Currently,human health due to corona virus disease 2019(COVID-19)pandemic has been seriously threatened.The coronavirus severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)spike(S)protein plays a crucial role in virus transmission and several S-based therapeutic approaches have been approved for the treatment of COVID-19.However,the efficacy is compromised by the SARS-CoV-2 evolvement and mutation.Here we report the SARS-CoV-2 S protein receptor-binding domain(RBD)inhibitor licorice-saponin A3(A3)could widely inhibit RBD of SARS-CoV-2 variants,including Beta,Delta,and Omicron BA.1,XBB and BQ1.1.Furthermore,A3 could potently inhibit SARS-CoV-2 Omicron virus in Vero E6 cells,with EC50 of 1.016 pM.The mechanism was related to binding with Y453 of RBD deter-mined by hydrogen-deuterium exchange mass spectrometry(HDX-MS)analysis combined with quan-tum mechanics/molecular mechanics(QM/MM)simulations.Interestingly,phosphoproteomics analysis and multi fluorescent immunohistochemistry(mIHC)respectively indicated that A3 also inhibits host inflammation by directly modulating the JNK and p38 mitogen-activated protein kinase(MAPK)path-ways and rebalancing the corresponding immune dysregulation.This work supports A3 as a promising broad-spectrum small molecule drug candidate for COVID-19.

Objective:To establish a rapid pipeline for whole genome sequencing of Chikungunya virus (CHIKV) by combining imbricated multiplex-PCR amplification and Illumina high-throughput sequencing platform.Methods:The primary reference sequences of CHIKV were downloaded from the National Center for Biotechnology Information (NCBI) database, covering all genotypes of CHIKV. After multiple alignments using the Mafft software and phylogenetic analysis, the 20 CHIKV references were selected for primer design. The Primal Scheme tool and Geneious Prime software were used to design, evaluate and optimize the primer panel. Finally, seven CHIKV-positive samples were involved in the validation of the primer panel.Results:All the amplicons of the designed panel were generated successfully. The consensuses generated from the mapping results could cover 100.00% of the coding region of the CHIKV genome when the Ct-value of the sample was less than 33, as the percentage would decrease to 99.38% when the Ct-value reached 35. The mapping percentage could be increased by 5.70%-25.43% when using the stepwise correction mapping strategy.Conclusion:The multiplex-PCR amplification method for CHIKV whole genome sequencing is relatively simple and convenient, which only requires two tubes of PCR amplification and performs well on CHIKV-positive clinical samples with different concentration levels of virus.

Objective:To establish a rapid pipeline for whole genome sequencing of Chikungunya virus (CHIKV) by combining imbricated multiplex-PCR amplification and Illumina high-throughput sequencing platform.Methods:The primary reference sequences of CHIKV were downloaded from the National Center for Biotechnology Information (NCBI) database, covering all genotypes of CHIKV. After multiple alignments using the Mafft software and phylogenetic analysis, the 20 CHIKV references were selected for primer design. The Primal Scheme tool and Geneious Prime software were used to design, evaluate and optimize the primer panel. Finally, seven CHIKV-positive samples were involved in the validation of the primer panel.Results:All the amplicons of the designed panel were generated successfully. The consensuses generated from the mapping results could cover 100.00% of the coding region of the CHIKV genome when the Ct-value of the sample was less than 33, as the percentage would decrease to 99.38% when the Ct-value reached 35. The mapping percentage could be increased by 5.70%-25.43% when using the stepwise correction mapping strategy.Conclusion:The multiplex-PCR amplification method for CHIKV whole genome sequencing is relatively simple and convenient, which only requires two tubes of PCR amplification and performs well on CHIKV-positive clinical samples with different concentration levels of virus.

