Objective To investigate the short-term treatment effectiveness of the auditory integrative training(AIT) on intelligence, language and gross motor function in children with spastic cerebral palsy (CP).Methods Sixty patients with spastic CP, aged 2 to 4 years old,who had been given all the routine conventional rehabilitation at the Rehabilitation Department of the People's Hospital of Guangxi Zhuang Autonomous Region from January 2011 to December 2013 were randomly divided into AIT treatment group (30 cases) and control group without AIT (30 cases), all patients were investigated by using Gesell Developmental Scales and the Gross Motor Function Measure (GMFM)-66 before and after 3-month therapy, and the changes in the regional cerebral blood flow of children (rCBF) were detected by single photon emission computed tomography (SPECT) before and after treatment.Results The scores of development quotient (DQ) of the treatment group after the AIT in areas of adaptive behavior, personal-social behavior,language(52.44 ± 13.43,52.07 ± 11.57,57.19 ± 6.18) were higher than those of the control group (45.09 ± 11.02,45.86 ± 9.66,53.44 ± 5.49), and the differences between the 2 groups were greatly significant statistically(t =-2.32,-2.26,-2.49, all P ＜ 0.05).There were no significant difference in DQ of gross motor, fine motor and scores on the GMFM-66 in the treatment group before and after treatment [40.40 ± 8.57,49.50 ± 14.20, (55.81 ±8.72) scores vs 44.03 ±11.90,46.34±9.78,(58.81 ±7.86) scores,t=-1.42,1.08,-0.52,all P＞ 0.05].SPECT detected abnormalities in all patients (100.00％) both in the 2 groups, compared with the control group,the rCBF was improved significantly after treatment in the treatment group (86.67％) (x2 =35.49 ,P ＜ 0.05).Conclusions The treatment of AIT can greatly improve the intelligence, language development and the brain function in children with spastic CP.It is an effective adjutant of rehabilitation method for children with spastic CP to improve intelligence and language development, and has less influence on motor function.

The technologies that we call cell micropatterning allow the control of the shape and size of cell adhesion. Combination of micro/nano technology, surface chemistry, electrochemistry and photochemistry enables us to control the adhesion, migration, differentiation of cells and the interactions between different types of cells. These methodologies bring about a new platform for the studies of cell biology. A number of techniques for cell patterning and compares their advantages and disadvantages were reviewed in this article. The applications of cell micropatterning, including those for fundamental studies in cell biology, tissue engineering and cell-based biosensors were also discussed.

The PCR product of PvpA gene was cloned into prokaryotic expression vector pET41a(+) and the recombinant expression vector was then transformed into E.coli DH5a after identified by restriction enzyme digestion and PCR.The positive recombinant plasmid was transformed into E.coli BL21 (D3) and induced to express PvpA protein.The obtained protein was analyzed by SDS-PAGE and Western blotting,purified by Ni-NTA affinity chromatography.The results showed that the purified PvpA fusion protein was obtained successfully.The expressed protein reacted to the high anti-PvpA immune serum from rabbit specially by western blotting.This study would be helpful to established a new diagnostic method for the detection of M.gallisepticum.

Objective To investigate the incidence, risk factors, treatment and clinical outcome of extramedullary EM) relapse following allogeneic hematopoietic stem cell transplantation (alloHSCT), and explore the possible pathogenesis. Methods We retrospectively analyzed the medical records of 164 patients who underwent allogeneic HSCT. The 10 clinical parameters were selected for Cox univariate analysis: gender, age, underlying disease, donor type, disease status at transplant,HLA disparity, acute GVHD, chronic GVHD, EM involvement prior to transplantation and conditioning regimen. Factors that were significant at the 0. 10 level on univariate analysis were evaluated by multivariate analysis using a Cox regression. The therapeutic options for EM relapse included local radiation, surgical removal, chemotherapy, donor lymphocyte infusion (DLI), second HSCT. Results 164 recipients had sustained engraftment. EM relapse occurred in 9 patients(5.5 %), with a median time to EM relapse of 7.5 months (2.3 to 42.6 months). Ninety-fourpatients (57. 3 % ) developed acute GVHD and 83 (50. 6 % ) chronic GVHD respectively. Four patients died of EM relapse. The following factors were associated with an increased risk of EM relapse by univariate analysis: gender, donor type, disease status at transplant, chronic GVHD, EM involvement prior to transplantation. Only advanced stage of the disease (P＜ 0. 05), absence of chronic GVHD (P＜0. 01) and EM involvement prior to transplantation (P＜0. 01) were identified as being significantly associated with the occurrence of EM relapse by multivariate analysis using a Cox regression. Conclusion Many factors may be involved in the pathogenic mechanism of EM relapse,and among them, immune escape might play a major role. Advanced stage of the disease, absence ofchronic GVHD and EM involvement prior to transplantation were independently associated with an increased risk of EM relapse. EM relapse frequently followed by bone marrow involvement has poor prognosis, and therefore, prevention of leukemic cells spreading from EM sites to bone marrow is vital for long-term survival.

