OBJECTIVE To investigate the therapeutic effect of Sanhan Huashi Formula(SHF)on respiratory syncytial virus(RSV)-infected mouse models and explore its potential antiviral and anti-inflammatory mechanisms using lipidomics.METHODS Fifty-four BALB/c mice were randomly divided into six groups(n=9):blank group,model group,Ribavirin group(50 mg·kg-1·d-1),and SHF high(15.46 g·kg-1·d-1),medium(7.73 g·kg-1·d-1),and low-dose(3.87 g·kg-1·d-1)groups.A pneumonia model was established by in-tranasal RSV infection,followed by three consecutive days of oral gavage administration.Lung tissues were collected for histopathologi-cal evaluation using hematoxylin-eosin(HE)staining and inflammation scoring.Real-time quantitative polymerase chain reaction(RT-qPCR)was performed to measure mRNA levels of viral gene fusion protein(F),glycoprotein(G),and inflammatory factors tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)to assess lung viral load and inflammation,while immunofluorescence staining was performed to observe the expression of RSV-F protein in lung tissues.Serum lipidomics analysis was conducted using ultra-performance liquid chromatography-quadrupole-electrostatic field orbitrap high-resolution mass spectrometry(UPLC-Q Exactive Or-bitrap MS)to identify lipid metabolism changes and differential lipids.RESULTS Compared with the blank group,mice in the model group exhibited marked pulmonary inflammatory cell infiltration and tissue injury,with significantly elevated pulmonary histopa-thology scores and lung index.The lung viral load and the mRNA expression levels of the inflammatory factors TNF-α and IL-6 were significantly increased,and immunofluorescence likewise indicated high expression of RSV-F protein in lung tissue.Relative to the model group,treatment with SHF at all tested doses clearly ameliorated lung tissue injury,effectively suppressed viral gene expression and inflammatory cytokine levels,and reduced the fluorescence signal intensity of RSV-F protein in the lungs.Lipidomics analysis re-vealed that compared with the blank group,the model group exhibited marked disturbances in lipid metabolism-characterized by dys-regulation of triacylglycerol(TG),phosphatidylcholine(PC),lysophosphatidylcholine(LPC),sphingomyelin(SM),diacylglycerol(DG),lysophosphatidylethanolamine(LPE),and phosphatidylethanolamine(PE).High-dose SHF treatment reversed these RSV-induced lipid abnormalities.CONCLUSION SHF effectively alleviates RSV-induced pulmonary inflammation and pathological injury,re-duces pulmonary RSV viral load,and may exert these effects by modulating dysregulated lipid metabolism in peripheral blood.

Objective:To investigate the factors affecting the efficacy of 131I treatment for Graves′ disease (GD) and to construct a predictive model for the treatment outcomes of 131I therapy. Methods:Retrospective analysis of the treatment efficacy was performed on 2 190 patients (547 males, 1 643 females, age (42.9±12.4) years) with GD, who received initial 131I treatment in Tianjin Medical University General Hospital between October 2013 and May 2018. Univariate analysis ( χ2 test, et al) and logistic regression were performed to analyze the possible factors affecting the efficacy of 131I treatment. An efficacy prediction model for 131I treatment of GD was constructed, and decision curve analysis (DCA) was used to evaluate the clinical utility of the prediction model. Results:The overall effectiveness rate of 131I treatment for GD patients was 99.95%(2 189/2 190), with a total cure rate of 83.74%(1 834/2 190), among which 94.11%(1 726/1 834) were cured after a single treatment. Pre-treatment thyroid mass was identified as an independent risk factor affecting the efficacy of initial 131I treatment (odds ratio ( OR)=0.983(95% CI: 0.977-0.989), P<0.001). The clinical cure rate was higher in patients who received an adequate dose of 131I compared with that in patients who didn′t receive an adequate dose (79.97%(1 537/1 922) vs 70.52%(189/268); χ2=12.57, P<0.001), but it did not increase the incidence of hypothyroidism within one year. A predictive model was constructed, and it was found that thyroid mass and disease duration had a relatively high impact on the clinical cure rate. The concordance index (C-index) of the predictive model was 0.623(95% CI: 0.593-0.654). DCA indicated that the predictive model offered substantial net benefits across a wide range of probability thresholds. Conclusions:131I treatment is effective in most patients with GD. The predictive model for efficacy of initial 131I treatment developed in this study can assist in evaluating treatment outcomes and help clinicians select the most suitable 131I treatment dose, enhancing clinical decision-making.

