OBJECTIVE:To introduce the experience of application and report for accreditation of drug clinical trial institution qualification of our hospital,to provide a reference for other hospitals. METHODS:From the building of institution,drug clinical trial ethics committee,clinical trials of listed drugs,simulated qualification accreditation,and other aspects,the experience and key points of application and report for accreditation of drug clinical trial qualification of our hospital were summed up. RESULTS& CONCLUSIONS:Priority should be given to staffing(particularly the leaders of the institution),the installation of software and hardware (for system development,training,institution office and specialized department),and the building of ethics committee. Departments should share work experience of clinical trials of listed drugs and set up a self-inspection group to simulate qualifica-tion accreditation. Work procedures and methods of qualification accreditation should be clearly understood,the requirements of Good Clinical Practice,the standards for qualification accreditation,should be known well;Good Clinical Practice should be seri-ously implemented and the subjects’rights and interests should be protected,which will contribute to the successful completion of application and report for qualification accreditation.

Objective To study the effect of the serum levels of 25-hydroxyvitamin D3 [25-(OH)D3] and total IgE (TIgE) in pathogenesis of the children bronchial asthma.Methods Twenty-two children with bronchial asthma (bronchial asthma group) and 20 healthy children (control group) were selected.The serum levels of 25-(OH)D3 and TIgE in the children of 2 groups were detected.Results The level of serum 25-(OH)D3 in bronchial asthma group was significantly lower than that in control group [(23.64 ± 3.89)μ g/L vs.(35.82 ± 4.37) μ g/L],there was statistical difference (P ＜ 0.05).The level of serum TIgE in bronchial asthma group was significantly higher than that in control group [(208.62 ± 32.59) kU/L vs.(73.84 ± 18.86) kU/L],there was statistical difference (P ＜ 0.05).There was negative correlation between the level of serum 25-(OH)D3 and TIgE in bronchial asthma group (r =-0.832,P ＜ 0.01),but there was no correlation between the level of serum 25-(OH)D3 and TIgE in control group (r =-0.038,P ＞0.05).Conclusions The lack of 25-(OH)D3 may be associated with allergy and childhood bronchial asthma.By monitor the levels of 25-(OH)D3 and TIgE may assess condition of childhood bronchial asthma.

Objective To investigate the expression patterns of artemisinin biosynthetic genes in Artemisia annua L.during the development stage and in different tissues,and to explore the mechanism of spatial and temporal modulators for artmisinin production.Methods The transcriptional profiles of artemisinin biosynthetic genes in the capital and cooperative pathways were quantitatively assayed by real time fluorescence quantitative-polymerase chain reaction(RTFQ-PCR) in different tissues of roots,stems,leaves and flowers,and in leaf-flourishing,pre-floral,and post-floral periods.Results The expression levels of the tested genes were extremely low in June,raised in July,and reached their peak values in August(before flowering),but dropped gradually in September(after blooming).In August,the transcription levels of the tested genes increased by 3 to 15 times compared to the lowestlevels.In particular,ADS and CYP71AV1 mRNA levels had the great elevation,which were 12 and 15 times as much as those in June.During the flowering period,the artemisinin biosynthetic genes mRNA expression was detectable in roots,stems,leaves and flowers,and the expression levels had no obvious difference except that ADS mRNA level in leaves was 2 times higher than that in other tissues(P

ObjectiveTo observe the effect of the combined therapy of ginkgo-dipyridimolum and mecobalamin for diabetic peripheral neuropathy (DPN).MethodsA total of 102 DPN cases were randomly divided into treatment group(n =50) and control group(n =52).The treatment group was given ginkgo-dipyridimolum and mecobalamin, and the control group was given only mecobalamin for 4 weeks.We observed the change of symptom and sign of both groups before and after treatment,and the MNCV and SNCV of median nerve,ulnar nerve and peroneal nerve were detected.ResultsIn the treatment group, the improvement of DPN symptoms (x2 = 10.090, P ＜ 0.01) and nervous conducting speed were much more obvious than that in the control group(MNCV of median nerve, ulnar nerve and peroneal nerve was(49.12 ± 3.32) m/s, (52.11 ± 3.12) m/s and(47.32 ± 3.21) m/s respectively; SNCV of median nerve,ulnar nerve and peroneal nerve was(47.13 ± 2.56) m/s, (55.12 ± 2.31) m/s, (46.78 ± 2.66) m/s respectively,all P ＜0.05).ConclusionThe combined therapy of ginkgo-dipyridimolum and mecobalamin could effectively improve the therapeutic outcome of DPN, and the treatment was safe and worthy of widespread use.

The effect of nateglinide or sequential treatment with metformin on glycemic stability in newly diagnosed type 2 diabetes was investigated. Thirty-four cases of newly diagnosed type 2 diabetes received nateglinide therapy, or sequential treatment with metformin according to fasting and postprandial blood glucose, and were classified into isolated nateglinide therapy group(n=14) and sequentially treated with metformin group(n=20). Glycemic stability, reflected by mean amplitude of glycemic excursions(MAGE) and HbA1C, was determined in all patients before and after therapy for three months. HbA1C and MAGE in two groups were all improved after treatment(P＜0.05). The therapy of nateglinide alone or combined with metformin can significantly improve glycemic stability in newly diagnosed type 2 diabetes.

