Asthma ; prevention & control ; Child ; Humans ; Hypersensitivity ; prevention & control

Ductus Arteriosus ; abnormalities ; Fatal Outcome ; Female ; Heart Septal Defects, Atrial ; etiology ; Humans ; Infant, Newborn ; Rubella Syndrome, Congenital ; complications

Objective To observe the expression of growth factors (vascular endothelial growth factor [VEGF],stromal cell-derived factor-1 [SDF-1 ],basic fibroblast growth factor [bFGF],insulin-like growth factor [IGF-1],transforming growth factor-β [TGF-β],platelet-derived growth factor [PDGF],brain derived neurotrophic factor [BDNF],glial cell line-derived neurotrophic factor [GDNF] and nerve growth factor [NGF]) in rat ischemic brain tissues after intravenous implantation of bone marrow stromal cells (BMSCs) and/or endothelial progenitor cells (EPCs). Methods Healthy adult Wistar rats were randomly divided into 4 groups:vehicle group,BMSCs transplantation group,EPCs transplantation group and BMSCs combined with EPCs transplantation group (n=20). The rats were subjected to middle cerebral artery occlusion (MCAO),and 24 h after that,they were intravenously transplanted with either 3×106 BMSCs,EPCs,BMSCs/EPCs or 1 mL physiological saline.Seven d after transplantation,real time-PCR and Western blotting were employed to detect the expressions of VEGF,SDF-1,bFGF,IGF-1,TGF-β,PDGF-BB,BDNF,GDNF and NGF. Results The mRNA expressions of bFGF,VEGF and BNDF in the BMSCs/EPCs transplantation group were significantly higher as compared with those in the other groups (P＜0.05).BMSCs transplantation group enjoyed the highest mRNA levels of NGF,GDNF and TGF-β among all the groups, significantly higher as compared with those in the other groups (P＜0.05),followed by BMSCs/EPCs transplantation group.EPCs transplantation group enjoyed the highest mRNA levels of PDGF,IGF-1 and SDF-1,significantly higher as compared with those in the other groups (P＜ 0.05), followed by BMSCs/EPCs transplantation group. Conclusion BMSCs combined with EPCs implantation can promote the functional rehabilitation in rats after focal cerebral ischemia, which provides new way for improving the transplantation success rate.

OBJECTIVETo investigate the changes in plasma levels of thrombomodulin (TM) and D-dimer (DD) in children with different types of Mycoplasma pneumoniae pneumonia (MPP), and their role in the pathogenesis of MPP in children.

METHODSFifty-two children with MMP were divided into lobar pneumonia (n=30) and interstitial pneumonia groups (n=22) and another 30 healthy children were selected as the control group. Plasma levels of TM and D-D were measured using enzyme-linked immunosorbent assay and latex-enhanced immunoturbidimetric assay, respectively.

RESULTSThe lobar pneumonia, interstitial pneumonia and control groups had median plasma TM levels of 23.83, 15.56 and 8.78 μg/L respectively, with significant differences between the three groups (P<0.01). The lobar pneumonia and interstitial pneumonia groups had significantly higher plasma TM levels than the control group (P<0.01), and the lobar pneumonia group had a significantly higher plasma TM level than the interstitial pneumonia group (P<0.05). Median plasma D-D levels in the lobar pneumonia and interstitial pneumonia groups were significantly higher than the reference value (P<0.01). The lobar pneumonia group had a significantly higher plasma D-D level than the interstitial pneumonia group (0.35 μg/mL vs 0.13 μg/mL; P<0.01), and the percentage of patients with elevated plasma D-D levels was significantly higher in the lobar pneumonia group than in the interstitial pneumonia group (87% vs 59%; P<0.05).

CONCLUSIONSChildren with MPP, especially those with lobar pneumonia, have increased plasma levels of TM and D-D. This suggests that damage to vascular endothelial cells and blood hypercoagulability may be involved in the pathogenesis of MPP.

Adolescent ; Child ; Child, Preschool ; Female ; Fibrin Fibrinogen Degradation Products ; analysis ; Humans ; Male ; Pneumonia, Mycoplasma ; blood ; Protein Multimerization ; Thrombomodulin ; blood

AIMTo develop a capillary gas chromatographic method for the determination and pharmacokinetic study of patchouli alcohol in rat plasma after iv administration.

METHODSThe drug was extracted with ethyl acetate. Eugenol was used as internal standard. The separation was carried out on a HP-5MS quartz capillary column, with high-purity nitrogen as carrier gas and flame ionization detector (FID) as detector. The column temperature was maintained at 80 degrees C for 1 min and then programmed to 200 degrees C at a rate of 15 degrees C x min(-1); it was held at 200 degrees C for 1 min, and then programmed to 290 degrees C at a rate of 60 degrees C x min(-1); the final temperature was held for 1 min. The temperature of both injector and detector was set at 290 degrees C.

