This study aimed to examine the preparation of cationic lipid microbubble (CLM), and evaluate its physical and chemical properties and toxicity, measure the gene transfection efficiency by ultrasound triggered microbobble destruction (UTMD) in combination with CLM. The CLM was prepared by the method of the thin film hydration, and its morphology was observed under the electron microscopy at 1st, 3rd, 7th, 10th, and 14th day after preparation, respectively. The size, Zeta potential and stability of CLM were tested. The acute toxicity of CLM was assessed. The green fluorescent protein gene (EGFP) transfection efficiency was evaluated. The experiment grouping was as follows: naked plasmid group (P group), ultrasonic irradiation plus naked plasmid group (P-US group), naked plasmid plus CLM group (P-CLM group), naked plasmid plus ultrasound and CLM group (UTMD group). The expression of EGFP was detected by fluorescent microscopy and flow cytometry. The results showed that CLMs were spherical in shape, with the similar size and good distribution degree under the light and electron microscopies. The size of CLMs was varied from 250.4±88.3 to 399.0±99.8 nm and the Zeta potential of CLMs from 18.80±4.97 to 20.1±3.1 mV. The EGFP expression was the strongest in the UTMD group, followed by the P-CLM group, P-US group and P group. Flow cytometry results were consistent with those of fluorescent microscopy. The transfection efficiency was substantially increased in the P-US group, P-CLM group and UTMD group as compared with that in the P group, almost 7 times, 10 times and 30 times higher than that in the P group respectively. It is suggested that CLMs prepared by the method of thin film hydration are uniform in diameter, and proved non-toxic. UTMD combined with CLM can significantly increase the transfection efficiency of EGFP to targeted cells.

Objective Color doppler ultrasonography of chronic Keshan disease (CKD) was evaluated to provide evidences for clinic diagnosis of the disease. Methods From September to Novermber 2009, according to "Diagnostic criteria of Keshan disease" (GB 17021-1997), 64 cases of CKD were randomly sampled from five Keshan diseased districts in Shandong province, Zoucheng, Sishui, Yishui, Wulian, Jvxian, and Pingyi as patient group. Thirty four healthy volunteers being checked up by Shandong Institute for Endemic Diseases Control and Research were put in control group. All the subjects were examined with Color doppler ultrasonography. The indexes of cardiac structure, left ventricular (LV) systolic function and LV diastolic function were measured.Results Left atrial internal diameter, LV end-diastolic internal diameter, LV end-systolic internal diameter, right ventricular diameter, aorta diameter, right atrial transverse diameter, right atrial long diameter and left ventricle mass of the patient group[(35.38 ± 6.89), (61.57 ± 8.61), (45.39 ± 10.29), (17.22 ± 3.79), (28.69 ± 2.81),(38.00 ± 6.05), (42.68 ± 8.65)mm, (283.22 ± 103.12)g] were higher than that of control group[(26.70 ± 3.27),(45.41 ± 4.93), (26.91 ± 4.35), (13.76 ± 2.27), (24.09 ± 2.89), (31.50 ± 3.32), (35.82 ± 3.14) mm, (156.03 ±39.86)g, t = 6.93, 10.09, 9.98, 4.87, 7.64, 5.81, 4.46, 6.90, all P＜ 0.05]. The LV ejection fraction and fractional shortening of the left ventricular of the patient group[(49.25 ± 14.33)%, (26.11 ± 9.17)%] were lower than that of control group[(73.88 ± 4.04)%, (42.88 ± 3.62)%, t = - 9.79, - 10.22, all P＜ 0.05]. Diffuse hypokinetic motion of the left ventricle reduced in 95% (61/64) of CKD patients, and 5% (3/64) of CKD patients had segmental LV dyskinesia. Seventy five per sent(48/64) of the patients accompanied with mitral regurgitation, and 39% (26/64) of these cases accompanied with tricuspid regurgitation. Meaningful Mitral or tricuspid regurgitation was not found out in control group. Conclusions The CKD patients' bore of atrio-ventricular cavity and LV mass are enlarged, and their motion of ventricle is reduced or partly reduced. They have poor heart function. Mitral regurgitation are more than tricuspid regurgitation. Color doppler Ultrasonography is important in diagnosis of chronic Keshan discase.

