Air Pollution, Indoor ; analysis ; Environment, Controlled ; Humidity ; Radon ; analysis ; Temperature

BACKGROUND: The neural stem cells (NSCs) will be clinically applied extensively in the future, and seeking of more promoting methods of the proliferation and differentiation of NSCs in vitro will be the study direction of NSCs.OBJECTIVE: To introduce an economic and time-saving method for NSCs proliferation.DESIGN: Observation and control experiment.SETTING: Beijing Neurosurgical Institute.MATERIALS: Four Wistar rats pregnant for 14 days with the body mass of (180±20) g (bought from Animal Department of Chinese Academyof Medical Sciences with the batch number of SCXK1100-0006 for experiment animals) were selected and fed at common temperature and humidity.The DMEM/F12 and B27 were bought from Gibco Corporation. The basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) were bought from PeproTech Company. The nidogen monoclonal antibody (MA),glial fibrillary acidic protein (GFAP) polyclonal antibody, galactocerebroside polyclonal antibody and microtubule-associated protein 2 (MAP2) were obtained from Chemicon Company. The fetal bovine serum (FCS) was provided by Hyclone company.METHODS: The experiment was conducted in the Department of Injury and Repair of Beijing Neurosurgical Institute from May to October 2004.Wistar rats were executed by dislocation and sterilized by putting into 75% alcohol. Four rats were used each time, the brain tissues of which were put into Hank's fluid. The cerebral pia mater and vessels were isolated under anatomical microscope, and the brain tissues were sheared off with eye scissors, which were filtered by 200 mesh copper grid and collected into 2 centrifuge tubes, and the supernatant was gotten rid off after that. ①Cells were divided into serum pre-culture group and control group according to whether there were serum pre-culture. The DMEM nutrient fluid of FCS (100 g/L) was added to serum pre-culture group, which was replaced by DMEM/F12 nutrient fluid of EGF, bFGF and B27 at 48 hours after culture. The cells in the control group were added with DMEM/F12 nutrient fluid of EGF, bFGF and B27 to culture in 5% CO2 incubator for one week at 37 ℃. The growth of NSCs at 48 hours after culture was observed in both groups under inverted contrast phase microscope.②On the 5th and 10th day after differentiation induced by 100 g/L FCS, nestin, GFAP, galactocerebroside and MAP2 were stained with immunofluorescence antibody,and the expressions of NSCs in both groups were observed under fluorescent microscope. The PBS buffer solution was used instead of first antibody in the control group, while other procedures were the same as above.MAIN OUTCOME MEASURES: ①Observation of NSCs growth at 48 hours after culture under contrast phase microscope in both groups. ②Detection of NSC specific antigen expressions with immunofluorescence stain in both groups.RESULTS: ①In the cells of the serum pre-culture group at 48 hours after culture, there were large number of regular NSC bulbs, most of which were aggregated by 8-10 cells. The proliferation of cell bulbs was accelerated after the nutrient fluid was replaced to DMEM/F12 nutrient fluid of EGF,bFGF and B27, while irregular lamellar cells could be seen in the control group at 48 hours after culture, and small regular cell bulbs were found at 4-5 days later. ②It could be seen under fluorescence microscope that on the 5th day after induced differentiation, the nestin, GFAP and galactocerebroside were positive, while MAP2 was negative. On the 10th day after induced differentiation, nestin and GFAP were positive, whereas the galactocerebroside and MAP2 were negative (no representation).CONCLUSION: Serum pre-culture can accelerate the bulb-aggregation of NSCs as well as promote the proliferation of NSCs.

