To detect the effect of PDZ1, domain of postsynaptic density 95 (PSD-95), on apoptosis of hippocampal neurons induced by oxygen-glucose deprivation (OGD), Sprague-Dawley rat hippocampal neurons were infected with PDZ1-viruses after 21 days of plating. Twenty-four hours after infection, cells were treated with OGD for 1.5 h, then were incubated with DAPI and apoptosis-like cells were characterized, or were collected for co-immunoprecipitation and Western blot analyses. The results showed that: (1) PDZ1 overexpression was observed in hippocampal neurons; (2) Apoptosis induced by OGD was obviously decreased in neurons overexpressing PDZ1 (P<0.05); (3) Overexpression of PDZ1 prevented the binding of GluR6 to PSD-95; (4) Overexpression of PDZ1 inhibited MLK3 and JNK1/2 activation induced by OGD. These results indicate that overexpression of PDZ1 may prevent hippocampal neurons from apoptosis induced by OGD.
Animals ; Apoptosis ; Cells, Cultured ; Culture Media ; chemistry ; Disks Large Homolog 4 Protein ; Glucose ; chemistry ; Hippocampus ; cytology ; Intracellular Signaling Peptides and Proteins ; metabolism ; Membrane Proteins ; metabolism ; Neurons ; cytology ; pathology ; Oxygen ; chemistry ; PDZ Domains ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore the effect of the drug-resistant attenuated Bacillus proteus on the protein expression of CD80 and CD86 in peripheral blood dendritic cells (DCs) of hepatitis B patients.

METHODPeripheral blood monocytes were isolated from HBV-infected patients and the DCs were separated and induced to differentiate in vitro. The expression of CD80 and CD86 proteins on cultured DCs were examined using flow cytometry.

RESULTSThe expression rate of CD80 and CD86 of chronic hepatitis B patients increased significantly (P=0.000), while the positive expression rate of CD80 and CD86 showed no obvious variation in healthy individuals (P=0.185 and P=0.118, respectively).

CONCLUSIONSBacillus proteus can significantly increase CD80- and CD86-positive cell percentage in peripheral blood DCs of HBV-infected patients, but has no such an effect in healthy individuals.

Adolescent ; Adult ; B7-1 Antigen ; metabolism ; B7-2 Antigen ; metabolism ; Bacillus ; drug effects ; immunology ; Case-Control Studies ; Child ; Dendritic Cells ; immunology ; metabolism ; Drug Resistance, Bacterial ; Female ; Gene Expression Regulation ; immunology ; Hepatitis B ; immunology ; metabolism ; Humans ; Male ; Middle Aged ; Vaccines, Attenuated ; immunology ; Young Adult

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Carcinoma, Renal Cell ; genetics ; metabolism ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Kidney Neoplasms ; genetics ; metabolism ; Male ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Middle Aged ; Transforming Growth Factor beta1 ; genetics ; metabolism ; Tumor Cells, Cultured ; Young Adult

OBJECTIVEAsthma is considered as a typical psychosomatic disease. This study aimed to investigate the temperament of asthmatic children and risk factors for asthma.

METHODSTemperamental type and dimensionality were investigated by Carry Temperament Scale in 106 children with asthma. Logistic regression analysis was used to study the risk factors for the development of asthma. One hundred and six age and sex-matched normal children served as controls.

RESULTSThere were significant differences in the adaptability, mood value and attention persistence of temperament between asthmatic patients and normal controls. Higher proportion of inter-high difficult temperamental type (17.0% vs 5.7%) and lower proportion of easy temperamental type (16.0% vs 29.2%) were found in children with asthma when compared with controls (P < 0.01). Logistic regression analysis showed that the frequency of cold between 3 and 7 years old, allergic history, idiosyncratic physique, parental history of asthma, house decoration and mood value and attention persistence of temperament were risk factors for the development of asthma.