Objective To study the pharmacokinetics profiles of ibupro-fen and pseudo ephedrine with chlorpheniramine suspension after single and multiple dosing in healthy volunteers.Methods Three single and one multiple oral doses of ibuprofen and pseudo ephedrine with chlorphe-niramine suspension were given to healthy volunteers respectively.Ibu-profen concentrations in plasma were determined by HPLC-UV method.Pseudo ephedrine and chlorpheniramine concentrations in plasma were determined by HPLC-MS-MS method.The pharmacokinetic parameters were obtained with statistical analysis by DAS Ver 2.0.Results The main pharmacokinetic parameters of 3 single doses(ibuprofen:200,400,600 mg pseudo ephedrine:30,60,90 mg;chlorpheniramine:2,4,6 mg)and multiple dose(ibuprofe,pseudo ephedrine and chlorphe-niramine 200,30,2 mg,respectively)shown that the concentration-time curves of ibuprofen,pseudo ephedrine and chlorpheniramine were described by one-compartment open model and physiological disposi-tions were assumed by linear kinetics characteristics.Conclusion In multiple dosing study,physiological dispositions of ibuprofen,pseudo e-phedrine and chlorpheniramine all existed no induction and inhibition of drug enzyme phenomenon.

BACKGROUNDTbx1 is the major candidate gene for DiGeorge syndrome (DGS). Similar to defects observed in DGS patients, the structures disrupted in Tbx1(-/-) animal models are derived from the neural crest cells during development. Although the morphological phenotypes of some Tbx1 knock-down animal models have been well described, analysis of the cardiac performance is limited. Therefore, myocardial performance was explored in Tbx1 morpholino injected zebrafish embryos.

METHODSTo elucidate these issues, Tbx1 specific morpholino was used to reduce the function of Tbx1 in zebrafish. The differentiation of the myocardial cells was observed using whole mount in situ hybridization. Heart rates were observed and recorded under the microscope from 24 to 72 hours post fertilization (hpf). The cardiac performance was analyzed by measuring ventricular shortening fraction and atrial shortening fraction.

RESULTSTbx1 morpholino injected embryos were characterized by defects in the pharyngeal arches, otic vesicle, aortic arches and thymus. In addition, Tbx1 knock down reduced the amount of pharyngeal neural crest cells in zebrafish. Abnormal cardiac morphology was visible in nearly 20% of the Tbx1 morpholino injected embryos. The hearts in these embryos did not loop or loop incompletely. Importantly, cardiac performance and heart rate were reduced in Tbx1 morpholino injected embryos.

CONCLUSIONSTbx1 might play an essential role in the development of pharyngeal neural crest cells in zebrafish. Cardiac performance is impaired by Tbx1 knock down in zebrafish.

Animals ; Branchial Region ; cytology ; drug effects ; Heart ; drug effects ; physiology ; Heart Rate ; drug effects ; In Situ Hybridization ; Myocardium ; cytology ; Neural Crest ; cytology ; drug effects ; Oligonucleotides, Antisense ; pharmacology ; T-Box Domain Proteins ; antagonists & inhibitors ; metabolism ; Thymus Gland ; cytology ; drug effects ; Zebrafish ; embryology ; metabolism ; Zebrafish Proteins ; antagonists & inhibitors ; metabolism

OBJECTIVETo develop the techniques of total hip arthroplasty(THA) for Crowe type IV developmental dysplasia of the hip (DDH) with S-ROM prosthesis,and to assess its clinical results.

METHODSFrom October 2000 to October 2011,30 patients (36 hips) with Crowe type IV DDH underwent THA,including 6 patients with bilateral hip involved and 24 patients with unilateral. S-ROM prosthesis was adopted together with subtrochanteric transverse osteotomy. All the cementless acetabular cups were placed at the original anatomic location. The threaded cups were put in or near the level of the true acetabulum in all patients. Full coating stems were used in femoral side. All the patients were evaluated by using the Modified Harris Hip Score. Radiographic evaluations were made preoperatively and during follow-up.

