Adolescent ; Catheterization ; adverse effects ; Endocarditis, Bacterial ; etiology ; Female ; Heart Septal Defects, Ventricular ; therapy ; Humans ; Postoperative Complications

ObjectiveTo explore the relationship between the negative co-inhibitor programmed death-1 ( PD-1 ) and the pathogenesis of myasthenia gravis ( MG), by detecting the expression of PD-1 and programmed death ligand-1 ( PD-L1 ) on peripheral blood mononuclear cells (PBMCs) and soluble PD-1 (sPD-1) in plasma from myasthenia gravis patients. MethodsPeripheral blood samples were collected from 45 MG patients and 33 healthy persons without prednisone or other immunodepressant treatment during the half year ahead of withdrawal.The expression of PD-1 and PD-L1 on PBMCs were detected using immuno-fluorescence labeling and flow cytometry, and the concentrations of sPD-1 in plasma were measured using an ELISA kit. Results(1) The proportion of CD4+ PD-1 + T cells, as well as CD14+ PD-L1 +monocytes of the MG group was higher than that of the control group. There were no significant differences in the proportion of CD4+ PD-1 + T cells or CD14+ PD-L1 + monocytes in the MG sub-groups between different genders or MG types. While the proportion of CD4+ PD-1 + T cells of the late-onset MG (age ≥40) group was higher than that of the early-onset MG group (age ＜40). And it was higher in the MG patients with thymoma or thymus hyperplasia than that from the MG patients with normal thymus. The proportion of CD14+ PD-L1 +monocytes from the MG patients with thymoma or thymus hyperplasia group decreased obviously compared with that of the patients with normal thymus group; but no difference could be found between the late-onset group and early-onset group. (2)The concentration of sPD-1 in the plasma from the group of MG patients was(6. 92 ±0. 72) ng/ml,which was higher than that of the healthy control group ( (3.28 ±0. 42) ng/ml),even more, it was significantly higher in the early-onset MG group than that of the late-onset MG group,there was a negative correlation( r =-0. 526, P =0. 000) between the age of onset and the concentration of sPD-1. ConclusionsThe increased expressions of PD-1 on CD4+ T cells and PD-L1 on CD14+ monocytes in MG patients suggested the involvement of the couple of molecules in the pathogenesis of MG.Higher concentration of soluble PD-1 in the plasma of patients with MG suggested that it might disturb the ligation of PD-1 and PD-L1 on T cells and antigen presenting cells, which might result in the abnormal transportation of the negative modulating signal, and accelerate the pathological progress of MG.

Background Age-related cataract is one of the common causes of blindness.Although the pathophysiology of age-related cataract is far from clearly understood,it is well accepted that DNA damage plays an important role in the disease pathogenesis.Objective The purpose of this study was to quantitatively evaluate the DNA damage in peripheral lymphocytes of age-related cataract.Methods A cross-sectional study was carried out.This study complied Declaration of Helsinki and approved by Ethic Committee of Affiliated Hospital of Nantong University.Written informed consent was obtained from each subject.Two hundred and eleven patients with agerelated cataract and 147 normal subjects were enrolled from a “ Jiangsu Eye Study:Funing 2011 Eye Disease Epidemic Survey”.All the subjects aged from 50 through 80 years with matched age and gender between the two groups.The percentage of tail DNA and Olive tail moment (OTM) were detected by comet assay to assess the extent of DNA damage in peripheral lymphocytes.Statistical analyses were performed with SPSS 17.0 software,and the differences of the percentage of tail DNA and OTM were compared between the age-related cataract group and normal control group by independent sample t test as well as among the 50-59 years group,60-69 years group and ≥70 years group by one-way analysis of variance.Results Comet assay showed a round lymph cell with the clear border in the normal group;while in the age-related cataract group,the cell was bigger with a comet-like tail.The percentage of tail DNA and OTM in peripheral lymphocytes were (21.75 ± 3.51) ％ and 6.54 ± 1.65 in the age-related cataract group,and those in the normal control group were (9.31 ±3.60)％ and 2.18 ± 1.10,respectively,with significant differences between them (t =32.67,P =0.00 ; t =28.02,P =O.00).In the 50-59 years subgroup of the age-related cataract group,the percentage of tail DNA and OTM in peripheral lymphocytes were (20.04±2.86) ％ and 5.92± 1.14,and in the 60-69 years subgroup of the age-related cataract group,the percentage of tail DNA and OTM in peripheral lymphocytes were (20.77 ±2.93) ％ and 6.13 ± 1.14,which were significantly reduced in comparison with (22.79 ± 3.67)％ and 6.95±1.91 of the ≥70years subgroup(TailDNA％:q=2.75,P=0.00; q=2.02,P=0.00;OTM:q=1.03,P =0.02 ; q =0.82,P =0.00).Conclusions The pathogenesis and development of age-related cataract probably is associated with DNA damage.

