Objective:To evaluate the efficacy and prognostic factors of allogeneic hematopoietic stem cell transplantation (allo-HSCT) in patients with secondary acute myeloid leukemia (sAML) .Methods:In this multicenter, retrospective clinical study, adult patients aged ≥18 years who underwent allo-HSCT for sAML at four centers of the Zhejiang Hematopoietic Stem Cell Transplantation Collaborative Group from January 2014 to November 2022 were included, and the efficacy and prognostic factors of allo-HSCT were analyzed.Results:A total of 95 patients were enrolled; 66 (69.5%) had myelodysplastic syndrome-acute myeloid leukemia (MDS-AML) , 4 (4.2%) had MDS/MPN-AML, and 25 (26.3%) had therapy-related AML (tAML) . The 3-year CIR, LFS, and overall survival (OS) rates were 18.6% (95% CI 10.2%-27.0%) , 70.6% (95% CI 60.8%-80.4%) , and 73.3% (95% CI 63.9%-82.7%) , respectively. The 3-year CIRs of the M-AML group (including MDS-AML and MDS/MPN-AML) and the tAML group were 20.0% and 16.4%, respectively ( P=0.430) . The 3-year LFSs were 68.3% and 75.4%, respectively ( P=0.176) . The 3-year OS rates were 69.7% and 75.4%, respectively ( P=0.233) . The 3-year CIRs of the groups with and without TP53 mutations were 60.0% and 13.7%, respectively ( P=0.003) ; the 3-year LFSs were 20.0% and 76.5%, respectively ( P=0.002) ; and the 3-year OS rates were 40.0% and 77.6%, respectively ( P=0.002) . According to European LeukmiaNet 2022 (ELN2022) risk stratification, the 3-year CIRs of patients in the low-, intermediate-, and high-risk groups were 8.3%, 17.8%, and 22.6%, respectively ( P=0.639) . The three-year LFSs were 91.7%, 69.5%, and 65.6%, respectively ( P=0.268) . The 3-year OS rates were 91.7%, 71.4%, and 70.1%, respectively ( P=0.314) . Multivariate analysis revealed that advanced disease at allo-HSCT and TP53 mutations were independent risk factors for CIR, LFS, and OS. Conclusion:There was no significant difference in the prognosis of patients who underwent allo-HSCT among the MDS-AML, MDS/MPN-AML, and tAML groups. Advanced disease at transplantation and TP53 mutations were poor prognostic factors. ELN2022 risk stratification had limited value for predicting the prognosis of patients with sAML following allo-HSCT.

Objective:To obtain hepatitis B virus capsid-like particles (CLPs) displaying B-cell linear epitopes of SARS-CoV-2 spike protein and evaluate their immunogenicity.Methods:Four recombinant plasmids expressing fusion proteins (M1-HBc, S1-57-HBc, S14P5-HBc and S21P2-HBc) were constructed by separately replacing codon of alanine at position 80 of hepatitis B virus core protein (HBc) with four genes coding for four B-cell linear epitopes (M1, S1-57, S14P5 and S21P2). These four recombinant proteins were expressed in E. coli BL21 Star (DE3) strains. The expression products were identified using SDS-PAGE, Western blot and native agarose gel electrophoresis (NAGE). CLPs were purified by sucrose density gradient ultracentrifugation, verified for antigenicity by Western blot and used to immunize BALB/c mice. Serum antibody titers were detected by ELISA. Results:The recombinant fusion proteins M1-HBc and S1-57-HBc self-assembled into M1-CLP and S1-57-CLP. The titer of antibody against S1-57 polypeptide in S1-57-CLP-immunized mouse serum approached 1∶1 000 000.Conclusions:Hepatitis B virus CLPs displaying SARS-CoV-2 M1 or S1-57 linear epitopes are successfully expressed in a prokaryotic system and purified. S1-57-CLP has good immunogenicity. This study provides a new idea for the development of novel diagnostic reagents and vaccines for SARS-CoV-2.

