Broad humoral immunity generated in mice by a formulation composed of two antigens from the Delta variant of SARS-CoV-2.

Due to the rapid development of new variants of SARS-CoV-2 as well as the real threat of new coronavirus zoonosis events, the development of a preventive vaccine with a broader scope of functionality is highly desirable. Previously, we reported the functionality of a nasal formulation containing the nucleocapsid protein and the receptor-binding domain (RBD) of the spike protein of the Delta variant of SARS-CoV-2 combined with the ODN-39M adjuvant. This combination induced cross-reactive immunity in mucosal and systemic compartments at the sarbecovirus level. In the present study, we explored the magnitude of the immunity generated in BALB/c mice by the same formulation with alum added as an additional adjuvant, to enhance the humoral immunity against the two antigens. Animals were immunized with three doses of the bivalent formulation, administered by subcutaneous route. Humoral immunity was tested by ELISA, and the neutralizing capacity of the resulting antibodies (Abs) was evaluated using a surrogate test and a vesicular stomatitis virus (VSV) pseudovirus-based assay. Cell-mediated immunity was also investigated using an IFN-γ ELISpot assay. High levels of antibodies against both antigens (N and RBD) were obtained upon immunization. Anti-RBD Abs with neutralizing capacity reacted with the RBD of three SARS-CoV-2 variants tested, including Omicron. Abs recognizing the nucleocapsid proteins of SARS-CoV-1 and the SARS-CoV-2 Delta and Omicron variants were also detected. Taken together, these results suggest that this bivalent formulation could be an attractive component of a pancorona vaccine able to broaden the scope of humoral immunity against both antigens. This will be particularly important for the reinforcement of immunity in previously vaccinated and/or infected populations.

Wnt5a-Flt1 activation contributes to preterm altered cerebral angiogenesis after prenatal inflammation.

OBJECTIVE: Intraventricular hemorrhage (IVH) causes morbidity and mortality in preterm infants and prenatal exposure to inflammation contributes to brain injury. Moreover, prenatal exposure to severe inflammation increases the risk of IVH in preterm neonates. The current study investigated whether intrauterine exposure to inflammation affects cerebral angiogenesis and its underlying mechanisms. METHODS: Wnt5a, flt1, and vascular endothelial growth factor (VEGF)-A levels in cord blood serum (stored in a bio-bank) of the enrolled patients were measured via enzyme-linked immunosorbent assay. A preterm prenatal inflammation exposure model was established in rats by intraperitoneal injection intraperitoneally during pregnancy. Angiogenesis of cerebral tissue was analyzed using immunohistochemistry. Wnt5a, flt1, and VEGF-A expression levels were measured via immunohistochemistry, immunofluorescence, or western blotting. The correlation between Wnt5a and flt1 expression and the cerebral vessel area was also analyzed. RESULTS: The Wnt5a and flt1 levels in the cord blood serum were significantly higher in the amnionitis group than in the non-amnionitis group. The VEGF-A level in the cord blood serum was significantly lower in the amnionitis group. In the rat model, preterm rats in the prenatal inflammation group exhibited increased microglial cell infiltration and decreased vessel area and diameter in the cerebral tissue compared to the control group. Wnt5a was located in microglial cells, and Wnt5a and flt1 expression in brain tissue significantly increased after prenatal lipopolysaccharide (LPS) exposure. VEGF-A expression declined after prenatal LPS exposure. The cerebral vessel area was negatively correlated with Wnt5a and flt1 expression. CONCLUSION: Disordered cerebral angiogenesis is associated with increased Wnt5a-Flt1 activation in microglial cells after exposure to intrauterine inflammation.

Autologous cord blood mononuclear cell infusion for the prevention of bronchopulmonary dysplasia in very preterm monozygotic twins: A study protocol for a randomized, placebo-controlled, double-blinded multicenter trial.

Background: Preterm-associated complications remain the main cause of neonatal death. Survivors face the challenges of short- and long-term complications. Among all complications, bronchopulmonary dysplasia (BPD) remains the first important cause of neonatal mortality and morbidity. Current treatment does not address this main preterm complication. Cord blood is regarded as a convenient source of stem cells. The paracrine bioactive factors of stem cells contribute to tissue repair and immune modulation. Our clinical studies and those of others have shown that cord blood cell infusion is both safe and possibly effective in the prevention and treatment of BPD. The therapeutic use of cord blood has emerged as a promising therapy. However, the genetic heterogeneity between control and intervention groups may reduce the comparability especially among small sample trials. The purpose of this study protocol is to investigate the effects of autologous cord blood mononuclear cell (ACBMNC) infusion on the prevention of BPD in very preterm monozygotic twins of less than 32 gestation weeks. Methods: In this prospective, randomized, placebo-controlled, double-blinded multicenter clinical trial, 60 pairs of monozygotic twin preterm neonates of less than 32 weeks admitted to the Neonatal Intensive Care Unit are randomly assigned to receive intravenous ACBMNC infusion (targeted at 5 × 107 cells/kg) or placebo (normal saline) within 24 h after birth in a 1:1 ratio. The primary outcome will be survival without BPD at 36 weeks of postmenstrual age. The secondary outcomes will include the mortality rate, BPD severity, other common preterm complication rates, respiratory support duration, length and cost of hospitalization, and long-term respiratory and neurodevelopmental outcomes during a 2-year follow-up. Furthermore, we will perform single-cell RNA sequencing for cord blood cells and blood cells 3-10 days after intervention and detect whether reactive oxygen species and inflammatory cytokines are present. Conclusion: This will be the first randomized, placebo-controlled, double-blinded trial to evaluate the efficacy of ACBMNC infusion to prevent BPD in monozygotic twin premature infants and investigate the underlying protective mechanisms. The results of this trial will provide valuable clinical evidence for translational application of cord blood cell therapy in very preterm infants.Trial registration: ClinicalTrials.gov, NCT05087498, registered 10/09/2021, https://register.clinicaltrials.gov/prs/app/action/SelectProtocol?sid=S000BAD7&selectaction=Edit&uid=U0002PLA&ts=2&cx=qvyylv.

