AIM: To study the effect of platelet-activating factor(PAF) on proliferation of cultured rat airway smooth muscle cells(ASMCs).METHODS: The cells were divided into control group and PAF group. The cells in PAF group were subdivided into four small groups by concentrations of PAF 10 -6 , 10 -7 , 10 -8 , 10 -9 mol?L -1 , MTT assay was used not only to investigate the effects of PAF on proliferation of ASMC but also to confirm the optimal concentration. Flow cytometry and immuneohistochemistry for proliferating cell nuclear antigen (PCNA) were also used to analyse its function on proliferation of ASMC.RESULTS: PAF (10 -6 -10 -9 mol?L -1 ) stimulated the cell proliferation and 10 -7 mol?L -1 PAF reached the maximal effect. The cell percentage of the ASMCs of 107 mol?L -1 PAF subgroup at G_ 0/1 phase (68.67%) was much lower than that of control group (85.57%, P

AIM: To examine whether calcitonin gene-related peptide (CGRP) enhances nitric oxide (NO) level in pulmonary circulation blood and observe the influence of CGRP on mean pulmonary artery pressure (mPAP) in rabbits with acute lung injury (ALI) caused by oleic acid. METHODS: The level of NO was assessed by measuring the presence of nitrite in cervical artery blood by the Griess reaction, mPAP was measured with right ventricular catheter. RESULTS: The level of nitrite in cervical artery blood was significantly increased and the mPAP was markedly reduced after administration of CGRP intravenousely.CONCLUSION: CGRP enhanced the NO level of pulmonary circulation blood and reduces the mPAP significantly in rabbits with ALI.

AIM: To observe the effect of aminoguanidine (AG) on hemodynamics and lung capillary permeability in acute lung injury (ALI) in rabbits. METHODS: 24 rabbits were equally divided into four groups: saline group, endotoxin group, AG group and AG plus endotoxin group. In AG plus endotoxin group, endotoxin was injected to animals to make an ALI model, 25mg/kg AG was injected following that and let this sustain 3 hours. Meanwhile, mean arterial pressure (MAP), mean pulmonary arterial pressures (MPAP) and blood gas analyses were observed during this period. At the end of the experiment, broncho-alveolus lavage was performed, pathologic samples were treated routinely and lung wet weight/dry weight ratio was calculated. RESULTS: After endotoxin injection, MAP and arterial oxygen pressure (PaO 2) decreased, and MPAP increased significantly. The injection of AG had little effect on MAP, but AG could markedly decrease MPAP and increase PaO 2. Cell count in broncho-alveolus lavage fluid (BALF) was less in AG plus endotoxin group than in endotoxin group. Although AG did not affect total protein in BALF, low molecular weight proteins decreased in AG plus endotoxin group by the assay of electrophoresis. Tissue wet weight/dry weight ratio also decreased in this group. Pathologic study showed that there were fewer inflammatory cells and less lung edema in AG plus endotoxin group. CONCLUSION: AG could improve hemodynamics status and attenuate acute lung injury induced by endotoxin in rabbits. [

The phenylacetone monooxygenase, isolated from Thermobifida fusca, mainly catalyzes Baeyer-Villiger oxidation reaction towards aromatic compounds. Met446 plays a vital role in catalytic promiscuity, based on the structure and function of phenylacetone monooxygenase. Mutation in Met446 locus can offer enzyme new catalytic feature to activate C-H bond, oxidizing indole to finally generate indigo and indirubin, but the yield was only 1.89 mg/L. In order to further improve the biosynthesis efficiency of the whole-cell catalyst, metabolic engineering was applied to change glucose metabolism pathway of Escherichia coli. Blocking glucose isomerase gene pgi led to pentose phosphate pathway instead of the glycolytic pathway to become the major metabolic pathways of glucose, which provided more cofactor NADPH needed in enzymatic oxidation of indole. Engineering the host E. coli led to synthesis of indigo and indirubin efficiency further increased to 25 mg/L. Combination of protein and metabolic engineering to design efficient whole-cell catalysts not only improves the synthesis of indigo and indirubin, but also provides a novel strategy for whole-cell catalyst development.
Escherichia coli ; genetics ; metabolism ; Glucose ; metabolism ; Indigo Carmine ; metabolism ; Indoles ; metabolism ; Industrial Microbiology ; methods ; Metabolic Engineering ; Metabolic Networks and Pathways ; Protein Engineering

