Breast Neoplasms ; genetics ; pathology ; Chromosome Aberrations ; Chromosome Deletion ; Chromosomes, Human, Pair 1 ; Chromosomes, Human, Pair 17 ; Chromosomes, Human, Pair 9 ; Female ; Genome, Human ; Humans ; Nucleic Acid Hybridization ; methods ; Phyllodes Tumor ; genetics ; pathology

OBJECTIVETo investigate the effect of effective parts of Zingiber officinal (EPZ) on the adhesion of ECV-304 cells with monocytes cultivated in vitro and on the expression of monocyte chemotactic protein-1 (MCP-1) and adhesion molecules.

METHODThe model of ECV-304 cell oxidative stress injury was established by hydrogen peroxide (H2O2). Then EPZ-contained blood serum was taken as experimental drug. The adherence of monocytes to endothelial cell were measured by method of rose Bengal. The total RNA of cells was extracted. The intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and MCP-1 mRNA expression in cells were detected by RT-PCR. MCP-1 protein expression were detected by ELISA.

RESULTEPZ could decrease the adhesion of monocytes with ECV-304 cells obviously. Meanwhile it could diminish the expression of ICAM-1, VCAM-1 and MCP-1 in injured ECV-304 cells.

CONCLUSIONEPZ could inhibit H2O2-induced ICAM-1, VCAM-1 and MCP-1 expression in ECV-304 and could inhibit the adherence of monocytes to endothelial cell, which may result in the protect effect in endothelial cells.

Animals ; Cell Adhesion ; drug effects ; Cell Line ; Cells, Cultured ; Chemokine CCL2 ; biosynthesis ; genetics ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Endothelial Cells ; cytology ; drug effects ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Ginger ; chemistry ; Humans ; Hydrogen Peroxide ; pharmacology ; Intercellular Adhesion Molecule-1 ; biosynthesis ; genetics ; Monocytes ; cytology ; drug effects ; metabolism ; Plants, Medicinal ; chemistry ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction ; Vascular Cell Adhesion Molecule-1 ; biosynthesis ; genetics

OBJECTIVETo investigate whether the connection of p27(Kip1) to S-phase kinase-associated protein 2 (Skp2) plays an oncogenic role in intraductal proliferative lesions of the breast.

METHODSHere we investigated the mechanism involved in association of Skp2’s degradation of p27(Kip1) with the breast carcinogenesis by immunohistochemical method through detection of Skp2 and p27(Kip1) protein levels in 120 paraffin-embedded tissues of intraductal proliferative lesions including usual ductal hyperplasia (UDH, n=30), atypical ductal hyperplasia (n=30), flat epithelial atypia (FEA, n=30), and ductal carcinoma in situ (DCIS, n=30). Moreover, the expression status of Skp2 and p27(Kip1) in 30 cases of the normal breast paraffin-embedded tissues were explored.

RESULTSThe DCIS group was with the highest Skp2 level and the lowest p27(Kip1) level, and the UDH group was with the lowest Skp2 level and the highest p27(Kip1) level.Both Skp2 and p27(Kip1) levels in the DCIS group were significantly different from those in the UDH group (all P<0.01).The levels of Skp2 and p27(Kip1) in the FEA group were significantly different from both the DCIS and UDH groups (all P<0.05).p27(Kip1) was negatively correlated with Skp2 in both the UDH group (r=-0.629, P=0.026) and DCIS group (r=-0.893, P=0.000).

CONCLUSIONOverexpression of Skp2 might be the mechanism underlying p27(Kip1) over degradation.

Adult ; Aged ; Breast ; pathology ; Breast Neoplasms ; etiology ; Carcinoma, Intraductal, Noninfiltrating ; etiology ; Cell Proliferation ; Cyclin-Dependent Kinase Inhibitor p27 ; physiology ; Female ; Humans ; Hyperplasia ; Middle Aged ; S-Phase Kinase-Associated Proteins ; physiology

OBJECTIVETo improve the mini-modified semi solid rappaport vassiliadis most probable number (mini-MSRV MPN) method for Salmonella detection.

METHODSBased on the mini-MSRV MPN method,Buffered Peptone Water (BPW) was modified as one step enrichment medium and Modified Semi Solid Rappaport Vassiliadis (MSRV) medium was ameliorated as modified MSRV for Salmonella detection under standard Salmonella addition recovery. A total of 154 raw chicken samples, 48 swabs of pheasantry and 48 poultry dung samples were collected to compare the detection results of Salmonella by using improved mini-MSRV MPN, mini-MSRV MPN and regular most probable number (MPN) method.

