Protective Effects of Anmeidan on Cell Structure Against Neuronal Damage in Hippocampal CA1 Region of Sleep-deprived Rats / 中国实验方剂学杂志

ObjectiveTo investigate the effects of Anmeidan （AMD） on neuronal structure and neuronal marker protein expression in the hippocampal CA1 region of sleep-deprived （SD） rats. MethodRats were randomly divided into control group， model group， an AMD group （9.09 g·kg-1·d-1）， and melatonin group （0.27 g·kg-1·d-1）. Rats in the control group and the model group received equal volumes of physiologicol saline. The SD model was induced by the self-made sleep deprivation box for four weeks. Ethovision XT system detected and analyzed the spontaneous behaviors of rats. The histomorphology of neurons in the hippocampal CA1 region was observed by hematoxylin-eosin （HE） staining and Nissl staining， and the changes in Nissl bodies were observed by Nissl staining. The ultrastructure of hippocampal cells was observed by transmission electron microscopy （TEM）. Immunohistochemistry was used to detect the expression of glial fibrillary acidic protein （GFAP）， microtubule-associated protein 2 （MAP2）， nestin， and neuronal nuclei （NeuN） in the CA1 region. ResultCompared with the control group， the model group showed longer distance， increased average activity speed， cumulative duration， average body fill， and higher activity frequency （P<0.01）. Besides， the neurons in the CA1 region were reduced in number with disorganized arrangement， wrinkled nuclei， deeply stained cytoplasm， reduced Nissl bodies， swollen and deformed mitochondria， shortened cristae， and swollen Golgi vesicles. Furthermore， the mean integral absorbance （IA） value of GFAP increased and those of MAP2， nestin， and NeuN decreased （P<0.01）. Compared with the model group， the AMD group showed shortened distance traveled， lower average activity speed， shorter cumulative duration， decreased average body fill， and reduced activity frequency （P<0.05， P<0.01）. Moreover， the neurons in the CA1 region were relieved from damage with increased cell number， clear nuclei and cytoplasm， increased Nissl bodies， and relieved mitochondrial damage. The IA value of GFAP decreased and those of MAP2， nestin， and NeuN increased （P<0.05， P<0.01）. ConclusionAMD can improve structural damage of neurons in the hippocampal CA1 region of sleep-deprived rats， which may be achieved by decreasing GFAP expression and increasing MAP2， nestin， and NeuN expression.

Nestin is significantly associated with the overall survival of nonsmall cell lung cancer: A meta-analysis

Aims: Some studies investigated the association between nestin and the overall survival (OS) of nonsmall cell lung cancer (NSCLC). However, the results were conflicted and inconclusive. Therefore, we performed this meta-analysis to determine the association between nestin and OS of NSCLC. Materials and Methods: PubMed and EMBASE were searched to find relevant studies. The strength of the association was calculated with the hazard ratios (HRs) and respective 95% confidence intervals (CIs). Results: High expression of nestin was significantly associated with OS of NSCLC (HR = 2.09; 95% CI = 1.59–2.77). In the stratified analysis by race, we found that the expression of nestin was significantly associated with OS of NSCLC in Asians (HR = 3.02; 95% CI = 1.80–5.07) and Caucasians (HR = 1.81; 95% CI = 1.21–2.71). In addition, when we limited the meta-analysis to studies that controlled for clinical parameters, a significant association between nestin and OS of NSCLC remained (HR = 2.19; 95% CI = 1.54–3.11). A sensitivity analysis showed no substantial modification of the estimates after exclusion of individual studies. Conclusions: In conclusion, this meta-analysis suggested that high expression of nestin was significantly associated with OS of NSCLC.

Effects of cytokine-induced neutrophil chemoattractant 3 on the survival and proliferation of neural stem cells / 中国组织工程研究

BACKGROUND: Our previous study was the first to find that bone marrow stromal cells could secrete neutrophil chemokine-3. Moreover, the expression of cytokine-induced neutrophil chemoattractant 3 (CINC-3) could be up-regulated by 1. 5 times during the differentiation of neural stem cells into neurons regulated by bone marrow stromal cells. These results suggest that CINC-3 may promote the proliferation and differentiation of neural stem cells. OBJECTIVE: To observe the effect of CINC-3 on survival and proliferation of hippocampal neural stem cells in neonatal rats. METHODS: Hippocampal neural stem cells from neonatal Sprague-Dawley rats were isolated and cultured in vitro, and passage 3 neural stem cells were divided into control and CINC-3 groups. The survival rate of the cells was measured by MTS method, cell survival and proliferation were observed using cell growth curve and live/dead cell staining, and the expression of Nestin in neural stem cells was detected by real-time PCR and immunofluorescence staining for observation of neural stem cell proliferation. RESULTS AND CONCLUSION: (1) When the concentration of CINC-3 increased from 1 to 20 μg/L, the survival rate of neural stem cells increased gradually. When the concentration of CINC-3 was 10 μg/L, the survival rate of neural stem cells was the highest and the cell viability was the best (P < 0. 05). When the concentration of CINC-3 further increased to 20 μg/L, there was no significant difference in the survival rate of neural stem cells. (2) The positive rate of Nestin in the CINC-3 group was significantly higher than that in the control group (P < 0. 05). (3) The expression of Nestin mRNA in the CINC-3 group was significantly higher than that in the control group (P < 0. 05). These findings indicate CINC-3 can promote the survival and proliferation of hippocampal neural stem cells.

