Curcumin prevents Alzheimer's disease progression by upregulating JMJD3.

The main purpose of this research was to explore the molecular mechanisms of Jumonji Domain-Containing Protein 3 (JMJD3) in Alzheimer's disease (AD) and to analyze its role in the anti-AD mechanism of curcumin (CUR). In the in vitro study of AD, JMJD3 overexpression promoted the trimethylation of histone H3 lysine 27 (H3K27me3), downregulated brain-derived neurotrophic factor (BDNF ), improved the abnormality of mitochondrial stress response (MSR) markers, Aß accumulation, increased cell proliferation and inhibited apoptosis. Upregulating BDNF also achieved above similar results. Knockout of JMJD3 could downregulate BDNF, upregulate the level of H3K27me3 methylation and inhibit MSR markers, while transfection of JMJD3 RNAi could counteract the upregulated effect of BDNF. Then, MSR activator could also improve AD. In addition, JMJD3 in AD in vitro models was obviously upregulated under CUR stimulation, and it triggered a series of reactions as mentioned above. In the in vivo study, the levels of JMJD3, the mRNA and protein levels of BDNF in the right brain tissues of AD mice were downregulated, the methylation of H3K27me3 increased, and the MSR markers (ClpP, HSP6, HSP-60, ATFS-1, etc.) were downregulated; the above indexes were improved in varying degrees with the intervention of CUR. Thus, we conclude that CUR can induce the upregulation of JMJD3 and improve BDNF expression by promoting the demethylation of H3K27me3, thereby maintaining the balance of MSR and thus, preventing AD development.

Effect of main family caregiver's anxiety and depression on mortality of patients with moderate-severe stroke.

Anxiety and depression are common mental illness in stroke caregivers, resulting in significant stress to the emotion health of caregivers. Caregivers' emotion can seriously affect the recovery rate of stroke patient, therefore, how to control and affect the caregivers' anxiety and depression is of great importance. Here three multiple centers observation and validation study were performed to screen out the risk factors for development of anxiety and depression in main family caregiver, and the effect of anxiety and depression of family caregivers on 6-month mortality of patients with moderate-severe stroke. The severity of the stroke, the duration of care time and the medical payment associated with increased risk of anxiety and depression. Anxiety and depression of main family caregivers are associated with increased risk 6-month mortality of patients with moderate-severe stroke. Therefore, the support provided to the family caregivers might have positive effect on prognosis of the patients with stroke.

Curcumin-Activated Mesenchymal Stem Cells Derived from Human Umbilical Cord and Their Effects on MPTP-Mouse Model of Parkinson's Disease: A New Biological Therapy for Parkinson's Disease.

BACKGROUND: The aim of this study was to investigate the effects of human umbilical cord mesenchymal stem cell activated by curcumin (hUC-MSCs-CUR) on Parkinson's disease (PD). hUC-MSCs can differentiate into many types of adult tissue cells including dopaminergic (DA) neurons. CUR could protect DA neurons from apoptosis induced by 6-hydroxydopamine (6-OHDA). Therefore, we used the hUC-MSCs activated by CUR for the treatment of PD in an animal model. METHODS: The hUC-MSCs-CUR was transplanted into the MPTP-induced PD mouse models via the tail vein. We found that hUC-MSCs-CUR significantly improved the motor ability, increased the tyrosine hydroxylase (TH), dopamine (DA), and Bcl-2 levels, and reduced nitric oxide synthase, Bax, and cleaved caspase 3 expression in PD mice. The supernatant of hUC-MSCs-CUR (CM-CUR) was used to stimulate the SH-SY5Y cellular model of PD; cell proliferation, differentiation, TH, and neuronal-specific marker microtubular-associated protein 2 (MAP2) expressions were examined. RESULTS: Our data showed that CM-CUR significantly promoted cell proliferation and gradually increased TH and MAP2 expression in SH-SY5Y PD cells. The beneficial effects could be associated with significant increase of rough endoplasmic reticulum in the hUC-MSCs-CUR, which secretes many cytokines and growth factors beneficial for PD treatment. CONCLUSIONS: Transplantation of hUC-MSCs-CUR could show promise for improving the motor recovery of PD.