Objective To detect the expression level of 4-aminobutyrate aminotransferase(ABAT)in bone marrow of patients with myelodysplastic syndrome(MDS),and analyze its influence on clinicopathological features and prognosis of patients.Methods From January 2016 to March 2020,92 patients with MDS and 30 patients with acute myeloid leukemia(AML)from the First Affiliated Hospital of Xingtai Medical College were retrospectively collected.Meanwhile,30 patients with immunothrombocytopenia who did not develop MDS or other clonal diseases of the blood system during a 3-year follow-up were collected as control group.Real-time quantitative fluorescent PCR(qRT-PCR)was used to detect the relative expression level and methylation level of ABAT mRNA of all patients,and the relative expression level and methylation level of ABAT mRNA among different clinical characteristics of MDS patients were compared.Multivariate logistic regression analysis was used to analyze the risk factors affecting the adverse prognosis of MDS.The clinical value of detecting ABAT methylation level in predicting poor prognosis of MDS patients was analyzed by receiver operating characteristic(ROC)curve.Kaplan-Meier method was used to calculate the 3-year survival rate between groups with different ABAT mRNA relative expression levels and methylation levels,and log-rank test was used for their comparison.Results The expression level of ABAT mRNA in MDS group(0.42±0.08)was lower than that in control group(0.56±0.15)and AML group(0.52±0.10),while the methylation level of ABAT(32.51±5.32)was higher than that of AML group(26.21±4.58)and control group(10.25±4.31),and the differences were significant(t=4.251,4.562;10.415,8.326,all P<0.001).The methylation level of ABAT in high-risk patients(42.65±5.32)was higher than that in low-risk patients(25.63±4.16),intermediate-risk-1 patients(30.59±2.51)and intermediate-risk-2 patients(33.25±3.69)by IPSS risk grade,and the differences were significant(t=8.329,7.077,15.874,all P<0.001).Poor Karyotype analysis result[OR(95%CI):4.973(1.524～8.581),P=0.004],high IPSS risk grade[OR(95%CI):8.542(2.365～14.521),P<0.001]and ABAT hypermethylation level[OR(95%CI):6.178(1.589～13.021),P<0.001]were the risk factors affecting the poor prognosis of MDS.The cut-offvalue of ABAT methylation level to predict the poor prognosis of MDS were 30.54,and the area under the curve(AUC),the sensitivity and specificity were 0.92,0.874 and 0.851,respectively.The 3-year survival rate of the high ABAT methylation group(>30.54)was 66.67%,which was lower than that of the low ABAT methylation group(≤30.54)was 93.18%,with significant difference(Log-rank x2=9.814,P=0.002).Conclusion The ABAT methylation levels in MDS bone marrow increase,which is a risk factor affecting the poor prognosis of patients.ABAT basal level>30.54 is expected to become a factors predicting the poor prognosis of patients.

Animals ; SARS-CoV-2 ; Antibodies, Neutralizing ; Macaca mulatta ; COVID-19/prevention & control* ; Antibodies, Viral ; Vaccines

Objective: To describe the characteristics of clinical manifestations and epidemiology of children with 2019 novel coronavirus (2019-nCoV) infection. Methods: All 34 children with laboratory-confirmed 2019-nCoV infection by quantitative real-time reverse transcription-PCR through nasopharyngeal swab specimens were admitted to the Third People’s Hospital of Shenzhen from January 19 to Febuary 7, 2020. Clinical data and epidemiological history of these patients were retrospectively collected and analyzed. Results: Among the 34 cases, 14 were males, and 20 were females. The median age was 8 years and 11 months. No patients had underlying diseases. There were 28 children (82%) related with a family cluster outbreak. There were 26 children (76%) with a travel or residence history in Hubei Province. These patients could be categorized into different clinical types, including 22 (65%) common cases, 9 (26%) mild cases and 3 (8.8%) asymptomatic cases. No severe or critical cases were identified. The most common symptoms were fever (17 cases, 50%) and cough (13 cases, 38% ). In the 34 cases, the white blood cell counts of 28 cases (82%) were normal. Five cases had white blood cell counts more than 10×10⁹/L. One case had white blood cell counts less than 4×10⁹/L. Neutropenia and lymphopenia was found in one case, respectively. C-reactive protein levels and erythrocyte sedimentation rates were elevated in 1 and 5 case, respectively. Elevated procalcitonin was found in 1 case and D-Dimer in 3 cases. The levels of lactic dehydrogenase (LDH) were more than 400 U/L in 10 cases. The CT images of these patients showed bilateral multiple patchy or nodular ground-glass opacities and/or infiltrating shadows in middle and outer zone of the lung or under the pleura. Twenty patients were treated with lopinavir and ritonavir. Glucocorticoids and immunoglobulin were not used in any cases. All the cases improved and were discharged from hospital. Further following up was need. Conclusions The clinical manifestations in children with 2019-nCoV infection are non-specific and are milder than that in adults. Chest CT scanning is heplful for early diagnosis. Children's infection is mainly caused by family cluster outbreak and imported cases. Family daily prevention is the main way to prevent 2019-nCoV infection.

