Objective To investigate the efficiency and safety of transfection green fluorescent protein plasmid to mouse cornea mediated by SonoVue and ultrasound.Methods Saline,plasmid+saline,plasmid+SonoVue and liposome+plasmid were injected respectively to mouse eye anterior chamber.Then the mouse eye of plasmid+saline and plasmid+SonoVue injection group were exposed to pulse wave ultrasound under 50 Hz pulse reDetition frequence,2 W/cm2 intensity and 10 minutes duration time.Fluorescence stereomicroscope was used to observe the expression of EGFP in the eye at the 1 st day,3rd day,7th day,14th and 21 st day after injection.Two mice were taken randomly from each group and were sacrificed at the 3rd day after injection.Their eyes were enucleated and made into frozen coronal sections.And fluorescence microscopy was performed to observe the type and distribution of EGFP positive cell.Tissue damage was observed in pathological section.Results EGFP was exDressed over the ocular surface in SonoVue and ultrasound group,and it obviously higher than only ultrasound exDosure group and liposome group.The expression of EGFP was not detect ed over the ocular surface in other groups.Pathological sections were not found any difference in each group.Conclusions SonoVue and ultrasound can successfully and safely transfer gene to ocular in vivo study.

Objective To explore the effect of the simvastatin administration on the expression of connective tissue growth factor （CTGF） in fihrotic lungs of rats. Methods The cats were treated with single intratracheal instillation of bleomycin （BLM） or instillation of the same volume of normal saline （NS） as a control. The administration of simvastatin（20 mg/kg）began once a day immediately or 7 days later after intratracheal BLM instillation respectively with the same volume of NS was given as a vehicle control. The rats were killed on day 7,14 and 28 respectively. Pathological alteration of lung tissue was observed hy HE staining and Masson staining. Hydroxyproline（HYP）content in lung tissue was used to determine the severity of pulmonary fibrosis. The expression of CTGF in lung tissue was exanfined by immunohistochem- istry staining and photodeusitometry. Results Histopathological changes of pulmonary fibrosis emerged gradually after the instillation of BLM. The expression level of CTGF was increased in lungs of rats after intratracheal instillation of BLM, compared with the control. The administration of simvastatin immediately or 7 days after intratracheal instillation of BLM, attenuated the histopathological changes of bleomycin- induced puhnonary fibrosis and prevented the increased expression of CTGF in lung tissue on day 28. Conclusion The adntinistration of simvastatin, immediately or 7 days after intratracheal BLM instillation, prevented the up-regulation of CTGF in fibrotic lungs of rats, which ntight be one of the mechanisms of the anti-fihrosis of simvastatin in lungs.

Objectives To investigate the effects of Apelin 13 on myocardial metabolism in diabetic rats.Methods A total of 40 male Wister rats were randomly divided into normal control group (NC,n=8) and experimental group (n =32).Diabetic rats model were induced by high-sugar and high-fat diet combined with low-dose intraperitoneal injection of streptozotocin (STZ).The wellestablished 28 diabetic model rats were randomly divided into diabetic model group (DM,n=14) and apelin-13 treated group (n=14).In the Apelin-13 group,diabetic rats were administered Apelin-13 [0.1 μmol/(kg · d)]by intraperitoneal injection for 10 weeks,while the control group and diabetic model group were given an equal volume of 0.9％ NaCl.At the end of the 10th week,all rats were sacrificed after fasting glucose measurement.Levels of serum free fatty acids (FFA) and myocardial FFA were measured by ELISA.Expression of myocardial glucose transporter member 4 (GLUT4) were detected by immunohistochemistry.The mRNA expressions of myocardial PPARα,CD36 and CPT-1 were detected by real-time fluorescence quantitative PCR.Results Fasting blood glucose,serum FFA and myocardial FFA were significantly higher in DM group than in NC group (all P＜ 0.05).The level of plasma glucose and myocardial FFA were significant lower(P＞0.05) in Apelin-13 treated group than in DM group;but serum FFA was not significantly lower(P＜0.05).The mRNA expressions of PPARα,CD36,CPT-1 in cardiac myocyte were higher in DM group and Apelin-13 treated group than in control(P＜0.05),and lower in Apelin-13 treated group than in DM group(P＜ 0.05).The expression of myocardial GLUT4 was significantly lower in DM group(1.138±0.316)and in Apelin-13-treated group (4.631 ± 1.832) than in NC group(9.132 ± 2.156),(F=65.507,P＜ 0.05),and higher in Apelin-13-treated group than in DM group(P＜0.05).Conclusions Apelin-13 increases myocardial expression of GLUT4,improves utilization of FFA,and it can effectively reduce the expression of PPARα,CD36 and CPT-1.Therefore,it may play a vital role in the improvement of myocardial metabolism in diabetic rats.