Expression of Nestin and its relationship with tamoxifen curative effect and prognosis of patients with breast cancer / 国际肿瘤学杂志

Objective To investigate the expression of Nestin and its relationship with tamoxifen cura-tive effect and prognosis of patients with breast cancer. Methods A total of 82 patients with breast cancer who received tamoxifen therapy after radical mastectomy in Department of Breast Surgery of Enshi Tujia Miao Auton-omous Prefecture Central Hospital of Hubei Province from March 2009 to March 2013 were collected. And the paired cancer tissues and adjacent normal tissues preserved in liquid nitrogen were also collected. Fluorescent quantitative real-time PCR( RT-PCR)and immunohistochemistry were used to detect the content of Nestin mRNA and the expression of Nestin in breast cancer tissues. The expression of Nestin in breast cancer tissues and its relationship with clinicopathologic features were analyzed. Breast cancer cell line MCF-7 was selected, small interfering RNA(siRNA)was used to silence Nestin in MCF-7 cells,and the influence on tamoxifen sen-sitivity was observed. The relationship of Nestin and epithelial-mesenchymal transition( EMT)was detected using Western blotting. Results The results of RT-PCR indicated that the mRNA level of Nestin in breast cancer tissues was 3. 87 times as high as that in paracancerous tissues(6. 34 ± 1. 56 vs. 1. 64 ± 0. 52,t =26. 140,P < 0. 001). Immunohistochemical staining suggested that the positive rate of Nestin in breast cancer tissues was 75. 61％(62 / 82),which was significantly higher than that in paracancerous tissues[24. 39％(20 / 82)],with a significant difference(χ2 = 43. 024,P < 0. 001). The expression of Nestin was related to lymphatic vessel infiltration(χ2 = 7. 499,P = 0. 006)and lymph node metastasis(χ2 = 6. 770,P = 0. 034), and it was not related to the age of patients(χ2 = 3. 242,P = 0. 072),tumor size(χ2 = 2. 358,P = 0. 308), histological grade(χ2 = 0. 294,P = 0. 863),ductal infiltrating status(χ2 = 0. 180,P = 0. 671)and triple neg-ative breast cancer(χ2 = 0. 142,P = 0. 706). The analysis of Cox risk ratio model showed that Nestin expres-sion(HR = 1. 982,P < 0. 001;HR = 1. 537,P < 0. 001),lymphatic vessel invasion( HR = 2. 502,P <0. 001;HR = 1. 715,P < 0. 001)and lymph node metastasis(HR = 1. 818,P < 0. 001;HR = 1. 446,P <0. 001)were independent prognostic risk factors for progress-free survival and overall survival in breast cancer patients. After Nestin knockout in breast cancer cell line MCF-7,the IC50 value of MCF-7 to tamoxifen de-creased from(48. 05 ± 2. 22)μmol/ L to(35. 59 ± 2. 92)μmol/ L,with a significant difference(t = 6. 489, P = 0. 003). Silencing Nestin significantly up-regulated E-cadherin(7. 21 ± 1. 15 vs. 1. 02 ± 0. 01,t = 6. 654, P = 0. 024)and down-regulated the mesenchyme index Vimentin(0. 17 ± 0. 08 vs. 1. 01 ± 0. 02,t = 25. 015, P < 0. 001)in MCF-7 cells. Conclusion Nestin is over-expressed in breast cancer,which is associated with reduced efficacy of tamoxifen. Nestin may be a new potential prognostic biomarker for breast cancer.

