0.05).Our data also showed no significant association between the genotypes and the severity of the disease.One-way ANOVA showed that BDNF genotype had no association to the age of onset for developing AD.Conclusions Our results indicate that Va166Met SNP in BDNF gene is not associated with AD.

Objective: To investigate the possibility of adipose tissue mesenchymal stem cell-derived microvesicles (ADSC-MVs) to improve the retention volume of fat transplantation. Methods: Human adipose tissue was obtained from 5 healthy female patients aged from 20 to 30 years, who came to the hospital for abdominal liposuction. Adipose tissue mesenchymal stem cells (ADSCs) were acquired by collagenase enzymatic hydrolysis. ADSC-MVs were isolated from the supernatant of cultured ADSCs through ultra-centrifugation, and characterized by transmission electron microscope and PKH26 staining. Sixteen BALB/c-nu nude mice were randomly divided into 2 groups (n=8, mice/group) by random number table method. In EV group, the mice were subcutaneously injected 0.35 ml human fat, with 7 μg ADSC-EVs (final concentration is 20 μg/ml), while 0.35 ml human fat were in blank group. The mice were sacrificed and the grafts were harvested at 1 month and 3 months after transplantation. Weight and volume of transplanted fat were tested. Morphology of adipocytes, fibrosis and necrosis of fat tissue were confirmed by HE staining. Blood vessel density were accessed by CD31 staining.SPSS 13.0 was used for statistical analysis and Student-t test was applied to compare the data from two groups. Results: One month after fat transplantation, the volume of fat grafts in blank group was lower than that in MV group. Besides, lipoliquefaction can be observed in fat grafts of blank group. Fat grafts in MV group had a higher volume and littler necrosis and lipolique faction. Three months after grafting, the volume of fat grafts in blank group [(0.057±0.009) ml] was significantly lower than that in MV group [(0.103±0.015) ml], t=8.117, P<0.001. MV group had an increased number of vessels (29.6±4.0/field) compared with grafts from the Blank group (23.0±2.4/field), t=2.825, P=0.022. Histological analysis revealed that the fat grafts in MV group consisted less fat necrosis and fibrosis than control group. Conclusions ADSC-MVs can improve the retention volume of fat transplantation.

Objective To explore the feasibility of recently developed nucleofection method in delivering plamid DNA directly into the nucleus for the introduction of a plasmid encoding enhanced green fluorescent protein (EGFP) into primary bone marrow stromal cells (BMSCs) of rabbit. Methods Rabbit BMSCs were harvested by means of density gradient centrifugation following a thighbone puncture. The primary BMSCs were cultured and either transfected with pEGFP-C2 by nucleofector technology (as EGFP group) or uninfected (as control) in vitro. Compared with the control, the cellular viability, adhesive rates and the growth curves of the labeled cells were respectively analyzed. Transfection efficiencies were evaluated through the detection of EGFP expression. Results EGFP were successfully expressed 24 h after nucleofection. Similar morphological development, adhesive rates and growth curves were found in the 2 groups. The positive EGFP expression was enhanced gradually alone with the prolonged culture time, and showed the strongest 6 d after marked, with about 47.8% of EGFP-positive cells in the total BMSCs. The EGFP did not attenuate even 1 month after the marking. Conclusion Neuclofection of pEGFP-C2 shows no significant effect on the proliferation of rabbit BMSCs. EGFP plays an important role in stable gene marking of rabbit BMSCs. Nucleofection is an efficient nonviral gene transfer method for the introduction of genes into primary rabbit BMSCs.

