Objective To prepare recombinant human adenovirus type 3 expressing Norovirus cap-sid protein gene(Noro-orf2). Methods The cDNA for Noro-orf2 was amplifed by RT-PCR from stool of in-fantile gastroenteritis and cloned into the adenovirus shuttle vector pBSE3CMV-egfp. The vector pBSE3CMV-Nor was linearized with EeoR Ⅴ and Not Ⅰ, and transformed into E. coil BJ5183 with lined edenovirus ge-nomic DNA pLasmid pBRAdv3 by Rsr Ⅱ. The identification of recombinant adenovirus plasmid pBRAdv3E3dNor was performed by PCR, enzyme digestion and DNA sequencing. Then pBRAdv3E3dNor was digested with AsiS Ⅰ and transfeeted into Hep-2 cells with LipofectAMINETM 2000 to package recombi-nant adenovirus particles. Results Noro-orf2 was successfully inserted into the shuttle vector. The recombi-nant adenoviral plasmid pBRAdv3E3dNor was generated by homologous recombination in E. coil BJ5183 and confirmed by PCR and enzyme digestion. The recombinant adenovirus was successfully packaged and puri-fied. Norovirus eapsid protein gene expression was confirmed in Hep-2 cells by immunecytochemistry assay. Conclusion The recombinant type 3 adenovirus expressing Norovirus eapsid protein gene was successfully constructed. This study laid a foundation for developing vaccine against Norovirus.

Objective To analyze the relationship between the genetic polymorphisms of uridinediphosphate glucuronosyltransferase 1A9 (UGT1A9) and mycophenolic acid (MPA)pharmacokinetics in Chinese kidney recipients.Methods Gene mutations (C-440T/T-331C,C-2152T,T-275A,T98C) were detected in 196 recipients by PCR-LDR.On the 28th day after transplantation,the plasma samples which were obtained at the time points of predose,0.5 h and 2 h after administration were measured by an immunoassay method (Emit Mycophenolic Acid Assay,Dade Behring).MPA-AUC0-12 was calculated based on these three data.Correlation between single nucleotide polymorphisms (SNPs) and MPA pharmacokinetics was analyzed.Results C-2152T,T-275A and T98C genotypes of UGT1A9 were not found in 196 recipients.The frequency of C-440T/T-331C gene mutation was 14.29% (28/196).The mean value of MPA-AUC0-12 was 40.6±11.8 wild genotype,respectively (P＞0.05).Conclusion C-2152T,T-275A and T98C genotypes of UGT1A9 are scarce in Chinese kidney recipients.In this study,there is no distinct relationship between -440/-331 SNPs and MPA pharmacokinetics in Chinese allograft recipients.

Increased recognition of human parvovirus B19,as a significant human pathogen has resulted in intensive researches to understand the pathogenesis of B19 infection,to elucidate the nature of Th1-mediated cellular immune response,to improve diagnostic strategy that is deployed to detect B19 infection and blood-product contamination,and to lay a foundation that should contribute to the development of an effective vaccine to prevent B19 infection.In this review,the biologic characteristics and the pathogenesis of human parvovirus B19,and B19-related manifestations as well as laboratory diagnostic methods for B19 infection were comprehensively discussed.

This paper analyses the data structure of DICOM standard by the applications in the Non-DICOM format fluoroscopic images converted into the DICOM format images. It puts forward a solution to integrate the Multi-Channel Electrophysiology Recorder System with the X -ray system in the cardio-catheter room.
Computer Communication Networks ; Diagnostic Imaging ; instrumentation ; methods ; Electrophysiologic Techniques, Cardiac ; instrumentation ; Fluoroscopy ; instrumentation ; Humans ; Image Processing, Computer-Assisted ; instrumentation ; methods ; standards ; Information Storage and Retrieval ; Radiology Information Systems ; standards ; Software Design ; Video Recording ; instrumentation ; methods ; standards

OBJECTIVETo study the oncogene transcriptor c-myc, stimulatory protein 1 (SP1) expression in ameloblastoma (AB) and their relation with telomerase reverse transcripase (hTERT), and to investigate the clinical biological characteristics of AB.

METHODSThe expression was observed in AB by in situ hybridization and SP method.