Effect of ferulate-heparin-poloxamer thermosensitive hydrogels on nerve regeneration in rats with spinal cord injury / 中国药理学通报

Aim To investigate the effect of ferulate-heparin-poloxamer (SF-HP) thermosensitive hydrogels on nerve regeneration in ratswith spinal cord injury, and provide experimental basis for clinical application. Methods SD rats were randomly divided into five groups: sham group, SCI group, SCI + HP group, SCI + SF group and SCI + SF-HP group. Rats were performed moderate contusion injuries using a vascular clip for 2 min to establish SCI animal model, then rats were given BBB score and inclined plate scoring function test after SCI. BDA tracing was employed to observe the recovery of nerve conduction function. Moreover, immunohistochemical staining and Western blot-were used to measure GAP43, Nestin and GFAP levels. Results BBB score and inclined plane test score significantly increased in SF-HP treated group as compared with the model group (P < 0. 01). Staining data showed that the structure of spinal cord was void, and this phenomenon was reversed after SF-HP administration. Data showed that the level of GAP43 and Nestin increased after SF treatment (P < 0. 05), the expression of GFAP decreased (P <0. 05), and the treatment effect of SF-HP hydrogels were better than those of SF alone (P < 0. 0 5) . BDA tracing data showed that the number of positive neurons markedly increased and the repair of nerve conduction function was significantly improved in SF-HP hydrogel group compared with SF group. Conclusion HP hydrogels enhance the effect of SF on the nerve regeneration in damaged spinal cord in rats.

Expression of nestin, transcription factor SOX3 and homeobox protein DLX-4 in the facial tissue of children with cleft lip and palate / Expresión de nestina, factor de transcripción SOX3 y proteína homeobox DLX-4 en el tejido facial de niños con fisura labiopalatina

The failure of fusion of nasal and maxillary processes results in cleft lip and palate (CLP), which is one of the most common birth defects. The morphopathogenesis of this pathology is multifactorial and still largely unclear. The aim of this study was to evaluate the presence of nestin, transcriptor factor SOX3 (Sox3) and homeobox protein DLX-4 (Dlx-4) in complete unilateral (CU) and complete bilateral (CB) CLP affected facial tissue. Oral mucosa tissue samples were obtained from 17 CUCLP and 13 CBCLP patients during surgical cleft correction and 6 unaffected control subjects. Obtained tissue sections were stained with hematoxylin and eosin and by immunohistochemistry for nestin, Sox3 and Dlx-4. The intensity of staining was graded semiquantitatively. Nestin-positive structures were detected in all CUCLP and CBCLP patients' tissue samples, varying from moderate number of nestin-positive structures to numerous. Sox3 immunoreactivity was more prominent in epithelial cells in both patient groups with frequently patchy distribution. Mainly moderate number of Dlx-4-positive cells was observed in most of tissue samples. Statistically significant moderate positive correlation was found between nestin and Sox3 factors in CUCLP patient group (Spearman's rank correlation coefficient = .517, P = .034). Increase of nestinpositive structures suggests its role in the regulation of the remodeling of tissue in both CUCLP and CBCLP affected tissue. Dominance of Sox3 positivity in cleft affected epithelium indicates its possible role in (compensatory) formation of defective oral epithelium of CUCLP and CBCLP patients. The reduced expression of Dlx-4 implicates its limited regulatory role on the craniofacial development in CUCLP and CBCLP affected tissue.

La falla en la fusión de los procesos nasal y maxilar son causante de la fisura labiopalatina (FLP), que es uno de los defectos congénitos más comunes. La morfopatogenia de esta patología es multifactorial y aún poco clara. El objetivo de este estudio fue evaluar la presencia de nestina, el factor transcriptor SOX3 (Sox3) y la proteína homeobox DLX-4 (Dlx-4) en todo el tejido facial afectado por FLP bilateral unilateral (FU) y bilateral completa (FB). Se obtuvieron muestras de tejido de mucosa oral de 17 pacientes FUFLP y 13 FBFLP durante la corrección quirúrgica de la fisura y de 6 sujetos de control no afectados. Las secciones de tejido obtenidas se tiñeron con hematoxilina y eosina y mediante inmunohistoquímica para nestina, Sox3 y Dlx-4. La intensidad de la tinción fue graduada semicuantitativamente. Se detectaron estructuras positivas para nestina en todas las muestras de tejido de pacientes FUFLP y FBFLP, variando desde un número moderado a numerosas estructuras positivas para nestina. La inmunorreactividad de Sox3 fue más prominente en las células epiteliales en ambos grupos de pacientes con distribución frecuentemente irregular. Se observó un número principalmente moderado de células Dlx-4-positivas en la mayoría de las muestras de tejido. Se encontró una correlación positiva moderada estadísticamente significativa entre los factores de nestina y Sox3 en el grupo de pacientes FUFLP (coeficiente de correlación de rangos de Spearman = 0,517, P = 0,034). El aumento de estructuras positivas para nestina sugiere su papel en la regulación de la remodelación de tejido, tanto en tejido afectado por FUFLP como FBFLP. La dominancia de la positividad de Sox3 en el epitelio afectado de la fisura, indica su posible papel en la formación (compensatoria) del epitelio oral defectuoso de pacientes FUFLP y FBFLP. La expresión reducida de Dlx-4 implica su función reguladora limitada en el desarrollo craneofacial en tejido afectado por FUFLP y FBFLP.

