Objective To assess quickly the potential environmental pollution by using the luciferase transgenic cells. Methods Many electrophile compounds in the environment can generate oxidative stress,so that the transcription of certain protective genes is induced via specific DNA motifs called electrophile response elements (EPREs). We have made a vector containing a single EPRE fused to the TK minimal promoter and the gene encoding firefly luciferase (EPRE-LUC) by adopting bio-molecular techniques. From this vector the stable LUC expression vector regulated by EPRE had been successfully reconstructed. This reporter construct was transfected into HeLa cells,and the clones resistant to G418 were selected. The resistant cells were treated by the different concentrations of sodium arsenate(NaAsO2),cadmium chloride (CdCl2),mercury chloride(HgCl2) and diethyl mateate (DEM). After that,the expression of luciferase was determined by luciferase assay kit. Results The correct construct frame of LUC reporter vector regulated by EPRE was identified by DNA sequencing; the dose-dependent relationships between the LUC expression and the test substance concentration were found. Among them,the relationship produced by DEM was the most significant. Conclusion The LUC transgenic cells regulated by EPRE have been successfully constructed.

Objective To develop a rapid screening method of environmental estrogen in vitro. Methods Estrogen responsive element (ERE) was inserted into the upstream of Lac Z (?-galactosidase ) reporter gene and the reporter gene was regulated under estrogen receptor(ER) in the recombinant yeast cells by the molecular biological technique. When the yeast cells was affected by environmental estrogen, conformational change of ER lead to expression through binding to ERE. There exist the functional relationships between the expression and the pollution degree of environmental estrogen. Therefore, the pollution degree can be reflected by assaying the activity of ?-galactosidase. Results It was identified by PCR that recombinant yeast cell, whose Lac Z reporter gene regulated under estrogen receptor, was successfully reconstructed. The experimental results showed that the time-dose-effect response had no significant changes when the recombinant yeast cell bad been exposed to ?-estradiol for 4 and 8 hours respectively; among the three methods of yeast disintegration, the ultrasonic vibration method had more advantage compared with that of the other methods and this dose-effect curve exhibited "S" shape; among 3 test chemical products, ?-estradiol was the most estrogenic activity, biosphenol A take second place, estradiol benzoate was the lowest. Conclusion The assay system of yeast cell for environmental estrogen hormone mediated by estrogen receptor is stable and applicable.

Objective Ankylosing spondylitis (AS) can affect both the lumbar zygapophyseal joint and the centrum .This study was to compare multislice spiral CT ( MSCT) and MRI in the diagnosis of zygapophyseal joint lesions in AS patients and assess the role of zygapophyseal joint lesions in the early diagnosis of AS . Methods We retrospectively analyzed the lumbar imaging data of 41 male patients with AS .Forty-one male AS patients underwent MSCT , 18 receiving normal MRI , and the other 23 diffusion weighted imaging (DWI) and CE-T1WI-STIR in addition.Using Fisher′s Exact Test, we compared MSCT and MRI in their detection rates of a-pophyseal joint lesions and positive changes in the zygapophyseal joint and lumbar centrum .Then we analyzed the relation between the zygapophyseal joint lesions and the disease duration . Results The detection rates of zygapophyseal joint and centrum lesions were 90.2%and 58.5%on MSCT (P>0.05), and 80.5%and 46.3%on MRI (P>0.05), respectively.MSCT and MRI exhibited sig-nificant differences in the detection rate of centrum lesions (P<0.05) but not in that of zygapophyseal joint lesions (P>0.05). These lesions could appear within 1 year after the onset of AS or ahead of vertebral changes . Conclusion Both MSCT and MRI can manifest zygapophyseal joint lesions , which may develop in the lumbar spine at the early stage of AS , ahead of centrum lesions .This is important for the early diagnosis of AS .

OBJECTIVETo evaluate the diagnostic value of pelvis bone marrow fat depositions (BMFD) displayed by magnetic resonance imaging (MRI) in patients with ankylosing spondylitis (AS).

METHODSEighty-eight subjects undergoing pelvic MRI examinations were enrolled in this study, including 44 with clinically confirmed AS (39 male and 5 female patients with a mean age of 26.41∓8.09 years) and 44 control subjects without AS (37 male and 7 female subjects with a mean age of 29.32∓7.31 years). The incidence of BMFD in the bilateral sacroiliac (SI) joints and acetabulum were compared between the two groups. The distribution features of BMFD of the periarticular cancellous bone marrow in the pelvis and in other regions of the pelvis were analyzed for the AS patients, and the incidence of BMFD was determined in different stages of sacroiliitis and hip arthritis.

RESULTSThe incidence of BMFD in the SI joints and acetabulum was significantly higher in the AS patients than in the control subjects (P<0.01); The incidence of BMFD was significantly higher in the periarticular cancellous bone marrow than in the other positions of pelvis (P<0.01). The incidence of BMFD ranged from 40.0% to 45.9% in early stages of sacroiliitis, significantly lower than the incidence in later stages (58.3%-73.1%, P<0.01); the incidence showed no difference between different stages of hip arthritis (P>0.01).

CONCLUSIONSAS patients have a higher incidence of BMFD in the pelvis than control subjects. BMFD is distributed mainly under the articular surface, seen throughout the stages of AS, indicating that BMFD is an important pathological change of the bone marrow in AS to potentially allow early diagnosis of AS.

Adipose Tissue ; pathology ; Adolescent ; Adult ; Bone Marrow ; pathology ; Case-Control Studies ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Pelvis ; pathology ; Spondylitis, Ankylosing ; pathology ; Young Adult

When we try to establish the gene recombinant yeast cell to screen the androgenic endocrine disruptors, the key procedure is the androgen receptor (AR) expression in the yeast cell. For this purpose, we obtained the GPD (glyceraldehyde-3-phosphote dehydrogenase) promoter from the yeast genosome of W303-1A using PCR system and inserting it into Swa I and BamH I sites of pYestrp2. The new constructed vector was named pGPD. The V5 epitope tag DNA with a 5'-BamH I and a 3'-EcoR I sticky end was cloned into the corresponding site of the pGPD vector to yield the vector of pGPDV5. The 2 723 bp full length AR ORF amplified by PCR from pcDNA3.1/AR was fused to V5 epitope tag DNA in pGPDV5 to give the AR yeast expression vector of pGPDV5/AR. This fused vector was transformed into the yeast cell (W303-1A). Western blot was used to detect the V5 fused protein of AR, in the protocol of which the primary monoclonal antibody (IgG(2a)) of mouse anti-V5 and the polyclonal secondary antibody of goat anti-mouse (IgG) linked to horseradish peroxidase (HRP) were used to detect the specific protein in the given sample of the transformed yeast extract. The result showed that the fused protein of AR was expressed successfully in the yeast cell.
Base Sequence ; Endocrine Disruptors ; analysis ; Epitopes ; biosynthesis ; genetics ; Genetic Vectors ; genetics ; Glyceraldehyde-3-Phosphate Dehydrogenases ; genetics ; Humans ; Molecular Sequence Data ; Promoter Regions, Genetic ; Receptors, Androgen ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; genetics ; Yeasts ; genetics ; metabolism