Objective To measure the longitudinal elastic modulus in patients with acute ischemic stroke by shear wave elasotography(SWE),and to study its feasibility and related factors.Methods There were 1 79 cases with acute ischemic stroke (AIS)including 103 cases with large artery atherosclerosis and 76 cases with lacunar infarction classified according to the TOAST classification.There were 168 age and sex-matched cases as control group.Pulse wave velocity (PWV ) of bilateral carotid was measured by radiofrequency ultrasound.The 20 areas of superficial walls of bilateral carotid artery were analyzed by SWE,and the average values of longitudinal average elastic modulus (MEmean ),the maximum elastic modulus (MEmax )and minimum elastic modulus (MEmin )were calculated.The independent factors of MEmean were analyzed using multiple linear regression analysis.Results ①Compared with control group, PWV in patients with AIS was increased (P <0.05).②The MEmax and MEmean of carotid artery in patients with AIS were more than those in the control group (P <0.05).③Age,systolic blood pressure,PWV and low-density lipoprotein cholesterol(LDL-C)were positively related to MEmean (r =0.221 ,0.1 74,0.776 and 0.1 73,respectively,P <0.05 ),and were independent factors for MEmean .Conclusions SWE can evaluated the arterial stiffness using longitudinal elastic modulus of carotid.Age,systolic blood pressure,PWV and LDL were independent factors for longitudinal elastic modulus.

Suitable pretreatment of biological samples can truly reflect the role of law of the measured components played in the body and will provide experimental evidence for the studies on metabolic process, material basis of efficacy, mechanism of action, pharmacology, toxicology and the others. Biological samples include blood, urine, hair, tears, etc. There are also many samples processing methods, such as the direct protein precipitation, liquid-liquid extraction and solid phase extraction and so on. These methods could be used alone or combined.
Animals ; Body Fluids ; chemistry ; Chemical Precipitation ; Chemistry Techniques, Analytical ; methods ; Humans ; Liquid-Liquid Extraction ; Proteins ; isolation & purification ; Solid Phase Extraction

0.05).The different tilt angles impacted patients' fear psychology(?2=8.038,P=0.018).The order of the extent of children's fear psychology represented from low to high at the angle of 60?,70? and 80?,respectively.The extent of the children's fear psychology had positive correlation with the tilt angle(r=0.669,P=0.002).Conclusions The tilt test angle does not affect the positive rate of HUTT and the hemodynamics in children undergoing HUTT with angles from 60? to 80?.The children at the tilt angle 60? had less fear than at the angle of 70? or 80?.The 60? tilt angle of HUTT in children is recommended in the clinical practice.

Objective To compare the transfection rate and safety between SonoVue + ultrasound and liposome mediated gene transfection to retinal pigment epithelium (RPE) in vitro, and to approach the feasibility of SonoVue and ultrasound enhancing liposome mediated gene transfection to RPE in vitro. Methods RPE were cultured in 24 well dishes with green fluorescent protein plasmid of 2 μg/well. All the cells were divided into 5 groups: control group, ultrasound irradiance group, SonoVue + ultrasound irradiance group(2 W/cm2 and 3 W/cm2),liposome group and liposome + SonoVue + ultrasound irradiance group. There were 10 wells in every group. Gene transfection results were observed under inversion fluorescent microscope. Seventy-two hours later, gene transfer was guantified under flow cytometry. The apoptosis rates of RPE were determined with Annexin Ⅴ-FITC and PI staining by flow cytometry. Results The transfection rate of ultrasound exposure group was (1.06 ± 0.13)%. The transfection rate of SonoVue + ultrasound group was (15.81 ± 1.70)% and (20.86 ± 2.63)%. The transfection rate of liposome group was (30.06 ± 3.49)% ,and the differences between above mentioned sets were significant(P<0.05). The transfection rate of liposome + SonoVue + ultrasound was (44.51 ± 1.28) %,and the differences between above mentioned two sets were significant (P<0.05). Conclusions The transfection rate of liposome is higher than that of SonoVue + ultrasound group. SonoVue and ultrasound can obviously enhance liposome mediated EGFP gene transfection to RPE in vitro.

Salvianolic acid A (Sal A) is one of the most effective compounds isolated from the root of Salvia miltiorrhiza. Up to now, several studies regarding the pharmacokinetic profiles of Sal A have been reported, however there is no such study reported in monkeys, the species which is more similar to human. The aim of this study is to develop a LC-MS method for the determination of Sal A in monkey plasma and apply it to the pharmacokinetic studies of monkeys. After single intravenous administration of Sal A, the plasma concentration-time curves were observed and the main pharmacokinetic parameters were calculated. The plasma concentration at 2 min (C2 (min)) values were (28.343 ± 6.426), (45.679 ± 12.301) and (113.293 ± 24.360) mg x L(-1) for Rhesus monkeys treated with Sal A at 2.5, 5 and 10 mg x kg(-1). The area under the concentration-time curve (AUC(0-∞)) values were (3.316 ± 0.871), (5.754 ± 2.150) and (13.761 ± 2.825) μg x L(-1) x h, respectively. Furthermore, this method was improved and applied to the simultaneous determination of Sal A, Sal B and Sal C, which provided useful information for preclinical studies and clinical trials of Sal A, Sal B and Sal C.
Administration, Intravenous ; Animals ; Caffeic Acids ; pharmacokinetics ; Chromatography, Liquid ; Drugs, Chinese Herbal ; pharmacokinetics ; Lactates ; pharmacokinetics ; Macaca mulatta ; Mass Spectrometry ; Plant Roots ; chemistry ; Salvia miltiorrhiza ; chemistry