Effect of Moxibustion on Delayed Memory and Expression of Hippocampal Nestin and Doublecortin Proteins in Dementia Rats / 针刺研究

OBJECTIVE: To observe the effect of "Huayu Tongluo"(Blood-stasis Dispersing and Meridian-collateral Dredging) moxibustion on the delayed memory and expression of Nestin and Doublecortin (DCX) proteins in the hippocampus in vascular dementia (VD) rats in the view of neurogenesis produced by intracerebral transplantation of neural stem cells (NSCs) and endothelial progenitor cells (EPCs). METHODS: Healthy male Wistar rats were randomized into control group, VD model group,NSCs＋EPCs group and NSCs＋EPCs moxibustion group. The VD model was established by using a modified 2-vessels occlusion method, and neurogenesis was produced by transplantation of NSCs＋EPCs (2×106cell/10 µL) into the lateral ventricle for rats of the NSCs＋EPCs groups 3 days after successful VD-modeling. Moxibustion was applied to "Dazhui" (GV 14), "Baihui" (GV 20) and "Shenting" (GV 24) once daily for 21 days with an interval of one day between every two 7 days. The Morris Water Maze was used to test the rat's delayed memory ability before and 24 h after the treatment. The expression of Nestin and DCX proteins in the hippocampus tissues was detected using double-labeled immunofluorescence technique. RESULTS: Following modeling, Morris Water Maze tests showed that the average escape latency of location navigation task was significantly prolonged in VD rats(P<0.008)and the times of target platform crossing (spatial probing task) within 120 s were remarkably reduced in VD rats (P<0.008). Compared with pre-treatment in the same one group, the escape latency of NSCs＋EPCs and NSCs＋EPCs moxibustion groups were considerably reduced (P<0.05), and the average times of target platform crossing of the NSCs＋EPCs moxibustion group were markedly increased(P<0.05). The effect of NSCs＋EPCs moxibustion was evidently superior to that of simple NSCs＋EPCs in shortening the escape latency (P<0.008). The expression levels of Nestin protein were significantly higher in the NSCs＋EPCs moxibustion group after 1 and 3 period treatment than those in the NSCs＋EPCs group (P<0.05)．. CONCLUSION: Moxibustion intervention is able to improve the delayed memory in VD rats, which may be related to its effect in up-regulating the expression of hippocampal Nestin and DCX proteins within 15 days via accelerating neurogenesis.

Comparative study on extraction and culture method of neural stem cells / 中国药理学通报

Aim To explore the best method of neural stem cellextraction and culture, and provide a technical reference for thebasic research of neural stem cells. Methods Different extractionand culture methods of neural stem cells were compared.The rate of ball of neural stem cells and the stability of neuralstem cells in undifferentiated state were observed by extraction offetal and neonatal rats cortex, using different types of medium,different inoculation density, different culture methods, differentmethods of changing liquid and different passage methods. Neuralstem cells' activities were detected by Varioskan LUX MultimodeMicroplate Reader. Results ① The brain cortex of fetalrats of 14 d had higher proportion of neural stem cells, less othercells and more neurospheres than newborn rats of 24 h. ② Neuralstem cells could be stabilized in undifferentiated state by usingserum-free medium, while most of the neural stem cellswere differentiated into neurons and glial cells by using serummedium. ③ Neural stem cells, seeding at 1. 0 ×109 ·L-1 , hada large number of neurosperes and were in good condition. ④Suspension culture was beneficial to form a stable neurosphereand keep the neural stem cells in an undifferentiated state thanadherent culture. ⑤ The state of neurosphere by changing halfof the medium and adding medium without discarding was betterthan that by replacing all medium. ⑥ The neurospheres couldbe separated into single cells by mechanical blow in primary generationand the second generation of neurospheres cultured in serum-free medium. While the percentage of viable cells in neuralstem cells was the highest digested with stem cell lysates afterthree generations and the neurospheres re-formed were better. ⑦Neural stem cells' activity of 14 d fetal rat in Accutase digestiongroup was significantly higher than that of the other threegroups, and the difference was significant (P <0. 05). ConclusionsNeural stem cells proliferate and divide well, with highrate of ball formation and good neurosphere condition, which canmaintain a stable undifferentiated state by extracting the cerebralcortex of 14 d fetal rats, using serum-free medium, inoculatinginto a 25 cm2 flask at a density of 1. 0 × 109 ·L-1 , and takingthe suspension culture (adding the medium 1 ~ 1. 5 mL every 2~3 d and passage every 6 ~8 d ).

