In the present study, the expression, purification, toxicity detection and the adjuvant effect of the heat-labile enterotoxin in non-toxic mutant mLT63 of Escherichia coli were investigated, in which the inductive expression was performed under optimal condition for inductive expression and the toxicity of the products obtained from inductive expression were tested for toxicity after being purified and concentrated with affinity chromatography. BALB/c mice were immunized orally with the mutant mLT63 associated with Helicobacter pylori (Hp) subunit vaccine UreB, Omp11. After immunization, the specific antibody levels in serum, extract from gastric tissues and fecal extract were determind by means of ELISA assay and the results were subjected to statistical analysis. It was demonstrated that the mutant mLT63 of heat-labile entrotoxin of E.coli constructed in our laboratory devoided of any toxic effect as revealed by the rabbit ileal loop assay, but its adjuvant effect could be demonstrated in the associated immunization of mice with Hp subunit vaccine UreB and Omp11.

Objective:To study the stability of compound Kushen mixed liquid to provide reliable evidence for safe clinical drug application.Methods:Compound Kushen injection was mixed respectively with 0.9% sodium chloride injection and 5% glucose injection,and then placed under the different conditions for 0,1,2,4,8,12,24 and 48 h.HPLC was conducted to determine the content changes of four components in compound Kushen mixed liquid,and the appearance,pH and insoluble particles were observed as well.Results:The mixed liquid of compound Kushen with 0.9% sodium chloride injection was stable in 48 h without the influence of light and temperature.However,the mixed liquid of compound Kushen with 5% glucose injection had poorer stability with storage time shorter than 12 h at room temperature and 2 h at high temperature.Conclusion:The stability of the mixed liquid of compound Kushen is closely related to the pH value of solvent.0.9% Sodium chloride injection is recommended as the solvent,and the mixed liquid should be used up in 48 h.

Objective To evaluate the correlation and difference of reversed phase high performance liquid chromatography (RP-HPLC) and fluorescence polarization immunoassay (FPIA) on determining serum concentration of carbamazepine.Methods Fifty serum samples were collected,both RP-HPLC and FPIA methods were employed to determine the concentration of carbamazepine.The results were analyzed by paired t test,Bland-Altman and Deming regression methods,respectively.Results The results of measuring 50 samples by the two methods showed that FPIA datas were significantly higher than RP-HPLC datas,and there was statistically significant difference(P<0.05) and poorer consistency between two methods;There was good correlation between carbamazepine concentrations determined by the two methods.Deming regression equation was CFPIA=1.195 3 CRP-HPLC-0.144 0,and Pearson correlation coefficient was 0.968 5.Conclusion Clinicians should pay more attention to the difference of carbamazepine concentration determination by different methods when carbamazepine individualized dosage regimen was adjusted according to therapeutic drug monitoring.

OBJECTIVE:To investigate the compatible stability of Calcium folinate for injection mixed with Glucose injection and Sodium chloride injection. METHODS:Referring to clinical common concentration,each 3 Calcium folinate for injection (each injection was equal to calcium folinate 100 mg)were respectively mixed with Glucose injection 250 mL or Sodium chloride injection 250 mL. At room temperature,under light or dark condition,the appearance of mixtures,pH value and the number of in-soluble particles were investigated 0,1,2,3,4,6,8,12,24,36,48 h. The contents of calcium folinate in mixtures were deter-mined by HPLC. RESULTS:Under above condition,the color of the mixtures had no change,and no gas,precipitation and turbid-ity was found;there was no evident change in pH values(RSD<2%,n=11). 0 h after mixing,there was large number of parti-cles≥10 μm in mixtures,but the number of particle was decreased as time;within 48 h,the number of particles ≥10 μm and ≥25 μm in mixtures were all in line with the standard of Chinese Pharmacopeia(2015 edition). Under the protection from light con-dition,relative contents of calcium folinate in mixtures had no significant change(RSD<2%,n=11). Under light condition,rela-tive contents of calcium folinate in mixtures decreased significantly,decreasing to 94.5%(mixed with Glucose injection) and 88.4%(mixed with Sodium chloride injection). CONCLUSIONS:Calcium folinate for injection is more stable in Glucose injec-tion,and the stability of compatibility can be affected by light conditions. After mixed with Glucose injection and Sodium chloride injection,Calcium folinate for injection should be kept away from light and used as soon as possible

