A tandem-repeat dimeric RBD protein-based COVID-19 vaccine ZF2001 protects mice and nonhuman primates

A safe, efficacious and deployable vaccine is urgently needed to control COVID-19 pandemic. We report here the preclinical development of a COVID-19 vaccine candidate, ZF2001, which contains tandem-repeat dimeric receptor-binding domain (RBD) protein with alum-based adjuvant. We assessed vaccine immunogenicity and efficacy in both mice and non-human primates (NHPs). ZF2001 induced high levels of RBD-binding and SARS-CoV-2 neutralizing antibody in both mice and NHPs, and also elicited balanced TH1/TH2 cellular responses in NHPs. Two doses of ZF2001 protected Ad-hACE2-transduced mice against SARS-CoV-2 infection, as detected by reduced viral RNA and relieved lung injuries. In NHPs, vaccination of either 25 g or 50 g ZF2001 prevented infection with SARS-CoV-2 in lung, trachea and bronchi, with milder lung lesions. No evidence of disease enhancement is observed in both models. ZF2001 is being evaluated in the ongoing international multi-center Phase 3 trials (NCT04646590) and has been approved for emergency use in Uzbekistan.

Safety and immunogenicity of a recombinant tandem-repeat dimeric RBD protein vaccine against COVID-19 in adults: pooled analysis of two randomized, double-blind, placebo-controlled, phase 1 and 2 trials

BackgroundA safe and effective coronavirus disease 2019 (COVID-19) vaccine is urgently needed to control the ongoing pandemic. Although progress has been made recently with several candidates reporting positive efficacy results, COVID-19 vaccines developed so far cannot meet the global vaccine demand. We developed a protein subunit vaccine against COVID-19, using dimeric form of receptor-binding domain (RBD) as the antigen. We aimed to assess the safety and immunogenicity of this vaccine in humans and determine the appropriate dose and schedule for an efficacy study. MethodsWe did two randomized, double-blind, placebo-controlled, phase 1 and 2 trials for an RBD-based protein subunit vaccine, ZF2001. In phase 1 study, 50 healthy adults aged 18-59 years were enrolled and randomly allocated to three groups to receive three doses of vaccine (25 g or 50 g RBD-dimer, with adjuvant) or placebo (adjuvant-only) intramuscularly, 30 days apart. In phase 2 study, 900 healthy adults aged 18-59 years were enrolled and randomly allocated to six groups to receive vaccine (25 g or 50 g RBD-dimer, with adjuvant) or placebo (adjuvant-only) intramuscularly, with the former 3 groups given two doses and the latter 3 groups given three doses, 30 days apart. For phase 1 trial, the primary outcome was safety, as measured by the occurrence of adverse events and serious adverse events. The secondary outcome was immunogenicity as measured by the seroconversion rate and magnitude of antigen-binding antibodies, neutralizing antibodies and T-cell cytokine production. For phase 2 trial, the primary outcome included both safety and immunogenicity. These trials are registered with ClinicaTrials.gov, NCT04445194 and NCT04466085. FindingsBetween June 22 and September 15, 2020, 50 participants were enrolled to the phase 1 study (mean age 32.6 years) and 900 participants were enrolled to phase 2 study (mean age 43.5 years), to receive vaccine or placebo with a two-dose or three-dose schedule. For both trials, local and systemic adverse reactions were absent or mild in most participants. There were no serious adverse events related to vaccine in either trial. After three doses, neutralizing antibodies were detected in all participants receiving either 25 g or 50 g dose of vaccine in phase 1 study, and in 97% (the 25 g group) and 93% (the 50 g group) of participants, respectively, in phase 2 study. The SARS-CoV-2-neutralizing geometric mean titres (GMTs) were 94.5 for the 25 g group and 117.8 for the 50 g group in phase 1, and 102.5 for the 25 g group and 69.1 for the 50 g group in phase 2, exceeding the level of a panel of COVID-19 convalescent samples (GMT, 51). Vaccine induced balanced TH1 and TH2 responses. The 50 g group did not show enhanced immunogenicity compared with the 25 g group. InterpretationThe protein subunit vaccine ZF2001 is well-tolerated and immunogenic. The safety and immunogenicity data from phase 1 and 2 trials for ZF2001 support the use of 25 g vaccine dose with three-dose schedule to an ongoing phase 3 large-scale evaluation for safety and efficacy. FundingNational Program on Key Research Project of China, National Science and Technology Major Projects of Drug Discovery, Strategic Priority Research Program of the Chinese Academy of Sciences, and Anhui Zhifei Longcom Biopharmaceutical.

