Preliminary study of bone marrow mesenchymal stem cell-derived exo-somes in repair of spinal cord injury in rats / 中国病理生理杂志

AIM:To study the influence of bone marrow mesenchymol stem cell-drived exosomes(BMSC-exo-somes)on hindlimb activity,and the numbers of reactive astrocytes and residual neurons in spinal cord injury(SCI)rats. METHODS:BMSCs were cultured using the whole bone marrow adherent culture method and surface markers CD 90 and CD34 were verified by flow cytometry.Exosomes were isolated by ultracentrifugation and the morphology of exosomes was observed under transmission electron microscope.The protein markers CD63 and CD9 were verified by Western blot.After exosomes were applied to SCI rats,the Basso,Beattie and Bresnahan locomotor rating scale score,the Nissl staining of the lesion site,and the numbers of reactive astrocytes and residual neurons were assessed at various time points.RESULTS:Transmission electron microscopic observation revealed the presence of saucer -shaped vesicles.BMSC-exosomes were found to express high levels of CD63 and CD9.Compared with injury group,significant improvement of hindlimb activity scores from day 14 after injury in treatment group was observed(P<0.05),and less reactive astrocytes and more residual neu-rons from day 7 after injury were also observed(P<0.05).CONCLUSION:BMSC-exosomes inhibit reactive astrocytes and death of neurons,and improve hindlimb activity in the rats after SCI.

Effect of p65 gene inhibited by siRNA on differention of rat marrow mesenchymal stem cells into neurons / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate the effect of p65 gene inhibited by siRNA on neuronic differentiation in the marrow mesenchymal stem cells (MSCs).</p><p><b>METHODS</b>The MSCs were transfected with Rn-p65-siRNA. Fasudil hydrochloride induced MSCs differentiating into neurons. The non-transfected group and negative control group (transfected with negative control siRNA marked by Cy3) were used as controls. The fluorescence expressed by transfected MSCs were observed under inverted fluorescence microscope at 24 h,48 h and 72 h after transfected with negative control siRNA. The viability of MSCs was detected by MTT at 24 h, 48 h and 72 h after transfected with Rn-p65-siRNA. The expressions of p65 mRNA and protein in MSCs were detected by RT-PCR and Western blot respectively. The expressions of p65 protein, NSE, MAP-2 and glial fibrillary acidic protein (GFAP) were detected by immunocytochemical method after transfection for 6 h.</p><p><b>RESULTS</b>The fluorescence of MSCs was mostly displayed after transfection of 72 hours and the efficiency of transfection was up to 83.3% ± 3.8%. Meanwhile, the p65 mRNA and p65 protein expressed by MSCs of transfected group were significantly decreased (P < 0.05); MTT displayed that the viability of MSCs was also significantly reduced (P < 0.05). The best efficiency of induction was observed in the transfected group. There were higher expressions of NSE and MAP-2 than the other group (P < 0.05).</p><p><b>CONCLUSION</b>The p65 gene inhibited by siRNA can promote the marrow mesenchymal stem cells to differentiate into neurons.</p>

A preliminary study of plasma microRNA levels in children with methylmalonic acidemia / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To screen out differentially expressed microRNAs (miRNAs) in the plasma of children with methylmalonic acidemia (MMA), to determine the expression of miR-9-1 in plasma and to preliminarily evaluate the significance of miR-9-1 as a biomarker in MMA.</p><p><b>METHODS</b>Plasma was obtained from 17 MMA children, 10 hyperhomocysteinemia (HHcy) children without MMA (HHcy group), and 10 normal controls. Of 17 MMA children, 12 had HHcy (MMA+HHcy group), and 5 had no HHcy (MMA group). The differentially expressed miRNAs were screened out by miRNA microarray. Differentially expressed miR-9-1 was selected, and plasma miR-9-1 levels were determined by RT-PCR. Urine was collected from MMA patients who received vitamin B12 treatment, and plasma miR-9-1 levels were determined by RT-PCR after treatment.</p><p><b>RESULTS</b>The miRNA microarray analysis showed that 26 miRNAs were differentially expressed, among which 16 miRNAs (including miR-9-1) were down-regulated over 2 times, while 10 miRNAs were up-regulated over 2 times. The MMA+HHcy , MMA and HHcy groups had significantly down-regulated miR-9-1 compared with the normal control group (P<0.01). The patients who showed a good response to vitamin B12 treatment had significantly increased plasma miR-9-1 levels, without significant difference compared with the normal control group.</p><p><b>CONCLUSIONS</b>Plasma miR-9-1 is significantly down-regulated in MMA patients, but it is significantly up-regulated after vitamin B12 treatment, suggesting that miR-9-1 may act as a biomarker in monitoring the progression of MMA.</p>

