Objective To investigate the significance of hepatitis B virus (HBV),hepatitis C virus (HCV) and human immunodeficiency virus(HIV)nucleic acid detection in patients undergoing surgery/transfusion.Methods From October 2019 to June 2020,serological blood samples of HBV,HCV and HIV in patients undergoing surgery/transfusion were collected from 23 clinical medical institutions for HBV DNA,HCV RNA,HIV-1/2 RNA detection.The results of the two methods were compared and analyzed.Results A total of 10 173 samples were included in this study,in which HBV DNA positive in 1 142 cases (11.23%),HCV RNA positive in 34 cases (0.33%),HIV-1/2 RNA positive in 9 cases (0.09%),hepatitis B surface antigen (HBsAg) reactive in 987 cases (9.70%),HCV antibody (Anti-HCV) reactive in 77 cases (0.76%) and HIV antigen antibody (HIV Ag/Ab) reactive in 14 cases (0.14%).There was significant difference in HBV and HCV positive /reactive ratio between the two methods (P<0.001).The results of serological test and nucleic acid detection of the three viruses were consistent in 9 754 cases (95.88%) and inconsistent in 419 cases (4.12%).Among the inconsistent results,there were 263 cases of serological non-reaction and nucleic acid positive (HBV 254 cases,HCV 7 cases and HIV 2 cases),and 156 cases serological reaction and nucleic acid negative (HBV 99 cases,HCV 50 cases and HIV 7 cases).The consistency of serology and nucleic acid detection was very good for HBV (kappa=0.815),good for HIV (kappa=0.608),and fair for HCV (kappa=0.484).Compared with enzyme immunoassay method,the consistency between chemiluminescence method and nucleic acid detection for three viruses was better.Conclusion Nucleic acid detection is helpful to improve the detection rate and accuracy of HBV,HCV and HIV in patients undergoing surgery/transfusion.

Objective To analyze the inconsistent results between hepatitis C virus (HCV) serological test and nucleic acid test,and to investigate the significance of performing hepatitis C virus (HCV) nucleic acid test(NAT) in preoperative/pre-transfusion patients.Methods From October 2019 to June 2020,preoperative and pre-transfusion samples were collected for HCV serological and nucleic acid testing,and the inconsistent results between HCV serological testing and nucleic acid testing were analyzed.Results Among a total of 10 173 samples,57 cases are inconsistent inHCV serology and nucleic acid test results,including 50 cases of anti-HCV reactivity and HCV RNA negative(10 cases were treated with antiviral drugs and 5 cases were false positive),7 cases of anti-HCV nonreactivity and HCV RNA positive.A total of 8 regents were used for serological testing.5 cases of false positives were tested with reagent A.Compared to patients tested with regent A during antiviral therapy,the S/CO values of false positives samples were lower.The Ct values of 7 samples only HCV RNA positive were significantly higher than those of 27 samples both anti-HCV reactivity and HCV RNA positive (P<0.001).Conclusion For preoperative/pre-transfusion patients,nucleic acid testing can improve the accuracy of HCV testing and shorten the detection window in preoperative/pre-transfusion patients.

Objective To investigate the significance of nucleic acid test in judging the results of serological weak-reactive blood samples.Methods From October 1,2019 to June 30,2020 blood samples of patients before operation or transfusion were collected from 5 clinical medical institutions.The serological weak-reactive samples of HBV,HCV and HIV were detected by HBV DNA,HCV RNA,HIV-1/2 RNA test.Results Among the 5 782 samples,53 cases showed weak reaction,of which HBV positive in 24 cases,HCV positive in 25 cases and HIV positive in 4 cases.Further nucleic acid detection showed that 15 cases of HBV nucleic acid detection were reactive and 9 non-reactive;25 cases of HCV nucleic acid detection were non-reactive;4 cases of HIV nucleic acid detection were non-reactive.Conclusion The serological weak-reactiveis caused by high sensitivity of chemiluminescence method,which leads to a certain degree of false positive results.Nucleic acid detection canboth improvethe specificity and sensitivity,so as to ensure the accuracy of the results.

