Objective To analyze the conding region of hantanvirus S gene and predict the structure of nucleoprotein for diagnostic antigen study.Methods RT-PCR was used to amplify the S gene of hantanvirus Hunan03 strain after designing specific primers.The amplification product was cloned into pGM-T vector and then the recombinant vector was transformed into E.coli TOP10,gene sequencing was carried out after blue-white selection and PCR screening for positive clones.The database of NCBI and Swiss-Prot/TrEMBL were used to predict and analyze the structure,biological characteristics and protein structures of S gene.Results The amplification product was about 1290 bp,the pGM-T/S vector was constructed and successfully sequenced,the whole length of the open reading frame (ORF) was composed of 1290 nucleotide residues,among them the GC content was 44.11％ and the AT content was 55.89％,it was composed of 429 amino acids (20 kinds),the accession number of the sequence submitted to GenBank was JN712306,its homology of nucleotides to the 76-118 strain was 83％ and the homology of amino acids was 98％,ten nonspecific variation sites were found.The grand average of hydropathicity was-0.405.There were three transmembrane domains and four non transmembrane domains in the secondary structure of nucleoprotein including 55％ of helix structure,6.1％ of sheet structure and 38.9％ of loop structure.Conclusion The bioinformatics analysis of Hunan03 strain S gene might be important for provide the substructure data to reveal the significance of S gene characteristics on hemorrhagic fever renal syndrome (HFRS) prevention and control.

Objective To establish a new canine model of ureter trauma to observe the protective effect of biodegradable ureteral stent on renal function following traumatic ureter injury. Methods A self-made device was used to make firearm fragment wounds unilaterally on the ureters in nine Beagle dogs (model group). The wounds were debrided and sutured and the results were evaluated by using intravenous pyelography (IVP) and radioactive renography at 40, 80 and 120 days postoperatively. Firearm fragment wounds were made to the bilateral ureters of nine Beagle dogs in the positive control group, in which a biodegradable stent was placed in one side and a double-J stent placed in the other side. Results In model group, hydronephrosis and hydroureter occurred and got worse postoperatively on the wounded side in all nine Beagle dogs, while none of these symptoms were found in any animals in the control group. The ratios of biodegradable stent side to double-J stent side were increased in renal patial concertration index and half time of kidney washout, but neither showed significant differences. However,vesicoureteral reflux (VUR) was not found in the biodegradable stent side, but in the double -J stent side. Conclusions A new canine model of firearm fragment wounds is successfully developed. Both the biodegradable and double-J stent play important roles in support and drainage and show no significant difference in aspects of renal uptake and half time of kidney washout. The biodegradable stent can effectively prevent VUR.

Histone acetylation is a well-characterized epigenetic modification controlled by histone acetyltransferases (HATs) and histone deacetylases (HDACs). Imbalanced histone acetylation has been observed in many primary cancers. Therefore, efforts have been made to find drugs or small molecules such as HDAC inhibitors that can revert acetylation levels to normal in cancer cells. We observed dose-dependent reduction in the endogenous and exogenous protein expression levels of KAT8 (also known as human MOF), a member of the MYST family of HATs, and its corresponding histone acetylation at H4K5, H4K8, and H4K16 in chemotherapy drug gemcitabine (GEM)-exposed T24 bladder cancer (BLCA) cells. Interestingly, the reduction in MOF and histone H4 acetylation was inversely proportional to GEM-induced γH2AX, an indicator of chemotherapy drug effectiveness. Furthermore, pGL4-MOF-Luc reporter activities were significantly inhibited by GEM, thereby suggesting that GEM utilizes an MOF-mediated anti-BLCA mechanism of action. In the CCK-8, wound healing assays and Transwell ® experiments, the additive effects on cell proliferation and migration were observed in the presence of exogenous MOF and GEM. In addition, the promoted cell sensitivity to GEM by exogenous MOF in BLCA cells was confirmed using an Annexin V-FITC/PI assay. Taken together, our results provide the theoretical basis for elucidating the anti-BLCA mechanism of GEM.

OBJECTIVETo investigate the relationship between the molecular characteristics and phylogenetic evolution of rabies N gene.