RESULTSThe positive rates of c-myc mRNA, hTERT mRNA and SP1 protein were 81.5% (44/54), 94.4% (51/54) and 83.3% (45/54), respectively. Their positive rates increased as AB recurred and transformed malignantly. A strong correlation was found between hTERT and c-myc, hTERT and SP1 (rs = 0.853, P < 0.001; rs = 0.900, P < 0.001).

CONCLUSIONActivity of telomerase plays an important role in the tumorigenesis development of AB. Increasing of hTERT expression may be related to c-myc and SP1. The expression of these three parameters has a significant correlation with the clinical biological characteristics of AB.

Ameloblastoma ; metabolism ; Humans ; Proto-Oncogene Proteins c-myc ; metabolism ; Sp1 Transcription Factor ; metabolism ; Telomerase ; metabolism

OBJECTIVETo study the major metabolites of antitumor lead compound T-OA (oleanolic acyl-3, 5, 6-trimethyl pyrazine-2-methyl ester) in rat urine, in order to preliminarily infer its metabolic mode in rats.

METHODRat urines of the blank group, the raw material group (ligustrazine TMP and oleanolic acid OA Moore equivalent) and the T-OA group were collected and freeze-dried; Solids were extracted by ethyl acetate; And then the extracts were re-dissolved with acetonitrile. HPLC-HRMS coupling technique was adopted to find the potential mass spectrum peak under ESI(+) (see symbol) ESI(-) modes. Metabolite-related information was obtained by comparing the three groups of spectra.

RESULTOne metabolite of OA and two metabolites of TMP were identified in the raw material group; none metabolite of T-OA but one phase II metabolite was detected in the T-OA group.

CONCLUSIONIt is the first time to identify one phase II metabolite of T-OA and one phase II metabolite of OA were identified in rat urine. On that basis, the researchers preliminarily inferred that T-OA does not show the efficacy in the form of raw material. The HPLC-HRMS method established could be used to identify metabolites of related derivative structures. This paper could also provide certain reference for designing pro-drugs based on oleanolic acid.

Animals ; Antineoplastic Agents ; chemistry ; metabolism ; urine ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; metabolism ; Male ; Mass Spectrometry ; methods ; Molecular Structure ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the DNA methylation of human telomerase reverse transcriptase (hTERT) promoter in ameloblastoma (AB) and investigate its clinical biological significance.

METHODSDNA methylation of hTERT promoter in 12 cases of AB and 11 cases of normal oral mucosa were detected by methylation-specific PCR.

RESULTSThe positive cases of unmethylated hTERT promoter region in AB and normal oral mucosa were 4 (4/12) and 6 (6/11), respectively (P < 0.001). The positive cases of methylated hTERT promoter region in AB and normal oral mucosa were 11 (11/12) and 3 (3/11), respectively. Four cases of AB and 1 case of normal oral mucosa showed both DNA methylation and unmethylation of hTERT promoter region in the same time.

CONCLUSIONSDNA methylation of of hTERT promoter region in AB was more common than that in normal oral mucosa. The DNA methylation in hTERT promoter maybe regulate hTERT gene expression.

Adolescent ; Adult ; Aged ; Ameloblastoma ; genetics ; DNA Methylation ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Jaw Neoplasms ; genetics ; Male ; Middle Aged ; Promoter Regions, Genetic ; genetics ; Telomerase ; genetics ; Young Adult

OBJECTIVETo investigate the expression of cyclin E mRNA, p21(WAF1) mRNA and p27(KIP1) protein in human ameloblastoma (AB), and to explore the clinical and biological characteristics of AB.

METHODSThe expression of cyclin E mRNA, p21(WAF1) mRNA and p27(KIP1) protein in 54 cases of human AB were detected by in situ hybridization or immunohistochemistry (SP method).

RESULTSThe positive expression rate of cyclin E mRNA in the cytoplasm or cell nucleus of AB was 66.7% (36/54). The expression of cyclin E mRNA increased with AB recurrence and malignant transformation, and the difference of expression among primary AB, recurrent AB, and malignant AB, was statistically significant. The positive expression ratio of cyclin E mRNA in OKC was 50.0% (8/16). The p21(WAF1) mRNA expression in the cytoplasm or cell nucleus of AB decreased, and the positive ratio was 22.6% (12/54) in AB, 37.5% (6/16) in OKC, respectively. The p27(KIP1) protein expression in the cell nucleus of AB was positive in a small number of cases, and the positive rate was 16.7% (9/54) in AB, 6.3% (1/16) in OKC, respectively.

