Part of the patients with transient ischemic attack (TIA) may develop ischemic stroke.Some of them may die because of complicating cardiovascular disease.Studies in recent years have shown that the ABCD2 score has an important value in the evaluation of the prognosis of TIA.This article summarizes the source and application of the ABCD2 score,and focuses on the roles of the score in the evaluation of the prognosis of TIA.

The aim of this study is to screen out the monoclonal antibody reactive to a kind of endothelial cell line (ECV304) from SZ-1, -2, -21, -22, -51, -65, -262 and explore its function of anti-angiogenesis. The inhibitory effects of monoclonal antibody reactive to ECV304 and human lung carcinom cells (A549) adhesion and migration to extracellular matrix proteins (i.e, fibronectin and collagen IV) were studied by ELISA, the inhibition of angiogenesis in vivo was analyzed by chick chorioallantoic membrane (CAM) assays, the percentage of apoptotic cells in A549 cells was assayed by flow cytometric Annexin-V-FITC/PI dual labeling technique. The results showed that SZ-21 exhibited inhibitory effects on human umbilical vein endothelial cell line (ECV304) and pulmonary cancer cell line (A549) adhesion and migration to extracellular matrix proteins (i.e, fibronectin and collagen IV). In addition, it disrupted ongoing angiogenesis on the chick chorioallantoic membrane (CAM) model. SZ-21 also induced apoptosis of the A549 cells. In conclusion, SZ-21 inhibits angiogenesis in vivo and in vitro by blocking integrin beta(3) and inducing apoptosis. SZ-21 recognized the sequence beta(3) 28 - 35 which is far away from the RGD ligation site on GPIIIa. Integrin may interact the extracellular matrix via recognition sites other than RGD sequence.
Allantois ; blood supply ; Angiogenesis Inhibitors ; pharmacology ; Animals ; Antibodies, Monoclonal ; pharmacology ; Apoptosis ; drug effects ; Cell Adhesion ; drug effects ; Cell Line ; Cell Movement ; drug effects ; Chick Embryo ; Chorion ; blood supply ; Extracellular Matrix Proteins ; pharmacology ; Flow Cytometry ; Humans ; Integrin beta3 ; immunology ; Neovascularization, Physiologic ; drug effects ; Tumor Cells, Cultured

BACKGROUNDRNA interference (RNAi) is a potential cure for amyotrophic lateral sclerosis (ALS) caused by dominant, gain-of-function superoxide dismutase 1 (SOD1) mutations. The success of such therapy relies on the functional small interfering RNAs (siRNAs) that can effectively deliver RNAi. This study aimed to design the functional siRNAs targeting ALS-associated mutant alleles.

METHODSA modified dual luciferase system containing human SOD1 mRNA target was established to quantify siRNA efficacy. Coupled with validated siRNAs identified in the literature, we analyzed the rationale of siRNA design and subsequently developed an asymmetry rule-based strategy for designing siRNA. We then further tested the effectiveness of this design strategy in converting a naturally symmetric siRNA into functional siRNAs with favorable asymmetry for gene silencing of SOD1 alleles.

RESULTSThe efficacies of siRNAs could vary tremendously by one base-pair position change. Functional siRNAs could target the whole span of SOD1 mRNA coding sequence as well as non-coding region. While there is no distinguishable pattern of the distribution of nucleobases in these validated siRNAs, the high percent of GC count at the last two positions of siRNAs (P18 and P19) indicated a strong effect of asymmetry rule. Introducing a mismatch at position 1 of the 5' of antisense strand of siRNA successfully converted the inactive siRNA into functional siRNAs that silence SOD1 with desired efficacy.

CONCLUSIONSAsymmetry rule-based strategy that incorporates a mismatch into siRNA most consistently enhances RNAi efficacy and guarantees producing functional siRNAs that successfully silence ALS-associated SOD1 mutant alleles regardless target positions. This strategy could also be useful to design siRNAs for silencing other disease-associated dominant, gain-of-function mutant genes.

Amyotrophic Lateral Sclerosis ; genetics ; Cell Line ; Humans ; RNA Interference ; physiology ; RNA, Small Interfering ; genetics ; physiology ; Superoxide Dismutase ; genetics ; Superoxide Dismutase-1

Migraine is a complex and heterogeneous disorder. Although several genetic models has been proposed including autosomal-dominant/autosomal recessive, sex-linked, sex-limited, mitochondrial, and multi-gene, none of these models can well-explain the transmission of the disease. We hypothesied that migraine is a sex-conditioned inherited disorder (autosomal dominant in females and autosomal recessive in males). This hypothesis is supported by the evidence such as the locations of genes associated with familial hemiplegic migraine, possibly "typical" migraine as well (dominantly on chromosome 19p, 1q, and 2q), male:female ratio of prevalence (1:2-1:4), the mostly youth-onset, the provocation by the contraceptives, the induction by menstruation, and the self-limitation after menopause. Female sex-hormones appear to be the key contributor to a higher prevalence of migraine in female. Socio-environmental factors may also play an important role.
Age of Onset ; China ; epidemiology ; Female ; Genes, Dominant ; genetics ; Genetic Predisposition to Disease ; epidemiology ; genetics ; Gonadal Steroid Hormones ; metabolism ; Humans ; Inheritance Patterns ; genetics ; Male ; Menstrual Cycle ; genetics ; Migraine Disorders ; epidemiology ; genetics ; Sex Factors

AIMTo study the effects of ferulic acid (FA) on E-selectin expression in human umbilical vein endothelial cells (HUVECs) activated by lipopolysaccharide and leukocyte-endothelial cell adhesion.