Objective To evaluate the efficacy of superselective embolization of the supplying arteries of uterine fibroids under the guidance of 3D angiography in treating uterine fibroids.Methods The clinical data of 14 patients with uterine fibroids,who received interventional embolization therapy at the Third Affiliated Hospital of Zhengzhou University of China,were retrospectively analyzed.Of the 14 patients,5 received microcatheter superselective embolization of the supplying arteries of uterine fibroids after the origin and path of the supplying arteries were clarified by 3D uterine artery angiography(group A),and 9 received embolization of the bilateral uterine arteries up to their main trunks because the fibroids had complex blood supply(group B).The sums of the preoperative and postoperative one-year maximum diameter of uterine fibroids,and the reduction rate of fibroid were statistically analyzed.Results The mean sum of the preoperative and postoperative one-year maximum diameter of uterine fibroids in group A were(85.00±43.35)mm and(35.20±25.96)mm respectively,and the difference was statistically significant(P=0.006),which in group B were(65.00±12.68)mm and(49.44±24.83)mm respectively,and the difference was not statistically significant(P=0.052).There was no statistically significant differences in the mean sum of the preoperative maximum diameter of uterine fibroids and in the postoperative one-year maximum diameter of uterine fibroids between the two groups(P=0.366 and P=0.331).The median reduction rate of uterine fibroids in group A and group B was 62％and 25％respectively,and the difference was statistically significant(P=0.031).Conclusion For the treatment of uterine fibroids,3D angiography-guided superselective embolization of the supplying arteries of uterine fibroids is superior to the embolization of the bilateral uterine arteries in clinical curative efficacy.(J Intervent Radiol,2024,33:533-536)

Cryoablation (CRA) and microwave ablation (MWA) are two main local treatments for hepatocellular carcinoma (HCC). However, which one is more curative and suitable for combining with immunotherapy is still controversial. Herein, CRA induced higher tumoral PD-L1 expression and more T cells infiltration, but less PD-L1^highCD11b⁺ myeloid cells infiltration than MWA in HCC. Furthermore, CRA had better curative effect than MWA for anti-PD-L1 combination therapy in mouse models. Mechanistically, anti-PD-L1 antibody facilitated infiltration of CD8⁺ T cells by enhancing the secretion of CXCL9 from cDC1 cells after CRA therapy. On the other hand, anti-PD-L1 antibody promoted the infiltration of NK cells to eliminate PD-L1^highCD11b⁺ myeloid cells by antibody-dependent cell-mediated cytotoxicity (ADCC) effect after CRA therapy. Both aspects relieved the immunosuppressive microenvironment after CRA therapy. Notably, the wild-type PD-L1 Avelumab (Bavencio), compared to the mutant PD-L1 atezolizumab (Tecentriq), was better at inducing the ADCC effect to target PD-L1^highCD11b⁺ myeloid cells. Collectively, our study uncovered the novel insights that CRA showed superior curative effect than MWA in combining with anti-PD-L1 antibody by strengthening CTL/NK cell immune responses, which provided a strong rationale for combining CRA and PD-L1 blockade in the clinical treatment for HCC.

