPK signal pathway might not be involved in the neurobehavioral teratogenicity induced by prenatal heroin exposure.

RAS mutations in acute myeloid leukaemia patients: A review and meta-analysis.

RAS oncogene mutations frequently occur in acute myeloid leukaemia (AML), but the prognostic significance of RAS mutations in AML is inconclusive. We searched the databases of PubMed, Web of Science, EMBASE, and Cochrane from 1990 to 2018. In this study, 24 eligible studies were included, and the meta-analysis was conducted with the Comprehensive Meta-Analysis Version 2 software program. The row hazard ratio (HR) was adjusted and re-evaluated when publication bias existed after detecting all the heterogeneities. A combined analysis showed that RAS mutations were not associated with a poor prognosis in general AML patients (HR: 0.96, 95% CI: 0.78-1.19, p = 0.70). To further verify the results, a subgroup analysis was conducted. Interestingly, in the analysis of age bracket, children with RAS mutations had an unfavourable survival (HR: 1.35, 95% CI: 1.05-1.75, p = 0.02) of AML, but the adults did not (HR: 0.87, 95% CI: 0.70-1.09, p = 0.21). Further analysis of the subgroup of children indicated that patients with NRAS mutations had an adverse prognosis (HR: 1.55, 95% CI: 1.13-2.12, p = 0.007), but not those with KRAS mutations (HR: 1.51, 95% CI: 0.34-6.73, p = 0.59). In conclusion, this study revealed that RAS mutations did not influence the over survival for adults with AML. However, NRAS mutations may be a key prognostic marker related with poor survival for children with AML.

Changes of serum lipopolysaccharide, inflammatory factors, and cecal microbiota in obese rats with type 2 diabetes induced by Roux-en-Y gastric bypass.

OBJECTIVES: Previous studies have shown that Roux-en-Y gastric bypass (RYGB) leads to rapid regression of obesity and type 2 diabetes (T2D). However, the underlying mechanism remains unclear. This study aimed to investigate the effect of RYGB on serum lipopolysaccharide (LPS), interleukin (IL)-1, IL-6, tumor necrosis factor alpha (TNF-α), and cecal microbiota in obese rats with T2D. METHODS: Obese Sprague-Dawley rats with T2D were randomly divided into RYGB diabetes operation (DO; n = 8), diabetes sham operation (DS; n = 8), and diabetic control (DC; n = 8) groups. Healthy Sprague-Dawley rats were grouped as normal control (NC; n = 8). Fasting plasma glucose and body weight were measured. The levels of peripheral serum LPS, IL-1, IL-6, and TNF-α were measured by enzyme-linked immunosorbent assay. The rats were sacrificed 12 wk after operation. Subsequently, a superior mesenteric venous blood sample was taken to measure serum LPS levels by enzyme-linked immunosorbent assay. The cecal contents of the DO and DS groups were taken to extract metagenomic DNA per the genomic DNA standardization procedure. The V4 region of the 16 S rRNA was sequenced with the Illumina Hiseq sequencing platform to compare the structure and relative abundance of cecal microbiota between the DO and DS groups. RESULTS: Twelve weeks after operation in the DO group, fasting plasma glucose and body weight showed a significant decrease (P < 0.05). Moreover, the levels of peripheral serum LPS, IL-1, IL-6, and TNF-α were obviously decreased (P < 0.05). A change in the LPS level of superior mesenteric venous blood also revealed a dramatic decrease (P < 0.05). Additionally, RYGB resulted in a shift of cecal microbiota in obese rats with T2D. CONCLUSIONS: Hypoglycemic effects after RYGB may be associated with improved levels of LPS, IL-1, IL-6, and TNF-α. Changes in the structure of cecal microbiota may also play an important role.

A Novel AdaBoost Framework With Robust Threshold and Structural Optimization.