Objective To discuss the detection of hepatitis B virus (HBV) nucleic acid testing (NAT) in HBsAg-negative specimens.Methods A total of 10 173 results of HBV serology test and nucleic acid test from 23 hospitals in China between Oct 2019 and Jun 2020 were collected.HBsAg-negative specimens were Firstly included,and results of serum HBV DNA qualitative test were analyzed.Positive HBV DNA specimens were further included,and tested for serological marker HBsAb,HBeAg,HBeAb and HBcAb.The overall distribution of Ct value intervals and serological marker patterns were analyzed.Results 254(2.77%)of 9 186 HBsAg negative cases included were HBV DNA positive.There were eight serological patterns of HBV markers in 168 HBV DNA positive.166(98.81%) of those patients were occult hepatitis B infection(OBI).Conclusion HBV NAT could detect a certain percentage of HBV patients in HBsAg-negative specimens,especially for OBI patients.HBV NAT screening is important to detect OBI to avoid missing cases.

The function of red blood cell (RBC) oxygen supply involves the continuous process of external oxygen supply,the binding of oxygen to RBC,oxygen transport in the blood,oxygen release from RBC and oxygen uptake in tissue cells.Any factor affecting this process will influence the oxygen supply of RBC.Given the above process,indications for RBC transfusion and therapeutic efficacy should be evaluated in an overall dynamic view.

Objective To study the effct of gradient dilution of Japanese encephalitis virus (JEV) E protein antibody on the replication ability of dengue virus type Ⅱ(DENV-Ⅱ) in THP-1 cells.Methods Human THP-1 cells were co-infected with JEV-E antibodies of different dilution and DENV-Ⅱisolated from the serum of critically ill patients in Xishuangbanna,Yunnan Province.The standard curve of fluorescent quantitative PCR detection of DENV-Ⅱvirus was constructed.We used different dilution of JEV-E antibodies and DENV-Ⅱat different MOI (Multiplicity of infection) to infect THP-1 cells,then DENV-Ⅱvirus copy number was detected by real-time fluorescent quantitative PCR.Results We found 1/4 dilution degrees of JEV-E antibody concentration could promote DENV-Ⅱreplication in THP-1 cells,while high and low concentration of JEV-E antibody had no significant effect on DENV-Ⅱvirus replication in THP-1.Conclusion Flaviviruses JEV E protein antibody can induce antibody dependent enhanced infection of DENV-Ⅱin human THP-1 cells,which lays a theoretical foundation for the further study of antibody dependent enhanced infection among flaviviruses.

Objective To study the effect of riboflavin photochemical method on the function of lymphocyte in whole blood treated with a certain dose.Methods In the experimental group,the riboflavin solution was added to whole blood with a final concentration of 160 μmol/mL,then whole blood was injected into illumination blood bag.The thickness of whole blood was approximate 1.3 mm and the ultraviolet dose was 18 J/cm².The control group came from the same whole blood units without riboflavin and ultraviolet radiation.Lymphocytes in whole blood were separated to detect lymphocyte proliferation activity using CCK-8 kit and calculate proliferation inhibition rate.Stimulated by Phytohemagglutinin (PHA) with a final concentration of 1 μg/mL,cytokines secreted from lymphocytes were detected using cytokines kits.Results The lymphocyte proliferation inhibition rate was (74.18±14.00)% after riboflavin photochemical treatment.Cytokine IFN-γ,IL- 8,IL-12,IL-10,IL-13 and IL-4decreased by 63%,35%,61%,42%,77% and 8% respectively;After stimulated by PHA,cytokine IFN-γ,IL-8,IL-12,IL-10,IL-13,IL-4 decreased by 72%,43%,40%,78%,82% and 2% respectively;there was no statistical difference in the secretion of IL-4 between two groups.Conclusion Riboflavin photochemical treatment at a certain dose would inhibit partly the proliferation and cytokine secretion of lymphocytes in the whole blood.

