BACKGROUND: Innovative coronavirus disease 2019 (COVID-19) vaccines, with elevated global manufacturing capacity, enhanced safety and efficacy, simplified dosing regimens, and distribution that is less cold chain-dependent, are still global imperatives for tackling the ongoing pandemic. A previous phase I trial indicated that the recombinant COVID-19 vaccine (V-01), which contains a fusion protein (IFN-PADRE-RBD-Fc dimer) as its antigen, is safe and well tolerated, capable of inducing rapid and robust immune responses, and warranted further testing in additional clinical trials. Herein, we aimed to assess the immunogenicity and safety of V-01, providing rationales of appropriate dose regimen for further efficacy study. METHODS: A randomized, double-blind, placebo-controlled phase II clinical trial was initiated at the Gaozhou Municipal Centre for Disease Control and Prevention (Guangdong, China) in March 2021. Both younger (n = 440; 18-59 years of age) and older (n = 440; ≥60 years of age) adult participants in this trial were sequentially recruited into two distinct groups: two-dose regimen group in which participants were randomized either to follow a 10 or 25 μg of V-01 or placebo given intramuscularly 21 days apart (allocation ratio, 3:3:1, n = 120, 120, 40 for each regimen, respectively), or one-dose regimen groups in which participants were randomized either to receive a single injection of 50 μg of V-01 or placebo (allocation ratio, 3:1, n = 120, 40, respectively). The primary immunogenicity endpoints were the geometric mean titers of neutralizing antibodies against live severe acute respiratory syndrome coronavirus 2, and specific binding antibodies to the receptor binding domain (RBD). The primary safety endpoint evaluation was the frequencies and percentages of overall adverse events (AEs) within 30 days after full immunization. RESULTS: V-01 provoked substantial immune responses in the two-dose group, achieving encouragingly high titers of neutralizing antibody and anti-RBD immunoglobulin, which peaked at day 35 (161.9 [95% confidence interval [CI]: 133.3-196.7] and 149.3 [95%CI: 123.9-179.9] in 10 and 25 μg V-01 group of younger adults, respectively; 111.6 [95%CI: 89.6-139.1] and 111.1 [95%CI: 89.2-138.4] in 10 and 25 μg V-01 group of older adults, respectively), and remained high at day 49 after a day-21 second dose; these levels significantly exceed those in convalescent serum from symptomatic COVID-19 patients (53.6, 95%CI: 31.3-91.7). Our preliminary data show that V-01 is safe and well tolerated, with reactogenicity predominantly being absent or mild in severity and only one vaccine-related grade 3 or worse AE being observed within 30 days. The older adult participants demonstrated a more favorable safety profile compared with those in the younger adult group: with AEs percentages of 19.2%, 25.8%, 17.5% in older adults vs. 34.2%, 23.3%, 26.7% in younger adults at the 10, 25 μg V-01 two-dose group, and 50 μg V-01 one-dose group, respectively. CONCLUSIONS: The vaccine candidate V-01 appears to be safe and immunogenic. The preliminary findings support the advancement of the two-dose, 10 μg V-01 regimen to a phase III trial for a large-scale population-based evaluation of safety and efficacy. TRIAL REGISTRATION http://www.chictr.org.cn/index.aspx (No. ChiCTR2100045107, http://www.chictr.org.cn/showproj.aspx?proj=124702).
Aged ; Antibodies, Viral ; COVID-19/therapy* ; COVID-19 Vaccines ; Double-Blind Method ; Humans ; Immunization, Passive ; Recombinant Fusion Proteins ; SARS-CoV-2