Correlation of body weight and its change with albuminuria in physical examination population / 中华肾脏病杂志

Objective:To study the effect of baseline weight and its change on new-onset albuminuria or increased urine albumin/creatinine ratio (ACR) in the physical examination population.Methods:The subjects of this study were those who completed two or more physical examinations at the Physical Examination Center of Sichuan Provincial People's Hospital from September 1, 2013 to September 1, 2018. The general information and laboratory examination results at the first and last physical examinations were collected. According to body mass index (BMI), they were divided into normal BMI group and overweight/obese group. The differences in general clinical data and laboratory test results between the two groups were compared. The primary endpoint events were new-onset albuminuria or urine ACR increase≥30%. Stepwise multiple linear regression method was used to analyze the influencing factors for ACR increase, and Cox proportional hazard model method was used to analyze the impact of baseline weight and its change on new-onset albuminuria or ACR increase ≥30%.Results:A total of 1 761 physical examination subjects were included in this study. The follow-up time was (16.54±7.87) months. There were 59 patients with new-onset albuminuria, 30 patients with ACR increase≥30%, and 35 patients with albuminuria reversal. Multiple linear regression analysis showed that BMI was an independent influencing factor for ACR ( β=0.127, P<0.001). Cox regression analysis showed that the older age ( HR=1.041, 95% CI 1.018-1.064, P<0.001), hypertension ( HR=2.035, 95% CI 1.278-3.242, P=0.003), diabetes ( HR=2.081, 95% CI 1.310-3.305, P=0.002) and hyperuricemia ( HR=1.700, 95% CI 1.084-2.668, P=0.021) were independent influencing factors for new-onset albuminuria or ACR increase≥30%, while BMI ( HR=1.053, 95% CI 0.975-1.137, P=0.191) and weight change rate ( HR=1.030, 95% CI 0.972-1.092, P=0.322) were not independent influencing factors for endpoint events. Subgroup analysis indicated that overweight/obesity had interactions with age, hypertension, diabetes, and hyperuricemia, respectively ( P for interaction<0.05), and the effects of overweight/obesity on the pre-set primary endpoint events in each subgroup were basically consistent. There were interactions between weight gain and hypertension and diabetes ( P for interaction<0.05). Weight gain increased the risk of the primary endpoint events of women ( HR=3.355, 95% CI 1.164-9.670, P=0.025), and the effects of overweight/obesity on the pre-set primary endpoint events of each subcomponent were basically the same (all P>0.05). The incidence of albuminuria reversal in the group with obvious weight loss was slightly higher than that in the group with obvious weight gain, but the difference was not statistically significant ( P>0.05), which might be related to the small weight loss range (-6.08%±3.51%). Conclusions:Overweight or obesity may increase the risk of albuminuria, and people with diabetes, hypertension, and hyperuricemia may be more likely to occur. Mild weight loss is not enough to reverse albuminuria.

Peri-procedural large-dose lactated Ringer solution in prevention and alleviation of post-ERCP pancreatitis: a meta-analysis / 中华肝胆外科杂志

