Child ; Clinical Laboratory Techniques ; Humans ; Tuberculosis ; diagnosis

AIM To express recombinant human cytochrome P450 3A4 mutants CYP3A4.3, CYP3A4.4, CYP3A4.5 and CYP3A4.18, and to employ them for in vitro metabolism studies of CYP3A4. METHODS Use Bac-to-Bac baculovirus expression system to recombinant baculovirus carrying cDNA of CYP3A4 mutants CYP3A4.3, CYP3A4.4, CYP3A4.5 and CYP3A4.18. Spodoptera frugiperda 9 (Sf9), cells were co-infected by recombinant viruses of CYP3A4 mutants, human NADPH-P450 oxidoreductase and cytochrome b5 to obtain recombinant proteins CYP3A4.3, CYP3A4.4, CYP3A4.5 and CYP3A4.18 with metabolic activity. RESULTS The mRNA transcription of CYP3A4 mutants in Sf9 cells were validated by RT-PCR. Testosterone and 7-benzyloxy-4-(trifluoromethyl) coumarin were metabolized by the lysates of Sf9 cells infected by the recombinant viruses. CONCLUSION CYP3A4 mutants CYP3A4.3, CYP3A4.4, CYP3A4.5 and CYP3A4.18 with metabolic activity were successfully expressed by baculovirus-insect cell expression system. The results indicated that recombinant CYP3A4. 5 showed lower activity comparing to the wild type protein towards testosterone, while CYP3A4. 18 with higher activity, and for CYP3A4.3 and CYP3A4.4 showing similar activity to the wild type protein.

Objective To evaluate the activity of antibiotics against pan-drug-resistant (PDR) Acinetobacter baumannii by combination antimicrobial susceptibility test in viro with epsilometric methods (Etest method) and microdilution checkerboard (CB method),and to detect a good correlation between timekill curve with the above mentioned two assays.Methods Thirty-one clinical isolates of PDR Acinetobacter baumannii were selected for mono and combination antimicrobial susceptibility test in vitro by E-test and CB method,then a comparison was conducted between the test results and the time-kill curve.Mono drugs involved tigecycline,colistin,imipenem and amikacin,and combinations involved two of drugs above,and three drugs involved imipenem/tigecycline,plus amikacin combination.Results Synergistic effect was detected in imipenem plus colistin and tigecycline plus imipenem combination.A high comparability was revealed between the E-test method with antimicrobial drugs added into the culture medium and the time-kill curves.Synergy in the combination of imipenem/tigecycline,plus amikacin was detected by the CB method and time-kill curves.Conclusion The results showed that the effect of specific combination of antibiotics against PDR Acinetobacter baumannii could be predicted by testing their synergistic effect with combination antimicrobial susceptibility test.

ObjectiveTo discuss the curative effects of sodium aescinate tablet on traumatic leg swelling.Methods168 cases were divided into four groups.54 cases of Ⅰ degree leg swelling were randomly divided into two groups,one group were treated with no drug and the other with sodium aescinate tablet.114 cases of Ⅱ and Ⅲ degree leg swelling were randomly divided into two groups,one group were treated with mannitol and the other with mannitol and sodium aescinate tablet.All cases were treated for 7 days and observed on leg swelling.ResultsThe curative effects of group with sodium aescinate tablet were superior to the group with no drug.The curative effects of group with both sodium aescinate tablet and mannitol are superior to group with mannitol.ConclusionSodium aescinate tablet had obviously curative effects on trautic leg swelling.Both sodium aescinate and mannitol could be used to treat leg swelling of Ⅱ and Ⅲ degree.

Objective To investigate the correlation of serum alpha-fetoprotein (AFP) with clinical pathological factors and prognosis of hepatocellular carcinoma ( HCC ).Methods Two hundred and twenty patients with HCC who hospitalized in our hospital from January 2007 to June 2009 were enrolled in this study.The relationship of AFP levels before operation to clinical pathological factors such as pathological grade,tumor size,tumor encapsulation,tumor numbers,portal vein tumor thrombus,TNM stage and postoperative recurrence were analyzed.Results Serum AFP concentration before operation were related with tumor size ( x2 =11.76,P ＜ 0.05 ),tumor encapsulation ( x2 =17.07,P ＜ 0.05 ),portal vein tumor thrombus ( x2 =8.944,P ＜ 0.05 ),higher TNM stage ( x2 =4.929,P ＜ 0.05 ) and early postoperative recurrence rate ( x2 =5.569,P ＜ 0.05 ).However,it had no correlation with pathological edmondson grade tumor numbers.Conclusion HCC patients with high serum AFP concentration often has greater tumor without encapsulation,high occurrence rate of portal vein tumor thrombus and higher TNM stage,they are more likely to early postoperative recurrence.Serum AFP before operation is useful not only for diagnosis,but also as a prognostic biomarker to predict prognosis and recurrence.