Objective To investigate the value of measurement of cardiac output in children by bio-reactance versus echocardiography.Methods Pediatric patients admitted in pediatric department of Peking University First Hospital from September to December 2012 who needed hemodynamic monitoring were enrolled prospectively.Cardiac index(CI)and stroke volume(SV)were measured by echocardiography and non-invasive cardiac output measurement(NICOM)and compared by Spearman correlation and Bland-Alt-man analysis.Results Thirty patients were included.The median age was 7.25 years.CI[M(P5 ,P95 )] measured by NICOM and echocardiography were correlated significantly[3.42(2.28,4.92)L /(min?m2 ) vs.3.51 (2.94,4.85 )L/(min?m2 ),R =0.385,P =0.035 ].Bland-Altman analysis revealed a bias of-0.22 L/(min?m2 )(P =0.051 ),limits of agreement of -1.40 to 0.95 L/(min?m2 ).SV[M(P5 ,P95 )] measured by NICOM and echocardiography were correlated more significantly [36.3 (12.6,87.8 )ml vs.39.4(14.7,86.9)ml,R =0.768,P ﹤0.001 ].Bland-Altman analysis revealed a bias of -3.1 ml(P =0.176),limits of agreement of -27.4 to 21.2 ml.Conclusion There is no significant difference between NICOM and echocardiography for the measurement of CI and SV in pediatric patients.Further validation studies need to be conducted before routine clinical use.

Objective To increase artemisinin yield in transgenic Artemisia annua plants by regulating metabolic affluxion through metabolic pathway engineering. Methods The gene targeting vector was constructed by squalene synthase (SS) gene of A. annua, green fluorescent protein (GFP) gene, and cytosine deaminase (CodA) gene, and the vector was introduced into Agrobacterium tumefaciens by freeze-thawing procedure. A. annua was transformed through Leaf Disk method and regenerated transgenic plants were screened by the “Step-by-Step Selection”. Results Among the transgenic A. annua plants emitting green fluorescence after expression of GFP gene, the exogenous GFP gene rather than endogenous SS gene was detected in one transgenic plant by PCR as well as hybridization of PCR products. The preliminary data showed that the wild-type SS gene was replaced by mutated SS gene in the transgenic A. annua plant. Conclusion Gene targeting of squalene synthases of A. annua is successful.

OBJECTIVE:To develop a method for the concentration determination of olanzapine,risperidone and paliperidone in human plasma.METHODS:After liquid-liquid extraction,using buspirone hydrochloride as internal standard,the concentration of plasma sample was determined by UPLC-MS/MS.The determination was performed on ACQUITY UPLCTM BEH C18 column with mobile phase consisted of methanol-0.01 mol/L ammonium formate solution (gradient elution) at flow rate of 0.2 mL/min.The column temperature was 45 ℃,and sample size was 5 μL.The electrospray ionization source was adopted for positive ion scanning under MRM mode.Ion-pairs for quantitative analysis were as follows:m/z 313.29→256.25 (olanzapine),m/z 411.42→191.19 (ris peridone),m/z 427.45→207.18 (paliperidone) and m/z 386.43→122.37 (internal standard).RESULTS:The linear ranges of olanzapine,risperidone and paliperidone were 0.426-108.954,0.213-54.476,0.213-54.476 ng/mL,respectively.RSDs of inter-day and intra-day were all lower than 20％.The recoveries of them ranged 83.3％-112.9％,90.0％-109.8％ and 95.2％-114.9％,respective ly.Extraction recoveries ranged 65.5％-95.0％,73.9％-98.5％ and 73.6％-99.4％,respectively.Both plasma matrix effect and dilute effect didn't influence the determination of plasma concentration.The plasma concentrations of olanzapine,risperidone and paliperidone in 100 schizophrenia patients were (103.3 ± 73.6),(13.1 ± 13.1) and (23.2 ± 20.0) ng/mL,respectively.CONCLU SIONS:The method is simple,rapid,sensitive and specific.It can be used for the determination of plasma concentration and pharmacodynamic study of olanzapine,risperidone and paliperidone.

Objective To explore the feasibility of utilizing the cytosine deaminase A (CodA) gene as an effective negative selectable marker in Artemisia annua for gene targeting.Methods The PCR procedure was employed to amplify CodA gene from Escherichia coli.After being cloned and sequenced, the gene was inserted into a plant expression vector, pROKⅡ, and then introduced into Agrobacterium tumefaciens LBA4404 (pAL4404).The leaf disks of A.annua were transformed by the co-cultivation protocol, after which the transformed calli were selected and green shoots of A. annua were regenerated on N6 medium supplemented with 25 ?g/mL Kanamycin (Kan).Then the Kan-resistant transgenic shoots were transplanted onto the MS medium containing 500 ?g/mL 5-fluocytosine (5-FC) plus 25 ?g/mL Kan and continuously cultured for up to two weeks.Results The transgenic shoots have totally died while untransformed shoots still kept normal growth, indicating that A.annua cells introduced into the CodA gene had conferred an expected negative selection phenotype.When detected by RT-PCR, the transgenic shoots displayed a CodA-positive amplified band, but untransformed shoots gave no such CodA-specific amplified pattern.This result suggested that CodA gene had transcribed into corresponding mRNA in A.annua cells with furtherly verifying the result of phenotypic assay.Conclusion The CodA gene can be utilized as an effective negative selectable marker in A.annua for gene targeting.