Objective To explore the expression of GATA-3 in pulmonary tissue of asthmatic mice and the inhibitory effect of dexamethsone(Dex)on it.Methods The Blab/c mice asthma model was induced by ovalbumin(OVA) with classic method.Twenty-four male mice were randomly divided into control group,asthmatic group and Dex treated group.The expression of GATA-3 protein was measured by immunohistochemical staining.The expression of GATA-3 mRNA was measured by reverse transcription-polymerase chain reaction(RT-PCR).The level of IL-4 in mice spleen CD4 T cell was measured by flow cytometry.The airway inflammation was evaluated by HE staining.Results The percentages of postive GATA-3,GATA-3 mRNA and IL-4 protein of asthmatic group were significantly higher than those of control group(P

Objective To investigate the transfection efficacy and expression of Ang-1 gene and proangiogenesis in vitro and vivo by ultrasound-mediated microbubble destruction.Methods 293T cells were divided into three groups:group A was given hAng-1 plasmid and microbubbles plus ultrasonic irradiation,group B was given hAng-1 plasmid and ultrasound,group C was given hAng-1 plasmid only (without ultrasound).Forty-eight hours after transfection,the transient expression rate was observed under fluorescence microscopy and flow cytometry.RT-PCR and Western blot analysis were taken to evaluate the mRNA and protein expression of Ang-1 respectively.Twenty-seven rabbit models of ligated left circumflex branch coronary artery were divided into 3 groups randomly as follow:group Ⅰ (accepted intravenous injection of SonoVue microbubble and Ang-1 plus ultrasonic irradiation),group Ⅱ (accepted intravenous injection of Ang-1 with ultrasound),group Ⅲ (control group).Myocardial contrast echocardiography (MCE) was executed on all animals before and after the treatment.Two weeks after gene delivery,RT-PCR and Western blot analysis were taken to evaluate mRNA and protein expression of Ang-1 respectively.Microvessel density (MVD) counting of infracted myocardium,observed by Factor Ⅷ immunochemical staining,was performed to value the proangiogenesis effect of Ang-1 delivered by ultrasound mediated cavitation of microbubble.Results Green fluorescence was observed in group A and B by fluorescence microscopy,which was negative in group C.The transfection expression rate was significantly improved in group A ( P ＜ 0.01).In vivo,Microbubbles could be observed in former ischemic myocardium in MCEexamination and the Ang-1 mRNA and protein could be detected in group Ⅰ.On the other hand,the contrast agent was defected obviously and none of the animals showed Ang-1 mRNA and protein expression in other two groups.The MVD counting showed significant improvement in group Ⅰ whereas other two groups didn't.ConclusionsMicrobubble-enhanced ultrasound exposure can improve the Ang-1 gene transfection expression rate observably both in vitro and in vivo.This strategy for delivering has great proangiogenesis effect in vivo.

Objective To explore the capability of Ang-1 gene delivery to acute myocardial infarction using targeted microbubbles carrying ICAM-1 antibody.Methods Thirty-seven rabbits' left circumfles branch coronary arteries were ligated for models.Three rabbits were injected with microbubbles carrying ICAM-1 to detect the ability of targeting.Thirty-four rabbit models were divided into 3 groups randomly as follow:IM group (n=12,accept direct intramuscular injection),ICAM-1 group (n=12,accept intravenous injection of targeted microbubbles and Ang-1) and control group (n=10,without any treatment).Ultrasonography were executed on all animals before and 2 weeks after the treatment.All rabbits were killed after 2 weeks and examined for Ang-1 mRNA and protein by RT-PCR and Western-Blot respectively.Microvessel density (MVD) counting of infracted myocardium,observed by factor Ⅷ immunochemical staining,was performed to value the proangiogenesis effect of Ang-1 delivered by targeted microbubbles carrying ICAM-1 antibody.The liver and the kidney in ICAM-1 group were taken to assess the systemic delivery.Results IM and ICAM-1 group showed significantly improvement in the ejection fraction (P＜0.05) while control group did not.Ang-1 mRNA and protein could be detected in IM and ICAM-1 group;however,the expression between the two groups showed no siginificant difference.None of the control animals showed Ang-1 expression.compared with ICAM-1 group,the MVD was greater in IM group.Ang-1 was not detected either in liver or in kidney in ICAM-1 group.Conclusions Targeted microbubbles carrying ICAM-1 antibody can deliver Ang-1 gene to ischemic myocardium directly.Meanwhile,it's as effective as IM injections besides the greater angiogenesis effect.This strategy improves the perfusion of acute myocardial infarction and the function of heart.

