The influenza A virus (IAV), renowned for its high contagiousness and potential to catalyze global pandemics, poses significant challenges due to the emergence of drug-resistant strains. Given the critical role of RNA polymerase in IAV replication, it stands out as a promising target for anti-IAV therapies. In this study, we identified a novel C-3-substituted oleanolic acid benzyl amide derivative, A5, as a potent inhibitor of the PA_C-PB1_N polymerase subunit interaction, with an IC₅₀ value of 0.96 ± 0.21 μmol/L. A5 specifically targets the highly conserved PA_C domain and demonstrates remarkable efficacy against both laboratory-adapted and clinically isolated IAV strains, including multidrug-resistant strains, with EC₅₀ values ranging from 0.60 to 1.83 μmol/L. Notably, when combined with oseltamivir, A5 exhibits synergistic effects both in vitro and in vivo. In a murine model, dose-dependent administration of A5 leads to a significant reduction in IAV titers, resulting in a high survival rate among treated mice. Additionally, A5 treatment inhibits virus-induced Toll-like receptor 4 activation, attenuates cytokine responses, and protects against IAV-induced inflammatory responses in macrophages. In summary, A5 emerges as a novel inhibitor with high efficiency and broad-spectrum anti-influenza activity.

Objective To study the inhibitory activities of 3-O-β-chacotriosyl glycyrrhetinic acid derivatives against the entry of SARS-CoV-2 into host cells.Methods With pentacyclic triterpene saponin glycyrrhizic acid(a natural SARS-CoV-2 entry inhibitor)as the lead compound,a series of 3-O-β-chacotriosyl glycyrrhetinic acid derivatives were designed and synthesized based on hypridization principle,and their inhibitory activities against virus entry were tested in SARS-CoV-2 pseudovirus-infected cells.The antiviral targets of the lead compound 1b was identified by pseudotyped SARS-CoV-2 infection assay and surface plasmon resonance(SPR)assay,and the S protein-mediated cell-cell fusion assay was used to evaluate the effect of 1b on virus-cell membrane fusion.Molecular docking and single amino acid mutagenesis were carried out to analyze the effect of 1b on binding activitiy of S protein.Results The lead compound 1b showed significant inhibitory effect against Omicron pseudovirus with an EC50 value of 3.28μmol/L(P<0.05),and had broad-spectrum antiviral activity against other SARS-CoV-2 pseudovirus.Spike-dependent cell-cell fusion assay demonstrated an inhibitory effect of 1b against SARS-CoV-2 S protein-mediated cell-cell fusion.Molecular docking analysis predicted that the lead compound 1b could be well fitted into a cavity between the attachment(S1)and fusion(S2)subunits at the 3-fold axis,where it formed multiple hydrophobic interactions with Glu309,Ser305,Arg765 and Lys964 residues with a KD value of-8.6 kcal/mol.The compound 1b at 10,5,2.5 and 1.25 μmol/L showed a significantly reduced inhibitory activity against the pseudovirus with mutated Arg765,Lys964,Glu309 and Leu303(P<0.01).Conclusion 3-O-β-chacotriosyl glycyrrhetinic acid derivatives are capable of stabilizing spike protein in the pre-fusion step to interfere with the fusion of SARS-CoV-2 with host cell membrane,and can thus serve as potential novel small-molecule SARS-CoV-2 fusion inhibitors.