OBJECTIVE To explore the effects of naringin on the proliferation and differentiation of osteoblasts in T cell - osteoblast co -culture system under simulated microgravity . METHODS This experiment was setting normal gravity control group ， normal gravity naringin group ，simulated microgravity control group and simulated microgravity naringin group .In the control group，T cells were co -cultured with osteoblasts under normal gravity or simulated microgravity conditions . In the naringin group ， 1×10－5 mol/L naringin solution was added additionally on the basis of control group .The morphology of osteoblasts was observed under the microscope . The proliferation rate of osteoblasts was detected by CCK -8 method. The alkaline phosphatase （ALP）activity was detected by ALP kit . mRNA relative expressions of Runt related transcription factor 2（Runx2）and interleukin -6（IL-6）were detected by real -time fluorescence quantitative polymerase chain reaction . The relative expression of Runx 2 and IL -6 protein were detected by Western blot . RESULTS Compared with normal gravity control group ，the density of osteoblasts in the simulated microgravity control group decreased ，only a small number of cells were aggregated ，and the cell morphology was mostly round ； compared with the simulated microgravity control group ，the osteoblasts in the simulated microgravity naringin group formed spindle or polygonal ，plump in shape ，and clustered in groups of fish . Compared with normal gravity control group ，the proliferation rate of osteoblasts ，ALP activity ，relative expressions of Runx 2 mRNA and protein in the normal gravity naringin group were significantly increased ，while the relative expressions of IL -6 mRNA and protein were significantly decreased （P＜ 0.05）；the proliferation rate of osteoblasts ，ALP activity ，relative expressions of Runx 2 mRNA and protein in the simulated microgravity control group were significantly decreased ，while relative expressions of IL -6 mRNA and protein were significantly increased（P＜0.05）. Compared with the simulated microgravity control group ，the proliferation rate of osteoblasts ，ALP activity ， relative expressions of Runx 2 mRNA and protein in the simulated microgravity naringin group were significantly increased ，while relative expressions of IL -6 mRNA and protein were significantly decreased （P＜0.05）. CONCLUSIONS In the T cell -osteoblast co-culture system ，simulated microgravity can inhibit the proliferation and differentiation of osteoblasts ；naringin can improve the proliferation and differentiation ability of osteoblasts under this condition， and its mechanism of action is related to the decrease of IL -6 level and the increase of Runx 2 level.

It is an urgent demand worldwide to control the coronavirus disease 2019 (COVID-19) pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus. The 3-chymotrypsin-like protease (3CL^pro) and papain-like protease (PL^pro) are key targets to discover SARS-CoV-2 inhibitors. After screening 12 Chinese herbal medicines and 125 compounds from licorice, we found that a popular natural product schaftoside inhibited 3CL^pro and PL^pro with IC₅₀ values of 1.73 ± 0.22 and 3.91 ± 0.19 μmol/L, respectively, and inhibited SARS-CoV-2 virus in Vero E6 cells with EC₅₀ of 11.83 ± 3.23 μmol/L. Hydrogen-deuterium exchange mass spectrometry analysis, quantum mechanics/molecular mechanics calculations, together with site-directed mutagenesis indicated the antiviral activities of schaftoside were related with non-covalent interactions with H41, G143 and R188 of 3CL^pro, and K157, E167 and A246 of PL^pro. Moreover, proteomics analysis and cytokine assay revealed that schaftoside also regulated immune response and inflammation of the host cells. The anti-inflammatory activities of schaftoside were confirmed on lipopolysaccharide-induced acute lung injury mice. Schaftoside showed good safety and pharmacokinetic property, and could be a promising drug candidate for the prevention and treatment of COVID-19.

Background::Ubiquitination plays an essential role in many biological processes, including viral infection, and can be reversed by deubiquitinating enzymes (DUBs). Although some studies discovered that DUBs inhibit or enhance viral infection by various mechanisms, there is lack of information on the role of DUBs in virus regulation, which needs to be further investigated.Methods::Immunoblotting, real-time polymerase chain reaction, in vivo/ in vitro deubiquitination, protein immunoprecipitation, immunofluorescence, and co-localization biological techniques were employed to examine the effect of ubiquitin-specific protease 3 (USP3) on APOBEC3G (A3G) stability and human immunodeficiency virus (HIV) replication. To analyse the relationship between USP3 and HIV disease progression, we recruited 20 HIV-infected patients to detect the levels of USP3 and A3G in peripheral blood and analysed their correlation with CD4 + T-cell counts. Correlation was estimated by Pearson correlation coefficients (for parametric data). Results::The results demonstrated that USP3 specifically inhibits HIV-1 replication in an A3G-dependent manner. Further investigation found that USP3 stabilized 90% to 95% of A3G expression by deubiquitinating Vif-mediated polyubiquitination and blocking its degradation in an enzyme-dependent manner. It also enhances the A3G messenger RNA (mRNA) level by binding to A3G mRNA and stabilizing it in an enzyme-independent manner. Moreover, USP3 expression was positively correlated with A3G expression ( r= 0.5110) and CD4 + T-cell counts ( r= 0.5083) in HIV-1-infected patients. Conclusions::USP3 restricts HIV-1 viral infections by increasing the expression of the antiviral factor A3G. Therefore, USP3 may be an important target for drug development and serve as a novel therapeutic strategy against viral infections.

