Aim: A new derivative LC-MS method was developed for the simultaneous determination of zofenopril and its active metabolite zofenoprilat to investigate the pharmacokinetic characteristics of zofenopril and zofenopri-lat in healthy Chinese volunteers after single and multiple oral doses of zofenopril calcium tablets. Methods: Ten Chinese healthy volunteers were given three single oral doses of 15,30, and 60 mg, respectively, and consecutively the multiple doses of 30 mg. The concentration and pharmacokinetic parameters of both the parent drug and the active metabolite were simultaneously determined by derivative LC-MS method using p-bromophenacyl bromide (p-BPB) as the derivative reagent. Results: After the single oral administrations of 15, 30, and 60 mg of zofeno-pril calcium, there was no significant difference in the t_(1/2) of both zofenopril and zofenoprilat among the three do-ses. The values of AUC_(0-24h) and c_(max) for both zofenopril and zofenoprilat showed the good linearities to the dosage over the dose range from 15 mg to 60 mg. There were no significant differences in AUC_(0-24h) and c_(max) for both com-pounds between female and male volunteers. After multiple oral administration( 30 mg once daily for 6 days ), the average steady state plasma concentration( c_(av)) for zofenopril was (5. 07 ±1. 06) ng/mL with the degree of fluctu-ation (DF) of 14. 26 ± 2. 94. The c_(av) for zofenoprilat was (6. 28 ± 1. 87) ng/mL with the DF of 11. 61 ±4. 68. The accumulation index values for zofenopril and zofenoprilat were 0. 94 ± 0. 31 and 0. 83±0. 13, respec-tively. Conclusion: Both zofenopril and zofenoprilat were demonstrated of linear kinetics after single administra-tion and showed no accumulation after multiple administration of the test zofenopril calcium tablets. There was significant difference in the pharmacokinetic characteristics for zofenopril calcium between healthy Chinese and European volunteers.

The 1.23 kb DNA fragment encoding the early protein EP0 of pseudorabies virus (PRV) Ea strain was amplified by PCR technique and cloned into pBluescriptII sk+.Three sequencing plasmids containing the partial fragment of the EP0 gene were constructed and the sequences were obtained by Sanger's sequencing technique. Compared with PRV InFh strain, there were multipile site-mutations and a deleted-mutation in the EP0 gene of PRV strain Ea，and the diversity of amino acid residues also existed.Then, the EP0 gene was inserted into an expression vector, pET-28a, fused into the downstream of the 6ΧHis-Tag in frame, to yield the expression plasmid pETEP0. After induction by IPTG, a high expression of fusion protein was obtained, SDS-PAGE analysis and Western blotting showed that the fusion protein was 62kD and the protein was specific to antisera against PRV Ea strain. This indicated that the EP0 gene be expressed in BL21(DE3) and the expression products have immuno-genicity.

Objective:To observe the efficacy and safety of dexzopiclone plus auricular acupressure in intervening primary insomnia.Methods:A total of 72 participants who met the inclusion criteria were enrolled in a randomized controlled trial,with 36 cases allocated to a treatment group and 36 cases allocated to a control group.Both groups were given dexzopiclone as the routine treatment.Patients in the treatment group were given auricular acupressure with Wang Bu Liu Xing (Semen Vaccariae) seeds at the auricular acupoints related to sleep and emotion based on meridian theory,whereas for patients in the control group,the medical plasters with Wang Bu Liu Xing (Semen Vaccariae) seeds were only gently stuck to acupoints unrelated to sleep without stimulation.Patients in both groups were required to visit the hospital once a week for replacing the seeds and plasters.The course of intervention lasted for 8 weeks and the patients were followed up for another 4 weeks.Pittsburgh sleep quality index (PSQI) and Karolinska sleep diary (KSD) were used to evaluate the outcomes.Meanwhile,adverse effects were monitored and recorded.Results:In the enrolled 72 cases,4 patients (one in the treatment group and three in the control group) reported thirst and a bitter taste,and one case in the control group reported nausea and vomiting.At last,3 cases in the control group dropped out for adverse reactions,and 69 cases completed the clinical trial.After 8 weeks of treatment,the global scores of PSQI in both treatment and control groups decreased significantly compared with the baseline (both P＜0.001).Furthermore,the global score of PSQI in the treatment group was lower than that in the control group (P＜0.01).The global scores of PSQI in both groups at the follow-up were significantly different from the baseline (both P＜0.001),but insignificantly different compared with the post-treatment results (both P＞0.05).According to KSD,both treatment protocols could prolong the total sleep time,shorten sleep-onset latency,improve sleep efficacy and sleep quality significantly,and the changes in the treatment group were more significant.The total effective rate was 88.9％ in the treatment group,higher than 81.8％ in the control group,though the difference was statistically insignificant (P＞0.05).Conclusion:Dexzopiclone plus auricular acupressure is effective and safe for patients with primary insomnia both in short and long terms,and it is more effective than monotherapy of dexzopiclone.