Objective To study the inhibitory activities of 3-O-β-chacotriosyl glycyrrhetinic acid derivatives against the entry of SARS-CoV-2 into host cells.Methods With pentacyclic triterpene saponin glycyrrhizic acid(a natural SARS-CoV-2 entry inhibitor)as the lead compound,a series of 3-O-β-chacotriosyl glycyrrhetinic acid derivatives were designed and synthesized based on hypridization principle,and their inhibitory activities against virus entry were tested in SARS-CoV-2 pseudovirus-infected cells.The antiviral targets of the lead compound 1b was identified by pseudotyped SARS-CoV-2 infection assay and surface plasmon resonance(SPR)assay,and the S protein-mediated cell-cell fusion assay was used to evaluate the effect of 1b on virus-cell membrane fusion.Molecular docking and single amino acid mutagenesis were carried out to analyze the effect of 1b on binding activitiy of S protein.Results The lead compound 1b showed significant inhibitory effect against Omicron pseudovirus with an EC50 value of 3.28μmol/L(P<0.05),and had broad-spectrum antiviral activity against other SARS-CoV-2 pseudovirus.Spike-dependent cell-cell fusion assay demonstrated an inhibitory effect of 1b against SARS-CoV-2 S protein-mediated cell-cell fusion.Molecular docking analysis predicted that the lead compound 1b could be well fitted into a cavity between the attachment(S1)and fusion(S2)subunits at the 3-fold axis,where it formed multiple hydrophobic interactions with Glu309,Ser305,Arg765 and Lys964 residues with a KD value of-8.6 kcal/mol.The compound 1b at 10,5,2.5 and 1.25 μmol/L showed a significantly reduced inhibitory activity against the pseudovirus with mutated Arg765,Lys964,Glu309 and Leu303(P<0.01).Conclusion 3-O-β-chacotriosyl glycyrrhetinic acid derivatives are capable of stabilizing spike protein in the pre-fusion step to interfere with the fusion of SARS-CoV-2 with host cell membrane,and can thus serve as potential novel small-molecule SARS-CoV-2 fusion inhibitors.

Objective To observe the clinical effect of endovascular stent implantation in the treatment of patients with ischemic cerebrovascular disease (ICVD) and carotid artery stenosis. Methods A total of 83 patients with ICVD treatment were selected and divided into observation group with 43 cases and control group with 40 cases, and the hemodynamic indicators and degree of internal carotid artery stenosis, neural markers and vascular endothelial function, degree of neurological deficits, clinical efficacy, and adverse reactions were compared between the two groups. Results Peak systolic velocity (PSV), end-diastolic velocity (EDV), and degree of vascular stenosis in both groups showed a downward trend after treatment, and the declines in PSV, EDV and vascular stenosis in the control group were all significantly smaller than those in the observation group (P < 0.05). After treatment, the positive pentraxin-3 (PTX-3) and neuron-specific enolase (NSE) levels in both groups decreased significantly, while functional movement disorder (FMD) and vascular endothelial growth factor (VEGF) levels increased significantly, and the PTX-3 and NSE levels in the observation group were significantly lower than those in the control group, while the VEGF and FMD levels were significantly higher (P < 0.05). After treatment, scores of the National Institutes of Health Stroke Scale (NIHSS) and the Modified Edinburgh-Scandinavian Stroke Scale (MESSS) in both groups decreased, and the decline in NIHSS score and MESSS in the control group was smaller than that in the observation group. Clinical total effective rate was 80.00% in the control group, which was significantly lower than 97.67% in the observation group (P < 0.05). The incidence of adverse reactions in the observation group was 2.33%, which was significantly lower than 22.50% in the control group (P < 0.05). Conclusion Endovascular stent implantation can effectively improve the clinical efficacy of ICVD patients, relieve the degree of vascular stenosis, improve cognitive function, and has higher safety.

Objective To study the effects and mechanisms of molecular targeted drug combination on multi-driven proliferation hepatocellular carcinoma SK-Hep-1 cells. Methods Four molecular targeted drugs (HG6-64-1, Dasatinib, Crizotinib, and Sunitinib) were used to treat SK-Hep-1 cells, and the monophasic kinetic analysis curve and two-phase analysis curve were drawn. Western blot analysis was used to detect the effects of the above drugs on key signaling pathways in SK-Hep-1 cells. MTT assay was used to detect the effects of the above drugs and their combination on the proliferation of SK-Hep-1 cells. Results Compared with the monophasic kinetic analysis curve, the biphase analysis curve could better fit the effects of molecular targeted drugs on SK-Hep-1 cells, which predicted that the combination of HG6-64-1, Dasatinib, and MK-2206 could effectively inhibit the proliferation of SK-Hep-1 cells. Conclusion Two-phase kinetic analysis can quantitatively describe the response of multi-driven proliferation hepatocellular carcinoma SK-Hep-1 cells to molecular targeted therapy. The combination of HG6-64-1, Dasatinib, and MK-2206 is a potential drug combination for the treatment of hepatocellular carcinoma.

