OBJECTIVE:To study the pharmacokinetics of compound amlodipine besylate/atorvastatin calcium tablets(CABACT) following oral administration of single dose vs. multiple doses in healthy volunteers. METHODS:10 volunteers were administered a single dose of Compound amlodipine besylate/atorvastatin calcium tablets(10 mg,p.o.)or multiple doses(10 mg?d-1,p.o.) for 7 days,respectively. Plasma concentrations of amlodipine besylate and atorvastatin calcium were determined by LC-MS/MS and the pharmacokinetic parameters were calculated using DAS software. RESULTS:In single-dose study the pharmacokinetic parameters of amlodipine besylate vs. atorvastatin calcium were as follows:t1/2?(53.4?12.9)h vs.(15.4?4.6)h; Cmax(6.7?1.8)?g?L-1 vs.(18.5?4.4)?g?L-1; AUC0～120 (298.8?97.1)?g?h?L-1 vs. (118.3?48.9)?g?h?L-1; AUC0～∞(412.2?131.5) ?g?h?L-1 vs. (120.0?55.1)?g?h?L-1. In multiple-dose study the pharmacokinetic parameters were as follows:t1/2(49.5?10.3)h vs. (14.4?5.3)h; Cmax(8.7?2.5)?g?L-1 vs. (20.3?5.8)?g?L-1; AUC0～120(451.2?127.1)?g?h?L-1 vs. (136.3?54.9)?g?h?L-1; AUC0～∞(569.3?165.8) ?g?h?L-1 vs. (139.0?61.3)?g?h?L-1; Cav(2.7?0.6)、(8.3?1.3)?g?L-1; R(1.7?0.4) vs. (1.1?0.2). CONCLUSION:The elimination rates of amlodipine besylate and atorvastatin calcium do not change after oral administration of multiple doses of CABACT while slight accumulation of amlodipine besylate is founded.

Objective To investigate the relation between the expression of vascular endothelial growth factor (VEGF) and type Ⅳ collagen and the renal lesion of diabetes mellitus in diabetic rats. Methods Wistar rats were divided at random into normal control group(NC group) and diabetic model group(DM group), each group containing 20 animals. The rats models of diabetic mellitus were induced by streptozotocin(STZ). At the 8th week after induction, the expression levels of VEGF and type IV collagen in the kidney tissues in the two groups of rats were detected using immunohistochemistry. At the same time, the renal function of the rats was measured. Results At the 8th week,the expression levels of VEGF and type IV collagen of kidney tissues in DM group was much higher than those in NC group(P

Objective To study the effect of valsartan on expression of MCP-1 in the myocardium of diabetic rats,to investigate the protective effect of valsartan on the myocardium of diabetic rats.Methods 30 rats were divided into at random:normal control group(NC group),diabetic model group(DM group),diabetic model plus valsartan therapy group(DV group).Diabetic rats were induced by STZ,at 8th week,expression of MCP-1 in the myocardium of diabetic rats,and diabetic rats treated with valsartan was detected respectively by using immunohistochemistry.Results At 8th week,the expression of MCP-1 of DM group was much higher than in both DV group and NC group(P

Objective:Expression and purification of the α-HL of Staphylococcus aureus as antigen for making full human anti-α-HL antibody later ,providing of new treatment for Staphylococcus aureus infection.Methods:The total RNA of Staphylococcus aureus was extracted and the cDNA of α-HL was amplified by RT-PCR.The DNA of α-HL and pCold-TF plasmid was digested and ligated by T4 ligase and then transformed into E.coli TOPO 10.The recombinant plasmid α-HL/pCold-TF which verified by sequencing was trans-formed into E.coli BL21 for expression.The expression products was identified by SDS-PAGE and Western blot.Results: The size of amplified cDNA of α-HL was about 900 bp and the expressed soluble fusion protein of α-HL was about 90 kD(including the molecular chaperone in the vector ) after inducing expression for 24 h at 15℃.The Western blot results showed that the expressed protein was the fusion protein of α-HL.The purified α-HL was injected into BABL/c mice for making antiserum.The results showed that the antiserum had good binding activity with Staphylococcus aureus and the titer was greater than 10 000 times.Conclusion: The α-HL of Staphylococcus aureus was successfully cloned and the soluble fusion protein of α-HL was successfully expressed.

Objective:To single amino acid mutation of the full human scFvs against TSLP to enhance its affinity.Methods: The specific scFvs against TSLP was screened in our previous study and here the three-dimensional structures of TSLP and anti-TSLP scFvs were simulated by Discovery Studio system,then the molecular docking was made.The amino acids of binding epitope were randomly mutated and the mutated amino acids were selected which could remarkably improve the affinity of scFvs.The primers were designed based on the sequence of mutation amino acids and the scFv sequences were mutated by the overlapping extension PCR.The DNA of mutated scFvs was ligated with the expression vector pLZ16 and transformed into E.coli DH5αF′.Then the scFvs were expressed and the scFvs with improved affinity were selected by ELISA and BIAcore.Results: The five scFvs with single amino acid mutation were screened out by DS system,which could elevate the affinity of scFvs.The mutated anti-TSLP-scFvs were amplified by PCR,which size was about 1 000 bp.The mutated scFvs with correct sequence were expressed,and the mutated scFvs with improved affinity were detected by ELISA and BIAcore.The affinity of selected mutated scFv (M4) has been about 10 times higher than the scFv nonmutation.Conclusion: The affinity of anti-TSLP-scFv has been improved successfully.