CONCLUSIONSThere were differences in the temperamental type and dimensionality between asthmatic children and normal controls. Children with inter-high difficult temperament and suffered from the above risk factors showed a higher risk for developing asthma.

Adaptation, Psychological ; Affect ; Asthma ; etiology ; psychology ; Attention ; Child ; Child, Preschool ; Humans ; Logistic Models ; Risk Factors ; Temperament

Objective To explore the role and mechanism of forkhead box proteinO3a activates Fas ligand on renal tubular epithelial cell ( RTC ) apoptosis induced by renal ischemia /reperfusion ( I/R ) . Methods The model by clamping renal pedicles for 45 minutes follow-ing reperfusion was established.The protein expression of forkhead box proteinO3a and Fas ligand were examined by western blotting.Apoptosis of RTC was assessed by TdT -mediated dUTP nick -end Labeling (TUNEL) method and transmission electron microscopic (TEM).Renal function was assessed by biochemical automatic analyzer .Results The protein expression level of forkhead box proteinO 3a and Fas ligand was increased significantly following renal I /R at 1 h.RTC nucleus was shrinking, crushing followed renal I /R, and a significant increase in the number of TUNEL -positive cells following renal I /R was displayed com-pared with the sham group.The level of blood urea nitrogen(BUN) and serum creatinine ( Scr) was increased significantly compared with the sham group (P <0.05,P <0.01).Conclusion FoxO3a could be acti-vated during renal I /R, and then up -regulated FasL protein expression , facilitated renal tubular epithelial cell apoptosis , in turn, aggravated renal I /R injury in rats.

OBJECTIVETo investigate the expression of CD19 on childhood acute leukemia (AL) and its significance, and to provide evidence for the diagnosis and differential diagnosis as well as monoclonal antibody-targeting treatment of leukemia.

METHODSThere were 210 cases of childhood AL, of which 130 cases were male and 80 were female with a mean age of 9 years old. Using a panel of 27 fluorochrome directly labeled monoclonal antibodies, 210 samples from the patients were analyzed with CD45/SSC double parameters and multi-color flow cytometry to determine the expression of CD19.

RESULTSIn the 93 cases of B lineage acute lymphoblastic leukemia (ALL), the positive rate (98.9%, 92/93) of CD19 was significantly higher than that of the other B cell related antigens, such as CD10 (88.2%, 82/93, P = 0.003), CD20 (24.7%, 23/93, P = 0.001) and CD22 (60.2%, 56/93, P = 0.001). CD19 was expressed on all 8 cases of B/myeloid (My) hybrid acute leukemia (HAL) and 1 case of B/T HAL, but was not expressed on all 24 cases of T lineage leukemia and 5 cases of T/My HAL. In the 79 cases of acute myeloid leukemia (AML), only 5 (6.3%) cases expressed CD19. The positive rate (6.3%) of CD19 on AML was significantly lower than that on B lineage ALL (98.9%, P = 0.001). The percentage of CD19 positive cells in B/My HAL (41.6% - 88.7% with a mean of 73.8%) was significant higher than that in CD19(+)-AML (21.4% - 50.4% with a mean of 24.2%; Run Sum test, P = 0.0084). Of the 210 cases, 102 were B lineage related AL including B lineage ALL, B/My HAL and B/T HAL. In B lineage related AL, the sensitivity and the specificity of CD19 was 99.0% (101/102) and 95.4% (13/108) while the positive predictive and the negative predictive values to B lineage were 95.3% (101/106) and 99.0% (103/104), respectively. Using CD19(+) as a single reagent to diagnose B lineage, the false positive rate was 4.6% (5/108) and the false negative rate was 1.0% (1/102) with a general diagnosis index (GDI) of 94.4% [GDI = 1-(false positive rate + false negative rate)].

CONCLUSIONCD19 is continuously and stably expressed on all stages of B lineage differentiation. It is a reliable cell membrane marker for diagnosing B lineage ALL and an ideal target for antibody-targeting treatment of leukemia as well; the expression degree of CD19 can be used to distinguish B/My HAL from CD19(+)-AML; CD19 didn't express on normal myeloid cells but did on some AML cells. Therefore it can be used to detect the minimal residual disease.

