Objective:To investigate the levels of diphtheria-specific binding antibodies and neutralizing antibodies in mice immunized with pneumococcal polysaccharide conjugate vaccine using diphtheria toxoid as a carrier.Methods:NIH mice were immunized with one batch of diphtheria, tetanus and acellular pertussis combined vaccine, absorbed (DTaP-1) or three different batches of 13-valent pneumococcal polysaccharide conjugate vaccine (PCV13, containing diphtheria toxoid vector) at three dilutions (5-, 10- and 20-fold dilution). Serum samples were collected to test for diphtheria-specific antibody titers and diphtheria potencies of the vaccines. Another three batches of DTaP vaccine and three batches of tetanus toxoid, reduced diphtheria toxoid and acellular pertussis combined vaccine (Tdap) were used to immunize NIH mice. Serum samples were collected and the diphtheria potencies were detected. Statistical analysis was performed using one-way analysis of variance.Results:At the 5-fold and 10-fold dilutions, the titers of diphtheria-specific antibodies induced by three batches of PCV13 vaccine were all lower than those by DTaP-1 vaccine (all P<0.001), while there was no statistically significant difference at the 20-fold dilution ( P>0.05). The diphtheria potencies of the DTaP-1 vaccine and the three batches of PCV13 vaccine were 100.5, 76.2, 64.5, and 62.0 IU/ml, respectively. The diphtheria potencies of another three batches of DTaP vaccine were 82.5, 83.6, and 79.9 IU/ml, respectively, and those of three batches of Tdap vaccine were 10.3, 12.2, and 12.9 IU/ml, respectively. There was no statistically significant difference in diphtheria potency between DTaP vaccine and PCV13 vaccine( P>0.05), while there was a statistically significant difference between Tdap vaccine and the PCV13 vaccine ( P<0.001). Conclusions:The pneumococcal polysaccharide conjugate vaccine with diphtheria toxoid has good diphtheria immunogenicity and can induce the production of higher levels of diphtheria-specific binding antibodies and protective neutralizing antibodies in vivo. Pneumococcal capsular polysaccharide exerts an immune enhancement effect on diphtheria toxoid. The relevant results provide valuable guidance for determining carrier protein dosage in bacterial polysaccharide conjugate vaccines, planning vaccine co-administration, and selecting the dosage of diphtheria toxoid antigen in the research and development of combined vaccines.

AIM:The current study aims to investigate the protective effect of farrerol against sepsis-induced cardiomyopathy and its mechanisms in mice.METHODS:Eighteen male C57BL/6 mice were randomly divided into the control,lipopolysaccharide(LPS;10 mg/kg),and LPS+farrerol(40 mg/kg)groups,with 6 mice in each group.The car-diac function of the mice was assessed via ultrasonography.Myocardial pathological changes and mitochondrial damage were examined via hematoxylin and eosin staining and transmission electron microscopy,respectively.TUNEL staining was performed to evaluate myocardial apoptosis,and RT-qPCR and ELISA were conducted to assess cardiac tissue inflam-matory factor mRNA expression and the serum inflammatory factor levels.Furthermore,the malondialdehyde(MDA),glutathione(GSH),superoxide dismutase(SOD),Fe2+,and reactive oxygen species(ROS)levels in the myocardial tis-sues were evaluated using kits and immunofluorescence.Western blot analysis was performed to investigate the levels of key proteins associated with apoptosis,ferroptosis,and the nuclear factor E2-related facor 2(Nrf2)/heme oxygenase-1(HO-1)signaling pathway.RESULTS:Farrerol pretreatment prevented the reduction of cardiac function in mice with SIC(P<0.01).Further,it decreased myocardial tissue pathological damage,mitochondrial ultrastructural disruption,the inflammatory factor levels(P<0.01),cardiac oxidative stress and Fe2+content(P<0.01),the expression of apoptotic cardiomyocytes(P<0.01),and the level of Bax expression(P<0.01).Finally,it increased the protein expression of Bcl-2,glutathione peroxidase 4(GPX4),solute carrier family 7 member 11(SLC7A11),Nrf2,and HO-1(P<0.05 or P<0.01).The control,LPS,and LPS+farrerol groups did not significantly differ in terms of left ventricular anterior wall end-diastolic depth and left ventricular posterior wall end-diastolic depth(P>0.05).CONCLUSION:Farrerol reduces LPS-induced septic myocardial injury and inhibits the development of ferroptosis in the myocardial tissues of mice,which may be related to the Nrf2/HO-1 signaling pathway.

