Objective To explore effect of CYP3A4 gene polymorphism and genetic testing on the efficacy of fentanyl in patients with extensive burns.Methods A prospective randomized controlled study, 149 patients with extensive burns were picked from Feburary to July in 2015.Patients were randomly divided into matched group ( n=56 ) and experimental group ( n=93 ) . The matched group was treated with fentanyl 5 g/kg to finish implementation of anesthesia induction.Depends on the gene detection results, the patients in experimental group were treated with 6 g/kg, 5 g/kg or 4 g/kg fentanyl.The VAS score at different time after operation in the experimental group and the control group were compared , and the dosage of fentanyl was observed.Patients whose VAS score was greater than 7 should treated with intramuscular injection of pethidine 100 mg, recorded all patients with pethidine,s additional cases and the times.At the same time, the adverse reactions of the two groups were recorded before and after hospital discharge. ResuIts The score of VAS was not significantly different after wake up immediatly after the surgery , and the score of VAS was significantly lower in the experimental group patients in 6h,12h after the surgery (P<0.05).The dosage of fentanyl in the experimental group and the number of the cases and the times were significantly decreased (P<0.05).The adverse reactions after the surgery such as nausea, vomiting, itching, drowsiness, and adverse reactions were significantly lower in the experimental group (P<0.05).ConcIusion CYP3A4 gene detection in patients with large area of burn in the individual drug use is important, because of patients’ genotype adjusting fentanyl dosagecan, it enhances significantly the analgesic effect, reduces the amount of drug use, and effectively reduces the adverse reactions.

Objective To develop an effective process for isolating and purifying haptoglobin ( Hp) from Cohn fractionⅣby a new ion exchange chromatography and to preliminarily identify and analyze the product of each purification step . Methods The fraction was first diluted and impurities were adsorbed with Rivanol .Then, the supernatant was treated with 50%ammonium sulfate.Finally, the precipitate was redissolved , and Hp was purified further with Q Sepharose Fast Flow chromatography .Native-PAGE was used to measure the activity of the haptoglobin-bound hemoglobin , while SDS-PAGE analysis and immunoblot were used for identification of the target protein .Results After pretreatment , some of the impuri-ties were removed from the Cohn fraction Ⅳ, and the target protein was enriched .In our case, the target protein was Hp and Hp2-2 was the main phenotype in the human plasma fraction Ⅳ.Target protein band and high purity were identified by SDS-PAGE.Immunoblot analysis further proved that this method could successfully isolate the target protein Hp , and the activity of 2.8 U/ml was measured by Native-PAGE method.Conclusion Haptoglobin is successfully isolated from human Cohn fractionⅣwith this method.The purification process is simple and suitable for scale-up production with a good prospect.

Plasma products are considered to be special medicinesderived from healthy human plasma .During 1980′s, events of transmission of human immunodeficiency virus through plasma products were frequently reported .Since then, ensuring the viral safety of plasma products has raised great concerns all over the world .So far, with decades of effort , most countries in the world have established rigorous systems with preventive measures to ensure the viral safety of plasma prod -ucts.These measures include control of source plasma , validated inactivation/removal of infectious agents , the adherence to current good manufacturing practices .Nevertheless , new infectious agents which may be threats to viral safety require continuous studies on appropriate countermeasures .

Objective To establish an assay for detecting α2-macroglobulin activity in Cohn fraction Ⅳ.Methodsα2-M reacted with trypsin to form α2-M-trypsin complex.After the chromogenic substrate Na-benzoyl-DL-arginine 4-nitroanilide hydrochloride ( BAPNA) was added, absorption at 410 nm was detected with the microplate reader .α2-M activity in Cohn fractionⅣwas quantitatively detected according to the established standard curve of plasma α2-M activity. Result Several critical parameters in this assay were optimized .A standard curve of plasma α2-M activity was established . According to this standard curve ,α2-M activity in Cohn fraction Ⅳsample was detected to be 1.578 PU/ml.Conclusion Using normal human plasma as the reference material , theα2-M activity in Cohn fractionⅣcan be detected through chro-mogenic substrate assay.This study provides a simple method to detect α2-M activity during the purification process of α2-M from Cohn fraction Ⅳ.

Objective To promote the progress in varicella-zoster virus (VZV) immunoglobulin preparation,a rapid microneutralization test ( RMNt) was set up for screening plasmapheresis donations with high titers of special neutralizing antibodies to VZV.Methods With reference to the VZV immunoglobulin (VZIG) preparation standard of FDA and VZIG international unit ( IU) , a screening standard was formulated; the amount of virus was analyzed to determine the optimal conditions for RMNt;screening technology was established and the IU was introduced as quality control ;twenty samples of apheresis plasma and fifteen samples of pooled plasma were diluted at 1∶2 to 1∶256 and tested by RMNt respectively;and the sensitivity of RMNt was also analyzed by the commercial ELISA kit .Results Plasma samples that were diluted at 1∶16 and had a titer more than 0.4 U/ml could be used in the production of VZIG .1500 PFU/ml titers of virus in RMNt pro-duced readable results in plasma screening .Eight apheresis plasma samples tested met the screening standard , but none of the pooled samples tested positive .RMNt had a good linear relationship with ELISA (r=0.895 24,P<0.0001).Conclu-sion The sensitivity, throughput and operability of the established RMNt can be used in the screening of plasma donations as key techniques for the production of VZIG .

