Objective:To investigate the role of mTOR in regulation of ICOS expression in human blood regulatory T cells. Methods:Isolation of Treg cells from human PBMC using MACS beads. We detected the ICOS expression on purified Treg cells and Treg cells viability using flow cytometry in anti-CD3 plus anti-CD28 ( antibody or beads) or anti-CD3 plus ICOSL-Fc for 3 days and 7 days. CFSE labeling human PBMC cells and in vitro cultured Treg mixed, Treg contact inhibition activity was detected by flow analysis. Results:After in vitro stimulation of Treg cells in the presence of anti-CD3+anti-CD28 for 3 days, there was no significant statistic difference in viability between ICOS+(92. 00±2. 69)% and ICOS-(90. 30±3. 53)% Treg-cells. After cultured for 7 days,the decreased ICOS+ Treg cells percentage within total Treg cells from ( 40. 20 ± 1. 83 )% to ( 11. 60 ± 1. 10 )% compared with that of 3 days. Further more,the ICOS expression level between stimulated with anti-CD28 or ICOSL-Fc condition group,compared with the ICOS MFI in the condition of anti-CD3 plus anti-CD28 treatment for 3 days was (2410. 0±746. 4) obviously higher than (403. 30±74. 42), that of the group treated with anti-CD3 plus ICOSL-FC. Rapamycin could partially suppress Treg cells ICOS expression,but unaffected the Treg suppression ability. Conclusion:ICOS expression level may not important for in vitro cultured human PBMC Treg cells survival although mTOR signling is important for regulation ICOS expression on in-vitro cultured Treg cells,but the ICOS expression on Treg regulated by multiply signaling pathways. CD28 signaling is the key stimulation factor for ICOS upregulation on in-vitro cultured Treg cells compared to ICOSL signaling.

Objective To study the inhibitory effects of pigment epithelium derived factor (PEDF) on oxygen-induced retinal neovascularization in mice,and to investigate the possible involvement of interleukin-1β (IL-1β) in the neovascular-inhibitory function of PEDF.Methods A total of 140 postnatal day (P)7 C57BL/6 mice were randomly divided into normal control group,oxygen-induced retinopathy (OIR) model group,PEDF treatment group and PBS treatment control group.All mice except normal control group with their mothers were exposed to (75 ± 2)％ oxygen environment for 5 days and then kept in room air for another 5 days to establish the OIR model.Mice in normal control group were kept in room air only.At P12 and P14,respectively,mice in PEDF treatment group received intravitreous injections of 1 μl PEDF (2 μg/μl),while PBS treatment control group received the same volume of PBS (10 mmol/L,pH7.4).All mice were euthanized at P17 and eyes were isolated.The changes of retinal vessels were observed on retinal flat mounts and cryosections by fluorescence microscopy.Retinal specimens were prepared for IL-1β protein and mRNA analysis by Western blot and real time fluorescence quantitative reverse transcription-polymerase chain reaction (Real-time RT-PCR).Results Changes of retinal vessels had been viewed by fluorescence microscopy on flat-mounted retina,the relative retinal neovascularization areas were significantly increased in OIR model group compared with normal control group (t =15.02,P＜0.01),and the relative retinal neovascularization areas were obviously smaller in PEDF treatment group than those in PBS treatment control group (t=5.96,P＜0.01).Fluorescence staining revealed that retinal vascular tufts were extending from outer plexiform layer (OPL) to ganglion cell layer (GCL) of the retina along with multiple interconnections; Neovascular tufts in OIR model group and PBS treatment control group were presenting distinctly more than those of normal control group and PEDF treatment group.The specific expression levels of IL-1β protein in retinas of OIR mice by Western blot analysis were higher than those of normal control group(t=3.35,P＜0.05),While these of PEDF treatment group showed a considerable decline in comparison with PBS treatment control group (P＜0.01),and there were no difference in normal control group and PEDF-treated group (F=11.764,P＞0.05).Similarly,expression levels of IL-1β mRNA tested by Real-time RT-PCR were obviously increased in the OIR model group when compared to normal control group(t =4.43,P ＜ 0.01).After treated with PEDF,expression levels of IL-1β mRNA showed a considerable decrease when compared to PBS treatment control group (P ＜ 0.01),and there were no difference in normal control group and PEDF-treated group (F=11.15,P＞0.05).Conclusions PEDF can inhibit oxygen-induced retinal neovascularization.The mechanism may be related to that PEDF can downregulate the expression of IL 1β in retina.

