OBJECTIVE To investigate the in vitro activity of Chansu injection against SARS-CoV-2 infection and determine whether bufalin is the primary active component responsible for its efficacy.METHODS The half-maximal effective concentration(EC50)of Chansu injection and bufalin against SARS-CoV-2 infection was determined using the CellTiter-Glo cell viability assay.qPCR was performed to assess the effects of Chansu injection and bufalin on mRNA levels of SARS-CoV-2 NP protein,inflammatory factors,and interferons in virus-infected cells.A cell-cell membrane fusion model was established,and the impact of the drugs on membrane fusion between HEK-293T and HEK-293T-ACE2 cells was evaluated using a luciferase reporter gene assay.RESULTS Chansu injection exhibited anti-SARS-CoV-2 virus infection activity,with an EC50 of 85.56 ng·mL-1.Chansu injec-tion significantly reduced the mRNA levels of viral NP protein(P<0.05),IFN-λ2/3(P<0.05),ISG-15 and RIG-I(P<0.000 1)in SARS-CoV-2 infected Calu-3 cells,and the mRNA levels of inflammatory factors IL-6 and TNF-α were significantly reduced(P<0.000 1).The EC50 of bufalin in inhibiting SARS-CoV-2 virus-infected cells was 13.3 nmol·L-1,which might be the main active component of Chansu injection in resisting SARS-CoV-2 virus-infected cells.Chansu injection could significantly inhibit the cell membrane fusion mediated by the S protein of SARS-CoV-2 virus,thereby blocking the virus invasion.CONCLUSION Chansu injection demonstrates in vitro anti-SARS-CoV-2 activity by suppressing NP protein expression,reducing inflammatory cytokine lev-els,and blocking viral entry through membrane fusion inhibition.Bufalin is likely the key active component responsible for its anti-SARS-CoV-2 effects.

OBJECTIVE To investigate the in vitro activity of Chansu injection against SARS-CoV-2 infection and determine whether bufalin is the primary active component responsible for its efficacy.METHODS The half-maximal effective concentration(EC50)of Chansu injection and bufalin against SARS-CoV-2 infection was determined using the CellTiter-Glo cell viability assay.qPCR was performed to assess the effects of Chansu injection and bufalin on mRNA levels of SARS-CoV-2 NP protein,inflammatory factors,and interferons in virus-infected cells.A cell-cell membrane fusion model was established,and the impact of the drugs on membrane fusion between HEK-293T and HEK-293T-ACE2 cells was evaluated using a luciferase reporter gene assay.RESULTS Chansu injection exhibited anti-SARS-CoV-2 virus infection activity,with an EC50 of 85.56 ng·mL-1.Chansu injec-tion significantly reduced the mRNA levels of viral NP protein(P<0.05),IFN-λ2/3(P<0.05),ISG-15 and RIG-I(P<0.000 1)in SARS-CoV-2 infected Calu-3 cells,and the mRNA levels of inflammatory factors IL-6 and TNF-α were significantly reduced(P<0.000 1).The EC50 of bufalin in inhibiting SARS-CoV-2 virus-infected cells was 13.3 nmol·L-1,which might be the main active component of Chansu injection in resisting SARS-CoV-2 virus-infected cells.Chansu injection could significantly inhibit the cell membrane fusion mediated by the S protein of SARS-CoV-2 virus,thereby blocking the virus invasion.CONCLUSION Chansu injection demonstrates in vitro anti-SARS-CoV-2 activity by suppressing NP protein expression,reducing inflammatory cytokine lev-els,and blocking viral entry through membrane fusion inhibition.Bufalin is likely the key active component responsible for its anti-SARS-CoV-2 effects.

