Objective To study the induction of IL-6 on expression and activity of tissue factor (TF)in peripheral blood monocytes(PBMCs).Methods rhIL-6 100 ng/L and rhIL-6 100 ng/L+ rhIL-6 MoAb 10?g/L were used respectively to stimulate the PBMCs for 24 h,PBMCs without any stimulation as control group,to study the changes of antigen content,mRNA expression and the ac- tivity of the TF.Results The antigen content,mRNA expression and activity of TF were increased significantly in 100 ng/L rhIL-6 group as compared with rhIL-6 100 ng/L+rhIL-6 MoAb 10?g/L and control groups(P＜0.01).Conclusions rhIL-6 can induce the increase of antigen expression,activity and mRNA expression in PBMCs,and these effects can be suppressed successfully by rhIL-6 MoAb. This study demonstrated that there was potential relations between cytokines and thrombogenesis, which may play an important role the pathogenesis of chronic rejection.

Laboratory animals and animal experiments are foundations and important support conditions for life sciences, especially for medical research. The animal experiments have drawn extensive attention from the society because of the ethical issue. This paper takes Wenzhou Medical University as an example to give a brief introduction to the ethical review about laboratory animals in the university so as to further draw attention and concerns from the public about the ethical issue of laboratory animals. We successively introduce its scientific projects, nurturing environment and ethical review of laboratory animals.
Animal Experimentation ; ethics ; Animals ; Animals, Laboratory ; Universities

ObjectiveTo investigate the variations and distributions of the plasmid-mediated quinolone resistance genes in clinical isolates of Shigella and their resistance to antimicrobial agents. Methodsqnr, aac(6')-Ib-cr and qepA genes were identified by polymerase chain reaction (PCR) in 137 clinical isolates of Shigella.DNA sequencing of gene-positive strains were analyzed and the conjugation experiment was performed. The minimal inhibitory concentrations (MIC) of Shigella isolates, recipient strains and transconjugants were tested by agar dilution method for quinolones and other antimicrobial agents. The genotype of transconjugants were determined by PCR and sequencing. ResultsFour (2.9％) strains of the 137 Shigella isolates were qnr gene positive, including 3 qnrS2 positive and 1 qnrB4 positive (GenBank accession numbers of the complete sequence were JF261185 and HQ917003, respectively).Furthermore,five (3.6％) aac ( 6')-Ib-cr gene-positive strains (GenBank accession number JF261186 ) and one (0.7％) qepA gene-positive strain were identified in all isolates. The conjugation experiments were successfully carried out in six out of ten PCR-positive isolates. The MIC of transconjugants against quinolones and other antimicrobial agents increased differently compared to recipient strains. Conclusions The plasmid-mediated quinolone resistance genes are lowly prevalent in clinical isolates of Shigella. However, these resistance genes have the characteristic of horizontal transfer, which indicates that more attention should be paid to this phenomenon.

Objective To evaluate the clinical significance of the diameter of the esophageal hiatus on multislice spiral CT(MSCT)and to present the MSCT manifestations of esophageal hiatus hernia (EHH).Methods(1)The distance between diaphragmatic crura(DDC),which indicated the diameter of esophageal hiatus,was measured in 140 normal adult patients on their thoracic and/or abdomenal CT images.(2)The DDC of 56 patients with EHH diagnosed by barium examination was measured on MSCT, and the MSCT findings were analyzed retrospectively.Results(1)The DDC of 140 normal adult cases were(13.44?4.41)mm on average and increased with age.The mean DDCs of patients under the age of 59 year-old(80 cases)and over 60-year-old(60 cases)were 11.03?2.10 mm and 16.67?4.64 mm respective]y,there was a significant difference(t=8.762,P

Objective This study aimed at exploring the causative genes and summarizing the clinical characteristics in two Chinese families with thoracic aortic aneurysm and dissection ( TAAD ) .Methods The whole exome capture and high throughput sequencing were applied to identify the causative gene.Family members were examined for features of syndromic ge-netic diseases by clinician and geneticist.Results Four known TAAD candidate genes were identified in family TAA01:rs140598(FBN1), rs185661462(MYH11), rs77620762(MYLK3), and rs111426349(TGFBR1).The TGFBR1 mutation (c.1459C>T) had been confirmed to co-segregate with the TAAD phenotype in all affected family members.Early onset of aortic root dilatation was significant in this family , and the average age at diagnosis of aortic root dilatation or aneurysm was23. 2 years.ACTA2(c.445C>T) was proved in family TAA02, and livedo reticularis was confirmed.Conclusion The causa-tive genes were identified via whole exome capture and high throughput sequencing in two TAAD families .Early onset of aortic root aneurysm was proved in TAA01, while livedo reticularis was found in TAA02.

