The leucoanthocyantin reducase (LAR) gene, an important functional gene of catechins biosynthesis pathway, was cloned from Fagopyrum dibotrys (D.Don) Hara by degenerate PCR and rapid amplification of cDNA ends (RACE). The full-length cDNA of FdLAR is 1 581 bp (GenBank accession: JN793953), containing a 1 176 bp ORF encoding a 391 amino acids protein, and its 3'-untranslated region has an obvious polyadenylation signal. The recombinant plasmid containing FdLAR completed ORF was transformed into E. coli BL21 (DE3). The target fusion peptide with molecular weight of 66 kD was expressed under the condition of 16 degrees C and induced by IPTG at final concentration of 1.0 mmol x L(-1). Bioinformation analysis indicated that the amino acid sequence of FdLAR showed great homology to other LAR with the NADB-Rossmann conversed domain in the N-terminus. Real-time quantitative PCR was used to detect the expression levels of FdLAR gene during different development periods. The determination of flavonoids contents in appropriate rhizomes showed that the relationship between FdLAR gene expression and the accumulation of flavonoids displayed different trends during vegetative growth and reproductive growth stages, suggesting that the FdLAR gene may be involved in the pathway of flavonoid metabolisms in Fagopyrum dibotrys.

Objective:To evaluate the physiologic work of breathing (WOBphy)as parameter of respiratory weaning and extubation. Method: Patient work of breathing (WOBt) and imposed work of breathing (WOBimp) were measured. WOBphy was obtained with WOBt minuting WOBimp. In the patients who did not meet conventional respiratory weaning parameters ,if WOBphy

OBJECTIVETo investigate the relationship between the excitotoxicity and serum-inducible kinase (SNK) and spine-associated Rap GTPase-activating protein (SPAR) pathway in primary hippocampal neuron injury induced by glutamate and furthermore, to explore the molecular mechanism of neuroprotection of Zibu Piyin Recipe (ZBPYR) and the relationship between ZBPYR and the morphological regulation of dendritic spines.

METHODSThe serum containing ZBPYR was prepared by seropharmacology. Reverse transcription and polymerase chain reaction (RT-PCR) was used to detect the expression of mRNA for SNK, SPAR, postsynaptic density protein 95 (PSD-95) and N-methyl-D-aspartate (NMDA) receptor subunits (NR1, NR2A and NR2B) in primary rat hippocampal neuron cultures after pretreatment with 10 micromol/L glutamate and ZBPYR serum.

RESULTSZBPYR serum pretreatment resulted in a significant down-regulation of glutamate-induced SNK mRNA expression (P<0.05). Significant up-regulation was seen on the mRNA expression of SPAR and PSD-95 (P<0.05). All these changes were dose-dependent. The mRNA expression of NR1, NR2A and NR2B was down-regulated to different degrees (P<0.05).

CONCLUSIONThe mechanism of effect of ZBPYR on glutamate-induced excitotoxicity may be related to the regulation of SNK-SPAR signal pathway. ZBPYR may play a role in protecting and maintaining the normal morphology and structure of dendritic spines, which may be achieved by inhibiting the excessive activation of NMDA receptors.

Animals ; Cells, Cultured ; Disks Large Homolog 4 Protein ; Drugs, Chinese Herbal ; pharmacology ; GTPase-Activating Proteins ; genetics ; metabolism ; Gene Expression Regulation ; drug effects ; Glutamic Acid ; toxicity ; Hippocampus ; drug effects ; enzymology ; pathology ; Intracellular Signaling Peptides and Proteins ; genetics ; metabolism ; Membrane Proteins ; genetics ; metabolism ; Neurons ; drug effects ; enzymology ; pathology ; Protein Kinases ; genetics ; metabolism ; Protein-Serine-Threonine Kinases ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Serum

Objective To study the genotypes of 102 strains of methicillin-resistant Staphylococcus aureus(MRSA)collected consecutively in 2002 in our hospital Method Multiplex PCR was used to genotype Staphylococcal chromosomal cassette mec(SCCmec)element and its variants.Results Among 102 strains of MRSA,the genotypes were as follows:SCCmec-Ⅲ(94 strains),SCCmec-ⅢA(4 strains), SCCmec-Ⅳ(2 stains),SCCmec-Ⅰ(2 stains).Conclusion The predominant genotype of MRSA circulating in this hospital in 2002 was SCCmec-Ⅲ by multiplex PCR.

BACKGROUNDWall shear stress is an important factor in the destabilization of atherosclerotic plaques. The purpose of this study was to assess the distribution of wall shear stress in advanced carotid plaques using high resolution magnetic resonance imaging and computational fluid dynamics.