RESULTSSalmonella recovery was < 2.7 MPN/g when the standard Salmonella addition was at the concentration of 0.9 CFU/g when the mini-MSRV MPN method was employed. If the standard Salmonella addition were at 9.0 and 90.0 CFU/g, the recoveries of bacteria were 10.1 and 94.0 MPN/g, and the average recovery rate was 112% and 104%, respectively. Salmonella detection rate of modified mini-MSRV MPN, mini-MSRV MPN and regular MPN method was 18.4% (46/250), 5.2% (13/250) and 6.0% (15/250), respectively. The detection rate was higher for modified mini-MSRV MPN method than of the other two methods (χ(2) values were 19.68 and 17.82, respectively, all P values < 0.05). The detection quantity of Salmonella (medians were 21.0, < 2.7 and < 3.0 MPN/g, respectively). The quantity detected by modified mini-MSRV MPN method was higher than that of the other two methods (both Z values were 5.71, both P values < 0.05).

CONCLUSIONModified mini-MSRV MPN method is an accurate method for foodborne Salmonella detection.

Animals ; Chickens ; microbiology ; Culture Media ; Food Contamination ; analysis ; Food Inspection ; methods ; Salmonella ; isolation & purification

OBJECTIVETo analyze the allergenicity and immunogenicity of Psilogramma menephron allergen so as to provide the basis for preparing recombinant and standardized allergen vaccines of Psilgramma menephorn.

METHODSThe extracts of Psilgramma menephorn were analyzed by SDS-PAGE, and the allergenicity and immunogenicity of the extracts were tested with 9 sera from allergic patients by means of immunoblotting.

RESULTSMore than 20 allergen proteins were separated from the extract of Psilgramma menephorn by SDS-PAGE, with the relative molecular weight ranging from 12,000 to 128,000. The relative molecular weight of the allergenic proteins were 74,000 (88.9%), 66,000 (22.2%), 49,000 (22.2%), 36,000 (77.8%), or 25,000 (33.3%), and those of the immunogenic proteins were 79,000 (33.3%), 74,000 (66.7%), 66,000 (22.2%), 49,000 (22.2%), 36,000 (44.4%), or 25,000 (55.6%).

CONCLUSIONThe relative molecular weight of the major allergenic proteins of Psilgramma menephorn are 74,000 and 36,000, and 74,000 and 25,000 for the major immunogenic proteins. These proteins constitute the major allergenic components for diagnosis and specific treatment of Psilgramma menephorn allergy.

Adolescent ; Adult ; Allergens ; immunology ; isolation & purification ; Animals ; Asthma ; blood ; immunology ; Blotting, Western ; Female ; Humans ; Immunoglobulin G ; blood ; Lepidoptera ; immunology ; Male ; Middle Aged ; Young Adult

Vascular endothelial growth factor-D (VEGF-D) and vascular endothelial growth factor receptor-2, -3 (VEGFR-2, -3) with their corresponding signaling pathway play significant roles in the development of the embryonic vascular system and pathological lymphangiogenesis. The study was aimed to express and purify the GST-VEGF-D fusion protein, and to explore the angiogenesis effect of VEGF-D. The total RNA was extracted from human fetal lung tissue, and the mature form of VEGF-D was expanded by polymerase chain reaction (PCR), then the plasmid pGEX-5X-1/VEGF-D was reconstructed and the GST-VEGF-D fusion protein expressed in transformed E.coli BL21-DE3. The results showed that the molecular mass of this fusion protein was 38 kD and compassed more than 15% of the total bacteria proteins. The fusion protein was recognized by anti-GST and anti-VEGF-D antibodies. The soluble GST-VEGF-D fusion protein could interact with VEGFR-3/Fc and was able to stimulate the proliferation of human erythroleukemia cell line (HEL) cells. The data of chick embryo chorioallantoic membrane (CAM) experiments indicated that GST-VEGF-D could induce the CAM angiogenesis. It is concluded that the GST-VEGF-D fusion protein with biological activity was successfully expressed, and which may provide an experimental model for the investigation of the VEGF-D-induced angiogenesis and lymphangiogenesis.
Animals ; Chick Embryo ; Chorioallantoic Membrane ; blood supply ; Humans ; Neovascularization, Physiologic ; drug effects ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology ; Vascular Endothelial Growth Factor D ; biosynthesis ; genetics ; pharmacology

AIMThe process of vascular calcification involves various genetic alterations which may play a very important role in the vascular calcification. Vascular smooth muscle cells undoubtedly composed the main part of vascular cells, and are involved in vascular calcification. So bovine artery smooth muscle cell (BASMC) was used to investigate the gene changes during BASMC's calcification.