Simultaneous Determination of 6 Residual Organic Solvents in Xingnaojing Injection by Headspace GC / 中国药房

OBJECTIVE:To establish the method for simultaneous determination of 6 residual organic solvents in Xingnaojing injection,such as methanol,ethanol,isopropanol,n-butanol,ethyl acetate and acetonitrile.METHODS:Headspace GC method was adopted.The determination was performed on DB-624 capillary column by temperature programming with the injector temperature of 200 ℃;flame ionization detector was adopted with the temperature of 250 ℃;carrier gas was nitrogen with flow rate of 25 mL/min and split ratio of 35 ∶ 1;headspace sampling size was 1 mL,and heating temperature of headspace sampling was 80 ℃;equilibrium time was 15 min.RESULTS:The linear ranges of methanol,ethanol,isopropanol,n-butanol,ethyl acetate and acetonitrile were 15.00-240.00 μg/mL (r =0.999 9),25.00-400.00 μg/mL (r =0.999 9),25.00-400.00 μg/mL (r =0.999 9),25.00-399.99 μg/mL(r=0.999 9),25.00-399.99 μg/mL(r=0.999 8) and 5.00-80.00 μg/mL(r=0.999 9).The LOQ were 5.98,3.94,2.05,2.13,1.39,1.24 μg/mL,and the LOD were 2.01,2.11,1.18,1.56,1.15,0.01 μg/mL,respectively.RSDs of precision tests were all less than 2.0％,stability and repetitive tests only ethyl acetate was detected,RSD＜2.0％;the recoveries were 93.59％-99.02％ (RSD=2.62％,n=6),92.42％-98.40％ (RSD=2.43％,n=6),94.81％-104.64％ (RSD=3.47 ％,n=6),94.56％-106.73％ (RSD=4.21％,n=6),97.04％-106.33％(RSD=3.50％,n=6)and 98.40％-107.97％ (RSD=3.37％,n=6).CONCLUSIONS:The method is specific,rapid,simple and accurate,and can be used for simultaneous determination of 6 residual organic solvents in Xingnaojing injection.

Simultaneous Determination of 6 Residual Organic Solvents in Xingnaojing Injection by Headspace GC / 中国药房

OBJECTIVE:To establish the method for simultaneous determination of 6 residual organic solvents in Xingnaojing injection,such as methanol,ethanol,isopropanol,n-butanol,ethyl acetate and acetonitrile.METHODS:Headspace GC method was adopted.The determination was performed on DB-624 capillary column by temperature programming with the injector temperature of 200 ℃;flame ionization detector was adopted with the temperature of 250 ℃;carrier gas was nitrogen with flow rate of 25 mL/min and split ratio of 35 ∶ 1;headspace sampling size was 1 mL,and heating temperature of headspace sampling was 80 ℃;equilibrium time was 15 min.RESULTS:The linear ranges of methanol,ethanol,isopropanol,n-butanol,ethyl acetate and acetonitrile were 15.00-240.00 μg/mL (r =0.999 9),25.00-400.00 μg/mL (r =0.999 9),25.00-400.00 μg/mL (r =0.999 9),25.00-399.99 μg/mL(r=0.999 9),25.00-399.99 μg/mL(r=0.999 8) and 5.00-80.00 μg/mL(r=0.999 9).The LOQ were 5.98,3.94,2.05,2.13,1.39,1.24 μg/mL,and the LOD were 2.01,2.11,1.18,1.56,1.15,0.01 μg/mL,respectively.RSDs of precision tests were all less than 2.0％,stability and repetitive tests only ethyl acetate was detected,RSD＜2.0％;the recoveries were 93.59％-99.02％ (RSD=2.62％,n=6),92.42％-98.40％ (RSD=2.43％,n=6),94.81％-104.64％ (RSD=3.47 ％,n=6),94.56％-106.73％ (RSD=4.21％,n=6),97.04％-106.33％(RSD=3.50％,n=6)and 98.40％-107.97％ (RSD=3.37％,n=6).CONCLUSIONS:The method is specific,rapid,simple and accurate,and can be used for simultaneous determination of 6 residual organic solvents in Xingnaojing injection.

