Objective:To elucidate the effects of calcitonin gene-re la ted peptide(CGRP) on the gene expression of osteoprotegerin(OPG) and receptor ac tivator of nuclear factor-?B ligand(RANKL) in rabbit osteoblasts. Meth ods:Osteoblasts were cultured in media containing 10 -10～10 -7 mol/L of CGRP. After 24-hour incubation,semi-quanitative RT-PCR was perfor med to detect the expression of OPG and RANKL mRNA,and with ?-actin mRNA as th e internal control. Results:CGRP increased the mRNA expressio n of OPG with the maximal effect at the concentration of 10 -8～10 -7 mol/L. CGPR downgulated the mRNA expression of RANKL dose-dependantly.C onclusion:CGRP may regulate the activities of osteoclasts by regulating gene expression of OPG/RANKL.

Human gene therapy needs to express exogenous DNA at the targeting cells,producing a practical and efficient therapeutic dosage at an approp-riate time(quantitative pharmacology)with a safe man-ner.Recombinant adeno-associated virus(rAAV)Vec-tom possess a number of properties and recent progress in rAAV production made it rapidly become the reagent of choice for therapeutic gene tmasfer.Over 60 clinical trials of gene therapy based on rAAV have been carried out.The dose response reaction between rAAV vectors and gene expression activity or clinical outcome is one of major aspects of these trials.Most studies showed that vector genomes(vg)and gene expression had a concentration-dependent relationship during a certain scope.However,gene expr~sion Can be afffected by viral serotypes,tissue tropisms,cell targeting,drug regulation,injection route,age and sex,etc.Thus,these aspects should be carefully comidered by scienti-sts,pharmacologisis and physicians during animal ex-periments or clinical trails.KEY WORDS gene therapy;viral vector;dose-re-sponse;quantitative pharmacology;clinical thempy

Objective To analyze and evaluate the precision ,accuracy and linearity of the chemiluminescence method for detec‐ting plasma BNP .Methods According to the experimental schemes of CLSI EP15‐A2 ,EP6‐A files and other relevant documents , the precision ,accuracy and linearity of the Siemens Centaur XP chemiluminescence instrument for detecting BNP ,and the detection results were compared with the performance declared by manufacturer or the quality target formulated by laboratory .Results The imprecision of BNP detected by the chemiluminescence method was less than 1/3 TEa regulated by the Clinical Laboratory Center of Ministry of Health (allowable total error);the variation coefficient index (CVR) of internal quality control data was less than ± 2;the relative bias of the results of external quality control blind samples with the target values were less than Tea regulated by the Ministry of Health ;the linear evaluation results showed that BNP was once linearity in the range of 5 .3‐4696 .7 pg/mL .Conclusion The precision ,accuracy and linearity of the Siemens Centaur XP chemiluminescence instrument for detecting BNP can accord withthe quality objectives requirements and meet the clinical needs .

Objective To summarize the clinical features of the allergic bronchopulmonary aspergillosis (ABPA) in children,in order to improve the understanding for ABPA and make early diagnosis and treatment of the disease.Methods A retrospective study was performed on ABPA patients diagnosed in Department of Respiratory Medicine,Beijing Children's Hospital Affiliated to Capital Medical University from March 2010 to December 2013.The clinical features,laboratory results,image characteristics and the prognosis information were reviewed.Results Eight ABPA cases met the diagnostic criterion.All patients were school-age children (7 years and 2 months to 13 years and 8 months old).Cough (8 cases),productive sputum (8 cases),wheezing (5 cases),fever (4 cases) and hemoptysis (3 cases) were the main clinical features.Six of the 8 patients showed eosinophilia.IgE level was elevated in 7 patients (1.000-5.000 IU/L).All 8 patients were allergic to aspergillus fumigatus,while only 2 cases were positive in sputum culture for aspergillus fumigatus.CT scans showed pulmonary opacities in all 8 cases,while 7 patients had typical central bronchiectasis.Seven patients were treated with the regimen,which included glucocorticosteroid,antifungal agents (voriconazole or itraconazole) and regular bronchoscope.The symptoms of all treated patients relieved,the total serum IgE level and eosinophil cell count decreased spontaneously after the therapy.Conclusions ABPA is rare in children and the clinical features are non-specific.If the patient has elevated total IgE level in serum and eosinophilia,especially in patients with underling diseases,ABPA should be suspected.The positive result of specific antibodies to aspergillus fumigatus and central bronchiectasis on the radiology may give the suggestive diagnosis.ABPA patients generally have good response to the therapy of glucocorticosteroid and antifungal agents.

Objective:To construct K-ras-targeted siRNAs (K-ras siRNA) and to investigate the inhibitory effects of K-ras siRNAs on the growth and migration of lung cancer A549 cells (containing mutant K-ras gene) and NCI-H446 cells (containing wild-type K-ras gene). Methods: Four K-ras siRNAs (K-ras siRNA1～K-ras siRNA3 targeting wild-type K-ras and K-ras siRNA4 targeting mutant K-ras) were designed and artificially synthesized; they were used to transfect A549 cells and NCI-H446 cells. The expressions of Ras mRNA and protein were examined by RT-PCR and Western blot-ting. The inhibitory effects of K-ras siRNAs on the proliferations of A549 and NCI-H446 cells were determined by MTT assay. The effects of K-ras siRNAs on the cell migration and apoptosis were observed by Transwell assay and Hoechst 33258 staining, respectively. Results: Mutant K-ras-targeted siRNA (K-ras siRNA4) specifically inhibited the K-ras ex-pression but had no influence on H-ras and N-ras expression in A549 cells. K-ras siRNA4 inhibited the proliferation of A549 cells but did not inhibit that of NCI-H446 cells, which contained wild type K-ras gene. K-ras siRNA4 also induced apoptosis and inhibited migration of A549 cells. Conclusion: Mutant K-ras-targeted siRNA4 can inhibit the proliferation, migration and induce apoptosis of A549 cells. It may be a potential and personalized drug for the treatment against lung cancer containing mutant K-ras gene.