METHODSThe effects of FA on E-selectin and E-selectin mRNA expression were determined by flow cytometry and reverse transcription polymerase chain reaction. The effect of FA on HL60-HUVEC adhesion was evaluated with the method of staining the cells by Rose Bengal.

RESULTSThe expression of E-selectin and E-selectin mRNA were down regulated by FA (0.62 and 0.41 mmol x L(-1), respectively). HL60 cells adhered to activated HUVECs were also reduced by FA (0.62 and 0.41 mmol x L(-1), respectively).

CONCLUSIONFA can inhibit the expression of E-selectin and E-selectin mRNA and HL60-HUVEC adhesion. This may contribute to its protective effect against ischemia-reperfusion injury.

Cell Adhesion ; drug effects ; Cells, Cultured ; Coumaric Acids ; pharmacology ; E-Selectin ; biosynthesis ; genetics ; Endothelial Cells ; metabolism ; HL-60 Cells ; physiology ; Humans ; RNA, Messenger ; biosynthesis ; genetics ; Umbilical Veins ; cytology

OBJECTIVETo develop a monoclonal antibody (mAb) directed to FVIII C2 domain and investigate its effect on FVIII activity.

METHODSFVIII C2 protein was expressed in E. coli and purified. A murine antihuman FVIII C2 domain mAb SZ-132 was developed by standard hybridoma technology and characterized. In coagulation assays, different concentrations of SZ-132 were incubated with freshly collected pooled human plasma and the residual activity of FVIII and activated partial thromboplastin time (APTT) were determined. The effects of SZ-132 on rhFVIII binding to purified human vWF, phosphatidylserine (PS) and platelets were assessed by enzyme linked immunosorbent assays (ELISA).

RESULTSSZ-132 could inhibit FVIII procoagulant activity in a dose-dependent manner within the concentrations of 0-25 microg/ml and the FVIII activity was completely inhibited on above 25 microg/ml. It could also prevent rhFVIII from binding to vWF, PS and platelets.

CONCLUSIONSSZ-132 is a neutralizing mAb against FVIII C2 domain and can inhibit FVIII procoagulant activity by preventing FVIII from binding to vWF and PS.

Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Antibodies, Neutralizing ; biosynthesis ; immunology ; Factor VIII ; immunology ; metabolism ; Humans ; Male ; Mice ; Mice, Inbred BALB C

BACKGROUNDIt is known that excessive release of glutamate can induce excitotoxicity in neurons and lead to seizure. Dexamethasone has anti-seizure function. The aim of this study was to investigate glutamate-dexamethasone interaction in the pathogenesis of epilepsy, identify differentially expressed genes in the hippocampus of glutamate-induced epileptic rats by mRNA differential display, and observe the effects of dexamethasone on these genes expression.

METHODSSeizure models were established by injecting 5 microl (250 microg/microl) monosodium glutamate (MSG) into the lateral cerebral ventricle in rats. Dexamethasone (5 mg/kg) was injected intraperitoneally at 30 minutes after MSG inducing convulsion. The rats' behavior and electroencephalogram (EEG) were then recorded for 1 hour. The effects of dexamethasone on gene expression were observed in MSG-induced epileptic rats at 1 hour and 6 hours after the onset of seizure by mRNA differential display. The differentially expressed genes were confirmed by Dot blot.

RESULTSEEG and behaviors showed that MSG did induce seizure, and dexamethasone could clearly alleviate the symptom. mRNA differential display showed that MSG increased the expression of some genes in epileptic rats and dexamethasone could downregulate their expression. From more than 10 differentially expressed cDNA fragments, we identified a 226 bp cDNA fragment that was expressed higher in the hippocampus of epileptic rats than that in the control group. Its expression was reduced after the administration of dexamethasone. Sequence analysis and protein alignment showed that the predicted amino acid sequence of this cDNA fragment kept 43% identity to agmatinase, a member of the ureohydrolase superfamily.

CONCLUSIONSThe results of the current study suggest that the product of the 226 bp cDNA has a function similar to agmatinase. Dexamethasone might relax alleviate seizure by inhibiting expression of the gene.

