Randomized clinical trials are important in both clinical and academic stroke communities with increasing numbers of new design concepts emerging. One of the “less traditional” designs that have gained increasing interest in the last decade is non-inferiority trials. Whilst the concept might appear straightforward, the design and interpretation of non-inferiority trials can be challenging. In this review, we will use exemplars from clinical trials in the stroke field to provide an overview of the advantages and limitations of non-inferiority trials and how they should be interpreted in stroke research.

Randomized clinical trials are important in both clinical and academic stroke communities with increasing numbers of new design concepts emerging. One of the “less traditional” designs that have gained increasing interest in the last decade is non-inferiority trials. Whilst the concept might appear straightforward, the design and interpretation of non-inferiority trials can be challenging. In this review, we will use exemplars from clinical trials in the stroke field to provide an overview of the advantages and limitations of non-inferiority trials and how they should be interpreted in stroke research.

Randomized clinical trials are important in both clinical and academic stroke communities with increasing numbers of new design concepts emerging. One of the “less traditional” designs that have gained increasing interest in the last decade is non-inferiority trials. Whilst the concept might appear straightforward, the design and interpretation of non-inferiority trials can be challenging. In this review, we will use exemplars from clinical trials in the stroke field to provide an overview of the advantages and limitations of non-inferiority trials and how they should be interpreted in stroke research.

Objective To explore the expression of macrophage colony-stimulating factor(MCSF)in oral squamous cell carcinoma(OSCC)and its effects on the proliferation,apoptosis,and migration of OSCC cells.Methods Normal gingival and OSCC tissues were collected,and MCSF protein expression was detected using immunohistochemistry and Western blotting.HSC-4 cells were divided into control(no transfection),shNC(transfection with sequence-free plasmid vector lentivirus),and shMCSF(transfection with silent MCSF plasmid vector lentivirus)groups.The expression of MCSF mRNA and protein in HSC-4 cells was detected using quantitative real-time PCR and Western blotting,respectively.Scratch and Transwell assays were used to detect the migration ability of HSC-4 cells.The TUNEL assay determined the apoptosis ability of HSC-4 cells,while a colony formation assay detected the proliferation ability of HSC-4 cells.Results MCSF was highly expressed in OSCC tissues and HSC-4 cells but weakly expressed in normal gingival tissues and Hacat cells.The migration and proliferation ability of HSC-4 cells in the shMCSF group was lower than that in the shNC group(P＜0.05).The apoptosis ability of HSC-4 cells in the shMCSF group was higher than that in the shNC group(P＜0.05).Conclusion MCSF is upregu-lated in OSCC tissues,promoting cell migration and proliferation,while also reducing the apoptosis of OSCC cells.

Objective:To evaluate the effect of remimazolam tosilate on postoperative delirium (POD) in elderly patients undergoing urological surgery.Methods:In this randomized controlled trial, 220 American Society of Anesthesiologists Physical Status classification Ⅰ-Ⅲ elderly patients of either sex, aged≥65 yr, with body mass index of 18-28 kg/m 2, scheduled for elective urological surgery under general anesthesia, were divided into 2 groups ( n=110 each) using a random number table method: propofol group (group P) and remimazolam tosilate group (group R). Group P received propofol 1.0-1.5 mg/kg for induction and propofol 2-6 mg·kg -1·h -1 for maintenance, while group R received remimazolam tosilate 0.2-0.3 mg/kg for induction and remimazolam tosilate 0.5-1.2 mg·kg -1·h -1 for maintenance. The other drugs for induction and maintenance were the same in the two groups. POD was assessed using the Chinese version of the 3-minute diagnostic interview for Confusion Assessment Method within 3 days after surgery in the two groups. The intraoperative consumption of remifentanil and usage of vasoactive drugs, extubation time, 15-item Quality-of-Recovery scale scores at 24 h after operation, requirement for rescue analgesia within 24 h after operation, and postoperative adverse effects were recorded. Results:There were no significant differences in the incidence of POD within 3 days after operation, 15-item Quality-of-Recovery scale scores at 24 h after operation, or rate of rescue analgesia within 24 h after operation between two groups ( P>0.05). Compared with group P, the requirement for intraoperative vasoactive drugs was significantly reduced, the extubation time was shortened, and the incidence of hypoxemia was decreased within 24 h after operation in group R ( P<0.05 or 0.001). Conclusions:Remimazolam tosilate has no marked effect on the occurrence of POD in elderly patients undergoing urological surgery.

Objective To explore the expression of macrophage colony-stimulating factor(MCSF)in oral squamous cell carcinoma(OSCC)and its effects on the proliferation,apoptosis,and migration of OSCC cells.Methods Normal gingival and OSCC tissues were collected,and MCSF protein expression was detected using immunohistochemistry and Western blotting.HSC-4 cells were divided into control(no transfection),shNC(transfection with sequence-free plasmid vector lentivirus),and shMCSF(transfection with silent MCSF plasmid vector lentivirus)groups.The expression of MCSF mRNA and protein in HSC-4 cells was detected using quantitative real-time PCR and Western blotting,respectively.Scratch and Transwell assays were used to detect the migration ability of HSC-4 cells.The TUNEL assay determined the apoptosis ability of HSC-4 cells,while a colony formation assay detected the proliferation ability of HSC-4 cells.Results MCSF was highly expressed in OSCC tissues and HSC-4 cells but weakly expressed in normal gingival tissues and Hacat cells.The migration and proliferation ability of HSC-4 cells in the shMCSF group was lower than that in the shNC group(P＜0.05).The apoptosis ability of HSC-4 cells in the shMCSF group was higher than that in the shNC group(P＜0.05).Conclusion MCSF is upregu-lated in OSCC tissues,promoting cell migration and proliferation,while also reducing the apoptosis of OSCC cells.