Objective To test the efficiency of gene transfer and expression mediated by ultrasound and microbubble strategy in 293T cell. Methods SonoVue microbubbles were mixed with pla.smid DNA encoding hAng-1 and green fluorescent protein. The mixture of the DNA and microbubbles was administer to cultured 293T cells by ultrasound exposue. The ultrasound condition was 1. 5 W/cm~2 and 30 s. Forty eight hours later, transfer rate was assessed by fluorescence microscopy and flow cytometry. Cell viability was assayed by Trypan Blue staining. RT-PCR and Western blot analysis was used to examine the expression of hAng-1 mRNA and protein. Agarose gel electrophoresis was used to evaluate the integrity of the plasmid. Results The transfection expression rate of eGFP in 293T cells was markedly increased with the additon of 20% microbubbles and 15 mg/L DNA. Fetal calf serum had no influence on the gene transfer rate. Ultrasound irradiation combined with microbubbles couldn't destroy the integrity of plasmid. Conclusions Ultrasound-mediated microbubbles destruction can increase the transfection and expression of hAng-1 gene in 293T.

Objective To certificate the effects on transfection ratio and cells viability of ultrasound (US) acoustic intensity, radiation duration, microbubbles concentration and DNA concentration in delivery human angiopioetin-1 gene (hAng-1) into 293T cells by SonoVue microbubbles and decide the optimal transfection parameters. Methods Mix 293T cells and SonoVue microbubbles linked with eGFP-C_3-hAng-1 in a different way, detect the gene transfection ratio and cells viability under the various US intensity, radiation duration, microbubbles and DNA concentrations. Results The gene expression would be increased if enhanced the intenstiy of US,radiation time,microbubbles and DNA concentrations,and the cells viability would be kept more than 90% ( P <0. 01). Whereas,if the US intensity increased over 1. 5 W/cm~2 ,the duration over 30 s and microbubbles and DNA concentrations over 20% and 15 mg/L respectively,the gene expression would not increase significantly ( P > 0. 05),whereas coupled with obviously decreased cells viability( P <0. 01). Conclusions The optimal conditions of deliver hAng-1 gene into 293T cells by SonoVue microbubbles was mixing cells and microbubbles in a cell wall-sticky way,US intensity was 1. 5 W/cm~2, duration 30 s,20% microbubbles and 15 mg/L DNA concentration.

Objective To study the SRSF2 mutations in acute myeloid leukemia (AML) patients by using high-resolution melting analysis (HRMA). Methods PCR-HRMA analysis was performed to screen SRSF2 mutations in 140 cases with AML, and the direct DNA sequencing was used to confirm the HRMA results. Results Five percent (7/140) of AML patients were found with heterozygous SRSF2 mutations, including one case of P95R mutation, two case of P95L mutation, and four cases of P95H mutation, the above mutations were confirmed by direct DNA sequencing. The maximal sensitivity of HRMA in detecting SRSF2 mutation was close to 10%. There were no difference in gender, age and blood parameters among cases with or without SRSF2 mutations (P > 0.05). The overall survival (OS) of patients with SRSF2 mutations was inferior to those without SRSF2 mutations in AML patients (P=0.016). Conclusions HRMA analysis was a convenient, rapid, specific, high-throughput technique for scanning of SRSF2 gene mutations in AML patients. SRSF2 mutation may predict the adverse prognosis in AML patients.

BACKGROUND: Animal experiments showed that recruitment maneuver (RM) and protective ventilation strategy of the lung could improve oxygenation and reduce extravascular lung water. This study was to investigate the effects of RM on respiratory mechanics and extravascular lung water index ( EVLWI) in patients with acute respiratory distress syndrome (ARDS). METHODS: Thirty patients with ARDS were randomized into a RM group and a non-RM group. In the RM group, after basic mechanical ventilation stabilized for 30 minutes, RM was performed and repeated once every 12 hours for 3 days. In the non-RM group, lung protective strategy was conducted without RM. Oxygenation index (PaO2/FiO2), peak inspiratory pressure (PIP), Plateau pressure (Pplat), static pulmonary compliance (Cst) and EVLWI of patients before treatment and at 12, 24, 48, 72 hours after the treatment were measured and compared between the groups. Hemodynamic changes were observed before and after RM. One-way ANOVA, Student's t test and Fisher's exact test were used to process the data. RESULTS: The levels of PaO2/FiO2 and Cst increased after treatment in the two groups, but they were higher in the RM group than in the non-RM group (P<0.05). The PIP and Pplat decreased after treatment in the two groups, but they were lower in the RM group than in the non-RM group (P<0.05). The EVLWI in the two groups showed downward trend after treatment (P<0.05), and the differences were signifcant at all time points (P<0.01); the EVLWI in the RM group was lower than that in the non-RM group at 12, 24, 48 and 72 hours (P<0.05 or P<0.01). Compared with pre-RM, hemodynamics changes during RM were significantly different (P<0.01); compared with pre-RM, the changes were not significantly different at 120 seconds after the end of RM (P>0.05). CONCLUSIONS: RM could reduce EVLWI, increase oxygenation and lung compliance. The effect of RM on hemodynamics was transient.