Objective: To investigate the early clinical outcomes of hip anterior short incision and open reduction and internal fixation for the treatment of children with Delbet Ⅱ femoral neck fracture. Methods: A retrospective analysis of 12 children with DelbetⅡ femoral neck fractures was performed with the anterior short incision approach at Department of Pediatric Orthopaedic, Children′s Hospital, Nanjing Medical University from January 2014 to August 2016. There were 7 males and 5 females including 8 cases at left side and 4 cases at right side,aged (11.5±0.8) years (range: 8-14 years).Patients underwent surgery in (12.0±0.9) hours (range: 6-20 hours) after the injury. Displaced fractures were treated by open reduction if closed reduction failed.The surgical incision was performed with the anterior inferior iliac spine centered on, extending along the groin crease, and the length was 3 to 4 cm. The interval between the sartorius and the tensor fascia latae was applied, and the direct and indirect head of rectus femoris were resected to visualize the hip joint capsule. T-shaped incision was made in the capsule. After the reduction was obtained, two or three compression canulated screws were used to fix the fracture site. The healing time, complications and corresponding record outcome by X-ray and clinical examination were recorded. At the latest follow-up,the results were analyzed using the Ratliff criteria and clinical outcomes were assessed by pain, restrictive hip movement, normal activity or the avoidance of games, and the femoral neck in the radiograph. Results: Twelve patients had been followed-up for (30.0±4.1)months(range: 24-36 months). X-ray showed the fractures′ healing time was (8.0±0.4) weeks(range: 7-9 weeks). Ten of the 12 fractures healed after one operation and were associated with no evidence of osteonecrosis of the femoral head. One case occurred premature physeal closure. There were no cases with fracture relocation, coaxvara, delayed union and nonunion or any significant postoperative complications. There was no infection, flexible internal fixation or ruptures. According to Ratliff grade, 9 cases of 12 patients were fine and 3 cases was acceptable. Conclusions Satisfactory short-term effects can be achieved by a mini-incision on the anterior hip for Delbet Ⅱ pediatric femur neck fractures. The anterior incision has the advantage of less trauma, smaller incision scar, better fracture reduction and fixation, and less wound complications.

Objective To investigate the feasibility and clinical effectiveness of fixation with Kirschner wires plus pediatric hip plate (PHP) for femoral neck fracture combined with avulsion fracture of the greater trochanter in children.Methods Between February 2014 and February 2016,6 children were treated by fixation with Kirschner wires plus PHP for femoral neck fracture combined with avulsion fracture of the greater trochanter at Department of Pediatric Orthopaedics,Children's Hospital,Nanjing Medical University.They were 4 boys and 2 girls,aged from 3.5 to 11.1 years (mean,7.2 years).By the Delbet's classification,all the fractures were of type Ⅲ.Clinical efficacy was evaluated according to the Ratliff scoring system at the last follow-ups.Results The intraoperative bleeding ranged from 20 to 50 mL (average,30 mL);the operation time ranged from 45 to 60 min (average,55 min).All the patients were followed up for 12 to 36 months (average,26 months).Bony union was obtained in all cases after an average time of 7 weeks (from 5 to 8 weeks).The Ratliff scores at the last follow-ups showed 5 good and one acceptable cases.No neurovascular lesion or incision infection was observed in the children.Anatomical reduction of the greater trochanter fracture was not achieved in one child but led to no adverse effect.Conclusion Fixation with Kirschner wires plus PHP can successfully treat femoral neck fracture combined with avulsion fracture of the greater trochanter in children.