RESULTSTwo patients lost of follow-up. Twenty-seven patients with 32 hips were followed up,and the average duration was 48 months (ranging from 7 to 84 months). There was 1 patient with bilateral THA died from hemorrhagic shock. Two patients could walk freely with the visible fracture lines at 12th and 18th months postoperatively. There were no complications such as infection or nerve injuries. Modified Harris Hip Score improved from preoperative 41.7+/-3.7 to postoperative 89.1+/-2.9. There was no acetabular or femoral component revision because of mal-position or loosening of the prostheses in all patients. Postoperative X-ray showed that all the prostheses in place,good integration between acetabular cups,femoral prosthesis and host bone without loosening. All bone grafts were integrated. All the hips acquired union of osteotomy and bone in-growth. None of the patients had radiographic evidence of aseptic loosening of prosthesis.

CONCLUSIONFor the complex DDH, follow methods should be used to improve therapeutic effects:good exposure of the true acetabulum,deepen acetabulum, femoral shortening, oblique osteotomy, using the S-ROM prosthesis.

Adult ; Aged ; Arthroplasty, Replacement, Hip ; methods ; Female ; Hip Dislocation, Congenital ; surgery ; Hip Prosthesis ; Humans ; Male ; Middle Aged

OBJECTIVETo study age and educational level and their relationship with fall-related injuries in Shanghai and to analyze the relevant costs.

METHODSMultistage cluster sampling was used for the selection of participants and standardized questionnaires were used for the information collection in 2006. Information on cases and deaths caused by fall-related injuries were obtained from 494 hospitals as well as from the mortality registry systems from 2001 till 2010.

RESULTSOf 45 857 participates, 674 suffered from fall-related injuries with the largest proportion among all injuries. The fall-related mortality increased from 10.63 per 100 000 in 2001 to 14.11 per 100 000 in 2010. The under-five mortality rate was the highest among children aged 0-14 years. Mortality increased dramatically among those aged 55 or above for the female and aged 60 or older for the male. Individuals with an educational level under the primary school were more likely to suffer fall-related injuries, accounting for 72.66% of all deaths and 49.24% of nonfatal cases respectively. The annual burden of fall-related injuries equated to 25.90% of the share of GDP for the healthcare, social security and welfare industries in 2006.

CONCLUSIONFall-related injuries were inversely related to victims' educational level. Children under the age of 5, women over 55 years old and men over 60 years old with an educational level lower than the primary school are the most risky groups of populations for intervention measures.

Accidental Falls ; mortality ; statistics & numerical data ; Adolescent ; Adult ; Age Distribution ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; China ; Educational Status ; Female ; Humans ; Male ; Middle Aged ; Sex Distribution ; Young Adult

OBJECTIVETo study the effect of methotrexate (MTX), a folic acid antagonist which can lead to folic acid deficient, on the cardiac development and on the expressions of BMP2b and HAS2 in zebrafish.

METHODSThe zebrafish embryos at 6-48 hrs post fertilization (hpf) were treated with various concentrations of MTX (0.5 x 10(-3), 1.0 x 10(-3) and 2.0 x 10(-3) M). At 48 hpf, the percentage of cardiac malformation and heart rate were recorded. The zebrafish embryos at 6-10 hpf treated with 1.5 x 10(-3) M MTX were used as the MTX treatment group. At 24 and 48 hpf the cardiac morphology was observed under a microscope. The expressions of BMP2b and HAS2 in zebrafish were detected by in situ antisense RNA hybridization and real-time PCR.

RESULTS6-12 hpf, the early embryonic developmental stage, was a sensitive period that MTX affected cardiac formation of zebrafish. The retardant cardiac development and the evidently abnormal cardiac morphology was found in the MTX treatment group. The results of in situ antisense RNA hybridization showed that the expressions of BMP2b and HAS2 in the zebrafish heart were reduced in the MTX treatment group at 36 and 48 hpf. The real-time PCR results demonstrated that the BMP2b expression decreased at 12, 24, 36 and 48 hpf, and that the HAS2 expression decreased at 24, 36 and 48 hpf in the treatment group compared with the control group without MTX treatment.

CONCLUSIONSThe inhibition of folic acid function may affect cardiac development of early embryos, resulting in a retardant development and a morphological abnormality of the heart in zebrafish, possibly by down-regulating the expressions of BMP2b and HAS2.