Neuroprotective effect of erythropoietin on rat model of hypoxic-ischemic encephalopathy / 中国比较医学杂志

Objective To study the protective effect of recombinant human erythropoietin (EPO) on brain and to explore the changes in the diversity of intestinal microbial flora in neonatal rats with hypoxic-ischemic encephalopathy (HIE) by establishing a neonatal rat model of HIE, and to provide an experimental basis for clinical application of EPO in the treatment of neonatal HIE. Methods The HIE model was established in 7-day-old neonatal SD rats. The rats were randomly divided into the HIE model group, EPO-treated group and control group. The changes of nestin expression were detected by immunohistochemistry. Feces of the rats were collected to detect the changes in intestinal microbial flora by 16s rRNA sequencing. Results The expressions of nestin at the same time point in each group were significantly different (P ＜0. 05). The nestin level in the control group was the lowest, that in the EPO-treated group was the highest, and the HIE model group in between. The Shannon-Wiener index of the HIE model group showed a tendency to decrease compared with the control group. Conclusions Exogenous EPO can promote the growth of neural cells in neonatal rats with HIE, indicating a certain protective effect. Meanwhile, the diversity of intestinal microbial flora of the HIE neonatal rats is also changed.

Retinal ganglion cell-conditioned medium alters the differentiation of rat retinal progenitor cells / 眼科新进展

Objective To investigate the effects of retinal ganglion cell-conditioned medium on the differentiation of retinal stem cells.Methods Rettial stem cells and retinal ganglion cells were isolated from rats,and immunofluorescence staining was applied to identify rat retinal stem cells and retinal ganglion cells with Nestin and Thy-1 antibody,respectively.Retinal stem cells were cultured in presence or absence of retinal ganglion cell-conditioned medium for 72 h,followed by detection of Nestin,PAX6,Thy-1 and Bin-3 gene expression in retinal stem cells by qPCR.Results isolated retinal stem cells were Nestin positive,and retinal ganglion cells were Thy-1 positive,indicating the success of isolation.Compared to retinal stem cells cultured without ganglion cellconditioned medium,ones cultured with ganglion-conditioned medium had significantly downregulated expression of Nestin and PAX6 (both P ＜ 0.000 1),and markedly upregulated expression of Thy-1 and Brn-3 (both P ＜ 0.05).Conclusion Retinal ganglion cell-conditioned medium can induce the differentiation of retinal progenitor cells into retinal ganglion-like cells.

Progress of hematopoietic microenvironment / 白血病·淋巴瘤

The hematopoietic microenvironment provides an important place for hematopoietic stem cell (HSC) to self-renew, directional differentiation and maintain relative homeostasis, and it can regulate hematopoietic activity through various cellular components and factors. HSC is a primitive pluripotent stem cell in the blood system featured by the potential ability to self-renew for a long time and to differentiate into various mature blood cells, which exists in the bone marrow (BM). HSC plays an important role in regulating and maintaining the physiological balance of various cellular components of the blood system in the body, to ensure the continuous regeneration of the blood system. The hematopoietic microenvironment affects the development of HSC all the time, but the related cellular molecules and signals in the microenvironment still need to be studied in depth. This paper reviews the main components of the hematopoietic microenvironment, signaling pathways and the effects of abnormal changes on the diseases.

Effect of oxygen glucose deprivation-reperfusion in astrocytes overexpressing endothelin-1 on the proliferation of neural stem/progenitor cells / 器官移植

Objective To investigate the effect of oxygen glucose deprivation-reperfusion (OGD-R) in astrocytes overexpressing endothelin (ET)-1 on the proliferation of neural stem/progenitor cells (NSPCs). Methods OGD-R models of negative control astrocytes (C6-Mock) and astrocytes over-expressing ET-1 (C6-ET-1) were constructed. Transwell co-culture system of astrocytes and NSPCs was established. Morphologic observation and identification of the astrocytes and primary NSPCs were performed. The cells were divided into four groups: C6-Mock+NSPCs, OGD-R+C6-Mock+NSPCs, C6-ET-1+NSPCs and OGD-R+C6-ET-1+NSPCs groups and co-cultured for 0, 24, 48 and 72 h respectively. The diameter of neurosphere was measured in each group. Results In the C6-Mock and C6-ET-1 cells, type Ⅰ astrocytes in fibrous morphology were observed. Glial fibrillary acidic protein (GFAP) was expressed in the cytoplasm of these two types of cells. Primary NSPCs were positive for nestin staining. After co-culture for 48 and 72 h, the neurosphere diameter in the OGD-R+C6-Mock+NSPCs group was significantly greater than that in the C6-Mock+NSPCs group. The neurosphere diameter in the OGD-R+C6-ET-1+NSPCs group was considerably greater than that in the C6-ET-1+NSPCs group. The neurosphere diameter in the OGD-R+C6-ET-1+NSPCs group was significantly greater compared with that in the OGD-R+C6-Mock+NSPCs group (all P<0.05). Conclusions OGD-R astrocytes can promote the proliferation of NSPCs. ET-1 over-expression further accelerates the proliferation of NSPCs.

