Applications of zebrafish xenotransplantation models for in vivo evaluation of anticancer drug sensitivity test / 中国比较医学杂志

Objective To evaluate the sensitivity to 5 clinically commonly used anticancer drugs in vivo using the zebrafish xenotransplantation models of human lung cancer,stomach cancer,and liver cancer cells,respectively. Methods Zebrafish xenotransplantation models of A549 lung cancer cells,SGC-7901 stomach cancer cells and HepG2 liver cancer cells were established. The xenograft models of A549 cells were treated with three different doses of cis-platinum, paclitaxel, vinorelbine, endostar and bevacizumab, respectively. The SGC-7901 model was treated with three concentrations or doses of paclitaxel, irinotecan, hydroxyurea, cis-platinum and 5-fluorouracil, respectively. And the HepG2 model was treated with three concentrations or doses of adriamycin,gemcitabine,hydroxyurea,cis-platinum and 5-fluorouracil. The tumors were analyzed and quantified in vivo by fluorescence microscopy,and the inhibition rates of tumor growth with each drug were calculated and compared with the model control group for statistical significance. Results All of the tested anticancer drugs showed inhibitory effect on tumor cells in the zebrafish xenograft models with statistical significance in a dose-dependent manner. During the drug sensitivity test,the inhibition rate of bevacizumab on A549 lung cancer cells decreased in the order(65%)> cis-platinum(55%)> vinorelbine(40%)> endostar(39%)>paclitaxel(27%). As for the SGC-7901 stomach cancer cells, the tumor growth inhibition rate decreased in the order hydroxyurea(46%)> 5-FU(31%)= irinotecan(31%)> paclitaxel(26%)> cis-platinum(24%). And the therapeutic effect of cis-platinum on the HepG2 liver cancer cells decreased in the order(64%)> hydroxyurea(56%)>gemcitabine(46%)> adriamycin(45%)> 5-FU(38%). Conclusions Zebrafish xenotransplantation models of cancer cells are suitable for in vivo sensitivity test of anticancer drugs.

Effects of calcitonin gene-related peptide on aquaporin 1 and aquaporin 5 expressions in young rats with acute lung injury / 中华实用儿科临床杂志

Objective To explore the effect of calcitonin gene-related peptide (CGRP) on expressions of aquaporin (AQP)1 and AQP 5 in young rats with acute lung injury (ALI) caused by lipopolysaccharide (LPS).Methods Eighty-four young rats were randomly divided into control group,ALI model group and CGRP group.The rats in ALI model group were given intraperitoneal injection of LPS (5 mg/kg)for 2,6,12,24 hours;while the rats in CGRP group were given intraperitoneal CGRP (1 mg/kg) after 1 h injection of LPS.At 2 h,6 h,12 h and 24 h,all rats were sacrificed and lung tissues were obtained.The histopathological changes in lung tissues were evaluated by adopting hematoxylin-eosin staining,and wet/dry(W/D) was measured.The mRNA and protein levels of AQP1 and AQP5 in lung tissues were detected by adopting fluorogenic quantitative PCR (qPCR) and Western blot.Results Pathological stain showed that rats in control group had a normal lung tissue structure,and LPS made lung tissue edema,narrowing the alveolar cavity and inflammatory cell infiltration.CGRP attenuated the effect of LPS on rat's lung.The W/D ratio of lung tissue was significantly higher than that in the control group,and CGRP reduced the W/D ratio of lung tissue.qPCR showed that the mRNA levels of AQP1 and AQP5 from rats in ALI group (0.009 ±0.001 and 0.055 ±0.006)decreased compared with those in the control group (0.035±0.002 and 0.167 ±0.006) and CGRP group (0.024 ± 0.002 and 0.134 ± 0.012) (all P ＜ 0.001).Western blot results showed after 24 h injection of LPS,both AQP1 and AQP5 levels from ALI group (0.397 ± 0.041 and 0.215 ± 0.029) were significantly lower than those in the control group (0.850 ± 0.020 and 0.741 ± 0.032) (all P ＜ 0.001),and their levels in CGRP group (0.593-± 0.065 and 0.461 ± 0.039) were also lower than those in the control group,but higher than those in ALI group (all P ＜ 0.001).Conclusion CGRP can enhance AQP1 and AQP5 levels and reduce pulmonary edema,and it has a protective effect on rats with acute lung injury.

