Objective To research on the proliferation and differentiation in vitro of primordial germ cells. Methods We got plenty of primordial germ cells(PGCs) by co-culture of PGCs with sertoli cells(SCs).The primary cells formed embryoid bodies(EBs) by suspension culture.furthermore,these EBs were plated on gelatin-coated culture plates for spontaneous differentiation. Results We got plenty of PGCs by coculture.Primary PGCs and passaged PGCs for histochemistry staining with AKP and immunohistochemistry staining for SSEA-1 display positive character.The EBs by suspension culture differentiated into epithelial-like,fibroblast-like and neuronal-like cells in vitro. Conclusion Primordial germ cells can proliferate and keep the character of stem cells by co-culture with SCs.PGCs can form EBs by suspension culture,most of these EBs can differentiate into neuronal-like cells,they also can differentiate into epithelial-like and fibroblast-like cells in vitro.

Objective To measure and compare sevoflurane minimum alveolar concentration (MAC)in newborn and adult rats.Methods The rats were divided into newborn rat (6-8 days) group (25 cases)and adult rat (8-10 weeks)group (25 cases).All rats were settled in the self-made device for anesthesia with inhaled sevoflurane,and the tails of rats were exposed out of the device. Sevoflurane was given via an anesthesia machine.Up-and-down method and clamping tail stimulus were applied to measure the MAC values of sevoflurane in the two groups.1.5% of sevoflurane was set as the initial concentration,and± 0.2% as adjustable gradient.A positive or negative response was judged by clamping tail stimulus in a 20 minutes interval.The mean MAC value of all rats in each group was defined as the MAC value.Results The MAC value of newborn rats was (2.58 ± 0.1 1)%,and the MAC value of adult rats was (2.32 ±0.13)%.A significant difference was found between the two groups (P <0.01).Conclusion The MAC value of newborn rats was much higher than that of adult rats.Newborn rats need a higher concentration of sevoflurane at the same depth of anesthesia when compared with adult rats.

Purpose:Home-made NiTi-stent was used in the study to observe the biological compatibility.Materials card Methods:Stents were implanted within the bile ducts,arteries,veins,bronchi of swine by surgery.Swine was killed according to the tie schedule to observe the patency of stents.Results:6 of 11 biliary stents were occluded completely,with only 5 stents partially patent.The degree of occlusion was related to the time period.10 of 11 femoral stents were patent,only 1 stent was occluded completely.1 of the only 1 venous stent was occluded completely,4 of 8 bronchial stants were patent.Epithlium tissues were found along the surfaces of stents.Both sides of the stents were covered by the epithelium cells partially and dominated by the proliferation of connective tissues and parenchymal cells of the organs.The constitution of proliferated tissues was related to the implanted time period of the stents.No connective tissues were found within two weeks,but obvious proliferation of connective tissues were found associated with lympheytic tissues.Conclusion:The dagree of surface covering of the home- made stent by epithelial the rate of was related to the diameters of the of and the lumceh stent implantation segment also the flow volume within the stent,and the period of stent implantation.Good results can be yielded by selecting the suitable stent and the implanting site.

Analyse ethic problem in hospital infection management process through discussing cause and effect of Infection and put forward to resolve approach and countermeasure.

Objective To investigate the therapeutic efficacy of the canalith repositioning maneuver to benign paroxysmal positional vertigo (BPPV). Methods Epley maneuver, Barbecue rotation and Semont maneuver were applied to twelve cases of BPPV. Results Positional vertigo in all subjects disappeared completely 48 hours later by treatment with manipulative reduction, and no obvious adverse reaction was found. There was no recurrence during a follow up from 3 to 11 months.Conclusion The canalith repositioning maneuver is effective, simple and safe for the patients with BPPV and may be recommended as the first-selected treatment modality.