miR-124-1 promotes neural differentiation of rat bone marrow mesenchymal stem cells / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the effects of miR-124-1 on neuronal differentiation of rat bone marrow mesenchymal stem cells (MSCs).</p><p><b>METHODS</b>MSCs cells were assigned into three groups: control (uninfected and untransfected), miR-124-1+ (infected with miR-124-1), and miR-124-1- (transfected with Anti-rno-miR-124* Inhibitor). MSCs were induced by β-mercaptoethanol (β-ME) to differentiate into neurons. The fluorescence expressed by infected MSCs was observed under an inverted fluorescence microscope. MTT method was used to measure cell survival rate after transfection or infection. Immunocytochemistry, RT-PCR and Western blot methods were used to detect the expression of β3 tubulin, MAP-2 and GFAP 6 days after β-ME induction.</p><p><b>RESULTS</b>The expression of miR-124-1 in the miR-124-1+ group was significantly higher 2 days after infection of lentivirus vector compared with the control group (P<0.01). In the miR-124-1- group, the cell survival rate and the miR-124-1 expression level decreased significantly 24 hrs after transfection of anti-rno-miR-124* inhibitor (P<0.01). After 6 days of β-ME induction, the protein and mRNA expression levels of β3 tubulin and MAP-2 in the miR-124-1+ group were much higher than the other two groups (P<0.01); while the expression levels of β3 tubulin and MAP-2 in the miR-124-1-group were lower than the control group (P<0.01). The expression of GFAP in the three groups was weak (<1%).</p><p><b>CONCLUSIONS</b>miR-124 might promote neuronal differentiation of rat MSCs.</p>

Effect of miRNA-9 modified marrow mesenchymal stem cell transplantation on treatment of rats with injured spinal cord / 中华神经医学杂志

Objective To assess the role of microRNA-9 in bone marrow mesenchymal stem cells differentiating into neurons and research the role of gene modification in spinal cord injury treatment.Methods Adherent culture was used to isolate and culture rat bone marrow mesenchymal stem cells (MSCs); microRNA-9-1 lentiviral vector was constructed.Acute spinal cord injury (SCI) models were established in 84 adult SD rats at T10/11 level according to the improved Allen's method; then,they were randomly divided into control group,MSCs group and miRNA group (n=28).One week after SCI,the rats of MSCs group were treated with MSCs implantation,and the rats of miRNA group were treated with MSCs-transfected microRNA-9-1 lentiviral vector; while rats of the control group only received the same amount of physical saline in the same region.The neurological functions were evaluated by using Basso-Beattie-Bresnahan (BBB) scale 1 and 3 days,and 1,2,4,6,8 and 12 weeks after SCI.The immunoreactivity of neuro filament 200 (NF-200) and glial fibrillary acidic protein (GFAP) was measured,and the percentage of positive response area was assayed and compared between groups.Results Four weeks after cell transplantation,statistical difference of BBB scale scores was noted between each two groups; the scores of miRNA group were significantly higher than those of the MSCs group and control group (P＜0.05).The immunohistochemistry staining indicated that the expression of NF-200 was significantly more intense and the expression of GFAP was significantly weaker in miRNA group than those of the other two groups (P＜0.05).Conclusion MicroRNA-9 may play an important role in bone marrow mesenchymal stem cells differentiating into neurons,and possess effects on repairing injured spinal cord and promoting functional recovery through promoting axonal regeneration and reducing the number of reactive glial cells in the spinal cord injury site.

