Objective To construct and evaluate the real-time fluorescence quantitative polymerase chain reaction for detecting IL-2R? mRNA in ankylosing spondylitis(AS) based on TaqMan technique.Methods A pair of primers and a TaqMan probe were designed by sequence in GenBank.Total RNA isolated from the fresh peripheral blood monocytes (PBMC) of homo sapiens was amplified by the real-time FQ-RT-PCR.The product was collected by agarose gel electrophoresis and sequencing analysis then ligated with pUCm-T.The recombined plasmid was transcribed to cRNA by T7 RNA polymerase in order to prepare serial standard materials.A new method was created to quantify IL-2R? mRNA in ideal condition.Sensitivity, reproducibility and efficiency were evaluated and used, combined with sIL-2R,for clinical application of AS.Results The linear range was (7~107) cells/ml.The intra-and-inter-assay coefficient variation was 8.4% and 9.6% respectively. Recombined plasmid contained the target fragment was specific and accurate by BLAST.Standard reference was constructed successfully.The RT-PCR product in AS with HLA-B27 positive groups was higher than that with HLA-B27 negative groups and health controls (P0.05).The sensitivity of IL-2R? mRNA was 96.7%.Conclusions Real Time FQ-RT-PCR of IL-2R? mRNA is constructed successfully.This is an easy,rapid,sensitive, accurate and reliable method for quantifing IL-2R? mRNA. There is highly statistical significance, compared with sIL-2R, on the expression of IL-2R? mRNA and inflammatory states between AS and control group and effective information for administration of patients.

Objective To develop fluorescence quantitative reverse transcription polymerase chain reaction (FQ-RT-PCR) for detection and quantitation of transforming growth factor ?1 (TGF-?1) mRNA in peripheral blood mononuclear cell (PBMC) of patients with chronic hepatic disease.Methods The TGF-?1 recombined plasmid was constructed by conventional molecular biological techniques,and strand-specific cRNA standard was synthesized by T7 RNA polymerase in vitro transcription.The cRNA standard and a TaqMan-MGB probe were used for quantitation of the TGF-?1 mRNA,and the assay was evaluated.Moreover,the plasma TGF-?1 was detected by ELISA.Results Sequence analysis indicated that the recombined plasmid contains the specific 102bp fragment of TGF-?1. The FQ-RT-PCR could detect as low as 6.81 copies of standard cRNA,the linear range was from 6.81 to 6.81?10~9 copies,and the coefficient variation was 1.28%-2.27% and 2.56%-2.61% respectively in intra and inter-assay.The levels of TGF-?1 mRNA in PBMC and plasma TGF-?1 of patients with chronic hepatic disease were significantly higher than that of healthy controls(P

Objective:To develop and apply a method for real-time quantifying IL-6 mRNA.Methods:By Ficoll-Hypaque density gradient centrifugation, human peripheral blood mononuclear cells(PBMC) were isolated from whole blood treated with ethylenediaminetetraacetic acid(EDTA). Total RNA extracted from the PBMC in trizol solution was reverse transcribed to cDNA, which used as templates for fluorecent real-time quantitative PCR amplification of IL-6. PCR system consists of IL-6 primer, SBY Green Ⅰ and so on.Results:The method is wide linear range, sensitive, specific, reproducible and simple.Conclusion:The method can accurately quantify IL-6 mRNA.

Objective To study the differential proteome of kidney between lupus nephritis mouse and normal mouse.Methods The proteins of kidney were separated by two-dimensional gel electrophoresis(2-DE).The gels stained by silver were scanned by ImageScanner and analyzed by PDQuest software.Results About 573?52 and 658?43 protein spots were found in the three maps of control group and LN group respectively;the match ratio was 83% and 87% respectively.One hundred and fourteen spots were found increased that showed a two fold increase as comparing to control group.Conclusion A significant difference in protein expression of LN mouse kidney was found and may be related to the pathogenesis of LN.

Objective To evaluate the cost-effectiveness of early enteral nutrition (EEN) in elder patients undergoing gastrectomy for gastric carcinoma. Methods An outcome-based retrospective review of 52 patients who had undergone gastrectomy for gastric carcinoma between July 1999 and June 2002 was performed .There were 27 patients in the EEN group, and 25 in the TPN group. Results The mean postoperative hospital days of the EEN group was significantly less than that of the TPN group (16.3 d vs. 21.3 d, t =4.6814, P

Objective To analyze the CT findings of endobronchial spread in lung adenocarcinoma. Methods The CT findings of 15 lung adenocarcinomas or bronchioloalveolar carcinomas with endobronchial spread were reviewed,the distribution and the progression of the spread were evaluated.Results All of the primary tumors were consolidation form.The spread lesions distributed in one side of the lung or both sides along the bronchus.The pleural surface was spared.The CT findings of the spread included centrilobular nodules(n=5),tree-in-bud(n=7),acinar nodules(n=2),ground-glass opacities(n=10)and air- space consolidations(n=13)in the first CT examination.5 cases of the spread lesions only presented centrilobular nodules(single form)and 10 cases presented several appearances(complex form).All of the cases were diagnosed as tuberculosis or pneumonia,and antituberculotic or antibiotic therapy was taken with no effect.The follow-up CT scans showed progression in all cases,and the spread lesions with single form became multiple consolidations.The spread lesions with complex form deteriorated faster than the single one. Conclusion Although the CT findings of the endobronchial spread of the lung adenocarcinoma is specific, the clinical history and laboratory examination also are important for the differential diagnosis with tuberculosis and other infectious diseases.