Objective To establish and evaluate a universal real-time fluorescent quantitative PCR(qPCR)method for identifying and quantifying three human parvovirus B 19 ( B19V) genotypes.Methods Firstly, following a bioinformatic analysis of a subset of B19V genomic sequences available in the NCBI nucleotide database ,representative of genotypes 1 to 3,a set of suitable universal primers and TaqMan probes was designed from the NS 1 gene of B19V.Aplasmid was used as a quantitative standard that contained the identical sequence of the B 19 target sequence .An internal control ( IC ) was included to prevent false negative results .Then,serial 1-log dilutions of quantitative standards were prepared and used in the qPCR assays for generation of a standard curve .Finally,the specificity,sensitivity and reproducibility of the assay were assessed.Results A linear relationship of the real-time PCR method for detecting B19V from 1 ×109copies/μl to 1 ×103 copies/μl was observed .The developed qPCR protocols allowed for the detection of genotypes 1 to 3 with a limit of detection ( LOD) of 10 copies/μl.Furthermore, the assay did not amplify other blood-borne viruses.The inter-and intra-assay variability analyses showed good reproducibility of the assay .Conclusion A universal real-time qPCR method for the detection of B19V DNA is established,which will facilitate the diagnosis of B19V infections and the screening of blood and plasma-derived products , thereby improving the viral safety of transfusion and plasma-derived products .

Objective To detect human parvovirus B19(B19V)DNA in source plasma pools and coagulation factor products and determine its prevalence and the level of contamination .Methods A pair of primers and a probe selected from the highly conserved sequences encoding the non-structural protein(NS1)of B19 were designed and synthesized.With the primer-probe combination ,source plasma pools and four types of coagulation factor products were determined for B 19V DNA by TaqMan real-time quantitative PCR.Results One-hundred and sixteen from 195 (59.49%) source plasma pools contained B19 DNA and concentrations up to 1.35 ×1010 copies/ml were measured.High frequencies of contamination were detected in factor Ⅷ (29 of 31; 93.55%), thrombin (10 of 10; 100%), fibrinogen (6 of 7; 85.71%) and prothrombin complex (8 of 9;88.89%).Conclusion These data show that B19V is a common contaminator in Chinese source plasma pools and coagulation factor products .Thus,B19V screening in Chinese source plasma seems desirable and significant for the safety of plasma derivatives in China .

Objective:To explore the effect of knowledge-clinical-sharing (KCS) model in humanistic care training of pediatric nurses.Methods:A total of 182 specialist nurses who were trained in Hunan Children's Hospital from June 2019 to December 2019 were selected as the research objects. Among them, 102 specialist nurses from June 2019 to August 2019 were selected as the control group, and 80 specialist nurses from September 2019 to December 2019 were selected as the intervention group. The Jefferson empathy scale was used to compare the effect of humanistic training before and after the implementation. SPSS 22.0 statistical software was used, measurement data were test by F-test and t-test, and the counting data were analyzed by Chi-square test. Results:After the implementation of humanistic care training based on KCS model, the empathy ability score of the intervention group was higher than that of the control group [(80.23±5.33) vs. (78.14±4.37)], and the difference was statistically significant ( P < 0.01). There was no significant difference between neonatal and pediatric specialist nurses before and after the training. Conclusion:The phased humanistic training based on KCS model can improve the empathy ability of pediatric nurses and enhance their confidence in the clinical implementation of humanistic care. However, the humanistic training mode of pediatric nurses should be improved according to the hospital's own situation, so as to adapt to the development of nursing industry.

Objective:To perform principal component analysis on the status quo of nurses' research ability, and explore the status quo of their relevant research needs.Methods:In this cross-sectional study, totally 588 clinical nurses working in a children's hospital in Hunan Province from July to December 2019 were selected by stratified random sampling and investigated with the self-designed questionnaire. The survey content included three parts: the first part was general information; the second part was the survey of the status quo of nurses' research ability; the third part was the survey of the status quo of nurses' research needs. A total of 588 questionnaires were distributed, and 577 questionnaires were collected, accounting for an effective recovery rate of 98.13%.Results:The three items with the higher scores in the survey of research ability status were frequently search for professional materials through the Internet (3.255±0.491) scores, able to perform statistical description (2.931±0.572) scores and able to use Chinese database (2.894±0.429) scores. The three items with the higher need for research ability were project application 80.94% (467/577) , paper writing 76.95% (444/577) and special lecture 71.92% (415/577) , whereas the three items with the lower need for research ability were continuing learning 46.10% (266/577) , retrieving documents 40.90% (236/577) and group symposium 38.30% (211/577) .Conclusions:Nursing staff has relatively good ability in information retrieval and basic statistics, but relatively poor ability in paper writing. The research ability of nurses can be summarized as statistical analysis ability, information retrieval/document reading ability, research topic selection ability and paper writing ability. Attention should be paid to whether there are high research needs in the weak links of research ability, and nursing research guidance and education should be provided purposively.