In the present study carried out in our laboratory, we recorded local field potential (LFP) signals in primary visual cortex (V1 area) of rats during the anesthesia process in the electrophysiological experiments of invasive microelectrode array implant, and obtained time evolutions of complexity measure Lempel-ziv complexity (LZC) by nonlinear dynamic analysis method. Combined with judgment criterion of tail flick latency to thermal stimulus and heart rate, the visual stimulation experiments are carried out to verify the reliability of anesthetized states by complexity analysis. The experimental results demonstrated that the time varying complexity measures LZC of LFP signals of different channels were similar to each other in the anesthesia process. In the same anesthesia state, the difference of complexity measure LZC between neuronal responses before and after visual stimulation was not significant. However, the complexity LZC in different anesthesia depths had statistical significances. Furthermore, complexity threshold value represented the depth of anesthesia was determined using optimization method. The reliability and accuracy of monitoring the depth of anesthesia using complexity measure LZC of LFP were all high. It provided an effective method of realtime monitoring depth of anesthesia for craniotomy patients in clinical operation.
Anesthesia ; methods ; Animals ; Evoked Potentials, Visual ; Microelectrodes ; Monitoring, Physiologic ; Nonlinear Dynamics ; Photic Stimulation ; Rats ; Reproducibility of Results ; Visual Cortex ; drug effects

Information Systems ; Medical Informatics ; Medical Records Systems, Computerized ; Pathology

Objective To determine the optimum dose of pentazocine when combined with propofol for gastroscopy in elderly patients.Methods One hundred and forty ASA physical status Ⅰ or Ⅱ patients,aged 6575 yr,scheduled for elective gastroscopy under general anesthesia,were randomly assigned into Ⅰ-Ⅳ groups (n =35 each) using a random number table.Before insertion of the gastroscope,pentazocine 0.2,0.4 and 0.8 mg/kg were injected intravenously in Ⅱ-Ⅳ groups,respectively,while the equal volume of normal saline was given in Ⅰ group.Propofol was then administered by target-controlled infusion (TCI).The half-effective concentration (EC50 of propofol was determined by up-and-down sequential trial.The target plasma concentration (Cp) was set at 3.5 μg/ml in the first patient.Gastroscopy was performed at 5 min after the target effect-site and plasma concentrations were balanced.The response to gastroscopy was defined as positive when body movement and/or bucking occurred during gastroscopy.Each time the Cp increased/decreased by 0.3 μg/ml in the next patient depending on whether or not the response to gastroscopy was positive.EC50 and 95 ％ confidence interval of propofol TCI inhibiting the response to gastroscopy were calculated using Probit analysis.The development of respiratory depression and hypotension was observed.Results EC50 (95 ％ confidence interval) of propofol TCI inhibiting the response to gastroscopy was 2.82 (2.63-3.02) μg/ml,2.78 (2.58-2.97) μg/ml,2.16 (2.00-2.32) μg/ml and 2.03 (1.88-2.19) μg/ml in Ⅰ-Ⅳ groups,respectively.Compared with group Ⅰ,EC50 was significantly decreased in Ⅲ and Ⅳ groups,and the incidence of respiratory depression was increased in Ⅳ group (P ＜ 0.05).Conclusion The optimum dose of pentazocine when combined with propofol is 0.4 mg/kg for gastroscopy in elderly patients.

Objective To investigate the mechanism of cigarette smoke promotes non-small cell lung cancer (NSCLC) NCI-H520 cell line proliferation mediated by macrophages with method of blocking NFκB-p65 pathway by RNAi.Methods To co-culture NCI-H520 cells with primary macrophages or U937 cell line,the Transwell Inserts system was used in cell co-culture model.NFκB activation was confirmed by electrophoretic mobility shift assay (EMSA) and Western blot analysis.U937 cells were transfected with NFκB-p65 shRNA plasmid to abrogate the NFκB activation,by BrdU ELISA,the effect of cigarette smoke extract (CSE) promoted NCI-H520 cells proliferation were assessed,inflammatory factors TNF and IL-6 expressions were analysed by ELISA.Results Exposure of CSE enhanced NFκB-p65 nuclcus translocation and activated the NFκB pathway.CSE did not promote NCI-H520 cells proliferation alone (P ＞ 0.05),but after 4 days coincubation with macrophages,the proliferation of NCI-H520 cells was significantly increased (P ＜0.01),addition of CSE to the co-culture much more enhanced this effect (P ＜ 0.01).After NFκB-p65 was blocked by RNAi,it significantly reduced NFκB-p65 protein expression and inhibited NFκB activation in U937 p65-cells,and markedly inhibited U937 cells induced proliferation of NCI-H520 cells and IL-6,TNF secretion (P ＜ 0.01).Conclusion Cigarette smoke promotes NCI-H520 cells proliferation mediated by macrophages.Blockade of NFκB pathway with RNAi in macrophages can reduced cigarette smoke induced inflammatory factors secretion in macrophages,and significantly inhibit cigarette smoke promoted tumor proliferation.