Objective Establishing an employee satisfaction indicator system for tertiary hospitals in Beijing.Methods Evaluation indicators are built on theory research,and questionnaires are designed for survey.Such methods as principal component factor analysis,and verification factor analysis are called into play to test the construct validity and establish in the end the employee satisfaction indicators system.Results Such indicators comprise five grade-1 dimension and 34 grade-Ⅱ dimension,such as hospital management satisfaction indicator.Conclusion This self-designed scale ideally mirrors the employee satisfaction of tertiary hospitals in Beijing,proving the indicators to be objective and reasonable.

This study aims to investigate the virological impact of the stalk region and cysteine (C) in neuraminidase (NA) of influenza A/Anhui/1/05 (H5N1) and A/Ohio/07/2009 (H1N1) viruses. The NA of A/ Anhui/1/05 (H5N1), defined as AH N1, lacked 20 amino acids (including C, defined as s20) as compared with NA of A/Ohio/07/2009 (H1N1) (defined as 09N1). We deleted s20 of 09N1 to construct 09N1-s20, and inserted s20 into AH N1 to construct AH N1+s20. To investigate the impact of C on the biological function of NA, we deleted C in 09N1 to construct 09N1-C and inserted C into AH N1 to construct AH N1-C. The pseudo-type viral particle (pp) system was used to evaluate the impact of these mutants on virology. The combination of 09N1-C and 09H1 (defined as 09H1::09N1-C) showed an infectivity 8 times that of the wild type 09H1::09N1, while the infectivity of the combination of AH N1+C and AH H5 (defined as AH H5::AH N1+C) was much lower than that of the wild type AH H5::AH N1. The infectivity of the combination of 09N1-s20 and 09H1 (defined as 09H1::09N1-s20) was 4 times that of the wild type 09H1::09N1; the infectivity of the combination of AH N1+s20 and AH H5 (defined as AH H5:: AH N1+s20) was 1/7 that of the wild type AH H5::AH N1. The co-existence of 09N1-C and AH H5 displayed 6 times the infectivity of AH H5::09N1, while the infectivity of 09H1::AH N1+C was very low. Multimer analysis showed that in the wild type 09N1, the forms of NA were dimer > tetramer > monomer; the major component of NA in 09N1-C was monomer; in 09N1-s20, the forms of NA were monomer > dimer. AH N1 was mainly composed of monomer; in AH N1+s20, the forms of NA were dimer > monomer > tetramer; in AH N1+C, the forms of NA were dimer > tetramer. Deletion of C or s20 from 09N1 did not change the expression of NA. The study suggested that deletion of C from the stalk region of NA in A/Ohio/07/2009 (H1N1) increases infectivity. Insertion of C into NA's stalk region of A/ Anhui/1/05 (H5N1) significantly decreases infectivity. Cysteine deletion in the stalk region is important for the infectivity of A/Anhui/1/05 (H5N1) and A/Ohio/07/2009 (H1N1). It may interfere with the infectivity via changes in NA polymerization.
Amino Acid Motifs ; Humans ; Influenza A Virus, H1N1 Subtype ; chemistry ; enzymology ; genetics ; pathogenicity ; Influenza A Virus, H5N1 Subtype ; chemistry ; enzymology ; genetics ; pathogenicity ; Influenza, Human ; virology ; Neuraminidase ; chemistry ; genetics ; metabolism ; Viral Proteins ; chemistry ; genetics ; metabolism ; Virulence

AIM:To investigate the differentiation-inducing effects of baicalin on HL-60 leukemia cells.METHODS:The effect of baicalin on differential induction in AML cell line HL-60 was evaluated by cellular morphology,clone formation assay,CD11b and CD33 expression and NBT assay.RESULTS:Baicalin inhibited the proliferation of HL-60 cells.It enhanced the expression of CD11b on HL-60 cells,also increased the expression of CD33.HL-60 cells showed differentiation morphology after the drug treatment examined by Wright-Gimesa staining and NBT assay.CONCLUSION:Baicalin possessed differentiation-inducing effects on HL-60 cells.

To describe the unique miRNA profiles for HIV seropositive individuals and identify significantly differently expressed miRNAs, we determined the expression level of 754 miRNAs of 10 HIV seropositive individuals and 10 HIV seronegative individuals by using the Taqman low density microRNA array. BRB-Array Tool was used to conduct the significance analysis, and the DIANA online tool was used to perform the miRNA target prediction and pathway analysis. A total of 56 significantly differentially expressed miRNAs were identified by microarray between HIV seropositive and seronegative individuals. Among them, 49 miRNAs were down-regulated and 7 were up-regulated, partially overlapped with reported data. Predicted target genes were mainly involved in MAPK, TGF-beta and Wnt pathways. The results shows that miRNA profile changes in HIV-1 seropositive individuals, and the 56 differentially expressed miRNAs may play important role during HIV infection. Further studies on these miRNAs may be helpful for identify key molecules involved in HIV infection and potential diagnostic markers.
Adult ; Female ; Gene Expression Profiling ; HIV Infections ; blood ; genetics ; virology ; HIV-1 ; genetics ; physiology ; Humans ; Male ; MicroRNAs ; blood ; genetics ; Young Adult