Objective:To compare the evaluation ability and effect of the nursing delirium screening scale (Nu-DESC) and the intensive care delirium screening checklist (ICDSC) on the patients with delirium after percutaneous coronary intervention(PCI).Methods:A total of 128 patients who were admitted to cardiovascular surgery from June 2018 to June 2019 for PCI intervention were selected by convenient sampling method.Data were collected by general data questionnaire, restless sedation scale (RASS), Nu-DESC and ICDSC.SPSS 18.0 was used to analyze the collected data.The patients' receiver operating characteristic(ROC) curve and area under the ROC curve(AUC) were used to analyze Nu-DESC scale and ICDSC scale to evaluate the delirium ability of patients after PCI. Kappa value was used to test the consistency of the scale.Bayes discriminant analysis was used to analyze the accuracy of Nu-DESC and ICDSC in judging delirium after PCI.Results:The AUC of Nu-DESC and ICDSC was 0.902 and 0.857 respectively, and the difference between them was AUC=0.045 ( Z=3.489, P<0.001); the best critical value of Nu-DESC and ICDSC for delirium evaluation was 2 and 4. The cross validation accuracy of Nu-DESC and ICDSC for delirium determination was 87.5% and 83.6%, respectively.The delirium rates of DSM-Ⅳ, Nu-DESC and ICDSC were 33.6%, 34.4% and 43.7%, respectively. Conclusion:Compared with ICDSC, Nu-DESC is more suitable for the evaluation of delirium after PCI.

Objective :To explore therapeutic effect of percutaneous coronary intervention (PCI ) combined hydro‐chloride tirofiban on aged patients with acute coronary syndrome (ACS) complicated diabetes mellitus (DM) and its safety .Methods :A total of 218 ACS + DM patients were selected .According to randomization method ,they were divided into routine treatment group (n＝107 ,received standard PCI) and tirofiban group (n＝111 ,received tirofi‐ban injection in coronary artery based on routine treatment group ).Operation indexes ,incidence rates of hemor‐rhage during hospitalization and major adverse cardiovascular events (MACE) within six months after PCI were ob‐served and compared between two groups .Results : There was no significant difference in door‐to‐balloon time , number and length of implanted stents between two groups , P＞0.05 all.Compared with routine treatment group , there were significant rise in percentages of TIMI grade 3 blood flow (75. 70% vs.91.89%) ,MBG grade 2 ～3 (69.16% vs.85.56%) and ST segment regression ＞50% within 90min after PCI (77. 57% vs.92.79%) ,and sig‐nificant reduction in corrected TIMI frame count (CTFC)[(33. 05 ± 8.37) frames vs.(26.54 ± 5.47) frames]in tiro‐fiban group , P＜0.01 all.There was no significant difference in incidence rate of hemorrhage events during hospi‐talization between two groups , P＝0.375. Incidence rate of MACE within six months in tirofiban group was signifi‐cantly lower than that of routine treatment group ,P＝0.001 .Conclusion :Intracoronary tirofiban injection based on routine PCI can significantly improve postoperative myocardial perfusion level ,reduce incidence rate of short‐term MACE after PCI without increasing incidence rate of hemorrhage in aged ACS patients .Its safety is good .

We developed a pre-treatment method to remove interfering substances during quantification of 146S antigens in foot-and-mouth disease (FMD) vaccines by high performance size exclusion chromatography (HPSEC). Three methods, including ultracentrifugation, PEG precipitation and nuclease digestion, were optimized and compared for removal efficiency of the interfering impurities in FMD vaccines. Under optimized conditions, the 146S contents in two batches of FMD vaccines were determined to be 7.1 and 7.6 μg/mL by ultracentrifugation, 9.7 and 10.4 μg/mL by PEG precipitation, and 10.5 and 10.4 μg/mL by nuclease digestion. The optimal condition for nuclease digestion using Benzonase determined by response surface method was as follows: appending Benzonase into 200 μL of antigen phase to a final concentration of 421 U/mL and incubating at 25.1 °C for 1.29 h. This method has advantages including efficient removal of the interfering impurities, fast processing speed, and mild operating conditions. Then 12 bathes of FMD vaccines with different serotypes produced by 4 manufacturers were tested to verify the established treatment method. Results showed the method was applicable to various FMD vaccines with good reproducibility (RSD<5.3%, n=3). The developed method removed interference from impurities during quantification of 146S, and therefore would broaden the application of HPSEC in vaccine quality control and ensure the accuracy and reliability.
Animals ; Chromatography, Gel ; Foot-and-Mouth Disease ; Foot-and-Mouth Disease Virus ; Reproducibility of Results ; Viral Vaccines