Significance of soluble DLL1 in diagnosis of intracranial infectious diseases in children / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the significance of soluble DLL1 (Delta-like-1) levels of cerebrospinal fluid (CSF) and serum in the diagnosis of intracranial infection in children.</p><p><b>METHODS</b>Fifty children with intracranial infection, including 20 cases of tuberculous meningitis (TM), 20 cases of viral meningitis (VM) and 10 cases of purulent meningitis (PM), and 20 children without intracranial infection (control group) were enrolled. The levels of soluble DLL1 in CSF and serum were measured using ELISA.</p><p><b>RESULTS</b>The level of CSF soluble DLL1 in the TM group was significantly higher than that in the VM, PM and control groups (2.89 ± 1.72 ng/mL vs 0.14 ± 0.14 ng/mL, 0.27 ± 0.21 ng/mL, 0.13 ± 0.12 ng/mL; P<0.01). The level of serum soluble DLL1 in the TM group was also significantly higher than that in the VM, PM and control groups (12.61 ± 6.45 ng/mL vs 2.28 ± 2.27 ng/mL, 2.38 ± 1.79 ng/mL, 2.26 ± 2.10 ng/mL; P<0.01). The levels of soluble DLL1 in the CSF and serum in the VM and PM groups were not significantly different from those in the control group.</p><p><b>CONCLUSIONS</b>Soluble DLL1 as a novel indicator might have potentially important value in the diagnosis of TM.</p>

Effect of notch signaling on differentiation of rat marrow mesenchymal stem cells into neurons induced by fasudil hydrochloride / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate the effect of notch signaling on differentiation of rat bone marrow mesenchymal stem cells (MSCs) into neurons induced by fasudil hydrochloride.</p><p><b>METHODS</b>The experiments were divided into non-transfected group, transfected group (transfected with Rn-Notch1-siRNA), positive control group (transfected with Rn-MAPK-1 Control siRNA) and negative control group (transfected with negative control siRNA). Fasudil hydrochloride induced MSCs differentiating into neurons. The fluorescence expressed by transfected MSCs were observed under inverted fluorescence microscope. The expression of notch1 mRNA, Hes1 mRNA and MAPK1 mRNA in MSCs was detected by RT-PCR. The expression of Notch1 protein, NSE, neurofilament M (NF-M) and glial fibrillary acidic protein(GFAP)was detected by immunocytochemical method. The viability of MSCs was detected by MTT.</p><p><b>RESULTS</b>(1) The fluorescence of MSCs was mostly displayed after transfection for 72 h and the efficiency of transfection was up to 91.3% +/- 4.2%. Meanwhile, the notch1 mRNA and Hes1 mRNA expressed by MSCs of transfected group were significantly decreased (P < 0.05) and MTT displayed that the viability of MSCs was also significantly reduced (P < 0.05). (2) Fasudil hydrochloride could induce MSCs differentiate into neurons and the best efficiency of induction was observed in the transfected group. There was higher expression of NSE and neurofilament-M (NF-M) than the other groups (P < 0.05).</p><p><b>CONCLUSION</b>There may be notch1 signaling and Rho/Rho GTPase signaling synergy on differentiation of rat bone marrow stromal cell into neurons induced by fasudil hydrochloride and they jointly promote the differentiation of MSCs into neurons.</p>

