Objective To construct eukaryotic expression vector of human pancreatic duodenal homeobox 1(PDX-1) gene,and to detect its expression in NIH3T3 cell lines. Methods The whole coding sequence of PDX-1 gene was amplified by polymerase chain reaction(PCR) from human pancreatic-cell tumors cDNA.The fragment was inserted into eukaryotic expression vector pcDNA3.1 plasmid.The recombinant plasmid was verified by double digestion and DNA sequencing.The expression of PDX-1 gene in NIH3T3 cells was assayed by Western blot. Results The length of specific fragment amplified by PCR was 852 bp,and the recombinant plasmid pcDNA3.1-PDX-1 showed two bands of 5.5 kb and 852 bp by digestion using respective restriction enzymes BamHⅠand EcoRⅠ.The sequence of PDX-1 gene was approved or confirmed by blasting to GenBank.It was suggested that PDX-1 gene had been cloned into pcDNA3.1 vector correctly.Western blot showed that PDX-1 gene was expressed,which was detected 24 h after pcDNA3.1-PDX-1 plasmid was transfected into NIH3T3 cells. Conclusion The recombinant eukaryotic expression vector pcDNA3.1-PDX-1 was successfully constructed and expressed in NIH3T3 cell lines.

Objective To obtain abundant human betacellulin(BTC) with biological activity. Methods The whole mature protein coding sequence of BTC gene was amplified by polymerase chain reaction(PCR) method applied to human pancreatic ?-cell tumors cDNA.The fragment was cloned into prokaryotic expression vector pET32a(+) plasmid.The recombinant plasmid was transformed into E.coli BL21 and the fusion protein was expressed under isopropyl-beta-D-thiogalactopyranoside(IPTG).The fusion protein was purified by Ni2+ affinity chromatography.SDS-PAGE and Western blot were employed to determine the expression and purification of the expected protein.BTC was added to culture NIH3T3 cells for 5 days,and cell proliferation was detected by MTT. Results Lots of fusion protein were produced,and the purified protein can stimulate the proliferation of NIH3T3 cells. Conclusion The human BTC can be successfully obtained from the pET32a(+) system with the biological activity of stimulating the proliferation of NIH3T3 cells.

No abstract available.

No abstract available.
Humans ; Irritable Bowel Syndrome*

Aneurysm, Dissecting ; surgery ; Aorta, Thoracic ; surgery ; Humans ; Male ; Middle Aged ; Stents

OBJECTIVE To investigate the distribution and resistance of fungi isolated from hemopathy patients,and to provide the laboratorial data in order to prevent and treat infection caused by fungi more effectively.METHODS Strains were isolated from the upper respiratory tract,lower respiratory tract,genitourinary tract,alimentary tract;and perianal region,perineum,and the soft tissue and skin wounds.Antimicrobial susceptibility of clinical isolates was tested by broth microdilution susceptibility test.RESULTS Candida albicans,Aspergillus,tropicalis,C.krusei,and C.glabrata were the main fungi of total 3104 detected isolates.The resistance rates of C.albicans and C.tropicalis to fluconazole,itraconazole and 5-flucytosine were no more than 1.4%.The MIC50-MIC90 range of C.tropicalis for itraconazole was narrow(0.25-0.5 mg/L),but that was wide for fluconazole(0.25-4.0 mg/L).CONCLUSIONS C.albicans,Aspergillus,C.tropicalis,C.krusei,and C.glabrata are possibly the most common pathogens causing endogenous and exogenous fungi infection.The main infection sites of Aspergillus are the upper respiratory tract and lower respiratory tract,which may be associated possibly with ward's air quality.The MIC50to MIC90 range of C.tropicalis is wider for fluconazole than for itraconazole,which is possibly correlated with overexpression of CtMDR1 in C.tropicalis caused by the widely application of fluconazole in therapy.

Objective To investigate the function of middle-methoxyl pectin-Ca2+ complex in(Graduate School,Chinese Academy of Science,Beijing 100039;1 Polymer Engineering Laboratory,Changchun Institute of Applied Chemistry,Chinese Academy of Sciences,Changchun 130022,China) lowing postprandial serum glucose of normal and impaired glucose tolerance(IGT) rats.Method The experimental IGT rats models were made by injecting STZ into abdomen.Then normal and IGT rats were given test meal and their postprandial serum glucose were measured.Results The middle-methoxyl pectin-Ca2+ complex could significantly improve the glucose tolerance and weaken the peak value of postprandial serum glucose of normal and IGT rats(P

Leiomyoma is one of the rarest solid tumor of the ovary. Approximately 50 cases have been published to date. However, most reported leiomyoma of the ovaries were small and rarely induced serious symptoms. We report a case of ovarian leiomyoma in 57-year-old woman which has been experienced in our haspital with brief review of literature.
Female ; Humans ; Leiomyoma* ; Middle Aged ; Ovary*

Objective To construct a lentiviral vector containing mir-7-3 gene and green fluorescent protein (GFP) gene,and to detect the expression of mir-7-3 gene in U251 cells.Methods The fragments containing all the mir-7-3 gene were amplified by RT-PCR and were cloned into the lentivirus vectors labeled with GFP,which was transfected together with the packaging plasmids into 293T cells by CaC12.The supernatant was collected,concentrated,identified,and was transfected to U251 cells of gliomas.Fluorescent microscopy was used to observe the fluorescence in the 293T cell,and real time RT-PCR was used to examine the relative contents of mir-7-3 in U251 cells.Results Electrophores was shown that the sequence of the RT-PCR product was consistent with the data of mir-7-3 by DNA sequence analysis,indicating that the mir-7-3 gene was successfully cloned,and strong green fluorescence was observed by fluorescent microscopy.The supernatant of lentivirus-transfected 293T cells effectively infected U251 cells and the relative content of mir-7-3 was observed in the transfected U251 cells.Conclusion It is concluded that the lentiviral vector containing mir-7-3 gene was constructed successfully,which provides a basis for further study of mir-7-3 function.

No abstract available.
Arthritis, Juvenile*