Objective:To clone soluble human anti-digoxin antibodies from large phage antibody library and construct a vector that expresses diabody.Methods:Soluble ScFvs were prepared through infecting E coli.HB2151 with the selected phage antibodies and induced with IPTG.The diabody vector was modified by enzyme digestion and checked by SDS-electrophoretogram.Results:It was showed by ELISA that soluble ScFv had specific binding ability to digoxin.The vector to express a soluble diabody was obtained by genetic modification,which was shown by Western blot.Conclusion:Soluble human anti-digoxin antibodies were successfully obtained from phage antibodies.The vector is efficient in creating diabody.

Objective: To establish an LC-MS/MS method for the determination of 4-( 4-amino-3-fluorophenoxy )-N-methylpyri-dine-2-carboxamide ( AFP-PMA) as a genotoxic impurity in regorafenib. Methods: The content of AFP-PMA was determined by an LC-MS/MS method. A Waters XBridge Shield RP18 column was adopted to separate the samples and the column temperature was 50℃. The mobile phase consisted of 5 mmol·L-1ammonium acetate aqueous (A)-acetonitrile (B) with gradient elution (0~9 min, 5%B→90%B) at a flow rate of 1. 0 ml·min-1. An electrospray ionization source (ESI) was used in a positive-ion and multiple reactions monitoring mode. The ion channel was m/z 262. 2→244. 1. Results:The standard curve was linear within the range of 2. 41-980. 90 ng·ml-1(r=0. 9998) and the limit of quantification was 8. 02 ng·ml-1. The limit of detection was 2. 41 ng·ml-1, which was e-quivalent to 0.000241% for the concentration of regorafenib. The average recovery was 100.95% and RSD was 2.37% (n=9). Conclusion:The method has good specificity, promising accuracy and high sensitivity, which can be used for determining the trace genotoxic impurity AFP-PMA in regorafenib.

Objective To explore clinical progress about the deep vein thrombosis after hip replacement at home and abroad,and discuss the preventive effect of nursing intervention.Methods The documents about the deep vein thrombosis after hip replacement were retrieved from PUBMED,China National Network Knowledge Infrastructure (CNKI),Chinese Scientific Journal Database (VIP),Chinese Bio Medical Literature Database (CBM) and Wan fang Database.And the retrieving duration was from 1st Jan.2001 to 1st Sept.2013.The research progress about prevention of deep vein thrombosis after hip replacement was analyzed through the analysis of the same and the difference of comparative literatures at home and abroad.Results There were totally 228 Chinese documents and 5 English documents of the deep vein thrombosis after hip replacement,and the number of document showed an increasing trend year by year.In the past five years,the clinical progress about the deep vein thrombosis after hip replacement was obvious,the research methods used was accurate,the involving intervention measures was continuously improved,literature quality was increased.Conclusions The quality of the documents published in domestic Chinese journals needs to be improved,and we should improve the quality of research design,research new methods of intervention,formulate the optimal nursing measures in order to achieve optimal care.

