Coronavirus-related diseases pose a significant challenge to the global health system. Given the diversity of coronaviruses and the unpredictable nature of disease outbreaks, the traditional "one bug, one drug" paradigm struggles to address the growing number of emerging crises. Therefore, there is an urgent need for therapeutic agents with broad-spectrum anti-coronavirus activity. Here, we provide evidence that ATV006, an anti-SARS-CoV-2 nucleoside analog targeting RNA-dependent RNA polymerase (RdRp), has broad antiviral activity against human and animal coronaviruses. Using mouse hepatitis virus (MHV) and human coronavirus NL63 (HCoV-NL63) as a model, we show that ATV006 has potent prophylactic and therapeutic activity against murine coronavirus infection in vivo. Remarkably, ATV006 successfully inhibits viral replication in mice even when administered 96 h after infection. Due to its oral bioavailability and potency against multiple coronaviruses, ATV006 has the potential to become a useful antiviral agent against SARS-CoV-2 and other circulating and emerging coronaviruses in humans and animals.

Human pluripotent stem cells provide an inexhaustible model to study human embryogenesis in vitro. Recent studies have provided diverse models to generate human blastoids by self-organization of different pluripotent stem cells or somatic reprogramming intermediates. However, whether blastoids can be generated from other cell types or whether they can recapitulate postimplantation development in vitro is unknown. Here, we develop a strategy to generate human blastoids from heterogeneous intermediates with epiblast, trophectoderm, and primitive endoderm signatures of the primed-to-naïve conversion process, which resemble natural blastocysts in morphological architecture, composition of cell lineages, transcriptome, and lineage differentiation potential. In addition, these blastoids reflect many features of human peri-implantation and pregastrulation development when further cultured in an in vitro 3D culture system. In summary, our study provides an alternative strategy to generate human blastoids and offers insights into human early embryogenesis by modeling peri- and postimplantation development in vitro.
Humans ; Pluripotent Stem Cells/metabolism* ; Embryo, Mammalian/metabolism* ; Cell Differentiation ; Blastocyst ; Cell Lineage ; Embryonic Development

AIM: To observe the effect of acute normovolemic hemodilution (ANH) autologous blood transfusion on the EEG bispectral index and muscle relaxation in elderly patients undergoing orthopedic surgery to explore the influence of autologous blood transfusion containing anesthetic components on the quality and safety of postoperative anesthesia recovery. METHODS: Forty patients, aged 65-75, weighing 55-80 kg, ASA grade I-II, with an estimated intraoperative blood loss of more than 600 mL, were selected for elective orthopedic surgery. The patients were randomly divided into two groups (n=20): group A was given acute normovolemic hemodilution (ANH), and the target value of Hct was 28%-30% after induction of anesthesia; group B was the control group which was given routine fluid infusion during operation without ANH. Bispectral index (BIS), TOF values and plasma concentrations of propofol and cisatracurium were measured at the beginning of autotransfusion (T

OBJECTIVE To explore the mechanism of Aurora kinase A (Aurora-A) promoting cancer cell chemotherapy resistance in nasopharyngeal carcinoma. METHODS The expression of Aurora-A in nasopharyngeal carcinoma tissues and adjacent tissues were detected by Western bolt and Q-PCR. The highexpressing Aurora A cell line CNE2 was used to detected the cell apoptosis and the expression of key pathway marker protein after Aurora-A inhibitor VX680 and cisplatin treatment by using Flow cytometry and WB. RESULTS The expression of Aurora-A in nasopharyngeal carcinoma tissues was significantly higher than that in adjacent tissues. Comparing to normal nasopharyngeal cells NP69, Aurora-A was significantly highly expressed in all of nasopharyngeal carcinoma cells and was highest in CNE2. Inhibiton of Aurora-A increased the cell apoptosis and the expression of p-AKT, p21 and Cleaved-Caspase-3 after using cisplatin or the Aurora-A inhibitor VX680 treatment. CONCLUSION The results shown that Aurora-A confer chemoresistance to cisplatin treatment through p-AKT/p21/Cleaved-Caspase-3 pathway.

[Objective]To explore the effect of HSYA on pro/anti-inflammatory cytokine′s levels and mRNA expression in peripheral blood of mice.with sepsis.[Methods]Dividing NIH mice into four groups,as normal group,sham group,CLP group and HSYA group,24 mice in each group. The CLP sepsis mouse model was established. HSYA(120 mg/kg)were injected intravenously at 12 h before the operation ,and 0 h and 12 h following CLP ,and other groups were given normal saline. observed the animals behavior changes,measured the levels WBC,PLT,ALT,AST,BUN in serum,detected the levels and mRNA expression of IL-6, IL-10 ,TNF-α in peripheral blood. cultured of peripheral blood bacteria loads.[Results]24 h after surgery ,mice in CLP group appeared furring,feces residues on anus etc. compared to normal group,sham group and HSYA group,WBC,PLT,ALT,AST, BUN,levels and mRNA expression of IL-6,IL-10,TNF-αshowed significant increases,it was also found that bacterial load was significant increased in model group.[Conclusions]HSYA has a therapeutic effect in mice with sepsis ,can reduce bacteria into the blood,and inhibit inflammatory mediators which caused tissue damage.