Objective This study is aimed at determining whether anti-von Willebrand factor (VWF) autoantibodies are present in the plasma of idiopathic thrombotic thrombocytopenic purpura (TTP) patients with normal ADAMTS13 activity and undetectable anti-ADAMTS13 antibodies,and at examining whether murine monoclonal antibodies (mAbs) against human VWF decrease the susceptibility of VWF to ADAMTS13 in vitro.Methods Anti-VWF autoantibodies and ultralarge VWF (UL-VWF) multimers were measured in plasma samples of 53 adult patients with idiopathic TTP by enzyme-linked immunosorbent assay and sodium dodecylsulphate-agarose gel electrophoresis,respectively.Moreover,the effects of eight murine mAbs to different human VWF domains on VWF cleavage by ADAMTS13 were evaluated under fluid shear stress and static/denaturing conditions,respectively.Results Anti-VWF antibodies and UL-VWF multimers were detected in two TTP patients with normal ADAMTS13 activity and undetectable anti-ADAMTS13antibodies.The SZ34,an anti-VWF mAb,inhibited VWF proteolysis mediated by ADAMTS13 under flow,but not static conditions.Conclusion Anti-VWF antibody may be one of the causes of idiopathic TTP with normal ADAMTS13 activity and undetectable anti-ADAMTS13 antibodies.

OBJECTIVERegional left ventricular (LV) function could be detected by measuring peak-systolic strain by speckle tracking imaging (STI). We evaluated the value of STI combined with adenosine stress echocardiography on assessing myocardial viability in patients with myocardial infarction (MI).

METHODSTwo dimensional echocardiography was performed at rest and after adenosine stress echocardiography (infused at 140 µg×kg(-1)×min(-1) over a period of 6 min) in 39 stable patients with previous MI. Peak-systolic (Speak-sys) circumferential strain, radial strain and longitudinal strain were assessed by STI. Radionuclide myocardial perfusion/metabolic imaging served as the "gold standard" to detection of myocardial viability.

RESULTS(1) There were 215 viable and 153 non-viable regions among 368 abnormal motion segments out of 624 segments in 39 MI patients according to radionuclide imaging results. (2) Speak-sys was similar between viable and nonviable myocardium at rest (all P > 0.05). After adenosine infusion, radial Speak-sys [(37.98 ± 5.45)% vs. (30.22 ± 5.47)%], longitudinal Speak-sys [(-23.71 ± 4.53)% vs. (-17.52 ± 4.34)%] increased significantly (P < 0.05)in viable segments compared to baseline levels and were significantly higher than in nonviable segments radial Speak-sys [(37.98 ± 5.45)% vs. (30.12 ± 5.37)%] and longitudinal Speak-sys [(-23.71 ± 4.53)% vs. (-16.95 ± 4.62)%] (P < 0.05), while remained unchanged in nonviable segments before and after adenosine infusion. Circumferential Speak-sys was similar before and after adenosine infusion in both viable and nonviable segments (all P > 0.05). (3) Delta radial strain change > 9.8% has a sensitivity of 82.3% and a specificity of 81.1% whereas a delta change of longitudinal strain > 16.5% has a sensitivity of 83.5% and a specificity of 92.3% for detecting viable segments.