Objective To explore the effect of LukS-PV on the proliferation in acute myeloid leukemia (AML) cells THP-1 and HL-60 by down-regulating methyltransferase SET8.Methods Real-time fluorescence quantitative PCR (qRT-PCR) and Western blot were used to detect the expression levels of SET8 mRNA and protein in THP-1 and HL-60 cells after treatment with different concentrations of LukS-PV.The lentiviral vector with knock-down SET8 expression was transfected into THP-1 and HL-60 cells.SET8 knockdown efficiency was detected by qRT-PCR and Western blot,and the effects of SET8 knock-down on cell proliferation was detected by EdU assay.Finally,on the basis of knocking down SET8,adding LukS-PV treatment for detecting cell proliferation by EdU to explore whether LukS-PV inhibits the proliferation of leukemic cells THP-1 and HL-60 by down-regulating SET8.Results In AML cell lines THP-1 and HL-60,the expression of SET8 and H4K20me1 in LukS-PV treated group was significantly lower than that in PBS control group in a concentration-dependent manner (P<0.05).Compared with the negative control group,SET8 expression level was significantly decreased in THP-1 and HL-60 cells transfected with knock-down SET8 lentiviral (P<0.05),and cell proliferation ability showed the same trend (P<0.05).Compared with the PBS control group,knocking down SET8 could inhibit cell proliferation (P<0.05),and the inhibition of cell proliferation was more obvious when LukS-PV was added on this basis (P<0.05).Conclusion Down-regulating SET8 inhibits the proliferation of THP-1 and HL-60 AML cells,and LukS-PV can inhibit cell proliferation by down-regulating SET8 and play an anti-leukemia effect.

Objective To explore the blood transfusion in maternal neonatal diseases and provideabetter clinical transfusion guidance.Methods Clinical data of 180 newborns treated with blood transfusion in Nanjing Maternity and Child Health Hospital from January 2018 to December 2019 were collected and analyzed retrospectively.Results 162 (90%) of 180 newborns treated with blood transfusion had maternal diseases.The majority of maternal blood transfusions were premature infants.The transfusion frequency was statistically significant for neonates with different gestational age and weight(P<0.05).The red blood cell transfusion frequency of premature and low birth weight infantswas positively correlated with the frequency and the volume of blood collection(P<0.05).Conclusion The proportion of maternal factors was higher in neonatal transfusions.The younger the gestational age and the lower the body weight of premature infants,the more cases and times of blood transfusion.

Objective To explore the effect of perioperative blood protection measures on the rapid rehabilitation of elderly patients undergoing total hip arthroplasty.Methods A total of 98 elderly patients undergoing total hip arthroplasty who were treated in our hospital from May 2016 to June 2019 were selected for study.Patients were divided into 2 groups by random number table method,with 49 patients in each group.In two groups,total hip arthroplasty was performed by the same group of medical staff,and the control group was administered perioperative management of routine rapid rehabilitation.The observation group was given perioperative blood protection measures based on this.Acute blood loss,transfusion rate,hemoglobin,hematocrit,red blood cell count,nursing satisfaction and complications were compared between two groups of patients.Results The amount of acute blood loss in the control group was higher than that in the observation group.The total blood transfusion rate in the control group was 16.33%,which was significantly higher than the observation group's 4.08% (P<0.05).There were no significant differences in hemoglobin,hematocrit,and red blood cell count between the two groups of patients before surgery(P>0.05).The hemoglobin,hematocrit,and red blood cell count in patients of the control group were significantly lower than those in the observation group(P<0.05).Patients in the observation group had higher nursing satisfaction scores than those in the control group,and the hospital stay was shorter than that in the control group,and the difference was statistically significant(P<0.05).The total observed incidence of complications in the control group was 24.49%,which was significantly higher than that of the observation group (8.16%) (P<0.05).Conclusion Compared with conventional rapid rehabilitation nursing,perioperative blood protection measures combined with rapid rehabilitation nursing can effectively reduce acute blood loss and blood transfusion rate,stabilize hemoglobin and hematocrit and other hematological indicators,improve patient satisfaction and shorten hospital stay.At the same time,it can also reduce the incidence of complications.