Magnetic resonance imaging and risk factors for progression of lacunar infarct lesions in Chinese patients.

PURPOSE: The proportion of acute symptomatic lacunar infarction lesions that undergo cavitation and the factors influencing cavity formation are yet unclear, particularly in the Chinese population. Hence, we investigated changes in the diameter of acute lacunar infarction lesions and identified the risk factors for the progression of these lesions. METHODS: A total of 160 patients (mean age 66 years) with acute symptomatic lacunar infarction lesions underwent two magnetic resonance imaging (MRI) examinations: diffusion-weighted imaging (DWI) at onset (lesion diameter < 20 mm) and T2-weighted imaging/fluid-attenuated inversion recovery sequences at follow-up (median follow-up time 389 days). Lacunar infarction lesion progression was categorized as complete cavitation (lacune), partial cavitation, white matter lesion (WML), or disappearance of the lesion. The risk factors for cavity formation were evaluated. RESULTS: Upon follow-up MRI, lesions had changed to lacunes in 20 (12.5%) patients, partial cavitation in 23 (14.4%), WMLs in 97 (60.6%), and had disappeared in 20 (12.5%). Lacune formation was related to hypertension (P = 0.026); cavity (lacune and partial cavitation) formation was related to diabetes (P = 0.009) and diameter change (P = 0.015). CONCLUSIONS: Approximately a quarter of the acute symptomatic lacunar infarction lesions observed with follow-up MRI were cavitated. Hypertension was negatively associated with lacune formation; diabetes and diameter change were negatively associated with cavity formation.

Haplotype analysis on association between variants of interleukin 6 (IL-6) and late-onset Alzheimer's disease in a Chinese Han population.

OBJECTIVE: The aim of this study was to evaluate the influence of single nucleotide polymorphisms (SNPs) in interleukin-6 (IL-6) gene and the haplotype on late-onset Alzheimer's disease (LOAD). METHODS: A total of 896 participants were enrolled, including 446 LOAD patients and 450 controls. Total genomic DNA was extracted from the blood of participants and genotyping was then performed. Hardy-Weinberg equilibrium test was conducted in controls. Multivariate analysis was performed to determine the association between 4 SNPs (rs1800796, rs7802308, rs1800795 and rs13447446) in IL-6 gene and LOAD. Pairwise LD analysis was employed to identify the association between haplotype and LOAD. RESULTS: Multivariate logistic analysis showed that T allele of rs7802308 and G allele of rs1800796 were correlated with decreased risk of LOAD, adjusted ORs were 0.68 (95% CI: 0.50-0.91) and 0.71 (95% CI: 0.51-0.92), respectively. However, rs1800795 and rs13447446 were not associated with LOAD. Pairwise LD analysis among the 4 SNPs showed that only rs1800796 and rs1800795 in IL-6 gene was in heavy LD, the D' value was >0.75 (0.825). In all samples, the haplotype C-G was observed most frequently in two groups, with 55.79% and 49.46% in the LOAD patients and controls, respectively. The results also indicated that haplotype G-C was significantly associated with decreased LOAG risk. CONCLUSIONS: T allele of rs7802308 and G allele of rs1800796 were correlated with decreased risk of LOAD. The haplotype G-C were also correlated with decreased risk of LOAD.

MicroRNA-129-5p alleviates nerve injury and inflammatory response of Alzheimer's disease via downregulating SOX6.

