Objective:To study the distribution of flurbiprofen axetil ( FA) microemulsion in rats and evaluate the targeting prop-erty to traumatic inflammation tissue. Methods:Traumatic injury model rats were established by hip surgery on the rat right thigh mus-cle and randomly divided into 2 groups:microemulsion group and solution group. Serums and tissues of rats were collected at 15, 30, 60, 120 and 240min after administration (iv. 5 mg·kg-1). A high performance liquid chromatography (HPLC) with UV detection was developed to study the concentration of flurbiprofen in biological samples. Results: The elimination of flurbiprofen axetil microe-mulsion from blood and tissues was slower than flurbiprofen solution. At each 5 time points, drug concentrations of microemulsion group in injury muscle were higher than in normal muscle (P<0. 05), while solution group had no significant difference(P>0. 05). Micro-emulsion group injury muscle Te(targeting efficiency) =12. 21%, and the solution group Te =3. 97%. The Re (relative uptake rate) of injury muscle was 4. 15. Conclusion:The flurbiprofen axetil microemulsion has the targeting property to traumatic injury tissues.

Cell DNA content in heart, liver, kidney of rats were analysed by flow cytometer at different postmortem interved. The rsults show that mean cellular DNA content is 99.5% at 6 hours, 91.3% at 12 hours, 87.1% at 18 hours, 81.3% at 24 hours, 76.7% at 30 hours, 74.3% at 36 hours, 72.3% 48 hours as compared with that at o hours The results ingicates that the quantitative determination of cell DNA in the tissues, mentioned above may provide an objective and reliable approach for the estimation of postmortem time.

Objective To study the phenylpropanoid constituents from the roots of Euphorbia hylonoma Hand-Mazz. Methods The chemical constituents were isolated and purified by silica gel and Sephadex LH-20 column chromatography,and the structures were elucidated on the basis of chemical properties and spectral data. Results Three phenylpropanoid constituents were isolated from the acetone extracts of the roots of Euphorbia hylonoma Hand-Mazz,which were hexadecyl-3-methoxy-4-hydroxybenzeneacrylate Ⅰ,ethyl brevifolincarboxylate Ⅱ and(+) 3′-angeloyl-4′-isovalerylcis-khellactone peuformosin Ⅲ. Conclusion The above compounds were isolated from Euphorbia hylonoma Hand-Mazz for the first time,and the compound Ⅲ was the first obtained from the Euphorbiaceae.

Objective To investigate the efficacy of volume target pressure control(VTPC)and synchronized intermittent mandatory ventilation(SIMV)in treating severe neonatal respiratory distress syndrome(NRDS). Methods Fifty - six admitted cases with severe NRDS hospitalized in Jiangmen Central Hospital from October 2012 to March 2015 were randomly divided into 2 groups:28 cases in VTPC group were treated by VTPC and SIMV,and 28 cases in pressure control ventilation(PCV)group were treated by PCV and SIMV. There was no significant difference between 2 groups in terms of gender,gestational age,and birth weight(all P ﹥ 0. 05). Artery blood gas analysis was performed at 6 hours,12 hours,24 hours,and 48 hours respectively after ventilation. The following parameters were observed:the time of invasive mechanical ventilation,duration of oxygen therapy,mortality and the incidence rates of hypocapnia,pneumo-thorax,ventilator associated pneumonia( VAP),grade Ⅲ - Ⅳ periventricular intraventricular hemorrhage( PVH -IVH),periventricular leukomalacia(PVL)and bronchopulmonary dysplasia(BPD). Results No case in 2 groups withdrew from the test. There was no significant difference between 2 groups in terms of the first treatment time and total doses of poractant alfa injection(all P ﹥ 0. 05). The time of invasive mechanical ventilation in VTPC group[(71. 75 ± 9. 82)h]was shorter than that in PVC group[(97. 89 ± 16. 88)h](t = 7. 083,P = 0. 000). Hypocapnia incidence of four blood gas analysis in VTPC group[(19. 64 ± 14. 20)% ]was lower than that in PCV group[(47. 32 ± 18. 43)% ] (t = 6. 294,P = 0. 000). Incidence rates of VAP and PVL in VTPC group were lower than those in PCV group(χ2 =5. 197,P = 0. 023;χ2 = 4. 766,P = 0. 029). However,duration of oxygen therapy,mortality and the incidence rates of pneumothorax,Ⅲ - Ⅳ PVH - IVH and BPD were not significantly different between 2 groups( all P ﹥ 0. 05). Conclusion VTPC + SIMV has a better efficacy than PCV + SIMV in the treatment of NRDS.

