A method based on QuEChERS coupled with ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was developed for the determination of seven kinds of fluorescent white agent residues in mushroom.After mixing with 10 mL of water, the sample was extracted with acidified acetonitrile, cleaned up by C18, primary secondary amine (PSA) and MgSO4.The separation of seven kinds of fluorescent white agents was performed on a C18 column with a gradient elution of acetonitrile and 0.1% acidified water as mobile phase.The target compounds were detected by electrospray ionization mass spectrometry in positive ion mode with multi reaction monitoring (MRM).Under the optimum conditions, the method had good linear relationship in the determination of the seven kinds of fluorescent white agents in a certain concentration range, with correlation coefficients greater than 0.991.Moreover, the limits of detection (S/N=3) were 0.05-0.4 μg/kg, the limits of quantitation (S/N=10) were 0.2-1.3 μg/kg, and the average recoveries for seven kinds of fluorescent white agent residues in msushroom were 70.1%-109.2%.In comparison with previous methods, the new procedure has characteristics of simple sample preparation and higher sensitivity.

Objective To express virus-like particles(VLP) of enterovirus 71 (EV71) in Han-senula polymorpha.Methods The coding sequences of P1 and 3CD genes of EV71 were optimized accord-ing to codon usage bias of Hansenula polymorpha for achieving high expression , and then cloned into the ex-pression vector PMV of Hansenula polymorpha .The recombinant expression vector PMV-P1-3CD was trans-formed into Hansenula polymorpha AU 0501 .The transformants were stably cultured in selective medium (Yeast Nitrogen Base) and screened for strains with positive P1 and 3CD genes by PCR.Then an induced cultivation on the recombinant strains were performed in a medium supplemented with methanol to a final concentration of 1.0%and the expressed products were analyzed by SDS-PAGE and Western blot assays to select high expression strains .The high expression strains were cultured in 30 L fermentor and its fermenta-tion products were analyzed by electronic microscope after purification .Results EV71 recombinant expres-sion strains were successfully constructed .The results of SDS-PAGE showed that the expressed products had obvious VP3, VP1, VP0 protein bands with molecular weights of 26×103, 33×103 and 35×103, respective-ly, which were consistent with the expected molecular weight of the fusion proteins .Western blot demonstra-ted that the expressed products could be specifically recognized by the polyclonal antibody against EV 71-VP1 at 33 ×10 3 , indicating its high immunoreactivity .ELISA confirmed that the expression level of EV 71 fermen-tation products was reached to 200 mg/L.Electronic microscope analysis showed that the VLP of recombi-nant EV71 were 24-30 nm in diameter with normal structure .Conclusion The virus-like particles of human enterovirus 71 are successfully expressed in Hansenula polymorpha , which provides a foundation for the fur-ther development of EV 71 VLP vaccine .

Objective:To do factor analysis related to post-stro ke anxiety.Method:206 inpatients(male 126,female 80,mean age 63?16 )with stroke were tested with SAS,DNF,MMSE,LES and ADL.Results:T h e rate of anxiety after 1 month of stroke in our sample was 18.4%(38 cases).Mu lt i-factorial analysis showed,many factors including female,younger at onset,sever ity of stroke,poor general physical condition,less compliance with treatment,and other psycho-social factors(heavier family burden,less income,more life events, less social support)were all associated with anxiety after stroke.Conchus ion:Anxiety is a common complication after stroke,its occurrence were asso ciated with a variety of factors including severity of illness and psychosocial factors.

OBJECTIVE:To conduct the quality assessment of artificial Aquilaria sinensis by"cutting inducing technique", and provide reference for its scientific planting and harvest. METHODS:GC-MC and HPLC were adopted to detect the volatile in-gredients,characteristic spectrum,incense tetraol and alcohol-soluble extract contents in 3 batches of artificial A. sinensis(Num. 1, 2,3,respectively for 5,10,20 years)by"cutting inducing technique". RESULTS:The volatile ingredients of 3 batches of artifi-cial A. sinensis mainly consisted of aromatic compounds,sesquiterpene compounds,fatty acid compounds and chromone com-pounds. The characteristic spectrums of samples 2,3 were basically the same with the reference substance of A. sinensis. The in-cense tetraol contents of 3 batches of samples were 0.078%-0.254%,and alcohol-soluble extract contents were 12.4%-20.8%. The characteristic spectrum and the incense tetraol content of sample 1 were not conformed to the standards in Chinese Pharmacopoeia (2015 edition,part 1). CONCLUSIONS:Artificial A. sinensis by"cutting inducing technique"shows similar volatile ingredients to natural A. sinensis. The quality of artificial A. sinensis for more than 10 years is conformed to the standards in Chinese Pharmaco-poeia(2015 edition,part 1),which can be used as medicine,replacing the natural A. sinensis.