METHODSEight diseased internal carotid arteries in seven patients were evaluated. High resolution magnetic resonance imaging was used to visualize the plaque structures, and the mechanic stress in the plaque was obtained by combining vascular imaging post-processing with computational fluid dynamics.

RESULTSWall shear stresses in the plaques in all cases were higher than those in control group. Maximal shear stresses in the plaques were observed at the top of plaque hills, as well as the shoulders of the plaques. Among them, the maximal shear stress in the ruptured plaque was observed in the rupture location in three cases and at the shoulder of fibrous cap in two cases. The maximal shear stress was also seen at the region of calcification, in thrombus region and in the thickest region of plaque in the other three cases, respectively.

CONCLUSIONDetermination of maximal shear stress at the plaque may be useful for predicting the rupture location of the plaque and may play an important role in assessing plaque vulnerability.

Aged ; Carotid Arteries ; pathology ; physiopathology ; Carotid Artery Diseases ; pathology ; physiopathology ; Computer Simulation ; Female ; Humans ; Magnetic Resonance Imaging ; methods ; Male ; Middle Aged ; Stress, Mechanical

This study was purposed to detect the balance and the activity change of cytotoxic T cell subsets in aplastic anemia (AA) patients, myelodysplastic syndrome (MDS) patients and acute myeloid leukemia (AML) patients, and to explore the cellular immune mechanism for abnormal hematopoiesis of the three diseases, so as to provide experimental basis for the choice of clinical treatment. The proportion of the cytotoxic T cells and part of the T-cells subsets in peripheral blood were detected by flow cytometry in 35 cases of MDS, including 19 refractory anemia (MDS-RA), 16 refractory anemia with excess blasts (MDS-RAEB), 17 AA, 15 AML patients and 10 normal donors respectively. The results showed that compared with the control group, the percentage of Tc1, Tc1/Tc2, CD8(+)HLA-DR(+), CD3(+)CD8(+)CD28(+), CD8(+)CD45RO(+) cells was significantly higher and the percentage of CD8(+)CD45RA(+) was significantly lower in AA and MDS-RA group. There was no difference in the percentage of Tc2 cells between AA/MDS-RA and normal controls; the percentage of CD8(+)CD45RO(+) cells was significantly higher and the percentage of Tc1, CD3(+)CD8(+)CD28(+), CD8(+)HLA-DR(+) was significantly lower in MDS-RAEB group, the percentage of CD8(+)CD45RA(+) was lower but the difference was not significant, and there was no difference in the percentage of Tc, Tc1/Tc2 cells between MDS-RAEB group and the control group. The percentage of Tc2 cells was significantly higher and the percentage of other parameters was significantly lower in AML group than those of normal controls. It is concluded that the cellular immune statuses in AA, the different stages of MDS and AML are different. In AA and the early stage of MDS, the balance of Tc1/Tc2 shifts to Tc1, and the activation of T-cell subsets increases. In the late stage of MDS and AML, the balance of Tc1/Tc2 shifts to Tc2, the activation of T-cell subsets decreases. The former may be closely related to bone marrow failure while the latter may be one of the important mechanisms in which the malignant clones escape from immune effect.
Adolescent ; Adult ; Aged ; Anemia, Aplastic ; immunology ; pathology ; CD8-Positive T-Lymphocytes ; cytology ; Case-Control Studies ; Female ; Flow Cytometry ; Humans ; Leukemia, Myeloid, Acute ; immunology ; pathology ; Lymphocyte Count ; Male ; Middle Aged ; Myelodysplastic Syndromes ; immunology ; pathology ; Young Adult

The leucoanthocyantin reducase (LAR) gene, an important functional gene of catechins biosynthesis pathway, was cloned from Fagopyrum dibotrys (D.Don) Hara by degenerate PCR and rapid amplification of cDNA ends (RACE). The full-length cDNA of FdLAR is 1 581 bp (GenBank accession: JN793953), containing a 1 176 bp ORF encoding a 391 amino acids protein, and its 3'-untranslated region has an obvious polyadenylation signal. The recombinant plasmid containing FdLAR completed ORF was transformed into E. coli BL21 (DE3). The target fusion peptide with molecular weight of 66 kD was expressed under the condition of 16 degrees C and induced by IPTG at final concentration of 1.0 mmol x L(-1). Bioinformation analysis indicated that the amino acid sequence of FdLAR showed great homology to other LAR with the NADB-Rossmann conversed domain in the N-terminus. Real-time quantitative PCR was used to detect the expression levels of FdLAR gene during different development periods. The determination of flavonoids contents in appropriate rhizomes showed that the relationship between FdLAR gene expression and the accumulation of flavonoids displayed different trends during vegetative growth and reproductive growth stages, suggesting that the FdLAR gene may be involved in the pathway of flavonoid metabolisms in Fagopyrum dibotrys.
Amino Acid Sequence ; Anthocyanins ; metabolism ; Cloning, Molecular ; Fagopyrum ; enzymology ; genetics ; growth & development ; Flavonoids ; analysis ; Gene Expression Regulation, Enzymologic ; Gene Expression Regulation, Plant ; Genes, Plant ; Molecular Sequence Data ; Oxidoreductases ; genetics ; metabolism ; Phylogeny ; Plant Proteins ; genetics ; metabolism ; Plants, Medicinal ; enzymology ; genetics ; growth & development ; Rhizome ; genetics