OBJECTIVELong time exhaled oxygen will induced oxygen toxicity. Some studies had found that different pathology may exised in normobaric and hyperbaric pulmonary oxygen toxicity, and nitric oxide synthase (NOS) may play a role. In this study, we discussed the change of NOS in normobaric and hyperbaric pulmonary oxygen toxicity.

METHODSSixty male SD rats were randomly divided into 6 groups (n = 10), exposed to 1 ATA (atmosphere absolute), 1.5 ATA, 2 ATA, 2.5 ATA and 3 ATA, 100% oxygen for 56, 20, 10, 8, 6 hours respectively. Rats were exposed to air as control. After exposure, the protein in bronchoalveolar lavage fluid (BALF), the wet/dry weight of lung and the expression of eNOS, nNOS in lung were defined.

RESULTSAs compared to air group, the protein in BALF, the wet/dry of lung were significantly elevated in 1.0 ATA group, while these changes were not so obviously in the other groups, and these changes in hyperbaric oxygen group (approximately 1.0 ATA) were significantly decreased as compared with nonnrmobaric oxygen group (1.0 ATA). The expression of nNOS were not changed in normobaric and hyperbaric pulmonary oxygen toxicity, while the expression of eNOS was significantly decreased in 2 ATA group, and significantly elevated in 2.5 ATA and 3 ATA group.

CONCLUSIONThe expression of eNOS can change when exposed to different pressures of oxygen.

Animals ; Disease Models, Animal ; Lung ; metabolism ; Male ; Nitric Oxide Synthase Type I ; metabolism ; Nitric Oxide Synthase Type III ; metabolism ; Oxygen ; poisoning ; Pressure ; Rats ; Rats, Sprague-Dawley

Objective To explore the regional parameters of triple screening model of Down syndrome in the second trimester in Jinhua City.Methods A total of 20 232 second trimester pregnant women with single fetus (gestational age at 15 -20 +6 weeks)was enrolled,and their serum samples were determined by American Perkin Elmer company Auto DELFIA automatic time -resolved fluorescence immunoassay analyzer for Down syndrome screening with triple markers, namely AFP,free β-hCG and uE3 .The risks of Down syndrome were evaluated by Lifecycle 3.2 software.And the risks of Down syndrome were re -calculated by local statistical median equations.Pregnant women were suggested to receive amniotic fluid fetal karyotype analysis if the risk of Down syndrome were equal or above 1 /270.Results Local median marker levels were significantly higher than the software built -in median levels (P <0.01).Both true -positive detection rates (sensitivity)were 87.50%.The false positive rate of local median equations was 4.24%,while the built -in median equations was 4.74%.Conclusion There are significant differences on the race and region by using the LifeCycle 3.2 median equations.The local equations may lower the false positive rate.

OBJECTIVETo investigate the expression of extracellular matrix metalloproteinase induced (EMMPRIN) in the interface tissue, and explore the role of EMMPRIN in the aseptic loosening of prostheses.

METHODSImmunohistochemistry was performed to characterize the EMMPRIN-expressing cells at sites of interface tissue around aseptic loosened hip prostheses in 16 cases. Reverse transcription-polymerase chain reaction (RT-PCR) was performed to study the existence of EMMPRIN mRNA in interface tissue samples. And it was followed up by computer assisted image analysis in order to detect the A values of their expression. Synovium of hip joint of 8 femoral neck fracture were in control group.