Objective:To establish a disease risk prediction model for the newborn screening system of inherited metabolic diseases by artificial intelligence technology.Methods:This was a retrospectively study. Newborn screening data ( n=5 907 547) from February 2010 to May 2019 from 31 hospitals in China and verified data ( n=3 028) from 34 hospitals of the same period were collected to establish the artificial intelligence model for the prediction of inherited metabolic diseases in neonates. The validity of the artificial intelligence disease risk prediction model was verified by 360 814 newborns ' screening data from January 2018 to September 2018 through a single-blind experiment. The effectiveness of the artificial intelligence disease risk prediction model was verified by comparing the detection rate of clinically confirmed cases, the positive rate of initial screening and the positive predictive value between the clinicians and the artificial intelligence prediction model of inherited metabolic diseases. Results:A total of 3 665 697 newborns ' screening data were collected including 3 019 cases ' positive data to establish the 16 artificial intelligence models for 32 inherited metabolic diseases. The single-blind experiment ( n=360 814) showed that 45 clinically diagnosed infants were detected by both artificial intelligence model and clinicians. A total of 2 684 cases were positive in tandem mass spectrometry screening and 1 694 cases were with high risk in artificial intelligence prediction model of inherited metabolic diseases, with the positive rates of tandem 0.74% (2 684/360 814)and 0.46% (1 694/360 814), respectively. Compared to clinicians, the positive rate of newborns was reduced by 36.89% (990/2 684) after the application of the artificial intelligence model, and the positive predictive values of clinicians and artificial intelligence prediction model of inherited metabolic diseases were 1.68% (45/2 684) and 2.66% (45/1 694) respectively. Conclusion:An accurate, fast, and the lower false positive rate auxiliary diagnosis system for neonatal inherited metabolic diseases by artificial intelligence technology has been established, which may have an important clinical value.

Objective To prepare F7 thermosensitive liposome and evaluate its physicochemical properties, then investigate its cytotoxicity against tumor cells in vitro. Methods The F7 thermosensitive liposome was prepared by the pH gradient active drug loading method using dipalmitoyl phosphatidylcholine myristoyl lyso-phosphocholine and 1, 2-distearoyl-sn-glycero-3-phosphoethanolamine-N-methoxy (polyethylene glycol)-2000 as membrane materials. The encapsulation efficiency and drug loading were determined for the F7 thermosensitive liposome by HPLC. The phase transition temperature of F7 thermosensitive liposome was investigated by differential scanning calorimetry;the liposome morphology was observed by atomic force microscopy;the drug release of liposome was examined by dialysis;and the particle size and zeta potential were measured through Malvern particle size analyzer. The cytotoxicity of F7 and F7 thermosensitive liposome was determined by the MTT method, and the freeze-drying process was optimized using the designexpert software. Results The encapsulation efficiency of F7 thermosensitive liposomes was (97.56±0.22) ％, and the drug loading ratio was (1.51±0.01) ％. The phase transition temperature of F7 thermosensitive liposome was 39.9℃, the zeta potential was (-15.10±0.85) mV, the particle size was (86.94±1.21) nm, and the poly disperse coefficient was 0.17±0.01. Compared with the F7 injection, the F7 thermosensitive liposomes showed a stronger, dose-dependent inhibitory effect on the growth of lung cancer H1299 and breast cancer MCF-7 cells. The freeze-dried powder of liposomes dissolved well with the encapsulation efficiency of 95％ and the particle size of approximately 130 nm. Conclusion The F7 thermosensitive liposome prepared by the pH gradient active drug loading method has high encapsulation efficiency and good stability. The preparation method is simple and feasible for further development of the F7 preparation.

Objective To study the effects of verbascoside on the proliferation and osteogenic differentiation of rat bone marrow stromal cells (BMSCs) and its mechanism.Methods BMSCs were obtained and identified from rat bone marrow by density gradient centrifugation.BMSCs were randomly divided into high,medium,low dose test groups and control group,and treated with 100,50,25,0 μmol · L-1verbascoside.The cell counting kit-8 (CCK-8) method was used to evaluate the BMSCs proliferation at 24,48,72 h after administration the medication.The alkaline phosphatase(ALP) activities were detected at 24,48,96 h after medication.The ALP staining was performed after 72 h of medication.Western blot and real time-polymerase chain reaction (PCR) were used to detect the osteogenic related proteins and genes expression.Results After 72 h,the proliferation of BMSCs in high dose test group was 1.22 ± 0.05,lower than that in control group,which was 1.50 ± 0.09 (P ＜ 0.05).The ALP expression levels in high,medium and low dose test groups were higher than control group at 24,48,96 h.After 72 h,ALP staining showed that the color of culture dishes in high,medium and low dose test groups were deeper than that in control group.Western blot showed that osteogenic related proteins of bone morphogenetic protein-2 (BMP2),Smad1,Smad5,Smad8,Runx2,Osterix in high,medium and low dose test groups were higher than those in control group (P ＜0.05),but there was no significant difference among those three test groups (P ＞ 0.05).Real time-PCR showed that the osteogenic related genes of BMP2,Smad1,Smad5,Smad8,Runx2,Osterix expression levels in high,medium and low dose test groups were higher than those in control group (P ＜ 0.05).However,there was no significant difference among three test groups(P ＞ 0.05).Conclusion The verbascoside at different concentrations has no obvious effect on BMSCs proliferation and promote osteogenic differentiation of BMSCs through BMP2-Smads -Runx2-Osterix pathway.