Adolescent ; Antigens, CD19 ; analysis ; Child ; Child, Preschool ; Female ; Flow Cytometry ; Humans ; Infant ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; classification ; immunology

OBJECTIVETo describe the prevalence of metabolic syndrome (MS) in Jiangsu province, using IDF 2005 and ATP III 2005 definition.

METHODSMultistage cluster sampling method was used and the participants were interviewed by trained health workers under a structured questionnaire. The number of research subjects was 5888. Fasting plasma glucose (FPG), blood lipids, body mass index, blood pressure (BP) of all sample population (5888 subjects) were measured.

RESULTSThe prevalence of MS was 17.48% by IDF 2005 definition and 21.95% by ATP lII 2005 definition. Women had higher prevalence than men (P < 0.01). The prevalence of MS increased with age. Abrupt increase of the prevalence started at age of 50 in women. The prevalence varied by education, marital status, occupation and income. People from the urban and the south had a higher prevalence than from the rural and from the northern part of the country. The most common combination of MS individual components were 1) central obesity, dislipidemia and high BP; 2) central obesity, dislipidemia, high BP and high FPG.

CONCLUSIONThe prevalence of MS was high in Jiangsu province. Clustering of MS components was common. It is necessary to discuss the cut-off points of central obesity for the waist circumference diagnostic criteria of MS in Chinese population. The importance of prevention of MS should be strengthened both by health professionals and the government.

China ; Female ; Humans ; Male ; Metabolic Syndrome ; epidemiology ; Middle Aged ; Obesity, Abdominal ; epidemiology ; Prevalence ; Waist Circumference

Adolescent ; Coronary Angiography ; Coronary Vessel Anomalies ; pathology ; surgery ; Female ; Fistula ; congenital ; pathology ; surgery ; Humans ; Male ; Middle Aged

The metalloproteinases/disintegrins in the snake venom act as platelet aggregation inhibitor by an antagonism against integrin on platelet through its RGD sequence and may play other important role in cell-cell fusion, cell matrix interaction and other cellular function. Ussurin is a new metalloproteinase/disintegrin that was cloned from Gloydius ussuriensis. Poly (A+) RNA was purified from the total RNA preparation from venom gland of a single G. ussuriensis using the poly (A+) tract-mRNA isolation system. A cDNA library was constructed with the SMART PCR cDNA library construction kit. The cDNA library was screened and the positive clones were selected. The full-length cDNA of Ussurin was obtained. The cDNA encoding the Ussurin precursor has a 51bp 5'-UTR, the open reading frame of Ussurin and a 490 bp 3'-UTR, the open reading frame of Ussurin cDNA nucleotide sequence is 1434 bp and codes for 478 amino acids with a predicted molecular mass of 53.2 kD and an isoelectric point of 5.37. There is no potential N-glycosylation site in the deduced sequence region. Its deduced amino acid sequence consists of four region, a signal sequence of 18 amino acid residues, a zymogen pro-peptide of 171 amino acid residues with a cysteine switch motif (PK-MCGVT) in it, a central metalloproteinase domain of 201 amino acid residues containing a conserved zinc-chelating sequence (HEXXHXXGXXH) and a methionine-turn CIM involving zinc banding also, a space sequence between metalloproteinase domain and disintegrin domain of 15 amino acid residues with a conserved T392, T397, S400, which is specific residues of the P-II snake venom metalloproteinases, a disintegrin domain of 73 amino acid residues with a characteristic RGD region and six-disulfide bonds. Ussurin belongs to P-II class. The cDNA sequence and deduced amino acid sequence of Ussurin precursor were compared with homologous sequence in the GenBank database, the result reveals high degree of homology in sequence and organization pattern of domain with metalloproteinase/disintegrin gene family of other snake species. Compared with the alignment of amino acid sequence of metalloproteinase/disintegrin member, hypervariable regions of this member were revealed, besides they share higher homologous in the zymogen domain. It suggests that the hypervariable regions are the counterparts directly suitable for interacting with different domain of receptors, different receptors or substrates.
Amino Acid Sequence ; Animals ; Base Sequence ; Cloning, Molecular ; Disintegrins ; chemistry ; genetics ; metabolism ; Metalloproteases ; chemistry ; genetics ; metabolism ; Molecular Sequence Data ; Sequence Alignment ; Viper Venoms ; enzymology ; genetics ; Viperidae ; genetics ; metabolism