Objective To offered some prophase works by preparing Neurocan protein, antiserum, and assaying their characteristics, in order to construct the isopropy-β-D-thiogalactoside (IPTG)-participated DNA vaccine, which can neutralize the inhibitors in the injured CNS following the immune administration and then promote the nerve regeneration. Methods Neurocan gene was syntbetized with HisTag label in beginning and enzyme-cut sites at amphi- of the sequence. The prokaryotic expression plasmid, PET30a-Neurocan, was constructed as usual, converted into the Escherichia coli, and induced by IPTG to express positively. The interest protein was identified by SDS-PAGE and Western blot respectively. The preimmune serum was as the negative control during the ELISA assay of antiserum valency. The immune serum was as the first antibody, and the goat-anti-rabbit labeling with alkaline phosphatase (AP) was employed as the second one. Coloration was with NBT/BCIP method. Results The correct sequence of the synthetic Neurocan gene was clearly showed by identification with enzyme-cut, PCR and sequencing. The Neurocan protein expressed by prokaryotic showed its molecular weigh as 55 000 following the SDS-PAGE identification, and it could specifically bind with anti-HisTag, which implied the interesting protein just as the expression product of Neurocan gene. The valency of antiserum was shown by ELISA as 1:1 000 000, the purpose strap of which was confirmed by Western blot. Conclusions Neurocan protein could be successfully expressed in prokaryotic, the antibody of which could be specifically obtained by immune administration to the rabbit. The Neuroncan antibody could bind with the Neurocan protein specifically.

Objective To study the effects of environmental factors,including high temperature,high humidity and low temprtature,on the quality of whole blood.Methods Fresh blood was collected from 9 blood donors and divided into 40 parts.One group that comprised 20 samples was used to assess the effects of storage at high temperature and high humidity (0,3,6,12 and 24 h),and the other group(20 samples)was used to determine the effects of low temprtature(0,1,2 and 3 h).The serum free hemoglobin(FHb)concentration, plasma prothrombin time(PT), activated partial thromboplastin time(APTT),thrombin time(TT)and fibrinogen(FIB)were measured and analyzed.Results The high temperature(27-37℃)and humidity(60%-90%RH)had no effect on FHb, PT, APTT, TT or FIB of whole blood. The low temperature(0-13℃and -18--20℃)had some effect on the quality of whole blood.The FHb of test group was higher than that of control group.There was no significant difference in PT, APTT, TT and FIB between the two groups.Conclusion Under high temperature and humidity conditions, 24 hours of whole blood placement is feasible. Under low temperature or cold conditions,short-term whole blood placement is feasible in a tent with heating facilities.

Objective To analyze the prevalence rate of diabetic nephropathy (DN) and the related factors on DN among type 2 diabetic patients.Methods A total number of 1758 type 2 diabetic patients who were hospitalized in the Beijing Hospital from 2003 to 2010 were analyzed retrospectively.Three groups were divided according to the rate of urinary albumin excretion (UAER).Patients whose UAER＜20 μg/min belonged to normal albuminuria (NA) group.The ones whose UAER from 20 to 200 μ g/min belonged to microalbuminuria (MA) group,and the others whose UAER≥200 μg/min belonged to large albuminuia (LA) group.The clinical characteristics were then compared.The related factors of DN were analyzed.Results (1)There were 1246 patients in NA group,408 patients in MA group,and 104 patients in LA group.The constituent ratio of nephropathy was 29.1％ (2) The ages of NA group,MA group and LA group were (59.87± 12.77,62.52± 12.74,64.44 ± 12.74) years old,respectively,with body mass index ( BMI ) as (24.90 ± 3.42,25.53 ± 4.00,25.53 ± 3.91 )kg/m2 respectively; duration of diabetes as (8.39 ± 7.12,10.77 ± 8.02,12.84 ± 7.97)years; systolic blood pressure (SBP) as (133.42 ± 18.19,142.72 ± 20.21,151.12 ± 21.91 )mm Hg;diastolic blood pressure as (78.75 ± 10.66,80.79 ±12.21,83.33 ±13.61 )mm Hg; fasting blood suger (FBS) as (8.25±3.43,9.02±3.72,9.22±4.62)mmol/L; glycated hemoglobin(HbAlc) as (8.88±2.10,9.34 ± 2.36,9.10 ± 2.36)％ ; uric acid (UA) as (288.04 ± 90.41,307.23 ± 96.96,374.28 ±105.47) mmol/L; triglyceride as (1.72 ± 1.51,2.06 ± 1.88,1.94 ± 1.42) mmol/L,high density lipoprotein cholesterol as ( 1.08 ± 0.30,1.02 ± 0.29,1.07 ± 0.28) mmol/L; fasting insulin as (9.24 ±9.02,11.24 ± 9.74,11.06 ± 9.29) μU/ml; fasting C peptide as (462.31 ± 289.94,510.02 ± 350.08,595.93 ± 445.86) pmol/L.There were significant differences between NA,MA and LA groups in all above items ( P ＜ 0.01 or P＜ 0.05 ).( 3 ) Logistic regression analysis showed that DN were related with duration of diabetes,BMI,SBP,HbAlc,FBS,UA (OR values were 1.041,1.055,1.028,1.116,1.100,1.004 respectively,P＜0.05 or P＜0.01 ).Conclusion It would be helpful to prevent and retard progression of DN that comprehensively controlling high blood glucose,hypertension,hyperuricemia and body weight of type 2 diabetic patients.

