Effects of pioglitazone on proliferation and differentiation of human preadipocytes

Objective： To explore the effects of thiazolidinediones （TZDs） pioglitazone on proliferation and differentiation of human preadipocytes. Methods：Omental adipose tissue biopsies were obtained from 15 patients who were undergoing elective open-abdominal surgery. The primary culture and differentiated induction of human preadipocytes were performed, and the human preadipo-cytes were treated with pioglitazone at different concentrations at proper moments. Dynamic morphological changes of the human preadipocytes were observed, and their proliferation and differentiation were assessed with Colorimetric MTT Assay and Oil Red O Staining. Results：After 24 hours and 72 hours with pioglitazone, 0.1 μmol/L （μmol/ml） pioglitazone increased the MTT values of the human preadipocytes by 25.3% and 34.8%,respectively（P 〈 0.05）, while 1 μmol/L pioglitazone by 27.4% and 26.6%（P 〈 0.05）, compared with the control group without pioglitazone. The human preadipocytes with pioglitazone cumulated more adipose in the endochylema than those without pioglitazone obviously. 0.1 μmol/L pioglitazone increased the differentiation degree of the human preadipocytes differentiated for 8-10 days by 44.81% and 1 μmol/L pioglitazone by 53.76%（P 〈 0.05）. Conclusion：Thi- azolidinediones pioglitazone may significantly promote the proliferation and differentiation of the human omental preadipocytes.

Effect of Acupuncture on Proliferation and Differentiation of Neural Stem Cells in Brain Tissues of Rats with Traumatic Brain Injury

Objective： To observe the effect of acupuncture on proliferation and differentiation of neural stem cells in brain tissues of rats with traumatic brain injuny. Methods： Thirty SD rats were randomly and equally allocated to the sham-operated, the model and the acupuncture groups. The traumatic brain injury model was established by the free drop method. For the rats in the acupuncture group, acupuncture was applied once a day for 7 days. Brain histotomy was carried out when treatments were completed. Immunohistochemical techniques were adopted to detect the cells that express nestin, neurofilament proteins （NF）-200 and glial fibrillary acidic proteins （GFAP）, the markers of neural stem cells, neurons, astrocytes respectively. Results： Compared to the sham-operated group, the number of nestin-positive cells and NF-200-positive cells in brain tissues was decreased significantly in the model group （P〈0.01）, whereas the number of GFAP-positive cells was significantly increased （P〈0.01）. Compared to the model group, the positive cells of nestin, NF-200, GFAP in brain tissues in the acupuncture group were increased obviously （P〈0.01）. Conclusions： Acupuncture can significantly increase the number of nestin-positive cells, NF-200-positive cells and GFAP-positive cells, indicating the significant increase of neural stem cells, neurons and astrocytes in number. Acupuncture can improve neuranagenesis by promoting the proliferation and differentiation of neural stem cells in brain tissues. This might be one of the mechanisms for acupuncture to treat traumatic brain injury and to promote the repair of nervous function.

Porcine placental peptides improve neuroblast proliferation and differentiation via enhancement of TrkB and BDNF levels in D-glatacose-induced mouse aging model

Aging affects the nervous system as well as other organs.In our study,we aimed to study the pharmacological effects and mechanism of porcine placental peptides(PPP)in aging process,and to observe the changes of neuroblast proliferation and differentiation as well as partial gene expression in hippocampus of D-galactose-induced aged mouse.Based on the analysis of experimental results,it was confirmed that PPP significantly improved neurobalst proliferation and differentiation in the mouse hippocampal DG by ki-67 and DCX immunohistochemistry.This result showed that PPP had anti-aging effects on D-galactoseinduced aging mouse model.Moreover,we observed up-regulated expressions of BDNF and TrkB proteins and down-regulated expressions of Caspase 3,8,and 9 proteins in the PPP-treated mouse hippocampus.Therefore,our results showed that PPP obviously improved neuroblast proliferation and differentiation,and its anti-aging effect might berelated to down-regulation of apoptosis-related proteins,including Caspase 3,8,and 9,via BDNF/TrkB pathway.Our findings provided valuable evidence for its applications in the health and medicine sectors.