There is growing evidence of the position of microRNAs (miRs) in Alzheimer's disease (AD), thus our objective was to discuss the impact of miR-129-5p regulating nerve injury and inflammatory response in AD rats by modulating SOX6 expression. The AD rat model was established by injecting Aß25-35 into the brain. The pathological changes, ultrastructure, number of neurons, cell degeneration and apoptosis of hippocampal tissue were observed in vivo. MiR-129-5p, SOX6, IL-1ß, TNF-α, Bcl-2 and Bax expression in serum and hippocampal tissues were detected by ELISA, RT-qPCR or western blot analysis. The successfully modeled hippocampal neuronal cells of AD were transfected with miR-129-5p mimic, SOX6-siRNA or their controls to figure out their roles in proliferation, apoptosis and inflammatory reaction in vitro. Low expression of SOX6 and high expression of miR-129-5p in vivo of rats would shorten the escape latent period and increase the times of crossing platforms, alleviate the pathological injury, inhibit neuronal apoptosis and reduce the inflammatory reaction. Up-regulation of miR-129-5p and down-regulation of SOX6 promoted proliferation, suppressed apoptosis and degraded the inflammatory reaction of neuronal cells in vitro. Up-regulation of SOX6 reversed the expression of miR-129-5p to reduce the damage and inflammatory response of the cell model of AD. Our study presents that up-regulation of miR-129-5p or down-regulation of SOX6 can reduce nerve injury and inflammatory response in rats with AD. Thus, miR-129-5p may be a potential candidate for the treatment of AD.

SPK1-transfected UCMSC has better therapeutic activity than UCMSC in the treatment of experimental autoimmune encephalomyelitis model of Multiple sclerosis.

Multiple Sclerosis (MS), is a chronic inflammatory autoimmune disorder of the central nervous system that leads to chronic demyelination with axonal damage and neuronal loss. Mesenchymal stem cells (MSCs) represent a promising therapeutic approach for MS. In the current study, we investigated the effects of MSCs derived from the human umbilical cord (UCMSC) transfected by sphingosine kinase 1 (SPK1) gene. All the results showed that transplantation of UCMSCs gene modified by SPK1 (UCMSC-SPK1) dramatically reduce the severity of neurological deficits of the experimental autoimmune encephalomyelitis (EAE) mice, paralleling by reductions in demyelination, axonal loss, and astrogliosis. UCMSC-SPK1 transplantation also could inhibit the development of natural killer (NK) responses in the spleen of EAE mice, and increase the ratio of CD4+ CD25+ FoxP3+ (Treg) T cells. Furthermore, we described that a shift in the cytokine response from Th1/Th17 to Th2 was an underlying mechanism that suppressed CNS autoimmunity. UCMSCs transfected by SPK1 gene potentially offer a novel mode for the treatment of MS, and the specific mechanism of SPK1 in treating MS/EAE.

Relationship of polymorphisms and haplotype in interleukin-16 and adiponectin gene with late-onset Alzheimer's disease risk.

AIMS: To investigate the impact of Interleukin-16 (IL- 16) and Adiponectin (ANP) gene single nucleotide polymorphisms (SNPs), gene- gene interactions and haplotype on late-onset Alzheimer's disease (LOAD) risk. METHODS: Hardy-Weinberg equilibrium (HWE), haplotype and pairwise linkage disequilibrium (LD) analysis were investigated by using SNPstats (available online at http://bioinfo.iconcologia.net/SNPstats). Generalized multifactor dimensionality reduction (GMDR) was used to examine interaction among 4 SNPs, odds ratio (OR) and 95% confident interval (95%CI) were calculated by logistic regression model. RESULTS: LOAD risk was significantly higher in carriers of rs266729- G allele than those with CC genotype (CG+ GG versus CC), OR (95%CI) =1.61 (1.26-1.96), and higher in carriers of rs1501299- T allele, OR (95%CI) = 1.62 (1.32-2.12), lower in carriers of rs4072111- T allele, adjusted OR (95%CI) =0.65 (0.44-0.93). We also found a significant gene- gene interaction between rs266729 and rs4072111. Participants with CG or GG of rs266729 and CC of rs4072111 genotype have the highest LOAD risk, OR (95%CI) = 2.62 (1.64 -3.58). Haplotype containing the rs266729- G and rs1501299- T alleles were associated with increased LOAD risk, OR (95%CI)= 1.83 (1.32- 2.43), and haplotype containing the rs1131445- C and rs4072111- T alleles were associated with decreased LOAD risk, OR (95%CI)= 0.53 (0.18- 0.95). CONCLUSIONS: We concluded that rs266729 and rs1501299 minor alleles were associated with increased LOAD risk, but rs4072111 minor allele was associated with decreased LOAD risk. We also found that interaction involving rs266729 and rs4072111, and haplotype combinations were associated with LOAD risk.