AIM:The current study aims to investigate the protective effect of farrerol against sepsis-induced cardiomyopathy and its mechanisms in mice.METHODS:Eighteen male C57BL/6 mice were randomly divided into the control,lipopolysaccharide(LPS;10 mg/kg),and LPS+farrerol(40 mg/kg)groups,with 6 mice in each group.The car-diac function of the mice was assessed via ultrasonography.Myocardial pathological changes and mitochondrial damage were examined via hematoxylin and eosin staining and transmission electron microscopy,respectively.TUNEL staining was performed to evaluate myocardial apoptosis,and RT-qPCR and ELISA were conducted to assess cardiac tissue inflam-matory factor mRNA expression and the serum inflammatory factor levels.Furthermore,the malondialdehyde(MDA),glutathione(GSH),superoxide dismutase(SOD),Fe2+,and reactive oxygen species(ROS)levels in the myocardial tis-sues were evaluated using kits and immunofluorescence.Western blot analysis was performed to investigate the levels of key proteins associated with apoptosis,ferroptosis,and the nuclear factor E2-related facor 2(Nrf2)/heme oxygenase-1(HO-1)signaling pathway.RESULTS:Farrerol pretreatment prevented the reduction of cardiac function in mice with SIC(P<0.01).Further,it decreased myocardial tissue pathological damage,mitochondrial ultrastructural disruption,the inflammatory factor levels(P<0.01),cardiac oxidative stress and Fe2+content(P<0.01),the expression of apoptotic cardiomyocytes(P<0.01),and the level of Bax expression(P<0.01).Finally,it increased the protein expression of Bcl-2,glutathione peroxidase 4(GPX4),solute carrier family 7 member 11(SLC7A11),Nrf2,and HO-1(P<0.05 or P<0.01).The control,LPS,and LPS+farrerol groups did not significantly differ in terms of left ventricular anterior wall end-diastolic depth and left ventricular posterior wall end-diastolic depth(P>0.05).CONCLUSION:Farrerol reduces LPS-induced septic myocardial injury and inhibits the development of ferroptosis in the myocardial tissues of mice,which may be related to the Nrf2/HO-1 signaling pathway.

Objective:To investigate the levels of diphtheria-specific binding antibodies and neutralizing antibodies in mice immunized with pneumococcal polysaccharide conjugate vaccine using diphtheria toxoid as a carrier.Methods:NIH mice were immunized with one batch of diphtheria, tetanus and acellular pertussis combined vaccine, absorbed (DTaP-1) or three different batches of 13-valent pneumococcal polysaccharide conjugate vaccine (PCV13, containing diphtheria toxoid vector) at three dilutions (5-, 10- and 20-fold dilution). Serum samples were collected to test for diphtheria-specific antibody titers and diphtheria potencies of the vaccines. Another three batches of DTaP vaccine and three batches of tetanus toxoid, reduced diphtheria toxoid and acellular pertussis combined vaccine (Tdap) were used to immunize NIH mice. Serum samples were collected and the diphtheria potencies were detected. Statistical analysis was performed using one-way analysis of variance.Results:At the 5-fold and 10-fold dilutions, the titers of diphtheria-specific antibodies induced by three batches of PCV13 vaccine were all lower than those by DTaP-1 vaccine (all P<0.001), while there was no statistically significant difference at the 20-fold dilution ( P>0.05). The diphtheria potencies of the DTaP-1 vaccine and the three batches of PCV13 vaccine were 100.5, 76.2, 64.5, and 62.0 IU/ml, respectively. The diphtheria potencies of another three batches of DTaP vaccine were 82.5, 83.6, and 79.9 IU/ml, respectively, and those of three batches of Tdap vaccine were 10.3, 12.2, and 12.9 IU/ml, respectively. There was no statistically significant difference in diphtheria potency between DTaP vaccine and PCV13 vaccine( P>0.05), while there was a statistically significant difference between Tdap vaccine and the PCV13 vaccine ( P<0.001). Conclusions:The pneumococcal polysaccharide conjugate vaccine with diphtheria toxoid has good diphtheria immunogenicity and can induce the production of higher levels of diphtheria-specific binding antibodies and protective neutralizing antibodies in vivo. Pneumococcal capsular polysaccharide exerts an immune enhancement effect on diphtheria toxoid. The relevant results provide valuable guidance for determining carrier protein dosage in bacterial polysaccharide conjugate vaccines, planning vaccine co-administration, and selecting the dosage of diphtheria toxoid antigen in the research and development of combined vaccines.

COVID-19 is caused by a novel coronavirus-severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2), which has being spreading around the world, posing a serious threat to human health and lives. Neutralizing antibodies and small molecule inhibitors for virus replication cycle are the main antiviral treatment for novel coronavirus recommended in China. To further promote the rational use of antiviral therapy in clinical practice, the National Center for Infectious Diseases (Beijing Ditan Hospital Capital Medical University and the First Affiliated Hospital, Zhejiang University School of Medicine) invited experts in fields of infectious diseases, respiratory and intensive care to develop an Expert Consensus on Antiviral Therapy of COVID-19 based on the Diagnosis and Treatment Guideline for COVID-19 ( trial version 10) and experiences in the diagnosis and treatment of COVID-19 in China. The consensus is concise, practical and highly operable, hopefully it would improve the understanding of antiviral therapy for clinicians and provide suggestions for standardized medication in treatment of COVID-19.

Objective To establish a new method for rapid detection of β-thalassemia by investigating six clinical common mutation types.Methods Fifty cases of clinical wild-type samples and 42 cases ofβ-thalassemia samples were collected, and β-globin gene was amplified by PCR.Uniform ligation probe ( ULP) specific probes were designed for hybridization reaction to increase the reaction specificity and real-time PCR was performed to increase the sensitivity.After that, PCR products were verified by agarose electrophoresis.After examining the specificity and sensitivity, Kappa test between LDR-ULP method and reverse dot blot( RDB) method was conducted.Results Hybridization efficiency was improved 2.53 times by LDR-ULP hybridization.Each mutant type showed a significant amplification curve, whereas the wild-type had no significant curve within 40 cycles.The limit of determination of this method was 5 pg.The results of 92 cases of peripheral blood samples detected by the method of LDR-ULP and RDB were completely consistent.Conclusion In this study, a simple, inexpensive, rapid new method to detect β-thalassemia were established.