Chemical Constituents of n-Butanol Extract Part of Akebia trifoliata Caulis / 中国实验方剂学杂志

<b>Objective::To study the chemical constituents from <italic>n</italic>-butanol extract of <italic>Akebia trifoliata</italic> caulis. <b>Method::The 100 kg caulis of <italic>A</italic>. <italic>trifoliata</italic> was extracted with 75% ethanol (EtOH) for three times by heating reflux. These 3 extracts were decompressed and concentrated, and then dissolved in water. The solvent was successively extracted with dichloromethane, ethyl acetate and <italic>n</italic>-butanol. The chemical constituents from the <italic>n</italic>-butanol fraction were isolated by macroporous, silica gel, sephadex LH-20 and ODS columns, and semi-preparative high performance liquid chromatography, and their chemical structures were determined through MS, NMR analysis (¹H and ¹³C-NMR) and spectroscopic data from literatures. <b>Result::Totally 14 compounds were isolated and identified as mutongsaponin B(<bold>1</bold>), mutongsaponin C(<bold>2</bold>), saponin PH(<bold>3</bold>), begoniifolide A(<bold>4</bold>), 2<italic>α</italic>, 3<italic>β</italic>, 23-trihydroxy-30-noroleana-12, 19-dien-28-oicacid-<italic>O</italic>-<italic>β</italic>-<italic>D</italic>-xylopyranosyl-(1→3)-<italic>α</italic>-<italic>L</italic>-rhamnopyranosyl-(1→4)-<italic>β</italic>-<italic>D</italic>-glucopyranosyl-(1→6)-<italic>β</italic>-<italic>D</italic>-glucopyranosyl ester(<bold>5</bold>), akemisaponins D(<bold>6</bold>), akemisaponins E(<bold>7</bold>), asiaticoside(<bold>8</bold>), saponin PJ1(<bold>9</bold>), scheffoleoside A(<bold>10</bold>), symplocosneolignan A(<bold>11</bold>), kalopanax-saponins D(<bold>12</bold>), leonticin E(<bold>13</bold>), ciwujianoside A₁(<bold>14</bold>). <b>Conclusion::Compounds <bold>1-4</bold>, <bold>11</bold>, <bold>13, 14</bold> were isolated from this plant for the first time. The discovery of these compounds further enriched the chemical constituents of <italic>A</italic>. <italic>trifoliata</italic>, and provided experimental and scientific basis for the comprehensive development and utilization of <italic>A</italic>. <italic>trifoliata</italic>.

A retrospective analysis of 6 children with Duchenne muscular dystrophy / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To analyze the clinical features of 6 children with Duchenne muscular dystrophy (DMD) and review related literature, and to provide a basis for early diagnosis and effective treatment of this disease.</p><p><b>METHODS</b>A retrospective analysis was performed on the clinical data of 6 children with DMD who were admitted to the First Affiliated Hospital of Nanjing Medical University from January 2010 to October 2015.</p><p><b>RESULTS</b>All the 6 cases were boys without a family history of DMD, and the age of diagnosis of DMD was 1.2-11.5 years. All patients had insidious onset and increases in alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase, α-hydroxybutyrate dehydrogenase, creatine kinase (CK), and creatine kinase-MB, particularly CK, which was 3.3-107.2 times the normal level. Their gene detection results all showed DMD gene mutation. The gene detection results of two children's mothers showed that they carried the same mutant gene. The muscle biopsy in one case showed that the pathological changes confirmed the diagnosis of DMD. The level of CK in one case declined by 77.0% 5 days after umbilical cord blood mesenchymal stem cell transplantation.</p><p><b>CONCLUSIONS</b>For boys with abnormal serum enzyme levels and motor function, DMD should be highly suspected. It should be confirmed by CK and DMD gene detection as soon as possible. And the progression of the disease could be delayed by early intervention for protecting the remaining normal muscle fibers.</p>

Roles of histone acetyltransferase and histone deacetylase in the pathogenesis of bronchial asthma / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To evaluate the roles of various cytokines, histone acetyltransferase (HAT) and histone deacetylase (HDAC) in the pathogenesis of bronchial asthma.</p><p><b>METHODS</b>BALB/C mice were randomly assigned to control, untreated asthma, hormone treatment and TSA treatment groups. Bronchial asthma was induced by intraperitoneal injections and atomization inhalation of ovalbumin (OVA) in the asthma, hormone treatment and trichostatin (TSA) treatment groups. The mice in the hormone treatment and TSA treatment groups were administered with dexamethasone 1.0 mg/kg and TSA 1.0 mg/kg respectively by an intraperitoneal injection 30 minutes before challenge of asthma. At 24 hours after the last challenge, IL-4, IL-8 and IFN- levels in serum were measured using ELISA, and activities of HAT and HDAC in peripheral blood mononuclear cells (PBMC) were determined by the enzyme linked immunofluorescence assay.</p><p><b>RESULTS</b>The serum levels of IL-4 and IL-8 in the untreated asthma group were higher than in the control, hormone treatment and TSA treatment groups (P<0.05). There was no difference in the serum levels of IL-4 and IL-8 among the control, hormone treatment and TSA treatment groups (P>0.05). The activity of HDAC in the untreated asthma group was lower than in the control, hormone treatment and TSA treatment groups (P<0.05). Hormone treatment significantly decreased the activity of HAT compared with the untreated asthma group (P<0.05). There was no difference in the activities of HAT and HDAC among the control, hormone treatment and TSA treatment groups (P>0.05).</p><p><b>CONCLUSIONS</b>The decreased activity of HDAC leads to an increased secretion of inflammatory factors and thus induces asthma.</p>