Effects of calcitonin gene-related peptide on aquaporin 1 and aquaporin 5 expressions in young rats with acute lung injury / 中华实用儿科临床杂志

Objective To explore the effect of calcitonin gene-related peptide (CGRP) on expressions of aquaporin (AQP)1 and AQP 5 in young rats with acute lung injury (ALI) caused by lipopolysaccharide (LPS).Methods Eighty-four young rats were randomly divided into control group,ALI model group and CGRP group.The rats in ALI model group were given intraperitoneal injection of LPS (5 mg/kg)for 2,6,12,24 hours;while the rats in CGRP group were given intraperitoneal CGRP (1 mg/kg) after 1 h injection of LPS.At 2 h,6 h,12 h and 24 h,all rats were sacrificed and lung tissues were obtained.The histopathological changes in lung tissues were evaluated by adopting hematoxylin-eosin staining,and wet/dry(W/D) was measured.The mRNA and protein levels of AQP1 and AQP5 in lung tissues were detected by adopting fluorogenic quantitative PCR (qPCR) and Western blot.Results Pathological stain showed that rats in control group had a normal lung tissue structure,and LPS made lung tissue edema,narrowing the alveolar cavity and inflammatory cell infiltration.CGRP attenuated the effect of LPS on rat's lung.The W/D ratio of lung tissue was significantly higher than that in the control group,and CGRP reduced the W/D ratio of lung tissue.qPCR showed that the mRNA levels of AQP1 and AQP5 from rats in ALI group (0.009 ±0.001 and 0.055 ±0.006)decreased compared with those in the control group (0.035±0.002 and 0.167 ±0.006) and CGRP group (0.024 ± 0.002 and 0.134 ± 0.012) (all P ＜ 0.001).Western blot results showed after 24 h injection of LPS,both AQP1 and AQP5 levels from ALI group (0.397 ± 0.041 and 0.215 ± 0.029) were significantly lower than those in the control group (0.850 ± 0.020 and 0.741 ± 0.032) (all P ＜ 0.001),and their levels in CGRP group (0.593-± 0.065 and 0.461 ± 0.039) were also lower than those in the control group,but higher than those in ALI group (all P ＜ 0.001).Conclusion CGRP can enhance AQP1 and AQP5 levels and reduce pulmonary edema,and it has a protective effect on rats with acute lung injury.

Molecular Mechanism of Action of hnRNP K and RTN3 in the Replication of Enterovirus 71 / 病毒学报

Enterovirus 71 (EV71) is a neurotropic pathogen that can induce hand, foot and mouth disease in children. There is an appreciable mortality rate after EV71 infections. The mechanism of action of EV71 replication is not known. Recent work has identified some of cell factors of the host that participate in the synthesis of the RNA and proteins of EV71 (e.g., hnRNP K, reticulon 3 (RTN 3)). In that work, researchers used a competitive assay to show that hnRNP K can interact with EV71 5' UTR, which is required for efficient synthesis of viral RNA. Using a yeast two-hybrid system, other researchers demonstrated that RTN 3 interacts with the N-terminal domain of EV71 2C, which is crucial for replication of viral RNA. Here, we discuss recent work focusing on the molecular mechanisms of hnRNP K and RTN 3 in the synthesis of the RNA and proteins of EV71.