Objective: To investigate the clinical features of imbalance between Th1 and Th22 cells and its association with disease progression in patients with liver cirrhosis, and to explore immune therapeutic strategies for targeted therapy for liver cirrhosis. Methods: In vitro peripheral blood mononucleated cells (PBMCs) were collected by centrifugation. CD3-BV500 and CD8-PerCP-Cy5.5 staining was performed for these cells. IFNγ-PE-Cy7, IL-17a-APC, IL-22-PE, or the corresponding isotype control was added, and then PBMCs were fixed with 1% polyoxymethylene after being washed once by permeabilization-wash buffer. Flowjo software was used for the analysis of T lymphocyte subsets and cytokines. Th1 (CD4+IFNγ⁺), Th17 (CD4+IL-17a⁺), Th22 (CD4+IL-22⁺), Tc1 (CD8+IFNγ⁺), Tc17 (CD8+IL-17a⁺), and Tc22 (CD8+IL-22⁺) subsets were defined and the secretions of interferon-γ (IFN-γ), interleukin-17a (IL-17a), and interleukin-22 (IL-22) were measured for all subsets. LX-2 cells were cultured in a serum-free medium and different concentrations of recombinant human IL-22 protein (25, 50, 100 ng/ml) were added; 24 hours later, the activation marker α-smooth muscle actin (α-SMA) was used to measure LX-2 activation. Fetal bovine serum with a volume fraction of 10% was used as a positive control. Enzyme-linked immunosorbent assay (chemiluminescence) was used to measure the concentrations of hyaluronic acid, type III precollagen, and type IV collagen in supernatant. A one-way analysis of variance, the non-parametric Mann-Whitney U test, and the non-parametric Kruskal-wallis H test were used for statistical analysis based on data type. Results: Compared with the health control group, the liver cirrhosis groups with various causes had significant increases in peripheral Tc1, Th17, and Th22 cells. The percentage of Th17 cells in the liver cirrhosis group was 1.64 times that in the control group (4.25%±2.45% vs 2.59%±1.36%, P < 0.05), and the mean percentage of Th22 cells in the liver cirrhosis group was 2.18 times that in the control group (4.17%±2.55% vs 1.31%±0.64%, P < 0.05). The percentages of Th17 (5.89%±3.44%) and Th22 cells (5.32%±3.67%) in the patients with alcoholic cirrhosis were 1.27 and 3.06 times those in the control group (P < 0.05). The patients with alcoholic cirrhosis had a significant increase in Th22 cells. The patients with different types of liver cirrhosis had a significant reduction in the ratio between anti-fibrotic and pro-fibrotic factors (Th1/Th22), which was positively correlated with the severity of liver cirrhosis and was a common immunological feature of liver cirrhosis with different causes. In addition, IL-22 activated hepatic stellate cells and promoted the production of collagen. Conclusion The imbalance between anti-fibrotic and pro-fibrotic factors (Th1/Th22) is a common feature of the progression of liver fibrosis with various causes and may contribute to the progression of liver fibrosis.

Objective To detect the expression level and procoagulant activity of tissue factor (TF)in the blood and urine in patients with type 2 diabetic nephropathy (DN),and investigate its related clinical significance.Methods A total of 120 DN patients were collected and divided into 3 groups according to urinary albumin and creatinine ratio:normal albuminuria group (＜3.4 mg/mmol) with 64 cases,microalbuminuria group ( ≥3.4 mg/nunol,and ＜ 34.1 mg/mmol) with 35 cases and macroalbuminuria group (≥34.1 mg/mnol) with 21 cases.Twenty healthy persons were enrolled as control group.The expression of monocytes TF in peripheral blood was detected by flow cytometric analyzer.The expression ofTF in the blood and urine was detected by ELLSA.The prcoagulant activity of TF was detected by one-stage coagulation.Results The expressions of monocytes TF in normal albuminuria group,microalbuminuria group and macroalbuminuria group [ (4.32 ± 1.25 )％,(5.01 ± 1.73)％,( 1 1.83 ± 2.14)％ ] were significantly higher than that in control group [(0.84 ± 0.36)％ ](P ＜0.01).TF procoagulant activity in normal albuminuria group,microalbuminuria group and macroalbuminuria group [ ( 64.3 ± 21.2),( 67.1 ± 17.3 ),( 113.8 ± 44.4)mU/L] were obviously higher than that in control grouP [ (23.8 ± 16.4) mU/L](P＜ 0.01 ).The expression of monocytes TF in normal albuminuria group and microalbuminuria group had no statistical significance (P＞0.05),while the expression in macroalbuminuria group had significant difference compared with that in normal albuminuria group and microalbuminuria group (P ＜ 0.01 ).The expression of TF in the blood and urine of DN patients was obviously higher than that of control group (P ＜ 0.01 ).The expression of TF in the blood was positively correlated with that in urine (r =0.684,P ＜ 0.01 ).Conclusions The expressions of TF in the blood and urine in DN patients increase and its procoagulant activity enhances.The expression level of TF has a closed relationship with the increase of albuminuria.

