Background Fractalkine is involved in the impairment of endothelium by mediating inflammatory cell chemotaxis.Aspirin inhibites many kinds of cytokine expression.Objective To investigate the effect of aspirin on tumor necrosis factor ?(TNF-?) stimulated fractalkine expression in human umbilical vein endothelial cells (HUVEC) and its mechanism.Methods HUVEC were grouped as follows:control group;TNF-?-stimu- lated;TNF-?+NS-398;TNF-?+PDTC and TNF plus various concentration of aspirin (0.02,0.2,1.5 mmol/L). The level of mRNA and protein expression of fractalkine and nuclear factor(NF)-kB p65 was determined by RT- PCR and Western blot.Results 1)Fractalkine mRNA and protein level was increased after 4 ?g/L TNF-? stim- ulation in HUVEC(both P

Objective To study the expression and the relation of vascular endothelial growth factor (VEGF)and matrix metal proteinase-2(MMP-2)in rectal cancer and evaluate their roles in rectal carcinogen- esis and development.Methods The expression of VEGF and MMP-2 in 52 cases of rectal cancer was de- tected by immunohistochemical SP technique.12 cases normal rectal tissue served as the control group.Re- suits The expression of VEGF in rectal carcinoma(67.3 %)was much higher than that in control group(P

OBJECTIVETo investigate the expression of cytokeratins (CK) in the normal human gingival epithelium and to explore the difference between junctional epithelium (JE), oral epithelium (OE) and sulcular epithelium (SE).

METHODSTeeth specimens with gingival tissue were collected from 5 people. Paraffin-embedded sections were stained with monoclonal antibodies responded respectively to human CK5/6, 7, 8/18, 10/13, 16, 17, 19, 20.

RESULTSCK7 and 17 was not expressed in all strata of JE, OE and SE. CK5/6 and 20 were weekly or moderately expressed in the suprabasal, and not expressed in the basal layer of all three epithelia. CK10/13 and 16 were positive in all strata of JE and in the suprabasal layers of OE and SE. CK10/13 was moderately to strongly expressed and CK16 was weekly to moderately expressed. The staining for CK19 was intense in all strata of JE and the basal layer of OE and SE. There was a remarkable demarcation between JE and SE. The pattern of CK8/18 expression was similar to that of CK19, but was weaker. Besides the basal layer, some suprabasal layers close to the basal layer were stained.

CONCLUSIONSJE is an unique non-differentiated stratified epithelium different from OE and SE. CK19 would be a histological marker and CK10/13, 16 would be the cellular markers to differentiate JE from OE and SE.

Epithelial Attachment ; cytology ; metabolism ; Gingiva ; metabolism ; Humans ; Keratins ; metabolism ; Mouth Mucosa ; metabolism

2600 Anthers from T0 modified cry1 Ac-transgenic rice lines of Minghui 81, an elite restoring line of commercial CMS indica hybrid rice, were cultured on SK3 media. 83 green plantlets were recovered, 43 double haploid (DH) and 40 haploid among them. Results of PCR analyzes indicated that 55 plants of 83 were harbored the cry1Ac gene, and the ratio of cry1Ac-positive against cry1Ac-negative was 2:1 (55/28). 36 putative transgenic DH plants were further confirmed by Southern blot. ELISA detection showed that Cry1Ac level in different transgenic rice plants of the same cry1Ac-DH clone was almost equal and the highest one amount to 0.25% of the total soluble protein. Pest insect-resistant bioassay at field trials demonstrated that some of the homozygous cry1Ac-transgenic rice plants not only showed high-level resistance against striped stem borer (Chilo suppressalis) but also retained elite agronomy characters. These results demonstrated that rice anther culture has a great value in rice molecular breeding.
Animals ; Bacillus thuringiensis ; genetics ; Bacterial Proteins ; genetics ; metabolism ; Bacterial Toxins ; Blotting, Southern ; Culture Techniques ; DNA, Plant ; genetics ; Endotoxins ; genetics ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Gene Expression ; Hemolysin Proteins ; Immunity, Innate ; genetics ; Moths ; growth & development ; Oryza ; genetics ; growth & development ; parasitology ; Plant Structures ; genetics ; growth & development ; parasitology ; Plants, Genetically Modified

OBJECTIVETo study diffuse alveolar hemorrhage (DAH) in patients with hematologic malignancies after chemotherapy and discuss the possible etiology and appropriate therapy.