Objective To investigate the effects of volatile oil from artemisia dracunculus on myocardial injury caused by viral myocarditis in mice and explore its possible mechanism.Methods Totally 160 adult male BALB/c mice were randomly divided into normal control group (10) and viral myocarditis group (150).Viral myocarditis mice models were reproduced by intraperitoneal inoculation with a solution of coxsackievirus B3 (CVB3),a viral strain with affinity to myocardium,and then randomly divided into model,astragalus group,and low-,medium-,and high-dose volatile oil from artemisia dracunculus groups.After 1 hour of viral infection,normal control group and model group mice were given normal saline by intragastric administration,astragalus group mice were injected with astragalus 0.1 mL in each mouse by intraperitoneal injection,and the mice in other three groups were given low,medium and high dose (2％,5％,10％) 0.3 mL volatile oil from artemisia dracunculus in each mouse by intragastric administration,respectively,once a day for one week consecutively.The mortality,heart/body weight ratio,the activity of natural killer cells (NK cell),virus titer in myocardial homogenate,serum cardiac troponin Ⅰ (cTnI) level and myocardial pathological changes were observed.Results ① Mortality:the mortality of model group was higher than that of the normal control group,astragalus group,low and medium dose volatile oil from artemisia dracunculus groups (60.0％ vs.0％,23.3％,20.0％,28.7％),and the difference in the mortality being of no statistical significance between model group and that of high-dose volatile oil from artemisia dracunculus group (60.0％ vs.47.6％,P ＞ 0.05);the mortality of astragalus group was obviously lower than that of high-dose volatile oil from artemisia dracunculus group (P ＜ 0.01),and the differences in comparisons between the mortalities of astragalus intervention group,and medium-and low-dose volatile oil groups were not statistically significant (all P ＞ 0.05),and the comparison of mortality between low-and medium-dose volatile oil groups were also not statistically significant (P ＞ 0.05).② Immunization parameters:on the 8th day after modeling,the activity of NK cells in the model group was significantly lower than that in the normal control group [(15.91 ± 3.87)％ vs.(38.50 ± 2.32)％],the activities of NK cells in astragalus group,medium-and low-dose volatile oil from artemisia dracunculus groups were significantly higher than that in model group [(19.38 ± 3.27)％,(18.54 ± 3.09)％,(18.36 ± 2.64)％ vs.(15.91 ± 3.87)％,all P ＜ 0.05].None of virus was detected in the myocardial homogenate in the normal control group,and the virus titers in astragalus group,low and medium dose volatile oil from artemisia dracunculus groups were significantly lower than the titer of the model group (10-9/mL:1.96 ± 0.44,1.95 ± 0.46,1.95 ± 0.48 vs.2.41 ± 0.51,all P ＜0.01).③ Myocardial injury parameters:the level of cTnI in the normal control group was less than 0.1 μg/L,obviously lower than that in the model group [(15.84 ± 3.89) μg/L],as well as the ratio of heart/body weight in model group was also significantly higher than that in normal control group (× 10-4:8.3 ± 1.3 vs.4.6 ± 0.1),and the cTnI and the ratio of heart/body weight of astragalus intervention group,low and medium dose volatile oil from artemisia dracunculus groups were markedly lower than those of model group [cTnI (mg/L):10.03 ± 2.35,10.81 ± 2.56,11.10 ± 1.89 vs.15.84 ± 3.89,ratio of heart/body weight (× 10-4):7.2 ± 0.8,7.3 ± 1.0,7.3 ± 0.6 vs.8.3 ± 1.3].In the normal control group,there were no inflammatory cell infiltration and necrosis in myocardial tissue,the scores of myocardial pathological changes were 0.In the model group,the scores of inflammatory cell infiltration (3.25 ± 0.45) and of necrosis (2.91 ± 0.51) were markedly higher than those in the normal control group.And the above scores in astragalus group,low and medium dose volatile oil from artemisia dracunculus groups were significantly lower than those of the model group (infiltration score:2.92 ± 0.39,2.95 ± 0.35,2.95 ± 0.37 vs.3.25 ± 0.45,necrosis score:2.46 ± 0.50,2.50 ± 0.51,2.54 ± 0.50 vs.2.91 ± 0.51,all P ＜0.05).Conclusions Volatile oil from artemisia dracunculus can protect cardiomyocytes by removing the virus and regulating the immune function in the body.But the protective effects of volatile oil from artemisia dracunculus is related to the dosage,and the effects of low and medium dose are better.

A high performance liquid chromatography-tandem mass spectrometric ( HPLC-MS / MS) method coupled with an immunoaffinity clean-up column was successfully developed for determination of aflatoxins (AFB1 , AFB2 , AFG1 , AFG2 , AFM1 and AFM2 ) and zeranols ( α-zeranol, β-zeranol, α-zearalenol,β-zearalenol, zearalanone and zearalenone ). The sample was extracted with methanol-acetonitrile (20∶ 80, V/ V) after enzymatic digestion by β-glucuronidase / sulfatase, and the extraction solution was passed through glassy fiber filter paper and then diluted with phosphate buffer solution (PBS). The reconstituted solution was cleaned up with IAC-AZ immunoaffinity column, and then analyzed by HPLC-MS / MS in multiple reaction monitoring (MRM) mode. The results indicated that the linear detection range was 0. 03-6. 0 μg / L for AFB2 and AFG2 , and 0. 05-20 μg / L for the rest compounds. The correlation coefficients were above 0. 999. The limits of detection (LOD) and limits of quantitation (LOQ) were 0. 01-0. 03 μg / kg and 0. 04-0. 09 μg / kg, respectively. The recoveries of the aflatoxins and zeranols were in the range of 73. 6% -98. 4% at the spiked levels of 0. 5, 1 and 5 μg / kg, and the relative standard deviations (RSDs) were in the range of 1. 9% -11. 2% . The method was proved to be simple and accurate, and suitable for the rapid determination of aflatoxins and zeranols in animal-originated foods.

