Objective To observe the influence of Fibronectin-Thrombopoietin(FN-TPO) gene modified human bone marrow mesenchymal stem cells(MSCs) on the engraftment of cord blood hematopoietic stem cells.Methods FN-TPO gene modified human bone marrow MSCs combined with cord blood mononuclear cells(CB-MNC) were transplanted to sublethal dose treated severe combined immunodeficiency disease(SCID) mice.After transplantation,these mice were observed for 4 weeks.Peripheral blood cell counts were performed at different time point to assay the hematopoietic system status of the mice.Four weeks after the transplantation,human-sourced cell integration was assayed by flow cytometry(FCM) and polymerase chain reaction(PCR).Results One week after the cell transplantation,every main index of the peripheral blood cell counts in the gene modified group was higher than that in the control groups(P

Background Cardiomyocytes apoptosis was related to oxidative stress which has been shown to play an important role in ventricular remodeling after myocardial infarction(MI).Probucol is a hypolipidemic and antioxidant agents.Several reports demonstrated its cardioprotective effect after MI.However,the exact effect of probucol on the modification of the apoptosis related gene Bcl-2 and Bax is not clear.Objective To investigate the effects of probucol on mRNA and protein expression of Bcl-2,Bax and oxidative stress in myocardial infarcted rats.Methods Forty-one SD rats that survived 24 h after ligating left anterior descending coronary artery were randomly to receive placebo-saline(5 mL/d,n=20)or probucol(probucol 60 mg/kg?d,n=21).Twelve rats underwent sham operation were served as control(n=12).Six weeks after treatment,hemodynamic pararmeters and left ventricular function were measured with catheterization.Cardiomyocytes apoptosis were determined by TUNEL method.Myocardium mRNA of Bcl-2 and Bax expression levels in the non-infarcted myocardium were as- sessed by RT-PCR.The myocardium protein expression of Bcl-2 and Bax in the non-infarcted myocardium were de- termined by Western blot.Colorimetry was used to determine oxidative metabolism index in myocardium.Results 1)Compared with the sham rats,all MI rats showed marked decrease in Bcl-2 mRNA and protein expression in myo- cardium with increase of Bax mRNA and protein expression and apoptosis index(P

Objective To detect the expressions of miR-155,T helper type 17 (Thl7) cells,and Th17 cellspecific transcription factor RORγt and effector cytokine interleukin (IL)-17 in peripheral blood and skin lesions from,and to evaluate their relationship in,patients with atopic dermatitis (AD).Methods Peripheral blood was obtained from 37 patients with AD and 33 age-and sex-matched healthy controls,and biopsy specimens from the lesional and perilesional skin of five patients with severe AD as well as from the normal skin of five healthy human controls.Real-time fluorescence-based reverse transcription (RT)-PCR was employed to measure the mRNA expression levels of miR-155,RORγt and IL-17 in peripheral blood mononuclear cells (PBMCs) and skin specimens,flow cytometry to detect the percentage of Th17 cells in PBMCs,enzyme-linked immunosorbent assay (ELISA) to determine the plasma concentration of IL-17.Statistical analysis was done using independent sample's t test,one-way analysis of variance followed by the least significant difference test,and linear correlation analysis.Results Compared with the healthy controls,the patients with AD showed a significant increase in Th17 cell percentage (1.78％ ± 0.52％ vs.0.47％ ± 0.15％,P＜ 0.01),mRNA expression levels of miR-155 (5.78 ± 1.78 vs.1.82 ± 0.46,P＜ 0.01),RORγt (6.08 ± 1.04 vs.1.64 ± 0.52,P＜ 0.01) and IL-17 (7.09 ± 1.75 vs.1.71 ± 0.46,P＜ 0.01),as well as in the plasma concentration of IL-17 ((2.51 ± 6.15) pg/ml vs.(11.80 ± 2.24) pg/ml,P＜ 0.01).There was a sequential decrease in the expression levels of miR-155,RORγt and IL-17 mRNA from lesional skin,perilesional skin to normal skin (F =41.803,17.040 and 37.064 respectively,all P ＜ 0.01).The miR-155 mRNA expression level in PBMCs was positively correlated with the SCORing Atopic Dermatitis (SCORAD) index,Th17 cell percentage,RORγt and IL-17 mRNA expression levels as well as IL-17 plasma concentration (r =0.405,0.426,0.402,0.410 and 0.408 respectively,all P ＜ 0.05).Similarly,the miR-155 expression level was positively correlated with RORγt and IL-17 mRNA expression levels in lesional and paralesional specimens (r =0.428 and 0.435 respectively,both P ＜ 0.05).Conclusion The up-regulated expression of miR-155,Th17 cells and their effector cytokine IL-17 may be associated with the development of AD.