Objective:To study the effectiveness and safety of perioperative lactated Ringer solution (LRS) in prevention and alleviation of post-endoscopic retrograde cholangiopancreatography pancreatitis (PEP) under different fluid replacement regimens to arrive at the most precise quantitative fluid replacement regimen.Methods:Pubmed, Embase, Cochrane Library Database, Wanfang Database, CNKI, and VIP were searched from inception to February 2020. Randomized controlled trials on LRS in prevention and alleviation of PEP under different fluid replacement regimens were collected. The experiment group was actively rehydrated with LRS during the perioperative period, and the amount of rehydration was significantly higher than that of the control group. The control group was given standard dose of LRS or normal saline. Two researchers independently selected the articles based on predetermined inclusion and exclusion criteria, extracted the data, and evaluated the risk of bias. RevMan 5.3 software was used for statistical analysis.Results:Ten studies with 2 261 patients were included, with 1 140 patients in the experiment group, and 1 121 patients in the control group. Meta-analysis showed that when the experiment group was given LRS at a rate of 5.0 ml·kg -1·h -1 during the perioperative period for about 9 hours and compared with the control group, the incidence of PEP in the experiment group was significantly reduced ( OR=0.32, 95% CI: 0.21-0.48, P<0.05). The incidence of moderate to severe PEP was also significantly reduced ( OR=0.28, 95% CI: 0.09-0.84, P<0.05). There was no increase in the incidence of adverse reactions related to fluid replacement. However, there were no significant differences in the incidence of PEP, and in moderate to severe PEP between the two groups when LRS was given at a rate of 4.0~4.5 ml·kg -1·h -1 within 9 hours, and less than 4.0 ml·kg -1·h -1 within 24 hours of total rehydration time (all P>0.05). Conclusion:During the perioperative period, the current evidence showed that it is most reasonable to give the fluid replacement regimen with aggressive hydration using LRS at a rate of 5.0 ml·kg -1·h -1 in about 9 hours to prevent and alleviate PEP. This is recommended for clinical practice and is worthy of further future studies.

Efficacy and safety of endoscopic papillary large balloon dilation alone versus endoscopic sphincterotomy combined with large balloon dilation in treatment of large common bile duct stones: A Meta-analysis / 临床肝胆病杂志

ObjectiveTo systematically review the efficacy and safety of endoscopic papillary large balloon dilation (EPLBD) versus endoscopic sphincterotomy combined with large balloon dilation (ESBD) in the treatment of large common bile duct stones (≥10 mm). MethodsPubmed, Embase, Cochrane Library, CNKI, Wanfang Data, and VIP were searched for related articles published up to March 2020. Two reviewers independently performed article screening, data extraction, and quality assessment, and RevMan 5.3 software was used for statistical analysis. ResultsA total of 11 studies (6 randomized controlled trials and 5 non-randomized controlled trials) were included, with 1282 patients in total. The meta-analysis showed that in the 6 randomized controlled trials, there were no significant differences between the EPLBD group and the ESBD group in initial stone clearance rate (odds ratio ［OR］=0.71, 95% confidence interval ［CI］: 0.45-1.12, P=0.14), overall stone clearance rate (OR=1.39, 95%CI: 0.65-2.96, P=0.39), rate of use of mechanical lithotripsy (OR=1.19, 95%CI: 0.75-1.89, P=0.46), and incidence rate of early complications (OR=1.10, 95%CI: 0.60-2.03, P=075); in the 5 non-randomized controlled trials, there were no significant differences between the EPLBD group and the ESBD group in initial stone clearance rate (OR=0.64, 95%CI: 0.35-1.15, P=0.13), overall stone clearance rate (OR=0.46, 95%CI: 0.19-112, P=009), and incidence rate of early complications (OR=1.20, 95%CI: 0.65-2.21, P=0.56), but the EPLBD group had a significantly higher rate of use of mechanical lithotripsy than the ESBD group (OR=1.96, 95%CI: 1.26-3.05, P=0.003). ConclusionEPLBD and ESBD have similar efficacy and safety in the treatment of large common bile duct stones, while EPLBD may increase the risk of the use of mechanical lithotripsy. More high-quality randomized controlled trials are needed to confirm this conclusion.

PTEN-silenced bone marrow mesenchymal stem cell transplantation for acute myocardial infarction / 中国组织工程研究