Protective Effect of Curcumin Against Oxidative Stress-Induced Injury in Rats with Parkinson's Disease Through the Wnt/ ß-Catenin Signaling Pathway.

BACKGROUND/AIMS: The study aimed to investigate the protective effect of curcumin against oxidative stress-induced injury of Parkinson's disease (PD) through the Wnt/ß-catenin signaling pathway in rats. METHODS: The successfully established PD rat models and normal healthy rats were randomly assigned into the 6-hydroxydopamine (6-OHDA), the curcumin (Cur) and the control groups. Immunohistochemistry was used to detect the positive expression of tyrosine hydroxylase (TH), dopamine transporter (DAT) and glial fibrillary acidic protein (GFAP). Deutocerebrum primary cells were extracted and classified into the control, 6-OHDA, Cur (5, 10, 15 µmol/L), Dickkopf-1 (DKK-1) and Cur + DKK-1 groups. MTT assays, adhesion tests and TUNEL staining were used to assess cell viability, adhesion and apoptosis, respectively. Western blotting and qRT-PCR were used to examine the protein and mRNA expressions of Wnt3a and ß-catenin and the c-myc and cyclinD1 mRNA expressions. RESULTS: TH and DAT expressions in the Cur group were elevated and GFAP was reduced compared with the 6-OHDA group. Curcumin enhanced viability, survival and adhesion and attenuated apoptosis of deutocerebrum primary cells by activating the Wnt/ß-catenin signaling pathway. Higher Wnt3a and ß-catenin mRNA and protein expressions and c-myc and cyclinD1 mRNA expressions, enhanced superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) contents, decreased malondialdehyde (MDA) content and elevated mitochondrial membrane potential (∆ψm) were found in the 10 and 15 µmol/L Cur groups compared with the 6-OHDA group. However, opposite tendencies were found in the Cur + DKK-1 group compared to the 10 µmol/L Cur group. CONCLUSION: This study suggests that curcumin could protect against oxidative stress-induced injury in PD rats via the Wnt/ß-catenin signaling pathway.

Gene- gene interaction between CYP2J2 and PPAR -γ gene on late-onset Alzheimer's disease in the eastern Chinese Han population.

AIMS: To investigate the impact of PPAR -γ and CYP2J2 gene single nucleotide polymorphisms (SNPs) and gene- gene interactions on late-onset Alzheimer's disease (LOAD) risk in Chinese Han population. METHODS: A total of 880 participants (514 males, 366 females), with a mean age of 81.7±15.9years old are selected, including 430 LOAD patients and 450 normal participants. Generalized multifactor dimensionality reduction (GMDR) is used to examine interaction among six SNPs, odds ratio (OR) and 95% confident interval (95%CI) are calculated by Logistic regression model. RESULTS: Logistic regression analysis showed that increased LOAD risks are associated with T allele of the rs1155002 polymorphism, adjusted OR (95%CI)=1.46 (1.12-1.90), and T allele of the rs890293 polymorphism, adjusted OR (95%CI)=1.65 (1.30-2.06), and G allele of the rs1805192 polymorphism, adjusted OR (95%CI)=1.70 (1.25-2.27). We also found a potential gene-gene interaction between rs890293 and rs1805192. Participants with GT or TT of rs890293 and CG or GG of rs1805192 genotype have the highest LOAD risk, compared to participants with GG of rs890293 and CC of rs1805192 genotype, OR (95%CI)=2.22 (1.63 -2.85), after covariates adjustment. CONCLUSIONS: rs1155002, rs890293 and rs1805192 polymorphism are associated with increased LOAD risk. Participants with GT or TT of rs890293 and CG or GG of rs1805192 genotype have the highest LOAD risk.

Effects of caffeic acid on learning deficits in a model of Alzheimer's disease.

