Coronary Restenosis ; pathology ; Fibroblasts ; metabolism ; pathology ; Fibrosis

Objective To compare the humoral immunity features of acute glomerulonephritis and nephritic syndrome(NS) in children for earlier differential diagnosis and rational administration.Methods Nephelometry was used to determinate the serum level of immunoglobulin(Ig)G,IgA,IgM,C3,C4 in children by England MININEPH machine and correlated reagent. The serum level of IgE was determined by IgE enzyme linked immunosorbent assay(ELISA),assay ELISA kit of America E&ELABSINC company. Immunoturbidimetry and westergren method were used to detect anti-streptolysin O(ASO) and erythrocyte sedimentation rate(ESR),respectively.Results There were significant diffe-rence features of humoral immunity between acute glomerulonephritis and NS in children.IgG,IgA,IgE,C3,C4 had significant difference between acute glomerulonephritis children and healthy children(Pa

Objective To investigate the expression of T cell immunoreceptor with Ig and immunoreceptor tyrosine-based inhibitory domains (TIGIT) on peripheral CD4+ T and CD8+ T cells from patients with rheumatoid arthritis (RA) and its significance in order to clarify its role in the development of RA.Methods Peripheral blood samples were collected from 81 patients with RA and 33 healthy controls (HC).Expression of TIGIT on the surface of peripheral blood leukocytes was detected by flow cytometry.Differences in TIGIT expression between RA and HC groups were comparatively analyzed.Correlations of TIGIT expression on CD4+ T and CD8+ T cells with several laboratory indexes were analyzed.All data were statistically analyzed.Results (1) The percentage of TIGIT-expressing CD3+ T cells in patients with RA was significantly higher than that in HC (P<0.01).Moreover, the median fluorescence intensity (MFI) of TIGIT on CD3+ T cells was significantly elevated in patients with RA as compared with that in HC (P<0.01).No significant difference in the expression of TIGIT on B cells, monocytes or neutrophils was observed between RA and HC groups.(2) The percentages of TIGIT-expressing CD4+ T and CD8+ T cells were significantly elevated in patients with RA as compared with those in HC (P<0.01).Moreover, the MFI of TIGIT on CD4+ T and CD8+ T cells were significantly elevated in patients with RA as compared with those in HC (P<0.01).(3) The percentages of both TIGIT-expressing CD4+ T cells and TIGIT-expressing CD8+ T cells in patients with RA were positively correlated with erythrocyte sedimentation rate (ESR) (rs=0.355, P<0.01;rs=0.277, P=0.013).(4) The percentages of both TIGIT-expressing CD4+ T cells and TIGIT-expressing CD8+ T cells in patients with RA were positively correlated with rheumatoid factor (RF) (rs=0.265, P=0.017;rs=0.366, P<0.01).The MFI of TIGIT on CD4+ T cells was positively correlated with RF in RA group (rs=0.226, P=0.043).The percentage of TIGIT-expressing CD4+ T cells was positively correlated with anti-cyclic citrullinated peptide (anti-CCP) antibody in patients with RA who were positive for anti-CCP antibody (rs=0.324, P=0.012).(5) The percentage of TIGIT-expressing CD4+ T cells as well as the MFI of TIGIT on CD4+ T cells in patients with RA was positively correlated with Disease Activity Score-28 (DAS28) (r=0.232, P=0.038;r=0.343, P<0.010).Conclusion The expression of TIGIT on T cells is elevated in patients with RA and correlated with inflammatory markers, antibody production and disease activity.

OBJECTIVETo study the effect of substrate stiffness on proliferation, migration of fibroblast and integrin β(1) expression in fibroblast.

METHODSFibroblasts were inoculated on silicon substrate with stiffness of (16.2 ± 0.5), (19.8 ± 1.1), and (200.1 ± 2.6) kPa. After being cultured for 5 days or 6 days, cells were counted and cell proliferative activities (recorded as absorbance value) were assessed with methyl thiazolyl blue (MTT). After being cultured for 3 days, cell cycle was detected and proliferation index (PI) was calculated. The cell scratch test was used for determination of cell migration rate on post scratch day (PSD) 0 (the day of scratch), 1, 2, and 3. After being cultured for 2 days, the expression of integrin β(1) was determined by flow cytometry with fluorescence. Data were processed with one-way analysis of variance.

RESULTS(1) The proliferative speed and proliferative activity of fibroblasts were all increased along with the increase in substrate stiffness. PI of fibroblasts inoculated on silicon substrate with stiffness of (16.2 ± 0.5), (19.8 ± 1.1), and (200.1 ± 2.6) kPa was respectively 24.8%, 27.4%, 32.4%. On PSD 2, migration rate of fibroblasts inoculated on silicon substrate with stiffness of (19.8 ± 1.1) and (200.1 ± 2.6) kPa was respectively (91.4 ± 5.1)%, (100.0 ± 1.3)%, which were higher than that of fibroblasts inoculated on silicon substrate with stiffness of (16.2 ± 0.5) kPa [(55.8 ± 6.8)%, with F value respectively 3.5, 4.0, P values all below 0.01]. (3) The expression rate of integrin β(1) in fibroblasts inoculated on silicon substrate with stiffness of (16.2 ± 0.5) kPa was the lowest (43.22%), and that in fibroblast inoculated on silicon substrate with stiffness of (200.1 ± 2.6) kPa was the highest (81.26%).

