Early diagnosis of primary ureteral neoplasms:report of 28 cases / 中国基层医药

Objective To evaluate the choice of early diagnosis method of primary ureteral neoplasms in or-der to improve the ratio of clinical diagnosis. Methods 28 cases with primary ureteral neoplasms were retrospectively analyzed. Ultrasonic examination, IVU, retrograde urogram, spiral CT, MRI, ureteroscopy and exfoliative cell examina-tion of urine were compared in this study. Results The most useful methods of detecting tumors preoperation were retrograde urogram, spiral CT, MRI, ureteroseopy. All the 28 patients underwent surgical treatment. Among them, nephroureterectomy and bladder cuff or partial resection were performed in 19 cases. Postoperative pathology showed transitional cell carcinoma in 27 cases,and adenoma in 1 case. 8 cases were T1-2 tumours. Of the 14 cases during 1990 ～1999 period, 1,5,3,2,2 and 1 cases had survival time of 1,2,3,4,5 and 6 years ,respectively. Of the 14 cases during 2000～2007,4 were lost to follow-up;2 survived for 3 years and 2 for 1 year;the other 6 who have survived near 5 years have been followed till now. Conclusions To improve the early diagnosis rate,B-ultrasonic examination, IVU,retrograde urogram,3D spiral CT and MRI examination were necessary in the early stage. The patients should be opeiated as early as possible after diagnosis.

Determination of binding ability of PaP3’s terminase large subunit to cos site / 第三军医大学学报

Objective To investigate the binding ability of the terminase large subunit of Pseudomonas aeruginosa bacteriophage PaP3 to the cos site. Methods The gene tls was amplified from the genome of bacteriophage PaP3 by PCR and subcloned into pMD18-T vector. Then the gene tls cut down from the vector was inserted into the plasmid pQE31 which could give a 6-His tag at the N-terminal of the expressed protein. The recombinant vector pQE-tls was transformed to E.coli. JM109, after induction with IPTG, the expressed bacteria were resuspended and sonicated, then after centrifugation, the inclusion body was obtained. The inclusion body was dissolved with lysis buffer, then the tagged protein was purified by Ni-NTA affinity chromatography and renatured by dialysis. Finally the DNA-binding ability of the fusion protein rTLS was determined by EMSA. Results The expression plasmid pQE31-tls was successfully constructed, and the target protein yield was up to 30% of the total bacterial proteins. After purification and renaturation, the fusion protein rTLS can partially bind the cos fragment. Conclusion The fusion protein rTLS was successfully expressed, purified and renatured. The rTLS has the specific DNA-binding activity. The present work lays the foundation for the further research of the gene tls.

Cloning of LVA Ca 2＋ channel and its primary structure features / 细胞与分子免疫学杂志

Aim To clone LVA calcium channel (also known as T-type calcium channel) from INS-1 cell line derived from rat pancreatic β cells. Methods RT-PCR, 3′ -RACE and 5′ -RACE were used to clone coding sequence of the whole gene. DNA sequencing and genomic DNA walking were used to identify the primary structure features and exons. Results The cloned gene, named as α 1 G-INS, was composed of 6 864 bp, encoding 2 288 amino acids, which shares 96.3％ identity to α 1G, the neuronal T-type calcium channel. Compared to α 1G, the amino acids in membrane-spanning regions of four transmembrane domains and intracellular loop LI-II of α 1G-INS were highly conserved, but there are three distinct regions. This discrepancy was due to alternative mRNA splicing. Scattering on other regions of the molecule there were 10 single amino acid substitutions, which could be explained as site-mutations of the gene. Conclusion A new isoform,α 1G-INS, of T-type calcium channel is successfully cloned from rat insulin-secreting cell line INS-1, which is significant to further understanding many Ca2＋ -involved biologically essential processes.