METHODSBovine artery smooth muscle cells cultured in vitro was induced calcified by beta-Glycerophosphate (beta-GP). Using DD-PCR technique to screening differentially expressed genes and those differentially expressed bands were reexamined by reverse Northern blot. All the ESTs were sequenced and BLAST with GenBank.

RESULTSTotal 65 cDNAs were isolated as differentially expressed genes and 40 of them were successfully reamplified. Using reverse-Northern blot, seven of these 40 cDNAs were reproducibly expressed differentially between the two cells. Three of them are new bands and have not been reported before.

CONCLUSIONThis is the first time using DD-PCR to screen differentially expressed genes of BASMC calcification. Seven related ESTs were identified relating to BASMC calcification.

Animals ; Arteriosclerosis ; genetics ; metabolism ; pathology ; Cattle ; Cells, Cultured ; Expressed Sequence Tags ; Genetic Variation ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; metabolism ; pathology ; Vascular Calcification ; genetics ; metabolism ; pathology

OBJECTIVETo explore F (13) A gene mutation in a pedigree with hereditary coagulation factor XIII (FXIII) deficiency.

METHODSThe FXIII deficiency was diagnosed by clot solubility test and other standard laboratory clotting tests. All exons, exon-intron boundary sequences of F(13) A gene were amplified by PCR and the products were sequenced directly. Any mutation identified by direct sequencing was confirmed by reverse sequencing. The mutation identified in the proband was screened in the family members.

RESULTSThe assays of PT, Qiulan, fibrinogen leveling, platelet counts, bleeding time were normal and the clot solubility test was positive in the proband. The homozygous deletion of 33 nucleotides (127067de133) in exon 10 of F(13) A gene which resulted in deletion of 11 amino acids in FXIIII A protein with 720aa residues was identified in the proband. Family studies showed that the mutation was inherited from the parents both of whom carried the heterozygous deletion mutation.

CONCLUSIONThe homozygous 127067de133 mutation of F(13) A gene is responsible for the disorder of the pedigree.

Adolescent ; Factor XIII ; genetics ; Factor XIII Deficiency ; genetics ; Heterozygote ; Homozygote ; Humans ; Male ; Pedigree ; Sequence Deletion

OBJECTIVETo study clinical features, treatment and curative effects in children with acute clenbuterol poisoning, in order to provide a basis for early diagnosis and treatment.

METHODSClinical data of 28 hospitalized children with acute clenbuterol poisoning in April 2011 were retrospectively studied.

RESULTSOf the 28 patients, there were 15 males and 13 females, aged 1 to 13 years (mean age 6.5±4.8 years). Vomiting, palpitations and limb shaking were found as main clinical manifestations in the patients. Main changes of blood biochemical included hypokalemia, lactic acidosis, hyperglycemia, hypsocreatinkinase. Snus tachycardia and S-T segment depression were observed on ECG. Patients' symptoms were gradually alleviated after 12-78 hours by use of beta blockers, potassium supplement, protecting the heart and other symptomatic and supportive treatment. Blood biochemical indexes were improved after 48 hours of admission. All of the patients were cured after 5 days. The symptoms of the patients do not longer occur during a follow up of half a month.

CONCLUSIONSAcute clenbuterol poisoning is characterized by vomiting, palpitations, limb shaking, hypokalemia, lactic acidosis and tachycardia in children. An early effective treatment of this disease can improve prognosis in children.

Acute Disease ; Adolescent ; Adrenergic beta-Agonists ; poisoning ; Child ; Child, Preschool ; Clenbuterol ; poisoning ; Electrocardiography ; Female ; Humans ; Infant ; Male ; Retrospective Studies

Objective To study the family rearing pattern of attention deficit hyperactivity disorder(ADHD)with or without anxiety disorder and to explore its risk factors.Methods 9495 children and their parents were sampled at random in Hunan province,using two-stage investigation.Those who were diagnosed ADHD and the normal control filled out Egna Minnen av Barndoms Uppfostran and family adaptability and cohesion scale bv themselves.Results The comparison of factors as:actual family cohesion,parents' punishments,reiection,mother's excessive protection,intervention and father's excessive protection were significantly difierent between ADHD with or without anxiety disorder and normal children(P＜0.05).The comparison of parents' punishments,reiection,excessive protection and intervention were obviously different between ADHD with anxiety disorder and simple ADHD(P＜0.05).Mother's reiection was the influencing factor of simple ADHD,with OR as 1.122.Ideal family cohesion,mother's rejection and father's punishments were the influencing factors of ADHD with anxiety disorder,with OR as 0.966.1.215 and 1.089 respectively.Conclusion There were some problems in the parental rearing pattern of ADHD with or without anxiety disorder.Mother's rejection,father's punishments and ideal family cohesion were suggested to be correlated with ADHD and anxiety disorder.