Objective To observe the in vitro damage to hairs by Trichophyton mentagrophytes and Microsporum canis using light microscopy and scanning electron microscopy ( SEM ), and to compare the differences in the duration needed for the two fungi to damage hairs in different age groups. Methods We collected healthy hairs from different age groups, and performed hair perforation test in vitro. The damage to the hairs was observed by SEM and light microscopy. Results Both T. mentagrophytes and M. canis could damage the hairs. The duration needed for T. mentagrophytes to damage the hairs was significantly shorter than that for M. canis in all age groups ( P

ObjectiveTo investigate the effects and indication of intermittent catheterization(IC) by relative safe bladder capacity(RSBC) to treat vesical-ureteral reflux(VUR).MethodsThere were 23 male complete SCI patients, aged between 7～45 years, including 15 patients without vesicao-ureteral reflux injuried 1～3 months as control group, 3 patients in mild group affected only ureter, and 5 patients in severe group affected not only ureter but also the pelvis. All the patients were taken intermittent catheterrization according to their own RSBC companied with supersonic device. The times of IC per day is that the average total urine volume one day was divided by RSBC. The observation time is over 2 years. During the oberservation some medicine was used to enlarge some patients bladder capacity.ResultsThere were no reflux in the control group; the reflux in the mild group improved a lot, some even disappeared, and there were no change in the severe group.ConclusionIC by RSBC is effective to prevent and cure vesical-ureteral reflux in some SCI patients.

Objective To investigate the adverse effects of nano-titanium dioxide (TiO2 )on kidney tissues in healthy rats and rats with oxidative stress (OS).Methods Thirty-six healthy male SD rats were randomly divided into control group, alloxan-treated group, nano-TiO2 treated (NM) group, and alloxan and nano-TiO2 dual treatment (OS-NM)group.Nano-TiO2 of three concentrations (0.5,5 and 50 mg/kg body weight)was injected intraperitoneally.The level of OS biochemical factors and blood urea nitrogen (BUN)and pathological changes of kidney were determined.Results Compared with those in NM and OS groups,the levels of O2 -· and superoxide dismutase (SOD)in OS-NM group were significantly increased after exposure to nano-TiO2 of 5 mg/kg body weight (P <0.05).Nano-TiO2 of 50 mg/kg body weight led to significant changes of O2 -·,SOD,and glutathione (GSH) levels in OS-NM group in comparison with OS and NM groups (P <0.01).The levels of O2 -· and GSH between OS group and NM group changed significantly (P < 0.05 ).Compared with healthy rats,OS rats showed significant increased BUN level (P <0.01),cell number and edema of renal tubular epithelial cells after nano-TiO2 exposure.A synergic effect between OS condition and nano-TiO2 level was shown on the increased level of O2 -·.Conclusion Nano-TiO2 induced more adverse effects on the kidney in OS rats,suggesting a synergistic effect between nano-TiO2 and OS.This result provides experimental evidence for patients’safe use of nano-TiO2 .

Objectives To summarize and analyze the characteristics ofspina bifida occulta in prenatal ultrasound scan in order to improve the accuracy of diagnosis.Methods Sixteen singleton pregnant women with fetal spina bifida occulta who were diagnosed in the Ultrasonography Department of Dalian Maternal and Child Health Hospital from January 2014 to April 2015,were enrolled in this study.Diagnosis was made based upon the abnormalities found in the spinal column of fetus during routine ultrasound scan,followed by a close examination on the fetal spinal column and the position of conus medullaris by using multiple scan.Descriptive analysis was used to summarize the characteristics of spina bifida occulta in ultrasonic images.Results Among the 16 cases,12 had subcutaneous mass (nine cases had follicle-like anechoic mass and three cases had masses with strong echoes).The other four cases without subcutaneous mass manifested signs of disappeared physiological curvatures or disordered arrangement of spinal column,angulation deformity or scoliosis,or asymmetry in ossification centers of spinal column on both sides of lesions and lowered position of conus medullaris.Lemon shaped head and banana shaped cerebellum were seen in one case,while the other 15 cases were found no cerebral abnormalities.Eight cases had tethered cord (the position of conus medullaris was lower than the second lumber vertebra),six cases had unclear position of conus medullaris and the other two were normal.Diagnoses for all of the 16 cases were confirmed by MRI after delivery or autopsy or neonatal operation.Conclusions Abnormalities such as subcutaneous mass and tethered cord observed in prenatal ultrasound scan are significant indicators for suspected closed spinal dysraphism and further multiple scan is required to confirm the diagnosis.

OBJECTIVE To study and find reasonable methods of nursing care of incision through analyzing the risk factors in liver transplant patients.METHODS A total of 416 liver transplantation operations in our hospital from Sep 2004 to May 2006 were evaluated.The risk factors that resulted in the incision infection were considered.RESULTS The incision infection rate was decreased from 13.9% to 4.3% after improvement.CONCLUSIONS Nursing care is of great significance in preventing the incision infection in liver transplant patients.