Aim To explore the effects of niacin on LDL-C uptake and metabolism in HepG2 cells,and to clarify the functions of niacin in lipid-lowering and slo-wing the atherosclerosis process,thus to provide a sci-entific basis for niacin as a lipid-lowering drug in clini-cal development.Methods Oil red O staining was used to observe HepG2 cells after lipid uptake.Enzy-matic method was used to determine the content of in-tracellular free cholesterol (FC)and total cholesterol (TC).The LDLR levels on the surface of cell mem-brane were detected by immunofluorescence flow cy-tometer.The mRNA and protein expressions of LDLR, SREBP2 and PCSK9 were analyzed by qPCR and Western blot.Results The results of oil red O staining showed that the rate of oil red O-positive cells and the number of red lipid droplets were significantly in-creased in niacin group than control group.Niacin sig-nificantly increased the levels of TC and FC in HepG2 cells(P <0.05 ).What’s more,niacin significantly upregulated the expression of LDLR and significantly downregulated the protein expression of PCSK9,while it had no effect on the expression of SREBP2.Conclu-sion Niacin accelerates LDL-C uptake probably via downregulating the expression of PCSK9 and reducing the degradation of LDLR protein in HepG2 cells.

AlM: To explore the expression and significance of cysteine- rich 61 ( CCN1/Cyr61 ) in oxygen - induced retinal neovascularization ( RNV) of mice and study the inhibition effect of CCN1 specific siRNA on RNV.
METHODS:Two hundred healthy C57BL/6J mice were chosen and randomly divided into control group, hyperxia group, hyperxia control group and CCN1 treated group, with 50 mice in each group. The hyperxia control group was treated with vector plasmids by intravitreal injection. The CCN1 treated group received CCN1 siRNA recombinant plasmids by intravitreal injection. Adenosine diphosphate-ase ( ADPase) stained retina flat-mounts was performed to assess the retinal vascular profiles, HE staining was applied to count the number of vascular endothelial cell nuclei breaking through the internal limiting membrane, protein and mRNA level expression of CCN1 were measured by immunohistochemistry, Western blot and RT-PCR.
RESULTS: There were large nonperfusion area and a large number of vascular endothelial cell nuclei breaking through the internal limiting membrane ( 25. 25 ± 1. 26;23. 12 ± 1. 16 ) in the hyperxia group and the hyperxia control group. Regions of nonperfusion and vascular endothelial cell nuclei (8. 47±1. 15) were decreased in the CCN1 treated group compared to the hyperxia group and the hyperxia control group. Compared with the control group, there were high protein and mRNA expression of CCN1 in the hyperxia group and the hyperxia control group. The expression of CCN1 protein and mRNA were decreased in the CCN1 treated group compared with the hyperxia group and hyperxia control group (all P<0. 05).
CONCLUSlON: The abnormal expression of CCN1 has close relation with RNV. The development of RNV can be markedly inhibited by RNA interference targeting CCN1, which, we believe, will provide new molecular targets and a rationale for clinical developing new strategy for ROP therapy.

Optimization of sensor array is a significant topic in the application of electronic nose (EN). Stepwise discriminant analysis and cluster analysis combining with screening of typical index were employed to optimize the original array in the classification of 100 samples from 10 kinds of traditional Chinese medicine based on alpha-FOX3000 EN. And the identification ability was evaluated by three algorithm including principle component analysis, Fisher discriminant analysis and random forest. The results showed that the identification ability of EN was improved since not only the effective information was maintained but also the redundant one was eliminated by the optimized array. The optimized method was eventually established, it was accurate and efficient. And the optimized array was built up, that is, S1, S2, S5, S6, S8, S12.
Algorithms ; Biosensing Techniques ; methods ; Cluster Analysis ; Discriminant Analysis ; Drugs, Chinese Herbal ; classification ; isolation & purification ; Electronic Nose ; Medicine, Chinese Traditional ; Principal Component Analysis ; Reproducibility of Results ; Smell