OBJECTIVELeukemia is the most common hematopoietic malignancies in children. Chemotherapy is currently the primary modality of treatment for this fatal disease. Although chemotherapy is very effective in terms of cell killing, severe side effects such as severe infections, intracranial hemorrhage etc. are frequently encountered due to its poor selective damage between normal and malignant cells or tissues. Thus, a new therapy with highly selective killing of malignant cells which leaves the normal cells unaffected is desperately desired. The aim of this study was to investigate the targeting efficacy in vitro with a new clone of anti-human CD19 antibody immunotoxin 2E8-Genistein on B lineage leukemia cell line Nalm-6 cells and its mechanisms in order to provide the evidence of target therapy on B lineage leukemia and lymphoma.

METHODS2E8-Genistein immunotoxin was generated by conjugating Mab 2E8 with a tyrosine kinase inhibitor, Genistein (Gen) via the Sulfo-SANPAH, an ultra-violet sensitive reagent. Nalm-6, a CD19+ B cell leukemia cell line, was used as target cells, while Molt-3, a CD19-T cell leukemia cell line, was taken as the negative control. The morphology of the cells was observed under the reverted reversed light microscope and the viability was checked with either trypan blue exclusion or MTT methods. Two-color flow cytometry was applied to study the mechanism of cell killing.

RESULTSAfter 24 hours of culture, 2E8-Genistein showed marked target killing on Nalm-6 cells at nine different concentrations from 20 nmol/L through 100 nmol/L with cell survival rates from (71.8 +/- 7.9)% down to (16.6 +/- 12.9)%, respectively (n = 3), which were all significantly lower than that of control group (100 +/- 13.9)% (P < 0.05). The killing effect was even more significant when the concentration was over 80 nmol/L. The growth inhibition rates of this immunotoxin on Nalm-6 cells were 82%, 84% and 94%, respectively at 24, 48 and 72 hours of culture in a time dependent manner. Significant difference was observed between the cell growth curve of Nalm-6 cultured with 100 nmol/L of 2E8-Gen and those of Nalm-6 cultured with medium (blank), PBS (negative control) or the same concentration of pure 2E8 antibody (negative control) groups (F = 152.15, P = 2.15 x 10(-7)), but there was no significant difference among the three control groups (F = 1.51, P = 0.29). When Molt-3 cells were used as target cells, the cell growth curves of Molt-3 cultured with 2E8-Gen (100 nmol/L) and with negative control of blank did not show any significant difference (F = 0.34, P = 0.59). PI/FITC Annexin V double staining analysis with flow cytometry showed that the positive rate (33.45 +/- 8.77)% of early apoptosis on Nalm-6 cells induced by 100 nmol/L of 2E8-Genistein was significantly higher than that of negative control of blank (10.44% +/- 1.28%, t = -4.39, P = 0.001) at 24 hours of culture.

CONCLUSION2E8-Genistein immunotoxin can significantly target the Nalm-6 cells in vitro in a time response manner and the apoptosis induction is involved in the course of this killing effect.

Antibodies, Monoclonal ; immunology ; pharmacology ; Antigens, CD19 ; Apoptosis ; drug effects ; Cell Line, Tumor ; Flow Cytometry ; Genistein ; immunology ; pharmacology ; Humans ; Immunotoxins ; immunology ; pharmacology ; Leukemia, B-Cell ; immunology