Image-guided radiation therapy using magnetic resonance imaging (MRI) is a new technology that has been widely studied and developed in recent years. The technology combines the advantages of MRI imaging, and can offer online real-time tracking of tumor and adjacent organs at risk, as well as real-time optimization of radiotherapy plan. In order to provide a comprehensive understanding of this technology, and to grasp the international development and trends in this field, this paper reviews and summarizes related researches, so as to make the researchers and clinical personnel in this field to understand recent status of this technology, and carry out corresponding researches. This paper summarizes the advantages of MRI and the research progress of MRI linear accelerator (MR-Linac), online guidance, adaptive optimization, and dosimetry-related research. Possible development direction of these technologies in the future is also discussed. It is expected that this review can provide a certain reference value for clinician and related researchers to understand the research progress in the field.
Magnetic Resonance Imaging ; Particle Accelerators ; Radiometry ; Radiotherapy Planning, Computer-Assisted ; Radiotherapy, Image-Guided

Objective:To evaluate the gastric emptying of orally administered enzyme-hydrolyzed rice flour solution before surgery in the patients undergoing laparoscopic cholecystectomy and effect on insulin resistance.Methods:One hundred patients, of American Society of Anesthesiologists physical status Ⅰ or Ⅱ, aged 18-64 yr, with body mass index of 19-30 kg/m 2, scheduled for elective laparoscopic cholecystectomy under general anesthesia, were divided into 2 groups ( n=50 each) using a random number table method: water group (group C) and enzyme-hydrolyzed rice flour group (group M). Routine fasting and water deprivation were executed at 1 day before operation in two groups, and 300 ml water in group C or 300 ml enzyme-hydrolyzed rice flour solution in group M were taken orally at 2-3 h before induction on the day of surgery.Bedside antrum ultrasonography was used to calculate the gastric volume (GV) before oral administration (V 0), immediately after oral administration (V 1), and before induction (V 2), and then the ΔGV (GV 1-GV 0) was calculated.Fasting plasma glucose and insulin CONCENTRATIONS were measured on admission to hospital (T 1) and on an empty stomach on 1st morning after surgery (T 2), and then the homeostasis model assessment of insulin resistance (HOMA-IR) was calculated according to HOMA steady-state model formula.Visual analog scale (VAS) scores for subjective comfort (thirst, hunger, fatigue and anxiety) and grip strength were assessed before anesthesia (T 3) and before leaving PACU (T 4). Reflux and aspiration during induction, nausea and vomiting within 24 h after surgery, and anal exhaust time after surgery were recorded. Results:There was no significant difference in GV at V 0, V 1 and V 2 between the two groups ( P>0.05). Compared with the baseline at V 0, no significant was found in the GV at V 2 in both groups ( P>0.05). The fasting plasma glucose and insulin concentrations and HOMA-IR were significantly increased at T 2 than at T 1 in both groups ( P<0.05 or 0.01). Compared with group C, the fasting plasma glucose and insulin concentrations and HOMA-IR were significantly decreased at T 2, VAS scores for hunger, fatigue and anxiety were decreased at T 3, 4, grip strength was increased at T 3, 4, the postoperative anal exhaust time was shortened, and the incidence of nausea was reduced in group M ( P<0.05). No reflux and aspiration happened during induction in either group. Conclusion:The gastric emptying of 300 ml enzyme-hydrolyzed rice flour solution orally administered at 2 h before surgery is normal in the patients undergoing laparoscopic cholecystectomy, which does not increase the risk of reflux and aspiration during anesthesia induction, reduces postoperative insulin resistance, and increases patient′s subjective comfort, and enhances the postoperative recovery of intestinal function.