Abnormalities, Drug-Induced ; etiology ; Animals ; Bone Morphogenetic Protein 2 ; Bone Morphogenetic Proteins ; genetics ; Down-Regulation ; Folic Acid Antagonists ; toxicity ; Gene Expression Regulation ; drug effects ; Glucuronosyltransferase ; genetics ; Heart Defects, Congenital ; chemically induced ; Hyaluronan Synthases ; Methotrexate ; toxicity ; Polymerase Chain Reaction ; Zebrafish ; Zebrafish Proteins ; genetics

OBJECTIVETo determine the efficacy of phosphocreatine kinase in the early diagnosis of compartment syndrome.

METHODSForty patients with compartment syndrome of limbs were reviewed from 2005 to 2008 including 34 males and 6 females with an average age of (37.03 +/- 13.02) years. Monitoring phosphocreatine kinase continuously and dynamically after injured 2, 24 hours, 1, 2, 3 and 4 weeks later. The concentration of CK were measured by using Japanese Olympus automatic biochemistry analysator. The muscle preparations from affected extremity were taken after operation and 1, 2, 3 weeks later for biopsy.

RESULTSTwo hours later after injury, the contents of CK increased sharply and the contents of CK were about 20 times more than the nomal. Twenty-four hours later, the contents of CK reached its maximum,the contents of CK were about 42 times more than the nomal. One week later, the contents of CK recovered to normal level. Pathological changes of muscle were irreversible.

CONCLUSIONThe change of the contents of CK can reflect the progression of disease objectively. If it increased sharply, the chance of compartment syndrome was high. Monitored it dynamicly and continuously can provide assistant for early diagnosis of compartment syndrome and evaluating pathogenetic condition.

Adolescent ; Adult ; Aged ; Child ; Compartment Syndromes ; blood ; diagnosis ; Creatine Kinase ; blood ; Female ; Humans ; Male ; Middle Aged ; Time Factors

OBJECTIVETo examine the effect of particulate matter (PM) less than 10 microns in diameter (PM10) and ozone (O3) on daily mortality in Shanghai, China.

METHODSA generalized additive model with penalized spline function was used to observe the acute effect of PM10 and O3 on daily mortality.

RESULTSHigher PM10 significantly increased the effect of O3 on total mortality, and O3 also increased the effect of PM10 although the estimated increment was statistically insignificant.

CONCLUSIONOur findings provide further evidence for the effect of PM10 and O3 on daily mortality.

Air Pollutants ; toxicity ; China ; epidemiology ; Humans ; Mortality ; Ozone ; toxicity ; Urban Population

Glutamate dehydrogenase (GDH) is a key enzyme in the biosynthesis of glutamate. The GDHs from Corynebacterium glutamicum S9114 the most commonly used strain in glutamate fermentation, were purified and their molecular structures and properties characterized. The coenzymes were also studied in the hope to increase glutamate production. Cells were harvested at mid-exponential phase by centrifugation and washed with Tris-HCl buffer containing DTT and EDTA (pH 7.5). The cells were then disrupted using a French pressure cell press and the supernatant was collected by centrifugation. The extract was concentrated by 70-fold using the AKTA-100 FPLC system employing a DEAE-cellulose ion exchange column, a hydrophobic interaction chromatography (HIC) and Sephadex G-200 gel filtration. The purified extracts contained NADPH-dependent GDH and NADH-dependent GDH. Both of the enzymes were highly specific for the coenzymes. The molecular masses of the NADPH-dependent GDH and its subunit were 188kD and 32kD respectively, suggesting the enzyme is a homo-hexamer. Our data reported for the first time the presence of NADH- dependent GDH in Corynebacterium glutamicum S9114, similar to other microorganisms containing both GDHs. The NADPH-dependent and NADH-dependent GDH in Corynebacterium glutamicum S9114 may participate in the assimilation and dissimilation of ammonia respectively. The absorptions of NADPH-dependent GDH was very weak at 280nm but very high at 215nm, suggesting a low phenylalanine and tyrosine content in the enzyme.
Chromatography, Gel ; Chromatography, Ion Exchange ; Corynebacterium glutamicum ; enzymology ; Glutamate Dehydrogenase ; isolation & purification ; metabolism ; Molecular Weight ; NADP ; metabolism ; Substrate Specificity