Objective To investigate the relation between Fas-mediated apoptosis and glucocorticoid treatment in myasthenia gravis (MG). Methods In 17 patients with MG, 6 patients received glucocorticoid treatment (glucocorticoid treatment group),and 11 patients were treated without glucocorticoid (nonglucocorticoid treatment group). Meanwhile, 13 healthy cases were selected as healthy control group. CD4,CD8 and Fas expressions in peripheral blood T lymphocyte were detected by flow cytometry in three groups and analyzed. Results The percentage of CD4-CD8+ cells in peripheral blood T lymphocyte in glucocorticoid treatment group was significantly higher than that in healthy control group[(36.75 ± 11.56)% vs. (26.31 ±9.00)%, P = 0.027], while the percentage of CD4-CD8- cells was significantly lower [(30.56 ± 9.72)% vs.(42.96 ± 11.54)%, P =0.018]. The percentage of CD4-CD8+ cells in peripheral blood T lymphocyte in glucocorticoid treatment group was significantly higher than that in non-glucocorticoid treatment group [(36.75 ± 11.56)% vs. (25.24 ±7.63)% ,P =0.019]. The percentages of Fas+ and CD8 +Fas+ cells in peripheral blood T lymphocyte in glucocorticoid treatment group were significantly higher than those in healthy control group[(46.10 ± 7.13)% vs. (31.22 ± 13.00)%, P=0.006; (62.86 ± 12.29)% vs. (45.59 ±11.50)%, P = 0.003]. The percentage of CD8+ Fas+ cells in peripheral blood T lymphocyte in glucocorticoid treatment group was significantly higher than that in non-glucocorticoid treatment group [(62.86 ± 12.29)%vs (50.84 ± 8.31 )%, P = 0.034]. Conclusions Glucocorticoid treatment may have influence on peripheral blood T lymphocyte subsets in patients with MG. Fas-mediated apoptosis may be involved in the mechanism of glucocorticoid treatment in MG.

Objective To study the efficacy of different anti-adhesion agents used in preventing tubal obstruction after recanalization.Methods Five hundred and eight patients with tubal obstruction were divided into 245 cases in control group,108 cases in chitosan group;113 cases in sodium hyaluronate group and 42 cases in lipiodol group.The patients in control group were injected with anti-inflammation agents after recanalizatian,while other groups were injected with chitosan,sodium hyaluronate or lipiodol at dose of 2-3 ml in every therapeutic group.The rate of location of tubal obstruction and tubal recanalization were recorded during operation.Then patients in every group were followed up on tubal patency after 3 months,and pregnancy rate after 12 months.Results Among 1016 fallopian tubes in 508 patients,there were 330 tubes occlusion at isthmus portion and 563 tubes occlusion at interstitial portion of fallopian tube.Thirtyseven fallopian tubes were ablated because of ectopic pregnancy,86 fallopian tubes were unobstructed.(1)The recanalization rate were 95.7% (179/187) in chitosan group,97.9% (191/195) in sodium hyaluronate group,98.7% (75/76) in lipiodol group and 97.7% (425/435) in control group,which did not show statistical difference (P＞0.05).(2) The rates of tubal patency after 3 months of 91.7% (99/108) in chitosan group and 88.5% (100/113) in sodium hyaluronate group were significantly higher than 71.4% (30/42) in lipiodol group and 74.3% (182/245) in control group (P ＜0.05).(3)The rates of intrauterine pregnancy after 12 months were 48.1% (52/108) in chitosan group and 41.6% (47/113) in sodium hyaluronate group,which were significantly higher than 23.8% (10/42) in lipiodol group and 24.1% (59/245) in control group (P ＜ 0.05).Conclusion Chitosan and sodium hyaluronate could be effective to prevent tubal obstruction after interventional recanalization and increase pregnancy rate.

Objective To study the expression of eytokines in acute hemorrhage patients with systenlic in-flammatory response syndrome(SIRS)and multiple organ dysfunction syndrome(MODS).Methods Platelet acti-vation markers CD62p and cytokines in acute hemorrhage patients with SIRS and MODS were detected by using flow cytometry,and compared with that of control group.Results The expression of CD62p and cytokines(TNF-α and IL-6)in acute hemorrhage patients,with or without SIRS and MODS wag significantly higher than that in control group(P<0.01),and higher in MODS group than in SIRS group,and in MODS death group than in MODS survival group(P<0.01).Conclusion The activation of platelet and the over-expression of cytokines participate in the on-set and development of acute hemorrhage,SIRS and MODS,and were related with the severity of disease and progno-sis.