BACKGROUND:Finding of neural stem cells(NSCs)brings new hope for repairing central nervous system(CNS)injury.However,the influence of internal environment after brain injury on the survival and differentiation of NSCs is a complexand variable process.OBJECTIVE:To observe the survival and differentiation of human embryonic NSCs following implantation into rats with fluid percussion brain injury.DESIGN:Open experiment.SETTING:Department of Neurosurgery,Guangdong Provincial Hospital of Traditional Chinese Medicine;Beijing Institute of Neurosurgery.MATERIALS:This experiment was carried out In the Laboratory of Neural Stem Cells,Beijing Institute of Neurosurgery from September 2002 to March 2003.Twenty-four female SD rats,aged 7 weeks,with body mass of(250±10)g,were provided by the Experimental Animal lnstitute, Chinese Academy of Medical Sciences[License Nd. SCXK(Jing)2002-2003]. Cerebrum of 8-week aborted fetus was obtained (Informed consents were obtained from parturients and their relatives). Fetal survival was monitored by B ultrasonic wave during abortion. BrdU monoclonal antibody(Sigma Company),rabbit anti-nidogen polyclonal antibody(Chemicon Company),mouse anti-microtubule-associated protein 2 (MAP-2)monoclonal antibody(Neomarkers Company),rabbit anti-gliaI fibrillary acidic protein(GFAP)polyclonal antibody (Biogenex Company).METHODS:① CerebraI cortex cells of 8-week aborted human fetus was harvested and cultured in vitro for obtaining human embryronic NSCs.②Rat models of hydraulic impact injury were developed.Bone window of motor sensory area of cerebral cortex was set at 2.5mm posterior to bregma which was zero point and 3.0 mm lateral to midline.Hydraulic impact injury parameters were set as impact pressure 0.3 MPa. impact time 25 ms and impact time once. ③BrdU-labeled human embryronic NSCs were implanted into injured area al 24 hours after injury.After 1 and 4 weeks.rats were sacrificed.Adjacent sections were doubly stained bv BrdU/MAP-2 and BrdU/GFAP.MAIN OUTCOME MEASURES:①Survival and immigration of implanted human embryonic NSCs.②Differentiation of implanted human embryonic NSCs.RESULTS:①BrdU-positive cells were oval and brown.At 1 and 4 weeks after implantation,BrdU-positive cells survived and migrated,and they migrated more widely at 4 weeks after implantation. ②At 1 week after implantation,more BrdU-positive cells were found in the subcortical granular layer and subcortex,and BrdU/MAP-2-positive cells were more than BrdU/GFAP-positive cells;At 4 weeks after implantation,BrdU-positive cells were significantly reduced,and found in choroid plexus and blood capillary,and BrdU/GFAP-positive cells were more than BrdU/MAP-2-positive cells.CONCLUSION:Implanted human embryonic NSCs can survive in the region of brain injury.gradually differentiate into astrocytes during rehabilitation and are easily digested by endothelial phagocytes.It indicates that immunological rejaction possibly influences the survival of human embryonic NSCs.

Objective To observe the influence of health education on compliance of children nn-dergoing megacolon radical operation. Methods A questionnaire investigation was conducted on compli-ance of children undergoing megncolon radical operation before and after health education. ResultsCompliance of children patients and their family members increased after health education (P<0.05).Conclusions Health education could improve the compliance of children patients and their family mem-bers as well as the cognition about the disease knowledge so that the rehabilitation was facilitated.