ObjectiveTo analyze the clinical distribution and antimicrobial resistance profile of Serratia marcescens(S. marcescens), and to provide the scientific evidence supporting clinical diagnosis and treatment.MethodsThe antimicrobial susceptibility test was performed in 104 strains of S. marcescens by agar dilution method. The results were judged according to the criteria recommended by Clinical and Laboratory Standards Institute (CLSI) 2010.The data were analyzed by chi square test. Results The majority of S. marcescens were isolated from sputum specimens,accounting for 59.6％ (62/104). The bacteria were most frequently isolated from department of respiratory (33.7％,35/104),followed by intensive care unit (23.1％,24/104),department of gerontology (16.3％, 17/104). The results of antimicrobial susceptibility test showed that the resistance rates of S.marcescens against ampicillin,gentamicin and cephazolin were high,which were 90.4％,86.5％ and 79.8％,respectively; those against the 3rd generation of cephalosporins were 24.0％-43.3％. No imipenem and meropenem resistant strains were identified. Compared with cefoxitin-resistant strains,the resistance rates of non-cefoxitin resistant strains against piperacillin (82.9％ vs 28.6％),ceftazidime (63.4％ vs 9.5％),aztreonam (68.3％ vs 9.5％),amikacin (68.3％ vs 20.6％),ciprofloxacin (48.8％ vs 19.1％) and chloramphenicol (90.3％ vs 58.7％) were all lower (all P ＜ 0.05 ). Conclusions S. marcescens is one of the most common conditional pathogenic bacteria leading to nosocomial infections,which is resistant to many kinds of antimicrobial agents.The surveillance of antimicrobial resistance in S. marcescens should be strengthened for purpose of preventing the transmission of multidrug resistant strains.

Objective Clinical significance of using ELISA to determine ?-amyloid(A?)_(1-42) antibody levels in the sera of patients with Alzheimer's disease(AD).Methods 96 wells PVC plate was coated with A?_(1-42)peptide.Serum of AD patient was competing with mouse A?_(1-42)monoclonal antibody in this assay.The second antibody was horseradish peroxidase(HRP)conjugated goat anti-mouse IgG.Serum A?_(1-42)antibody levels were determined by ELISA.Results The sensitivity of this assay was about 1 ng/ml.The recovery rate of this test was between 96.5% and 104.7%.The residual A?_(1-42)antibody levels in human serum or horse serum after A?_(1-42)antibody was removed by absorption were less than 1 ng/ml. Serum A?_(1-42)antibody levels in 37 AD patients[(5.1?1.9)ng/ml]were remarkably lower than those in normal people[(12.6?3.3)ng/ml,P

S-adenosylmethionine-dependent uroporphyrinogen III methyltransferase (SUMT) is a novel red fluorescence indicator. However, the production of SUMT in Escherichia coli is restricted by its relatively low solubility, and little is known about the red fluorescent materials that are associate with SUMT. Two individual SUMT mutations, L166A and L88R/L89G double mutant were produced by site-directed mutagenesis. Both mutants were overexpressed in E. coli and purified by Ni-NTA chromatography. The reddish mixtures isolated from the purified L88R/L89G double mutant were analyzed by UV-visible spectra scanning and mass analysis(MS). The L88R/L89G double mutant has enzymatic activity in vivo, whereas L166A mutant loses the activity. Trimethylpyrrocorphin is identified as the main constituent in the isolated pigments. The purified L88R/L89G mutant increases protein solubility, which is applied potentially as the fluorescent indicator denoting the solubility of protein fusion partner.
Amino Acid Substitution ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; Fluorescence ; Mass Spectrometry ; Methyltransferases ; chemistry ; genetics ; metabolism ; Molecular Weight ; Mutagenesis, Site-Directed ; methods ; Mutation ; Pigments, Biological ; chemistry ; metabolism ; Plant Proteins ; chemistry ; genetics ; metabolism ; Recombinant Proteins ; chemistry ; metabolism ; Solubility ; Spectrophotometry, Ultraviolet ; Zea mays ; enzymology ; genetics

OBJECTIVETo explore the application value of morphology assessment of sperm from fresh semen in routine in vitro fertilization (IVF).