METHODSSymptoms, physical examinations, laboratory examination, chest radiographs or computed tomographic (CT) scans, treatments and outcomes of two patients with acute lymphoblastic leukemia (ALL) and non-Hodgkin lymphoma (NHL) each after chemotherapy were presented.

RESULTSBoth of the patients developed cough, progressive dyspnea, a drop of hemoglobin level, hypoxemia and widespread pulmonary infiltrate on chest radiographs or CT scans after chemotherapy. Moreover, case 1 (ALL) had high fever and bloody fluid drained from the intubation of mechanical ventilation, case 2 (NHL) developed continual hemoptysis. They were diagnosed as DAH and improved significantly after intermediate- or high-dose corticosteroid therapy.

CONCLUSIONSDAH is a rare fatal acute noninfectious pulmonary complication in patients with hematologic malignancies after chemotherapy. Early accurate diagnosis, identifying the underlying cause and appropriate treatment are critical for the management of DAH.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; Female ; Hemorrhage ; drug therapy ; etiology ; Humans ; Lymphoma, Non-Hodgkin ; drug therapy ; Male ; Methylprednisolone ; therapeutic use ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; Pulmonary Alveoli

OBJECTIVETo observe the blood enriching function of Siwu Tang and its effect on Epo and G-CSF gene expressions in bone marrow of blood deficiency mice, and thus provide the basis for understanding the molecular mechanism of blood enriching function of Siwu Tang.

METHODThe animal model of blood deficiency were established in the mice by using 3.5 Gy60Co gamma-ray. Peripheral blood cells were analyzed and CFU-GM, BFU-E, CFU-E and CFU-mix were counted in bone marrow colony cultured. Both Epo and G-CSF gene expressions in bone marrow were measured with RT-PCR.

RESULTSiwu Tang significantly increased the number of peripheral blood cells and the amount of CFU-GM, BFU-E, CFU-E and CFU-mix in bone marrow and enhanced Epo and G-CSF gene expression in bone marrow in the mice with blood deficiency.

CONCLUSIONThe promotion of Epo and G-CSF gene expressions in bone marrow may be one of the mechanisms underlying the blood enriching function of Siwu Tang decoction.

Animals ; Bone Marrow ; metabolism ; Bone Marrow Cells ; cytology ; Cell Count ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Erythroid Precursor Cells ; cytology ; Erythropoietin ; biosynthesis ; genetics ; Female ; Granulocyte Colony-Stimulating Factor ; biosynthesis ; genetics ; Leukocyte Count ; Medicine, Chinese Traditional ; Mice ; Mice, Inbred C57BL ; Plants, Medicinal ; chemistry ; RNA, Messenger ; biosynthesis ; genetics ; Whole-Body Irradiation

BACKGROUNDFractalkine is an important chemokine mediating local monocyte accumulation and inflammatory reactions in the vascular wall. Aspirin inhibits inflammatory cytokine expression closely related to atherosclerosis through the way independent of platelet and cyclooxygenase (COX). There has been no report about the effect of aspirin on fractalkine expression. We aimed to determine the fractalkine expression in human umbilical vein endothelial cell (HUVEC) stimulated by tumor necrosis factor (TNF)-alpha and the effect of aspirin intervention.

METHODSSix of 8 HUVEC groups received either different concentrations of aspirin (0.02, 0.2, 1.0, 5.0 mmol/L) or 40 micromol/L pyrrolidinecarbodithioc acid (PDTC) or 0.5 micromol/L NS-398. The other two groups were negative control and positive control (TNF-alpha-stimulated). After being incubated for 24 hours, cells of the 8 groups except the negative control one were stimulated with TNF-alpha (4 ng/ml) for another 24 hours. After that, the cells were collected for RNA isolation and protein extraction.