Caffeic acid is a type of phenolic acid and organic acid. It is found in food (such as tomatoes, carrots, strawberries, blueberries and wheat), beverages (such as wine, tea, coffee and apple juice) as well as Chinese herbal medicines. In the present study, we examined the effects of caffeic acid on learning deficits in a rat model of Alzheimer's disease (AD). The rats were randomly divided into three groups: i) control group, ii) AD model group and iii) caffeic acid group. Caffeic acid significantly rescued learning deficits and increased cognitive function in the rats with AD as demonstrated by the Morris water maze task. Furthermore, caffeic acid administration resulted in a significant decrease in acetylcholinesterase activity and nitrite generation in the rats with AD compared with the AD model group. Furthermore, caffeic acid suppressed oxidative stress, inflammation, nuclear factor­κB­p65 protein expression and caspase­3 activity as well as regulating the protein expression of p53 and phosphorylated (p-)p38 MAPK expression in the rats with AD. These experimental results indicate that the beneficial effects of caffeic acid on learning deficits in a model of AD were due to the suppression of oxidative stress and inflammation through the p38 MAPK signaling pathway.

Expression and significance of angiostatin, vascular endothelial growth factor and matrix metalloproteinase-9 in brain tissue of diabetic rats with ischemia reperfusion.

OBJECTIVE: To discuss the expression and significance of angiostatin, vascular endothelial growth factor and matrix metalloproteinase-9 in the brain tissue of diabetic rats with ischemia reperfusion. METHODS: A total of 60 male Wistar rats were randomly divided into the normal group, sham group, diabetic cerebral infarction group and single cerebral infarction group according to the random number table, with 15 rats in each group. The high sucrose diet and intraperitoneal injection of streptozotocin were performed for the modeling of diabetic rats, while the thread-occlusion method was employed to build the model of cerebral ischemia reperfusion. The immunohistochemical staining was performed to detect the expression of angiostatin, vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9) in the brain tissue. RESULTS: The expression of angiostatin after the reperfusion in the brain tissue of rats in the single cerebral infarction group and diabetic cerebral infarction group was increased 6 h after the reperfusion, reached to the peak on 1 d and then decreased gradually. The expression of angiostatin in the diabetic cerebral infarction group 6 h, 1 d, 3 d and 7 d after the reperfusion was significantly higher than that in the single cerebral infarction group (P < 0.05). VEGF began to be increased 1 h after the reperfusion in the single cerebral infarction group and diabetic cerebral infarction group, reached to the peak at 6 h and then decreased gradually. The expression of VEGF in the diabetic cerebral infarction group at each time point after the reperfusion was significantly lower than that in the single cerebral infarction group (P < 0.05). MMP-9 began to be increased 1 h after the reperfusion in the single cerebral infarction group and diabetic cerebral infarction group, reached to the peak on 1 d and then decreased gradually. The expression of MMP-9 in the diabetic cerebral infarction group at each time point after the reperfusion was significantly higher than that in the single cerebral infarction group (P < 0.05). CONCLUSIONS: The high glucose environment in which the diabetic cerebral infarction is occurred is to induce the formation of MMP-9 at first and then activate and increase the expression of angiostatin. Afterwards, the expression of VEGF is inhibited, resulting in the poor angiogenesis after cerebral infarction, which thus makes the injury of brain tissue after cerebral infarction even worse than the non-diabetes mellitus.

Amelioration of ß-amyloid-induced cognitive dysfunction and hippocampal axon degeneration by curcumin is associated with suppression of CRMP-2 hyperphosphorylation.

