Objective To investigate the value of high-dose dobutamine stress echocardiography combined with two-dimensional strain imaging in early diagnosis of coronary artery disease. Methods Highdose dobutamine stress echocardiography was performed to 28 patients with suspected coronary artery disease. All wall movements were observed during resting condition and at all stress levels,respectively;the peak systolic longitudinal strain in each endomyocardial segment of left ventricular was measured; the sensitivity and specificity between visual method and two-dimensional strain imaging in diagnosing myocardial ischemia with high-dose dobutamine stress echocardiography were compared. The average peak systolic longitudinal strain was calculated against control group, coronary artery disease group during ischemia segments and non-ischemia segments, and a comparison was made inside each group as well as against the other groups. The area under receiver operating characteristic curve of the peak systolic longitudinal strain was used to predict the sensitivity and the specificity of myocardial ischemia. Results With dobutamine dose of 40 μg·kg-1 · min-1 ,wall motion abnormalities were diagnosed in 6 patients (20 segments) through visual method, myocardial ischemia was found in 15 patients (148 segments) through computing the peak systolic longitudinal strain. Inside the coronary artery disease group during ischemic　segments,the majority of peak systolic longitudinal strain was significantly reduced ( P＜0.05) compared to the non-ischemic segments and the control group. In diagnosing myocardial ischemia in high-dose dobutamine stress echocardiography, the sensitivity of visual method and two-dimensional strain imaging were 35.3％ and 88.2％(P＜0.01), specificity 100％ and 100％(P＞0.05), and accuracy 60.7％ and 92.8％ (P＜0.01). The cutoff value of the peak systolic longitudinal strain was less than or equal to 14.9％, its sensitivity and specificity in predicting myocardial ischemia were 83.3％ and 91.7％,respectively. Conclusions High-dose dobutamine stress echocardiography combined with two-dimensional strain imaging can increase the sensitivity of detecting myocardial ischemia and detect concealed myocardial ischemia. High-dose dobutamine stress echocardiography combined with two-dimensional strain imaging can be used in early diagnosis of coronary artery disease.

Objective Aiming at detecting Staphylococcus aureus、Pseudomonas aeruginosa and Klebsiella pneumoniae in laboratory animals,the paper provides a rapid,sensitive and simple test method.Methods According to Staphylococcus aureus nuc gene,Pseudomonas aeruginosa LasI gene,Klebsiella pneumonia PhoE gene and general 16S rRNA gene, designed specific primers;Through the optimization of multiplex PCR primer concentrations and annealing temperature, the specificity and sensitivity of detection, establishing multiplex PCR system.Application of the PCR system test specimens of artificial infections and experiment animal feces is compared with traditional test method.Results Multiplex PCR amplification of Staphylococcus aureus (153 bp), Pseudomonas aeruginosa (600 bp) with Klebsiella pneumoniae (368 bp) and general (520 bp).The multiplex sensitivity for the purpose of 10pg, specificity of detection was not detected from other pathogens.Application of establishing multiplex PCR system to detect the artificial positive samples, and detect 1 Pseudomonas aeruginosa positive case in 76 fecals.Conclusions This paper established the multiplex PCR method which has the advantages of specific,sensitive,simple and rapid, and provides a reliable way for rapid test in laboratory animals microbiology.

Gastrointestinal Hemorrhage ; etiology ; Humans ; Infant, Newborn ; Intestines ; abnormalities ; pathology ; Male ; Necrosis

OBJECTIVETo explore the prevalence, distribution and clinical significance of high-intensity zone (HIZ) in anterior annulus fibrosus (AF) in comparison with HIZ in posterior AF of lumbar disc.

METHODSAccording to the diagnosis and location of HIZ, 610 lumbar magnetic resonance images with entire clinical materials were divided into control group (without HIZ), anterior AF group (HIZ), posterior AF group (HIZ) and anterior & posterior (AP) AF group (HIZ). The incidence of HIZ was summarized. The clinical data, such as male/female ratio, age, and body weight, prevalence of low back pain (LBP) and distribution of HIZ, were compared and analyzed between the groups.