Objective To investigate the feasibility and clinical effectiveness of modified humeral supracondy-lar step－cut osteotomy(M－HSSCO)for cubitus varus in children.Methods Between June 2016 and June 2017,the study included 25 children (20 boys and 5 girls)presenting cubitus varus at the age of (8.5 ± 0.3)years old (6－11 years old)in the Children's Hospital of Nanjing Medical University.M－HSSCO was used in the osteotomy operation. The initial osteotomy was performed as a right triangle osteotomy.The inferior margin of this right triangle was outlined parallel to the joint line 0.5－1.0 cm above the olecranon fossa.Then,they were incised from the medial end of this parallel line,which made an angle between the first and second lines equal to the desired corrective humerus elbow－wrist angle.Next,from the lateral end of the second line,the third line perpendicular to the first line was cut.Finally, desired right triangle was outlined and removed.The second right triangle osteotomy from proximal line of osteotomy was similar as the above.The medial cortex and periosteum was used as a hinge and rotated to make two osteotomy parts combined steadily.The Bellemore evaluation system was used to evaluate the difference in carrying angle degree be-tween the affected side and normal side after osteotomy,and the different degree of flexion and extension in the elbow joint between preoperation and post－operation.Residual varus deformity and other complications were also evaluated. Results All patients were followed up for 6－18 months (mean 12 months).The preoperative ipsilateral carrying an-gle was－(15°－30°)[－(22.20 ± 1.40)°],the postoperative was 5°－10°[(7. 60 ± 0.47)°]and the final follow－up was 5°－9°[(7.30 ± 0.40)°].There were statistically significant differences in the flexion angles before surgery and after surgery (t＝23.34,P＝0.000).There were statistically significant differences in the flexion angles before surgery and at final follow－up (t＝23.14,P＝0.000).The mean range of joint motion of the elbow was －5°－130° [(132.5 ± 0.5)°]before surgery,0°－125°[(123.6 ± 1.4)°]after surgery and 0°－130°[(126.8 ± 1.3)°]at fi-nal follow－up.The carrying angle reduced did not happen by the final follow－up.There was no statistically significant difference between the flexion angles before surgery and after surgery or by final follow－up(P＞0. 05).Twenty－three (92.0%)patients got an excellent result,2 cases(18.0%)got a good result and no patient (0)got worse according to the criteria described by Bellemore.The first－class incision healing were achieved in all patients.There was no case with postoperative infection,instabilities,vascular and neural injury or any significant postoperative complications. Conclusions M－HSSCO is an effective osteotomy procedure which shows good feasibility and apparent therapeutic effect in treating cubitus varus in children.

Objective To investigate the genotoxicity of aniline and repair dynamics in hepatocytes and lymph-cytes.Methods Aniline was administered intragastrically to SPF Kunming mice ( five mice in each group) in a single dose of 100 mg/kg body weight.The hepatocytes and peripheral blood lymphocytes were obtained at 3, 8, 16, 24, and 32 hours after aniline administration, respectively.The control mice received tap water only.The DNA damages were detected by single cell gel electrophoresis assay ( SCGE) and the time-effect relationship was analyzed.Results The results of SCGE experiment showed that both the tail lenth and tail moment of the hepatocyte DNA were increased gradually from 8 h, and reached the maximum at 16 h ( P<0.01) after aniline administration.As time went on, DNA damage was recovered gradually, and the two DNA damage indexes were completely returned to control levels at 32 h after aniline administration (P>0.05).The two DNA damage indexes of peripheral blood lymphocytes started to increase at 16 h, reached the maxi-mum at 24 h ( P<0.01) , and began to recover at 32 h after aniline administration.Conclusions Our findings suggeste that aniline may be a potential genotoxicant to hepatocytes and peripheral blood lymphocytes.There is a clear time-response relationship in terms of the two DNA damage indexes, indicating that hepatocytes and lymphocytes in mice possess an effi-cient DNA repair mechanism against aniline toxicity.

Aim To study the inhibitory activities of potential new anti-influenza virus agents,3-O-β-chaco-triosyl pentacyclic triterpenoids against the entry of H5N1influenza viruses.Methods Three target com-pounds were designed and synthesized structurally re-lated to the lead compound 3-O-β-chacotriosyl dioscin derivative (1 )with inhibitory activities against H5N1 influenza viruses.The inhibitory activities of these tar-get compounds were tested at a cellular level pseudo vi-rus system targeting H5N1 influenza viruse entry.Re-sults All the compounds 1 a,1 b and 1 c showed po-tent inhibitory activities against the entry of A/Thai-land/Kan353/2004 pseudo virus into the target cells, of which compound 1 b showed the best inhibitory activ-ity with an IC50 value of (1.25 ±0.22)μmol·L-1. Conclusion The SARs analysis of these compounds indicated that replacement of the aglycone moiety of compound 1 with pentacyclic triterpenoids could in-crease antiviral activity.Different types of pentacyclic triterpen as aglycone residue had the significant influ-ence on the inhibitory activity (1 b >1 c >1 a),sug-gesting ursane type of triterpenes was superior to the two other kinds of triterpenes as aglycone residue.