Objective To evaluate the regulatory effect of OX40 co-stimulatory signal on the expression of Foxp3 in inductive regulatory T cells (iTreg) in vitro.Method CD4+ CD25+ naive T cells were isolated from C57BL/6 mouse lymphocyte suspension by MASC CD4+ CD25+ regulatory T cell isolation kit.Inductive Tregs were generated by stimulation of naive T cells in the presence of transforming growth factor beta (TGFβ1),anti-CD3,anti-CD28 and IL-2.The regulatory effect on iTregs was shown by use of OX40 stimulation monoclonal antibody (OX86) or control antibody.Using flow cytometric analysis (FACS),we examined the antibody-based identification of Tregs surface markers CD4 and CD25,along with the intracellular activation marker FoxP3.Results The ratio of CD4+ CD25+ nTregs isolated from mouse lymphatic node was (5.0 ± 0.4)％ vs.(71.8 ± 13.4)％ of TGFβ1-driven iTregs.The ratio of CD4+ CD25+ Tregs was (80.0 ± 1.6) ％ in OX40 stimulation McAb group vs.(86.0 ± 1.4)％ in control antibody group.Furthermore,the expression of Foxp3 was (59.2 ± 0.7) ％ in OX40 stimulation McAb group vs.(70.0 ± 0.8) ％ in control antibody group (P＜0.05).Conclusion TGFβ1-dependent protocol may induce the conversion of naive CD4+ T cells into CD25+ Foxp3+ iTregs.OX40 stimulation can down-regulate the expression of Foxp3 in CD4+ CD25 + iTreg significantly.Thus OX40 molecular may become an attractive target in Tregs-induced transplant tolerance.Further study should be performed to increase the suppressive activity of iTregs through blockade of OX40 signal.

Objective To evaluate the therapeutic effect of esophageal intraluminal stents with 125I seeds based on survival time and quality of life (QOL) of patients with advanced esophageal cancer.Methods A total of 37 patients with advanced esophageal cancer who underwent placement of esophageal 125I irradiation stent were followed up.The QOL was evaluated by QLQ-C30 and QLQ-OES18 questionnaires before the procedure, and 1 week and 3 months after.Results The mean survival time of 37 patients was 28 weeks, with 3-month and 6-month survival rates at 94.6% (35/37) and 51.4% (19/37), respectively.Compared with the baseline, the scores of emotional functioning(EF), cognitive functioning(CF),global quality of life(QL), fatigue(FA), pain(PA), financial difficulties(FI) and pain of QLQ-OES18(OESPA) assessed 1 week after stent placement increased (P<0.05) significantly and dysphapia of QLQ-OES18(OESDYS), eating of QLQ-OES18(OESEAT), trouble swallowing saliva of QLQ-OES18(OESSV) and choked when swallowing of QLQ-OES18(OESCH) decreased significantly (P<0.05).At 3 months after stent placement, the scores of FI increased significantly (P<0.05), and those of nausea or vomiting (NV), OESDYS,OESEAT, OESSV and OESCH decreased significantly (P<0.05).The scores of FI,OESDYS, OESCH, role functioning(RF), EF,CF,FA and OESPA at 3 months were significantly different from those at 1 week after the procedure (P<0.05).Other variables at 1 week and 3 months after the procedure were not different from those before (P>0.05).Conclusion Implantation of 125I seeds combined with esophageal intraluminal stents may prolong survival time and improve QOL of patients with advanced esophageal cancer.

Objective To compare volumetric‐modulated arc therapy(VMAT) with intensity‐modulated radiation therapy (IMRT) for brain metastases with regard to the dosimetric character .Methods Sixty patients who were diagnosed with brain me‐tastases were included in this study .The target area received two dose levels using late addition amount technique ,WBRT (30 Gy/10 F) with following addition (20 Gy/10 F) to 59 Gy .For a fair comparison ,VMAT and IMRT treatment plans were respectively designed for every patient with the same dosimetric constraints .Dosimetric comparisons between VMAT and IMRT plans were ana‐lyzed to evaluate :target coverage and homogeneity ,conformity of PTV ;sparing of OARs ;monitor units (MUs) .Results Two treatment plans all reached the treatment need .When compared with IMRT ,there was no significant difference in Dmean of eyeball , len ,optic never ,visual chiasma ,parotid ,brain stem ,and external auditory canal of VMAT (P>0 .05) .The Dmax of eyeball ,len ,pa‐rotid ,and external auditory canal of VMAT were lower than that in IMRT group (P<0 .05) .The VMAT group has the less MUs (P=0 .017) and less treatment time .Conclusion VMAT can reach the big‐dose radiotherapy need on brain metastases clinically . There are no significant diffference between VMAT and IMRT on Dmax ,Dmean ,CI ,and HI .The Dmax of eyeball ,len ,parotid ,and external auditory canal of VMAT were lower than that in IMRT group .The VMAT can reduce the radiotherapy time .

