[Association between ZNF365 gene polymorphisms and bronchial asthma in children].

OBJECTIVE: To study the association of single nucleotide polymorphisms (SNP) (rs2393903 and rs10995251) in ZNF365 gene with bronchial asthma and its clinical characteristics in Han Chinese children in Hubei, China. METHODS: A total of 221 children with bronchial asthma and 243 normal children, all of whom were from Hubei, were recruited to carry out a case-control study. The genotype and allele frequencies of two SNPs in ZNF365 gene were determined using the polymerase chain reaction-restriction fragment length polymorphism technique. RESULTS: There were no significant differences in the distribution of three genotypes (GG, GA, AA) and allele frequency in SNP rs2393903 between the asthma and control groups (P>0.05). However, there were significant differences in the distribution of three genotypes (CC, CT, TT) and allele frequency in SNP rs10995251 between the asthma and control groups (P<0.05); C allele was a risk factor (OR=1.380). The asthmatic children with CC genotype of SNP rs10995251 had a significantly higher serum level of total immunoglobulin E (IgE) than those with TT genotype (P<0.05). CONCLUSIONS: The SNP rs10995251 in ZNF365 gene is associated with the susceptibility to bronchial asthma in children in Hubei, China, and the SNP may affect the level of serum IgE in these children.

[Evaluation of immune responses and related patho-inflammatory reactions of a candidate inactivated EV71 vaccine in neonatal monkeys].

OBJECTIVE: To evaluate the safety of enterovirus type 71 (EV71) inactivated vaccine (human diploid derived) for infection prevention in an animal model by investigating the immune responses and related patho-inflammatory reactions. METHODS: In the neonatal monkey model for EV71 vaccine protection, vaccinated group (n = 4) and unvaccinated group (n = 4) were attacked with live virus at the same time, the parameters of clinical observations, antibodies and inflammatory factors in peripheral blood and cerebrospinal fluid (CSF) were detected. And the pathological changes in major organs were used to determine the patho-inflammatory reactions during the immune responses elicited by vaccination. RESULTS: The neutralizing antibodies of vaccine group reach to 1:32. There was no obvious changes of inflammatory factors in peripheral blood and CSF of monkeys challenged or unchallenged by live virus. In peripheral blood of unvaccinated group, the level of basophilic granulocyte higher 4 - 5 times than normal level and the interferon-γ (IFN-γ) showed obvious increase. Live virus infected after 7 days, the interleukin-6 (IL-6) and IFN-γ in peripheral blood of unvaccinated group (18.5, 12.7 pg/ml) were higher than vaccinated group (10.2, 7.6 pg/ml). Furthermore, the IL-6 in CSF (102.0 pg/ml) had 4 - 5 times increased than vaccinated group (12.4 pg/ml) at 7 days after virus exposure. Meanwhile, the pathological analysis revealed that no obvious changes were detected in CNS and other organs of vaccinated monkeys challenged with live virus. However, the pathological damages induced by virus infection could be determined in the unvaccinated control monkeys, including neuronal damage, massive cellular infiltration associated with pulmonary edema/hemorrhage and pulmonary/bronchial damage due to an infiltration of inflammatory cells. CONCLUSION: Capable of inducing an immune response, the EV71 inactivated vaccine offers protection to neonatal rhesus monkeys against the attacks of live virus. Based on the results of no patho-inflammatory reaction and pathological damage after viral infection in vaccinated animals, the excellent safety of this vaccine may be confirmed in neonatal monkey.

Combination of phosphorylated and truncated EGFR correlates with higher tumor and nodal stage in head and neck cancer.

Epidermal growth factor receptor (EGFR) is a target in head and neck cancer. High EGFR expression and phosphorylated EGFR predicts poor survival in head and neck cancer patients, but does not correlate with advanced stage disease. The aim of this study is to determine if clinical biological correlates are more accurate when different aspects of EGFR are evaluated in combination. We analyzed the EGFR phosphorylation, expression, and mutations in 60 primary head and neck tumors. We not only found that head and neck tumors with either truncated or activated EGFR tend to have higher tumor and nodal stage but also discovered two novel EGFR truncations.

The Ski protein negatively regulates Siah2-mediated HDAC3 degradation.

Ski acts as a transcriptional co-repressor by multiple direct and indirect interactions with several distinct repression complexes. Ski represses retinoic acid (RA) signaling by interacting with, and stabilizing, key components of the co-repressor complex, namely, HDAC3. However, little is known as to how the Ski protein can stabilize HDAC3. In the present study, we identified the Siah2 protein as a potential E3 ubiquitin ligase that mediated proteasomal degradation of HDAC3. Reciprocal co-immunoprecipitation assays further revealed that Ski interacts with Siah2. Furthermore, co-expression of the Ski protein stabilized the level of Siah2 protein. Since Siah2 regulates its own level of expression by self-degradation, the stabilization of Siah2 by Ski is an indication that Ski association leads to inhibition of Siah2 E3 ubiquitin ligase activity. Only wild-type Ski and Ski truncation mutants that were in the same complex with Siah2 could stabilize HDAC3 levels. Taken together, the results suggest that association with Ski leads to inhibition of Siah2 E3 ubiquitin ligase activity and in this way, the Ski protein inhibits Siah2-mediated proteasomal degradation of HDAC3.

