Objective To investigate the correlation rs2272087 polymorphism of STATS gene and asthma.Methods The polymerase chain reaction PCR- SBT technique was used to determine rs2272087 polymorphism in asthma and control group.Results The genotype of AA,AG and GG of rs2272087 were 0.600,0.412 and 0.167 in asthma group,and 0.430,0.200,0.367 in control group,respectively.The frequency of allele A and G was 0.903 and 0.344 in asthma group,and 0.656,0.970 in control group,respectively.There was significant difference in two groups(x2 =9.40,P ＜0.01 ;x2 =11.58,P ＜0.01 ).Conclusions The rs2272087 polymorphism of STAT5 gene may be an important candidate gene for asthma.

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.

OBJECTIVEEpidemiologic study on the perinatal interaction of antioxidant vitamins and heavy metals on offspring's intellectual development is extremely limited. The aim of this prospective cohort study was to investigate the status of maternal and cord blood antioxidant vitamins (Vitamin A, E and C) and heavy metals (lead, cadmium and mercury) at delivery and correlations between these antioxidant vitamins or metals and neurodevelopment of early childhood, and to explore the protective effect of antioxidant vitamins against the injure of heavy metals to intellectual development in children in Tongliang, Chongqing, China.

METHODA total of 150 pairs of mothers-neonates were recruited. Serum concentrations of vitamin(V) A, vitamin E, vitamin C, lead, cadmium and mercury in maternal and cord blood after delivery were determined and intellectual development was evaluated by Gesell Development Schedule (GDS) at two years of age.

RESULTA total of 111 pairs of maternal-neonatal subjects with completed data were included into statistical analysis. After adjusting for potential confounders, cord blood mercury level was found negatively associated with all the development quotients (DQs) (β = -0.486, β = -0.716, β = -0.846, β = -0.935, β = -0.702, P < 0.05). Cord VE level was positively correlated with motor, adaptation, language and average DQ (β = 0.475, β = 0.458, β = 0.403, β = 0.395, P < 0.05). When antioxidant vitamins and heavy metals levels were all included into the multiple regression model, cord vitamin E level was statistically positively related with motor, adaptation and average DQs (β = 0.449, β = 0.412, β = 0.349, P < 0.05). While cord blood mercury level was negatively correlated with adaptation, language, social and average DQ (β = -0.397, β = -0.727, β = -0.935, β = -0.628, P < 0.05), the relationship between cord mercury level and motor DQ was not statistically significant (P = 0.0890). The motor, adaptation, language and average DQs in high cord VE group were higher than those in low VE group (t = 2.93, t = 2.06, t = 2.13, t = 2.27, P < 0.05). Social DQ in high cord lead group was significantly lower than that in low lead group (χ(2) = 5.56, P = 0.015). Cord VC level in high cadmium group was significantly lower than that in low cadmium group (χ(2) = 7.62, P = 0.006). VA placental transport ratio in high mercury group was significantly lower than those in low mercury group (χ(2) = 8.02, P = 0.005).

CONCLUSIONOur data suggested that there may be certain interaction between antioxidant vitamin A, E, C and heavy metals lead, cadmium, mercury. Antioxidant vitamins at delivery may have protective effect on the early neurodevelopment via interaction with the heavy metals.

Adult ; Antioxidants ; analysis ; Child Development ; Child, Preschool ; Cohort Studies ; Female ; Humans ; Infant, Newborn ; Intelligence Tests ; Metals, Heavy ; blood ; Pregnancy ; Prospective Studies ; Surveys and Questionnaires ; Vitamins ; blood

Brain magnetic resonance imaging (MRI) of the elderly often reveals white matter changes (WMCs) with substantial variability across individuals.Our study was designed to explore MRI features and site-specific factors of ischemic WMCs.Clinical data of consecutive patients diagnosed with ischemic cerebral vascular disease who had undergone brain MRI were collected and analyzed.Multi-logistic regression analysis comparing patients with mild versus severe WMCs was performed to detect independent associations.Analyses of variance (ANOVAs) were used to detect regionally specific differences in lesions.We found that lesion distribution differed significantly across five cerebral areas,with lesions being predominant in the frontal lobe and parieto-occipital area.To explore WMCs risk factors,after adjusting for gender,diabetes mellitus,and hypertension,only age (P＜0.01),creatinine (P=0.01),alkaline phosphatase (ALP) (P=0.01) and low-density lipoprotein cholesterol (LDL-C) (P=0.03) were found to be independently associated with severe WMCs.Age (P＜0.001) was strongly associated with WMCs in the frontal lobe while hypertension was independently related to lesions in the basal ganglia (P=0.048) or infratentorial area (P=0.016).In conclusion,MRI of WMCs showed that ischemic WMCs occurred mostly in the frontal lobe and parieto-occipital area.The infratentorial area was least affected by WMCs.Typically,age-related WMCs were observed in the frontal lobes,while hypertension-related WMCs tended to occur in the basal ganglia and infratentorial area.