AIM: To investigate the relationship between inflammatory cell infiltration and proto-oncogenes expression in asthma. METHODS: Guinea pigs were used as asthma models challenged by ovoglobulin. Dot-blot, Northern-blot and immunochemical techniques were used to detect the expression of c-fos, c-myc, c-jun and c-sis. Inflammatory cell infiltration was showed by pathologic study.RESULTS: c-fos and c-myc mRNA could not be detected or expressed at very low level in control group. Those were greatly increased after the animals are challenged by ovoglobulin. Immunochemical study showed that Fos, Myc, Jun and Sis expressed at low level in control group, and those were increased after the challenge. There was little inflammatory cell infiltration in control group. Lymphocyte, neutrophil and eosinophil were detected immediately after the challenge, a great number of inflammation cells could be seen after 12-24 h of the challenge. Majority of neutrophil and eosinophil were under mucosa or in epithelium in airway. CONCLUSION: Oncogenes expression had strong relationship with airway inflammation.

Objective To discuss the nursing care for patients with advanced hepatocellular carcinoma (HCC) who are receiving transcatheter arterial chemoembolization (TACE) combined with sorafenib. Methods A total of 23 consecutive patients with advanced HCC who met the inclusion criteria were enrolled in this study. TACE was carried out in all patients. Three-five days after TACE the patients started to orally take sorafenib. During the treatment course, the patients were kept under close observation for adverse reactions and complications. After leaving the hospital the patients were followed up by the extended care team members, and health education as well as appropriate nursing intervention was carried out. Results All patients with advanced HCC took sorafenib orally after TACE. The major adverse events and complications were gastrointestinal adverse reactions (n = 22) and transient liver dysfunction (n = 23). After leaving the hospital all the patients received regular follow- up examination and extended nursing care. Conclusion Usually, the incidence of adverse reactions is higher in patients with advanced HCC after TACE combined with sorafenib treatment. Careful in-hospital observation and extended out-hospital nursing can reduce the incidence of adverse reactions and thus improve the patient’s quality of life.

BACKGROUND: The presence of ischemic penumbra in hypertensive cerebral hemorrhage is the hot spot and still controversial. The value of 4,9-diaza-2, 3, 10, 10-tetramethydodecan-2, 11-dione dioxime (HL91) tagged with 99Tcm on detecting the hypoxic brain tissue surrounding the hypertensive cerebral hemorrhage nidus, which represents the penumbra is still waited for confirmation.OBJECTIVE: To investigate the value of 99Tcm-HL91 single photon emission computed tomography (SPECT)/CT imaging on detecting hypoxic tissue in the patients with hypertensive cerebral hemorrhage.DESIGN: Control study.SETTING: Department of Neurology, Jiangmen Municipal Central Hospital, Guangdong Province.PARTICIPANTS: This series included 22 patients with hypertensive cerebral hemorrhage examined between March 2004 and March 2005 in Jiangmen Municipal Central Hospital. All cases revealed sudden onset of the disease, presented with the history of hypertension. These patients were diagnosed with hypertension after admission. The hemorrhage occurred in the anterior circulation territory in all cases and the volume of hemorrhage ranged from 10 mL to 63 mL. Minimally invasive stereotaxic aspiration was performed in 3 cases, craniotomy debridement in 1 case, and expectant treatment in the remaining cases. The period of time from the symptom onset to the examination was form 12 hours to 1.5 years, including more than 1 month in 5 cases. Control group consisted of 6 cases were clinically diagnosed with melancholia and anxiety disorders. Cerebral hemorrhage and acute cerebral infarction were ruled out by integrated CT scan in these 6 cases.METHODS: All 22 patients with hypertensive cerebral hemorrhage and 6 normal controls underwent 99Tcm-HL91 SPECT imaging and combined with CT scan.MAIN OUTCOME MEASURES: ① Identification of radioactive concentrations at one side of the peripheral zone of the lesions by visual analysis on two consecutive slices at two different axial directions were considered aspositive hypoxic imaging. ② The other was ROI semi-quantification measuring radiocounting ratio (R) between the region of visible radioactive concentrations, the center of the nidus, and their contralateral mirror region. R ＜ 0.8 or R ＞ 1.2 was considered to be abnormal. ③ Hypoxic region was defined by integrated CT fused imaging, and its volume was calculated using Xelerix workstation. The volume of the hypoxic tissue and hemorrhage was computed by Duotian formula: length of the maximum cross-section of the hemorrhage × width × slice number × 1/2.RESULTS: All 28 patients were involved in the final analysis. ① Perihemorrhagic radioactive concentrations which represented positive hypoxic imaging was revealed on 99Tcm-HL91 SPECT imaging in 18 cases out of 22 patients with cerebral hemorrhage, and positive rate was 77.78%. Bilateral cerebral hemisphere showed symmetric negative imaging in 6 cases of the control group. ② The fused SPECT/CT images revealed hypoxic region was around the intracerebral hemorrhage, small portion was within the nidus of hemorrhage with irregular shape. R value was 1.75±0.10 in perihemorrhagic hypoxic region in 18 cases with positive imaging, and R value was 1.05±0.11 in the basal ganglia in the control group. There was statistically significant difference between the two groups (P ＜ 0.01). ③ There was a positive linear correlation between maximum volume of hematom and hypoxia volume (correlation coefficient: r=0.7517, P ＜ 0.01).CONCLUSION: Relying on the mechanism about demonstrating the hypoxic tissue on fused SPECT/CT imaging, the hypoxic tissue would represent the penumbra may exist in the territories located around the cerebral hemorrhage. The positive territories may be reversible, I.e. The important portion of the penumbra. 99Tcm-HL91 SPECT/CT imaging can detect the hypoxic tissue surrounding the cerebral hemorrhage. The volume of hypoxic tissue is correlated with the hemorrhagic volume. The procedure is promising and could be applied in clinic.

