Objective To evaluate the effects of IL-1? and combination of IL-1?,IL-11,LIF and GDNF on inducing human neural stem cells(hNSCs)to differentiate into dopaminergic(DA)neurons in vitro. Methods A great deal of neurospheres was obtained by the technology of serum-free culturing and single-cell cloning,and was splitted into 3 groups,which were cultured in different media.In control group,the differentiation medium used only contained 10%FBS.In IL-1? group,the medium contained IL-1? and 10%FBS.In united factors group,the medium contained IL-1?,10%FBS supplemental with IL-11,LIF and GDNF.After 3 weeks,the tyrosine hydroxylase(TH)positive cells were detected by using immunofluorescence,and image processing about the number,the induced differentiational rate,the cell bodies' areas and the cells' perimeters of TH positive neurons was carried out.TH/MAP-2 double-immunofluorescence labeling was used to calculate the percentage of DA neurons in total neurons. Results In the control group,there were few TH positive neurons with poorly developed morphology.The presence of IL-1? induced more DA neurons,but these cells were still immature.In the united factors group,the number of maturer TH positive DA neurons was the most. Conclusion IL-1? has an obvious effect on inducing hNSCs derived from human fetal mescenphalon to DA neurons.The utilization of IL-1?,IL-11,LIF and GDNF in combination has a cooperative effect on inducing differentiation of hNSCs to mature DA neurons.

Objective To investigate the difference of Brn-4 mRNA expression between the fimbria/fornix-transected rats and normal ones. Methods Forty-two SD rats were randomly divided into 7 groups,6 rats in each group.One group served as normal control and the others served as fimbria/fornix transected 1st,3rd,7th,14th,21st and 28th day group,respectively.Then hippocampi were isolated and total RNA was extracted.Semi-quantitative reverse transcriptase-polymerase chain reaction(RT-PCR) method was used in detection of the expression change of Brn-4 mRNA in hippocampus after fimbria/fornix transection.Results The expression level of Brn-4 mRNA started to increase on the 3 days after fimbria/fornix transection.The peak appeared on the 14 days,then decreased slowly to pre-transection level on the 28 days.Conclusion The process that the expression of Brn-4 mRNA increases significantly after fimbria/fornix transection,perhaps,might be related to the transplanted neural stem cells(NSCs) differentiating into neurons and AChE positive neurons in hippocampus.

Objective To study the influence of HBV infection for gastromucous membrane in chronic hepatitis B and cirrhosis and its pathogenesis.Methods The liver biopsy,gastroscope,liver function and HBVM were detected at the same time in 60 patients with chronic hepatitis B and cirrhosis.HBsAg and HBcAg in the tissues of liver and gastromucous membrane were detected by SP immunohistochemistry.Results The damage of gastromucous membrane were found in 56 of 60 patients.Its damage degree was showed as moderate and severe.The other 34 showed mild and moderate damage(P

Objective To research a new algorithm to get the relative pose parameters of spacecraft during the final approach phase of spacecraft rendezvous and docking(RVD).Methods The method was based on the shape of five non-coplanar points and the model of the pinhole,and the conception of 3D reconstruction was employed.For the shape of the mark was known,the 3-D shape of the cross-mark was reconstructed based on the single visual system,and the closed-form algorithm for relative pose parameters of spacecraft was given.Results The algorithm had been validated by the mathematical simulation,and the precise results of the relative pose parameters were calculated.Conclusion The simulation results show that the algorithm is rather simple and with relative accuracy.Furthermore it is meets exact the need in real time.

Objective To study the effect of glycerin fructose combined with mannitol in the treatment of postoperative swelling of extremities fractures.Methods In Yiwu Central Hospital from February 2015 to February 2017 treated 100 cases of limb fracture surgery patients as the research object in the course of the study, were randomly divided into control group and experimental group two were 50 cases each.The patients in control group were treated with mannitol after fracture surgery, and the patients in the experimental group were treated with glycerol fructose and mannitol.Comparative analysis of the experimental group and control group of patients with limb swelling degree.Results After different treatment, the experimental group of patients with adverse reactions occurred in 0 cases, the probability was 0.0%, the control group of patients with adverse reactions in the number of cases in the case of the number of patients with adverse reaction probability of 10.0%, the number of patients with adverse reactions in the control group was 5.As a result, the incidence of adverse reactions in the experimental group was significantly lower than that in the control group, with statistical difference(P<0.05).In the experimental group, 6 days after surgery, on the 7 day, the 8 day and the 9 day, the forearm circumference was significantly lower than that of the control group, with statistical difference(P<0.05).On the 8 day after surgery and after 8 days, the leg circumference of the experimental group was significantly lower than that of the control group, with statistical difference(P<0.05).Conclusion The clinical effect of glycerol and fructose combined with mannitol in treatment of limb swelling after surgery for fracture, high safety, reduce the probability of occurrence of adverse reactions to a great extent, is further applied in clinical significance.