Clinical analysis of 322 cases of non-epileptic cerebral palsy / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the clinical features of non-epileptic seizures associated with cerebral palsy (CP) in children.</p><p><b>METHODS</b>A total of 1 198 children with CP (age: 9 months to 6 years) were enrolled. The children with paroxysmal events were monitored by 24 hrs video-EEG (VEEG) to make sure the seizures were epileptic or non-epileptic. The symptoms, age, CP types and EEG features were observed in children with non-epileptic CP.</p><p><b>RESULTS</b>Five hundred and seventy-eight children (48.24%) presented paroxysmal events. The seizures were epileptic in 231 children (19.28%) and non-epileptic in 322 cases (26.88%). In the 322 cases of non-epileptic CP, the paroxysmal events were of various kinds, including non-epileptic seizure tonic, seizure shake head, shrug shoulder or head hypsokinesis, cry or scream, panic attacks, sleep myoclonic and stereotyped movement. One hundred and fifty-eight (49.1%) out of the 322 children demonstrated nonspecific EEG abnormalities. One hundred and eleven children (34.5%) were misdiagnosed as epilepsy in primary hospitals. The CP children less than one year old showed higher frequency of non-epileptic seizures than the age groups over 1 year and 3 to 6 years. The frequency of non-epileptic seizures was the highest in children with spastic CP (168 cases, 52.2%), followed by dyskinetic CP (69 cases, 21.4%) and mixed type CP (65 cases, 20.2%).</p><p><b>CONCLUSIONS</b>The paroxysmal events in children with CP partially are non-epileptic seizures and it is important to differentiate non-epileptic from epileptic seizures. The frequencies of non-epileptic seizures may be associated with a child's age and CP type.</p>

Protective effect of musk extract on rat's cerebral cortical neurons with inflammatory injury / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the protective effects of musk extract (ME) and its possible mechanism on rat's cerebral cortical neurons with inflammatory injury induced by lipopolysaccharide (LPS).</p><p><b>METHODS</b>Neurons and astrocytes from newborn rat cerebral cortex were cultured in vitro respectively, and the astrocyte conditioned medium (ACM), obtained by treating astrocytes with 10 mg/L LPS and different concentrations of ME for 24 h, was added in the culture fluid of neurons. The survival rate and apoptotic rate of neurons were measured by MTT method and AO/EB stain; and the changes of inflammatory factors in the ACM were determined by ELISA.</p><p><b>RESULTS</b>The survival rate (%) of neurons treated by ACM with ME in concentrations of 18 mg/L, 36 mg/L, 72 mg/L and 144 mg/L was 52.55 +/- 3.52, 55.77 +/- 2.36, 64.89 +/- 3.45 and 73.67 +/- 1.80, respectively, significantly higher than that in the model neurons (43.62 +/- 4. 51, P < 0.05), while the apoptotic rate (%) in them, 68.11 +/- 2.16, 44.27 +/- 3.68, 32.56 +/- 2.14 and 21.89 +/- 2.46, respectively, was significantly lower than that in model neurons (71.33 +/- 3.25, P < 0.05 or P < 0.01). Level of IL-6 was decreasing along with the raising of ME concentration in the ACM, showing a concentration-dependent state.</p><p><b>CONCLUSION</b>ME shows apparent protective effect on neurons against inflammatory injury, especially in a high concentration (144 mg/L), which may be associated with the reduction of IL-6 secreted by astrocytes.</p>

Effect of non-contact coculture on bone marrow stromal cells and neural stem cells / 南方医科大学学报

<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>MSCs and NSCs were cultured in non-contact coculture in Transwell plate, and cell morphology and immunocytochemical profile were investigated.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>

Expression of microRNA in neonatal rats with hypoxic-ischemic brain damage / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the changes of microRNA expression in cortex tissues in neonatal rats with hypoxic-ischemic brain damage (HIBD)and the possible roles of microRNA in the pathogenesis of HIBD. METHODS Rat HIBD model was prepared. The cortex tissues were obtained 14 days after the HIBD event. The microRNA expression profiles were measured using microRNA microarray. Expression of 9 microRNAs (miR-126,-26a,-674-5p,-21,-25,-290, miR-124,-125b-5p and microRNA-9a) was determined by quantitative real-time PCR.</p><p><b>RESULTS</b>he results of microRNA expression profiles indicated that 27 pieces of microRNA were up-regulated more than 2 folds and 60 pieces were down-regulated more than 2 folds compared with the normal control group. The results of the 9 microRNAs detected by quantitative real-time PCR were consistent with those detected by microRNA microarray.</p><p><b>CONCLUSIONS</b>HIBD rats have significant changes in microRNA expression, suggesting that microRNA expression may play important roles in the pathogenesis of HIBD.</p>