ClassⅠ integron and its correlation with genes coding forESBLs in ESBLs-producing Escherichia coli from chil-dren / 临床儿科杂志

ObjectiveTo study the correlation with Genes Coding forESBLs and ClassⅠIntegron in ESBLs-producing Escherichia coli from children.MethodsPCR was used for gene typing detection of 100 strains of ESBLs-producingEsche-richia colistrains. While detection of class I integrase gene in the 100 strains ESBLs-producingEscherichia coli and 100 strains of non-ESBLs producingEscherichia coli were separately performed by PCR. It provides the solid base not only to reveal the relationship between class I integron and drug resistance, but also the relationship between class I integron and ESBLs-producing. ResultsThe most frequently genotyping from ESBLs-producingEscherichia coli in children isCTX-M (84%), followed by TEM-1(63%). The predominant distribution of genotype in ESBL- producing strains isTEM-1 +CTX-M (45%), followed by CTX-M (34%). Class I integrase gene detected in ESBLs- producing and non- ESBLs producing strain were 100 cases (100%) and 25 cases (25%) separately, the difference was statistically signiifcant (P<0.05); drug resistance in class I integron positive strains were signiifcantly higher than in class I integron negative strains, especially in Ciprolfoxacin, Levolfoxacin, and Amino-glycoside (86.4%, 88%, and 80%).ConclusionsThe distribution of Class I integron in ESBLs-producingEscherichia coli is signiifcantly higher than that in non-ESBLs-producing strains, It is rational that Class I integron highly correlate with strong drug resistance in ESBLs-producing strains.

Evaluation on the efficacy and safety of domestic bivalirudin during percutaneous coronary intervention / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Bivalirudin was widely used as an anticoagulant during coronary interventional procedure in western countries. However, it was not available in China before this clinical trial was designed. This randomized, single-blind and multicenter clinical trial aimed to evaluate the efficacy and the safety of domestic bivalirudin during percutaneous coronary intervention (PCI).</p><p><b>METHODS</b>A randomized, single-blind, multicenter trial was designed. Elective PCI candidates in five centers were randomized into a bivalirudin group and a heparin group, which were treated with domestic bivalirudin and non-fractional heparin during the PCI procedure. The efficacy was evaluated by comparing the activated coagulation time (ACT), the procedural success rate (residual stenosis < 20% in target lesions without any coronary artery related adverse events within 24 hours after PCI), and the survival rate without major adverse cardiac events at 30 days after PCI between the two groups. Safety was evaluated by the major/minor bleeding rate.</p><p><b>RESULTS</b>A total of 218 elective PCI patients were randomized into a bivalirudin group (n = 110) and heparin group (n = 108). Except for two patients needing additional dosing in the heparin group, the ACT values of all other patients in both groups were longer than 225 seconds at 5 minutes after the first intravenous bolus. Procedural success rates were respectively 100.0% and 98.2% in the bivalirudin group and heparin group (P > 0.05). Survival rates without major adverse cardiac events at 30 days after PCI were 100.0% in the bivalirudin group and 98.2% in the heparin group (P > 0.05). Mild bleeding rates were 0.9% and 6.9% (P < 0.05) at 24 hours, and 1.9% and 8.8% (P < 0.05) at 30 days after PCI in the bivalirudin group and heparin group respectively. There was one severe gastrointestinal bleeding case in the heparin group.</p><p><b>CONCLUSIONS</b>Domestic bivalirudin is an effective and safe anticoagulant during elective PCI procedures. The efficacy is not inferior to heparin, but the safety is superior to heparin.</p>

Biochemical screening and genetic diagnosis of thalassemia in children from Kunming / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the types and frequency of gene mutations in children with thalassemia in Kunming, Yunan Province.</p><p><b>METHODS</b>A biochemical screening for thalassemia was performed by testing RBC fragility, MCV and hemoglobin electrophoresis on 1338 children from Kunming, Yunnan Province. Genetic diagnosis was performed on the children with α-thalassemia by gap-PCR and on the children with β-thalassemia by PCR-RDB.</p><p><b>RESULTS</b>The positive rate of the biochemical screening for thalassemia was 11.36% (152 cases). The positive rate of genetic diagnosis was 8.59% (115 cases). Of the 115 cases, α-thalassemia was found in 43 cases, β-thalassemia in 68 cases and α-combined-β thalassemia in 4 cases.--SEA/αα accounted for 47%, -α4.2/αα accounted for 21%, and HbH disease accounted for 14%. Six genotypes were found in 68 cases of β-thalassemia and the mutation frequency of βE was the highest (32%), followed by CD41-42 (24%), CD17 (23%), IVS-II654 (10%), CD71-72 (10%), and -28 (1%).</p><p><b>CONCLUSIONS</b>The frequency of gene mutations for thalassemia is high in children from Kunming, Yunnan Province. Premarital and prenatal screenings and genetic diagnosis for thalassemia should be carried out in this area.</p>