ObjectiveTo observe effects of Naofucong grain on learning and memory in cerebral ischemic mice, and identify the mechanisms on N-methyl-D-aspartate (NMDA) receptor.MethodsExperimental dementia mice were induced by ischemia reperfusion injury, and treated with Naofucong grain or Dangguishaoyao powder (positive control medicine). Learning and memory were tested in the Morris water maze. Activities of NMDA receptor in brain cortex and hippocampus were detected ResultsIncubation periods of mice treated with Naofucong grain were significantly shorter than that of ischemic model mice (P<0.01) Activities of NMDA receptor in brain cortex and hippocampus showed no difference between Naofucong grain treated mice and normal mice (P>0.05)Activities of NMDA receptor in brain cortex and hippocampus in Naofucong grain treated mice were significantly lower than that in ischemic model mice(P<0.01) ConclusionThe improvement of Naofucong grain in the memory and learning in cerebral ischemic mice may be related to the inhibition of the activity of NMDA receptor.

ObjectiveTo examine effects of traditional Chinese herbs Naofucong grain on PC12 cells injured by H2O2 in vitro MethodsRats random divided into 2 groups: Naofucong grain treating group (administered with Naofucong grain) and control group (administered with distilled water) After 3 days administration, serums were obtained and added to cultured PC12 cells which were under different concentrations of H2O2 Morphological examinations in PC12 cells were detected by MTT measurement ResultsMTT measurement showed that Naofucong grain administered rats serums significantly increased the survival number of PC12 cells in different concentrations of H2O2.Conclusion Naofucong grain may have obvious protective effects on PC12 cells injured by H2O2.

Hypermethylation in the promoter region is an important epigenetic mechanism for the transcriptional repression of a number of cancer-associated genes, and over-expression and/or increased activity of DNA methyltransferases are considered to be the main cause of promoter hypermethylation. In order to explore the roles of two methyltransferase members (DNMT1 and DNMT3b) in the cholangiocarcinoma tumorigenesis, antisense eukaryotic expression plasmid of DNMT1 and DNMT3b gene was constructed respectively, and were co-transfected into the human cholangiocarcinoma cell line QBC-939 to observe their biological effects on the cell growth and proliferation ability, apoptosis, cell cycle alteration, and the tumorigenesis ability in the subcutaneous tissue of nude mouse. The results demonstrated that co-transfection with antisense eukaryotic expression plasmid of DNMT1 and DNMT3b gene and single transfection with antisense eukaryotic expression plasmid of DNMT1 gene can suppress the growth and proliferation of QBC-939, block the cell cycle at G1 phase, increase the apoptosis rate, minimize the tumor size in the subcutaneous tissue of nude mouse. The suppressing biological effect of co-transfection is stronger than single transfection with antisense DNMT1. Meanwhile, single transfection with antisense eukaryotic expression plasmid of DNMT3b gene has no effects on the biological characteristics of QBC-939. This study suggests that DNMT1 gene plays a key role in DNA methylation and DNMT3b gene may act as an accessory to support its function in inactivation of tumor suppressor genes. Combination DNMT1 and DNMT3b will increase their biological effects and have the synergistic effect on suppressing the growth of human cholangiocarcinoma cell line QBC-939.
Apoptosis ; Biliary Tract Neoplasms/*metabolism ; Cell Line, Tumor ; Cell Proliferation ; Cholangiocarcinoma/*metabolism ; DNA (Cytosine-5-)-Methyltransferase/*biosynthesis ; DNA (Cytosine-5-)-Methyltransferase/genetics ; Down-Regulation ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; Mice, Nude ; Neoplasm Transplantation