RESULTSBoth mRNA and protein expressions of fractalkine in HUVEC were upregulated by 4 ng/ml TNF-alpha stimulation. Aspirin inhibited fractalkine expression in a dose-dependent manner at mRNA and protein levels. Nuclear factor-kappa B inhibitor, PDTC, effectively decreased the fractalkine expression. Fractalkine expression was not influenced by COX-2 selective inhibitor NS-398. COX-1 protein expression was not changed by either TNF-alpha stimulation or aspirin, PDTC, NS-398 intervention. Both mRNA and protein expression of COX-2 in HUVEC were upregulated by 4 ng/ml TNF-alpha stimulation. Aspirin decreased COX-2 expression in a dose-dependent manner at mRNA and protein levels.

CONCLUSIONSTNF-alpha-stimulated fractalkine expression is suppressed by aspirin in a dose-dependent manner through the nuclear factor-kappa B p65 pathway.

Aspirin ; pharmacology ; Blotting, Western ; Cell Line ; Chemokine CX3CL1 ; genetics ; metabolism ; Endothelial Cells ; drug effects ; metabolism ; Gene Expression Regulation ; drug effects ; Humans ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Necrosis Factor-alpha ; pharmacology ; Umbilical Veins ; cytology

BACKGROUNDBrain-derived neurotrophic factor (BDNF) is a stress-responsive intercellular messenger modifying hypothalamic-pituitary-adrenal (HPA) axis activity. The interaction between stress and age in BDNF expression is currently not fully understood. This study was conducted to observe unpredictable stress effect on behavior and BDNF expression in CA3 subfield (CA3) and dentate gyrus of hippocampus in different aged rats.

METHODSForty-eight Wistar rats of two different ages (2 months and 15 months) were randomly assigned to six groups: two control groups and four stress groups. The rats in the stress group received three weeks of unpredictable mild stress. The depression state and the stress level of the animals were determined by sucrose preference test and observation of exploratory behavior in an open field (OF) test. The expressions of BDNF in CA3 and dentate gyrus of the hippocampus were measured using immunohistochemistry.

RESULTSAge and stress had different effects on the behavior of different aged animals (age: F = 6.173, P < 0.05, stress: F = 6.056, P < 0.05). Stress was the main factor affecting sucrose preference (F = 123.608, P < 0.05). Decreased sucrose preference and suppressed behavior emerged directly following stress, lasting to at least the eighth day after stress in young animals (P < 0.05). The older stress rats showed a lower sucrose preference than young stress rats (P < 0.05). Older control rats behaved differently from the younger control animals in the OF test, spending more time in the central square (P < 0.05), exhibiting fewer vertical movements (P < 0.05) and less grooming (P < 0.05). Following exposure to stress, older-aged rats showed no obvious changes in vertical movement and grooming. This indicates that aged rats were in an unexcited state before the stress period, and responded less to stressful stimuli than younger rats. There was significantly lower BDNF expression in the CA3 and dentate gyrus regions of the hippocampus following stress in both age groups (P < 0.05), a reduction that was still present at the eighth day after stress (P < 0.05). Stress and age were the main factors affecting the expression of BDNF (F = 9.408, P < 0.05; F = 106.303, P < 0.05). The aged stress group showed lower BDNF expression compared to the young stressed group at every testing time point.

CONCLUSIONStress has age-dependent effects on behavioral responses and hippocampal BDNF expression in rats.

Age Factors ; Animals ; Behavior, Animal ; Brain-Derived Neurotrophic Factor ; analysis ; genetics ; Chronic Disease ; Dentate Gyrus ; chemistry ; Exploratory Behavior ; Hippocampus ; chemistry ; Immunohistochemistry ; Rats ; Rats, Wistar ; Stress, Psychological ; metabolism ; psychology ; Sucrose ; administration & dosage