Objective To investigate the efficacy and safety of using various sedative regimens during transcatheter arterial chemoembolization (TACE) for hepatocellular carcinoma (HCC).Methods 156 HCC patients were randomly divided into four groups.The control group (n =30):intramuscular injection of 10 ml saline;The diazepam group (n =42):intramuscular injection of 10 mg of diazepam;The promethazine group (n =42):intramuscular injection of promethazine 25 mg;and The combined group (n =42):intramuscular injection of 10 mg of diazepam and promethazine 25 mg.Results The blood pressure and heart rate of the control group was significantly higher than the other three groups,while the combined group was significantly lower than the diazepam and the promethazine groups.The sedative rating:Grade 0 of the four groups were:30,11,18,0 patients respectively;Grade 1 of the four groups were:0,21,15,24 patients respectively;Grade 2 of the four groups were:0,10,9,18 patients respectively;No patients were in grades 3 and 4.The anxiety score of the control group was significantly higher than the other three groups.There was no significant difference in complications among the four groups.Conclusion Diazepam and/or promethazine could be used effectively and safely during TACE,and they could reduce stress response of HCC patients during TACE,and improved tolerance of TACE.

AIM: To investigate the role of Cx43 in the myocardialization of the proximal outflow tract(OFT) septum in the mouse heart.METHODS: C57/BL6 mice of ED11.5 to 1 day after birth were used in this study,which included Cx43 knockout homozygotes((Cx43-/-)),heterozygotes((Cx43+/-)) and wildtypes((Cx43+/+)).Pathohistological analysis was used to examine the structure of the hearts.The expression of alpha-sarcomeric actin(?-SCA),active caspase-3 and activator protein-2(AP-2) were detected by immunohistochemistry.RESULTS: Most(Cx43-/-) mice died within 24 h after birth with a swelling and blockage of the conotruncal region,which led to the obstruction of OFT and enlargement of right ventricle.HE staining showed plenty of abnormal tissues in this region forming many pouches.No apparent malformations were observed in(Cx43+/-) and(Cx43+/+) mice.The expression of ?-SCA in the proximal OFT septum was delayed obviously in(Cx43-/-).The apoptotic cells existed in the proximal OFT septum of(Cx43+/+) mostly during ED12.5 to ED15.5.However,there were less apoptotic cells observed in(Cx43+/-),and few in(Cx43-/-).The expression of AP-2,marker of neural crest cells,was increased in (Cx43-/-) and abnormally located in the proximal OFT septum.CONCLUSIONS: Cx43 KO mice are characterized by hyperplasia in conotruncal region,which may be associated with the delayed myocardialization of OFT septum.The decreased apoptosis and the abnormal distribution of cardiac neural crest cells are likely to contribute to the abnormal myocardialization in mice with Cx43 defects.

Objective To report experience of minimally invasive surgery of urinary calculi caused by melamine in infants. Methods Retrospectively reviewed the treatments and outcomes of 36 cases with urinary calculi caused by melamine from November 2007 to October 2008. 13 girls and 23 boys aged 8 to 36 months after daily consumption for six month or more of milk products tainted with melamine. These infants underwent MPCNL, ureteroscopic lithotripsy and placement of ureteral stent, respectively. Results The operations were performed successfully in all patients. Five cases underwent MPCNL. Ureteroscopic lithotripsy were performed in fourteen cases. Seventeen cases were placed of ureteral stents. No major complications like hemorrhea, perforation and organic injury were noted. The postoperative hospital stays were 3 to 10 days. All cases were followed up for 1 to 12 months. Calculus had no recurrence. Hydronephrosis and hydroureterosis disappeared or lightened. Growth and development were normal. Conclusions Various kinds of minimally invasive surgical procedures is safe and effective treatments for urinary calculi caused by melamine in infants, applicable in calculi with urinary obstruction especially.

Objective To investigate the efficacy and safety of endovascular treatment for subclavian steal syndrome (SSS). Methods During the period from April 2004 to Feb. 2013 at authors’ hospital, a total of 12 patients with SSS, including subclavian artery stenosis (n = 7) and occlusion (n = 5), underwent endovascular treatment. Percutaneous transluminal angioplasty (PTA) was carried out in 4 cases and stent implantation in 8 cases. The clinical data were retrospectively analyzed. Results Technical success rate was 100%(12/12). The diseased side/healthy side blood pressure ratio increased from preoperative (0.46 ± 0.28) to postoperative (0.89 ± 0.32), and the difference was statistically significant (P < 0.05). After the treatment, the clinical symptoms were improved in different degrees in all patients. During the follow - up period lasting for (16.7 ± 7.9) months, restenosis occurred in one case with left SSS thirteen months after PTA, and the condition was improved after stenting angioplasty. Another patient with severe subclavian artery stenosis developed peripheral micro- vascular thrombosis of the diseased limb in 23 days after PTA, which was relieved after medication to improve microcirculation and vasodilation. No severe complications such as stroke occurred. Conclusion For subclavian artery steal syndrome, endovascular management is a minimally invasive, safe and effective treatment. It can significantly relieve the clinical symptoms due to vertebral- basilar artery insufficiency caused by subclavian steal syndrome, and improve limb weakness, low body temperature, etc. Therefore, this technique should be recommended in clinical practice.