The chemical structures of mequindox related metabolites in chicken plasma had been investigated using high performance liquid chromatography combined with linear ion trap quadrupole(LC-ESI/LTQ) and high performance liquid chromatography combined with ion trap-time of flight-mass spectrometry (LC-ESI/IT-TOF).Samples were separated by Hypersil BDS C_(18) and symmetry Shield columns, respectively, and 0.01% formic acid aqueous(A) and methanol(B) were used as mobile phase with gradient elution.Electros pray ionization mass spectrometric(ESI) source was used and operated in positive ion mode.When chickens were orally administered with mequindox at dosage of 20 mg/kg, blood samples were collected from the brachi al vein.Mequindox and its metabolites were extracted by the mixture of acetonitrile and acetoacetate (3:2, V/V).After solvent evaporated, the residue was dissolved in 30% methanol aqueous and the solution was detected by LC/IT-TOF MS and LC-ESI/LTQ.The molecule weight from LC-ESI/IT-TOF was analyzed by software Shimadzu's Composition and the mass chromatogram from LC-ESI/LTQ was analyzed by software Xcalibur 2.0.7.According to the molecular weight and MS~n data, referring the metabolic reaction rules, five chemical structures of mequindox related metabolites in chicken plasma were identified.Metabolites (M1-M4) were synthesized to verify the structure of metabolites.The metabolites are 3-methyl-2-(1-hydroxy) ethyl-qui-noxaline-N~1,N~4-dioxide(Ml), 3-methyl-2-(1-hydroxy) ethyl-quinoxaline-N~4-oxide(M2), 3-methyl-2-acetyl-quinoxaline-N~4-oxide, 3-methyl-2-acetyl-quinoxaline (M4), 3-hydroxymethyl-2-(1-hydroxy) ethyl-quinoxa-line-N~1,N~4-dioxide (M5).

Objective To further investigate inhibitory mechanism(s) of resveratrol on varicellazoster virus (VZV) in vitro with our previously generated reporter cell line MV9G.Methods Cell-free VZVs were directly inoculated onto MV9G cells (CFVs direct-infection) or cell-associated VZVs wereco-cultured with MV9G cells (CAVs co-culture) to activate expression of reporter gene firefly luciferase in MV9G cells.Resveratrol was added before or after virus infection,roles of resveratrolon direct inactivation,on viral attachment to and penetration into MV9G cells,on intracellular viral replication and its IC50,inhibitorytime points and reversibility were assayed by comparing the luciferase activities reduction by resveratrol.Thereductions of VZV IE62 mRNA copies and IE62-antibody positive cells by resveratrol were further assayed.Results ATPs contents of MV9G cells in the presence of resveratrol over 30.0 μg/ml were concentrationdependently reduced,the CD50 of which was around 60.3 μg/ml.CFVs were premixed with 25.0 μg/ml resveratrol andincubated at 37℃ waterbath for two hours and then directly inoculated onto MV9G cells,luciferases activated by resveratrol-treated CFVs were reduced to around half of the untreated controls.MV9G cells were pre-incubated with resveratrol at 37℃ for 2 h and then directly infected with CFVs at 37℃ for another 2 h,the CFVs-activated luciferase was concentration-dependently reduced,but no big change was observed in those pre-incubated at 4℃.MV9G cells were co-cultured with CAVs in the presence of resvertrolfor 72 h,the CAVs-activated luciferases were markedly reduced in a concentration-dependent manner,the IC50 of which was around 8.7 μgml.Resveratrol was added in CAVs co-culture at 1,3,6,9,12,24,30,and 36 h post infection,the CAVs-activated luciferase in those resveratrol was added at 3,6,9,12,and 24 h post infection were significantly higher than those of controls.Resveratrol was withdrawn from CAVs coculture media,the CAVs-activated luciferases after withdrawal were significantly higher than those before,especially in those withdrswn at 24 and 72 h post infection.The IE62 mRNA levels shown by cDNA copiesdetected with SYBR Green RT-PCR and IE62 positive cells shown by monoclonal anti-IE62 antibody of thevirus-infected cells treated with resveratrol were significantly reduced with increase of incubation time withresveratrol.Conclusion Resveratrol was cytotoxic to MV9G cells,and the maximum resistant concentrationon MV9G cells was around 30.0 μg/ml,the CD50 of which was around 60.3 μg/ml.Non-cytotoxic resveratrol partly inactivated CFVs,inhibited viral penetration into rather than attachment to MV9G cells.Resveratrol inhibited CAVs' intmcellular replication strongly but reversibly in a concentration-dependent manner,the IC50 of which was around 8.7 μ/ml.The inhibition of resveratrol on VZV in vitro might be through suppression of IE62 gene transcription and expression in the early stage of infection.