The aim of this study is to quantify the 146S antigen in foot-and-mouth disease virus (FMDV) inactivated vaccine by size-exclusion chromatography (SEC). The analysis was performed on a TSKgel G4000SWXL column (7.8 mm×30 cm), with a pH 7.2 buffer salt system as the mobile phase. The flow rate was 0.6 mL/min, the injection volume was 100 μL and the detection wavelength was 259 nm. The calibration curve was established by using purified inactivated FMDV (serotype O) 146S antigen; 3 batches of vaccine formulated by inactivated antigen solution were tested to verify the accuracy, reproducibility, specificity and tolerability of the method. At last 16 batches of vaccine were determined by the SEC method. Results showed a good linearity between peak area and concentration of 146S antigen in the range between 0.56 and 67.42 μg/mL (R2=0.996, n=10), and the average recovery rate of 146S antigen in the 3 batches of vaccine formulated in lab were 93.6% (RSD=2.7%, n=3), 102.3% (RSD=2.6%, n=3), and 95.5% (RSD=5.1%, n=3). The method was proved accurate and reliable with good reproducibility (RSD=0.5%, n=6), and applied to determine 16 batches of the commercial FMDV vaccine. According to the above results, the SEC method is high effective for 146S antigen quantify in the inactivated FMDV vaccine and would provide strong support for the vaccine quality control.

Objective: To investigate the clinical features and change trend of patients with acute coronary syndrome(ACS) undergoing emergent percutaneous coronary intervention(PCI). Methods: In this retrospective study, we retrieved all medical records of 4 907 ACS patients who underwent emergent PCI in Fuwai hospital from January 1,2010 to December 31,2016. We analyzed the clinical features and change trend in these patients. According to clinical diagnosis, patients were grouped as ST-elevated myocardial infarction(STEMI) group (3 719 cases) and NSTE-ACS group (patients with non-STEMI and unstable angina, 1 188 cases). Results: The ACS patients were aged (59.5±11.8) years old. There were 3 772 males and 1 135 females. The annual number of ACS patients underwent emergent PCI increased from 412 patients in 2010 to 1 067 patients in 2016. The number of NSTE-ACS patients increased from 11.4% (47/412) in 2010 to 26.5% (283/1 067) in 2016. Compared with STEMI group, patients in NSTE-ACS group were significantly older ((61.2±10.9) years old vs. (58.9±12.1) years old,P<0.01).The percent of female patients (30.1% (358/1 188) vs. 20.9% (777/3 719), P < 0.01), history of hypertension (69.1% (821/1 188) vs. 60.4% (2 248/3 719,P <0.01), previous PCI (25.8% (307/1 188) vs. 12.4% (461/3 719), P <0.01), and previous coronary artery bypass grafting (3.0% (36/1 188) vs. 1.0% (37/3 719), P <0.01) were all significantly higher in NSTE-ACS group than in STEMI group. On the other hand, NSTE-ACS patients presented less chronic renal failure (2.9% (35/1 188) vs. 4.3% (173/3 719), P <0.05) and hepatic dysfunction (8.5% (101/1 188) vs. 13.3% (495/3 719), P<0.01) as compared to ACS patients. In coronary angiography, NSTE-ACS patients had a higher prevalence of left-main disease (14.0% (166/1 188) vs. 7.8% (291/3 719), P<0.012 5) and triple vessel disease (47.8% (568/1 188) vs. 43.5% (1 619/3 719), P<0.012 5). There were no differences in prevalence of diabetes mellitus (31.9% (1 187/3 719) vs. 34.8% (414/1 188),P>0.05) and acute renal failure (0.1% (38/3 719) vs. 0.6% (7/1 188),P>0.05) between STEMI group and NSTE-ACS group. Conclusions This single center retrospective analysis reveals that there is an increasing trend of NSTE-ACS patients from 2010 to 2016. Furthermore, there are more high-risk clinical characteristics in NSTE-ACS patients than in STEMI patients.