Objective·To explore the efficacy of the XP-Endo Shaper(XPS)and Reciproc(RC)engine-driven nickel-titanium file systems in removing root canal fillings,and to provide evidence for clinical application.Methods·The maxillary first permanent molars were obtained from patients who visited the Department of General Dentistry,Shanghai Ninth People's Hospital,Shanghai Jiao Tong University School of Medicine,between March and September 2022,which required tooth extraction due to periodontal disease.A total of 30 mesiobuccal or distobuccal root canals with moderate curvature were selected and randomly divided into the RC file group(control group,n=15)and the XPS file group(experimental group,n=15).All root canals were first negotiated and shaped using M3-PRO nickel-titanium files,followed by root canal obturation with bioceramic paste using the single-cone technique.Then,RC files(R25)were used in the control group,while XPS files were used in the experimental group to remove the root canal obturation.Cone Beam Computed Tomography(CBCT)scans were performed before and after the removal procedures.The time required for obturation removal,the percentage of residual obturation volume,the amount of apical debris extrusion,and the degree of instrument deformation were recorded.Independent-sample t-tests and chi-square test were used to compare the differences between the two groups.Results·The XPS group required slightly more time(118.87±18.58)s than the RC group(107.93±14.79)s,but the difference between two groups was not statistically significant.The XPS group had a lower percentage of residual obturation volume(7.51±8.06)%compared to the RC group(15.02±14.63)%,but the difference was not statistically significant.The XPS group had significantly less apical debris extrusion(6.15±1.42)mg than the RC group(8.29±2.01)mg,with a statistically significant difference(P=0.002).Both groups exhibited varying degrees of instrument deformation,but yet no instrument separation occurred in either group.Conclusion·The RC engine-driven nickel-titanium files were slightly more time-efficient in removing root canal fillings,whereas the XPS files achieved a higher canal cleanliness and produced less apical debris extrusion.As there was no significant difference in removal time,and considering the superior cleaning performance and reduced risk of postoperative complications,the XPS file system appears more suitable for removing root canal fillings in moderately curved root canals.

Objective To investigate the mechanism of benzyl isothiocyanate(BITC)combined with sorafenib(Sor)in the treatment of anaplastic thyroid cancer(ATC).Methods Two ATC cell lines,8505C and CAL-62,were treated with Sor at concentrations of 0,20,30,40,and 50 μmol/L.The cell survival rate was assessed using CCK-8 assay.The combined dose of BITC and Sor was determined by calculating combination index(CI).CAL-62 and 8505C cells were exposed to 10 μmol/L BITC(BITC group),10 μmol/L Sor(Sor group),or a combination of 10 μmol/L BITC and 10 μmol/L Sor(BITC+Sor group)for 24 hours.The control group was not treated.The effects of Sor and BITC on ATC cell viability were evaluated using the CCK-8 method.Apoptosis was analyzed via flow cytometry.Western blot assay was employed to detect the protein expression levels of LC3B Ⅱ,Beclin-1 and nuclear factor(NF)-κB.Real-time fluorescence quantitative PCR was used to quantify the mRNA levels of LC3B.Additionally,CAL-62 cells were subcutaneously injected into mice to establish tumor xenograft model.Mice were treated with BITC(100 mg/kg,intraperitoneal injection),Sor(30 mg/kg,intragastric administration)or a combination of BITC and Sor every other day for 21 days.Finally,the expression levels of LC3B Ⅱ,Beclin-1 and NF-κB in tumor tissue were analyzed by Western blot assay.Results Sor significantly inhibited the viability of CAL-62 and 8505C cells in a concentration-dependent manner.The combination index(CI)was 0.710 at BITC 10 μmol/L and Sor 10 μmol/L,indicating a moderate synergistic effect between the two drugs.In both 8505C and CAL-62 cells,compared with the control group,treatment with BITC or Sor resulted in the decreased cell viability,as well as reduced expression levels of Beclin-1 and NF-κB proteins(P＜0.05),and the apoptosis rate,LC3B mRNA and LC3B Ⅱ protein expression levels were significantly increased(P＜0.05).When BITC and Sor were combined,the cell viability,Beclin-1 and NF-κB protein expressions were further reduced compared to either drug alone,while the apoptosis rate,LC3B mRNA and LC3B Ⅱ protein expression levels were significantly elevated(P＜0.05).In the mouse xenograft tumor model,the BITC+Sor group exhibited increased LC3B Ⅱ expression,along with decreased Beclin-1 and NF-κB expression levels,tumor volume and tumor mass compared to the BITC or Sor groups(P＜0.05).Conclusion The combination of BITC and Sor can inhibit ATC cells through NF-κB pathway,induce autophagy and promote apoptosis in vitro and in vivo.