Animals ; Base Sequence ; Dexamethasone ; pharmacology ; Electroencephalography ; drug effects ; Epilepsy ; chemically induced ; drug therapy ; genetics ; Gene Expression Profiling ; Gene Expression Regulation ; drug effects ; Male ; Molecular Sequence Data ; Oligonucleotide Array Sequence Analysis ; Rats ; Rats, Sprague-Dawley ; Sodium Glutamate ; pharmacology

OBJECTIVETo prepare anti-von Willebrand factor A3 (vWF-A3) domain monoclonal antibodies(mAbs) which block vWF-A3 binding to collagen, and characterize their biochemical properties and functions.

METHODSBALB/c mice were immunized with purified recombinant vWF-A3 protein (rvWF-A3). Murine anti-human vWF-A3 mAbs were developed by standard hybridoma technology and identified with ELISA. The recognition of the mAbs with rvWF -A3 and reduced human vWF was identified by Western-blot. The effect of mAbs on binding of purified human vWF to human placenta or calf skin collagen III was studied with collagen binding inhibition test.

RESULTSA group of 30 murine anti-human vWF-A3 mAbs was obtained, from which 2 clones were identified as inhibitory ones and designated as SZ-123 and SZ-125. SZ-123 and SZ-125 could react specifically with human vWF and rvWF-A3 respectively, while neither of them reacted with rvWF-A1 and rvWF-A2. Western-blot showed that SZ-123 and SZ-125 could recognize a 27 x 10(3) band of rvWF-A3 and 2 reduced human vWF bands at 250 x 10(3) and 170 x 10(3). SZ-123 and SZ-125 not only inhibited the binding of purified human vWF (1.5 and 3.0 microg/ml) to human type III collagen and to calf skin collagen III in a dose dependent manner, but also inhibited the binding of plasma vWF from human, rhesus monkeys or Beagle dogs to the two collagens.

CONCLUSIONSZ-123 and SZ-125 are neutralizing mAbs against vWF-A3 domain and may have therapeutic potential as an antithrombotic agent.

Animals ; Antibodies, Monoclonal ; immunology ; isolation & purification ; Collagen ; immunology ; Mice ; Mice, Inbred BALB C ; von Willebrand Factor ; immunology

This study was aimed to investigate the expressions of vascular endothelial growth factor (VEGF) and cyclooxygenase-2 (COX-2) in patients with multiple myeloma (MM) and its clinical significance. Expression of VEGF was detected by enzyme linked immunosorbent assay (ELISA) and the level of COX-2 was detected by Western blot. The results showed that the serum VEGF level of multiple myeloma patients (365.34 +/- 65.63 pg/ml) was higher than that in the normal persons (122.52 +/- 39.29 pg/ml) (p < 0.05); the serum VEGF level of patients at advanced stage (395.07 +/- 54.90) pg/ml was higher than those at stable stage (300.33 +/- 44.22) pg/ml (p < 0.05). The serum Cox-2 positive rate in the patients (31%) was higher than that in normal persons (0%) (p < 0.01); the serum Cox-2 positive rate in the patients at advanced stage (50%) was higher than those at stable stage (21%) (p < 0.01). It is concluded that VEGF and COX-2 may play an important role in the pathogenesis and development of multiple myeloma, they can be used to evaluate the status of patients with MM.
Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Cyclooxygenase 2 ; blood ; Humans ; Middle Aged ; Multiple Myeloma ; blood ; Vascular Endothelial Growth Factor A ; blood

OBJECTIVETo identify a mutation G2113-->A in the glycoprotein (GP)IX gene associated with Bernard-Soulier syndrome (BSS) and to investigate BSS pathogenesis.

METHODSAllele-specific restriction enzyme was used to analyze the samples of patient, her mother, her brother and 40 healthy volunteers. Site-directed mutagenesis was performed to construct a expression vector PD-IXG2113A harboring the mutation G2113-->A. Chinese hamster ovary (CHO) cells were transiently cotransfected with plasmids harboring the entire coding region of GPIbalpha, GPIbeta and GPIX or mutant GPIX, respectively. Expression of GPIbalpha and GPIX in transfected CHO cells were analysed with flow cytometer. GPIbalpha and GPIX in the cytoplasma of transfected CHO cells were analysed by immunostaining and Western blotting.

RESULTSThe patient was found to be homozygosity of the substitution, her mother and her brother be heterozygous. Expressions of GPIbalpha and GPIX in mutant CHO cells were remarkably reduced, but abundant in the cytoplasma.

CONCLUSIONThe mutation of Ala139(GCC)-->Thr(ACC) in the GPIX did not affect synthesis and assembly of GPIb/IX complex but influence its anchoring and expression on the cell surface, which was responsible for BSS.

Adult ; Animals ; Bernard-Soulier Syndrome ; genetics ; Blotting, Western ; CHO Cells ; Cricetinae ; Female ; Humans ; Mutation ; Platelet Glycoprotein GPIb-IX Complex ; genetics