Objective To evaluate the safety and effect of laryngeal mask airway (LMA) general anesthesia in neonatal laparoscopic pyloromyotomy. Methods The clinical data of 100 neonates who had underwent laparoscopic pyloromyotomy were retrospectively analyzed. The neonates were divided into LMA group (50 cases) and tracheal intubation group (50 cases) according to the anesthesia method. The heart rate and mean arterial pressure (MAP) were recorded at the time points of preanesthesia (T0), tracheal intubation (T1), 10 min after tracheal intubation (T2) and extubation (T3), and the extubation time, and consumption of sevoflurane and postoperative complications were also recorded. Results All the 2 groups had smooth intubation and LMA, and the operation was completed successfully under laparoscope. There was no conversion to laparotomy. In LMA group, there were no statistical difference in heart rate and MAP at all the time points (P>0.05). The heart rate and MAP at T1 and T3 in tracheal intubation group were significantly higher than those at T0, and there were statistical differences (P<0.05). The heart rate and MAP at T1 and T3 were significantly lower than those in tracheal intubation group, heart rate:(131.1 ± 11.0) times/min vs. (146.9 ± 13.7) times/min and (131.9 ± 9.8) times/min vs. (147.3 ± 14.6) times/min; MAP: (44.2 ± 5.5) mmHg (1 mmHg = 0.133 kPa) vs. (47.9 ± 8.4) mmHg and (45.1 ± 7.3) mmHg vs. (49.1 ± 9.4) mmHg, and there were statistical differences (P<0.01 or<0.05). The extubation time, consumption of sevoflurane and incidences of postoperative complications in LMA group were significantly lower than those in tracheal intubation group:(6.1 ± 2.2) min vs. (12.5 ± 3.6) min, (2.9 ± 1.1) ml vs. (5.1 ± 1.9) ml and 4％ (2/50) vs. 48％ (24/50), and there were statistical differences (P<0.01). Conclusions LMA anesthesia in neonatal laparoscopic surgery is activated to maintain the steadiness of hemodynamics, which has less stress, anesthetic drugs consumption and respiratory complications compared with tracheal intubation .

Ras/Raf/MEK/ERK singal transduction plays an important role in cell proliferation, differentiation, apoptosis, metastasis and metabolism. This investigation focused on this signal pathway and chose farnesyl transferase (FTase) as the main target and Raf-1 kinase as the second target. A lot of compounds were selected to construct the pharmacophore models of farnesyl transferase inhibitors (FTIs) and Raf-1 kinase inhibitors by using computer-aided drug design (CADD). The pharmacophore of FTIs is constituted by a hydrogen bonding acceptor, an aromatic ring, a positive ionizable and two hydrophobic regions; the pharmacophore of Raf-1 kinase is constituted by a hydrogen donor, a hydrogen acceptor, a hydrophobic regions and an aromatic ring. There are some similarities between the two pharmacophores. After analysis of the constructions of these two pharmacophores, some new aminomethylbenzoic acid derivatives with good forecasting activity against both of FTase and Raf-1 kinase were designed with these new pharmacophore models.

Objective: To use vertical and humidity correcting method to calibrate aerosol optical depth (AOD), and to explore the feasibility of calibrated-AOD as exposure index to measure the level of air pollutants from the ground and to establish the exposure-response relationship between calibrated-AOD and people's health outcomes.Methods: First of all, we interpolated AOD, PM2.5, relative humidity and planetary boundary layer height using Kriging method to obtain data at different locations and matched different data with respiratory death in Beijing by geographical coordinates.Then, the planetary boundary layer height and aerosol hygroscopic growth factor calculated based on relative humidity was used to calbrate the AOD.To compare the effects of calibrated-AOD and PM2.5, we used standardization method to get non dimensionless calibrated-AOD and PM2.5.At last, we used the generalized additive mixed model (GAMM) to estimate the acute effects of calibrated-AOD and PM2.5 on respiratory death and chronic obstructive pulmonary disease (COPD) death, after controlling the time trend, temperature and humidity effects, days of the week effect and holiday effects.Results: The correlation coefficient between calibrated-AOD and PM2.5 was 0.72.The effects of calibrated-AOD on respiratory death and COPD death was the strongest at lag 0-3, one unit of calibrated-AOD increases was associated with 3.64% (95%CI: 0.58%-6.78%) increase of respiratory death and 4.92% (95%CI: 1.81%-8.14%) increase of COPD death.As for PM2.5, the strongest effects appeared at lag 0-1, one unit of PM2.5 (about 155 μg/m3) increases was associated with excess risks of 3.96% (95%CI: 0.82%-7.19%) and 6.12% (95%CI: 1.44%-11.02%) for respiratory death and COPD death respectively.Compared with PM2.5, the effects of calibrated-AOD on respiratory death and COPD death had narrower confidence intervals.The calibrated-AOD was sensitive to capture the lag effects, and the cumulative lag effects of calibrated-AOD were all significant on multiple lag days which indicated that the calibrated-AOD was sensitive to capture cumulative lag effects of air pollutants on respiratory death and COPD death as well.Conclusion: We believe that calibrated-AOD can be used as an index to reflect the effects of air pollutants on respiratory death in Beijing.In the absence of ground monitoring, calibrated-AOD can be used to mea-sure the relationship between air pollutants and some health outcomes.