CONCLUSIONSSubstrate stiffness may have a great effect on proliferation and migration of fibroblast during the process of wound healing and scar formation, which can be related to regulation of integrin β(1) expression.

Cell Movement ; Cell Proliferation ; Cells, Cultured ; Fibroblasts ; cytology ; metabolism ; pathology ; Humans ; Integrin beta1 ; metabolism ; Mechanical Phenomena ; Silicon

To identify whether a highly repeated GT sequence from DCA1 promoter from Dunaliella salina,which have been proved to be a salt-inducible promoter in our previous study,would be a salt-inducible regulation element,different primers were designed to amplify 6 different-length fragments of DCA1 promoter from D.salina by PCR.After these fragments were respectively inserted into the HindⅢ-BamH I sites of the vector pU?GUS,serial expression vectors containing the gus gene were generated.D.salina cells transformed with these recombinant plasmids by electroporation were grown in liquid media containing different concentrations of sodium chloride respectively.GUS enzyme activity was measured histochemically and fluorometrically.The results revealed that 3 fragments containing GT repeated sequence drove the external gus gene expression and the expression pattern of the gus gene was regulated by the concentrations of sodium chloride.Additionally,the 2 fragments without tandem GT sequence drove the gus gene expression,but the expression pattern of the gus gene wasn't regulated by the concentration of sodium chloride;Also,the upstream fragment of the tandem GT sequence wasn't able to drive the gus gene expression.In conclusion,the highly repeated GT sequence from the DCA1 promoter plays an important role in the salt-inducible regulation of DCA1 promoter from D.salina and might be a novel salt-inducible element.

Adult ; Aged ; Blood Pressure ; physiology ; Earthquakes ; Female ; Heart Rate ; physiology ; Humans ; Male ; Middle Aged

OBJECTIVETo investigate the changes of gene expression file in transitional cell carcinoma of bladder after hepatocyte cell adhesion molecule(hepaCAM) overexpression.

METHODSAffymetrix Human Genome U133 Plus 2.0 Array was used to investigate the changes of gene expression profile between adenovirus-green fluorescent protein(GFP) -hepaCAM group and GFP group in transitional cell carcinoma of bladder EJ cells.Significant Analysis of Microarray(SAM) was used to screen the differentially expressed genes, DAVID software was used to conduct gene ontology analysis and wikiPathway analysis based on the differentially expressed genes. Reverse transcription-polymerase chain reaction and Western blot were applied to verify microarray data.

RESULTSCompared with the GFP group, a total of 2469 genes were up-regulated or down-regulated by more than 2 times in the GFP-hepaCAM group. Among these genes, 1602 genes were up-regulated and 867 were down-regulated.Most of the differentially expressed genes were involved in the function of cell proliferation and cell cycle regulation. The mRNA expressions of nibrin, liver kinase B1, and cyclin D1 detected by reverse transcription-polymerase chain reaction in three different bladder cancer cell lines were consistent with the microarray data.The protein expressions of nibrin and liver kinase B1 in these three cell lines measured by Western blot were consistent with the mRNA expression.

CONCLUSIONSHepaCAM can alter the gene expression profile of bladder cancer EJ cells. The well-known anti-tumor effect of hepaCAM may be mediated by regulating the gene expression via multiple pathways.

Carcinoma, Transitional Cell ; genetics ; pathology ; Cell Cycle Proteins ; metabolism ; Cell Line, Tumor ; Cyclin D1 ; metabolism ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Genes, Tumor Suppressor ; physiology ; Humans ; Nuclear Proteins ; metabolism ; Protein-Serine-Threonine Kinases ; metabolism ; Proteins ; genetics ; physiology ; Urinary Bladder Neoplasms ; genetics ; pathology