AIM: To investigate the effect of Shenmai Injection on the expression of bFGF and PCNA of gastric cancer in mouse. METHODS: RT-PCR was used to measure the expression of bFGF and PCNA in gastric cancer cell cultured with Shenmai Injection in three concentrations.immunohistochemistry used for protein synthesis in mouse gastric cancer model. RESULTS: 140 ?L/mL of Shenmai Injection concentration inhibited 63% of bFGF and PCNA gene expression. CONCLUSION: Shenmai Injection can inhibit the express of bFGF and PCNA.

Objective To study the cytogenetic features of acute myeloid leukemia (AML) and analyze the association with cytogenetic features and early responses after induction therapy.Methods The karyotypes of 395 patients who had been newly diagnosed with AML were analyzed.These patients were divided into three groups (low-risk,intermediate-risk and high-risk),according to the AML NCCN guidelines.The incidence of different karyotypes in these three groups and the complete remission (CR) rate after the first cycle of induction therapy were analyzed.Results The incidence rates of karyotypes in high-risk,intermediate-risk and low-risk groups were 50.56 ％ (180/356),39.89 ％ (142/356),9.55 ％ (34/356),respectively.All patients with t(15;17) who completed induction therapy reached CR.There was significant difference in the CR rates of t(8;21) groups with or without additional karyotypes [92.00 ％(23/25) vs 50.00 ％(11/22)] (x2 =10.317,P =0.001).There was no significant difference in the CR rates between normal and-Y karyotype group [61.90 ％ (39/63) vs 58.82 ％ (10/17)] (x2 =0.054,P =0.817).Complex cytogenetics ascribed to the low-risk group,of which monosomal karyotype was common,nine of ten patients with monosomal karyotype were associated with an inferior CR rate.Conclusion The cytogenetic features of AML are different from previous reports by other centers.The cytogenetic features of AML patients not only influence the long-term survival,but also the CR rates of induction therapy.

The mesaconitine and its major metabolites in the rat urine were identified by liquid chromatography and electrospray ionization tandem mass spectrometry. The rat urine was collected for consecutive 24 hours from the rat following intragastric infusion of mesaconitine, subsequently which were enriched and purified using solid phase extraction. The metabolites of mesaconitine in the rat urine were analyzed by the liquid chromatography and electrospray ionization tandem mass spectrometry. It is shown that the parent drug mesaconitine and its metabolites were found in the rat urine, such as hypo-mesaconitine glucuronic acid conjugate, 10-hydroxy-mesaconitine, 1-O-demethyl mesaconitine, deoxy-mesaconitine and hypo-mesaconitine. Among the five of metabolites, the hypo-mesaconitine glucuronic acid conjugate (m/z 766) was first discovered as the aconitine in rats phase II metabolites, which revealed a new way of mesaconitine metabolism in rats.

OBJECTIVE: To analyze the effects of Astragalous Injection on oxidative stress and micro-inflammatory status in patients undergoing maintenance hemodialysis (MHD). METHODS: Sixty MHD patients were included and randomized into treatment group and control group, with another 10 healthy volunteers as normal control. The patients in the treatment group were treated with Astragalous Injection and the patients in the control group were treated with normal saline for 12 weeks. A spectrophotometric method was used for the measurement of plasma concentrations of oxidative parameters including advanced glycation end products (AGEs), advanced oxidation protein product (AOPP), malondialdehyde (MDA) and vitamin E (Vit E). The content of C-reactive protein (CRP) was evaluated by enzyme-linked immunosorbent assay. RESULTS: Compared with the normal control group, the plasma levels of AGEs, AOPP, MDA and CRP were significantly increased, while plasma level of Vit E was significantly decreased in MHD patients ( P<0.01). After Astragalous Injection treatment, the plasma levels of AGEs, AOPP, MDA and CRP were decreased as compared with the control group ( P<0.01), while there was no significant difference in plasma Vit E level between the treatment group and control group. CONCLUSION: There exist oxidative stress and micro-inflammation in MHD patients. Astragalous Injection can ameliorate the accumulation of oxidative products and micro-inflammatory status, but it has no significant effect on plasma Vit E level.

How to discover the valuable knowledge from the large amount of prescription data accumulated in the long-term medical practice of traditional Chinese medicine (TCM) is one of the important contents of TCM modernization.Based on the prescription data of Xue' s clinical practice of many years,this paper explored the method of combining TCM network and prescription network to find out Xue' s common core drug combination.Based on 9,584 prescriptions in the Hospital Information System of Guang'anmen Hospital of China Academy of Chinese Medical Sciences,prescription network and drug network were constructed according to the similarity of prescription composition and drug co-occurrence relationship.Using the complex network analysis methods,such as community analysis method,to analyze the prescription and drug compatibility,the results were evaluated and analyzed by Xue and his successors.As a result,through complex network analysis,126 modular prescriptions and 4 TCM modules were obtained.One of the core components of the prescription module included Xiao Chai Hu decoction,Yin Qiao powder,and Sheng Jiang powder compound addition and subtraction.It was in consistent with the drug composition of exogenous febrile prescriptions excavated earlier.In conclusion,using the complex network methods,we can get some core drug combinations prescribed by Prof.Xue,and achieve the common compound core drug combination for treating diseases with certain vantages,laying a foundation for further inheriting and excavating Xue' s effective experience.