OBJECTIVETo study the effect of superparamgnetic iron oxides (ferumoxides) on the survival and proliferation of neural stem cells (NSCs) and determine the optimal ferumoxides concentration for labeling.

METHODSBone marrow stromal cells (BMSCs) were obtained from rat femoral marrow and cultured in vitro to induce their differentiation into NSCs. Ferumoxides labeling of the NSCs was performed with different final concentrations of ferumoxides, and the labeling efficiency and viability of the labeled NSCs were evaluated by Prussian blue staining, MTT assay, flow cytometry and transmission electron microscope.

RESULTSThe NSCs could be effectively labeled with ferumoxides with a labeling efficiency of around 90%. Prussian blue staining showed numerous fine granules with blue staining in the cytoplasm of the labeled NSCs, and the intensity of the blue staining was in positive correlation with the ferumoxide concentration for labeling. Transmission electron microscopy of the labeled NSCs revealed the presence of numerous vesicles spreading in the cytoplasm and filled with electron-dense magnetic iron particles. The ferumoxides vesicles increased with the labeling concentration of ferumoxides, and at the final concentration exceeding 25 microg/ml, ferumoxides vesicles in the NSCs gave rise to conglomeration which hampered observation of the cellular ultrastructure by transmission electron microscope. The results of flow cytometry and MTT assay demonstrated that the cell viability, proliferation, differentiation and apoptosis of the labeled cells were affected by ferumoxides at the concentration above 25 microg/ml, but such effects could be minimal at lower concentrations.

CONCLUSIONFerumoxides might be feasible for in vitro labeling of the NSCs with the optimal concentration of 25 microg/ml.

Animals ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Cells, Cultured ; Dextrans ; Ferrosoferric Oxide ; Iron ; pharmacology ; Magnetite Nanoparticles ; Male ; Microscopy, Electron, Transmission ; Neurons ; cytology ; ultrastructure ; Oxides ; pharmacology ; Rats ; Stem Cells ; cytology ; ultrastructure

AIMTo evaluate the hemodynamic effects of aminophylline and nifedipine in patients with HAPE.

METHODS10 patients with HAPE undergone Swan-Ganz catheter. The parameters of hemodynamics and arterial blood gases in HAPE were measured before and after administration of nifedipine 20 mg sublingually and aminophylline 0.25 g intravenously respectively.

RESULTSAfter administering 0.25 g aminophylline the mPAP and PVR significantly decreased, the cardiac output and the level of PaO2, SaO2 increased obviously, the mSAP, HR did not change so much. After using 20 mg nifedipine, the mPAP, PVR and mSAP also decreased, while the cardiac output, HR and the level of PaO2, SaO2 did not show any changes.

