Human metapneumovirus (hMPV) is associated with acute respiratory tract infections (ARTI) in all age groups. However, there is limited information of genetic analysis of hMPV circulating in Beijing. To learn the characteristics of structural protein genes of human metapneumovirus circulating in children in Beijing, sequence analysis of matrix (M), small hydrophobic (SH) and attachment (G) proteins of hMPV from 2006 to 2010 was performed. Phylogenetic analysis of nucleotide sequences of 42 full length M genes, 49 SH gene and 55 G gene revealed that the hMPVs from pediatric patients were divided into sub-genotypes A2, B1 and B. There were highly conserved identities among M gene, with 7 conserved mutations of amino acids between A and B genotypes which were fairly conserved in the same genotype A or B. The amino acid identities of SH were 60.7% to 64.4% between different genotypes, 93.3% - 100% among same sub-genotype and 84.7% - 88.7% between different sub-genotypes. Use of alternative transcription-termination codon, nucleotide deletion and insertion resulted in variable length of nucleotide and deduced amino acid of G protein. Amino acid identities within same genotype ranged from 81.5% - 100%, whereas sequence identities between two genotypes ranged from 34.0% - 38.6% at the amino acid level. A new cluster of G genes in sub-genotype B2 appeared due to the same mutations and insertion of two amino acids in G protein encoding genes amplified from specimens collected from 2008 to 2010. Prediction of antigen sites of SH and G protein indicated that the variation of antigen sites between different sub-genotypes existed.
Child ; China ; epidemiology ; Genetic Variation ; Genotype ; Humans ; Metapneumovirus ; genetics ; Paramyxoviridae Infections ; blood ; epidemiology ; virology ; Phylogeny ; Retroviridae Proteins, Oncogenic ; blood ; genetics ; Viral Envelope Proteins ; blood ; genetics ; Viral Matrix Proteins ; blood ; genetics

Humans ; Lymphoma, B-Cell ; genetics ; pathology ; Oncogene Protein p55(v-myc) ; United States

OBJECTIVETo investigate the feasibility of transferring p21 gene into lung tissue with recombinant adenovirus with full-length cDNA of p21 inserted in the Wistar rat model of pulmonary hypertension (PAH) induced by left-to-right shunt, study the expression of the desired gene in vivo, find if overexpression of desired gene can inhibit pulmonary hypertension.

METHODSWith full-length cDNA of p21 transfected HEK293 cells with clonfectin, and was packed, amplified in order to obtain the high-titer recombinant adenovirus (AdCMV-p21). The infection titer was determined by TCID50 method and was diluted into 1.67 x 10(8) pfu/L. Wistar rats were randomly allocated to control group (n = 10), model group (n = 15), test group (n = 10) and test control group (n = 10). In model group and test group left-to-right shunt pulmonary hypertension was developed by using cuff technique. AdCMV-p21 was transfected into test group and test control group using tracheal inhalation. The mPAP, mRVP and RVHI were measured and compared between every two groups. The left lung was immunohistochemically stained to observe the result of transfection. The right lung was HE stained to observe morphological changes in arteria pulmonalis and calculate WT%.

RESULTSThe mRVP, mPAP and WT% in model group and test group were significantly higher than those in control (P < 0.05), which suggested that the rat model of PAH was established successfully. Brown spots in the nucleus of VSMCs of pulmonary artery were seen in test group and test control group, which indicated that AdCMV-p21 was transfected successfully. The rate of transfected cells in test group was (42.8 +/- 11.6)%, which was equal to that of test control group (P > 0.05). In test group, the mPAP was (20.06 +/- 3.40) mm Hg (1 mm Hg = 0.133 kPa), mRVP was (22.53 +/- 2.53) mm Hg, WT% was (30.8 +/- 3.5)%, which were significantly lower than those in model group (P < 0.05), but higher than those in control group and test control group (P < 0.05).

CONCLUSIONThe recombinant adenovirus could successfully carry p21 and transfect the lung tissue of PAH rat model, and full expression of p21. p21 gene could inhibit the development of PAH.

