AIM: To generate the angiopoietin-1 recombinant adenovirus for the further studies about adenoviral mediated angiopoietin-1 gene transfer in ischemic myocardium and its effect on the development of neoangiogenesis.METHODS: Angiopoietin-1-PAxCAwt cosmid DNA and adenoviral DNA-TPC were cotransfected to 293 cells by the use of calcium phosphate precipitation method. All recombinant adenoviruses were propagated and titrated in 293 cells, purified by cesium chloride density purification, dialyzed, and stored at-70 ℃. RESULTS: The result shows that the direction of the insert in the cosmid is correct and the replication-deficient angiopoietin-1 recombinant adenovirus was generated efficiently by COS/TPC homologous recombination, with the titers of 5.6?10 11 pfu/L. The viral stocks were demonstrated to be free of replication-competent wild type adenoviruses. The virus stocks in high titer were harvested in our experiment. CONCLUSION: The COS/TPC is an efficient method to prepare recombinant adenovirus and the angiopoietin-1 recombinant adenovirus could be further used in gene therapy.

Objective To review the surgical methods and mid-term results of mitral valve repair in pediatric patients with moderate to severe mitral regurgitation (MR).Methods 132 children with moderate to severe MR,aged (18.9 ± 7.2)months,weighted(11.3 ±4.8) kg.The etiology for mitral regurgitation is congenital heart disease in 126 cases,infective endocarditis in 5 cases and Marfan syndrome in 1 case.Mitral valvuloplasty(MVP) was performed with cardiopulmonary bypass under moderate systemic hypothermia.The methods of MVP included annuloplasty,annuloplasty ring,cleft closure,reconstruction of posterior leaflet.The coucomitant cardiac anomalies were treated at the same time.The results of repair were evaluated by saline injection test and transesophageal echocardiography (TEE) during operation.Results Intra-operative TEE results: 131 cases had none to mild MR,and only one case had moderate MR.The patient underwent second repair immediately,subsequent TEE was mild.Mean cardiopulmonary bypass (CPB) time was (80.0 ± 31.1) minutes.Mean aortic clump time was (48.0 ± 17.9) minutes.The in-hospital mortality was 2.3％ (3 cases died).One died of heart failure on postoperative day 7,the other died of low cardiac output syndrome resulting on postoperative day 2.Another one was large ventricular septal defect(VSD) with pulmonary hypertension (PH),died of pulmonary infection.Mean postoperative ventilation time was (34.4 ± 31.9) hours,and mean postoperative inhosptial time was (9.0 ± 5.4) days.The average follow-up period was (40.5 ± 8.3) months (2 to 74 months).122 cases were fully followed up.Echocardiography showed that moderate MR was in 7 patients,and 3 patients had severe MR.4 patients underwent re-do mitral valve repair or mitral valve replacement.There was no late death.The overall survival rate at 5 years was 97.7％ and the overall freedom from reoperation at 5 years was 92.0％.Conclusion Pediatric patients with moderate to severe MR need early surgical treatment,the early and mid-term results were satisfactory.Individualized treatment protocol based on specific pathology was the keypoint of surgical therapy.

Taking α-asarone as model drug, mono methoxy polyethylene glycol-polylactic acid copolymer (mPEG-PLA) as the drug carrier material to prepare drug-loading nanoparticles by premix membrane emulsification for nasal administration. The prepared nanoparticles were spherical with smooth surface and average particle size of 360 nm. Polydispersity index (PDI) was 0. 030, average drug loading of (11.5 ± 0.045) % (n = 3), and the encapsulation efficiency of (86.34 ± 0.11) % (n = 3). X-ray diffraction and differential scanning calorimetry results showed that, α-asarone existed in mPEG-PLA carrier in amorphous or molecular state, different from simple physical mixture. In the in vitro release test in simulated human nasal cavity, α-asarone apis can be released quickly at close to 94% at 102 h, in line with the first-order kinetics (R² = 0.981 9). mPEG-PLA drug-loading nanoparticles release only 54%, with slow release effect, in line with Riger-Peppas model (R² = 0.967 9, n = 0.630 2), for non-fick diffusion, released by the spread of drugs and skeleton dissolution dual control. This provided the foundation for nasal drug delivery in vivo pharmacokinetic study.
Administration, Intranasal ; Anisoles ; chemistry ; Calorimetry, Differential Scanning ; Nanoparticles ; chemistry ; Polyesters ; chemistry ; Polyethylene Glycols ; chemistry ; Solubility ; X-Ray Diffraction