Objective: To study the related factors of radiation induced lung injury(RILI) after three-dimensional conformal radiotherapy(3D-CRT) in the elderly patients with advance esophageal cancer, thus to provide reference for radiotherapy planning. Methods: The clinical data and physical parameters of 72 elderly patients with advance esophageal cancer were analyzed, including gender, age, performance status scoring, smoking history, tumor location, T staging, underlying disease of the lung, radiation dose, the whole lung accepted 5 Gy exposure volume(V5), V10, V15, V20, V25, V30, V40 and mean lung dose(MLD). Results: Among 72 patients, 15 cases developed RILI.It was revealed by univariate analysis that there were statistically significant differences in advanced age, underlying disease of the lung, V5, V10, V15, V20, V25, MLD with RILI(χ²=5.098, P=0.026; χ²=3.598, P=0.030; t=3.854, P=0.034; t=4.901, P=0.022, t=4.638, P=0.029, t=5.122, P=0.015, t=3.652, P=0.041, t=5.760, P=0.010). Conclusion It should be payed more attention to factors such as advanced age, underlying disease of the lung, V5, V10, V15, V20, V25 and MLD when elderly patients with advance esophageal cancer patients were treated with 3D-CRT in order to prevent and decrease the risks of RILI.

Objective To generate of human induced pluripotent stem cells from umbilical cord matrix cells(UMC).Methods Sox2 and Klf4 and Oct4 and c-Myc were transfected into UMC cells with retrovirus,and thcn UMC cells was reprogrammed to iPS cells.Gene expression was confirmed with RT -PCR and the integration was confirmed with cell karyotype.iPS cells were further validatcd with cell alkaline phosphatase detection and immunofluorescence staining,differentiating into teratomas in vivo and embryoid bodies in vitro.Results iPS cells were similar to embryonic stem cells (ES).The expression of Nanog,Oct4,Rex1 and Sox2 in iPS cells were higher than UMC cells,and Sox2,Klf4,Oct4,c-Myc silenced in iPS cells.Exogenous genes were inserted into the nucleus of iPS cells,which was confirmed by 1％ agarose gel electrophoresis.iPS ccll karyotype was normal,alkaline phosphatase activity increased,ES cell-specific proteins,including SSEA3,SSEA4,TRA-1-60,TRA-1-81 and Nanog,were expressed.iPS cells were differentiated into a teratoma in vivo and embryoid bodies in vitro.Conclusions iPS cells were similar to ES cells,which had pluripotency.

Objective To investigate the risk factors and outcomes of acute kidney injury (AKI) in patients after intra--coronary stent implantation.Methods A retrospective and case control study was done with data analysis in 325 patients who underwent intra-coronary stent implantation from January 2010 to March 2011.The patients were divided into two groups as per the criteria of AKI identified on the 7th day after implantation of stent.The variables to be studied included:(1) age,gender,hypertension,diabetes,cerebrovascular disease,left ventricular insufficiency,peripheral angiopathy,creatinine,urea nitrogen,estimated glomerular filtration rate,hyperuricemia,proteinuria,emergency operation,hydration,and medication (ACEI/ARB,statins) before operation; (2) dose of contrast media,operation time,hypotension during intra-operative period; and (3) postoperative:hypotension.The variables were analyzed with the process of One-way ANOVA and multivariate Logistical regression analysis.Consequently,the independent risk factors of AKI in patients after intra-coronary stent implantation could be found.Further,the prognosis of AKI patients was analyzed.Results Of the 325 patients,51 (15.7％) developed AKI.Compared the normal group,hospital stay (P ＜ 0.01 ) and in-hospital mortality (P ＜ 0.05) increased significantly in the AKI group.Monofactorial analysis showed that age,pre-operative laboratory and clinical data including left ventricular insufficiency,peripheral angiopathy,creatinine,urea nitrogen,estimated glomerular filtration rate, hyperuricemia, proteinuria, hydration and emergency operation, and intraoperative information such as operation time and hypotension,and postoperative hypotension in AKI patients group were significantly different in comparison with control group ( P ＜ 0.05 ). Multivariate logistic regression analysis revealed that elderly age (OR =0.253),pre-operative proteinuria (OR =5.351 ),preoperative left ventricular insufficiency ( OR =8.704),eGFR ≤ 60 ml/ ( min · 1.73 m2 ) ( OR =6.677 ),prolonged operation time ( OR =1.017),intra-operative hypotension ( OR =25.245 ) were independent risk factors of AKI ( P ＜ 0.05 ).Conclusions AKI is a common complication and associated with increase in mortality after intra-coronary stent implantation.Increase in age,pre-operative proteinuria,pre-operative left ventricular insufficiency,pre-operative low estimated glomerular filtration rate,prolonged operation time,intra-operative hypotension are the independently risk factors associated with AKI.