BACKGROUND:Endothelial progenitor cells have good prospects for clinical application;especial y as seed cells, they are involved in construction of tissue engineered blood vessels and disease treatment. OBJECTIVE:To investigate the biological characteristics of endothelial progenitor cells derived from granulocyte colony-stimulating factor (G-CSF) mobilized peripheral blood. METHODS:Mononuclear cells from G-CSF mobilized peripheral blood (n=9) and normal adult peripheral blood (n=8) were obtained and cultured in expanded medium. The immunophenotype of endothelial progenitor cells was investigated by FACS and immunohistochemistry. Real-time PCR was used to detect the expression of different cytokines. Proliferation and adhesion ability of endothelial progenitor cells were detected by MTT assay and adhesion assay. Moreover, the abilities of vasculogenesis in vitro and Dil-labeled acetylated low density lipoprotein uptake were detected. The acquired cells were seeded onto the basilar membrane gel containing vascular endothelial growth factor and basic fibroblast growth factor to induce angiogenesis. RESULTS AND CONCLUSION:Endothelial progenitor cells derived from G-CSF mobilized peripheral blood were similar to those from normal adult peripheral blood in phenotype and morphology. FACSs and immunohistochemistry showed that endothelial progenitor cells were positive for endothelial cellmarkers, such as CD31, vWF, CD34, FLK-1, VE-Cadherin and CD133. In addition, the expression of vascular endothelial growth factor and stromal cel-derived factor 1 were significantly increased in G-CSF mobilized endothelial progenitor cells. Moreover, endothelial progenitor cells derived from two sources possessed the abilities of angiogenesis in vitro and Dil-labeled acetylated low density lipoprotein uptake. But, the number and the proliferation ability of endothelial progenitor cells derived from G-CSF mobilized peripheral blood were better than that of endothelial progenitor cells derived from normal adult peripheral blood. These findings indicate that there is a population of cells with characteristics of endothelial progenitor cells in G-CSF mobilized peripheral blood. They possess the abilities of vasculogenesis in vitro. Moreover, compared to endothelial progenitor cells derived from normal adult peripheral blood, we could obtain more endothelial progenitor cells in G-CSF mobilized peripheral blood and those cells show better proliferation ability.

Objective To investigate the effect of extracellular histones (EH) on intestinal mucosal barrier function in mice and the correlation of EH with the pathogenesis of sepsis.Methods Twenty male C57BL/6 mice were assigned into experiment group (n =10) and control group (n =10) according to the random number table.Same dose (50 mg/kg) of EH and saline were administered through the caudal vein of mice in experiment and control groups respectively.Blood and intestinal samples in each group were collected 3 h after the administration.Morphology of intestinal mucosal tissue was detected by light scope and transmission electron microscope.Expressions of tight junction related proteins (ZO-1,Occludin and Claudin-1) were detected by western blot.Plasma levels of diamine oxidase (DAO) and intestinal fatty acid binding protein (I-FABP) were determined by ELISA method.Plasma level of endotoxin (ET) was determined by limulus test.Results Under transmission electron microscope,experiment group showed disorganized microvilli of intestinal epithelial cells with partially twisted,broken and lost,unclear tight junctions,and widened cellular space.Under light scope,experiment group showed substantial inflammatory cell infiltration in the intestinal wall,disorganized intestinal villi,edema and hemorrhage of mucosa and submucosa,and edematous goblet cells.Experimental versus control group showed significant reduction in levels of Claudin-1 (0.587 7 ±0.060 6 vs.0.677 2 ±0.038 3),Occludin (0.1277±0.0857vs.0.4306±0.0869) and ZO-1 (0.393 3±0.080 8 vs.0.812 8± 0.096 3) (P ＜ 0.05).Experimental versus control group showed significantly up-regulated plasma levels of DAO [(1.61 ±0.20) U/ml vs.(0.69 ± 0.15) U/ml],I-FABP [(548.5 ± 36.8) EU/ml vs.(178.8±26.9) EU/ml] andET [(0.182±0.076) EU/mlvs.(0.091 ±0.029) EU/ml](P＜0.05).Conclusion EH can obviously impair the integrity of intestinal mucosal barrier in mice and hence induce endotoxin translocation.