The Alzheimer's disease (AD) brain is characterized by ß-amyloid deposition, hyperphosphorylation of microtubule-associated proteins, formation of senile plaques and neurofibrillary tangles, and degeneration of specific neuronal populations. Collapsin response mediator protein 2 (CRMP-2) hyperphosphorylation has been implicated in AD-associated neural process regression and neurofibrillary tangle formation. Curcumin is a promising AD drug with incompletely defined therapeutic mechanisms. One possibility is that curcumin prevents ß-amyloid-induced CRMP-2 hyperphosphorylation, thereby protecting against axonal regression and (or) promoting axonal regrowth. We examined spatial learning in the Morris water maze, hippocampal expression levels of CRMP-2 and phosphorylated CRMP-2 (p-CRMP-2) by Western blot, and NF-200 (an axon-specific marker) by immunohistochemistry in Sprague-Dawley rats subjected to a single intrahippocampal injection of Aß1-40 alone or Aß1-40 followed by curcumin (i.p. daily for 7 days). Compared to controls, spatial learning was significantly impaired in these Aß1-40-injected AD model rats (P<0.05). In addition, hippocampal expression levels of CRMP-2 and NF-200 were reduced while p-CRMP-2 expression was significantly enhanced (P<0.05 for all). Overexpression of p-CRMP-2 was correlated with NF-200 underexpression (r(2)=-0.67308, P<0.05), suggesting that Aß1-40 damaged hippocampal axons. Spatial learning deficits were reversed, CRMP-2 and NF-200 expression levels increased, and p-CRMP-2 expression reduced in curcumin-treated rats (all P<0.05). We propose that curcumin improves spatial learning by inhibiting CRMP-2 hyperphosphorylation, thus protecting against ß-amyloid-induced hippocampal damage or promoting regeneration.

[Effect of non-contact coculture on bone marrow stromal cells and neural stem cells].

OBJECTIVE: To observe the effect of non-contact coculture on bone marrow stromal cells (MSCs) and neural stem cells (NSCs) in neural stem cell culture medium. METHODS: MSCs and NSCs were cultured in non-contact coculture in Transwell plate, and cell morphology and immunocytochemical profile were investigated. RESULTS: In the coculture, the NSCs showed adhering growth and extended long processes, and the migrating cells formed a network of cells within 7 days. The cells differentiated into neurons, astrocytes and oligodendrocytes as shown by immunocytochemistry. Most of the MSCs grew in a non-adherent manner, giving rise to large spherical cell masses which expressed neuronal, astrocyte, and oligodendrocyte phenotypes. In the control group, the NSCs grew in suspension, some of MSCs formed small non-adherent spherical cell masses, while some cells showed adherent growth. CONCLUSION: MSCs and NSCs in the non-contact coculture can mutually promote the cell differentiation into neural cells in neural stem cell culture medium, indicating that both MSCs and NSCs can secrete some neurotrophic factors to provide a microenvironment suitable for survival and differentiation for each other.

Temporal cortex participates in spontaneous perceptual reversal.

The Necker cube is perceived as two distinct three-dimensional forms; participants experience alternation between two mutually exclusive perceptions. Perceptual dominance for one form tends to be maintained when the visual stimulus is intermittently removed. The effect is enhanced with the Necker lattice (an array of Necker cubes). Neural processes underlying perceptual reversal and stabilization are unknown. Functional MRI was used to investigate the brain regions involved. Regional activation differed between endogenous and stimulus-driven perceptual reversals, and between reversal and stabilization. Our results indicated that the right anterior portion of superior temporal sulcus is likely to be involved in perceptual stabilization (perceptual memory), whereas reversal is modulated by destabilizing influences from the right frontal lobe.

[Expressing borna disease virus phosphoprotein in PC-12 cell and its influence on the cell proliferation].

OBJECTIVE: To establish stable expressing system of Borna disease virus (BDV) phosphoprotein in PC-12 cells, and then study its influence on cell proliferation of PC-12 cells. METHOD: An expression plasmid with green fluorescence protein was cloned and identified to express BDV phosphoprotein. Cultured PC-12 cell was transfected with the recombinant plasmid by positive ion lipidsome method. Fluorescence microscopy was used to detect the expression of phosphoprotein in PC-12 cells, then G418 was added into cell culture medium to kill cells without recombinant plasmid. We performed reverse transcriptase polymerase chain reaction (RT-PCR) in the 10th generation of treated cells to examine the expression of BDV phosphoprotein. The proliferation of treated cells and control cells was examined by methyl thiazolyl tetrazolium assay (MT). RESULT: The recombinant plasmid was confirmed to be able to express BDV phosphoprotein and green fluorescence protein by both fluorescence microscopy and RT-PCR. BDV phosphoprotein expressed in PC-12 cell inhibited cell proliferation. CONCLUSION: We established a stable expressing system of BDV phosphoprotein in PC-12 cell. This cell model can be used to study the effect of BDV phosphoprotein on the centre nervous system without exposure to live virus.