Abstract?AlM:To discuss the feasibility of using the visual quality analyzer KR - 1W to guide the relatively personalized aspheric intraocular lens ( lOL ) implants to make the whole eye spherical aberration close to 0. 1μm.?METHODS: ln this prospective case series study, the corneal spherical aberration with 6mm aperture of 73 patients (100 eyes) was measured with KR-1W Visual Function Analyzer 1d before surgery. For the sake of the whole postoperative spherical aberration were close to 0. 1μm, 9 cases ( 16 eyes ) with corneal spherical aberration <0. 15μm were implanted Sofport Advanced Optic lOL, named AO group;45 cases ( 57 eyes ) with corneal spherical aberration 0. 25~0. 3μm were implanted AcrySof lQ lOL, named lQ group;19 cases (27 eyes) with corneal spherical aberration > 0. 35μm were implanted Tecnis ZA9003 lOL, named Tecnis group. Aspherical lOL was implanted after phacoemulsification through a cornea 2. 75mm incision without suture. Uncorrected visual acuity, beat corrected visual acuity, spherical aberration of the whole eye and jnternal optics (mainly lOL) at 6mm pupil diameter were examined at 3mo postoperatively. The relevant data were analyzed using t-test and variance analysis.?RESULTS: The whole ocular spherical aberration at 6mm pupil diameter in all postoperative were 0. 084 ± 0. 032μm;in Tecnis group, the data were 0. 091 ± 0. 021μm;in AO group, the data were 0. 0814-0. 013μm;lQ group were0. 093 ± 0. 042μm. There was no significantly different between the predicted value and actual value of ocular spherical aberration at 6 mm pupil diameter in all postoperative ( t = 1. 932, P = 0. 061 ) and in the three groups. The difference value in the predicted values of the preoperative spherical aberrations of the whole eye and the actual values after surgery was 0. 013±0. 041μm; there was no statistically significant difference ( F=2. 537, P=0. 091 ) . Respectively compared the uncorrected visual acuity and besta corrected visual acuity among three groups of postoperative, no significant difference were found (F=0. 897, P=0. 421;F=1. 423, P=0. 097).?CONCLUSlON: Personality selection of aspheric lOL based on preoperative corneal spherical aberration of patients is feasible and produces satisfactory target postoperative total spherical aberration.

OBJECTIVETo develop a quality analysis method based on self-reference principal for dissolution determination of Shuanghuanglian capsules.

METHODDissolution of Shuanghuanglian capsules was determined by principal component analysis consociated HPLC method.

RESULTThe liner of regression equation was good. The average recovery rates of quality assurance samples (QA) and quality control samples (QC) were all no less than 96. 0%. Dissolution curves of Shuanghuanlian capsules of different manufacturers and different batches of the same manufacturer had obvious disparity.

CONCLUSIONThe method can better evaluate the dissolution conditions of Shuanghuanglian capsules. The prospect of the method is expected for assessing the dissolution of other oral solid dosage of traditional Chinese medicines.

A new flavonoid glycoside, (-)-2S-8-methyl-5,7,4'-trihydroxyflavanone-7-O-β-D-glucopyranoside (1), along with five known ones, quercetin-3-O-(2"-galloyl)-α-L-arabinoside (2), kaempferol-3-O-α-L-arabinoside (3), guaijaverin (4), trifolin (5) and hyperin (6), was isolated from the leaves of Eucalyptus robusta. Their structures with absolute configurations were elucidated by NMR, HR-ESI-MS, CD spectra data and physicochemical methods. In addition, 2-6 were isolated from E. robusta for the first time.
Eucalyptus ; chemistry ; Flavonoids ; chemistry ; isolation & purification ; Glycosides ; chemistry ; isolation & purification ; Plant Leaves ; chemistry