RESULTSStrong immunostaining of EMMPRIN was found in the macrophages and fibroblasts of lining-like layers and vascular endothelium of synovial membrane-like interface tissue around loosened prostheses. Expression of EMMPRIN was significantly higher in interface tissue than the control synovium (z=-3.252, P=0.001). RT-PCR of interface tissue samples disclosed the presence of EMMPRIN mRNA of 14 cases. In interface tissue, the A value of EMMPRIN increased significantly compared to control synovium (P<0.01).

CONCLUSIONOver-expression of EMMPRIN up-regulates the production of matrix metalloproteinase (MMPs) in the interface tissue. And it can promote the bone destruction around prostheses. Thereby it may be one of methods to prevent and treat aseptic loosening of prostheses by repression the biology activity of EMMPRIN.

Adult ; Aged ; Arthroplasty, Replacement, Hip ; Basigin ; genetics ; physiology ; Female ; Hip Prosthesis ; Humans ; Immunohistochemistry ; Male ; Matrix Metalloproteinases ; metabolism ; Middle Aged ; Osteolysis ; physiopathology ; Prosthesis Failure ; RNA, Messenger ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Synovial Membrane ; metabolism

OBJECTIVESTo explore a sensitive and specific method for detection of bcl-2/IgH gene rearrangement in diffuse large B cell lymphoma (DLBCL), and verify the credibility of the established method.

METHODSbcl-2/IgH hemi-nested PCR primers were designed using the professional primer design software. Fifty-two samples of pathologically diagnosed DLBCL and 10 fresh tonsil tissues were amplified using hemi-nested touch down-PCR to detect bcl-2/IgH gene rearrangement. The PCR products were cloned and sequenced.

RESULTSbcl-2/IgH gene rearrangement was detected in 6 of 52 DLBCL samples and 2 of 10 fresh tonsil tissues using one-way method. By using the hemi-nested PCR for the second round amplification, 5 of DLBCL were positive, but all of the fresh tonsil tissues were negative. The positive PCR products were sequenced and analyzed on the Internet, 3 of 8 cases obtained by one-way method were false positive, 5 positive cases amplified using hemi-nested PCR were all bcl-2/IgH gene rearrangement. PCR products of 3 false positive cases were homologous to BAC331191 and LLNLR-245D11 in human chromosome 19 and RP11-498P10 in chromosome 1.

CONCLUSIONThere are false positive results using common primers for detecting bcl-2/IgH gene rearrangement. The mechanism may be that highly homologous sequences to human genome exist in commonly used primers. The specificity of the diagnosis could be improved by hemi-nested PCR using the combination of primers we designed and the traditional ones.

Gene Rearrangement, B-Lymphocyte, Heavy Chain ; Genes, bcl-2 ; genetics ; Humans ; Lymphoma, Large B-Cell, Diffuse ; genetics ; Polymerase Chain Reaction ; methods

X-linked sideroblastic anemia (XLSA) is caused by mutations of erythroid-specific 5-aminolevulinate synthetase (ALAS2) gene. In this study a eukaryotic expression vector of ALAS2 was constructed and transfected into eukaryotic cells to observe the expression of ALAS2 gene. The full length cDNA of ALAS2 gene was inserted into plasmid pDs-red2-N1, named pDs-red2-N1/ALAS2. Then, the vector was transfected into K562 cells via electroporation. At 48 hours after transfection, total RNA from K562 cells was extracted, expressions of ALAS2 gene and protein with red fluorescence in the K562 cells were detected by RT-PCR and flow cytometry, respectively. The vector was also transfected into COS 7 cells via liposome. Both mRNA and protein expression in COS7 cells were detected by RT-PCR and fluorescence microscopy. The result showed that after the pDs-red2-N1/ALAS2 eukaryotic expression vector was digested by KpnI and BamHI, two fragments of 4 700 bp and 1 764 bp were displayed by electrophoresis on agarose gel. Sequence method confirmed that the sequence was correct. RT-PCR amplified the total RNA extracted from the transfected K562 and COS7 cells, and could find mRNA of ALAS2 gene that can't be found in K562 and COS7 cells usually. The expressions of both fluorescein and ALAS2 were significantly increased. The percentage of positive cells reached about 19.2% and 10.7%, respectively. ALAS2 expression lasted for 10 days in COS7 cells and the peak was at the third day. It is concluded that the eukaryotic expression vector of ALAS2 gene is successfully constructed; K562 and COS7 cells transfected with the vector via electroporation and liposome can express ALAS2 protein. So, the vector has the potential in gene replacement and can be used for patients with XLSA in future.
5-Aminolevulinate Synthetase ; genetics ; Anemia, Sideroblastic ; genetics ; therapy ; Animals ; COS Cells ; Chromosomes, Human, X ; Genetic Linkage ; Genetic Therapy ; Genetic Vectors ; Humans ; K562 Cells ; Microscopy, Fluorescence ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo study the association between the polymorphism of human monoamine oxidase type A (MAO-A) gene and Parkinson's disease(PD).