Objective To clone platelet-derived growth factor A chain (PDGF-A) gene and insert PDGF-A gene into. Enhanced green fluorescent protein (EGFP) vector and then transformed into dermis-drived mesenchymal stem cells (DMSCs). Methods cDNA clones encoding human PDGF-A gene were isolated from a human hepatoma cell line mRNA by reverse transcription-polymerase chain reaction (RT-PCR). The PCR amplified fragment of PDGF-A gene was cloned into pMD18-T vector. The eukaryotic expression vector pEGFP-N1/PDGF-A was constructed by subcolone PDGF-A gene into pEGFP-N1 vector. PDGF-A gene was transfected into DMSCs with the help of Fugene 6 transfection reagent. Results Full cDNA sequence encoding human PDGF-A gene had been cloned, which sequence was consistent with the reported sequence in GenBank by sequence assaying. Conclusion cDNA sequence encoding human PDGF-A gene was successfully cloned into pEGFP-N1. The transient expression of PDGF-A gene in DMSCs has been realized.

Objective To explore the protective effect of atorvastatin on irradiated endothelium and the thrombomodulin(TM)expression.Methods Cultured human coronary artery endothelial cells(HCAEC)and human umbilical vein endothelial cells(HUVEC)were treated by atorvastatin at the final concentration of 10 μmol/ml for 10 min,and then irradiated with 2 and 25 Gy.Cell cycles status and TM expression were quantitatively measured by flow cytometry 24 hours after irradiation.Protein C activation in endothelial cells was also assessod.Results After administration with atorvastatin for 24 h,the TM expression increased by 77%,59% and 61% in normal control group,2 Gy group and 25 Gy group,respectively(t=27.395,26.420,58.065;P=0.000).The protein C levels decreased by 23% and 34% compared with the normal group post-irradiation to 2 and 25 Gy,but increased by 79% and 76% compared with the irradiated control group after administration with atorvastatin.The rates of cell apoptosis decreased by 6% and 16% in 2 Gy and 25 Gy groups,respectively after administration with atorvastatin for 24 h(t=4.178,17.863;P=0.000).Conclusions Atorva statin can protect endothelia cell from irradiation-induced apeptosis by increasing TM expression and protein C activation.

Multi-target drugs attract increasing attentions for the therapy of complicated neurodegenerative diseases. In this study, a computer-assisted strategy was applied to search for multi-target compounds by the pharmacophore matching. This strategy has been successfully used to design dual-target inhibitor models against both the acetylcholinesterase (AChE) and poly (ADP-ribose) polymerase-1 (PARP-1). Based on two pharmacophore models matching and physicochemical properties filtering, one hit was identified which could inhibit AChE with IC50 value of (0.337 +/- 0.052) micromol x L(-1) and PARP-1 by 24.6% at 1 micromol x L(-1).

Objective To explore the roles of PKCθ(protein kinase Cθ)signaling pathway on the activation,proliferation and TH1/TH2 cytokines production of the γδT lymphocytes stimulated by Mycobacterium tuberculosis antigen(Mtb-Ag) in vitro.Methods Peripheral blood mononuclear cells(PBMC)were pretreated with or without Rottlerin at 5.0 μmol/L,and stimulated with Mtb-Ag and cultured in rhIL-2 containing medium.After different time of culture,activation molecules and cytokines production of γδT cells were measured by flow cytometry.The proliferation proportion and the percentage of each generation of γδT cells were determined by carboxylfluoreseein diacetate succinmidyl ester(CFSE)labeling technique and flow cytometry.Results After 3d of stimulation with Mtb-Ag,the expression rates of CD69 and CD25 of γδT cells were 46.2%and 45.6%,respectively.Pretreatment of 5.0 μmol/L Rottlerin markedly inhibited the both expressions of CD69 and CD25 in γδT cells(P＜0.01).After stimulation and expansion in vitro for 5,10,and 15 d,the percentages of the γδT cell were 9.6%,54.6%and 82.4%,respectively.There was a few γδT cells in propagation on the 5th day of culture,and almost all γδT cell divisions were above 6 generations on the 10th and 15th day of culture.Pretreatment of the Rotflerin significantly suppressed the γδT cell proliferation,but after 10 d culture,there were still a few parts of γδT cells in propagation.Meanwhile,after 7,14,and 21d of culture,upon stimulation with PMA+Ionomycin,the IFN-γ producing-γδT cells were about 80%at all times.But only after 21d culture,IL-4-producing γδT cells was 2.6%.,The percentage of IFN-γ producing γδT cells markedly reduced in Rottlerin group(P＜0.01).IL-4 secretion of the γδT cells was almost completely blocked.Conclusion PKCO signal pathway involves in activation,proliferation and differentiation of the γδT lymphocytes stimulated by Mtb-Ag.