Objective:To evaluate the in vitro cross-neutralization of serum antibodies in human and mice immunized with inactivated SARS-CoV-2 vaccine against Delta and Beta variants. Methods:Human serum samples after a second and a third dose of inactivated SARS-CoV-2 vaccine and mouse serum samples after a two-dose vaccination were collected. The neutralizing antibodies in the samples against SARS-CoV-2 strains of prototype, Delta and Beta variants were detected using micro-neutralization assay in biosafety level Ⅲ laboratory. The seroconversion rates and geometric mean titers (GMTs) of antibodies were calculated.Results:The seroconversion rates of antibodies in human serum samples against different SARS-CoV-2 strains were all above 95%. After two-dose vaccination, the GMTs of neutralizing antibodies against the prototype, Delta and Beta strains were 109, 41 and 15, respectively. The GMTs decreased by 2.7 folds and 7.3 folds for the Delta and Beta variants as compared with the prototype strain. After the booster vaccination, the GMTs of neutralizing antibodies against the prototype, Delta and Beta strains were 446, 190 and 86, respectively. The GMTs of neutralizing antibodies against Delta and Beta variants decreased by 2.3 folds and 5.2 folds as compared with that against the prototype strain. The seroconversion rates of antibodies against different SARS-CoV-2 strains in mouse serum samples were all 100%. The GMTs of neutralizing antibodies against the prototype, Delta and Beta strains were 2 037, 862 and 408, respectively. The GMTs decreased by 2.4 folds and 5.0 folds for the Delta and Beta variants.Conclusions:Inactivated SARS-CoV-2 vaccine could induce a certain level of neutralizing antibodies against Delta and Beta variants in both human and mouse models. Moreover, a third dose of vaccine induced higher levels of neutralizing antibodies against Delta and Beta variants in human. This study provided valuable data for the clinical application and protective evaluation of the inactivated SARS-CoV-2 vaccine.

Objective To investigate the expressions of transforming growth factor β1 (TGF-β1) and brain natrium peptide (BNP) in patients with diastolic heart failure (DHF),and to explore the correlation between plasma levels of TGF-β1,BNP and TGF-β1/BNP with parameter of diastolic function,diastolic dysfunction and New York Heart Association (NYHA) classification of cardiac function.Methods Hospitalized patients with DHF from October 2016 to November 2017 in Beijing Chaoyang Hospital were selected as subjects.At the same time,the age-and gender-matched non-heart failure hospitalized patients were selected as the control.The diastolic function index (E/e') was measured using cardiac ultrasound spectral Doppler and tissue Doppler methods.The diastolic dysfunction classification was evaluated according to the American Society of Echocardiography guidelines.Cardiac function was evaluated with NYHA classification.The levels of plasma TGF-β1 and BNP were measured with method of enzyme linked immunosorbent assay (ELISA).The correlation between the indicators was analyzed and the receiver operating characteristic (ROC) curve was drawn.Results A total of 186 patients were enrolled,including 114 patients as DHF group [54 males and 60 females,mean age (70.75 ± 11.45) years old] and 72 cases as control group [41 males and 31 females,mean age (68.74 ± 10.86) years old].The levels of TGF-β1 [(77.68 ± 42.31) ng/L] and BNP [(1 153.84 ± 564.96) ng/L] in patients with DHF were significantly higher than those of the control group [(18.76 ± 13.70),(264.07 ± 179.43) ng/L,t =15.62,13.77,P ＜ 0.01].Pearson correlation analysis showed that level of plasma TGF-β1 had a significant liner correlation with index E/e' (r =0.582,P ＜ 0.01),level of plasma BNP had a low-degree liner correlation with index E/e' (r =0.261,P ＜ 0.01),and TGF-β1/BNP had no correlation with index E/e' (r =0.081,P ＞ 0.05).Spearman correlation analysis showed that the levels of TGF-β1 and BNP were significantly correlated with diastolic dysfunction grading (r =0.473,0.417,P ＜ 0.01),while TGF-β1/BNP had no correlation with diastolic dysfunction grading (r =0.062,P ＞ 0.05).Plasma TGF-β1 and BNP had low-degree correlation with NYHA classification of heart failure (r =0.309,0.326,P ＜ 0.01),TGF-β1/BNP had no correlation with NYHA classification of heart failure (r =0.011,P ＞ 0.05).Logistic analysis showed that both plasma TGF-β1 and BNP were independent predictors of DHF (OR =1.264,2.283,P ＜ 0.05 or ＜ 0.01).The area under ROC curve (AUC) of BNP for prediction of DHF was 0.937 ± 0.064,and TGF-β1 was 0.597 ± 0.042.AUC areas of BNP and TGF-β1 were significantly different (P ＜ 0.01).Conclusions The expressions of plasma TGF-β1 and BNP in patients with DHF are higher than those without DHF.The levels of plasma TGF-β1 and BNP are significantly correlated with index E/e',diastolic dysfunction grading and NYHA classification.Both elevated BNP and TGF-β1 levels are independent predictors of DHF.Both plasma BNP and TGF-β1 have auxiliary diagnostic value on DHF and the diagnostic value of plasma BNP is greater than plasma TGF-β1.

Objective To experimentally verify the precision and accuracy of determining tritiated water in ambient air using the desiccant adsorption sampling–high temperature negative pressure desorption of liquid water (containing HTO)–liquid Scintillation counter method, and to provide technical support for developing standard methods for monitoring tritiated water in ambient air. Methods The relative standard deviation and recovery of multi-group samples were verified by collecting, testing, and analyzing environmental samples with different activity concentrations. The uncertainty of the method was evaluated, the main uncertainty components were identified, and the reliability of measurement results was analyzed. Through experimental comparison of different methods, the differences in the test results of different methods were examined. Results The relative standard deviation of multiple samples ranged from 6.7% to 7.9%, the recovery ranged from 95.7% to 97.3%, and the uncertainty was greatly affected by the sample counting rate, with no significant difference as compared to condensation sampling method. Conclusion The precision and accuracy of this method meet the requirements of environmental authorities for monitoring tritiated water in ambient air, and it can be widely used in the monitoring of tritiated water in ambient air.