OBJECTIVETo compare the effect,safety,and advantage of flexible fixation with paperboard and pad versus short leg plaster in treating the fifth metatarsal base fracture,and establish the standard of diagnosis and treatment of the fifth metatarsal base fractures in flexible fixation with paperboard and pad.

METHODSFrom June 2010 to March 2013,59 patients with the fifth metatarsal base fracture were treated with paperboard and pad fixation or short leg plaster. Patients were enrolled and divided into paperboard and pad treatment group (paperboard group) and short leg plaster treatment group (plaster group) randomly according to the random number table. In paperboard group,there were 29 cases including 9 males and 20 females with an average age of (51.79±11.40) years old; the average course of injury was (11.59±6.58) hours. In plaster group, there were 30 cases including 9 males and 21 females with an average age of (52.13+17.34) years old ;the average course of injury was (11.03±7.06) hours. According to whether the fracture line across the articular surface, in paperboard group there were 14 cases of type A,15 of type B; in plaster group,16 of type A, 14 of type B. According to the degree of dislocation,in paperboard group there were 16 cases of degree I ,13 of degree II ; in plaster group,20 were degree I ,10 were degree II. Fracture was restored according to the type in manual. Patients in paperboard group were treated with paperboard and pad, and patients in plaster group were treated with short leg plaster. Fracture was fixed for 4 to 6 weeks according to fracture healing. On the 2nd, 4th,6th, 8th week and 3rd, 6th month after fixation, patients were followed up, and the foot function score was used to evaluate the function of injured foot. X-ray of injured foot was taken on the 2nd, 4th, 6th and 8th week were used to assess fracture healing.

RESULTSAll patients got complete follow-up. The X-ray result showed that all fracture reached at clinical healing on the 8th week after fixation without skin ulcer,nonunion and displacement of fracture. From the 4th to 8th week after fixation, paperboard group had a higher X-ray score than plaster group, but the difference between two groups had no statistically significance. Repeated analysis result showed that there was interact at different time point and between groups,the difference had statistically significance (P<0.01). The foot function score showed that at all time point, paperboard group had a higher score than plaster group, and on the 2nd, 4th, and 6th week, it had statistically significant difference(P<0.01) between two groups. On the 6th months after fixation,the excellent and good rate of paperboard group was 93.10%, higher than that of plaster group, which was 86.67%. But it had no statistically difference(P=0.483) between two groups.

CONCLUSIONUsing paperboard and pad fixation to treat the fifth metatarsal base fracture has the advantage of simplicity operating,reliable fixation, satisfactory effects, easily obtainable material.

Adult ; Aged ; Casts, Surgical ; Female ; Foot Injuries ; physiopathology ; surgery ; Fracture Fixation ; instrumentation ; methods ; Fracture Healing ; Fractures, Bone ; physiopathology ; surgery ; Humans ; Male ; Metatarsal Bones ; injuries ; physiopathology ; surgery ; Middle Aged

Objective:To investigate the effect of afatinib, a tyrosine kinase inhibitor, on the proliferation, cell cycle, and apoptosis of human breast cell lines, and compare its effects with those of gefitinib. Methods:Three human breast cell lines, MCF-7, T47D, and MDA-MB-231, were cultured as cell models. A methyl thiazolyl tetrazolium assay was utilized to measure cell viability. Flow cytometer was used to analyze the cell cycle arrest (PI staining) and apoptosis rates (Annexin-V/PI staining). The protein expression was detected by Western blot analysis. Results:The proliferation of three human breast cell lines was significantly inhibited by afatinib, and the IC50 levels of MCF-7, T47D, and MDA-MB-231 were 0.101, 0.141, and 0.887μmol/L, respectively. The G0/G1 phase cell ratio increased con-siderably 24 h after afatinib was added to T47D or MDA-MB-231. The cell apoptosis rate also increased in the two cell lines (88.9%and 58.1%). The cleavage of apoptosis pathway proteins PARP and caspase-3 was also promoted by afatinib. Phosphorylation of EGFR was significantly inhibited by afatinib in the MDA-MB-231 cell line. Finally, the inhibition effect of afatinib was stronger than that of gefi-tinib. Conclusion: Afatinib could significantly inhibit the proliferation of breast cancer cells and promote apoptosis. The effect was dose-dependent. Afatinib was a more effective tyrosine kinase inhibitor as compared with gefitinib.