Protective mechanism of ginsenoside Rg1 on hippocampus of aging mice induced by D-galactose / 中草药

Objective To investigate the protective mechanism of ginsenoside Rg1 on hippocampus of aging mice induced by D-galactose. Methods Forty nestin-green fluorescent protein (GFP) transgenic mice, aged 6-8 weeks, were randomly divided into four groups: control group, ginsenoside Rg1 control group, ginsenoside Rg1 therapy group, and model group. Learning and memory abilities were measured by Morris water maze after the modeling completed. Frozen sections were made to survey the hippocampus fluorescence intensity. Senescence-associated β-galactosidase (SA-β-Gal) staining was used to detect the aging level of hippocampus. The activities of superoxide dismutase (SOD), total antioxidant capacity (T-AOC), and contents of malonaldehyde (MDA) in hippocampus were tested by chromatometry. Enzyme-linked immunosorbent assay (ELISA) was used to test the levels of interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α proinflammatory cytokins in hippocampus. The levels of p53 and p21 were detected by Western blotting. Results The learning and memory capacities of the aging model group were decreased compared with those of the drug therapy group; The fluorescence intensity in the dentat gyrus (DG) of hippocampus of the drug therapy group was increased compared with that of the model group; The SA-β-Gal positive granules in section of brain tissue of the aging model group were increased compared with those of the drug group and drug therapy group; The activitives of SOD and T-AOC of the drug therapy group were increased compared with those of the aging model group while the content of MDA was decreased. The levels of IL-1β, IL-6, and TNF-α were decreased in the drug therapy group compared with those in the aging model group. The levels of p53 and p21 were decreased in the drug therapy group compared with those in the aging model group. Conclusion Ginsenoside Rg1 can antagonistic D-galactose and delay the aging of hippocampus. In addition, improvement of anti-oxidant ability and regulation of the level of p53-p21 pathway may be the underlying anti-aging mechanism of ginsenoside Rg1.

Mechanism Study on Electroacupuncture in Reconstructing the Neurologic Function in Mice after Spinal Cord Injury / 上海针灸杂志

Objective To observe the effect of electroacupuncture at thoracic Jiaji points (EX-B 2) on the Behavior Measurement Scale (BMS) score, and expressions of nestin and glial fibrillary acidic protein (GFAP) in spinal cord in mice after spinal cord injury, and to explore the action mechanism of electroacupuncture in protecting nerves and recovering motor function.Method Ninety-six female C57BL/6 mice were randomized into a sham operation group, a spinal cord injury group, an electroacupuncture group, and an acupuncture group, 24 mice in each group. Except for the sham operation group, spinal cord injury at T9 level was induced in the other three groups. The electroacupuncture group was intervened by electroacupuncture at Jiaji points (EX-B 2) of T7 and T11, and the acupuncture group received manual acupuncture at the same acupoints. The intervention was conducted 15 min each time, once a day, with 1 d interval every 5 d, for a total of 28 d. The BMS score was evaluated respectively after 3 d, 7 d, 14 d, and 28 d intervention. Enzyme-linked immunosorbent assay (ELISA) was adopted to detect the expression ofnestin in spinal cord, and immunofluorescence and Western blotting were used to detect the expression of GFAP in spinal cord.Result Respectively 14 d and 28 d after spinal cord injury, the main and auxiliary scores of BMS in the acupuncture group were significantly higher than those in the spinal cord injury group (P<0.05); 28 d after spinal cord injury, the main and auxiliary scores of BMS in the acupuncture group were significantly higher than those in the spinal cord injury group (P<0.05) and the auxiliary BMS score in the electroacupuncture group was significantly higher than that in the acupuncture group (P<0.05). Respectively 3 d, 7 d, and 14 d after spinal cord injury, the contents of nestin in the electroacupuncture group were obviously increased and significantly different from those in the spinal cord injury group (P<0.05). The nestin content of the acupuncture group was significantly higher than that of the spinal cord injury group 7 d and 14 d after spinal cord injury (P<0.05). The expression of GFAP in the electro-acupuncture group was significantly higher than that in the spinal cord injury group 3 d and 7 d after spinal cord injury (P<0.05); the inhibitions on the immunoreactivity of GFAP in the electroacupuncture group and acupuncture group were more significant than the inhibition in the spinal cord injury group 28 d after the injury (P<0.05).Conclusion Electroacupuncture at Jiaji points (EX-B 2) can promote the recovery of motor function after spinal cord injury, which is related to the enhancement of the expressions of nestin and GFAP within 7 d after the injury, and the two expressions are in a positive correlation. In the early stage of treatment, electroacupuncture can boost the activity of astrocytes to act as neural stem cells and inhibit the immunoreactivity of GFAP in the later stage to benefit the reconstruction of neurologic function.