BACKGROUND:Bone marrow mesenchymal stem cel transplantation for myocardial infarction becomes popularized in recent years, but transplanted cels cannot survive and proliferate under early inflammatory reaction or local ischemia/hypoxia microenvironment, eventualy hampering the therapeutic outcomes. OBJECTIVE:To investigate the therapeutic effect of PTEN-silenced bone marrow mesenchymal stem cels on acute myocardial infarction. METHODS:(1) Bone marrow mesenchymal stem cels from Sprague-Dawley rats were randomly assigned to receive no treatment, NCsiRNA transfection using Lipofectamin2000orPTEN siRNA transfection using Lipofectamin2000. Cel growth curves were described using MTT method to detect cel cycle using flow cytometry. (2) Thirty Sprague-Dawley rats were selected to prepare myocardial infarction models that were randomized into three groups (n=10 per group): blank control, negative control and RNAi group. Six hours after modeling, bone marrow mesenchymal stem cels transfected with nothing, NCsiRNA and PTEN siRNA were respectively injected into the infarcted center of the left ventricular anterior wal in these three rat groups. After 4 weeks, al rats were subjected to cardiac function detection using echocardiography, and the survival and proliferation of bone marrow mesenchymal stem cels in the rats were observed by fluorescence microscopy. RESULTS AND CONCLUSION:Compared with the other two groups, a significant increase in the absorbance values at different culture time, the proportion of cels in S+G2phase, and the number ofbone marrow mesenchymal stem cels in the myocardial tissue was found in the RNAi group (alP< 0.05). Additionaly, the left ventricular ejection fraction and left ventricular shortening fraction were significantly reduced in the RNAi group than the blank control and negative control groups at 4 weeks after cel transplantation (P< 0.05). Bothin vivoandin vitroexperimental findings showed that PTEN silencing could effectively improve cel survival and proliferation in the infarcted myocardium. Moreover, in thein vivoexperiment, an overt improvement in rat’s cardiac function was achieved.

Association of oxygen uptake efficiency in exercise test with exercise capacity in patients of obstructive sleep apnea hypopnea syndrome / 中华全科医师杂志

Objective To explore the characteristics of oxygen uptake efficiency (OUES) in patients with obstructive sleep apnea hypopnea syndrome (OSAHS) and analyze the relationship between OUE and severity of disease.Methods Pulmonary function test,polysomnogram and cardiopulmonary exercise testing were performed in 35 patients with OSAHS and 25 age-matched healthy volunteers.Their successive breathing respiratory exchange parameters were collected and analyzed.And t and x2 tests were used for 2 sample comparison.Correlation analysis was performed by Pearson correlation test.Results Significant differences in peak VO2 and peak VO2 ％ pred existed between OSAHS and normal control groups [(18±4) vs.(28 ±6) L/min,P＜0.01;(68±14) vs.(84±16) ％,P＜0.01].Compared with normal control group [(2.3 ±0.5) L · min-1 · lg-1 ; (36 ±4) ml/L; (36 ±4) ml/L],OUES,OUEP and OUE@AT of OSAHS group [(1.8 ± 0.4) L · min-1 · lg-1 ; (31 ± 5) ml/L; (30 ± 5) ml/L] were significantly lower (t =3.78-4.49,all P ＜0.01).And OUES,OUEP and OUE@AT in OSAHS patients were correlated (r =0.53-0.67,all P ＜0.01) positively with exercise tolerance (peak VO2％ pred) while negatively with apnea hypopnea index (AHI) (r=-0.67--0.54,all P ＜0.01).Conclusion The oxygen uptake efficiency of patients with OSAHS is significantly reduced compared to that of normal subjects.And it is correlated negatively with severity of disease.

Expression of ubiquitin editing enzyme A20 and its pathway in steatotic hepatocytes and monocytes / 中华消化杂志

Objective To investigate the changes of A20 expression stimulated by free fatty acids (FFA) and its pathway.Methods HepG2 cells and U937 cells were stimulated by 0.5 mmol/L mixed FFA.The expression of A20,phosphor-p65 and phosphor-IκBα of neclear factor (NF)-κB pathway and phosphor-c-Jun N-terminal kinase (JNK),JNK,phosphor-extracellular signal-regulated kinase (ERK),ERK,phosphor-p38 and p38 of mitogen-activated protein kinase (MAPK) pathway were detected by Western blotting.The level of interleukin (IL)-12p,IL-1β,tumor necrosis factor (TNF)-α,IL-6,IL-10 and IL-8 cytokines in the supernatant of cell culture were detected by flow cytometry.T-test was performed for statistical analysis.Results The level of A20 changed along with the stimulated time of FFA.NF-κB and MAPK pathways were activated after FFA stimulation.The secretion of IL-6 and IL-8 increased after HepG2 cells stimulated by FFA and both reached peak at 24 hour.Compared with control group,the difference in IL-8 was statistically significant ((423.8 ± 8.9) pg/mL vs (12.4 ± 4.5) pg/mL,t=41.28,P＜0.01).The difference in IL-6 was also statistically significant ((4 082±423.6) pg/mL vs (52.9±29.5) pg/mL,t=9.49,P＜0.01).After U937 cells were stimulated by FFA,the secretion of IL-8 increased compared with control group.And in a certain period of time the secretion was time dependence.The maximum secretion of 24 hours was (200.6±5.7) pg/mL vs (5.0±3.9) pg/mL,and the difference was statistically significant (t=28.16,P＜0.01).IL-10,IL-12p,IL-1β and TNF-α were detected.Both NF-κB pathway and MAPK pathway were detected.Conclusions The in vitro FFA mediated steatotic cell model could induce the expression change of A20 and secretion of inflammatory cytokines.NF-κB and MAPK pathways involved in the response to FFA in HepG2 cells and U937 cells.