METHODSFnu4HI restriction fragment length polymorphism(RFLP) and PCR-RFLP were used to detect the mutation of MAO-A gene. The frequencies of alleles and genotypes at the MAO-A Fnu4HI locus on the X chromosome in different PD group were compared with those of the control group.

RESULTSIt was found that the frequencies of G allele in the patients with PD and controls were 0.613 and 0.527 respectively, P=0.039 "the frequencies of TT genotype were 0.303 and 0.415(P=0.014), and the frequencies of GG genotype were 0.564 and 0.451 respectively(P=0.021). When the patients were divided into two groups by age-onset, significant difference in the allelic and genotypic frequencies was observed only between early-onset PD group and control group. And when the PD patients were grouped by sex, significant difference was observed only between male PD group and male control group (the frequencies of G allele being 0.669 and 0.500 respectively, P=0.005).

CONCLUSIONThis study revealed significant differences between PD group and control group in allelic and genotypic frequencies. The findings supported the hypothesis about an association between MAO-A gene and PD, suggesting that age at onset of PD and gender predisposition might be related to the putative association, and Fnu4HI SNP be a risk factor for PD.

Alleles ; Asian Continental Ancestry Group ; Deoxyribonucleases, Type II Site-Specific ; analysis ; genetics ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Male ; Monoamine Oxidase ; genetics ; Parkinson Disease ; genetics ; Polymorphism, Genetic ; genetics ; Polymorphism, Restriction Fragment Length

OBJECTIVETo investigate the DNA and chromosome damage in peripheral blood lymphocyte of workers occupationally exposed to formaldehyde (FA).

METHODSAll 151 workers occupationally exposed to FA from two plywood factories and 112 workers without occupational FA exposure working in a machine manufactory were recruited into this study. Comet assay and cytokinesis-block micronucleus technique was used to evaluate the DNA and chromosomal damage of peripheral blood lymphocyte. The air FA samples were collected with SKC 224-PCXR8 air samplers. Gas chromatography was used to analyze the FA level. Personal information including occupational history, age, sex, smoking and drinking status was collected by the questionnaire.

RESULTSThe time weighted average concentration (TWA) of FA in the working environment of FA-exposed workers (range 0.10 - 7.88 mg/m(3)) was higher than those in controls (< 0.01 mg/m(3)). The olive tail moment (Olive TM) in low FA-exposed workers [3.03 (2.49 - 3.67)] was lower than that in high FA-exposed workers [3.95 (3.53 - 4.43)], but higher than that in controls [0.93 (0.78 - 1.10)], the differences were statistical significant (P < 0.05). Comet trail length in FA-exposed workers were significantly higher than that in controls [6.78 (6.05 - 7.60)], but no significant differences ware found between the high FA-exposed workers [12.59 (11.80 - 13.43)] and the low FA-exposed workers [11.25 (10.12 - 12.50)]. The frequency of micronuclei per 100 binucleated cells in low FA-exposed workers (0.41 +/- 0.25) was lower than that in high FA-exposed workers (0.65 +/- 0.36), but higher than that in controls (0.27 +/- 0.13), the differences were statistical significant (P < 0.05). The increased tendencies with the exposure levels were found in those three indices. In stratification analysis, the same results were found.

CONCLUSIONIn the current FA exposure levels, the DNA and chromosomal damage in peripheral blood lymphocyte might be induced by FA exposure, and be increased with the levels of exposure.

Adult ; Alcohol Drinking ; Comet Assay ; DNA Damage ; Formaldehyde ; analysis ; poisoning ; Humans ; Lymphocytes ; drug effects ; metabolism ; Micronucleus Tests ; Occupational Exposure ; analysis ; Smoking ; Young Adult