Tissue Culture and Rapid Propagation Techniques of New Variety in Cymbidium hybridum

[Objectives]This study was conducted to select media suitable for proliferation,differentiation and rooting of Cymbidium hybridum"Huangjinjia".[Methods]The lateral buds and protocorms of the new variety C.hybridum"Huangjinjia"were used as materials to explore the effects of different concentrations of 6-BA and NAA on protocorm proliferation and rooting.[Results]The optimal medium for protocorm propagation was 1/2 MS+6-BA 1.0 mg/L+NAA0.5 mg/L+potato 50 g/L+sucrose 20 g/L,in which the protocorms multiplied easily and grew rapidly.The optimal medium for inducing adventitious buds was1/2 MS+6-BA 1.5 mg/L+NAA 0.3 mg/L+sucrose 30 g/L+banana 25 g/L+apple 25 g/L+activated carbon 1.0 g/L,in which the induction rate of adventitious buds reached 335%.The optimal medium for rooting was 1/2 MS+NAA 0.5 mg/L+sucrose 25 g/L+banana 75 g/L+apple 25 g/L+activated carbon1.0 g/L,in which the average root length was 3.0 cm,the average number of roots was 2.6,and plantlets had green leaves,thick roots and suitable plant height.[Conclusions]This study provides a theoretical basis and reference for the establishment of a rapid propagation system using lateral buds.

Effects of Rat Cytomegalovirus on the Nervous System of the Early Rat Embryo

The purpose of the study was to investigate the impact of rat cytomegalovirus （RCMV） infection on the development of the nervous system in rat embryos, and to evaluate the involvement of Wnt signaling pathway key molecules and the downstream gene neurogenin 1 （Ngnl） in RCMV infected neural stem cells （NSCs）. Infection and control groups were established, each containing 20 pregnant Wistar rats. Rats in the infection group were inoculated with RCMV by intraperitoneal injection on the first day of pregnancy. Rat E20 embryos were taken to evaluate the teratogenic rate. NSCs were isolated from El3 embryos, and maintained in vitro. We found： 1） Poor fetal development was found in the infection group with low survival and high malformation rates. 2） The proliferation and differentiation of NSCs were affected. In the infection group, NSCs proliferated more slowly and had a lower neurosphere formation rate than the control. The differentiation ratio from NSCs to neurons and glial cells was significantly different from that of the control, showed by immunofluorescenee staining. 3） Ngnl mRNA expression and the nuclear p-catenin protein level were significantly lower than the control on day 2 when NSCs differentiated. 4） The Morris water maze test was performed on 4-week pups, and the infected rats were found worse in learning and memory ability. In a summary, RCMV infection caused abnormalities in the rat embryonic nervous system, significantly inhibited NSC proliferation and differentiation, and inhibited the expression of key molecules in the Wnt/β-catenin signaling pathway so as to affect NSCs differentiation. This may be an important mechanism by which RCMV causes embryonic nervous system abnormalities.

Effect of Porous Activated Charcoal Reinforcement on Mechanical and In-Vitro Biological Properties of Polyvinyl Alcohol Composite Scaffolds

The present work focused on developing an innovative composite material by reinforcing polymer matrix with highly porous activated charcoal. Polyvinyl alcohol-activated charcoal（PVA-AC） composite scaffolds were developed by varying the AC concentrations（0, 0.5, 1, 1.5, 2 and 2.5 wt%） in PVA matrix by freeze drying method. The developed scaffolds were characterized for their physicochemical, mechanical and in-vitro biological properties. In addition, the effect of AC on the attachment, proliferation and differentiation of osteoblast MG 63 cells was evaluated by scanning electron microscopy（SEM）, 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide（MTT） assay, alkaline phosphatase（ALP） activity assay and alizarin red stain-based（ARS） assay. All the PVA-AC composite scaffolds exhibited good bioactivity, hemocompatibility and protein adsorption properties. The scaffolds with high AC concentration（2.5 wt%） showed controlled drug release kinetics that are suitable for long term healing. The mechanical properties of all the PVA-AC composite scaffolds were improved when compared to the pure PVA scaffold. The high porosity, swelling degree and hydrophilicity of PVA-AC composite scaffolds facilitated cell attachment and proliferation. This is due to porous AC present in the sample that supported the osteoblast differentiation and formed mineralized nodules without the addition of any extra agents. From the above studies, it can be concluded that PVA-AC composite scaffolds are promising biomaterials for bone tissue engineering applications.