Objective To explore the solution of interference in cross matching of patients with multiple myeloma treated with CD38 monoclonal antibody and to formulate its transfusion strategy.Methods Multiple myeloma patients treated with the CD38 monoclonal antibody Daratumab used dithiothreitol (DTT) before and after treatment with antibody screening cells,spectral cells,and donor red blood cells.Unexpected antibody screening,identification and cross-matching were performed by microcolumn Gel method and polybrene method respectively.Results After chemotherapy with CD38 monoclonal antibody,ABO blood groups were consistent with forward and reverse typing in 5 patients.The microcolumn Gel method was positive for antibody screening,identification and forward cross-matching,while the polybrene method was negative.Unexpected antibody screening cells,spectrum cells and donor red cells were treated with 0.2mol/L DTT.The microcolumn Gel method was negative in antibody screening,identification and forward typing.After blood transfusion,the patients' hemoglobin increased significantly,but the serum bilirubin did not change obviously.Conclusion In patients with multiple myeloma treated with CD38 monoclonal antibody,red blood cells treated with 0.2mol/L DTT can be used for antibody screening and cross matching by microcolumn Gel,or direct polybrene,which can be equally effective.

Objective We conducted a retrospective study to evaluate the efficacy of intensified myeloablative conditioning regimen for unrelated donor single-unit umbilical cord blood transplantation (sUCBT) in acute myeloid leukemia with FLT3-ITD mutation.Methods We studied a total of 22 patients with FLT3-ITD positive acute myeloid leukemia undergoing sUCBT between Jun 2012 and Dec 2018 at hematology department, the first affiliated hospital of USTC(Anhui provincial hospital), and evaluated the therapeutic efficacy of intensified myeloablative conditioning regimen without antithymocyte globulin (ATG). Twelve males and 10 females were enrolled. Median age was 24 years (range 4-53) and median weight was 58 kg (range 15-77). Among them, 6 of the patients had NPM1 mutation. Twelve patients were in first complete remission (CR1), three in second complete remission (CR2), and seven in advanced-stage disease before transplantation. All patients received intensified myeloablative conditioning regimen. Four received total body irradiation/ cyclophosphamide/cytosine arabinoside (TBI/Cy/Ara-C). Fifteen received Busulfan/ cyclophosphamide/fludarabine(BU/Cy/Flu). Three received BU/Cy/Ara-C. Ciclosporin A (CsA) in combination with mycophenolate mofetil (MMF) without ATG was used approach to graft-versus-host disease (GVHD) prevention.Results Hematopoietic reconstitution had been achieved successfully in 20/22 patients. The cumulative incidence (CI) of neutrophil engraftment at 42 days was 90.9%, and the median time to engraftment was 17.5 days (range 11~25 days). The CI of platelet engraftment at 120 days was 84.6%, and the median time to engraftment was 37 days (range 20~68 days). Transplant-related complications were collected. Thirteen (59.1%) patients developed pre-engraftment syndrome (PES). Twelve had grade II-IV acute GVHD, and the CI was 54.5%. Six had grade III-IV acute GVHD, and the CI was 27.3%. Four had localized cutaneous chronic GVHD (cGVHD), and the CI at 720 days was 18.2%. No extensive cGVHD occurred. As of June 30, 2020, a median follow-up of 747.5 days（range 6-2 932). Three patients had primary recurrence. The 3-year CI of relapse was 13.6%. The 3-year overall survival and disease-free survival were 62.9% and 63.6% respectively.Conclusion The preliminary investigation of sUCBT with intensified myeloablative conditioning regimen for FLT3-ITD mutation acute myeloid leukemia showed good curative effect, and It is worthy to apply.