Effects of DSCAM on differentiation of rat marrow mesenchymal stem cells into neurons in vitro / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the effects of Down syndrome cellular adhesion molecule (DSCAM) on differentiation of rat marrow mesenchymal stem cells (MSCs) into neurons in vitro.</p><p><b>METHODS</b>MSCs from Sprague-Dawley rats were induced into neurons by baicalin. The expression of DSCAM before and after induction was evaluated by immunocytochemical staining and Western blot assay. After knockdown of DSCAM by siRNA transfection, the differentiation rate of neurons derived from MSCs was measured.</p><p><b>RESULTS</b>Before induction, the expression of DSCAM was not detectable in MSCs. After bFGF preinduction for 24 hrs, DSCAM was slightly expressed in MSCs (1.71+/- 0.67%). The DSCAM expression increased 6 hrs after baicalin induction (15.79+/- 4.24%), reached a peak at 3 days (53.16+/- 5.94%) and then decreased gradually. The DSCAM expression 6 days after baicalin induction (28.99+/- 6.72%) was significantly lower than that at 3 days (P<0.01). However, after DSCAM-siRNA transfection, the DSCAM expression in MSCs was significantly reduced. MSCs did not express neuron-specific beta-III-tubulin before induction. After baicalin induction for 6 hrs, 3 days and 6 days, the expression of beta-III-tubulin was 1.40+/- 0.79%, 41.59+/- 3.17% and 59.11+/- 4.76% respectively. But the beta-III-tubulin expression significantly decreased 3 and 6 days after DSCAM-siRNA transfection (28.57+/- 2.91% and 43.90+/- 12.31% respectively).</p><p><b>CONCLUSIONS</b>DSCAM may play an important role in MSCs differentiation into neural cells.</p>

Clinical analysis of families with generalized epilepsy with febrile seizures plus / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the clinical phenotypes and hereditary patterns of the generalized epilepsy with febrile seizures plus (GEFS+).</p><p><b>METHODS</b>Detailed family trees were constructed by inquire and physical examinations for the probands of the 15 pedigrees of GEFS+. Some patients received electroencephalography, cranial CT or MRI examination. The seizures and epilepsy syndromes were classified according to the 2001 Seizure International Classification. The clinical data of GEFS+ were reviewed.</p><p><b>RESULTS</b>The 15 families consisted of 196 individuals. Seventy-five individuals were confirmed with epilepsy. The phenotypes of 64 out of the 75 patients with epilepsy conformed to GEFS+. The 64 patients included 38 males and 26 females (1 deceased) and there was no gender difference in the morbility of GEFS+. The age at onset was all in childhood. GEFS+ had a diversity of phenotypes. Febrile seizures (FS) were confirmed in 44 patients, FS and myoclonic seizure in 1, febrile seizures plus (FS+) in 13, FS+ and absence seizure in 2, FS+ and myoclonic seizure in 1, and FS+ and focal seizure in 3.</p><p><b>CONCLUSIONS</b>The heterogeneity of phenotypes and genetics may be the hallmarks of GEFS+. FS and FS+ are common phenotypes while FS+ and absence seizure, FS+ and myoclonic seizure, and FS+ and focal seizure are rare. If one of the parents is affected in a GEFS+ family, the susceptibility of their children to GEFS+ is the same no matter what gender of their children is. It is speculated that the hereditary pattern of GEFS+ conforms to autosomal dominant inheritance.</p>

Effect of hyperbaric oxygenation on neural stem cells and myelin in neonatal rats with hypoxic-ischemic brain damage / 中国当代儿科杂志