Background Lasein situ keratomileusi(LASIK) imainstream surgery forefractive correction,and femtosecond laseimuch often used to create thin corneal flap.The measuremenof OPTOVUE RTVue-100 OCto flap and stromal bed thicknesseofferuseful basifoLASIK.Ican be used in measuring the thicknesand shape of the corneal flap.Buthe study on the comparison of flap thicknesbetween WavelighFS200 femtosecond laseand MoriM2 microkeratome 90 μm-knife (Mori90 microkeratome) LASIK by OCilack.Objective The aim of thitrial wato compare the featureof corneal flapcreated by the WavelighFS200 femtosecond laseand Mori90 microkeratome.Methodpiloand prospective study wadesigned.Written informed consenwaobtained from each patienprioto LASIK.Sixty righeyeof 60 patientwith myopiomyopiastigmatism were enrolled in thiclinical trial.The patientwere randomized into the FS200 femtosecond lasegroup and Mori90 microkeratome group with matching demography.RTVue OCwaused to measure flap thicknesusing 10 settingon the 60 eye1 month afteoperation.The featureof the LASIK flapwere analyzed based on the measuring outcomes.ResultThe central flap thickneswa(112±3) μm and the mean flap thickneswa(112 ±3) μm in the FS200 femtosecond lasegroup,which wasignificanlowethan the central flap thicknesa(121±7) μm and the mean flap thicknesa(128±11) μm in the Mori90 microkeratome group respectively (P=0.031,0.030).Corneal flapin the FS200 femtosecond lasegroup showed flashape and thain the Mori90 microkeratome group wameniscushape.The central flap thickneswanoevidently differenfrom thaof peripheral thicknesin the FS200 femtosecond lasegroup (P =0.320).However,in the Mori90 microkeratome group,the central flap thickneswaobviously thinnethan thain the peripheral thicknes(P=0.038).The mean deviation between the actual and predicted flap thicknes(110 μm) wa(3±4)μm in the FS200 femtosecond lasegroup and (17±10) μm in the Mori90 microkeratome group,showing significandifference between them (P =0.009).ConclusionRTVue OCdeterminethathe shape of flapcreated by the FS200 femtosecond laseimore uniform and closeto the expected thicknesof 110 μm than the onecreated by the Mori90 microkeratome.OPTOVUE RTVue-100 OCiuseful tool to evaluate the flap shape and thicknesafteLASIK.

Objective To investigate the prevalence,genotype and epidemiology of plasmid- mediated AmpC enzyme of Escherichia coli and Klebsiella pneumoniae.Methods A total of 67 clinical isolates of nonrepetitive cefoxitin-resistant Escherichia coli and Klebsiella pneumoniae collected by Fuzhou General Hospital and Fujian Provincial Hospital during a period of Sept.2004 to Mar.2005 were detected by three-dimensional extract test for AmpC enzyme,and PCR for AmpC enzyme and other ?-lactamase gene amplification and DNA sequencing were carried out for genotype of ?-lactamase.Plasmid transformation experiment was used to study the transfer of cefoxitin resistance.The homology of the isolates was determined by ERIC-PCR fingerprinting.Results At two hospitals in Fuzhou,the prevalence of plasmid-mediated AmpC enzyme among cefoxitin-resistant Escherichia coli and Klebsiella pneumoniae were 16.7% and 10.5%, 8.0% and 0,respectively.Two isolates of Klebsiella pneumoniae produced DHA-1 plasmid-mediated AmpC enzyme,and 4 isolates of Escherichia cob and one strain of Escherichia coli produced CMY-2 and CMY-22 plasmid-mediated AmpC enzyme respectively.Furthermore,5 strains of Escherichia coli with CMY AmpC enzyme were also found simuhaneously to produce TEM-144,CTX-M-27,CTX-M-14 and TEM-1 ?-lactamase respectively.Three strains of Escherichia coli and one isolate of Klebsiella pneumoniae could transfer cefoxitin resistance to acceptant bacillus.ERIC-PCR fingerprinting reveals 2 strains of Klebsiella pneumoniae came from same clone,but 5 strains of Escherichia coli came from different clones.Conclusions The clinical isolates of Klebsiella pneumoniae producing DHA-1 plasmid-mediated AmpC enzyme and Escherichia coli producing CMY-2,CMY-22 plasmid-mediated AmpC enzyme are found in Fuzhou.CMY-22 AmpC enzyme and TEM-144 ?-lactamase are the first reported in the world,GenBank accession number: DO256079,DO256080

AIMTo clone the gene of staphylococcal enterotoxin C2 and express it in the form of a soluble fusion protein in E. coli. Then the activation of SEC2 on mice lymphocyte and its lethal effects on tumor cells were studied.