Objective To study the relationship between hyperhomocysteic acid(HHcy) and artherosclerosis.Methods 20 male rabbits were randomly divided into control and experiment groups(n=10).HHcy models were made by hypodermical injection of DL-methionine for 7 weeks.After 7 weeks homocysteic acid(Hcy),IL-1?,IL-6,and IL-8 in serum of rabbits in two groups were measured by enzyme linked immunosorbent assay(ELISA).And the pathomorphological changes of aorta were observed with HE staining.The number of NF-?B positive cells in the aorta were counted by immunohistochemical method.Results The levels of Hcy,IL-1?,IL-6 and IL-8 in experiment group were higher than those in control group(P

Objective To describe the disease's spectrum of fever patients,especially to analyze the epidemic characterization of infectious diseases,to offer the proof for fever department in screening of infectious diseases.Methods A total of 15081 patients were analyed for patients presenting to the Infectious Diseases Department,People's Hospital,Peking University,between June 2005 to May 2006.A retrospective study was used to analyze the disease's spectrum and epidemic characterization.Results Among the disease's spectrum of fever patients,the first leading cause was respiratory diseases(78.22%),followed by infection with the special patients(10.50%)、acute gastroenteritis(3.57%)、the infection at other site with etiological factor determined(2.82%)、infectious diseases(2.30%)、urinary system infection(2.06%) and fever of unknown origin(0.53%),respectively;Among the spectrum of infectious diseases,the first leading cause was infectious diarrheal disease(29.97%),followed by measles(24.78%)、lung tuberculosis(11.24%).Conclusion The spectrum of fever is complex.To obtain the epidemic characterization of infectious diseases,it's helpful to the fever department to screen and prevent the infectious diseases.

The patients with cor pulmonale in release stage have been treated and cared with many methods.The results showed that there are obvious recuperatio of respiratory symptoms and lung function and evident strength of the immune function of the body and movement of diaphragm in the patients of treatment group with cor pulmonale compared with control group.The studies suggested that the rehabilitation treatment and care plays an important role in treating patients with cor pulmonale,particularly postponing and prevention acute attack.

BACKGROUND:In vitro isolation and purity technique of stem cells mostly depends on the identification of cell surface marker,such as monoclonal antibody adherent spreading method,flow cell sorting method and immunomagnetic beads sorting method,but the operation was complicated and the price was high.OBJECTIVE:To observe the biological characteristics of human amniotic fluid-derived embryonic mesenchymal stem cells,which were isolated and cultured using the two-step method.DESIGN,TIME AND SETTING:The opening study was conducted at the Stem Cell Research Room of Xinjiang Medical University from March 2008 to March 2009.MATERIALS:Totally 10 amniotic fluid specimens were obtained from pregnant women who underwent prenatal diagnosis following 16-22 weeks of gestation or voluntarily induced abortion.With ultrasonic guidance,amniocentesis was performed to collect 20-40 mL amniotic fluid.METHODS:Human amniotic fluid-derived embryonic mesenchymal stem cells were isolated and cultured using the two-step method.Amniotic fluid was first centrifuged and incubated till spindle-shape cells were seen,with the presence of flbroblast-tike cell colonies.Supematant was moved to a new 25 cm~2 culture flask for further culture till spindle-shape fibroblast-like mesenchymal stem cell colonies.When 70% confluence,cells were digested,and incubated in α-MEM,supplemented with basic fibroblast growth factor,served as the first passage.MAIN OUTCOME MEASURES:Morphological changes in human amniotic fluid-derived embryonic mesenchymal stem cells of primary culture and subculture were measured.Karyotype,cycle,growth curve and colony formation ability of human amniotic fluid-derived embryonic mesenchymal stem cells were measured.Surface antigen and cytokine were examined using flow cytometry,immunofluorescence and RT-PCR.RESULTS:Human amniotic fluid-derived embryonic mesenchymal stem cells were successfully isolated and subcultured.During metaphase,primarily cultured amniotic fluid cells presented scattered spindle cells and flbroblast-like mesenchymal stem cell colonies every 7 days.Passaged cells completely adhered in 12 hours.Following 1 or 2 days of latent period,cells proliferated rapidly.About 90% confluence was observed following 6 or 7 days of culture.Cell arranged regularly,showing whirlpool-shape,radiated shape.Cells were spindle-shape,with unclear boundary.Chromosome karyotype of human amniotic fluid-derived embryonic mesenchymal stem cells was normal diploid.Growth curve showed "S" shape,but the two-step method reached a peak at (6.1±0.5) days,which was significantly rapid compared with the one-step method (7.2±0.6) days (P=0.035).Flow cytometry analyses showed that P3 cells at S phase took up (14±2.3)% using the two-step method,which was more than the one-step method (9.0±1.4)% (P=0.031).Low-density human amniotic fluid-derived embryonic mesenchymal stem cells were incubated for 7 days prior to cells formed scattered cell colonies.However,colony forming efficiency using the two-step method (15.0±2.3)% were significantly more than the one-step method (10.0±1.8)% (P=0.021).Flow cytometry results showed that human amniotic fluid-derived embryonic mesenchymal stem cells expressed CD44,CD29 and CD105,but were negatively for CD45,CD34,HLA-DR.Immunofluorescence suggested that Oct-4-positive cells were observed in amniotic fluid.However,the proportion of Oct-4-positive cells using two-step method (1.2±0.3)% was significantly greater than the one-step method (0.9±0.2)% (P=0.041).RT-PCR suggested that human amniotic fluid-derived embryonic mesenchymal stem cells obtained using the two methods expressed Oct-4.CONCLUSION:Human multipotent mesenchymal stem cells are present in human amniotic fluid.The two-step culture protocol could be a kind of high performance and simple protocol which may not interfere with the normal prenatal diagnosis procedure.