Effects of small interfering RNA (siRNA) against Par-4 gene on the apoptosis of human bone marrow mesenchymal stem cells / 中华儿科杂志

<p><b>OBJECTIVE</b>The prostate apoptosis response factor-4 (Par-4) gene was originally identified by differential screening for genes that are up-regulated when prostate cells are induced to undergo apoptosis. Par-4 was found to possess potent apoptotic activity in various cellular systems in response to numerous stimuli. The aim of this study was to explore the effects of small interfering RNA (siRNA) against Par-4 gene on the apoptosis of human bone marrow mesenchymal stem cells (hBMSCs) exposed to glutamate.</p><p><b>METHODS</b>Primary culture of hBMSCs was carried out and siRNAs targeted Par-4 gene (Par-4-SiRNA) were chemically synthesized. Eukaryocytic expression vector was built and were transfected into hBMSCs with liposome. After selecting with G418, the stable cell clones were treated with glutamate. The expression of Par-4 mRNA was determined by real-time PCR. The apoptosis of hBMSCs was quantified by flow cytometry. Western blotting was used to detect the protein levels of phosphorylated Akt1 (Thr308). Relative Caspase-3 activity was determined by colorimetric assay.</p><p><b>RESULTS</b>The Par-4-SiRNA-1 and Par-4-siRNA-2 could markedly down-regulate the mRNA levels of Par-4 gene in hBMSCs. With the transfections of Par-4-SiRNA-1 and Par-4-SiRNA-2, the levels of Par-4 mRNA were respectively decreased by 88% and 67%. Both Par-4-SiRNA-1 and Par-4-SiRNA-2 inhibited significantly the apoptosis of hBMSCs induced by glutamate, in which the percentages of apoptotic cells were respectively decreased to 38.80% +/- 3.97% (P < 0.01) and 45.49% +/- 4.32% (P < 0.01) from 60.30% +/- 6.82%. Western blot assays demonstrated that, glutamate down-regulated the expression of phosphorylated Akt1 proteins in hBMSCs (89.07 +/- 6.42 and 28.30 +/- 5.65, respectively, P < 0.01). However, Par-4-SiRNA-1 and Par-4-SiRNA-2 could markedly recover the down-regulation of Akt1 proteins induced by glutamate (63.56 +/- 6.75 and 45.59 +/- 4.88, respectively, P < 0.01). And the relative Caspase-3 activity which was enhanced by the treatment with glutamate (0.1428 +/- 0.0495 and 0.8616 +/- 0.1051, P < 0.01), was suppressed by Par-4-SiRNA-1 and Par-4-SiRNA-2 (0.8616 +/- 0.1051 and 0.6581 +/- 0.0555, respectively, P < 0.01).</p><p><b>CONCLUSION</b>SiRNA against Par-4 gene could inhibit the apoptosis of hBMSCs induced by glutamate, and its inhibitory effects may be mediated by the up-regulation of phosphorylated Akt1 and the suppression of the relative Caspase-3 activity.</p>

Comparative study on different point combinations of Zusanli (ST 36) for improving acute gastric mucosa injury / 中国针灸

<p><b>OBJECTIVE</b>To compare effects of different point combinations of Zusanli (ST 36) for improving acute gastric mucosa injury and study on the mechanism.</p><p><b>METHODS</b>One hundred rats were randomly divided into 10 groups, Zusanli (ST 36) group (group A); Zusanli and Neiguan (PC 6) group (group B); Zusanli and Zhongwan (CV 12) group (group C); Zusanli and Gongsun (SP 4) group (group D); Zusanli, Neiguan and Zhongwan group (group E); Zusanli, Neiguan and Gongsun group (group F); Zusanli, Zhongwan and Gongsun group (group G); Zusanli and Neiguan, Zhongwan, Gongsun group (group H); model group (group I); blank control group (group J), 10 rats in each group. Gastric mucosa injury model was made by intragastric infusion of dehydrated alcohol (0. 6 mL/100 g). The gastric mucosa injury index (UI), epidermal growth factor (EGF), nitric oxide (NO) and gastrin (GAS) contents were detected.</p><p><b>RESULTS</b>Contents of EGF and NO were significantly increased and GAS content decreased in all of the EA groups as compared with those in the model group (P<0. 01 or P<0. 05), with no significant differences among group A, B and D, and significant differences as group A compared with group C, F and group H compared with other EA groups.</p><p><b>CONCLUSION</b>Different point combinations of Zusanli (ST 36) can improve acute gastric mucosa injury, with the strongest effect in the Zusanli and Neiguan, Zhongwan, Gongsun group.</p>