CONCLUSIONSSpeckle tracking imaging combined with adenosine stress echocardiography could serve as a new and reliable method of assessing myocardial viability.

Aged ; Aged, 80 and over ; Cell Survival ; Echocardiography ; Female ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; diagnostic imaging ; Myocardium ; cytology ; Radionuclide Imaging

This study was aimed to investigate the effect of CD44 gene silence on the drug resistance and biologic activity of human multidrug resistant leukemia cell line K562/A02. The oligonucleotides of CD44 gene were designed according to related data of GenBank, double-stranded DNA was produced by annealing, and was inserted into pGCsilencerU6/Neo/GFP vector. The resultant recombinant plasmid pGCsiRNA-CD44 was transfected into K562/A02 cell line. Expressions of CD44, mdr-1 and blc-2 mRNA were assayed by real time RT-PCR. The 50% inhibitory concentration (IC50) of doxorubicin (ADM) for K562/A02 cell line was determined by MTT method. Cell cycle was determined by flow cytometry. The morphology of apoptotic cells was examined by Hochst 33258 staining. The results indicated that the siRNA eukaryotic plasmid directing at CD44 gene could effectively silence the CD44 gene of K562/A02 cells; as compared with control group, the CD44 expression in K562/A02 cells transfected with 4 pGCsiRNA-CD44 plasmids was obviously inhibited, while the inhibition of CD44 expression in cells transfected with siCD44-1 was strongest. After being transfected with pGCsiRNA-CD44, the expression of CD44 mRNA in K562/A02 cells reduced by 64.1% (p<0.05), at the same time the expression of mdr-1 and bcl-2 mRNA in pGCsiRNA-CD44-transfected K562/A02 cells reduced by 25.6% and 50.8% respectively. IC50 of K562/A02 cells after transfection decreased to (8.77+/-1.63) microg/ml and was obviously lower than that of control (17.97+/-1.61) microg/ml (p<0.01). After transfection for 48 hours, the ratio of K562/A02 cells in G0/G1 increased by 10.7%, and the cells displayed karyopyknosis, nuclear margination and apoptotic bodies. It is concluded that the siRNA plasmid specifically targeting CD44 gene can remarkably down-regulate the expression of CD44 gene, inhibit K562/A02 cell proliferation, induce its apoptosis and effectively reverse the multidrug resistance of K562/A02 cells.
Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Gene Silencing ; Genetic Vectors ; Humans ; Hyaluronan Receptors ; genetics ; K562 Cells ; RNA, Small Interfering

Adolescent ; Amino Acid Metabolism, Inborn Errors ; complications ; Child ; Female ; Humans ; Male ; Mental Disorders ; etiology

OBJECTIVETo investigate the risk factors in postoperative cognitive dysfunction (POCD) induced by patient self-controlled intravenous analgesia (PCIA).

METHODSA case-control study included 103 patients with POCD(P group), assessed by Mini-Mental-State test and 103 cognitive normal controls (NP group). The cases and controls were matched for age, gender,history of operation and PCIA volume dose. The relationship between POCD and various factors was analyzed by univariate and multivariate analysis. Spss 11.5 of statistical software was used for data analysis.

RESULTSData from univariate analysis showed that the history of cerebral trauma, education level and VAS score had significant differences between P group and NP group. Multivariate analysis conformed that the history of cerebral trauma, VAS score and education level were significantly related to POCD induced by PCIA and their ORs (95% CI) were 4.261 (1.344-13.508), 2.364 (1.209-4.624), 0.312 (0.170-0.573) respectively.

CONCLUSIONPatient's history of cerebral trauma and low VAS score were independent risk factors of POCD induced by PCIA and high education level seemed to be a protective factor.