Relationship between Nestin expression and prognosis in human osteosarcoma / 实用医学杂志

Objective To explore the relationship between Nestin expression level and prognosis in osteosarcoma,and to provide a new idea for treatment.Methods Thirty patient with osteosarcoma who had received treatment were included this study according to the criteria.HE staining was applied to evaluate the response to chemotherapy,meanwhile immunohistochemistry was applied to evaluate Nestin expression levels before and after chemotherapy.Kaplan Meier survival curve was drawn to analyze the outcomes.Results The response of chemotherapy was good in 17 patients and poor in 13.24 cases were strongly positive in Nestin staining,five were weakly positive,and one was negative.The expression level of Nestin in osteosarcona was correlated to the chemotherapy response and prognosis (P ＜ 0.05),while higher level of Nestin expression referred to poorer response of chemotherapy and lower overall survival rate.Conclusions The expression level of Nestin in osteosarcoma tissue is associated with the effect of chemotherapy and prognosis;higher expression level of Nestin indicates poorer prognosis and efficacy of chemotherapy.

Induction of Nestin Early Expression as a Hallmark for Mesenchymal Stem Cells Expression of PDX-1 as a Pre-disposing Factor for Their Conversion into Insulin Producing Cells

Diabetes constitutes a worldwide epidemic that affects all ethnic groups. Cell therapy is one of the best alternatives of treatment, by providing an effective way to regenerate insulin-producing cells lost during the course of the disease, but many issues remain to be solved. Several groups have been working in the development of a protocol capable of differentiating Mesenchymal Stem Cells (MSCs) into physiologically sound Insulin Producing Cells (IPCs). In order to obtain a simple, fast and direct method, we propose in this manuscript the induction of MSCs to express NESTIN in a short time period (2 h), proceeded by incubation in a low glucose induced medium (24 h) and lastly by incubation in a high glucose medium. Samples from cell cultures incubated in high glucose medium from 12 to 168 h were obtained to detect the expression of INSULIN-1, INSULIN -2, PDX-1 and GLUT-2 genes. Induced cells were exposed to a glucose challenge, in order to assess the production of insulin. This method allowed us to obtain cells expressing PDX-1, which resembles a progenitor insulin-producing cell.

Nestin Expression in the Adult Mouse Retina with Pharmaceutically Induced Retinal Degeneration

The present study investigated the temporal pattern and cellular localization of nestin in the adult mouse retina with pharmaceutically induced retinal degeneration using N-methyl-N-nitrosourea (MNU). After a single intraperitoneal injection of MNU in 8-week-old C57BL/6 mice, the animals were sacrificed at 1, 3, 5, 7, and 21 days (n = 6, in each stage). The eyes were examined by means of immunohistochemical tests using nestin, ionized calcium-binding adaptor molecule (Iba-1), CD11b, F4/80, and glial fibrillary acidic protein (GFAP). Western blot analysis and manual cell counting were performed for quantification. Nestin expression was increased after MNU administration. Nestin+/Iba-1+ cells were migrated into outer nuclear layer (ONL) and peaked at day 3 post injection (PI). Nestin+/CD11b+ cells were also mainly identified in ONL at day 3 PI and peaked at day 5. Nestin+/F4/80+ cells were shown in the subretinal space and peaked at day 3 PI. Nestin+/GFAP+ cells were distinctly increased at day 1 PI and peaked at day 5 PI. The up-regulation of nestin expression after MNU administration in adult mouse retinal microglia, and monocyte/macrophage suggests that when retinal degeneration progresses, these cells may revert to a more developmentally immature state. Müller cells also showed reactive gliosis and differentiational changes.