Objective To explore the value of 1H-MRS on the evaluation of Alzheimer's disease (AD) with neural stem cells (NSCs) transplantation in an APP-PS1 double transgenic (tg) AD mouse model.Methods NSCs from C57BL/6 mice were cultured and amplified.APP-PS1 tg mice (n =30) aged 12 months were used as the study group,and mild-type mice (n =15) were used as the control group.Animals in the study group were randomized into two subgroups,the AD mice in one subgroup received NSCs transplantation (NSCs group) and in another subgroup received phosphate buffer saline (PBS,PBS group)in bilateral hippocampal CA1.Animals in the control group were not treated.Using a 7.0 T high-fieldstrength MR imager,1H-MRS was performed before and 6 weeks after transplantation to measure the area under the peak of n-acetyl aspartate (NAA),glutamate (Glu),myo-inositol ( mI),choline (Cho) and creatine (Cr) in the hippocampal area,NAA/Cr,Glu/Cr,mI/Cr and Cho/Cr ratio were calculated and compared with histopathological results (including Nissl's staining and electron microscope examination).Comparisons among NSCs,PBS and control groups were conducted by one-way ANOVA.Results NSCs from C57BL/6 mice were cultured successfully. Before transplantation,the mean NAA/Cr,Glu/Cr and mI/Cr in NSCs,PBS and control groups were 0.89 ± 0.05,0.88 ± 0.04 and 1.15 ± 0.05,0.40 ± 0.03,0.39 ± 0.03 and 0.45 ± 0.05,0.67 ± 0.05,0.67 ± 0.05 and 0.52 ± 0.04,respectively,and differences were statistically significant (F =148.918,7.529,59.468,P ＜ 0.01 ). There were no significant differences in NAA/Cr,mI/Cr and Glu/Cr ratios between NSCs and PBS groups before transplantation (t =0.147,0.096,0.207,P ＞ 0.05 ),but the differences were significant compared with the control group (t =0.255,0.467,0.171 and t =0.269,0.527,0.151,P ＜0.05).Six weeks after transplantation,the mean NAA/Cr,Glu/Cr and mI/Cr in three groups were 1.13 ±0.07,0.86 ±0.05 and 1.14 ±0.05,0.45 ± 0.04,0.38 ± 0.02 and 0.44 ± 0.03,0.58 ± 0.04,0.67 ± 0.04 and 0.53 ± 0.04,respectively,and differences were statistically significant ( F =112.092,23.076,44.367,P ＜ 0.01 ).NAA/Cr and Glu/Cr ratios were increased and mI/Cr was decreased in NSCs group,and the difference was significant compared with PBS group at the same time point ( t =0.271,0.071,0.089,P ＜ 0.05 ).There were no significant differences in NAA/Cr and Glu/Cr ( t =0.013,0.012,P ＞ 0.05 ),but there was a significant difference in mI/Cr between NSCs and control groups ( t =0.046,P ＜ 0.05).There were no significant differences in Cho before and after transplantation among the three groups (P ＞ 0.05 ). Nissl's staining showed that the number of neurons in the hippocampal area increased more significantly in tg mice receiving NSCs than that without receiving NSCs.Electron microscopy showed that most hippocampal NSCs in NSCs group were morphologically normal with abundant organelles,while hippocampal NSCs in PBS group were swollen with sparse synapses.Conclusion 1H-MRS is able to display intracranial metabolite changes before and after NSCs in APP-PS1 double transgenic AD mice and has an applicable value in evaluating the therapeutic effect of NSCs on AD.