OBJECTIVETo evaluate the role of gastric mucosa apoptosis in the stress of ischemic stroke, and to discuss the relationship between gastric mucosa apoptosis and gastric barrier.

METHODSTen dogs were artificially made ischemic stroke by operation (IS group), and another 10 shamly-operated dogs were served as control group. Sucrose permeability were measured after the operation. All dogs were sacrificed 24 hours after operation to measure the gastric mucosal apoptosis index, gastric gross classification, and histological score.

RESULTSThe gastric mucosal apoptosis index in the IS group were significantly higher than in the control group (14.83 +/- 4.41 vs. 5.60 +/- 2.61, P < 0.05). The gastric mucosal apoptosis index were correlated with the sucrose permeability (r = 0. 89, P < 0.05) , gastric gross classification (r = 0. 87, P < 0.05), and histological score (r = 0.92, P < 0.05).

CONCLUSIONSAlthough ischemic stroke will not cause the obvious damage in the respiratory and circulatory system, it is responsible for the apoptosis of epithelial cell in the gastric mucosa and gastric barrier dysfunction. The apoptosis index is closely correlated with the damage of the function and morphology of the gastric barrier, indicating that the epithelial cell apoptosis acceleration in the gastric mucosa may result in the damage of gastric barrier function.

Animals ; Apoptosis ; physiology ; Dogs ; Epithelial Cells ; pathology ; Gastric Mucosa ; pathology ; In Situ Nick-End Labeling ; Random Allocation ; Stroke ; pathology ; physiopathology

Objective To determine the safety and efficacy of intra-arterial urokinase in the treatment of acute cerebral infarction (ACI) patients with computed tomography perfusion-based selection within a 6-9 h window.Methods Fifty-two ACI patients,with computed tomography perfusion imaging (CTPI) identifying thresholds for salvageable penumbra,were randomly assigned to intra-arterial thrombolysis with urokinase (group A) and conventional anti-platelet aggregation (group B) within a 6-9 h window.Whole brain digital subtraction angiography (DSA) was done at pre-and post-treatment to observe the recanalization of occlusive vessels in group A.The National Institutes of Health Stroke scale (NIHSS) 24 h and 7 d after treatment,and modified Rankin Scale (mRS) and Barthel Index (BI) 90 d after treatment were used to evaluate the efficacy.Results In group A,15 patients showed successful recanalization (thrombolysis in myocardial infarction [TIMI] index:grade Ⅲ in 9 and grade Ⅱ in 6) and 12 patients showed unsuccessful recanalization (TIMI index:grade Ⅰ in 6 and grade 0 in 6) with a successful recanalization rate of 55.56％.More obvious NIHSS improvement 24 h and 7 d after treatment in group A was observed than that in group B (P＜0.05),and more patients with favorable outcomes based on mRS and BI in group A were noted than those in group B (P＜0.05).In addition,the incidence of cerebral hemorrhage within 24 h of treatment between the two groups was similar (P＞ 0.05).Conclusions Intra-arterial thrombolysis with urokinase is safe and effective for ACI patients within a 6-9 hour window under the guidance of CTPI.

Tweety-homolog 1 (Ttyh1) is expressed in neural tissue and has been implicated in the generation of several brain diseases. However, its functional significance in pain processing is not understood. By disrupting the gene encoding Ttyh1, we found a loss of Ttyh1 in nociceptors and their central terminals in Ttyh1-deficient mice, along with a reduction in nociceptor excitability and synaptic transmission at identified synapses between nociceptors and spinal neurons projecting to the periaqueductal grey (PAG) in the basal state. More importantly, the peripheral inflammation-evoked nociceptor hyperexcitability and spinal synaptic potentiation recorded in spinal-PAG projection neurons were compromised in Ttyh1-deficient mice. Analysis of the paired-pulse ratio and miniature excitatory postsynaptic currents indicated a role of presynaptic Ttyh1 from spinal nociceptor terminals in the regulation of neurotransmitter release. Interfering with Ttyh1 specifically in nociceptors produces a comparable pain relief. Thus, in this study we demonstrated that Ttyh1 is a critical determinant of acute nociception and pain sensitization caused by peripheral inflammation.