The AdaBoost algorithm is a popular ensemble method that combines several weak learners to boost generalization performance. However, conventional AdaBoost.RT algorithms suffer from the limitation that the threshold value must be manually specified rather than chosen through a self-adaptive mechanism, which cannot guarantee a result in an optimal model for general cases. In this paper, we present a generic AdaBoost framework with robust threshold mechanism and structural optimization on regression problems. The error statistics of each weak learner on one given problem dataset is utilized to automate the choice of the optimal cut-off threshold value. In addition, a special single-layer neural network is employed to provide a second opportunity to further adjust the structure and strength the adaption capability of the AdaBoost regression model. Moreover, to consolidate the theoretical foundation of AdaBoost algorithms, we are the first to conduct a rigorous and comprehensive theoretical analysis on the proposed approach. We prove that the general bound on the empirical error with a fraction of training examples is always within a limited soft margin, which indicates that our novel algorithm can avoid over-fitting. We further analyze the bounds on the generalization error directly under probably approximately correct learning. The extensive experimental verifications on the UCI benchmarks have demonstrated that the performance of the proposed method is superior to other state-of-the-art ensemble and single learning algorithms. Furthermore, a real-world indoor positioning application has also revealed that the proposed method has higher positioning accuracy and faster speed.

[In vivo labeling of cells in the ependyma/subventricular zone of adult rats].

OBJECTIVE: To identify the labeled cells in the ependyma/subventricular zone (SVZ) of normal adult rats by DiI injected into the lateral ventricle. METHODS: Fifty male Sprague-Dawley rats were divided randomly into five groups (10 per group). All of the rats were injected with 10 microL of 2 g/L fluorescence dye DiI into the right lateral ventricle. The five groups of rats were sacrificed at 6 h, 12 h, 24 h, 36 h or 48 h after injection respectively. Hoechst 33258 staining was used to identify nuclei and laser confocal microscopy was used to detect the DiI-labeled cells and to measure the thickness of the tissue with DiI fluorescence in the wall of the left lateral ventricle. RESULTS: After injection of the DiI into the right lateral ventricle, DiI- Hoechst 33258 double positive cells were found in the ependymal layer of the left lateral ventricular wall at 24 h and in the SVZ at 48 h as well. The thickness of the tissue with DiI fluorescence in the left ependyma/septal subventricular zone (SVZspt) and ependyma/postnatal equivalent of the ganglionic eminences (SVZge) remained unchanged at 12 h and 24 h after DiI injection. The thickness of the tissue with DiI fluorescence in the left ependyma/SVZge was significantly greater than that in the ependyma/SVZspt at all of the time points (P<0.05). CONCLUSION: The ependyma/SVZ cells can be labeled by Dil 24-48 h after injection (10 microL of 2 g/L) into the lateral ventricle.

[Effect of ligustrazine on cell proliferation in subventricular zone in rat brain with focal cerebral ischemia-reperfusion injury].

OBJECTIVE: To observe the effect of ligustrazine on cell proliferation in subventricular zone (SVZ) in rat brain with focal cerebral ischemia reperfusion injury. METHODS: Male SD rats were randomly divided into a normal group,a sham operation group,a ligustrazine treatment group, and a control group. The ligustrazine treatment group and the control group were further divided into 5 subgroups: 1d, 3d, 7d, 14d, and 21d reperfusion after 2h middle cerebral artery occlusion (MCAO). The focal cerebral ischemia-reperfusion model was made by MCAO. S phase cells were labelled with BrdU. Immunohistochemistry method was conducted to detect the BrdU positive cells. The total number of BrdU positive cells in the SVZ was measured. The expression of neuro nitric oxide synthase (nNOS) was detected with Western blot method. RESULTS: There was a significant increase of BrdU positive cells in SVZ of ligustrazine treatment in the 1d and 3d group compared with that of the control group (P<0.01). The total number of BrdU positive cells reached a peak in 7d group and declined afterwards. Cells proliferated also in SVZ on the contralateral side, and peaked at 7d. The nNOS expression of ligustrazine administration after the focal cerebral ischemia-reperfusion decreased at 1d and 3d after the reperfusion compared with that of the control group (P<0.05), and increased at 7d, but with no significant difference compared with that of the control group. CONCLUSION: Ligustrazine may promote the cell proliferation in SVZ of adult rats with ischemia-reperfusion injury by decreasing the nNOS expression.