Objective:To explore influence of thrombus aspiration combined bivalirudin during emergency PCI on my- ocardial tissue perfusion and clinical prognosis in patients with acute ST elevation myocardial infarction (STEMI). Methods:A total of 102 patients with acute STEMI,who were confirmed with thrombus burden by CAG in our hos- pital from Jan 2012 to Jun 2014,were selected.According to random number table,they were randomly divided into thrombus aspiration + bivalirudin group (n=52,thrombus aspiration group)and heparin group (n=50,routine PCI group).TIMI blood flow grade 3 rate,TIMI myocardial perfusion grade (TMPG)after PCI,ST segment re- gression rate 2h after PCI,peak value and peak time of cTnI after PCI,LVEF,LVEDd,incidence rates of bleeding and major adverse cardiovascular events (MACE)on one week and one month after PCI were compared between two groups.Results:Compared with routine PCI group,there were significant rise in postoperative TMPG grade 3 rate (56.00% vs.88.46%),TIMI grade 3 rate (58.00% vs.88.46%)and ST segment regression rate (52.00% vs. 76.92%)in thrombus aspiration group,P<0.05 or <0.01. Compared with routine PCI group one month after PCI,there was significant rise in LVEF [(53.76±5.24)% vs.(57.95±5.51)%],and significant reductions in LVEDd [(53.70±3.39)mm vs.(50.63±1.24)mm],peak value [(16.00±4.28)μg/L vs.(13.81±4.00)μg/L]and peak time [(14.00±2.80)h vs.(13.00±2.23)h]of cTnI in thrombus aspiration group,P<0.05 or <0.01. Incidence rate of mild bleeding in thrombus aspiration group was significantly lower than that of routine PCI group (1.9% vs.16.0%),P<0.05,but there was no significant difference in incidence rate of MACE between two groups,P>0.05. Conclusion:Thrombus aspiration combined bivalirudin during emergency PCI is safe and fea- sible for acute STEMI patients,it can effectively reduce incidence rate of bleeding,remove coronary thrombus,im- prove myocardial tissue perfusion and doesn't increase incidence rate of MACE.

Objective:To observe influence of up‐regulated expression of myocardial heat shock protein (HSP) 70 in‐duced by heat stress on myocardial calcium‐activated potassium channel (KCa ) 3.1 expression in rabbits with atrial fibrillation (AF) caused by rapid atrial pacing (RAP) .Methods :A total of 24 New Zealand white rabbits were ran‐domly divided into sham operation group (n=8 ,only implant electrode without pacing ) ,pacing group (n=8 ,right atrium (RA) received RAP at 600 times/min for 6h) and heat stress pacing group (heat stress group ,n=8 ,received heat stress preconditioning ,then the same RAP as pacing group ) .Results:Compared with sham operation group and pacing group ,there were significant up‐regulation of HSP70 mRNA and protein expression in different sites of heart [HSP70 protein ,left atrium (LA):(39.00 ± 3.21) vs .(39.75 ± 2.82) vs .(69.75 ± 3.45) ,RA: (38.38 ± 2.92) vs .(39.50 ± 3.89) vs .(69.00 ± 2.93) ,left atrial appendage (LAA):(37.75 ± 3.28) vs .(39.00 ± 3.89) vs . (68.63 ± 3.23) ,right atrial appendage (RAA): (37.00 ± 3.85) vs .(38.38 ± 3.74) vs .(68.75 ± 2.82)] in heat stress group , P<0. 01 all ,but there were no significant difference between pacing group and sham operation group , P>0.05 ;compared with pacing group with down‐regulation of KCa3.1 mRNA and protein expressions ,there were significant up‐regulation of KCa3.1 mRNA and protein expressions in different sites of heart [KCa3.1 protein ,LA:(21.25 ± 1.67) vs .(24.00 ± 2.62) ,RA :(21.13 ± 1.96) vs .(23.75 ± 1.83) ,LAA :(21.00 ± 2.07) vs .(23.75 ± 1.67) ,RAA:(20.88 ± 2.03) vs .(23.50 ± 2.45)] in heat stress group ,P<0.05 all ,and there were no significant difference between heat stress group and sham operation group , P>0. 05. Conclusion:Heat stress may induce up‐regulated expression of myocardial HSP 70 of myocardium ,and HSP 70 may inhibit down‐regulation of KCa 3. 1 mR‐NA and protein expressions in rabbits with atrial fibrillation.