Objective To observe the effect of bronchial asthma combined with allergic rhinitis by azelastine hydrochloride nasal spray.Methods Eighty-seven cases of bronchial asthma combined with allergic rhinitis patients were divided into treatment group(45 cases)and control group (42 cases) by random number table method,both groups were given the treatment of bronchial asthma according to the guidelines for regulating,treatment group on the basis was combined with azelastine hydrochloride 1 spray per side,2times per day,for 3 months.Observe two groups of asthma symptoms within 3 months control situation,the number of antibiotics and hospitalization for acute exacerbation.Results Treatment group total effective rate,asthma control test score,number of antibiotics and due to the number of exacerbations hospitalization were superior to control group [93.33％(42/45) vs.76.19％(32/42),(23.80 ± 2.15)scores vs.(22.05 ±3.16) scores,13.3％ (6/45) vs.35.7％ (15/42),11.1％ (5/45) vs.33.3％ (14/42)] (P ＜ 0.05 or ＜ 0.01).Conclusion Combined by azelastine hydrochloride nasal spray may make a good control,reduce the use of antibiotics for acute attack and times of hospitalization,worthy of clinical application.

OBJECTIVETo explore the effect of Bushen Tiaojing Recipe (BTR) on the quality of oocytes, reproductive hormones, and the expression of bone morphogenetic protein-15 (BMP15) of in vitro fertilization-embryo transfer (IVF-ET) patients.

METHODSSixty infertility patients who prepared for IVF-ET were assigned to two groups according to the treatment order, the treatment group [20 cases, treated with BTR + controlled ovarian hyperstimulation (COH)] and the control group (treated with COH alone, 40 cases). Age, the time limit for infertility, basal follicle-stimulating hormone (bFSH) concentration, usage days and the dosage of gonadotropins (Gn), serum levels of estradiol (E2), luteotropic hormone (LH), and progesterone (P) on the HCG injection day, the number of retrieved occytes, the fertilization rate, the number of embryos, the high quality embryo rate, and the clinical pregnancy rate were compared. Concentrations of follicle-stimulating hormone (FSH), LH, E2, testosterone (T), and P in the follicular fluid were detected via chemiluminescence microparticle immunoassay. The mRNA and protein expression of BMP-15 in mature granulosa cells was detected by real-time fluorescent PCR and Western blot.

RESULTSThirty-two patients were pregnant and the total pregnancy rate was 53.3%. Of them, 19 were pregnant and the total pregnancy rate was 47.5% in the control group, while 20 were pregnant and the total pregnancy rate was 65.0% in the treatment group. But there was no statistical difference between the two groups (P > 0.05). Compared with the control group, the Gn dosage was lower and the high quality embryo rate was higher in the treatment group, showing statistical difference (P < 0.05). There was no statistical difference in serum concentrations of E2, LH, or P on the HCG injection day, the number of retrieved oocytes, or the fertilization rate (P > 0.05). Compared with the control group, FSH concentrations in the follicular fluid were significantly lower and LH concentrations were significantly higher in the treatment group (P < 0.05). The LH concentrations in the follicular fluid were significantly higher in pregnant patients than non-pregnant patients, showing statistical difference (P < 0.05).There was no statistical difference in E2, T, or P concentrations (P > 0.05). The mRNA and protein expression of BMP-15 in granulosa cells was higher in the treatment group than in the control group (P < 0.05). It was also higher in pregnant patients than non-pregnant patients, showing statistical difference (P < 0.05).

CONCLUSIONDuring the IVF-ET process, BTR could elevate the quality of oocytes, and increase the sensitivity of ovarian follicles to exogenous Gn, which was correlated with the mRNA and protein expression of BMP-15 in granulosa cells, and changing concentrations of FSH and LH.

Adult ; Bone Morphogenetic Protein 15 ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Embryo Transfer ; Estradiol ; blood ; metabolism ; Female ; Fertilization in Vitro ; Follicle Stimulating Hormone ; metabolism ; Follicular Fluid ; metabolism ; Humans ; Luteinizing Hormone ; blood ; metabolism ; Oocytes ; drug effects ; Pregnancy ; Progesterone ; blood ; metabolism ; Testosterone ; metabolism ; Young Adult