RESULTSThe standard curve was linear from 25 to 5 000 microg x L(-1) in rat plasma. The recovery of this method was from 90.0% to 110.0% with satisfactory relative standard deviation (RSD) less than 10.0%. The pharmacokinetic parameters demonstrated patchouli alcohol were consistent with the two-compartment open model and showed linear pharmacokinetics. The T1/2beta, AUC and MRT of patchouli alcohol in patchouli oil were all higher than that of patchouli alcohol.

CONCLUSIONThis method is quick, precise and reliable. The pharmacokinetics of patchouli alcohol is different from that of patchouli alcohol in patchouli oil.

Animals ; Area Under Curve ; Injections, Intravenous ; Lamiaceae ; chemistry ; Male ; Oils, Volatile ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Sesquiterpenes ; blood ; isolation & purification ; pharmacokinetics

OBJECTIVETo investigate the expression of T cell early activation marker (CD(69)) on peripheral CD(4)(+) and CD(8)(+) lymphocytes and serum levels of soluble tumor necrosis factor receptor 1 and 2 (sTNF-R1 and sTNF-R2) in serum and bone marrow in patients with aplastic anemia (AA) and their pathophysiological significance.

METHODSIn vitro activation of T lymphocytes was carried out by whole blood cell culture containing PHA (20 micro g/ml). The CD(69) expressions on CD(4)(+) and CD(8)(+) lymphocytes at 0 h and 4 h after PHA exposure were analyzed by two-color flow cytometry. The levels of sTNF-R1 and sTNF-R2 in serum and bone marrow were measured by ELISA.

RESULTSThe CD(69) expression rates of CD(4)(+) and of CD(8)(+) cells in SAA patients were (8.96 +/- 7.23)% and (10.67 +/- 7.58)%, respectively, and that of CD(8)(+) cells in CAA patients was (7.36 +/- 5.49)% before PHA stimulation. The CD(69) expression rates of CD(4)(+) and of CD(8)(+) cells in SAA patients were (71.73 +/- 11.91)% and (61.74 +/- 13.44)% and in CAA (59.35 +/- 10.15)% and (48.78 +/- 8.25)% respectively, and were significantly elevated after PHA stimulation. CD(69) expression on CD(4)(+) cells was much higher than that on CD(8)(+) cells after stimulation. The levels of the two sTNF-R (sTNF-R1 and sTNF-R2) in peripheral blood and bone marrow of SAA patients were elevated and in the bone marrow of CAA patients were also increased. The serum levels of sTNF-R2 were positively related to the CD(69) expression rates of CD(8)(+) cells before PHA stimulation.

CONCLUSIONIncreased early activation and activated potentials of T lymphocytes, along with abnormally elevated immunologically active molecules might play a major role in the pathogenesis of AA.

Adolescent ; Adult ; Aged ; Anemia, Aplastic ; immunology ; Antigens, CD ; blood ; Antigens, Differentiation, T-Lymphocyte ; blood ; CD4-Positive T-Lymphocytes ; chemistry ; CD8-Positive T-Lymphocytes ; chemistry ; Child ; Female ; Humans ; Lectins, C-Type ; Male ; Middle Aged ; Receptors, Tumor Necrosis Factor ; blood ; Receptors, Tumor Necrosis Factor, Type I ; Receptors, Tumor Necrosis Factor, Type II

OBJECTIVETo investigate the three-dimensional reconstruction methods of the portal vein using 64-slice spiral CT data and the anatomical variation of the portal vein.

METHODSThree-dimensional reconstruction of the portal vein was performed using Mimics software based on the 64-slice spiral CT data of 64 cases. Each model of the portal vein and its branches was evaluated according to the presentation rate, depiction quality and anatomic variation.

RESULTSThe reconstructed model showed a depiction rates of 100% for the 4-grade branches of the portal vein. The stem of the portal vein and the left and right branches of the level III or above were all displayed, but in 2 cases the superior mesenteric vein and in 1 case the spleen vein was displayed only to the level IV. Of the 64 cases, 50 (78.1%) had normal portal vein and 14 (21.9%) showed anatomical variations.

CONCLUSIONThe 3D model vividly mimics the anatomic variations of the portal vein to provide valuable information for surgical plans.

Adult ; Female ; Humans ; Image Processing, Computer-Assisted ; Imaging, Three-Dimensional ; methods ; Male ; Middle Aged ; Portal Vein ; anatomy & histology ; diagnostic imaging ; Tomography, Spiral Computed ; methods ; Young Adult

OBJECTIVETo study the changes in plasma substance P (SP) and calcitonin gene-related peptide (CGRP) levels in children with acute asthma before and after gamma-aminobutyric acid (GABA) treatment.

METHODSSeventy-five children with asthma were randomly assigned to GABA treatment (n=36) and control groups (n=39). Both groups were given conventional treatment for asthma. Besides the conventional treatment, the treatment group was administered with oral GABA (25-30 mg/kg•d). Plasma content of SP and CGRP was measured using ELISA before treatment and after remission.