RESULTSThree hundred and fifteen cases shown no HIZ (51.6%), 95 cases presented HIZ in anterior AF (15.6%, 119 discs), 159 cases presented HIZ in posterior AF (26.1%, 189 discs) and 41 cases presented HIZs in both anterior and posterior AF (6.7%, 96 discs). There was significant difference between the prevalence of HIZ in anterior AF and that in posterior AF (P < 0.01). The male/female ratio and body weight of each groups showed no difference (P > 0.05), and the age was proved to be statistically different between four groups (P < 0.01, control group < posterior AF group < AP AF group < anterior AF group). HIZs in anterior AF often occurred at L(1,2)-L(4,5), whereas, they usually developed at L(3,4)-L(5)S(1) in posterior AF. The incidence of LBP in control group, anterior AF group, posterior AF group, AP AF group were 40.0%, 52.6%, 55.4% and 65.8%, respectively. The LBP prevalence of control group was lower than that of other three groups (P < 0.05), and the prevalence of last three groups showed no difference (P > 0.05).

CONCLUSIONSCompared with HIZ in posterior AF, the HIZ in anterior AF of lumbar disc has a lower prevalence, often develops in elder patients and in upper motion segments. It also indicates an obvious relation to LBP as the former.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Intervertebral Disc ; pathology ; Intervertebral Disc Displacement ; diagnosis ; pathology ; Lumbar Vertebrae ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Young Adult

P300/CBP-associated factor(PCAF),an important member of histone acetyltransferase family(HATs) within eukaryotic cells,is capable of inducing the acetylation of histone,promoting the transcription of specific genes and involving in many biological effects.In the present study,full-length cDNA of PCAF was inserted into plasmid pGEX-5x-1,then the soluble protein GST-PCAF was expressed in E.coli BL21(DE3) after the optimization of inducing conditions.The recombinant protein was further purified with affinity chromatography and tested the activity by in vitro acetylation assays.High efficient PCAF protein produced by this method could serve for the study on the role of PCAF in gene regulation and the interaction between PCAF and other proteins.

Adult ; Bone Cysts, Aneurysmal ; complications ; diagnostic imaging ; pathology ; Bone Neoplasms ; etiology ; pathology ; Cell Transformation, Neoplastic ; pathology ; Humans ; Male ; Radiography

OBJECTIVETo observe the changes in the activity of dendritic cells (DCs) after carcino-embryonic antigen (CEA) gene transfection mediated by recombinant adeno-associated virus type2 (rAAV) and tumor cell lysate.

METHODSImmature DCs isolated from peripheral blood monocytes of HLA-A11-positive healthy volunteers were infected with the rAAV carrying CEA gene or loaded with tumor cell lysate. The surface markers of the DCs such as CD40, CD 1alpha, and CD86 were analyzed by flow cytometry. Interleukin-12 (IL-12) in the supernatants of DCs and interferon-gamma (IFN-gamma) released by the cytotoxic T lymphocytes (CTLs) were determined by ELISA detection kit. The specific killing activity of CTL against LoVo cells was assessed by MTT assay.

RESULTSThe DCs following antigen loading with the two methods both highly expressed CD40, CD86 and IL-12, and induced specific CTL that specifically recognized and killed LoVo cells, but the killing effect resulting from rAAV infection of the DCs was much better than that induced by tumor cell lysate loading.

CONCLUSIONBoth methods of antigen loading can induce mature DCs from peripheral blood monocyte cells, but rAAV infection of the DCs can be more effective than tumor cells lysate loading. DCs infected with rAAV may have the potential to serve as an adjuvant immunotherapy for patients with colorectal carcinoma.