Objective Construction of human IL‐4 recombinant expression vector and then conduct the expression ,purification and identification of human recombinant IL‐4 .Methods the open reading frame of IL‐4 was amplified by nest PCR with total RNA from PBMC of healthy volunteer .And then the amplified IL‐4 was inserted into pET101/D‐TOPO ,transformed into BL21 ,ex‐pressed ,purified and indentified .Results The size of amplified open reading frame of IL‐4 was about 460 bp and the sequence was correct .After transformed into BL21 ,the IL‐4 clone with higher expression level was selected by selection of different clones insert‐ed with IL‐4 and the size of expressed ,purified IL‐4 was about 28 × 103 .Western blot results showed that the size of single band was identical with the expected protein .Conclusion Human IL‐4 recombinant protein was got successfully .

Reflex tests are ordered when a particular test result indicates that additional testing should be performed according to the guidelines or the feedback process formulated by clinical consultation. The application scope of the reflex tests involves various subspecialties of laboratory medicine. The clinical application needs the support of qualified laboratory doctors, comprehensive information and financial system, clinical guidelines, and so on. Active application of reflex tests can promote the standardization of evidence-based medicine in clinical practice, save medical resources, and shorten the diagnosis and treatment time of patients.

Objective To explore the effect of transforming growth factor β1 (TGF-β1) on epigenetic histone lysine acetylation in the plasminogen activator inhibitor 1 (PAI-1) promoter and transcribe regions in glomerular mesangial cells (GMCs).Methods Chromatin immunoprecipitation assay and real-time quantitative PCR were used to detect Histone3K9 acetylation (H3K9Ac) in the PAI-1 promoter and transcribe regions induced by TGF-β1 and high glucose.Immunoprecipitation was also used to see the cooperation of Smad3,CBP and Sp1 proteins.Results In the four target regions of PAI-1 promoter,TGF-β1 treatment enhanced H3K9Ac at P1,P2 and P3 in GMCs (P ＜ 0.05),but no change was seen in the P4 region which was far from the transcription starting site.TGF-β1 obviously induced H3K9Ac in the T1 transcribe region of PAI-1 instead of T2 (P ＜ 0.05).High glucose increased PAI-1 mRNA expression and H3K9Ac around P1 promoter region (P＜ 0.05).TGF-β1 neutralizing antibody abrogated high glucose-induced H3K9Ac at PAI-1 promoter (P ＜ 0.01).TGF-β1 treatment could recruit Smad3 and CBP protein binding to the PAI-1 promoter regions (P1,P2,P3),and induce their cooperation in GMCs,which were responsing to TGF-β1 associated H3K9Ac.Conclusion TGF-β1 can induce H3K9Ac in the promoter and transcribe regions of PAI-1,promote Smad3 recruition and cooperation with Sp1 and CBP,which are associated with PAI-1 gene's regulation in GMCs.

Objective The aim of this study is to define if early change ofprocalcitonin (PCT) may inform about the efficacy evaluation of severe bacterial pneumonia.Methods A prospective,single-center,observational study was conducted in patients with severe bacterial pneumonia admitted to ICU in 2010 years.PCT samples were collected in baseline,72 hours,7 days and the ending in the duration of therapy.The efficacy evaluation was assessed at the end of treatment 5 days after and divided into the efficacy group and nonefficacy group according to the guiding principle of clinical research on antibacterial drugs by the Ministry of Health.Sixty-five patients with a mean age of (62.1 ± 15.9) years were evaluated.Five patients were severe community acquired pneumonia,32 patients nosocomial pneumonia and 28 patients ventilator associated pneumonia.The clinical pulmonary infection score(CPIS) was 7.9 ± 1.8 ;APACHE Ⅱ score was 14.5 ±5.3.There were 44 patients as the efficacy group and 21 patients as the nonefficacy group.SPSS13.0 was used to analyse the results.Results The PCT levels between efficacy group and nonefficacy group were (3.83 ±2.18)vs(4.23 ±2.64) μg,/L (t =1.249,P =0.387),(2.44 ± 1.05)vs(3.48 ± 1.75) μg/L(t=-1.959,P=0.045),(1.15 ±0.87) vs (3.41 ±1.58) μg/L (t=-2.904,P=0.006),and (0.51 ±0.17) vs (2.63 ±1.08) μg/L (t=-3.772,P =0.000) in baseline,72 hours,7 days and the ending in the duration of therapy.The change of PCT within the first 72 hours were (32.5 ± 12.4)％ vs (14.5 ± 7.1) ％.The area under receiver operating characteristics curve (AUC) of prediction clinical efficacy of the change of PCT within the first 72 hours was 0.823 (P =0.002),the AUC of white blood cell,the neuter granulocyte percentage,body temperature and PCT level within 72 hours were 0.575,0.543,0.521,0.597,respectively (P ＞ 0.05).In multivariate analyses,the change of PCT ＜ 30.8％ (odds ratio,15.2,95％ confidence interval,3.3-21.7,P =0.01) was independent risk factors of effect predictor.The changes of PCT within the first 72 hours (＞30.8％) combined with CPIS(＜6) were the best performance to predict clinical efficacy with a AUC of 0.910,sensitivity of 85.2％ and specificity of 92.5％.Conclusions The change of PCT within the first 72 hours can be used early to evaluate the effect in bacterial pneumonia.Especially,combined with CPIS can further improve the prediction value.