Adenoviridae ; genetics ; Animals ; Hypertension, Pulmonary ; genetics ; therapy ; Male ; Muscle, Smooth, Vascular ; Oncogene Protein p21(ras) ; genetics ; Rats ; Rats, Wistar ; Transfection

Most cases of prostate cancer become hormone refractory after 12 to 18 months of androgen deprivation therapy. The etiology of the disease is thought to be multifactorial, associated with genetic, dietary, and environmental factors. The article reviews the current situation of researches at home and abroad on the molecule mechanism of hormone refractory. It expounds the influence of the androgen receptor and its genetic mutation, apoptosis and the gene changes of p53, p21, EphB2 on prostate cancer. It is hoped to be of some directive value for the studies of prostate cancer.
Androgens ; pharmacology ; Animals ; Apoptosis ; Genes, p53 ; genetics ; Humans ; Male ; Mutation ; Oncogene Protein p21(ras) ; genetics ; Prostatic Neoplasms ; drug therapy ; genetics ; pathology ; Rats ; Receptor, EphB2 ; genetics ; Receptors, Androgen ; genetics

Proteolysis targeting chimeria (PROTAC) degrades target proteins by utilizing the ubiquitin-proteasome pathway, subverting the concept of traditional small molecule inhibitors. Among the common mutation targets of non-small cell lung cancer (NSCLC), PROTAC technology has successfully achieved the effective degradation of kirsten rat sarcoma viral oncogene homolog (KRAS), epidermal growth factor receptor (EGFR), anaplastic lymphoma kinase (ALK ) and other proteins in preclinical studies. PROTAC drugs with their unique event-driven advantages, are expected to overcome acquired drug resistance caused by small molecule inhibitors and show good therapeutic potential for undruggable targets, thereby providing a new strategy for the treatment of NSCLC.
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Carcinoma, Non-Small-Cell Lung/pathology* ; Humans ; Lung Neoplasms/pathology* ; Mutation ; Protein Kinase Inhibitors/therapeutic use* ; Proteolysis ; Proto-Oncogene Proteins p21(ras)/genetics*

CPTP1 is a nontransmembrane chicken protein tyrosine phosphatase having 92% sequence homology to the corresponding 321 amino acids of human protein tyrosine phosphatase 1B (HPTP1B). Using anti-CPTP1 antibody, we identified CPTP1-like rat PTP1 of 51 kappa Da in Rat-1 and v-src-transformed Rat-1 fibroblasts. Here we show that CPTP1-like rat PTP1 binds to p60v-src in vivo and CPTP1 also can associate with p60v-src in cell lysate of v-src- transformed Rat-1 fibroblasts. Interaction between HPTP1B-type PTPs, CPTP1-like rat PTP1 and CPTP1, and p60v-src was reduced by vanadate treatment for 13 h due to down regulation of the protein level of p60v-src in vivo. Interestingly, CPTP1-like rat PTP1 was coimmunoprecipitated with a 70-kappa Da protein which has a possibility to be tyrosine- phosphorylated by p60v-src in v-src-transformed Rat- 1 fibroblasts. These results suggest that HPTP1B- type PTPs may play an important role in p60src dependent signal pathway in eucaryotic cells.
Animals ; Blotting, Western ; Cell Line, Transformed ; Chickens ; Female ; Fibroblasts ; Oncogene Protein pp60(v-src)/*metabolism ; Phosphoprotein Phosphatase/genetics/*metabolism ; Precipitin Tests ; Protein Binding ; Protein-Tyrosine-Phosphatase/genetics/*metabolism ; Rabbits ; Rats ; Recombinant Fusion Proteins/genetics/metabolism

v-Src is a non-receptor protein tyrosine kinase involved in many signal transduction pathways and closely related to the activation and development of cancers. We present here the expression, purification, and bioactivity of a GST (glutathione S-transferase)-fused v-Src from a bacterial expression system. Different culture conditions were examined in an isopropyl beta-D-thiogalactopyranoside (IPTG)-regulated expression, and the fused protein was purified using GSH (glutathione) affinity chromatography. ELISA (enzyme-linked immunosorbent assay) was employed to determine the phosphorylation kinase activity of the GST-fused v-Src. This strategy seems to be more promising than the insect cell system or other eukaryotic systems employed in earlier Src expression.
Bacterial Proteins ; biosynthesis ; chemistry ; genetics ; isolation & purification ; Glutathione Transferase ; biosynthesis ; genetics ; isolation & purification ; Oncogene Protein pp60(v-src) ; biosynthesis ; chemistry ; genetics ; isolation & purification ; Protein Engineering ; methods ; Recombinant Fusion Proteins ; biosynthesis ; chemistry ; isolation & purification ; Saccharomyces cerevisiae ; genetics ; metabolism

OBJECTIVETo explore the effect of Buyang Huanwu decoction (BHD) drug serum on rat's in vitro cultured cerebral cortical neuron apoptosis induced by hypoxia, and on the expression of p53 and p21 genes in hypoxia process.

METHODSThe model of hypoxia neuron apoptosis was established adopting Daniel method and treated with BHD drug serum. The neuron apoptosis rate was determined by flow cytometry with propidium iodide staining, the p53 and p21 gene expression was tested by immunohistochemical method with flow cytometry.