B7-2 Antigen ; metabolism ; CD40 Antigens ; metabolism ; Cancer Vaccines ; biosynthesis ; immunology ; Carcinoembryonic Antigen ; genetics ; Cell Line, Tumor ; Colorectal Neoplasms ; therapy ; Dendritic Cells ; immunology ; metabolism ; Dependovirus ; genetics ; Genetic Vectors ; Humans ; Interleukin-12 ; metabolism ; Transfection

OBJECTIVETo construct a mouse that specifically expresses Cre recombinase in hepatocyte.

METHODSA hepatocyte specific transgenic construct containing mouse albumin promoter, the Cre recombinase gene and the poly (A) of human growth factor gene was generated. The linearized constructs were introduced into the fertilized eggs by microinjection to obtain the transgenic mice. The transcriptional specificity of Cre recombinase was detected by reverse transcription polymerase chain reaction (RT-PCR). The expression and function of Cre recombinase were detected by PCR and Southern Blot after crossing the Alb-Cre transgenic mice with the Smad4 conditional knockout mice.

RESULTSThe linearized constructs were microinjected into 837 fertilized eggs, and then the 797 effective eggs of microinjected eggs were implanted into the oviducts of 27 pseudo pregnant mice. In the 53 offspring, there were 6 mice carrying the transgene identified by polymerase chain reaction (PCR) and Southern Blot. Cre recombinase transcripts were detected in the livers and testis of the Alb-Cre transgenic mice using RT-PCR. The Cre recombinase was expressed in the livers of the double heterozygous for Alb-Cre and Smad4 floxed allele, and the exon 8 floxed by loxP site was deleted.

CONCLUSIONA hepatocyte-specific Cre transgenic mouse was generated successfully. The Cre recombinase expressed specifically in liver and could mediate the recombination between loxP sites in vivo.

Animals ; Female ; Hepatocytes ; metabolism ; Humans ; Integrases ; genetics ; Mice ; Mice, Transgenic ; Viral Proteins ; genetics

A method was evaluated for determination of twenty-one doping (including nandrolone, boldenone and methandienone) in human urine by gas chromatography-high resolution mass spectrometry. Samples were prepared by liquid-liquid extraction, concentrated, TMS derivatization and limit of detection at ng x mL(-1) by MID/GC/HRMS. According to the code of the World Anti-Doping Agency (WADA), precision and recoveries of the procedure were evaluated by replicate analysis (n = 6), the recoveries in the range of 66%-103%, with the RSD below 10.0%. The precision within the day of the method with three different concentrations was also determined RSD were less than 9.5%, 10.0% and 9.7%.
Anabolic Agents ; urine ; Doping in Sports ; Gas Chromatography-Mass Spectrometry ; Humans ; Methandrostenolone ; urine ; Nandrolone ; urine ; Substance Abuse Detection ; methods ; Testosterone ; analogs & derivatives ; urine

In the present study, isoelectronic focusing with different pH gradients (pH 3-5, 2-6) or migrating distances (8.5, 12 and 17 cm) and SDS-PAGE was used to separate continuous erythropoietin receptor activator (CERA), recombinant human erythropoietin (rhEPO), darbepoetin and endogenous EPO spiked in human urine with 37 degrees C overnight incubation. Double blotting and chemiluminescent visualization were used to detect the IEF and SDS-PAGE profiles. The bands of CERA profile were detected and well separated from the endogenous EPO and the other two EPO preparations with both SDS-PAGE and the IEF method using a gradient pH 3-5 and a migrating distance of 17 cm, and a significant particular band of CERA profile was found in the IEF result. These preliminary results indicated that the methods were reliable and reproducible for detecting CERA, and could be used as a routine procedure for anti-doping analysis.
Electrophoresis, Polyacrylamide Gel ; Erythropoietin ; urine ; Humans ; Isoelectric Focusing ; methods ; Polyethylene Glycols ; Recombinant Proteins