<p><b>OBJECTIVE</b>This study investigated the effect of hyperbaric oxygenation (HBO) on neural stem cells (NSCs) and myelin in neonatal rats following hypoxic-ischemic brain damage (HIBD) and aimed to explore the possible mechanism of the protective effect of HBO on HIBD.</p><p><b>METHODS</b>Seven-day-old Sprague-Dawley rat pups were randomly assigned into 4 groups: Normal control, HIBD, hyperbaric air (HBA), and HBO groups (n=30 each). The HIBD model was produced by permanent occlusion of the left common carotid artery and 2 hrs hypoxemia exposure (8% O2 at 37 degrees C). HBA and HBO treatment was administered (2 ATA, once daily for 7 days) in the HBA and HBO groups respectively 1 hr after HIBD. BrdU immunohistochemistry was used to detect the NSCs in the sub-ventricle zone (SVZ) of the lateral ventricle and the dentate gyrus (DG) of the hippocampus. The myelin damage was assessed by myelin basic protein (MBP) immunostaining.</p><p><b>RESULTS</b>The BrdU-positive cells in the SVZ and the DG of the ischemic hemisphere in the HIBD group were dramatically decreased compared with those of the Normal control group at 3 weeks post-HIBD (P < 0.01). The HBO treatment resulted in an increase of BrdU-positive cells in the DG from 153.7 +/- 37.0 to 193.7 +/- 38.8 (P < 0.05). The nestin expression in the HIBD and HBA groups was reduced compared with that in the Normal control group. There was no difference in the nestin expression between the HBO and the Normal control groups. Hypoxia-ischemia (HI) led to marked myelin damage at 1 week post-HIBD. HBO or HBA treatment alleviated the damage.</p><p><b>CONCLUSIONS</b>The HBO treatment can result in the proliferation of BrdU-positive cells and alleviate the myelin damage following HIBD in neonatal rats, thereby offering neuroprotectivity against HI insults.</p>

Study on molecular mechanism of inhibitory effect of baicalin on proliferation of insulinoma cell line in rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the effect of baicalin on insulinoma cell line and the molecular mechanism involved.</p><p><b>METHODS</b>Light microscope, MTT assay, flow cytometry, gene analysis and Western Blot were applied to investigate the effects of baicalin on the cell proliferation, the cell cycle and the involved molecular mechanism.</p><p><b>RESULTS</b>After treatment with baicalin, the number of cells in mitotic stage and the survival rate of cells obviously decreased, and cell proliferation was inhibited in a drug concentration- and acting time-dependent manner, with the appearance of apoptotic insulinoma cells. During the apoptotic process, the activity of caspase-3 was elevated by baicalin in a time-dependent manner; with the increase of the concentration of baicalin, the number of cells in S-phase obviously decreased from 38.2% to 9.4%, while the percentage of cells in G0/G1 phase increased from 56.4% to 85.9%, indicating cells were arrested in G1-phase. Meanwhile, the activity of cyclin gene promoter obviously declined, and the expression of cyclin reduced remarkably.</p><p><b>CONCLUSION</b>Baicalin could induce apoptosis of insulinoma cells, which might be correlated with the activity of caspase-3, and inhibiting proliferation of insulinoma cells in a concentration- and time-dependent manner, in which the action of baicalin in down-regulating the gene transcription and expression of cyclin may play an important role.</p>

Effect of nuclear factor-kappaB on differentiation of bone marrow stromal cells into neurons induced by baicalin in rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the effect of nuclear factor-kappaB (NK-kappaB) on differentiation of bone marrow stromal cells (MSC) into neurons induced by baicalin.</p><p><b>METHODS</b>Using baicalin as main inducer, differentiation of rats' MSC was induced into NC in serum-free medium, at the same time, untreated cells were cultured in serum-free medium as the control group. The expression of NC special marker protein and the presence of NK-kappaB subunits RelA (P65) translocated into nucleus were determined by immunofluorescent cytochemical stain, the change of expression of NK-kappaB inhibitory protein (IkappaBalpha) were determined by Western blot, and the apoptotic index was assessed by terminal deoxynucleotidyl transferase-mediated d-UTP-biotin nick end labeling (TUNEL) assay.</p><p><b>RESULTS</b>MSC displayed typical shape of NC and expressed several NC marker protein after induced by baicalin, while those changes were not revealed in the control group. but showed translocation of P65 from cytoplasm to nucleus, and the markedly reduction of IkappaBalpha expression, the apoptotic index being 28.2+/-6.1 %. After induced by baicalin, the P65 still revealed mainly in cytoplasm with the level of IkappaBalpha expression decreased lesser and the number of apoptosis cell (12.2+/-2.8%) lower than those in the control group, P <0.01.</p><p><b>CONCLUSION</b>Baicalin could inhibit NK-kappaB activation, and it has certain effect in inducing the differentiation of MSC into NC.</p>