RESULTSBHD could significantly inhibit the neuron apoptosis induced by hypoxia and down-regulate the expressions of p53 and p21 genes.

CONCLUSIONBHD shows inhibition on neuron hypoxia apoptosis and down-regulating of the p53 and p21 gene expression is one of its mechanisms.

Animals ; Apoptosis ; drug effects ; Cell Hypoxia ; Cells, Cultured ; Cerebral Cortex ; pathology ; Drugs, Chinese Herbal ; pharmacology ; Female ; Male ; Neurons ; pathology ; Oncogene Protein p21(ras) ; biosynthesis ; genetics ; Rats ; Rats, Wistar ; Serum ; Tumor Suppressor Protein p53 ; biosynthesis ; genetics

The p21 overexpression is thought to be a consequence of the p53 induced activation of the p21 gene. The immunohistochemical evaluation of p53 and p21 can be a valuable means of assessing the functional status of the p53 gene product. We examined the overexpression of p21 and p53 proteins in primary gastric lymphomas and the correlation with prognosis. A total of 32 cases of gastric lymphomas was classified into low-grade lymphomas of mucosa-associated lymphoid tissue type (n=16) and high-grade B-cell lymphomas (n=16). In low-grade lymphomas, only one case showed p53 positivity and all cases were p21-negative. In high-grade lymphomas, seven cases were p53+/p21- (44%), one case was p53+/p21+ (6%), and eight cases were p53-/p21- (50%). The p53+/p21- cases had a much lower percentage of patients sustaining a continuous complete remission state (3/7, 43%) compared with other cases (6/7, 86%). From these results, we concluded that p21 expression is rare in primary gastric lymphomas. Therefore, p53-positive lymphomas can be assumed as having p53 mutation. And combined studies of p53 and p21 may be used as a prognostic indicator in primary gastric high-grade lymphomas.
Adult ; Aged ; Antibodies, Monoclonal ; Female ; Human ; Immunohistochemistry ; Lymphoma, B-Cell/*chemistry/pathology ; Male ; Middle Age ; Peyer's Patches/chemistry/pathology ; Prognosis ; Protein p53/*analysis/immunology ; Proto-Oncogene Protein p21(ras)/*analysis/immunology ; Stomach Neoplasms/*chemistry/pathology ; Support, Non-U.S. Gov't

BACKGROUND: Cyclin-dependent kinase inhibitors (CDKI), including p21, p27 and p57 of the KIP family, are negative regulators of cell cycle progression and potentially act as tumor suppressors. The expression of p21 is induced by tumor suppressor gene p53. Mutations of p53 are common and found in various human cancers. Thus, the function of p21 as a tumor suppressor may be not retained after p53 mutation in human cancers. The aim of our study was to evaluate the tumor suppressive activity of p21 and p53 in human gastric cancer. METHODS: One hundred and two patients who underwent surgery for gastric cancer at Chonnam National University Hospital were selected retrospectively for this study. The primary selection criteria were the availability of formalin-fixed and paraffin-embedded blocks and sufficient clinical follow-up for tumor-specific survival analysis. In this study, we examined the expression of p21 and p53 in human gastric cancer tissue by immunohisto-chemistry and the correlation between their expression and clinicopathological variables. RESULTS: p21 and p53 immunoreactivities were localized in the nuclei of carcinoma cells. Positive nuclear expression of p21 and p53 was demonstrated in 63.7 and 33.3% of cancer tissues, respectively. No apparent correlation was noted between p21 and p53 expression. Negative expression of p21 correlated with advanced stage and lymph node metastasis (p=0.028 and 0.017, respectively). Moreover, negative expression of p21 correlated with poor survival (p=0.037). Positive expression of p53 correlated with depth of tumor invasion (p=0.029). However, no significant correlation could be observed between the status of p53 expression and survival. Combined analysis of p21 and p53 status showed that p21 negative and p53 positive tumors had a poorer survival than other group tumors (p=0.026). CONCLUSION: These results suggest that the status of p21 and p53 expression may help in predicting the aggressive behavior of gastric cancer. However, further studies are warranted to clarify the impact of p53 on the function of p21 as a tumor suppressor.
Adult ; Aged ; Carcinoma/genetics/*metabolism ; Female ; Gene Expression ; Genes, p53 ; Human ; Male ; Middle Aged ; Oncogene Protein p21 (ras) /genetics/*metabolism ; Prognosis ; Protein p53/genetics/*metabolism ; Retrospective Studies ; Stomach Neoplasms/genetics/*metabolism ; Survival Analysis ; Tumor Markers, Biological/genetics/*metabolism