METHODSSaliva samples were collected from rabies cases, and RT-PCR was used to amplify the N gene of rabies virus with the specific primers. The amplifying product of RT-PCR was cloned to pUCm-T vector and transformed into E.coli XL1-Blue and then the blue-white selection, PCR screening and gene sequencing were carried out to identify the positive clones. Finally, ExPASy and other bioinformatics software were used to analyze and predict the structure and biological characteristics of the N genome.

RESULTSThe amplification product of RT-PCR was 1 353 bp, the recombinant plasmid pUCm-T/N was constructed, the whole length of the N gene open reading frame was composed of 1 353 nucleotide residues to code 450 amino acids (20 kinds), the accession number submitted to the Genbank was HM756692, its sequence homology of nucleotides and amino acids compared with the vaccine strain CTN-1-V was 90% and 99% respectively. The evolutionary analysis showed that the isolated strain belonged to genotype I with certain geographic regionality.

CONCLUSIONThe characteristics investigation and bioinformatics analysis of Hunan0806 N gene will provide fundamental data to reveal the significance of the N gene characteristics for rabies epidemiology and its prevention & control.

Amino Acid Sequence ; Gene Expression Regulation, Viral ; physiology ; Humans ; Models, Molecular ; Molecular Sequence Data ; Nucleocapsid Proteins ; genetics ; metabolism ; Phylogeny ; Protein Conformation ; Rabies ; virology ; Rabies virus ; genetics ; metabolism ; Saliva ; virology

Objective　: To investigate the protective effects of different types of lead collars on the thyroid during radio- therapy after breast-conserving surgery.  Methods: Forty breast cancer patients undergoing radiotherapy after breast-con- serving surgery were randomly divided into four groups to wear different lead collars for thyroid protection: control group (0 mm Pb), common material group (0.5 mm Pb), common material group (2 mm Pb), and new radiation-shielding material group (2 mm Pb). Radiation doses inside and outside lead collars were monitored. A questionnaire survey was conducted to acquire information on patient acceptance of the lead collars. Results: All the groups (except the control group) showed significant differences between scattered radiation doses inside and outside lead collars (P < 0.05). The scattered radiation was attenuated by 33.64% on average in the 2-mm new material group, which was significantly higher than in the other groups (P < 0.05). After radiotherapy, there was no significant change in the color and appearance of skin under lead collars in any group. All the patients were normal at the first thyroid ultrasound re-examination. The 2-mm new material lead collar was the most acceptable. Conclusion　 The lead collar made of the new radiation-shielding material has a good protective effect on the thyroid gland, and is easily accepted by patients, which can be promoted for application.

The BCCIP (BRCA2- and CDKN1A-interacting protein) is an important cofactor for BRCA2 in tumor suppression. Although the low expression of BCCIP is observed in multiple clinically diagnosed primary tumor tissues such as ovarian cancer, renal cell carcinoma and colorectal carcinoma, the mechanism of how BCCIP is regulated in cells is still unclear. The human INO80/YY1 chromatin remodeling complex composed of 15 subunits catalyzes ATP-dependent sliding of nucleosomes along DNA. Here, we first report that BCCIP is a novel target gene of the INO80/YY1 complex by presenting a series of experimental evidence. Gene expression studies combined with siRNA knockdown data locked candidate genes including BCCIP of the INO80/YY1 complex. Silencing or over-expressing the subunits of the INO80/YY1 complex regulates the expression level of BCCIP both in mRNA and proteins in cells. Also, the functions of INO80/YY1 complex in regulating the transactivation of BCCIP were confirmed by luciferase reporter assays. Chromatin immunoprecipitation (ChIP) experiments clarify the enrichment of INO80 and YY1 at +0.17 kb downstream of the BCCIP transcriptional start site. However, this enrichment is significantly inhibited by either knocking down INO80 or YY1, suggesting the existence of both INO80 and YY1 is required for recruiting the INO80/YY1 complex to BCCIP promoter region. Our findings strongly indicate that BCCIP is a potential target gene of the INO80/YY1 complex.
Calcium-Binding Proteins ; genetics ; metabolism ; Cell Cycle Proteins ; genetics ; metabolism ; Chromatin Assembly and Disassembly ; physiology ; DNA Helicases ; genetics ; metabolism ; HeLa Cells ; Humans ; Multiprotein Complexes ; genetics ; metabolism ; Nuclear Proteins ; genetics ; metabolism ; Promoter Regions, Genetic ; physiology ; Transcription, Genetic ; physiology ; YY1 Transcription Factor ; genetics ; metabolism