Comparison of Efficacy of Salmeterol-flutiacasone Alone versus Fluticasone Propionate Combined with Mon-telukast Sodium in the Treatment of Children with Moderate Persistent Asthma / 中国药房

OBJECTIVE:To compare the clinical efficacy and safety of salmeterol-fluticasone alone versus fluticasone propio-nate combined with montelukast sodium in the treatment of children with moderate persistent asthma. METHODS:275 children with moderate persistent asthma were randomly divided into inhaled corticosteroids(ICS)+ long acting β2 receptor agonist(LABA) group (139 cases) and ICS+ leukotriene receptor antagonists (LTRA) group (136 cases). ICS+LABA group was given Salmeter-ol-fluticasone powder inhalant,1 inhalation,bid;ICS+LTRA was given Fluticasone propionate inhalation aerosol,bid+Montelu-kast sodium chewable tablet 5 mg,before bedtime,qd. Both groups received more than 12 weeks of treatment. Body indexes of 2 groups were observed after treatment,such as asthma control degree,asthma score,the percentage of peak expiratory flow(PEF) in estimated value,aberration rate of PEF,the levels of eosinophi cationic protein(ECP)and leukotrienes E4(LTE4)in peripheral blood,the times of taking short acting β2 receptor agonist (SABA) during treatment,asymptomatic days,compliance,the inci-dence of ADR,etc. RESULTS:Total effective rate of ICS+LABA group(86.33%)after 4 weeks treatment was higher than that of ICS+LTRA group(58.09%),with statistical significance(P0.05). The day and night asthma score of 2 groups decreased significantly and the percentage of PEF in estimated value increased significantly after treatment,there was statisti-cal significance compared to before treatment(P0.05). The levels of ECP and LTE4 decreased significantly in 2 groups after treatment,there was statistical signif-icance compared to before treatment(P0.05). CONCLUSIONS:Salmeterol-fluticasone alone versus fluticasone propionate combined with montelukast sodi-um in the treatment of children with moderate persistent asth-ma both have good therapeutic efficacy and safety,while thelatter one has stronger inhibition effect on ECP and LTE4,and to control asthma symptoms more rapidly.

Effect of tet methylcytosine dioxygenase 2 on the regulation of transforming growth factor-β1 expression in mesangial cells induced by high glucose / 中华肾脏病杂志

Objective To investigate the role of tet methylcytosine dioxygenase 2 (TET2) in the regulation of transforming growth factor-β1 (TGF-β1) expression in human glomerular mesangial cells induced by high glucose.Methods Cultured human glomerular mesangial cells were divided into normal control group (5.5 mmol/L glucose) and high glucose group (30.0 mmol/L glucose) which was cultured for 12 h to 72 h.The gene expression of TET2 in mesangial cells were inhibited by small molecule chemical called SC1,and which were divided into high glucose group (30.0 mmol/L glucose+ DMEM),DMSO group (30.0 mmol/L glucose+0.1％DMSO) and SC1 group (30.0 mmol/L glucose+3 μmol/L SC1).The mRNA and protein expression of TGF-β1,TET1 to 3 and α-smooth muscle actin (α-SMA) was detected by quantitative real-time PCR and Western blotting.Methylation of CpG islands in the regulation region of TGF-β1 was detected by bisulfite sequencing PCR (BSP).The activity of mesangial cell proliferation was assessed by colorimetry of thiazolyl blue (MTT).Results Compared with normal control group,the mRNA and protein expression of TET2 in mesangial cells induced by high glucose was increased significantly in a time-dependent manner (all P ＜ 0.05),but the expression of TET1 and TET3 was not affected.Meanwhile methylation rate of 4 CG sites from 24 h to 72 h were decreased in the first exon of TGF-β1 (P ＜ 0.01),but not in the promoter.Compared with high glucose group,when the expression of TET2 was inhibited by SC1,the methylation rate of TGF-β1 was recovered evidently (P ＜ 0.05),the mRNA and protein expression of TGF-β1 and α-SMA was suppressed,and the proliferation of mesangial cells was decreased (all P ＜ 0.05).Conclusions Demethylation of the CpG island mediated by TET2 may play an important role in the expression of TGF-β1 and mesangial cell phenotype transformation induced by high glucose.

Analysis of Problems with cultivating medical morality for medical students and relevant reasons / 中国医学伦理学

The research carried out a questionnaire regarding education of medical morality at four medical colleges in Fujian province, it found some problems existed, for example, the education of medical morality is ful-filled without knowing medical classical works, being short of systematicness, having no reviewing and encouraging mechanism. The article discusses the cause of the problem. ,with the reality of theInternet +, the characteris-tics of the medical students′education pattern and the characteristics of the medical ethics education.