Ventilatory efficiency and exercise capacity during exercise in patients with idiopathic pulmonary fibrosis / 中华全科医师杂志

Objective To explore the characteristics of ventilatory efficiency and exercise capacity during cardiopulmonary exercise testing in patients with idiopathic pulmonary fibrosis (IPF).Methods Pulmonary function test,arterial blood gas analysis and cardiopulmonary exercise testing were performed in 28 IPF patients (IPF group) from April 2012 to April 2013 and 28 healthy volunteers (control group).And the relevant parameters were measured and compared.Results No significant differences existed in age [(57.8 ±9.8) vs.(59.2 ±5.5) years],gender or body mass index (BMI) [(23.8 ±2.7) vs.(25.0 ± 2.8) kg/m2,P ＞ 0.05].The paramneters of pulmonary function test,such as forced vital capacity ％ predicted (74.8 ± 14.6 vs.101.8 ± 10.8),forced expiratory volume in 1 second ％ predicted (73.8 ± 14.6 vs.97.0 ± 10.1),maximum voluntary ventilation ％ predicted (77.5 ± 14.9 vs.95.4 ±24.5),total lung capacity ％ predicted (75.6 ± 12.4 vs.99.8 ± 5.4),residual volume ％ predicted (80.7 ± 15.4 vs.95.8 ± 11.3),diffusing capacity of lung for carbon monoxide ％ predicted (66.2 ± 13.7 vs.103.2 ± 17.3) in the IPF group,were significantly lower than those of the control group (P ＜ 0.01).The parameters of arterial blood gas analysis,such as PaO2 [(72.7 ± 7.3) vs.(92.6 ± 3.8) mmHg] and SaO2 (94.3 ± 2.1 vs.98.3 ± 0.7),were lower than those of the control group (P ＜ 0.01).Thus P(A-a) O2 in the IPF group was higher than that in the control group (33.3 ± 5.7 vs.17.8 ± 1.9,P ＜0.01).These results strongly suggested that IPF group had restrictive ventilatory dysfunction and impaired gas exchange.The IPF patients had higher VE/VCO2-slope (37.4 ± 5.3 vs.25.7 ± 2.5,P ＜ 0.01) and lowest VE/VCO2 (39.2 ±6.7 vs.30.6 ± 2.7,P ＜ 0.01) than the controls; VE/VCO2 and VD/VT during every period were significantly higher in the IPF group than those in the control group (P ＜ 0.01) ; during peak exercise,peakLoad％pred (70.4 ±±29.9 vs.104.8 ±29.7,P ＜0.01) and peakVO2％pred (68.7 ±29.8 vs.98.7 ±36.4,P =0.001) were significantly lower in the IPF group than those in the control group.In the IPF group,VE/VCO2@AT,VE/VCO2-slope and lowest VE/VCO2 had a negative correlation with DLCO％pred (r=-0.589,P ＜0.01; r=-0.481,P＜0.05; r=-0.527,P＜0.05).In the IPF group,VE/VCO2@AT,VE/VCO2-slope and lowest VE/VCO2 had a negative correlation with peakVO2％ pred (r =-0.548,P ＜ 0.05 ; r =-0.539,P ＜ 0.05 ; r =-0.564,P ＜ 0.05).So the exercise tolerance and ventilation efficiency of the IPF group decreased significantly.Conclusion Cardiopulmonary exercise testing reveals that the ventilation efficiency of IPF patients decreases significantly so as to seriously affect their exercise tolerance