Expression of Cardiotrophin-1 in Myocardium and Peripheral Blood Plasma of Neonatal Rat with Asphyxia and Regulative Effect of Neuregulin-1 / 实用儿科临床杂志

Objective To explore the expression of cardiotrophin-1(CT-1) in myocardium and peripheral blood plasma of neonatal rat with asphyxia and the regulative effect of neuregulin-1(NRG-1).Methods Ninety seven-day-old neonatal rats were randomly divided into 3 groups:asphyxia group (n=40),normal control group (n=10)and NRG-1 pretreatment group (n=40).The model of neonatal rat with asphyxia was prepared by the way of ligation of carotid combined with low supply of oxygen.NRG-1(1 mg/kg) was given to NRG-1 pretreatment group by intraperitoneal injection 30 min before asphyxia.The separated plasma of peripheral blood and myocardium antetheca of aortic ventricle of heart were taken at the time point of 6,12,24 and 48 h.The expression of CT-1 in peripheral blood plasma was detected by enzyme linked immunoadsorbent assay,and that of myocardium was determined by Western blot.Results The expressions of CT-1 protein in peripheral blood plasma of asphyxia group were significantly higher than those of normal control group at each time point,and reached the peak at 24 h(Pa

Isolation and Identification of Promoter Sequence of Interferon Regulatory Factor-3 and Detection of Its Promotor Activity in Human Embryonic Kidney-293 Cells / 实用儿科临床杂志

Objective To construct a luciferase reporter plasmid containing interferon regulatory factor 3(IRF-3)human gene promoter and to evaluate promoter activity in human embryonic kidney(HEK)-293 cells.Methods The 1 000 bp fragment was amplified by PCR with human genomic DNA as a template and was directionally cloned into pGL3-basic multiple cloning sites to construct the luciferase repor-ter plasmid pGL3-pIRF-3.Transfection of HEK-293 cells with the promoter-driven lucife-rase construct was performed to induce lucife-rase gene expression and calculate the relative luciferase activity unit(RLU).Promoter sequence of 1 000 bp upstream of transcription initiation site of IRF-3 was analyzed by using Promoter 2.0 Prediction software.Results DNA sequencing and restriction endonuclease analysis verified the successful construction of the plasmid pGL3-pIRF-3.This IRF-3 promoter exhibited a strong promoter activity with an increase of 42.2-fold of RLU in HEK-293 cells when compared with pGL-3 basic vector.The transfection experiment confirmed that the levels of its activation were significantly higher than that in controls in HEK-293 cells.Function analysis of IRF-3 promoter disclosed seve-ral GATA-1 and specific protein 1(Sp1) sites and E2F in minimal promoter region.Conclusion The plasmid pGL3-pIRF-3 promoter is successfully constructed and has a strong basal promoter activity in HEK-293 cells.

Effect of Licofelone on Expression of Fractalkine Induced by Interleukin-18 in Mesangial Cells / 实用儿科临床杂志

Objective To study the effect of Licofelone,a novel non-steroid anti-inflammatory drug,on the expression of Fractalkine induced by interleukin-18(IL-18) in mesangial cells.Methods Rat mesangial cells were cultured and divided into IL-18 stimulated group,Licofelone-treated group and normal control group.The cells in IL-18 stimulated group were stimulated by 10 ?g/L IL-18 for 24 h.In Licofelone-treated group,ahead of exposure of IL-18 for 24 h,cells were treated with Licofelone in the doses of 10,50 and 100 ?mol/L for 30 min.Additionally,the mesangial cells without treatment of IL-18 and Licofelone were used as normal control group.Reverse transcription-polymerase chain reaction(RT-PCR) was used to measure the level of Fractalkine mRNA.The expressions of Fractalkine protein in every group were detected with enzyme linked immunosorbent assay (ELISA).Results In normal control group,the expression level of Fractalkine mRNA was 179.0?21.0.After exposure of IL-18 for 24 h,the level of Fractalkine mRNA was 1 220.1?185.7,which was higher than that in normal control group (t=9.646 P

Determination of carvacrol and thymol in Mosla chinensis by HPLC / 中国中药杂志

<p><b>OBJECTIVE</b>To establish a quantitative method of determination of carvacrol and thymol in Mosla chinensis.</p><p><b>METHOD</b>The sample was extracted with 95% ethanol, ODS column was used with methanol-water-acetic acid (60:40:2) as mobile phase. The detection wavelength was set at 274 nm.</p><p><b>RESULT</b>The linearities of carvacrol and thymol were respectively in the range of 0.23-2.15 microg (r = 0.9999) and 0.39-2.36 microg (r = 0.9999); the average recoveries were 99.9% (RSD 1.4%) and 98.6% (RSD 1.3%); the RSD of repeatability were 1.1% and 1.6%.</p><p><b>CONCLUSION</b>The method is reliable, and can be used for quality control of M. chinensis.</p>