Effects of intranasal administration of heparin-binding epidermal growth factor-like growth factor on expressions of nestin and caspase-3 in the perihematomal tissue after intracerebral hemorrhage in rats / 国际脑血管病杂志

Objective To investigate the effects of intranasal administration of heparin-binding epidermal growth factor-like growth factor (HB-EGF) on expressions of nestin and caspase-3 in the perihematomal tissue after intracerebral hemorrhage in rats. Methods A total 36 adult male Sprague-Dawley rats were randomly divided into 3 groups: an HB-EGF group, a control group, and a sham operation group (n = 12 in each group). Then they were further divided into 4-, 7-, and 10-day subgroups (n = 4 in each subgroup). A model of intracerebral hemorrhage was induced by injecting type Ⅳ colagenase. At 1 to 3 days after modeling, the HB-EGF group was intranasaly administrated HB-EGF. The control group was administrated the same volume of saline. The Bederson score was conducted at the corresponding time points and the rats were sacrificed after the forelimb placing test. Immunohistochemistry was used to detect the expression of nestin and caspase-3 in the perihematomal tissue. Results The sham operation group did not have neurological deficits. The Bederson scores (P < 0. 05) and the result of forelimb placing test (al P < 0. 01) at day 4 and 7 in the HB-EGFgroup were better than those in the control group. There were no significant differences at day 10. At day 4, 7, and 10, the number of nestin positive cels in the perihematomal tissue in the HB-EGF group was significantly more than that in the control group (al P < 0. 05), and the number of caspase-3 positive cels was less than that in the control group (al P < 0. 01). Conclusions Intranasal administration of HB-EGF can improve the early neurological function, upreguate the nestin expression and downregulate the caspase-3 expression in the perihematomal tissue after intracerebral hemorrhage in rats, indicating that intranasal administration of HB-EGF may promote the proliferation of precursor cels and decrease cel apoptosis in the perihematomal tissue.

Effect of maternal deprivation on learning and memory ability and hippocampal nerve regeneration in rats with hypoxic-ischemic brain damage / 中华实用儿科临床杂志

Objective To observe the effect of maternal deprivation(MD)on learning and memory ability and hippocampal pathology and nestin expression in rats with hypoxic -ischemic brain damage (HIBD).Methods The models of HIBD SD male rats were established by the method of Rice,and were randomly divided into 2 groups:MD group and control group.In addition,the sham -operation group(sham group)models were established.The MD group rats were separated from their mother 3 hours per day from the second day after modeling to the 21 st postnatal day.After 28 postnatal days,Morris water maze was used to evaluate the learning and memory ability of the rat models.HE stai-ning was employed to observe the hippocampal pathological change in the rats.Then,the expression of nestin in the hip-pocampus was measured by the method of immunohistochemistry.Results Their body mass changes showed that quali-ty of sham group was higher than that of the control and the MD groups,and quality was improved in the control group, compared with the MD group,and the differences were statistically significant(q =9.860 8,3.880 7,5.980 1 ,all P <0.05).The water maze scores of the MD group in place navigation test and spatial probe test were much lower than that of the control group and the sham group,and the scores of the sham group were higher than that of the control group,the differences were statistically significant(all P <0.05 ).The findings of HE stain showed that the pathology in the right -sided hippocampus of the sham group was normal and neurons were well -arranged,and that of control group was minimally abnormal,and the neurons were almost arranged orderly and remained normal.While,the pathomorpholo-gy of the MD group was obviously abnormal,the neurons were arranged disorderly,many of the neurons lost.According to the immunohistochemical findings,the number of nestin -positive cells in right -sided hippocampus of the MD group was significantly less than that of the control group,and the number of nestin -positive cells of the sham group was less than that of the MD group,which showed significant differences among the groups(all P <0.05).Conclusions MD aggravated injury to learning and memory ability of neonatal rats with HIBD,and decrease the number of nestin -posi-tive cells of MD markedly,which is not good for the recovery of brain injury.

Expression of Nestin on Endothelial Cells and Pericytes During Retinal Vascular Development in Mouse

PURPOSE: Nestin, a marker of neural stem cells, is expressed in Müller cells during retinal development. However, the role of nestin in retinal vascular development is not well established. Thus, we investigated the expression of nestin in developmental mouse retina and identified which retinal cells are related to the expression of nestin during the retinal vascular development. METHODS: Eyes were enucleated from C57BL/6 mice on postnatal day (P) 4, P8, P12, P16 and P26. Immunofluorescence was used to evaluate nestin expression in relation to endothelial cells (isolectin B4), pericytes (neural/glial antigen 2) and astrocytes (glial fibrillary acidic protein). RESULTS: Nestin was strongly expressed from the ganglion cell layer to retinoblast layer at P4. At P8, P12 and P16, the expression of nestin was observed from the upper border of the ganglion cell layer, and vertically penetrating to outer nuclear layer. At P26, the expression of nestin was decreased and confined to the ganglion cell layer and inner nuclear layer. Interestingly, there was strong vascular shape expression of nestin at all stages. The superficial, deep and intermediate vascular plexus was completely merged with nestin expression at P4, P8, P12 and P16. In addition, the nestin expression merged with pericytes but not with astrocytes. CONCLUSIONS: Nestin was expressed in endothelial cells and pericytes during retinal vascular development in the retina. These results suggest that nestin could play an important role in developmental angiogenesis via interplay with endothelial cells and pericytes.