Objective To investigate the apoptosis of articular chondrocyte and the expression of Bcl-2, Bax, Fas and iNos in articular cartilage with Kashin-Beck disease(KBD) in order to understand the pathogenesis of chondronecrosis in KBD. Methods The collected samples of human articular cartilage were divided into two groups: control group (15 samples from 15 cases), KBD group (15 samples from 15 cases). KBD patients were diagnosed by "Pathological Criteria to Diagnose KBD in China". Chondrocyte apoptosis was detected by TUNEL staining, and the Bcl-2, Bax, Fas and iNos positive articular chondrocytes were stained by the B-SA of immunohistochemistry. Articular cartilage was classified three zones and the positive rate were counted by light microscope for cytoplasimic staining by polyclonal antibodies of Bcl-2, Bax, Fas and iNos and apoptotic chondrocytes by TUNEL. Results ① The percentage of positive apoptotic chondrocytes stained by TUNEL in the middle zone of articular cartilage from the KBD-children group(33.60±2.71%) was higher than that of the control (1.33±0.41% t=11.59, g=28, P<0.01). ②The percentage of chondrocytes staining for Bcl-2, Bax, Fas and iNos among the upper and the middle zone in KBD group were significantly higher than that of the control (t=11.75-18.65, g=14, P<0.01); the remarkable difference in the expression of Bcl-2, Bax, Fas and iNos among the upper, the middle and the deep zones was also seen in KBD articular cartilage (F=73.49-114.42, g=42, P<0.01), and staining for Bcl-2, Bax, Fas and iNos in KBD children was prominent in the upper zone(41.93±12.26%, 45.60±15.78%, 53.60±16.49%, 45.47±14.02%) and the middle zone(14.93±3.50%, 13.87±4.32%, 23.27±4.83%, 21.67±6.82%)of articular cartilage, respectively. Conclusion The chondrocyte apoptosis and the present of Bcl-2, Bax, Fas and iNos positive chondrocytes in articular cartilage of children with KBD were significantly higher than that of the control.

Objective To study the effect of transfection of antisense MBD1 gene eukaryotic expression vector on the expression of MBD1 gene in human cholangiocarcinoma cell line QBC-939.Methods The(constructed) antisense MBD1 gene eukaryotic expression vector was transfected into the human(cholangiocarcinoma) cell line QBC-939 using lipofectamine transfection reagents,and positive cell clones were obtained using G418 selection after transfection.The constructed recombinant vector was transfected into(QBC-939) cells successfully and was confirmed by amplifying the exogenous neo~R gene with PCR method.The expression level of MBD1 gene mRNA and protein was detected by RT-PCR and FCM methods respectively.Results Following the transfection,the MBD1 gene mRNA level in human cholangiocarcinoma cell line QBC-939 decreased from 0.912?0.022 to 0.215?0.017,and the MBD1 gene protein level also(decreased) from(80.19?5.05)% to(35.11?4.05)%.There were very significant differences on the expression both at the transcription and post-transcription levels of MBD1 gene between non-tranfection group and the antisense MBD1 gene eukaryotic expression vector transfection group(P

Objective To investigate the effects of the tumor suppressor gene PTEN in growing inhibition and down-regulating mTOR in cholangiocarcinoma QBC939 cells in vitro.Methods QBC939 cells were transfected with plasmids wild-type PTEN and C124S-PTEN in vitro.After transfection,the expression of the PTEN and phosphorylation of AKT and mTOR was detected by Western blot.Flow cytometry was used to analyze apoptosis and cell cycle of the transfected cells.Results Compared with the control,the expression of phosphorylation AKT was decreased and mTOR were down-regulated respectively when transfected with the wild-type PTEN.However,after transfection with mutation-type PTEN,the level of PTEN in the cells by increased,but phosphorylation AKT level and mTOR expression had no significant change.Conclusions PTEN can be actived by phosphorylated AKT.Actived AKT decreased the mTOR which led to tumor cells apoptosis and regulation of the tumor cell cycle.In the pathway of signal transmission of PI3K/AKT/PTEN/mTOR,PTEN and mTOR are closely related through phosphorylation of AKT.