Objective To analyze the drug susceptibility,carbapenem resistance determinants and multi-locus sequence types of carbapenem resistant Escherichia coli.Methods Carbapenem-resistant E.coli isolates collected from our hospital were analyzed for the susceptibilities by Vitek 2 Compact.Detection of carbapenem resistance genes was performed by PCR and DNA sequencing,Multi-locus sequence typing was used to analyze the sequence types of bacteria.The transferability of bla_NDM gene was analyzed by conjugation experiment.Results Carbapenem-resistant E.coli were resistant to most of the commonly used antibiotics in clinical therapy.Nine out of 11 carbapenem resistant E.coli isolates were found to carry bla_NDM gene,and two isolates of them were found to carry bla_KPC gene.The results of multilocus sequence typing showed that 11 E.coli isolates were assigned into 7 sequence types (STs),which were ST167(n=3),ST6349(n=1),ST361(n=1),ST6335(n=1),ST617(n=1),ST7016(n=1) and ST354(n=1).The results of conjugation test showed that bla_NDM gene could be transferred,without bla_KPC gene being co-transferred.Conclusion The production of NDM is the main mechanism of E.coli conferring resistance to carbapenem,which is easy to spread among bacteria,infection control measures should therefore be further strengthened.

Objective To study the relationship between interleukin,TNF-αand lung function and immune function in children with bronchial asthma.Methods 178 children with bronchial asthma were divided into acute attack group(n=107) and remission group(n=71),and 83 healthy persons at the same period were enrolled in control group.The serum levels of interleukin [interleukin-2(IL-2),interleukin-4(IL-4)and interleukin-6(IL-6)],tumor necrosis factor-α(TNF-α),lung function related indicators [forced vital capacity(FVC),forced expiratory volume in the first second(FEV₁),FEV1/FVC and peak expiratory flow(PEF)] and immune function related indicators [T lymphocyte subsets(CD4⁺,CD8⁺andCD4⁺/CD8⁺)] were measured.Pearson correlation analysis was used to analyze the relationship between serum inflammatory factors and pulmonary function and immune function.Results The serum levels of IL-2,IL-4,IL-6 and TNF-αwere significantly increased in attack group(P<0.05).The levels of FVC,FEV1,FEV₁/FVC,PEF,CD4⁺and CD4⁺/CD8⁺ were significantly decreased in attack group(P<0.05).There was no significant difference in the levels of CD8⁺ among groups(P>0.05).Pearson correlation analysis showed that serum levels of IL-2,IL-4,IL-6 and TNF-αwere negatively correlated with FVC,FEV₁,FEV₁/FVC,PEF,CD4⁺,CD4⁺/CD8⁺ in attack group and remission group(P<0.05),but no with CD8⁺(P>0.05).Conclusion The levels of interleukin and TNF-αin children with bronchial asthma are significantly increased,while lung function and immune function are significantly reduced.Moreover,interleukin and TNF-αhave a negative correlation with lung function and immune function.The above indicators can be used to evaluate the therapeutic effect of bronchial asthma in children.

The purpose of red blood cell transfusion is to increase hemoglobin and improve hypoxia.The reasons that affect transfusion efficacy can be divided into immune factors and non-immune factors.Immune factors include the changes of red blood cells surface molecules,content of metabolites,the influence of immune substances in blood recipients and so on.Non-immune factors include storage damage of red blood cells,disease status of blood recipients and drug effects.Physicians are more concerned with transfusion-related adverse reactions but less with poor red blood cells transfusion responses.Here,we review the potential reasons for the poor responses of red blood cell transfusion,in order to provide theoretical support and new ideas for research and clinical practice.