Objective:To explore the methods of fertility preservation in prepubertal patients with thalassemia major (TM), and to provide further data support for the fertility preservation in prepubertal patients with hematologic diseases.Methods:A case of a prepubertal patient with TM who required urgent hematopoietic stem cell transplantation (HSCT) receiving ovarian tissue cryopreservation (OTC) and in vitro maturation (IVM) to preserve fertility was reported, and the timing, the indications and strategies of fertility preservation in prepubertal girls with thalassemia were discussed in combination with related literature. Results:After ovarian tissue extraction, a total of 24 cumulus-oocyte complexes (COCs) and 11 ovarian cortex pieces were obtained through puncture and aspiration. After IVM for 48 h, a total of 9 M Ⅱ oocytes were frozen by vitrification. Conclusion:For the prepubertal girls facing HSCT urgently, cryopreserving ovarian tissue in combination with retrieving immature oocytes followed by IVM can preserve the fertility of patients to the greatest extent in a short period of time, as well as improve the therapeutic effect of fertility preservation in patients.

Objective:To explore the methods of fertility preservation in prepubertal patients with thalassemia major (TM), and to provide further data support for the fertility preservation in prepubertal patients with hematologic diseases.Methods:A case of a prepubertal patient with TM who required urgent hematopoietic stem cell transplantation (HSCT) receiving ovarian tissue cryopreservation (OTC) and in vitro maturation (IVM) to preserve fertility was reported, and the timing, the indications and strategies of fertility preservation in prepubertal girls with thalassemia were discussed in combination with related literature. Results:After ovarian tissue extraction, a total of 24 cumulus-oocyte complexes (COCs) and 11 ovarian cortex pieces were obtained through puncture and aspiration. After IVM for 48 h, a total of 9 M Ⅱ oocytes were frozen by vitrification. Conclusion:For the prepubertal girls facing HSCT urgently, cryopreserving ovarian tissue in combination with retrieving immature oocytes followed by IVM can preserve the fertility of patients to the greatest extent in a short period of time, as well as improve the therapeutic effect of fertility preservation in patients.

Objective The scope of this study was aimed to explore the relationship between serum Ghrelin levels and disease severity in non-traumatic ONFH patients. Methods 84 non-traumatic ONFH patients and 84 healthy controls were enrolled for this study in our hospital. The radiographic progression was determined by FICAT grading system. The clinical severity was evaluated by visual analogue scale(VAS),Harris hip scores(HSS). The serum APN concentrations were examined by enzyme-linked immunosorbent assay(ELISA).Serum adiponectin and IL-33 levels were also examined. The relationship between serum Ghrelin levels and radiographic severity,clinical severity and biochemical indices were investigated. Results Serum Ghrelin levels were significantly lower in ONFH patients than in healthy controls. Serum Ghrelin levels were negatively correlated with FICAT grading system. In addition,Serum Ghrelin levels were also negatively related to VAS scores and positively associated with HSS scores ,Last ,Serum Ghrelin levels were positively with adiponectin levels and negatively correlated with IL-33 levels. Conclusion Serum Ghrelin levels in non-traumatic ONFH patients were negatively associated with the disease severity. Decreased serum Ghrelin levels may reflect disease severity of non-traumatic ONFH.

Objective To improve the quality standard of compound Chaihu tea. Methods Pupleuri Radix, Puerariae Lobatae Radix and Forsythiae Fructus were identified by thin layer chromatography (TLC).HPLC method was used to determine the content of puerarin.The determination was performed on a Shimadzu VP-ODS-C18column (250 mm×4.6 mm,5μm) with the mobile phase of methanol-0.2% H3PO4(23:77).The flow rate was 1.0 mL·min-1, the column temperature was set at 30 ℃ and the detection wavelength was 250 nm. Results The TLC spots were clear and well-separated without negative interference.The good linear correlation existed within the range of 1.340 8-56.632 8μg· mL-1 for puerarin (r=0.999 9, n=6) and the average recovery was 102.98% with RSD being 1.48% (n=6). Conclusion The method is simple and accurate.It can be used for quality control of compound Chaihu tea.