This study was aimed to investigate the immunophenotypic characteristics of 109 cases of B-cell chronic lymphoid leukemia (B-CLL) so as to provide evidences for the diagnosis and therapy of B-CLL, and for the detection of the minimal residual disease and its prognosis. Immunophenotyping was performed in 109 patients of B-CLL by two/three color multiparameter flow cytometry analysis using a panel of monoclonal antibodies. The results showed that in 109 cases of B-CLL, all cases expressed CD19, the positive ratios of other B lineage antigen such as CD20, CD22 and CD23 were 95.40%, 94.50%, 86.20% respectively. None of the B-CLL cases expressed CD10. The expression ratio of FMC-7 and CD38 in 105 cases of B-CLL were 28.60% and 36.20%. Among the B-CLL cases the CD5(+) cells amounted to 86.23%, CD5(-) cells amounted to 13.76%, ZAP-70 protein was expressed in 12 out of 50 patients. It is concluded that immunophenotypic data are very useful for the diagnosis and detection of minimal residual disease of B-CLL, and the relationship between the immunophenotypic characteristics and the prognosis of B-CLL needs further study.
Adult ; Aged ; Aged, 80 and over ; Female ; Flow Cytometry ; Humans ; Immunophenotyping ; Leukemia, Lymphocytic, Chronic, B-Cell ; diagnosis ; immunology ; Male ; Middle Aged ; Neoplasm, Residual ; diagnosis ; immunology ; Prognosis

Objective To summarize our clinical experience of surgical treatment for pediatric patients with ventricular septal defect(VSD) and mitral regurgitation(MR).Methods A retrospective study was performed including consecutive 84 patients with VSD and MR receiving mitral valvuloplasty(MVP) and VSD closure from January 2006 to January 2012 in Shanghai Xinhua Hospital.All patients were associated with pulmonary hypertension(PH,32-85 mm Hg).The diameters of ventricular septal defects were between 0.7 and 1.6 cm.Echocardiography showed that trivial MR (+) in 9 cases,mild MR (++)in 18 cases,moderate MR(+++) in 33 cases,and severe MR(++++) in 24 cases.VSD closure and MVP were performed with cardiopulmonary bypass under moderate systemic hypothermia.The results of repair were evaluated by transesophageal echocardiography (TEE) during operation.Results Intra-operative TEE results: no residual shunt of VSD,none MR in 80 cases,residual trivial MR in 4 cases.Mean Cardiopulmonary bypass (CPB) time was (84.6 ± 18.5) mins.Mean Aortic clump time was(50.8 ± 11.5) mins.Mean postoperative ventilation time was (38.7 ± 30.2) hours,and mean postoperative inhosptial time was(10.5 ±4.6) days.The in-hospital mortality was 1.2％ (1 case died).78 cases were fully followed up.There was no late death.Echocardiography showed that none MR in 62 cases,trivial MR in 10 cases,mild MR in 4 cases,moderate MR in 2 patients.The overall freedom from reoperation at 5 years was (97.4 ± 1.8) ％.Conclusion Ventricular septal defect with pulmonary hypertension need early surgical repair.MR was treated at the same time of VSD closure could effectively improve the surgical outcome of pediatric patients with ventricular septal defect and mitral regurgitation.

Chemical constituents in ethyl acetate and butanol fractions of ethanol extracts from Acorus tatarinowii were separated by column chromatography. Bufo skeletal muscle fatigue model was established to study the anti-fatigue activity of separated compounds. Five compounds were separated and identified by spectroscopic analysis as acoramone(1),cycloartenone(2),2,4,5-trimethoxyl-2'-butoxy-1,2-phenyl propandiol(3),5-hydroxymethyl furfural(4), and 5-butoxymethyl furfural(5). Compound 3 was a new compound, and compounds 2 and 5 were separated from this plant for the first time. Compound 4 exhibited a notable anti-fatigue activity.
Acorus ; chemistry ; Animals ; Bufonidae ; Fatigue ; drug therapy ; Muscle, Skeletal ; drug effects ; Plant Extracts ; chemistry ; pharmacology

OBJECTIVETo explore the mitochondrial toxicities induced by zidovudine (AZT) and adefovir dipivoxil (ADV) antiviral drugs using a rat model system.

METHODSTwelve healthy Sprague-Dawley rats were randomly divided into three equal groups and treated by oral gavage with zidovudine (125 mg/kg/day), adefovir (40 mg/kg/day), or saline (equal volume) for 28 days. The rats' body weights were measured once a week, and blood was collected every two weeks for blood and biochemical tests. All animals were sacrificed at the end of treatment, and liver, kidney, skeletal muscle, and cardiac muscle were collected by necropsy. Mitochondria were isolated from the respective tissue samples, and the activities of respiratory chain complexes were measured. DNA was purified from each sample and the mitochondrial DNA (mtDNA) content was monitored by quantitative real time PCR. Mitochondrial morphology was analyzed under electron microscope.