Objective To study the effect of basic fibroblast growth factor(bFGF) on neuron-like differentiation of superparamagnetic iron oxide nanoparticles (SPIONs)-labeled amniotic membrane-derived mesenchymal stem cell.Methods Cells were cultured from enzymatic-digested amniotic membrane tissue.After that,the following steps were taken:(1) Mesenchymal stem cells derived from amniotic membrane were identified by using cell morphology,MTT method and flow cytometry.(2)SPIONs were used to label amniotic membrane-derived mesenchymal stem.(3)bFGF was imported to induce the neuron-like differentiation of SPIONs-labeled amniotic membrane-derived mesenchymal stem cell.Results (1) Primary cultures of P3,amniotic membrane-derived mesenchymal stem cell were fibroblast-like and expression of surface molecules CD29,CD44,CD90 and CD105 was detected,while expression of CD31,CD34,CD45 and CD106 was negative.(2) SPIONs of no more than 14.0 μg/ml are safe to label amniotic membrane-derived mesenchymal stem cells.Cell activity is more than 80％ and expression of surface molecules CD29,CD44,CD90 and CD105 is positive.(3)RT-PCR and immunocytochemistry analysis showed that 10.0 ng/ml bFGF induced neuron-like differentiation of amniotic membrane-derived mesenchymal stem cell (14 μg/ml SPIONs-labeled).Conclusions Enzymatic digestion and cell adherent culture method can be used to isolate mesenchymal stem cells from amniotic membrane.SPIONs of no more than 14.0 μg/ml are safe to label amniotic membrance-derived mesenchymal stem cells and have no effect on the cell activity.Neuron-like differentiation of amniotic membrane-derived mesenchymal stem cell can be induced with 10.0 ng/ml bFGF.

AIM: To observe the antiproliferative effect of c-myc antisense oligonucleotide in rat thymus lymphocytes. METHODS: Rat thymus lymphocytes were separated by Ficoll-Urografin density gradient centrifugation. Lipofectin was used to introduce antisense, sense and mismatched oligonucleotides for c-myc to rat thymus lymphocytes. The antiproliferative effect was assayed by incorporation of -TdR and MTS cell proliferation assay. TR-PCR was used to detect the expression of c-myc mRNA. RESULTS: c-myc antisense oligonucleotide inhibited rat thymus lymphocytes proliferation [(0.14?0.03)A vs (0.32?0.16)A, P

AIM: To observe the inhibitory effect of antisense eukaryotic expression vectors for c-myc on rat airway smooth muscle cells. METHODS: Antisense and sense eukaryotic expression vectors for c-myc pcDNA3-myc-antisense and pcDNA3-myc-sense were constructed. Lipofectin was used to introduce antisense and sense eukaryotic expression vectors for c-myc into rat. The inhibitory effect was assayed by MTT cell proliferation assay. Cell cycles were detected by flow cytometry technology. The expression of c-Myc was detected by immunohistochemistry. RESULTS: The results showed that antisense eukaryotic expression vector for c-myc inhibited rat airway smooth muscle cells proliferation. Rat airway smooth muscle cells were prohibited in S phase and the expression of c-Myc was decreased after antisense eukaryotic expression vectors for c-myc were transfected into cells. CONCLUSION: Antisense eukaryotic expression vectors for c-myc inhibit rat airway smooth muscle cell proliferation. [