CONCLUSIONSThe genesis and invasion of AB is associated with the cell proliferation and differentiation, and regulated by the higher expression of cyclin E and the lower expression of p21(WAF1) and p27(KIP1).

Adolescent ; Adult ; Aged ; Ameloblastoma ; metabolism ; pathology ; Child ; Cyclin E ; genetics ; metabolism ; Cyclin-Dependent Kinase Inhibitor p21 ; genetics ; metabolism ; Cyclin-Dependent Kinase Inhibitor p27 ; Female ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; metabolism ; Jaw Neoplasms ; metabolism ; pathology ; Middle Aged ; Oncogene Proteins ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Young Adult

OBJECTIVETo study the expression of HOXC13 mRNA in ameloblastoma (AB), and to investigate its biological significance.

METHODSHOXC13 mRNA was examined in 47 cases of AB (primary AB 29 cases, recurrent AB 14 cases, malignant AB 4 cases). 2 cases of fibrous dysplasia of bone, 10 cases of keratocystic odontogenic tumor (KCOT) and 7 cases of normal oral mucosa were selected as control.

RESULTSThe positive rates of HOXC13 mRNA in AB, KCOT, and normal oral mucosa were 97.9% (46/47), 7/10 and 3/7, respectively. There was a significant difference among AB, OKC and normal mucosa (chi(2) = 21.665, P = 0.001). For HOXC13, the keratinizing cells and granulizing cells in AB were negative, some fibroblasts were positive, 2 cases of fibrous dysplasia of bone were positive.

CONCLUSIONSHOXC13 was highly expressed in AB. The expression of HOXC13 mRNA in AB had heterogeneity, which could improve the epithelial proliferation, and its loss may lead to the cornification and degeneration of epithelial cells.

Adolescent ; Adult ; Ameloblastoma ; genetics ; metabolism ; pathology ; Female ; Gene Expression ; Genes, Homeobox ; Homeodomain Proteins ; metabolism ; Humans ; Male ; Middle Aged ; Mouth Mucosa ; metabolism ; Odontogenic Tumors ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Young Adult

AIM To study the effects of Danggui Buxue Decoction total glycosides combined with Hirudo in the treatment of pulmonary fibrosis in rats and its mechanism.METHODS One hundred and twenty male SD rats were selected,eighteen of which were taken as normal group (normal saline),the remaining 102 rats were used to establish model for pulmonary fibrosis.Seventy-two modeled rats were randomly divided into model group (normal saline),Danggui Buxue Decoction total glycosides group (13 mg/kg),Hirudo group (Hirudo powder,4 g/kg) and combination group (Danggui Buxue Decoction total glycosides,13 mg/kg;Hirudo powder,4 g/kg),eighteen rats in each group with intragastric administration.RESULTS On the 7th,14th and 28th days after modeling,alveolar inflammatory score,pulmonary fibrosis degree score,TGF-β1 and PAI-1 expression levels in lung tissue,IL-4 and IL-17 expression levels in alveolar lavage fluid,hydroxyproline (HYP) content in lung tissue in the model group were significantly higher than those in the normal group (P ＜ 0.05).Alveolar inflammatory score,pulmonary fibrosis degree score,TGF-β1,PAI-1,IL-4,IL-17 expression levels and HYP content in the Danggui Buxue Decoction total glycosides group,Hirudo group and combination group were significantly lower than those in the model group (P ＜ 0.05).In the combination group,alveolar inflammatory score,pulmonary fibrosis degree score,TGF-β1,PAI-1,IL-4,IL-17 expression levels and HYP content were significantly lower than those in the Danggui Buxue Decoction total glycosides group and Hirudo group (P ＜ 0.05).CONCLUSION Danggui Buxue Decoction total glycosides combined with Hirudo have good therapeutic effects on pulmonary fibrosis in rats mainly by reducing TGF-β1,PAI-1 expression levels and HYP coment in lung tissue.