Effects of ketamine-midazolam anesthesia on the expression of NMDA and AMPA receptor subunit in the peri-infarction of rat brain.

BACKGROUND: Activation of N-methyl-D-aspartate (NMDA) receptors and alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA) receptors play an important role in the neurons death induced by ischemia. The mitigating effect of intravenous anesthetics on ischemic neuron injury is related to their influence on NMDA receptors. This study was performed to investigate the effect of ketamine-midazolam anesthesia on the NMDA and AMPA receptor subunits expression in the peri-infarction of ischemic rat brain and explore its potential mechanism of neuroprotection. METHODS: Thirty Sprague Dawley (SD) rats were subjected to permanent middle cerebral artery occlusion under ketamine/atropine (100/0.05 mg/kg) or ketamine-midazolam/atropine (60/50/0.05 mg/kg) intraperitoneal anesthesia (n=15 each). Twenty-four hours after ischemia, five rats in each group were killed by injecting the above dosage of ketamine or ketamine-midazolam intraperitoneally and infarct size was measured. Twenty-four and 72 hours after ischemia, four rats in each group were killed by injecting the above dosage of ketamine or ketamine-midazolam intraperitoneally. After staining the brain tissue slices with toluidine blue, the survived neurons in the peri-infarction were observed. Also, the expression level of NMDA receptors 1 (NR1), NMDA receptors 2A (NR2A), NMDA receptors 2B (NR2B) and AMPA (GluR1 subunit) were determined by grayscale analysis in immunohistochemical stained slices. RESULTS: Compared with ketamine anesthesia, ketamine-midazolam anesthesia produced not only smaller infarct size [(24.1+/-4.6)% vs (38.4+/-4.2)%, P<0.05], but also higher neuron density (24 hours: 846+/-16 vs 756+/-24, P<0.05; 72 hours: 882+/-22 vs 785+/-18, P<0.05) and lower NR2A (24 hours: 123.0+/-4.9 vs 95.0+/-2.5, P<0.05; 72 hours: 77.8+/-4.1 vs 54.2+/-3.9, P<0.05) and NR2B (24 hours: 98.5+/-2.7 vs 76.3+/-2.4, P<0.05; 72 hours: 67.2 +/-7.5 vs 22.2+/-2.6, P<0.05) expression level in the peri-infarction following ischemia. CONCLUSION: The protective effects of ketamine-midazolam anesthesia on ischemic brain injury may related to decreasing NR2A and NR2B expression.

[A comparison study for differentiation between human fetal retinal progenitor cells and brain neural stem cells in vitro].

OBJECTIVE: Investigating the potential of differentiation of human fetal retinal progenitor cells (hRPCs) and brain neural stem cells (hBNSCs) in vitro. METHODS: hRPCs and hBNSCs were isolated from human fetuses (8-12 weeks of gestation) and cultured in serum-free DMEM/F12 culture medium with N2 supplement, epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) or culture medium with 10% fetal bovine serum (FBS) but without EGF and bFGF. Immunocytochemical and immunofluorescence studies were conducted for identification of neural stem cells, retinal progenitors or the subtypes of neurons, astrocytes, retinal ganglion cells and rod photoreceptors with the specific antibodies for Nestin, Pax6, Map2, GFAP, Thy-1 and Rhodopsin, respectively. RESULTS: Both hRPCs and hBNSCs could proliferate and differentiate in DMEM/F12 + N2 with or without 10% FBS and expressed specific markers of immature neuroepithelial cells, retinal progenitors, mature neurons, astrocytes, retinal ganglion cells and rod photoreceptors. hBNSCs easily attached, spread out longer neurites and to form a network when cultured with serum contained medium. hRPCs were more difficult to attach and had only short dendrites. CONCLUSIONS: Both hRPCs and hBNSCs can differentiate into retinal specific cell types in vitro. The adherent, migration and differential capacity of hRPCs and hBNSCs are different when these cells are induced by the serum-contained culture medium.