Aged ; Analgesia, Patient-Controlled ; adverse effects ; Case-Control Studies ; Cognition Disorders ; etiology ; Female ; Humans ; Male ; Middle Aged ; Postoperative Complications ; etiology ; Risk Factors

Objective To explore the diagnostic and score value of ultrasound on hemophiliac arthropathy referring to MRI on the diagnosis and score of hemophiliac arthropathy Methods The ultrasound and MRI examinations were performed on 42 joints of 42 hemophilia patients 14 knees 14 ankles and 14 elbows The consistency of ultrasound and magnetic resonance imaging in the detection and score of joint diseases was compared Finally inter-and intra-observer agreement of ultrasound scoring system were tested Results The consistency of ultrasound and magnetic resonance imaging was excellent κ=0 763-0 896 P < 0 001 in the detection of early soft tissue lesions effusion or hemarthrosis synovial hypertrophy hemosiderin excellent κ=0 793 P <0 001 in the detection of cartilage loss poor κ=0 133 P = 0 132 in the detection of erosions and poor κ= 0 100 P = 0 137 in the detection of subchondral cysts The consistency of ultrasound and magnetic resonance imaging was good to excellentκ=0 684-0 833 P < 0 001 in the score of early soft tissue lesions effusion or hemarthrosis synovial hypertrophy and hemosiderin and poor to good κ=0 145 -0 635 P <0 001 in the score of advanced osteochondral lesions cartilage loss and bone erosions The inter-observer agreement was good to excellent κ=0 676-0 870 P <0 001 for early soft tissue lesions and moderate to excellent κ=0 421- 0 75 1 P < 0 001 for advanced osteochondral lesions The intra-observer agreement was good to excellent κ=0 705-0 885 P <0 001 for early soft tissue lesions and moderate to good κ=0 532 -0 732 P <0 001 for advanced osteochondral lesions Conclusions Ultrasound plays an important role in detecting early soft tissue changes effusion or hemarthrosis synovial hypertrophy hemosiderin and cartilage loss which helps follow-up and guide clinical treatment.

OBJECTIVETo investigate the effect of PD98059 on the proliferation and cell cycle of rat hepatic stellate cells (HSCs) stimulated by acetaldehyde and explore its mechanism.

METHODSRat HSCs stimulated by acetaldehyde were incubated with different concentrations of PD98059. Cell proliferation was assessed by MTT colorimetric assay. Cell cycle was analysed by flow cytometry. The mRNA of cyclin D1 and CDK4 were examined by RT-PCR.

RESULTS20, 50, 100 micromol/L PD98059 could significantly inhibit the proliferation of HSCs stimulated by acetaldehyde in a does-dependent manner (0.109+/-0.020, 0.081+/-0.010 and 0.056+/-0.020 vs 0.146+/-0.030, F=31.385, P<0.05) and provoke G0/G1 phase arrest of HSCs stimulated by acetaldehyde in a does-dependent manner (61.9%+/-6.3%, 64.1%+/-3.3% and 70.9%+/-4.8% vs 55.2%+/-4.4%, F=16.402, P<0.05). 50, 100 micromol/L PD98059 could markedly inhibit cyclin D1 mRNA expression of HSC stimulated by acetaldehyde (0.56+/-0.04 and 0.46+/-0.03 vs 0.65+/-0.07, F=68.758, P<0.05) and CDK4 mRNA expression (0.39+/-0.07 and 0.33+/-0.05 vs 0.50+/-0.06, F=29.406, P<0.05).

CONCLUSIONThe Erk signal transduction pathway plays an important role in regulating the proliferation and cell cycle of rat hepatic stellate cells stimulated by acetaldehyde, which may be partly related to its regulative effect on the expression of cyclin D1 gene and CDK4 gene

Acetaldehyde ; pharmacology ; Animals ; Cells, Cultured ; Cyclin D1 ; metabolism ; Cyclin-Dependent Kinase 4 ; Cyclin-Dependent Kinases ; metabolism ; Enzyme Inhibitors ; pharmacology ; Flavonoids ; pharmacology ; Hepatocytes ; drug effects ; Proto-Oncogene Proteins ; Rats