BACKGROUND:Clinical application of neural stem cells is under exploration.Currently,the indicative differentiation of neural stem cells into specific neurons to replace lost and degenerative neurons needs to solve.OBJECTIVE:To explore the effect of 83 ku protein in rat hippocempi on inducing neural stem cell differentiation into acetylcholine esterase (ACHE) positive neurons.DESIGN,TIME AND SETTING:In vitro controlled observation of cytology was performed at the Medical College of Nantong University between October 2003 and April 2008.MATERIALS:A total of 12 SD rats,of clean grade,and SD fetal rats,aged 17 days,were provided by the Experimental Animal Center of Nantong University.METHODS:The normal hippocampi and hippocampi on the 14th day after the hippocampal fimbria transection were prepared into homogenate used for 10% native-polyacrylamide gel electrophoresis,and the differential proteins of 83 ku were electroeluted.The protein concentration was adjusted to 300 mg/L.The forebrain tissues of fetal rats were harvested and neural stem cells were isolated and in vitro cultured:blank control cells were cultured in serum-free DMEM/F12 medium;83 ku normal and 83 ku transection groups were separately cultured in serum-free DMEM/F12 medium containing 10 mg/L 83 ku protein from normal and hippocampal fimbria transection rats for 12 days.MAIN OUTCOME MEASURES:AChE histochemical staining was used to detect the differentiation of neural stem cells into AChE positive neurons.RESULTS:After 12 days of culture,there was a large amount of AChE positive neurons in 83ku transection group and their bodies were very big and the processes were abundant;The AChE positive neurons in 83ku normal group were less than 83 transection group,and their bodies were small with short processes.A few of AChE positive neurons were seen in control group.There were significant differences in number of AChE positive neurons among three groups (P＜0.05).CONCLUSIONHippocampal 83 ku protein can induce neural stem cells to differentiate into AChE positive neurons.

Objective To investigate the effect of extracellular signal-regulated kinases(ERK) signal transduction in the process of NSCs differentiating into neurons in the fimbria-transected hippocampi's extracts. Methods Twelve Sprague-Dawley rats'right fimbrias were transected. The extracts were gained from the fimbria-transected hippocampi at the 14th day normal rat, and the extracts supernatant fluid was collected after centrifugal process, then the protein concentration in the extracts was determined. In the serum-free medium,NSCs from the fetal hippocampus were planted on 24 well culture plate, then were divided into three group and eight wells for each group as follows: the transected group contained the extracts of the fimbria-transected hippocampi;the normal group contained the extracts of the normal hippocampi;the pure control group have no extracts. After cultured for 14 days,the cells were detected by using MAP-2 and p-ERK immunofluorescence. Result The number, area, perimeter of MAP-2 positive neurons were all declined in transected group, the normal group and the control group orderly. Statistic results showed significant difference between every two groups. The number of MAP-2/p-ERK double-positive neurons were decreased in transected group, the normal group and the control group orderly, but the percentage of double-labeled neurons in total MAP-2 positive neurons were increased in turn. In these two aspect, there were also significant difference between every two group. And most of the MAP-2/p-ERK double-positive neurons were immature. Conclusion The extracts of the fimbria-transected hippocampi had obvious effects on promoting NSCs differentiating into neurons and speeding up the maturation of neurons than those of the normal hippocampi. The morphological results showed that ERK signal transduction might be related to the differentiation of NSCs into neurons.

Objective To investigate the relationship between the nerve growth factor ( NGF ) induced hippocampal neuroregeneration and homeobox gene Lhx 8.Methods Seventy-two SD rats were divided into control group , transected group, NGF group, transected combined with NGF group after right fimbria-fornix transection and NGF intracerebroventricular injection . Real-time PCR and Western blotting were applied to detect the gene and protein expression of Lhx8 in each group.The choline acetyltransferase ( ChAT)/Lhx8 double labeled cells in subgranular zone ( SGZ) of hippocampus in each group were detected by immunofluorescence .Results The expression of Lhx8 gene and protein in the transected , NGF group and especially in the transected combined with NGF group was obviously higher than in the control group .The number of ChAT/Lhx8 double labeled cells in the NGF group and the transected combined with NGF group was obviously more than in the control group and transected group . Conclusion The hippocampal neuroregeneration which induced by NGF intracerebroventricular injection was associated with the higher expression of Lhx8.

Objective Modified the medium that can increase single\|clone formed rate and confirmed the single clone spheres had the multipotential of differentation. Methods We modified the medium, that is, the medium contained half of primary culture medium and half of fresh culture medium. A great deal of neurospheres dervied from a single cell were plated averagely into 24 well plates and added into the DMEM differentiation medium (containing serum). After culturing for 14 days, cultures were stained with the neuronal\| ang glial\|specific markers (MAP\|2 for neurons, GFAP for astrocytes and CNP for oligodendrocytes). Results Each 96 well plate containing half of primary culture medium generated two to three single clone spheres, in control plate containing only fresh medium generated half to one single clone sphere. After differentiation, these cell clones expressed MAP\|2, GFAP and CNP positive respectively.Conclusion\ Using half of primary culture medium can increase single\|clone formed rate and these cell clones had the multipotential of differentiation.\;[

Objective To evaluate the effects of ginkgolide B on inducing neural stem cells differentiating into neurons. Methods A great deal of single cell clone neurospheres raised from single cell and proliferated by the technology of serum\|free culture and single cell clone.Suspensions of cell clone neurospheres were plated equably into 24 well plates and added into the 10% FBS differentiation medium containing ginkgolide B,BDNF or without any factor.Cultures were terminated at 7 and 14 days respectively.MAP\|2,neuron\|specific marker,were used to mark neurons by immunofluorescence.MAP\|2 positive neurons were observed and counted by fluorescence microscope.The area and perimeter of these positive neurons were analyzed. Results The number of MAP\|2 positive neurons in ginkgolide B group is more than that in the control group in two periods.The area and perimeter of MAP\|2 positive neurons in ginkgolide B group were markedly larger than those in the control group at 7 and 14 days after cultured,but it's less than those in the BDNF group.Conclusion\ Neural stem cells can be induced to differentiate into neurons by ginkgolide B which has the similar role to BDNF.\;[