Objective To develop a rapid ultra high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/ MS) for determination of 11 sedative-hypnotics in human plasma. Methods The plasma samples were extracted with dichloromethane:n-hexane:acetoacetate (5∶4∶1). The separation was performed on a Waters ACQUITY UPLC HSS T3 column (2.1 mm×100 mm,1.8 μm) using the mobile phase composed of acetonitrile-0.1% ammonium solution at a flow rate of 0.2 mL?min-1 in gradient elution mode. The detection was performed on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring (MRM) via electrospray ionization (ESI) source in positive mode. Results Good linear relation was obtained over the investigated concentration range, with all correlation coefficients higher than 0.99. The limit of detection was 200 pg?mL-1 for dexzopiclone,and 10- 20 pg?mL-1 for other sedative-hypnotics. The intra- and inter-day RSD of 11 sedative-hypnotics were no more than 15.0%. The recoveries were in the range of 72.3%-108.0%, and the matrix effects were approximately 0.86- 1. 12. Conclusion The method is rapid, sensitive and reliable, and suitable for the simultaneous determination of 11 sedative-hypnotics in human plasma.

OBJECTIVE:To investigate the compatible stability of Xiao'aiping injection combined with 3 kinds of common in-jections. METHODS:Referring to package inserts,Xiao'aiping injection 40 mL was compatible with 5% Glucose injection,10%Glucose injection or 0.9% Sodium chloride injection 160 mL,respectively. At room temperature(about 25 ℃)and high tempera-ture(40 ℃),the appearance of mixtures were observed at 0,1,2,4,8,12,24,48 h;pH value and the number of insoluble particles were detected. The contents of tenacissoside A and tenacissoside Ⅰ in mixtures were determined by HPLC. RESULTS:Un-der above condition,the mixtures were brownish yellow liquid within 48 h after Xiao'aiping injection was compatible with 5%Glucose injection or 10% Glucose injection;24 h after mixed with 0.9% Sodium chloride injection,the mixture changed from brownish yellow to reddish brown,but no precipitation was found. The pH value of mixtures had no significant change(RSD<1%,n=8). The number of particles ≥25 μm was in line with the requirements of Chinese Pharmacopeia(2015 edition). For-ty-eight hours after mixing,the number of particles ≥10 μm in the mixtures exceeded the pharmacopoeia limits. Within 48 h after mixing,the relative contents of tenacissoside A and tenacissoside I in mixtures had no significant change(RSD<2%,n=8). CON-CLUSIONS:The mixture should be used up within 24 h after Xiao'aiping injection combined with 5% Glucose injection,10%Glucose injection or 0.9% Sodium chloride injection.

OBJECTIVE:To develop a method for rapid,accurate and simultaneous determination of dexzopiclone,zolpidam and zaleplon in human plasma. METHODS:After the plasma sample was processed by liquid-liquid extraction,UPLC-MS/MS was established to detect plasma sample with carbamazepine as internal standard. The separation was performed on a Waters ACQUITY UPLC HSS T3 column with mobile phase composed of 0.1% ammonium solution-acetonitrile (gradient elution) at flow rate of 0.2 ml/min. The column temperature was set at 40 ℃,and sample size was 5 μl. The detection was performed by multiple reaction monitoring (MRM) via electrospray ionization (ESI) source in positive mode. The mass transition ion-pairs were as follows:m/z 308.2→263.1(zolpidem),m/z 389.3→245.0(dexzopiclone),m/z 306.2→236.1(zaleplon),m/z 237.3→151.2(internal standard). RE-SULTS:The linear range of zolpidem,dexzopiclone and zaleplon were 0.02-20.00,0.50-20.00,0.02-20.00 ng/ml,respectively (r=0.990 1,0.996 8,0.991 7). LLOQ of them were 0.02,0.50,0.02 ng/ml,and detection limit were 0.01,0.20,0.01 ng/ml. RS-Ds of intra-day and inter-day were all lower than 15% ;extraction recovery were 88.9% -106.5% ;matrix effect were 94.8%-106.3%. CONCLUSIONS:The method is simple,rapid,sensitive and specific,and it can be used for simultaneous deter-mination of zaleplon,zolpidem and dexzopiclone in human plasma.