ObjectiveTo explore the effects of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/nuclear factor-kappa B (NF-κB) signal pathway on the process of follicle-stimulating hormone (FSH) facilitating cell proliferation and invasion in human epithelial ovarian cancer. Methods Ovarian cancer cell lines SKOV3 and 3AO were cultured to exponential phase,then assigned to control group,FSH group,LY294002 group and FSH + LY294002 group,respectively.Cells were treated with different concentration of FSH and LY294002,respectively.The effects of FSH on cell proliferation were observed by methylthiazolyl tetrazolium (MTT).Morphological changes were observed by phase contrast microscope.The ability of cell invasion was investigated by transwell invasion assay.The expression of FSH receptor (FSHR),Akt1/2,phosphorylated-Akt (p-Akt) and NF-κB p65 protein were detected by western blot.Results( 1 ) FSH could promote the proliferation of SKOV3 and 3AO cells.When the cells were treated with 40 U/L FSH for 48 hours (SKOV3) and 24 hours (3AO),compared with those in control groups,they reached the highest proliferation rate (P ＜ 0.05 ),respectively.(2) The morphology of SKOV3 and 3AO cells in four groups:in control group,SKOV3 cells were short spindle and 3AO cells were long spindle,the nuclei of them were both roundness or oval,the cytoplasm were bright.In FSH group,the cells changed to slightly longer or polygonal,they were full in shape,meanwhile,the cell intensity were higher than control group.In LY294002 group,some cells changed from spindle to round,and began to shrink.The cell intensity diminished.The morphology of FSH + LY294002 group was similar with control group,but the cell intensity was lower than that in FSH group.(3)The number of SKOV3 cell that passed through the membrane in control group,FSH group,LY294002 group and FSH + LY294002 group was (26 ± 6),( 118 ± 19),( 18 ± 5) and ( 38 ± 7 ),respectively.The number of 3AO cell was ( 19 ± 4 ),( 134 ± 20),(12 ±3) and (58 ± 11 ),respectively.The results showed that the number of cells in FSH group was significantly higher than that in control group ( P ＜ 0.05 ),while the number of cell in FSH + LY294002 group was significantly fewer than that in FSH group (P ＜ 0.05 ).(4) There was no significant difference in the expression of FSHR and Akt1/2 between FSH group and control group (P ＞ 0.05 ),but FSH increased the expression of p-Akt and the ratio of NF-κB p65 in the nucleus versus cytoplasm in SKOV3 and 3AO cells,there were significant differences compared with control group ( P ＜ 0.05 ).LY294002 reversed the effects of FSH on increasing the expression of p-Akt and the ratio of NF-κB p65 in the nucleus versus cytoplasm,there were significant differences among LY294002 group,FSH + LY294002 group and FSH group (P ＜ 0.05 ).ConclusionThe effects of FSH on proliferation and invasion of ovarian cancer cell lines SKOV3 and 3AO may be realized by regulating the activity of NF-κB in PI3K/Akt signal pathway.