Objective·To explore the efficacy of the XP-Endo Shaper(XPS)and Reciproc(RC)engine-driven nickel-titanium file systems in removing root canal fillings,and to provide evidence for clinical application.Methods·The maxillary first permanent molars were obtained from patients who visited the Department of General Dentistry,Shanghai Ninth People's Hospital,Shanghai Jiao Tong University School of Medicine,between March and September 2022,which required tooth extraction due to periodontal disease.A total of 30 mesiobuccal or distobuccal root canals with moderate curvature were selected and randomly divided into the RC file group(control group,n=15)and the XPS file group(experimental group,n=15).All root canals were first negotiated and shaped using M3-PRO nickel-titanium files,followed by root canal obturation with bioceramic paste using the single-cone technique.Then,RC files(R25)were used in the control group,while XPS files were used in the experimental group to remove the root canal obturation.Cone Beam Computed Tomography(CBCT)scans were performed before and after the removal procedures.The time required for obturation removal,the percentage of residual obturation volume,the amount of apical debris extrusion,and the degree of instrument deformation were recorded.Independent-sample t-tests and chi-square test were used to compare the differences between the two groups.Results·The XPS group required slightly more time(118.87±18.58)s than the RC group(107.93±14.79)s,but the difference between two groups was not statistically significant.The XPS group had a lower percentage of residual obturation volume(7.51±8.06)%compared to the RC group(15.02±14.63)%,but the difference was not statistically significant.The XPS group had significantly less apical debris extrusion(6.15±1.42)mg than the RC group(8.29±2.01)mg,with a statistically significant difference(P=0.002).Both groups exhibited varying degrees of instrument deformation,but yet no instrument separation occurred in either group.Conclusion·The RC engine-driven nickel-titanium files were slightly more time-efficient in removing root canal fillings,whereas the XPS files achieved a higher canal cleanliness and produced less apical debris extrusion.As there was no significant difference in removal time,and considering the superior cleaning performance and reduced risk of postoperative complications,the XPS file system appears more suitable for removing root canal fillings in moderately curved root canals.

OBJECTIVE To investigate the therapeutic effect of Sanhan Huashi Formula(SHF)on respiratory syncytial virus(RSV)-infected mouse models and explore its potential antiviral and anti-inflammatory mechanisms using lipidomics.METHODS Fifty-four BALB/c mice were randomly divided into six groups(n=9):blank group,model group,Ribavirin group(50 mg·kg-1·d-1),and SHF high(15.46 g·kg-1·d-1),medium(7.73 g·kg-1·d-1),and low-dose(3.87 g·kg-1·d-1)groups.A pneumonia model was established by in-tranasal RSV infection,followed by three consecutive days of oral gavage administration.Lung tissues were collected for histopathologi-cal evaluation using hematoxylin-eosin(HE)staining and inflammation scoring.Real-time quantitative polymerase chain reaction(RT-qPCR)was performed to measure mRNA levels of viral gene fusion protein(F),glycoprotein(G),and inflammatory factors tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)to assess lung viral load and inflammation,while immunofluorescence staining was performed to observe the expression of RSV-F protein in lung tissues.Serum lipidomics analysis was conducted using ultra-performance liquid chromatography-quadrupole-electrostatic field orbitrap high-resolution mass spectrometry(UPLC-Q Exactive Or-bitrap MS)to identify lipid metabolism changes and differential lipids.RESULTS Compared with the blank group,mice in the model group exhibited marked pulmonary inflammatory cell infiltration and tissue injury,with significantly elevated pulmonary histopa-thology scores and lung index.The lung viral load and the mRNA expression levels of the inflammatory factors TNF-α and IL-6 were significantly increased,and immunofluorescence likewise indicated high expression of RSV-F protein in lung tissue.Relative to the model group,treatment with SHF at all tested doses clearly ameliorated lung tissue injury,effectively suppressed viral gene expression and inflammatory cytokine levels,and reduced the fluorescence signal intensity of RSV-F protein in the lungs.Lipidomics analysis re-vealed that compared with the blank group,the model group exhibited marked disturbances in lipid metabolism-characterized by dys-regulation of triacylglycerol(TG),phosphatidylcholine(PC),lysophosphatidylcholine(LPC),sphingomyelin(SM),diacylglycerol(DG),lysophosphatidylethanolamine(LPE),and phosphatidylethanolamine(PE).High-dose SHF treatment reversed these RSV-induced lipid abnormalities.CONCLUSION SHF effectively alleviates RSV-induced pulmonary inflammation and pathological injury,re-duces pulmonary RSV viral load,and may exert these effects by modulating dysregulated lipid metabolism in peripheral blood.