Synthesis, single-crystal characterization and preliminary biological evaluation of novel ferrocenyl pyrazolo[1,5-a]pyrazin-4(5H)-one derivatives.

A series of novel ferrocenyl pyrazolo[1,5-a]pyrazin-4(5H)-one derivatives was synthesized and characterized by 1H NMR, 13C NMR, IR, HRMS and X-ray diffraction analysis. Preliminary evaluation of biological applications showed that the compounds 6c and 6f inhibit the growth of A549 cells in dosage-dependent manners through cell cycle arrest.

[Dynamic analysis of ecological footprint of Dongying City based on a modified model].

Taking the spatiotemporal heterogeneity of water-heat condition into consideration, the traditional ecological footprint (EF) model was modified with net primary productivity (NPP). In the meanwhile, water resource EF was calculated to complement the deficiency of water EF account which only included water's fishing function. The EF dynamics of Dongying City from 1996 to 2003 was analyzed by using the modified model. Based on traditional model, the EF of Dongying City in 1996-2003 increased from 1.766 hm2 to 2.644 hm2, and the ecological capacity (EC) decreased from 0.889 hm2 to 0.813 hm2; while based on the modified model, the EF increased from 2.819 hm2 to 3.776 hm2, and the EC decreased from 1.935 hm2 to 1.865 hm2. Comparing with that from traditional model, the ecological pressure calculated by the modified model was lesser, which suggested that to increase the utilization of water resource would alleviate the ecological pressure on the region. The modified EF model was more precise to reflect the natural resource utilization of Dongying City.

The Ski protein can inhibit ligand induced RARalpha and HDAC3 degradation in the retinoic acid signaling pathway.

Recent data has implicated the Ski protein as being a physiologically relevant negative regulator of signaling by retinoic acid (RA). The mechanism by which Ski represses RA signaling is unknown. Co-immunoprecipitation and immunofluorescence assay showed that Ski and RARalpha are in the same complex in both the absence and presence of RA, which makes Ski different from other corepressors. We determined that Ski can stabilize RARalpha and HDAC3. These results suggest that Ski represses RA signaling by stabilizing corepressor complex.

The repairing effect of a recombinant human connective-tissue growth factor in a burn-wounded rhesus-monkey (Macaca mulatta) model.

CTGF (connective-tissue growth factor) has been characterized as an extracellular-matrix-associated protein that modulates basic-fibroblast-growth-factor signalling and angiogenesis. In the present paper, the cloning of the ctgf gene from human umbilical-vein endothelial cells and expression of the protein in Escherichia coli as an N-terminal hexahistidine fusion protein is described. Recombinant human CTGF (rhCTGF) was expressed and purified so that we could investigate its effect on the proliferation of human embryo fibroblast KMB-17 and NIH3T3 cells. The results indicated not only that the protein was properly folded, but also that it had the same specific activity and stability as the native protein. Furthermore, we administered this recombinant protein in a non-human primate [rhesus monkey (Macaca mulatta)] burn-wound model and report the clinical findings and structural effects. Epitheliotrophic effects were conspicuous in wounded tissues at 10-100 ng of CTGF/cm(2), suggesting that administered rhCTGF can play a normal physiological role in wound repairing in a non-human primate model.

HTRP--an immediate-early gene product induced by HSV1 infection in human embryo fibroblasts, is involved in cellular co-repressors.

The interaction between virus and receptor is a process that mimics physiological ligand binding receptors and induces signal transduction. In the investigation of the interaction between HSV1 (Herpes Simplex virus 1) and human fibroblasts via virus binding to its receptor complex on cellular membranes, the HTRP (human transcription regulator protein), a protein encoded by an immediate-early gene of cellular response against the specific stimulation of HSV1 binding, was cloned from a cDNA library established from early gene response mRNA. The localization of HTRP expressed as a fusion polypeptide with a fluorescent protein in HeLa cells was confirmed to be the nucleus. The results of a yeast two-hybrid experiment indicated that HTRP is indeed involved in the interaction with the SAP (mSin3-associate polypeptide) complex via SAP30. A pull-down test and Western blotting in vitro, and immunoprecipitation in vivo also provided evidence in support of this result. The interaction of HTRP with SAP30 in its conserved domain implies that this protein family, as the products of immediate-early genes, comprise functional molecules involved in the transcriptional regulation of cells, which might be related to the inhibition of some cell survival genes.