Objective To obtain eukaryotic expression vectors containing coding region of the gene BC203882 and detect its effect on cell growth.Methods The coding region of the gene BC023882 was amplified by RT-PCR,then cloned into eukaryotic expression vector pcDNA3.1(-)/Myc-His(+)/lacZ.The recombinant plasmid was transfected into embryonic fibroblast of mouse and COS-7 cells with liposome method.Its effects on cell proliferation were examined by FACS and growth curves.Results The eukaryotic expression vectors containing coding region of the gene BC203882 were constructed.The growth of the transfected cells was retarded,and the proliferation index of cells was significantly decreased.Conclusion The eukaryotic expression vectors of the gene BC023882 were constructed successfully and the inhibitory effect of the gene on cell growth was observed.

Objective To clone and identify neurotrophic factor NT3 gene from mouse liver so as to establish a cell strain expressing high level of NT3 gene. Methods The target genes amplified by RT PCR were cloned into the shuttle vector pExchange 1neo, and then the DNA sequence was analyzed by enzyme digestion and sequencing. The recombinant expression vector pExchange NT3 1neo was employed to transfect the embryonic stem cell strain MESPU35, and then the transfected cells were selected by G418. The NT3 level in the transfected cells was detected by RT PCR. Results A gene fragment of 777 bp was obtained by RT PCR, and the DNA sequence was identical to mice NT 3 gene sequence of GenBank. The recombinant vector was constructed successfully and the constructed cell strain could express high level of NT 3 gene. Conclusion The successful cloning of NT3 gene from mouse liver, construction of pExchange NT3 1neo expression vector, and establishment of cell strain stably expressing high level of NT3 gene have laid a foundation for the further studies.

BACKGROUND: Neural axon regeneration is one of the difficulties that must be overcome in treatment of injury of central nerve system. Significant therapeutic effects have been obtained in transplantation of neural stem cells (NSCs), embryonic stem cells (ESCs) and Schwann cells. But the bottleneck situation of insufficiency of cell provider has limited the development on it.OBJECTIVE: To observe directional-differentiation of retinoic-acid induced ESCs so as to find optimal condition for neuronal differentiation.DESIGN: Non-randomized controlled experiment was designed.SETTING: Teaching-Research Room of Histology and Embryology, Department of Basic Medicines. Third Military Medical University of Chinese PLAMATERIALS: The experiment was performed in Staff Room of Histology and Embryology, Third Military Medical University of Chinese PLA from January to May 2000. Eighteen Kunming mice in disoestrus were employed, of which. 12 mice were female and 6 mice male. They were placed in same cage at ratio of 2:1 for mating. The date of pregnancy was recorded. MESPU35 ESC line was prepared.METHODS: Removed head. internal organs and four limbs, feeder-layer Feeder-layer adherent culture was used to proliferate MESPU35 ESCs.Classic 4-/4+ method [The embryoid body (EB) grew naturally for 4 days,without retinoic acid added. In the coming 4 days, retinoic acid was added to induce neural EB of high proportion] was applied to induce the directional differentiation of the nerve. EB was cultured with serum of different concentrations. Phase contrast microscope was used to observe nerve-like EB in serum of different concentrations and to count numbers. ②Immunocytochemical technique was used to observe cellular morphological charac ters at various differentiating phase spots (5th. 9th, 14th days) and with retinoic acid at various concentrations. Flow cytometer (FCM) was used to count the proportion of differentiated neurons.MAIN OUTCOME MEASURES: ①Estimated measurement of the length of process and cell body during formation of neural EB after retinoic-acid induced differentiation of MESPU35 ESCs. ② Observation of cell morphology with immunocytochemical staining and proportion of differentiated cells assayed with FCM.RESULTS: ①It was discovered with phase contrast microscope that serum of different concentrations affect neural directional differentiation after EB formation to certain extent. Excessively high and low concentrations of serum reduced the proportion of neural differentiation of EB. The differentiating proportion is high in serum with 5% concentration. ② It is observed with immunocytochemical technique that the proportions of NF200 positive cell and glial fibrillary acidicprotein (GFAP) positive cell in differentiation of MESPU35 ESCs induced by retinoic acid were increased with phase spots in differentiation and increased concentration of retinoic acid. NF200 positive cell is transformed as multipolar neurons from absence of process in morphology. The processes of GFAP positive cell became longer and linked among each other as reticular pattern finally. ③ It was assayed with FCM that the proportion changes of GFAP positive cell and NF200 positive cell manufactured in differentiation were similar to immunocytochemical one.CONCLUSION: Retinoic acid in combination with proper concentration of serum and differentiating phase spots can induce neural-differentiation of MESPU35 ESC at high proportion and its differentiating regulation is in the patterns of concentration dependence and time dependence.