Effect of baicalin on the proliferation of insulinoma cell line / 中南大学学报(医学版)

OBJECTIVE@#To investigate the effect of baicalin on the proliferation of insulinoma cell line and the molecular mechanism involved.@*METHODS@#Such methods as light microscope, MTT assay, flow cytometry and Western blotting were applied to investigate the effects of baicalin (0, 100, 200, and 400 microg/ml baicalin treated for 24 h or 200 microg/ml baicalin treated at different time points) on the cell proliferation, cell survival rate, the cell cycle and related molecular mechanisms.@*RESULTS@#The number of proliferating cells obviously decreased with the increase of baicalin under the light microscope, and the survival rate of cells decreased as determined by MTT assay. After being treated with baicalin, the number of insulinoma cells in S-phase obviously decreased from 38.2% (0 microg/ml) to 9.4% (400 microg/ml), and the number of cells in phase G1 increased from 56.4% (0 microg/ml) to 85.9% (400 microg/ml). In the meantime, the expression of cyclin D1 was obviously declined by Western blotting.@*CONCLUSION@#Baica-lin can inhibit the proliferation of insulinoma cells, and the down-regulation of the expression of cyclin D1 might also be involved in these events.

Expression of exogenous gene in bone marrow stromal cells by liposome / 中南大学学报(医学版)

OBJECTIVE@#To transform eukaryotic expression vector pEGFP-PDX-1 into marrow stromal cells by liposome and to optimize the conditions of transformation.@*METHODS@#The recombinant vector was identified by enzyme digestion analysis and sequencing. The recombinant plasmid was transformed into bone marrow stromal cells and it changed the quantity of DNA or liposome. The expression of PDX-1 gene in the transformed cells was detected by immunocytochemical staining.@*RESULTS@#Enzyme digestion analysis and sequencing showed that the interesting gene was integreted into the recombinant vector. We obtained satisfactory efficiency of transfection when the ratio of DNA and liposome was 1 : 1 or 1 : 2. The PDX-1 in the transformed cells was expressed by immunocytochemical staining.@*CONCLUSION@#The eukaryotic expression vector pEGFP-PDX-1 was constructed for the first time in China. We have enhanced the efficiency of transfection by optimizing the transformation conditions. It is possible to use the bone marrow stromal cells as seed cells in tissue-engineering.

Antagonistic effect of baicalin on oxidative stress injury in neurons and astrocytes of rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the experimental conditions for H2O2 to injure astrocytes and the effect of baicalin in protecting neurons and astrocytes from oxidative stress injury.</p><p><b>METHODS</b>Neurons and astrocytes from forebrain of rats were cultured in vitro and treated with H2O2, baicalin and combination of the two, respectively. The cell viability or survival rate was determined using MTT.</p><p><b>RESULTS</b>Effects of H2O2 in different concentrations on survival rate of astrocytes showed significant difference (F = 28.569, P < 0.01) in a dose-dependent manner. Degrees of H2O2 injury, with the same concentration of H2O2, on cells with different seeding density were also significantly different (F = 5.439, P < 0.01), and dose-dependently. Baicalin didn't influence the survival rate of neurons and astrocytes when the concentration was within 2.5-40 mumol/L (for neurons, F = 0.49, P > 0.05; for astrocytes, F = 1.001, P > 0.05), but baicalin showed significant antagonism to the injury of oxidative stress (for neurons, F = 24.384, P < 0.01; for astrocytes, F = 5.000, P < 0.01). The higher the concentration of bainalin, the higher the cell survival rate.</p><p><b>CONCLUSION</b>A model of astrocytes oxidative injury induced by H2O2 is established. Baicalin shows no toxicity on neurons and astrocytes when the concentration is within 2.5-40 mumol/L, but could antagonize the H2O2 caused oxidative injury on cells in a dose-dependent manner.</p>