METHODSStaphylococcus aureus SEC2 gene was cloned into GST gene fusion vector pGEX-4T-1. The resultant plasmid pGEX-4T-SEC2 was used to transform E. coli BL21, where the GST-SEC2 fusion protein was expressed efficiently. The rSEC2 protein was purified with Glutathione Sepharose 4B affinity column and digested with thrombin. The in vitro culture system was utilized to observe the activation of the SEC2 on mice lymphocyte and the lethal effects on tumor cells of the activated mice lymphocyte.

RESULTSThe proper gene of SEC2 was cloned and purified rSEC2 was obtained. The MTT results indicated that rSEC2 have strong ability to stimulate mice lymphocyte to proliferate with a dose-dependent manner. With the proliferation of mice splenic lymphocyte, rSEC2 has a strong lethal effect on tumor cells B16, K562 and K562-AD.

CONCLUSIONIn this study, the gene of SEC2 was cloned and the rSEC2 protein was obtained, which had strong lethal effect on tumor cells B16, K562 and K562-AD.

Animals ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; Cloning, Molecular ; Enterotoxins ; genetics ; metabolism ; pharmacology ; Escherichia coli ; genetics ; metabolism ; Female ; Genetic Vectors ; Glutathione Transferase ; genetics ; Lymphocyte Activation ; drug effects ; Lymphocytes ; cytology ; immunology ; Male ; Mice ; Mice, Inbred ICR ; Recombinant Fusion Proteins ; genetics ; metabolism ; pharmacology ; Spleen ; cytology ; Transfection

Animals ; Captopril ; pharmacology ; Heart Failure ; metabolism ; Male ; Myocytes, Cardiac ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Sprague-Dawley ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo investigate whether the mitochondrial ATP-sensitive potassium channel (mitoK(ATP)) opener diazoxide as an additive to cardioplegia solution could enhance myocardial protection during hypothermic preservation of the rat heart.

METHODSThe Langendorff model of isolated rat heart was used. After equilibrium, the hearts were stored in Celsior cardioplegia solution at 4 degree with or without supplement of diazoxide for 3 or 8 h followed by 60 minutes reperfusion. The recovery of cardiac contractile function, myocardial enzyme leakage in the coronary effluent, and myocardial water content were determined. The myocardial ultrastructure was also observed.

RESULT(1) Treatment of diazoxide improved the recovery of left ventricular developed pressure and decreased the leakage of myocardial enzymes, lactate dehydrogenase (LDH) and creatine kinase (CK), at the 2nd and 4th minute of reperfusion of rat heart after hypothermic preservation for 3 h. (2) After hypothermic preservation for 8 h, diazoxide improved the recovery of left ventricular developed pressure and decreased the leakage of myocardial enzymes (LDH, CK and glutamic oxalic transaminase) during reperfusion. Moreover, left ventricular end-diastolic pressure was significantly lower in diazoxide-treated hearts than that of hearts in Celsior solution. (3) Diazoxide significantly decreased the water content of myocardium and increased coronary flow of the hearts compared with those in control after hypothermic preservation for 8 h. (4) Impairment of myocardial ultrastructure after 8 h hypothermic preservation was alleviated in hearts treated with 30 mol/L diazoxide. (5) The cardiac effects of 30 mol/L diazoxide were attenuated by a mitoK(ATP) blocker 5-hydroxydecanoate (100 micromol/L).

CONCLUSIONDiazoxide as a supplementation in cardioplegia solution could enhance myocardial protection during hypothermic heart preservation via opening of mitochondrial K(ATP) channel.

Animals ; Cardioplegic Solutions ; Cryopreservation ; Diazoxide ; pharmacology ; Heart ; Male ; Organ Preservation ; Organ Preservation Solutions ; pharmacology ; Potassium Channels ; drug effects ; Rats ; Rats, Sprague-Dawley