Objective:To investigate the factors affecting the efficacy of 131I treatment for Graves′ disease (GD) and to construct a predictive model for the treatment outcomes of 131I therapy. Methods:Retrospective analysis of the treatment efficacy was performed on 2 190 patients (547 males, 1 643 females, age (42.9±12.4) years) with GD, who received initial 131I treatment in Tianjin Medical University General Hospital between October 2013 and May 2018. Univariate analysis ( χ2 test, et al) and logistic regression were performed to analyze the possible factors affecting the efficacy of 131I treatment. An efficacy prediction model for 131I treatment of GD was constructed, and decision curve analysis (DCA) was used to evaluate the clinical utility of the prediction model. Results:The overall effectiveness rate of 131I treatment for GD patients was 99.95%(2 189/2 190), with a total cure rate of 83.74%(1 834/2 190), among which 94.11%(1 726/1 834) were cured after a single treatment. Pre-treatment thyroid mass was identified as an independent risk factor affecting the efficacy of initial 131I treatment (odds ratio ( OR)=0.983(95% CI: 0.977-0.989), P<0.001). The clinical cure rate was higher in patients who received an adequate dose of 131I compared with that in patients who didn′t receive an adequate dose (79.97%(1 537/1 922) vs 70.52%(189/268); χ2=12.57, P<0.001), but it did not increase the incidence of hypothyroidism within one year. A predictive model was constructed, and it was found that thyroid mass and disease duration had a relatively high impact on the clinical cure rate. The concordance index (C-index) of the predictive model was 0.623(95% CI: 0.593-0.654). DCA indicated that the predictive model offered substantial net benefits across a wide range of probability thresholds. Conclusions:131I treatment is effective in most patients with GD. The predictive model for efficacy of initial 131I treatment developed in this study can assist in evaluating treatment outcomes and help clinicians select the most suitable 131I treatment dose, enhancing clinical decision-making.

Objective:To investigate the inpact of thyroid cancer-derived cancer-associated fibroblasts(CAF) autophagy on papillary thyroid cancer(PTC).Methods:CAF and normal fibroblasts were isolated from cancerous and adjacent normal thyroid tissues from four PTC patients. Expressions of fibroblast activation protein(FAP) and α-smooth muscle actin in cells were assessed. Conditioned medium of CAF and normal fibroblasts were prepared and used to culture PTC cells. The effects of CAF and normal fibroblasts on survival, proliferation, migration, invasion and iodine uptake of PTC cells were evaluated through cell proliferation assay, cell scratch assay, cell invasion assay, and cell iodine uptake assay. The autophagy level of CAF was also evaluated. Autophagy inhibition and activation were used to regulate the autophagy of CAF, and then their effects on PTC cell proliferation, migration and invasion were further evaluated. The in vivo effect of CAF autophagy on PTC xenograft tumor growth was evaluated.Results:CAF exhibited higher FAP expression and basal autophagy levels. PTC cells co-cultured with CAF-conditioned media showed enhanced proliferation, migration, invasion, and reduced iodine uptake. Autophagy inhibition reduced these effects, while autophagy activation further promoted them. In vivo, inhibiting CAF autophagy suppressed tumor growth.Conclusions:CAF promotes PTC cell malignancy through autophagy activation, enhancing proliferation, migration, and invasion while reducing iodine uptake.