AIM: To make a contrast and then analyze the difference of optical coherence tomography (OCT) before and after vitreomacular traction syndrome (VTS) was performed.METHODS: The clinical date of 11 evaluable eyes of 11 patients with VTS who were diagnosed by OCT and underwent 25G vitreous surgery from January 2013 to January 2014 were retrospectively analyzed.Patients were followed up for an average of 6mo, to observe the visual acuity and OCT examination of the patient before and after operation.We compared the changes of retinal thickness and local morphology before and after operation.RESULTS: After vitreous retraction, 6 eyes improved, 2 eyes do not improve.One eye received macular membrane traction, in the operation the macular epiretinal membrane peeling, retrial membrane stripping and the triamcinolone acetonide intravitreal injection were given, but after the operation, the vision does not improve.Two eyes received vitreous combined with retinal macular membrane traction.In the operation, macular epiretinal membrane stripping was given, after the operation, visual acuity improved.The proportion of those with visual acuity of 0.1 or more increased from 46% before to 73% after the operation.Before operations, the mean central macular thickness was 619.27±195.13μm, 239.12±143.84μm after, which decreased significantly (P<0.05).CONCLUSION: Vitrectomy can effectively relieve the vitreous traction of the macula, and can prevent further decline in visual acuity and reduce macular edema as well as improve the visual acuity of some patients.So, OCT has important guiding significance on the diagnosis and prognosis of this group.

Objective To observe the lung protection of Astragalus membranaceus against radiotherapy to intermediate-stage and terminal thoracic neoplasm, and its influence on TNF-α and ET expression.Methods The patients with intermediate-stage and terminal thoracic neoplasm under radiotherapy were divided into a treatment group and a control group.Patients in the treatment group took 10 ml of Asragalus membranaceus twice a day.for consecutive 6 months from the beginning of radio therapy.TNF-α and ET in the plasma were measured before and after the radiotherapy.The clinical symptom,iconographic changes and lung diffusion were observed from the 15th day of radiotherapy.Results The TNF-α and ET in plasma afterthe radiotherapy were(2.48±0.75)as/ml and(69.32±23.03)pg/ml for the treatment group,and(5.12±1.01)ns/ml and(97.87±37.83)pg/ml for the control group with the statistial difference(x2=7.49,6.57,P<0.001).The decrease of CO diffusion 5 and 10 months after the radiotherapy in the treatment group was statistically different compared with that in the control group(x2=3.98,3.78,P<0.05).There was a statistical difference of the incidence of acute radiation pneumonitis and pulmonary fibrosis between these two groups(P<0.05).Conclusions Astragalus membranaceus could inhibit the excess expression of TNF-α and ET in plasma and reduce the deterioration of diffusion after radiotherapy,so that it can be used for intervention of lung injuries from radiotherapy.

Objective To observe the EGFR nuclear translocation in cervical carcinoma cell lines after irradiation and its possible role in radiation tolerance.Methods Western blotting was used to detect the nuclear EGFR and cytoplastic EGFR after irradiation.The effect of Cetuximab on expression of nuclear EGFR and survival fractions were investigated.Results After irradiation,compared with control group,the expression of nuclear EGFR protein increased in irradiated cervical carcinoma cell.Cetuximab inhibited the radiation-induced nuclear EGFR expression with decreased survival fractions.Conclusion Radiation could induce EGFR nuclear translocation in cervical carcinoma cell lines and nuclear EGFR might be correlated with radiation tolerance in Cervical carcinoma cell.