CONCLUSIONBoth of aminophylline and nifedipine can attenuate pulmonary hypertension in patients with HAPE, but the effect of aminophylline was better than the effect of nifedipine.

Adult ; Altitude ; Altitude Sickness ; drug therapy ; Aminophylline ; administration & dosage ; therapeutic use ; Humans ; Hypertension, Pulmonary ; drug therapy ; Male ; Nifedipine ; administration & dosage ; therapeutic use ; Treatment Outcome

BACKGROUNDOxidative stress such as low-density lipoprotein (LDL) oxidation is thought to be an important mechanism in Alzheimer's disease (AD). Paraoxonase 1 (PON1), an enzyme located on high-density lipoprotein, can prevent LDL from oxidation to some extent. It is also a potent cholinesterase inhibitor and an arylesterase, combating organophosphate poisoning and metabolization of environmental neurotoxins which might be responsible for neurodegeneration with aging. We evaluated the association of Gln192Arg polymorphism in the PON1 gene with AD in a Chinese Han ethnic population.

METHODSPatients and age-matched controls were recruited from outpatient clinics and a population-based epidemiological survey, respectively. Gln192Arg polymorphism in the PON1 gene was detected by allele-specific PCR technique in 521 patients with AD and 578 healthy controls.

RESULTSThe presence of at least one of PON1 R alleles (Q/R or R/R) was lower in AD patients than in the controls (82.7% vs 87.4%; chi(2) = 4.68, P = 0.03). PON1 gene R allele frequency was lower in AD patients than in the controls (60.7% vs 64.7%; chi(2) = 3.85, P = 0.05). One-way ANOVA showed that PON1 genotype had no effect on the age of onset for developing AD. Logistic regression analysis demonstrated the age and sex-adjusted odds ratio (OR) for the risk of AD in PON1 of PON1 R allele carriers was 0.71 (P = 0.044, 95% CI, 0.51 - 0.99).

CONCLUSIONOur results indicate that Gln192Arg polymorphism in the PON1 gene is associated with AD, and PON1 R allele might be a protective factor for AD in a Chinese Han ethnic population.

Aged ; Aged, 80 and over ; Alzheimer Disease ; genetics ; Aryldialkylphosphatase ; genetics ; China ; ethnology ; Female ; Genotype ; Humans ; Male ; Middle Aged ; Polymorphism, Single Nucleotide

OBJECTIVETo investigate the feasibility, safety and oncological effect of totally laparoscopic total gastrectomy (TLTG).

METHODSThe clinical data of TLTG cases and open total gastrectomy (OTG) patients between November 2007 and October 2011 were analyzed. Also compared the feasibility, safety and short-term outcomes of TLTG with OTG.

RESULTSNinty cases were analyzed. There were 18 cases in the TLTG group and 72 cases in the OTG group. Operation time was significantly longer in the TLTG group ((310 ± 86) minutes) than in the OTG group ((256 ± 57) min, t = 4.963, P = 0.002), However, the blood loss were significantly lower in the TLTG group ((136 ± 84) ml vs. (359 ± 141) ml, t = -11.734, P = 0.000). The post operative morbidity was similar between the TLTG and OTG group. First flatus time (t = -7.020), first diet time (t = -6.166 and -5.698), and post operative hospital stay (t = -4.610) were significantly shorter in the TLTG group than in the OTG group (P < 0.05).

CONCLUSIONSLTG is a safe and feasible procedure with quick post-operation recovery. The laparoscopic side-to-side esophagojejunal anastomosis is a safe and feasible method of alimentary reconstruction after laparoscopic total gastrectomy.

Adult ; Aged ; Female ; Gastrectomy ; methods ; Humans ; Laparoscopy ; Laparotomy ; Length of Stay ; Lymph Node Excision ; Male ; Middle Aged ; Stomach Neoplasms ; surgery ; Treatment Outcome