[Effects of Ciwujia in inducing marrow stromal cell differentiation into neuron-like cells in vitro].

OBJECTIVE: To study the effect of the traditional Chinese herbal drug Ciwujia in inducing the differentiation of marrow stromal cells (MSCs) into neuron-like cells. METHODS: Rat MSCs isolated from the whole bone marrow were amplified by adherent culture in vitro and induced to differentiate into neuron-like cells using serum-free DMEM/F12 containing Ciwujia. The protein and mRNA expressions of nestin, beta-Tubulin III and glial fibrillary acidic protein (GFAP) in the differentiated cells were detected by indirect immunofluorescence method, Western blotting and RT-PCR. RESULTS: The third-passage MSCs showed positive expression rates for CD44 and CD54 beyond 90% with decreased CD14 expression rate to 2.37%. Induction by Ciwujia of the MSCs resulted in cell body shrinkage and protrusion of the cell processes resembling those of neurons. The differentiated cells were positive for nestin and beta-Tubulin III expression and negative for GFAP as shown by immunofluorescence assay, Western blotting and RT-PCR. CONCLUSION: Ciwujia can induce the differentiation of rat MSCs into neuron-like cells in vitro.

[Study of mechanism on differentiation of marrow stroma cells into neuron-like cells induced by Ciwujia injection in vitro].

OBJECTIVE: To study whether the nuclear factor-kappa B (NF-kappaB) play a role in differentiation of marrow stroma cells (MSCs) into neuron-like cells induced by Ciwujia injection in vitro. METHOD: Rats were randomly divided into control group and Ciwujia induced group. Cell morphology was observed with invert microscope; MSCs phenotype, expression of neural marker protein and nucleus translocation of NF-kappaB subunit RelA (p65) after induction were observed by immunofluorescence; Expression of IkappaBalpha in Ciwujia induced group and control group was detected by Western blot. RESULT: The decreased volume of some MSCs, strengthened three-dimensional structure and shrinkage of cell body with formation of spire or round was observed after induction by Ciwujia for 1 hour. After 5-hour exposure to Ciwujia, changes in the morphology of MSCs were as follows: appearance of more triangular or round shapes cells, retraction of microfilament in MSCs, formation of slender process from pseudopod with reticular structure, exfoliation and necrosis. After 7-day induction, most MSCs became triangular and round shapes, exhibiting a typical neuronal structure. Immunofluorescence method showed that nestin expression displayed intensively positive in Ciwujia group at 1 hour, and still remain positive at 7 day. However, beta-III Tubulin expression was weakly positive at 1 hour and intensive positive at 3 and 5 hour. It was still positive at 7 day. NSE expression was weakly positive at 3 hour and intensive positive at 5 hour and 7 day. GFAP expression was negative. However, in the control group, every expression of marker protein was negative or weakly positive. Expression of p65 was in cytoplasm. Western blot method showed IkappaBalpha protein relative IA was 1.000 before induction. It was 0.4556 +/- 0.1008 after induction in Ciwujia group and decreased significantly than before induction (P < 0.01). It was 0.0296 +/- 0.0099 after induction in control group and decreased significantly than before induction and in Ciwujia group (P < 0.01). CONCLUSION: The data showed that MSCs can induce differentiation into neuron-like cells by Ciwujia, and Ciwujia could inhibit the translocation of NF-kappaB from the cytoplasm to the nucleus, which may be relate to the differentiation of MSCs into neuron-like cells.

A simple and efficient method for deriving neurospheres from bone marrow stromal cells.

Bone marrow stromal cells (MSCs) can be differentiated into neuronal and glial-like cell types under appropriate experimental conditions. However, previously reported methods are complicated and involve the use of toxic reagents. Here, we present a simplified and nontoxic method for efficient conversion of rat MSCs into neurospheres that express the neuroectodermal marker nestin. These neurospheres can proliferate and differentiate into neuron, astrocyte, and oligodendrocyte phenotypes. We thus propose that MSCs are an emerging model cell for the treatment of a variety of neurological diseases.