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

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

OBJECTIVETo explore the etiopathogenesis, therapy and incidence of pulmonary infection in kidney transplantation recipients taking new immunosuppressant.

METHODSThe clinical data from 752 kidney transplant recipients were retrospectively analyzed, who were divided into 3 groups according to the immunosuppressants administered, namely group A (CsA+MMF+Pred, n=226), group B (FK506+MMF+Pred, n=386) and group C (FK506+Rap+Pred, n=140). The incidence and mortality of pulmonary infection were recorded and the analysis of etiopathogenesis, diagnosis and therapy of pulmonary infection were carried out in the 3 groups.

RESULTSFifty-three patients acquired post-transplant pulmonary infection. The incidence of pulmonary infection was 7.08% (16/226) in group A, 7.25% (28/386) in group B and 6.43% (9/140) in group C. One patient died in group A and 2 in group B. Among the 53 patients, 24 had simple bacterial infection, 9 had cytomegalovirus infection, 1 had mycotic infection, 17 had combined infection, and 2 had unidentified pathogen infection. Of the pathogenic bacteria detected, 68.35% were Gram-negative.

CONCLUSIONGram-negative bacteria are most likely responsible for pulmonary infection after kidney transplantation, which most possibly occurs within 6 months after kidney transplantation. Early diagnosis and early treatment are critical for decreasing the mortality of severe pneumonia and for improving the survival rate of the patients and grafts.

Adolescent ; Adult ; Aged ; Cyclosporine ; administration & dosage ; adverse effects ; Cytomegalovirus Infections ; chemically induced ; diagnosis ; therapy ; Female ; Gram-Positive Bacterial Infections ; chemically induced ; diagnosis ; therapy ; Humans ; Immunosuppressive Agents ; administration & dosage ; adverse effects ; Kidney Transplantation ; Lung Diseases ; chemically induced ; diagnosis ; therapy ; Male ; Middle Aged ; Tacrolimus ; administration & dosage ; adverse effects

OBJECTIVE: To establish the 2-dimensional electrophoresis(2-DE) profiles of peripheral blood mononuclear cells(PBMC) in patients with hepatocellular carcinoma(HCC) and health adults. METHODS: The total proteins from PBMC in patients with HCC and healthy adult were separated by immobilized pH gradient-based 2-DE. The differential expression proteins were analyzed by PDQuest analysis software. RESULTS: The well-resolved, reproducible 2-DE patterns of PBMC in patients with HCC and healthy adults were obtained. For HCC, the average spots of 2-DE maps were 1 206 +/- 48, and the average matching rate was 90.8%. For normal adults, the average spots were 1 123 +/- 37, and the average matching rate was 92.6%. CONCLUSION The well-resolved, reproducible 2-DE patterns of PBMC in patients with HCC and healthy adults are established. These proteomic analysis methods are useful to screen the potential biomarkers in the early diagnosis, treatment and prognosis monitor in patients with malignant tumor.
Adult ; Carcinoma, Hepatocellular ; blood ; Electrophoresis, Gel, Two-Dimensional ; methods ; Female ; Humans ; Leukocytes, Mononuclear ; cytology ; metabolism ; Liver Neoplasms ; blood ; Male ; Middle Aged ; Proteomics ; methods

BACKGROUNDCervical keratinocytes are recovered at a low numbers and frequently associated with contaminating human fibroblasts which rapidly overgrow the epithelial cells in culture with medium supplemented with 10% fetal bovine serum (FBS). However, it is difficult to initiate keratinocyte cultures with serum-free keratinocyte growth medium alone because cell attachment can be poor. Therefore, the culture of these cells is extremely difficult. In this study, we described a modified culture medium and coated culture plastics for growing normal human cervical epithelial cells in vitro.

METHODSNormal cervical epithelial tissue pieces were obtained and digested with type I collagenase to dissociate the cells and a single cell suspension produced. The cells were cultured on plastic tissue culture substrate alone or substrate coated with collagen type I from rat tail, with modified keratinocyte serum-free medium (K-SFM) supplemented with 5% FBS. After attachment, the medium were replaced with K-SFM without FBS. The expression of basal keratins of the ectocervical epithelium, K5, K14 and K19 were assayed by immunofluorescence with monoclonal antibodies to identify the cell purity.

RESULTSOur results indicate that cells attached to the culture plastic more quickly in K-SFM supplemented with 5% FBS than in K-SFM alone, as well as to tissue culture plastic coated with collagen type I than plastic alone. The modified medium composed of K-SFM and 5% FBS combined with a specific tissue culture plastic coated with collagen type I from rat tail was the best method for culture of normal cervical epithelial cells. K5, K14 and K19 were assayed and keratinocyte purity was nearly 100%.

CONCLUSIONA novel, simple and effective method can be used to rapidly obtain highly purified keratinocytes from normal human cervical epithelium.

Cell Culture Techniques ; methods ; Cervix Uteri ; cytology ; Epithelial Cells ; cytology ; Female ; Humans ; Keratinocytes ; cytology