Objective To explore changes of metabolites in APP/ PS1 double transgenie mice of Alzbeimer disease (AD) by 1H-MR spectroscopy (1H-MRS) and the application value of in early diagnosis of AD.Methods 1H-MRS was performed in 35 APP/PS1 transgenie mice of AD ( study group) and 20 wild type mice ( control group) at age of 3, 6 and 9 months using a 7.0 T MR system.Sub-peak areas of N-acetyl aspartate (NAA), myo-inositol (mI) and creatine (Cr) in the cerebral cortex and hippocampus were measured, and the NAA/Cr and mI/Cr ratios were calculated.The changes in pathology between the two groups were compared.Using the lower limit of 95% confidence interval (CI) of the ml/Cr ratio and the upper limit of 95% CI of the NAA/Cr ratio of AD mice as the threshold, their influences on sensitivity,specificity and accuracy of various age groups of AD animals were compared.Comparison of the 1H-MRS indexes between study mice and wild type mice at each time point were conducted by a two-sample t test.Results The mean mI/Cr ratios of AD mice were 0.68± 0.03, 0.72± 0.04, and 0.77 ± 0.04 respectively at 3, 6 and 9 months of age; while they were 0.63 ± 0.04, 0.64 ± 0.03, and 0.64 ± 0.04 respoetively in control group, the difference was significant ( t = 2.814, 5.146, 14.437, P < 0.01 ).Compared with the control group, the mI/Cr ratio of the 3-month-old AD mice of the study group was significantly increased,and histological examination showed proliferation and activation of neuroglial cells in the cerebral cortex and hippoeampus.The mean NAA/Cr ratio were 1.17 ±0.08, 1.04 ±0.05, and 0.90 ±0.05 respectively at 3,6 and 9 months of age in study group, while they were 1.18 ±0.07, 1.16 ±0.07, and 1.18 ±0.08respectively in control group.There were no significant difference ( t = 0.752, P > 0.05 ) between the study group and control group at 3 months of age, and the NAA/Cr ratio decreased significantly only at 6 and 9 months of age ( t = - 8.514, - 5.646, P < 0.01 ).The immunohistochemical exam demonstrated the appearance of Aβ plaque.According to threshold of mI/Cr, the sensitivity of AD mice of 3, 6 and 9 months of age was 80% (28/35), 84% ( 26/31 ) and 85% ( 23/27 ), and the specificity was 85% ( 17/20 ),94% (17/18) and 100% ( 16/16), and the accuracy was 82% (45/55), 88% (43/49) and 91% (39/43),respectively.For NAA/Cr, the sensitivity of AD mice of 6 and 9 months of age was 84% (26/31) and 89% (24/27), and the specificity was 89% (16/18) and 100% (16/16), and the accuracy was 86% (42/49) and 93% (40/43), respectively.Conclusions NAA and mI are the most sensitive and specific markers for early assessment of AD, and change of mI is earlier than that of NAA.Quantitative analysis of mI may provide important clues for early diagnosis of AD.

Objective To label neural stem cells (NSCs) with superparamagnetic iron oxides (SPIO) and to explore the tropism of NSCs after transplantation into the hippocampus of APP/PS1 AD mice by MRI.Methods NSCs from C57BL/6 mouse were cultured and identified.Feridex and Poly-L-Lysine were added into the medium to be co-cultured to make magnetic labeled NSCs and transmission electron microscopy was used to identify the iron particles in NSCs.Transgenic (tg) and wild-type (wt) mice at 12 months of age were divided into three groups: SPIOs labeled NSCs group (A and C),unlabeled NSCs group(B).Feridex-labeled NSCs were migrated into the hippocampus of APP/PS1 AD mice to monitor in vivo by MRI.After 1,2,4 and 6 weeks,the mice were sacrificed and their brain tissues were sectioned to investigate the migration of SPIO labeled NSCs and compared with MRI.Results NSCs of C57BL/6 mice were cultured successfully.Transmission electron microscope showed visible iron granules in cytoplasm.MRI detection of labeled cells: T2WI and T2* WI showed remarkable low signal intensity at the hippocampus injection points 1 week after transplantation,particularly on T2* WI.Area of low signal intensity enlarged increasingly along the injection points after 2 weeks.At 4 weeks,area of low signal intensity spread throughout the hippocampus,but intensity shadowed Six weeks later,low signal intensity almost disappeared.There was no obvious low signal change in unlabeled cell transplantation group.For wt mice,size and location of low signal did not appear obvious change at all designated time points.Prussian blue positive cells were observed in the hippocampus,indicating that NSCs labeled with SPIO could survive,migrate and differentiate in the brain of the APP/PS1 AD mice.Changes of pathology were well correlated with the area where a signal intensity loss was observed in MRI 1,2,4 and 6 weeks after transplantation Conclusions Diffuse migration of transplanted NSCs labeled with SPIO is observed in the hippocampus in APP/PS1 tg mice,and MRI technique is an ideal method for tracking labeled stem cells after grafting in vivo.