Objective:To investigate the inpact of thyroid cancer-derived cancer-associated fibroblasts(CAF) autophagy on papillary thyroid cancer(PTC).Methods:CAF and normal fibroblasts were isolated from cancerous and adjacent normal thyroid tissues from four PTC patients. Expressions of fibroblast activation protein(FAP) and α-smooth muscle actin in cells were assessed. Conditioned medium of CAF and normal fibroblasts were prepared and used to culture PTC cells. The effects of CAF and normal fibroblasts on survival, proliferation, migration, invasion and iodine uptake of PTC cells were evaluated through cell proliferation assay, cell scratch assay, cell invasion assay, and cell iodine uptake assay. The autophagy level of CAF was also evaluated. Autophagy inhibition and activation were used to regulate the autophagy of CAF, and then their effects on PTC cell proliferation, migration and invasion were further evaluated. The in vivo effect of CAF autophagy on PTC xenograft tumor growth was evaluated.Results:CAF exhibited higher FAP expression and basal autophagy levels. PTC cells co-cultured with CAF-conditioned media showed enhanced proliferation, migration, invasion, and reduced iodine uptake. Autophagy inhibition reduced these effects, while autophagy activation further promoted them. In vivo, inhibiting CAF autophagy suppressed tumor growth.Conclusions:CAF promotes PTC cell malignancy through autophagy activation, enhancing proliferation, migration, and invasion while reducing iodine uptake.

Objective To investigate the mechanism of benzyl isothiocyanate(BITC)combined with sorafenib(Sor)in the treatment of anaplastic thyroid cancer(ATC).Methods Two ATC cell lines,8505C and CAL-62,were treated with Sor at concentrations of 0,20,30,40,and 50 μmol/L.The cell survival rate was assessed using CCK-8 assay.The combined dose of BITC and Sor was determined by calculating combination index(CI).CAL-62 and 8505C cells were exposed to 10 μmol/L BITC(BITC group),10 μmol/L Sor(Sor group),or a combination of 10 μmol/L BITC and 10 μmol/L Sor(BITC+Sor group)for 24 hours.The control group was not treated.The effects of Sor and BITC on ATC cell viability were evaluated using the CCK-8 method.Apoptosis was analyzed via flow cytometry.Western blot assay was employed to detect the protein expression levels of LC3B Ⅱ,Beclin-1 and nuclear factor(NF)-κB.Real-time fluorescence quantitative PCR was used to quantify the mRNA levels of LC3B.Additionally,CAL-62 cells were subcutaneously injected into mice to establish tumor xenograft model.Mice were treated with BITC(100 mg/kg,intraperitoneal injection),Sor(30 mg/kg,intragastric administration)or a combination of BITC and Sor every other day for 21 days.Finally,the expression levels of LC3B Ⅱ,Beclin-1 and NF-κB in tumor tissue were analyzed by Western blot assay.Results Sor significantly inhibited the viability of CAL-62 and 8505C cells in a concentration-dependent manner.The combination index(CI)was 0.710 at BITC 10 μmol/L and Sor 10 μmol/L,indicating a moderate synergistic effect between the two drugs.In both 8505C and CAL-62 cells,compared with the control group,treatment with BITC or Sor resulted in the decreased cell viability,as well as reduced expression levels of Beclin-1 and NF-κB proteins(P＜0.05),and the apoptosis rate,LC3B mRNA and LC3B Ⅱ protein expression levels were significantly increased(P＜0.05).When BITC and Sor were combined,the cell viability,Beclin-1 and NF-κB protein expressions were further reduced compared to either drug alone,while the apoptosis rate,LC3B mRNA and LC3B Ⅱ protein expression levels were significantly elevated(P＜0.05).In the mouse xenograft tumor model,the BITC+Sor group exhibited increased LC3B Ⅱ expression,along with decreased Beclin-1 and NF-κB expression levels,tumor volume and tumor mass compared to the BITC or Sor groups(P＜0.05).Conclusion The combination of BITC and Sor can inhibit ATC cells through NF-κB pathway,induce autophagy and promote apoptosis in vitro and in vivo.