Objective To evaluate the effect of ventilation with different positive end-expiratory pressures (PEEPs) on intracranial pressure in the patients undergoing gynecological laparoscopic surgery.Methods Sixty American Society of Anesthesiologists physical status Ⅰ or Ⅱ patients,aged 25-55 yr,with body mass index of 18-27 kg/m2,scheduled for elective gynecological endoscopic surgery,were divided into 3 groups (n=20 each) using a random number table method:routine ventilation group (group A),4 cmH2O PEEP group (group B) and 8 cmH2O PEEP group (group C).The patients were mechanically ventilated with PEEP 4 or 8 cmH2O at 5 min of head-down tilt after start of pneumoperitoneum in group B and group C.The patients were mechanically ventilated in volume-controlled mode,with tidal volume 7 ml/kg,inspired oxygen concentration 50％ and inspiratory/expiratory ratio 1 ∶ 2.Blood samples were collected from the radial artery for measurement of PaCO2 and PaO2 at 5 min after tracheal intubation (T0),5,15 and 30 min of head-down tilt (T1-3),and 5 min of the supine position after the end of pneumoperitoneum (T4).The peak airway pressure (Ppeak) was recorded,dynamic pulmonary compliance (Cdyn)was calculated,and optic nerve sheath diameter was measured using an ultrasonic apparatus at T0-4.Cognitive function was assessed at 1 day before surgery and 7 days after surgery using Mini-Mental State Examination.Results Compared with group A,Ppeak was significantly increased at T1-4,and PaO2 was increased at T2 in group B,and Ppeak and PaO2 were increased at T1-4,and Cdyn was increased at T1,2 in group C (P＜0.05).Compared with group B,Ppeak was significantly increased at T4,and Cdyn was increased at T2 in group C (P＜0.05).There was no significant difference in optic nerve sheath diameter or Mini-Mental State Examination score at each time point among three groups (P＞0.05).No patients developed cognitive dysfunction at 7 days after surgery in three groups.Conclusion Ventilation with different PEEPs causes no increase in intracranial pressure of the patients undergoing gynecological laparoscopic surgery.

OBJECTIVETo evaluate the immunostimulatory capacity of human peripheral blood dendritic cells (DCs) with Her2/neu gene transfection mediated by adeno-associated virus.

METHODSThe HLA genotypes of the breast cancer cells SK-BR-3 and MCF7 were determined, and the mononuclear cells from healthy donors with matching HLA genotype were isolated by Ficoll-Hypaque density gradient separation. The isolated cells were divided into two groups with or without transfection with the recombinant virus harboring Her2/neu gene. The cells were cultured for 7 days in RPMI 1640 medium supplemented with 10% AB human serum, GM-CSF, interleukin-4 (IL-4) and tumor necrosis factor-alpha (TNF-alpha). The mature DCs were then harvested from the cell culture and their phenotypes were identified using flow cytometry. MTT assay was employed to examine the specific killing activity of the T cells induced by the DCs.

RESULTSThe DCs transfected with the recombinant adeno-associated virus expressed CD1a, CD86 and CD83 at the rate of 98.10%, 99.42%, and 84.59%, and those without the viral transfection expressed the markers at the rate 92.69%, 98.07%, and 82.72%, respectively, showing no obvious differences in the phenotypes of the two DCs. The transfected DCs, however, showed markedly higher expression rates of CD40 and CD80 than the non-transfected DCs (61.02% vs 36.19%, and 97.61% vs 55.5%, respectively). The DCs, irrespective of the transfection, showed comparable capacities in stimulating T cell proliferation. The transfected DCs exhibited the capacity of inducing the T cells to specifically kill the target tumor cells, with the highest killing rate of (39.7-/+7.2)%.

CONCLUSIONThe immunostimulatory capacity of human peripheral blood DCs are enhanced by Her2/neu gene transfection mediated by adeno-associated virus.

Breast Neoplasms ; pathology ; Cell Line, Tumor ; Cells, Cultured ; Dendritic Cells ; cytology ; immunology ; metabolism ; Dependovirus ; genetics ; metabolism ; Genes, erbB-2 ; genetics ; Genetic Vectors ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; Humans ; Leukocytes, Mononuclear ; cytology ; Receptor, ErbB-2 ; biosynthesis ; genetics ; Recombinant Proteins ; genetics ; immunology ; metabolism ; Transfection

OBJECTIVE: To explore the effect of precursor of brain-derived neurotrophic factor (proBDNF) on cultured hippocampal neuron and its intracellular mechanism. METHODS: The hippocampal neurons were dissociated from E18 rats and cultured in neurobasal medium, and then the cells were treated with proBDNF, preBDNF(propeptide of proBDNF) and proBDNF antiserum,respectively. Thirty minutes, 1 hour or 48 hours later, the cells were stained with Nissl solution, and the immunocytochemistry methods of ELK-p (Ets domain protein), ErK2(extracellular signal regulated kinase) and c-fos were performed. RESULTS: The expressions of ELK-p, ErK and c-fos were significantly upregulated in the cultured hippocampal neurons after they were treated with proBDNF protein,and the immuno-positive staining was also obvious in some nuclei. While the endogenous proBDNF was neutralized by proBDNF antiserum treatment, the expressions of ELK-p, ErK and c-fos were downregulated and many cells showed swelling and vasoculation. The immunoreactivity in preBDNF treated cells was similar to that in normal cultured cells. CONCLUSION proBDNF plays an important role in sustaining the hippocampal neuron survival through upregulating the ELK and ErK pathways.
Animals ; Brain-Derived Neurotrophic Factor ; pharmacology ; Cell Survival ; drug effects ; Cells, Cultured ; Hippocampus ; cytology ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Neurons ; cytology ; metabolism ; Protein Precursors ; pharmacology ; Proto-Oncogene Proteins c-ets ; metabolism ; Proto-Oncogene Proteins c-fos ; metabolism ; Rats ; Signal Transduction