Objective To establish human neural stem cells(HNSCs) model for further basic research and clinical application. Methods Cells from human fetal cerebral cortex were collected and cultured with serum free midium and then identified for nestin immunocytochemical staning;The cells were induced to differentiate by 5% fetal bovin serum and identified by neurofilament-200(NF-200) and glial fibrillary acidic protein(GFAP) immunocytochemical staning. Results The harvested cells appeared as clusters in suspension and both NF-200 and GFAP positive cells were observed after induction.After 12 generations of culture,these cells retained the main characteristics of NSCs.Conclusions The HNSCs were harvested from human fetal cerebral cortex and this HNSCs model can be used for futher basic research and clinical applications.;

It is important and necessary for the teaching process in histology and embryology integrated by psychological quality.The psychological quality of teachers can be improved by professional training and by themselves.Teachers should teach everyone differently according to the different psychological character of medical students who are born after 1990.Teachers can improve psychological quality of medical students in teaching process including the discussion,visiting the embryo sample and the second class.

Pender′s health promotion model explains the factors influencing health behaviors, which provides a framework for nursing practice and research. Functional exercise compliance in postoperative patients with breast cancer was in a low level, this article reviewed factors influencing functional exercise among breast cancer survivors through three aspects based on health promotion model and made some suggestions on nursing intervention, to promote the rehabilitation of this population.

Objective To detect the expression changes and location of the mouse developmental regulation brain protein(Dbn1)in the developmental mouse brain.Methods Monoclonal antibodies against drebrin protein were used to assess Dbn1 by immunohistochemistry and Western blot.The paternal expression of Dbn1 in developmental mouse brain(E14,P1,P7 and adult)was initially investigated by Western blot.Dbn1 was shown at various developmental stages(E14,P1 and P7)as well as in adult in different brain area of developmental mouse brain by immunohistochemical.Results Dbn1 protein was detected in developmental brain but a little in adult brain by Western blot,high at E14,decreased at P1,gradually increased at P7 and lowest in adult.Immunohistochemistry confirmed as follows:Dbn1 expressed mostly in cortex,hippocampus and ependyma areas at E14,and the positive signal was distributed at cells border;The expression of Dbn1 was decreased at P1,mainly distributed at the verge of cells or dendrites;The peak expression of Dbn1 appeared at P7,Dbn1 located at nucleus of hippocampus and cortex,and the positive signal located within cytoplasm and dendrites;Only a little positive Dbn1 cells were found in adult mouse brain.Conclusion Dnb1 may be involved in regulating the differentiation and migration of neurons during the development of mouse brain.

Objective To isolate and identify pancreatic stem cells of Kunming mouse and to observe the differentiation potency of those cells. Methods Pancreas of postnatal Kunming mice were digested by enzymes to isolate pancreatic stem cells. The potency of the cell differentiation was then identified with special marker of cytokeratin 19(CK 19), insulin and glucagons by immunocytochemical staining. Results Few epithelioid cells could be found to survive at the beginning of isolation, but when medium was replaced by that with growth factor, the cells proliferated quickly in fusiform shape and formed colony gradually. The cells were CK 19 immunoreactive positive after transfer of culture. After differentiation induced by high glucose, the cells formed the pancreatic islet like structures. Immunocytochemical staining showed that part of the cells of pancreatic islet like structures were insulin immunoreactive positive and few of the cells were glucagon immunoreactive positive. Conclusion Pancreatic stem cells of Kumming mouse can proliferate when cultured in vitro and have the potency of differentiation into ? and? cells of pancreatic islet.

Objective To investigate the safe and effective method for laryngeal microsurgery in difficult la‐ryngeal exposure cases .Methods We selected 62 patients’ clinical data who had received laryngeal microsurgery with difficult laryngeal exposure and could not exposure by normal self -retaining laryngoscope between July 2012 and June 2015 .There were 42 cases of vocal cord polyp ,9 cases of the vocal cyst ,5 cases of the vocal amyloidosis , 4 cases of severe atypical hyperplasia of vocal cords and 2 cases of vocal cord high differentiated squamous carcino‐ma .We completed all kinds of laryngeal microsurgery to expose the glottis by adjusting the postures of patients ,in‐creasing the anesthesia depth ,using self -retaining laryngoscope with endoscopy which can be adjusted and pressing the throat .Results In 62 patients ,58 patients were successfully operated with adjustable self -retaining laryngo‐scope with endoscopy ,the success rate was 93 .55% .And 25 cases was exposed the glottis completely by increasing the anesthesia depth ,however ,when we increased the anesthesia depth ,there were 10 cases needed to combined with pressing the throat to expose .Five patients had retropharyngeal injure with different levels .One case with small jaw deformity of the vocal cord polyp surgery was not successful ,the success of electronic endoscopic under surface anesthesia surgery .The other one case with teeth unkempt and porcelain teeth and two cases of intraoperative frozen tip vocal cord cancer completed the operation of the open throat under the non trachea incision .Conclusion Most of difficult exposed laryngeal can be safely and effectively exposed through using the adjustable self -retaining laryngo‐scope with endoscopy while normal self -retaining laryngoscope can not .When necessary ,we can put 30°endoscope into the side channel of self -retaining laryngoscope to complete all kinds of laryngeal microsurgery .