RESULTSNo significant adverse effects, including body weight loss, abnormal blood or biochemistry, were observed in rats treated with AZT or ADV. The activities of mitochondrial cytochrome c oxidase in liver and cardiac muscle were slightly decreased in rats treated with AZT (liver: 9.44+/-3.09 vs. 17.8+/-12.38, P?=?0.21; cardiac muscle: 32.74+/-5.52 vs. 24.74+/-20.59, P?=?0.28; kidney: 4.42+/-1.53 vs. 14.45+/-13.75, P?=?0.18; skeletal muscle: 33.75+/-8.74 vs. 40.04+/-2.49, P?=?0.45). The mtDNA content was significantly decreased in cardiac muscle of AZT-treated rats (cardiac muscle: 0.15+/-0.13 vs. 0.32+/-0.42, P?=?0.85). The morphology of mitochondria in liver, kidney, skeletal muscle, and cardiac muscle was significantly altered in the AZT-treated rats and included disappearance of the outer membrane, severely damaged structure, and swollen or completely absent cristae. No obvious effects were noted in the ADV- or saline-treated rats.

CONCLUSIONSignificant adverse effects related to mitochondrial toxicity were observed in rats treated with AZT. The slightly decreased mtDNA content in ADV-treated rats may suggest that this antiviral drug can also cause mitochondrial toxic effects.

Adenine ; adverse effects ; analogs & derivatives ; Animals ; DNA, Mitochondrial ; drug effects ; Electron Transport Complex IV ; metabolism ; Female ; Kidney ; enzymology ; Liver ; enzymology ; Mitochondria ; drug effects ; metabolism ; Mitochondria, Heart ; drug effects ; Mitochondria, Liver ; drug effects ; Mitochondria, Muscle ; drug effects ; Muscle, Skeletal ; enzymology ; Myocardium ; enzymology ; Organophosphonates ; adverse effects ; Rats ; Rats, Sprague-Dawley ; Zidovudine ; adverse effects

OBJECTIVETo study on effect of concentration of catalpol and 5-hydroxy methyl-2-furaldehyde (5-HMF) from Rehmanniae Radix at various processing.

METHODThe Rehmanniae Radix was dried and prepared from the steaming process with 10% ethanol, 50% ethanol at 90 degrees C and 100 degrees C each other. And the changes of catalpol and 5-HMF was determinated. The extraction of 5-HMF and catalpol was sonicated in 30% methanol for 2 h. The analysis of 5-HMF and catalpol was conducted by HPLC with reversed-phase C-18 column and detected under UV 284 nm, 204 nm. Elution was carried out at 1.0 mL min(-1) with 3% acetonitrile.

RESULTFrom this analysis, we found out that the content of catalpol was decreased with the number of processing times, and content of 5-HMF was increased with the number of processing times at various processing. The temperature and concentration of ethanol can effect on content of catalpol and 5-HMF at processing. The Cooked Rehmanniae Radix processed at 100 degrees C, 10% ethanol is best. And the content of 5-HMF processed for more than 7 times was accorded with standard of Korea phamcopoetia.

CONCLUSIONAnalyze the effect of concentration of catalpol and 5-HMF from Rehmanniae Radix at various processing, and provide the foundation for further study.

Chromatography, High Pressure Liquid ; Ethanol ; Furaldehyde ; analogs & derivatives ; analysis ; Glucosides ; analysis ; Hot Temperature ; Iridoid Glucosides ; Iridoids ; analysis ; Plant Tubers ; chemistry ; Plants, Medicinal ; chemistry ; Rehmannia ; chemistry ; Technology, Pharmaceutical ; methods

OBJECTIVETo explore the utility of chromatographic data for quality control of Paeonia suffruticosa, a Chinese herbal medicine.

METHODChromatographic fingerprints of Paeonia suffruticosa were determined by HPLC, the clustering analysis was used for data processing.

RESULTThe quantitative differences among different growing areas were found and could be used to classify herbals from different growing areas, while there seemed to be no quantitative differences for processing factor.

CONCLUSIONThe method could be used in quality control for monitoring herbal quality control, and the result also suggested that different growing areas may greatly affected the herbal qualities.

China ; Chromatography, High Pressure Liquid ; instrumentation ; methods ; Cluster Analysis ; Drugs, Chinese Herbal ; analysis ; chemistry ; standards ; Ecosystem ; Paeonia ; chemistry ; growth & development ; Plant Roots ; chemistry ; growth & development ; Plants, Medicinal ; chemistry ; growth & development ; Quality Control ; Reproducibility of Results