Objective:To determine the effect of Notch1 signaling pathway on the differentiation and function of Th17 cells in murine psoriasis model.Methods: BALB/c mice were randomly divided into psoriasis model group and control group.Murine psoriasis model was established by topical 5% imiquimod application in combination with intraperitoneal injection of α-2b interferon.The CD4+ T lymphocytes were isolated by magnetic activated cell sorter (MACS).Flow cytometric analysis (FCM) was performed to detect the percentage of Th17 cells.Real-time RT-PCR was employed to measure the mRNA levels of RORγt,IL-17A,Notch1 and Hes-1.The CD4+ T lymphocytes were then divided into γ-secretase inhibitor DAPT groups and control group,and the expression differences of Notch1 signaling molecule and its target gene Hes-1 mRNA levels,Th17 cell percentage,RORγt and IL-17A mRNA levels,and IL-17A concentrations in cell-free supernatant were detected.Results: The expression levels of Th17 cell percentage and RORγt,IL-17A,Notch1 and Hes-1 mRNA in CD4+ T lymphocytes of murine psoriasis model were significantly higher than control mouse[(2.97±0.86)% vs.(0.65±0.11)%,t=15.083;(5.75±0.61) vs.(1.57±0.43),t=21.630;(7.83±0.97) vs.(1.63±0.31),t=25.348;(7.10±1.37) vs.(1.47±0.34),t=17.386;(7.30±1.15) vs.(1.67±0.48),t=18.840,respectively,all P<0.01].Compared with control group,Th17 cell percentage,mRNA expression levels of Notch1,Hes-1,RORγt and IL-17A,and IL-17A concentrations in cell-free supernatant from cultured CD4+ T lymphocytes of murine psoriasis model were dramatically decreased in DAPT treated groups in a dose-dependent way (F=74.368,89.719,126.572,94.558,124.323 and 123.231 respectively,all P<0.01).Conclusion: Notch1 signaling pathway can regulate the differentiation and function of Th17 cells in murine psoriasis model,and may have potential value for the target immunotherapy of psoriasis.

Animals ; Animals, Newborn ; Cells, Cultured ; Fibroblasts ; cytology ; metabolism ; Myocardium ; cytology ; Pravastatin ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1 ; genetics ; metabolism

Objective To establish an simple and convenient animal model of cardiac arrest for studying cerebral resuscitation.Method Clean male Sprague Dawley rats were randomly divided into sham group and experimental group.Cardiac arrest was induced by asphyxiation and ice-cold 0.5 mol KCl with blood flow cut off for 5 minutes.Animals were resuscitated with external cardiopulmonary resuscitation (CPR),mechanical ventilation,and epinephrine injection.Blood pressure,heart rate,successful ratio of resuscitation after 72 hours, time of cardiac arrest (T_(CA)) and return of spontaneous circulation (T_(ROSC)) were recorded.Neural deficit scores (NDS) and levels of maleic dialdehyde (MDA) in plasma were evaluated at 3,6,12,24,48,72 hours after ROSC.The damage score of cortex was measured by transmission electron microscope examination at 3 hours and 72 hours after ROSC.Results All the rats in experimental group had cardiac arrest rapidly.T_(CA) and T_(ROSC) were (137.3?10.2) seconds and (64.4?9.3) seconds,respectively,while the successful rate of resuscitation was 82.5%.The lowest NDS was at 3 hours after ROSC,while the NDS increased gradually.After CPR,the level of MDA in plasma increased significantly,slightly declined at 72 hours after ROSC,but still significantly higher than before the model.Electron microscope examination of cortex showed neuron slightly,organelle and astrocyte,but became better after 72 hours post ROSC.Conclusions The model of cardiac arrest was easy to establish,and the data provided was accurate,which is useful to study the mechanism of cerebral resuscitation.

Objective To construct the recombinant plasmid pYES6-S and express and purify spike protein of severe acute respiratory syndrome(SANS)coronavirus in Saccharomyces cerevisiae. Methods DNA fragments of SANS coronavirus were obtained by reverse transeription.Four over- lapped fragments of spike protein genes were amplified by polymerase chain reaction(PCR)and ligated into an integral spike protein gene by restriction enzyme digestion.The spike protein gene recombined with pYES6 and cloned into E.coll.The recombinant plasmid pYES6-S was induced and expressed in Saccharomyces cerevisiae(INVScl)by galactose.Results The recombinant plasmid pYES6-S was confirmed that inserted fragment was right in length,direction and base matching by restriction enzyme digestion and sequencing.The purified protein encoded by the whole spike protein gene was about Mr 110?10~3 identified by electrophoresis.Conclusion The whole spike protein gene of SARS coronavirus is cloned into E.coli and the protein is expressed in Saccharomyces cerevisiae successful ly.which can be helpful in SARS vaccine research.