Objective To evaluate the activities of 18 pairs of antimicrobials combinations against non - duplicate clinical isolates of multidrug resistant Acinetobacter baumannii (MDRAB) in vitro.Methods Collect isolates of Acinetobacter baumannii from different patients from October 2009 to May 2010,which were isolated in Clinical Laboratory Center of Beijing Friendship Hospital,Capital Medical University.Use broth microdilution method to detect MIC of mono-antimicrobial,and checkerboard broth microdilution method to detect combinatied MIC,and calculate fractional inhibitory concentration (FIC) index to determine drug combinations effects.When the performance of the same drug combinations conflicted,appropriate strains were selected for screening of drug-resistant mechanisms by polymerase chain reaction( PCR),including efflux pump genes.Results In tests in vitro,rifampicin and polymyxin B,imipenem and gentamicin,cefepime and levofloxacin showed synergy at high proportion,68.1％,45.5％,40.9％,respectively.Minocycline and rifampicin,ampicillin/sulbactam and tobramycin.Ceftazidime and ciprofloxacin showed additive effect at high proportion,81.8％,68.2％,68.2％,respectively.There were several combinations which appeared the opposite effects to tested strains.Strains No.19 corresponding reaction was synergy and No.21,No.26 corresponding reactions were antagonism.The three strains above were selected for screening resistant mechanisms.The difference is that genotypes of adeS were negative in No.19 and positive in No.21 and No.26.Conclusion Rifampicin and polymyxin B combination showed synergy against the MDRAB in vitro,which can be considered as the treatment choice for critical infections caused by MDRAB.Imipenem and gentamicin,cefepime and levofloxacin also showed synergy in vitro,but in some isolates showed antagonism.This phenomenon may be due to the gene adeS activated by certain antibiotics,and the activated adeS drived efflux pump express or overexpress,which made the drugs in bacterial cells pumped out,causing antagonistic effect.The individual differences in strains should be considered when clinic strain apply these two combinations above.

Objective To evaluate the role of myosin light chain kinase(MLCK)in the increase in the permeability of human pulmonary microvascular endothelial cells(HPMVECs)induced by mechanical stretch.Methods HPMVECs were cultured in vitro and then randomly divided into 3 groups(n = 4 each): mechanical stretch group(group S),ML9 treatment group(group M)and control group(group C).In group S,the monolayer cells and magnetic beads coated with an RGD peptide were incubated in serum-free MCDB131 medium for 2 h,the unbound beads were washed out,and then the cells were exposed to magnetic twisting stimulation(MTS)for 2 h(frequency 3 Hz,intensity 4.2 mT).In group M,the monolayer cells and magnetic beads coated with an RGD peptide were incubated for 2 h in serum-free MCDB131 medium in which MLCK inhibitor ML9(50μmol/L)was added,and the other procedures were the same as in group S.In group C,the cells after washing as in group S were incubated for 2 h without exposing to MTS.Monolayer permeability was detected with FITC-dextran flux in transwell model.The distribution of integrin αVβ3 and actin was detected using the immunofluorescence assay.Results Compared with group C,the permeability of HPMVECs was significantly increased in group S,while no significant change was found in group M.Compared with group S,the permeability of HPMVECs was significantly decreased in group M.In addition,actin polymerized to form stress fiber,and integrin αVβ3 clustered at the end of stress fiber in group S,while actin mainly distributed in the surrounding area of the cell membrane and integrin αVβ3 evenly distributed on the cell surface in group C and M.Conclusion The mechanism by which mechanical stretch induces the increase in the permeability of HPMVECs is related to MLCK activation-mediated stress fiber formation and integrin αVβ3 clustering.

Objective To investigate the self-management self-efficacy,health-related quality of life of hemodialysis patients and explore the relationship between them. Methods Self-management self-efficacy questionnaire and SF-36 were used,and 55 hemodialysis patients participated in this study. Pearson correlate and regression analysis were used to test the relationship between them. Results 55 questionnaires were available. Positively associated between self-management self-efficacy and HRQOL(r=0.268,P＜0.05);significantly positively associated between physical activity self-efficacy and HRQOL(r=0.409,P＜0.01). Regression analysis showed that physical activity self-efficacy is the factor of HRQOL (t=3.261,P＜0.01).Conclusion Physical activity self-efficacy is the factor of HRQOL in hemodialysis patients,increasing self-management self-efficacy is benefit to their HRQOL.

It has long been concerned about the mechanism , pathology and toxicology of anaesthetic involving drug addiction and its influence on human body .In recent years , as a short non-coding RNA negatively regulating the genes in cells , MicroRNAs (miRNAs) may turn out to be a focus issue which is related to apoptosis , metabolism, inflammation, and tumorigenesis.This article illuminates the influence on gene expression introduced by common anaesthetics and the interaction between gene and its target site . Analyzing the potential medical applications of miRNA , this article reviews the correlation between miRNAs and anaesthetic to provide new evidences in this emerging field .

After 56 cases of infantile constipation were divided into the pattern of stagnation of heat in the stomach and intestines, pattern of obstruction of qi activity and pattern of deficiency in both qi and yin, the treatments were given by infantile Tuina techniques on the infantile points of Dachang (Extra),Zhongwan (CV 12), Bagua (Extra), Qijiegu (Extra), Chengshan (BL 57), Zusanli (ST 36) and Yingxiang (LI 20). The results showed cure in 45 cases, effectiveness in 9 cases, failure in 2 cases and the total effective rate was 96.5%.