RESULTSThere were no significant differences in plasma content of SP and CGRP between the GABA treatment and control groups (P＞0.05) before treatment. Plasma content of SP and CGRP in the GABA treatment group was significantly lower than the control group (SP: 57±15 pg/mL vs 127±12 pg/mL; CGRP: 23±10 pg/mL vs 42±8 pg/mL) after remission (P<0.01). Plasma content of SP and CGRP after remission was significantly lower than before treatment (P<0.01) in both groups. There was a significantly positive correlation between plasma SP and CGRP content in asthmatic children (r=0.792, P<0.01).

CONCLUSIONSGABA can significantly decrease plasma levels of SP and CGRP in children suffering from acute asthma.

Asthma ; blood ; drug therapy ; Calcitonin Gene-Related Peptide ; blood ; Child ; Child, Preschool ; Female ; Humans ; Male ; Substance P ; blood ; gamma-Aminobutyric Acid ; pharmacology ; therapeutic use

OBJECTIVETo express and purify the protein coded by the TRAF-type zinc finger domain of myasthenia gravis (MG)-related gene P9 (P9-ZFD) and to prepare P9-ZFD antiserum for detecting expression and subcellular distribution of P9-ZFD protein in the skeletal muscles of patient with MG.

METHODSThe cDNA encoding P9-ZFD was amplified by RT-PCR. The cloned P9-ZFD cDNA was ligated into pET24a, and the P9-ZFD recombinant protein was induced via E. coli. BL21 (DE3) and purified by histidine affinity chromatography. P9-ZFD antiserum was prepared and its titer and specificity were determined by ELISA and Western blot. Expression and subcellular distribution of P9-ZFD protein in the skeletal muscles of MG and control were studied.

RESULTSThe molecular weight of purified P9-ZFD protein was about 30 kD. Its purity was more than 95%. Antiserum specific for P9-ZFD was excellent. P9-ZFD protein is fully confined to the cytoplasm membrane of skeletal muscle cell of MG, obvious immunostaining was absent in the A, I, and Z bands of cytoplasm and no immunoreactivity was observed in the skeletal muscle cell of control.

CONCLUSIONP9-ZFD protein is expressed as a cytoplasm membrane-bound protein and has obvious distribution difference in the skeletal muscle cells of patient with MG and normal control.

Adult ; Cell Membrane ; metabolism ; Escherichia coli ; metabolism ; Female ; Humans ; Muscle Proteins ; biosynthesis ; genetics ; Muscle, Skeletal ; metabolism ; pathology ; Myasthenia Gravis ; metabolism ; Peptide Fragments ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Transfection ; Zinc Fingers

OBJECTIVESTo investigate whether the human PC-3 cell infected with recombinant Ad-PTEN and Ad-p27Kip1 can steadily produce PTEN and p27Kip1 protein and change the biologic behaviors such as cell proliferation, cell cycle and apoptosis. The synergistic effect of PTEN and p27Kip1 on the therapy for prostate cancer has also been investigated.

METHODSWe constructed recombinant adenovirus vector of human tumor suppressor gene PTEN and p27Kip1. The viral titer was examined by plaque assay and the mRNA and protein expressions of PTEN and p27Kip1 in human prostate cancer cell line PC-3 infected with Ad-PTEN and Ad-p27Kip1 were determined by RT-PCR and Western blot respectively. MTT assay was used to determine the effect of PTEN and p27Kip1 on growth and proliferation of PC-3 cell; the change of cell cycle and apoptosis was examined by flow cytometry, and to compare between the combined therapy group and single gene therapy group.

RESULTSThe viral titers of Ad-PTEN and Ad-p27Kip1 were 1.8 x 10(7) pfu/ml and 1.2 x 10(9) pfu/ml respectively. After infected by adenovirus, it had been verified that the mRNA and protein expression of PTEN and p27Kip1 were steady in human PC-3 cell. Ad-PTEN and Ad-p27 Kip1 inhibited the growth and proliferation of PC-3 cells. The progression of cell cycle of PC-3 cell was arrested in G(0)-G(1) phase, meanwhile the apoptosis rate of PC-3 was also affected after Ad-PTEN or/and Ad-p27 Kip1 infected. There was significant difference between combined therapy group and single gene therapy group.

CONCLUSIONThe recombinant Ad-PTEN and Ad-p27Kip1 vector were constructed successfully and the expression of specific PTEN and p27Kip1 was high, steadily in PC-3 cell line. These results suggested that combination of PTEN with p27Kip1 has an application value in treatment of prostate cancer in future.

Adenoviridae ; genetics ; Apoptosis ; drug effects ; Cell Division ; drug effects ; Cell Line, Tumor ; Chromosomes, Human, Pair 10 ; genetics ; Cyclin-Dependent Kinase Inhibitor p27 ; Gene Deletion ; Genetic Therapy ; Genetic Vectors ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; pharmacology ; Male ; PTEN Phosphohydrolase ; genetics ; pharmacology ; Prostatic Neoplasms ; genetics ; physiopathology ; therapy ; Transfection