Physical restraint is a complex and dynamic process in which nurses are the main decision makers and implementer. This article is to review the factors influencing the implementation of physical restraint in ICU nurses, and summarize from 4 aspects: nurses' own factors, environmental factors, promotion measures of rational use of physical restraint and ethical factors, so as to provide a basis for the effective clinical intervention measures in the future.

OBJECTIVE:To investigate drug combination in outpatient prescriptions of hypertension patients in our hospital, provide reference for rational drug use in clinic. METHODS:The outpatient prescriptions of patients diagnosed as hypertension dur-ing Jan. 1st to Feb. 1st in 2015 were collected from the hospital. The prescriptions of two or more than two drugs were screened, and the prescriptions of drug combination containing CYP enzyme substrate,inhibitor or inducer were recorded. Guided by metabol-ic enzymology theory,the potential metabolic drug interactions in prescriptions were evaluated on the basis of relevant literature and data reports. RESULTS:Totally 1042 prescriptions were consulted. The prescriptions of the combined medication were 551, and the potential metabolic drug-drug interactions were detected at 249 prescriptions,accounting for 45.2％. Main CYP enzyme sub-types were CYP3A4,CYP2C9,CYP2C19 and CYP2D6. Totally 214 prescriptions were correlated with CYP3A4,accounting for 85.9％ of drug interaction prescriptions;CYP3A4 substrate combined with substrate in 199 prescriptions,with inhibitor in 27 pre-scriptions,and with inducer in 11 prescriptions. Totally 27 prescriptions were correlated with CYP2C9,accounting for 10.8％ of drug interaction prescriptions;CYP2C9 substrate combined with substrate in 8 prescriptions,and with inhibitor in 20 prescriptions. Totally 27 prescriptions were correlated with CYP2D6,accounting for 10.8％ of drug interaction prescriptions;CYP2D6 substrate combined with substrate in 15 prescriptions,and with inhibitor in 12 prescriptions. Totally 4 prescriptions were correlated with CYP2C19,accounting for 1.6％ of drug interaction prescriptions;CYP2C9 substrate combined with inhibitor in 2 prescriptions, and with inducer in 2 prescriptions. CONCLUSIONS:Many metabolic drug-drug interactions are detected in the outpatient prescrip-tions of hypertension patients in our hospital. In order to improve the rationality and safety of the prescription,clinicians and phar-macists should pay attention to the drug combinations with drug-drug interactions which have been reported in the existing litera-ture,and choose similar drugs without or with little interactions.

2019 novel coronavirus(2019-nCoV) outbreak is one of the public health emergency of international concern.Since the 2019-nCoV outbreak, China has been adopting strict prevention and control measures, and has achieved remarkable results in the initial stage of prevention and control.However, some imported cases and sporadic regional cases have been found, and even short-term regional epidemics have occurred, indicating that the preventing and control against the epidemic remains grim.With the change of the incidence proportion and the number of cases in children under 18 years old, some new special symptoms and complications have appeared in children patients.In addition, with the occurrence of virus mutation, it has not only attracted attention from all parties, but also proposed a new topic for the prevention and treatment of 2019-nCoV infection in children of China.Based on the second edition, the present consensus further summarizes the clinical characteristics and experience of children′s cases, and puts forward recommendations on the diagnostic criteria, laboratory examination, treatment, prevention and control of children′s cases for providing reference for further guidance of treatment of 2019-nCoV infection in children.