To study the effect of Siwu decoction on the function and expression of P-glycoprotein (P-gp) in Caco-2 cells. The Real-time quantitative poly-merase chain reaction (Q-PCR) was used to analyze the mRNA expression of MDR1 gene in Caco-2 cells. Flow cytometer was used to study the effect of Siwu decoction on the uptake of Rhodamine 123 in Caco-2 cells, in order to evaluate the efflux function of P-gp. Western blotting method was used to detect the effect of Siwu decoction on the P-gp protein expression of Caco-2 cells. Compared with the blank control group, after Caco-2 incubation with Siwu decoction at concentrations of 3.3, 5.0, 10.0 g x L(-1) for 24, 48, 72 h, the mRNA expression of MDR1 was up-regulated, suggesting the effect of Siwu decoction in inducing the expression of MDR1. After the administration with Siwu decoction in Caco-2 cells for 48 h, the uptake of Rhodamine 123 in Caco-2 cells decreased by respectively 16.6%, 22.1% (P < 0.05) and 45.4% (P < 0.01), indicating that the long-term administration of Siwu decoction can enhance the P-gp efflux function of Caco-2 cells. After the incubation of Caco-2 cells with Siwu decoction for 48 h, the P-gp protein expression on Caco-2 cell emebranes, demonstrating the effect of Siwu decoction in inducing the protein expression of P-gp.
ATP Binding Cassette Transporter, Sub-Family B ; genetics ; metabolism ; ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Caco-2 Cells ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Up-Regulation ; drug effects

OBJECTIVETo investigate the influence of ethylbenzene on oxidative damage, ultrastructure and the expressions of apoptosis-related genes in the rat brain tissues.

METHODSFour groups of 10 males of Sprague-Dawley rats were allocated randomly, and inhaled daily with different doses of ethylbenzene: 0, 433.5 mg/m³, 4335.0 mg/m³, and 6500.0 mg/m³ 6 h daily, 5 days per week for 13 weeks. The contents of glutathione (GSH) and malondialdehyde (MDA) and activity of acetylcholinesterase (AChE) were assayed, respectively. The ultrastructure of brain tissues was observed via electron microscope. The gene expression levels of Bax, Bcl-2, cytochrome C, caspase-9 and caspase-3 in brain tissues were measured by real-time polymerase chain reaction (PCR), respectively.

RESULTSThe contents of MDA [(2.03 ± 0.56), (4.17 ± 1.31) nmol/mg pro] in the brain tissues of 4335.0 mg/m³ and 6500.0 mg/m³ ethylbenzene-treated groups were significantly higher than that [(1.08 ± 0.26) nmol/mg pro] in the control group (P < 0.05), while AChE activities [(0.321 ± 0.066), (0.276 ± 0.031), (0.202 ± 0.041) U/mg] and GSH contents [(35.19 ± 15.08), (33.42 ± 15.32), (27.99 ± 7.53) mg/g pro] in all ethylbenzene-treated groups were remarkably depressed (P < 0.05, P < 0.05, respectively). After 6500.0 mg/m³ ethylbenzene inhalation, the nucleolus exhibit demilune with decreased mitochondria. Electrondense of myelin occurred in the injured nerve, ascribing to lipid peroxidationed membrane. The gene expression level of Bax in brain tissue of 4335.0 mg/m³ and 6500.0 mg/m³ ethylbenzene-treated group was significantly higher than that in the control group (P < 0.05). Compared with the control group, the gene expression levels of cytochrome C, caspase-9 and caspase-3 in all ethylbenzene-treated groups were enhanced (P < 0.05, P < 0.05, respectively), while bcl-2 gene expression levels in all ethylbenzene-treated groups were decreased (P < 0.05).

CONCLUSIONEthylbenzene can induce oxidative damage and apoptosis in brain tissues. The apoptotic mechanism might be involved with up-regulation of Bax, cytochrome C, caspase-9 and caspase-3, as well as restraint of Bcl-2.

Animals ; Apoptosis ; Benzene Derivatives ; toxicity ; Brain ; drug effects ; metabolism ; ultrastructure ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Cytochromes c ; metabolism ; DNA Damage ; Female ; Gene Expression ; Male ; Oxidative Stress ; drug effects ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Sprague-Dawley ; bcl-2-Associated X Protein ; metabolism