METHODSWe analyzed the morphology of the sperm from fresh or optimized semen samples and, based on the sperm morphology of the raw semen, allocated 908 IVF cycles due to the pure tubal factor to different groups: morphologically normal sperm (MNS) ≤ 4%, > 4% - ≤ 15%, and > 15% in Trial 1 and MNS ≤ 1%, > 1% - ≤ 2%, > 2% - ≤ 3%, and > 3%-- ≤ 4% in Trial 2. We compared the rates of fertilization, cleavage, high-quality embryo, -blastocyst formation, and pregnancy among different groups.

RESULTSThe total fertilization rate was significantly lower in the MNS ≤ 4% than in the MNS > 4% - ≤ 15% and >15% groups (74.40% vs 78.61% and 80.03%, P < 0.01). Compared with the MNS ≤ 1%, > 1% - ≤ 2%, and > 2% - ≤ 3% groups, the MNS > 3% - ≤ 4% group showed remarkably increased rates of 2PN normal fertilization (77.23%, 78.97% and 78.99% vs 85.47%, P < 0.01), cleavage (95.71%, 96.01% and 97.27% vs 98.73%, P < 0.05), and blastocyst formation (53.85%, 49.01% and 49.55% vs 63.41%, P < 0.01). No statistically significant differences were observed in the rates of clinical pregnancy, implantation, early abortion, live birth, or malformation at birth among different groups (P > 0.05).

CONCLUSIONMNS ≤ 4% affected the total rate of fertilization while MNS ≤ 3% reduced the rate of normal fertilization in IVF. However, even MNS ≤ 1% did not result in fertilization disorder or failure. Therefore, teratozoospermia alone was not an indicator of ICSI and sperm mor- phology assessment had no obvious value for predicting the rates of embryo quality, clinical pregnancy, and live birth in IVF.

Objective To analyze the result of treatment for acute-on-chronic liver failure patients with fast high efficiency Nucleoside and to explore the relations among inhibition to virus replication , liver failure development and immune rejection .Methods Sixty-two cases of acute-on-chronic liver failure pa-tients with HBV DNA(+) were divided into study group (treated with a kind of fast and nucleoside , n =30) and control group( n =32).HBV DNA,CD4 +T,CD8 +T, C3,C4 TBIL,PTA were observed at treat-ment 0w,2w and 4w.Results All of the study and control group patients , serum HBV DNA were positive before treated.And the levels of CD4+,CD8 +,C3,C 4,TBIL,PTA of study group were not significantly compared with control group .At treatment 2w , the rate of HBV DNA diverted negative in study group 90.0%(27/30), was significantly more then control group (9.4%, 3/32)(χ2=37.14 , P <0.01).But the CD4 +,CD8 +,C3,C4,TBIL,PTA levels were not significantly however between study and control group . At treatment 4w ,the rate of HBV DNA diverted negative in study group (96.7%, 29/30), was significant-ly more then control group(12.5%,4/32) (χ2 =40.74, P <0.01).CD4 +, CD8 +,C3,C4,TBIL,PTA levels of the study group were significantly more compared with the control group .The CD4 +level of study group (495.33 ±89.91)cells/ml, was higher significantly then control group (270.34 ±97.74)cells/ml( t=9.42, P <0.01),the CD8 +level (571.03 ±120.15 ) cells/ml, was higher significantly then control group(224.88 ±79.68)cells/ml( t =13.45, P <0.01).The C3 level of the study group (0.28 ±0.11) g/L, was lower significantly then control group ( 0.68 ±0.13 ) g/L ( t =13.13 , P <0.01 ) , the C4 level (0.12 ±0.06)g/L, was lower significantly then control group (0.23 ±0.10)g/L( t =4.92, P <0.01). The TBIL level of study group ( 653.93 ±131.02 )μmol/L, was higher significantly then control group (285.63 ±154.63)μmol/L( t =10.09, P <0.01),the PTA level (17.13 ±7.07)%, was lower signifi-cantly then control group(50.94 ±13.68)%( t =12.10, P <0.01).The death rate of the study group( 57.9%) was higher significantly compared with the control group (28.1%)(χ2 =6.39, P <0.05).Con-clusion Treatment of chronic severe hepatitis with fast and high efficiency nucleoside may arise the T cell subset level and make the immune rejection strength , as a result the liver failure maybe far away from cure .