Epidemiological analyses of children with mycoplasma pneumoniae infection in kunming region between 2003 and 2007 / 中国小儿急救医学

Objective To investigate the epidemiological characteristics of children infected by mycoplasma pneumoniae(MP)over the last five years in Kunming region.Methods Indirect immunofluorescence assay was used to detect blood MP-IgM of hospitalized patients from January 2003 to December 2007 in order to determine the age and gender distribution characteristics of MP infection and to investigate the epidemiological features of the five years' results.Results Infection incidences by MP in Kunming region over the last five years were 20.9%,14.3%,17.5%,15.7%and 19.5%,respectively.Statistical significance was found among the groups mentioned above(P＜0.01).The MP infection incidences in different age groups were 10.7%(～1 year old),20.5%(～3 years' old),21.5%(～6 years' old)and 21.7%(～14 years' old).MP infection incidences showed age and gender characteristics(P＜0.01).Infants showed lower NIP infection incidence and infection incidence was higher in female.The epidemiological investigation indicated that MP infection incidences varied from year to year(P＜0.01).The infection incidences of 2003 and 2007 by MP were higher than those of the other years.There were epidemiological differences in infection incidences by NIP over last five years(summer,autumn and winter).The MP infection incidences showed seasonal differences(P＜0.01).Conclusion Mycoplasma pneumoniae was the main local causative agent responsible for respiratory tract infection in children aged under one years' old in Kunming region.Over the last five years,there were two outbreaks in the local area,but with no seasonal regularity of epidemiology.

EphrinB2 gene transfection promotes the differentiation of bone marrow mesenchymal stem cells into vascular endothelial cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the effects of ephrinB2 gene transfection on the differentiation of bone marrow mesenchymal stem cells (BMSCs) into vascular endothelial cells.</p><p><b>METHODS</b>Wistar rat BMSCs were isolated by density gradient centrifugation and purified on the basis of their adhesion ability. The BMSCs were transfected with a lenti-virus vector encoding a constitutively active form of human ephrinB2 gene, and the cell markers including CD105, CD73, CD44, von Willebrand factor (VWF) and vascular growth factor receptor 2 (KDR) were detected using flow cytometry. The potential of ephrinB2-BMSCs for differentiation into osteoblasts and adipoblasts in vitro were tested, and the differentiation of the cells into endothelial-like cells was induced by culture in the presence of 2% fetal bovine serum and 50 ng/ml vascular endothelial growth factor.</p><p><b>RESULTS</b>EphrinB2-BMSCs were positive for the markers CD105, CD73 and CD44, and negative for the typical endothelial markers like VWF and KDR, and retained high potentials for differentiation into osteoblasts and adipoblasts in vitro after cultivation in respective media. After induced differentiation, ephrinB2-BMSCs expressed VWF and KDR and showed greater ability of differentiation into vascular endothelial cells and formation of capillary structures on matrix gel than the BMSCs without transfection.</p><p><b>CONCLUSIONS</b>EphrinB2 gene transfection efficiently promotes the differentiation of BMSCs into vascular endothelial cells. These genetically engineered cells provide valuable sources for new therapies of coronary heart disease.</p>

Early Diagnostic Value of Reverse Transcription Polymerase Chain Reaction Combined with Indirect Immunofluorescence Method for Atypical Pathogens Infection of Lower Respiratory Tract in Children / 实用儿科临床杂志

0.05).While the diagnosis ratio of MP and LP with RT-PCR+IIF method were higher than any of other 3 methods significantly(Pa