Objective:To evaluate the effect of remimazolam tosilate on postoperative delirium (POD) in elderly patients undergoing urological surgery.Methods:In this randomized controlled trial, 220 American Society of Anesthesiologists Physical Status classification Ⅰ-Ⅲ elderly patients of either sex, aged≥65 yr, with body mass index of 18-28 kg/m 2, scheduled for elective urological surgery under general anesthesia, were divided into 2 groups ( n=110 each) using a random number table method: propofol group (group P) and remimazolam tosilate group (group R). Group P received propofol 1.0-1.5 mg/kg for induction and propofol 2-6 mg·kg -1·h -1 for maintenance, while group R received remimazolam tosilate 0.2-0.3 mg/kg for induction and remimazolam tosilate 0.5-1.2 mg·kg -1·h -1 for maintenance. The other drugs for induction and maintenance were the same in the two groups. POD was assessed using the Chinese version of the 3-minute diagnostic interview for Confusion Assessment Method within 3 days after surgery in the two groups. The intraoperative consumption of remifentanil and usage of vasoactive drugs, extubation time, 15-item Quality-of-Recovery scale scores at 24 h after operation, requirement for rescue analgesia within 24 h after operation, and postoperative adverse effects were recorded. Results:There were no significant differences in the incidence of POD within 3 days after operation, 15-item Quality-of-Recovery scale scores at 24 h after operation, or rate of rescue analgesia within 24 h after operation between two groups ( P>0.05). Compared with group P, the requirement for intraoperative vasoactive drugs was significantly reduced, the extubation time was shortened, and the incidence of hypoxemia was decreased within 24 h after operation in group R ( P<0.05 or 0.001). Conclusions:Remimazolam tosilate has no marked effect on the occurrence of POD in elderly patients undergoing urological surgery.

Objective To study the possibility of salivary microbiota model to diagnose laryngopharyngeal re-flux(LPR).Methods A case-control study was applied to enroll 34 patients as case group who showed significant efficacy after 8 weeks of proton pump inhibitor treatment from February 2022 to November 2022.And 47 healthy volunteers matched by age,gender and body mass index with the case group were enrolled as the control group.Their salivary samples were collected before medication,and the salivary microbiota was detected by 16S rDNA se-quencing.Bioinformatics analysis was conducted on the sequencing results to compare species differences at the ge-nus level.A total of 24 patients and 33 cases in the control group were selected as train set and the rest as test set.Random forest method was used to classify data and ten fold cross validation was applied to select the optimal bacte-rial genus combination to construct a diagnostic model.The probability of disease(POD)index was calculated and receiver operating characteristic curve(ROC)was used to evaluate the diagnostic model in diagnosis of LPR.SPSS 18.0 software was utilized for statistical analysis.Results Compared with the control group,there was a statistical difference in the relative abundance of 22 genera in saliva between the case group and the control group(P＜0.05).A diagnostic model consisting of 6 genera was constructed,namely Lactobacillus,Novosphingobium,Bacillus,Pseudoalteromonas,Ralstonia and Phocaeicola.The area under the ROC curve of the test set was 0.843,the sensi-tivity of the diagnostic model was 60.0％,the specificity was 87.71％,and the Kappa value was 0.470.Conclusion The bacterial combination diagnostic model constructed from saliva microbiota based on microbiome and machine learning can effectively distinguish LPR patients from healthy individuals,which has potential clinical application value.

Objective To investigate the protective effect of 5-hydroxy-6,7-dimethoxyflavone(5-HDF),a compound extracted from Elsholtzia blanda Benth.,against lung injury induced by H1N1 influenza virus and explore its possible mechanism of action.Methods 5-HDF was extracted from Elsholtzia blanda Benth.using ethanol reflux extraction and silica gel chromatography and characterized using NMR and MS analyses.In an A549 cell model of H1N1 influenza virus infection(MOI=0.1),the cytotoxicity of 5-HDF was assessed using MTT assay,and its effect on TRAIL and IL-8 expressions was examined using flow cytometry;Western blotting was used to detect the expression levels of inflammatory,apoptosis,and ferroptosis-related proteins.In a mouse model of H1N1 influenza virus infection established by nasal instillation of 50 μL H1N1 virus at the median lethal dose,the effects of 30 and 60 mg/kg 5-HDF by gavage on body weight,lung index,gross lung anatomy and lung histopathology were observed.Results 5-HDF exhibited no significant cytotoxicity in A549 cells within the concentration range of 0-200 μg/mL.In H1N1-infected A549 cells,treatment with 5-HDF effectively inhibited the activation of phospho-p38 MAPK and phospho-NF-κB p65,lowered the expressions of IL-8,enhanced the expression of anti-ferroptosis proteins(SLC7A11 and GPX4),and inhibited the expressions of apoptosis markers PARP and caspase-3 and the apoptotic factor TRAIL.In H1N1-infected mice,treatment with 5-HDF for 7 days significantly suppressed body weight loss and increment of lung index and obviously alleviated lung tissue pathologies.Conclusion 5-HDF offers protection against H1N1 influenza virus infection in mice possibly by suppressing H1N1-induced ferroptosis,inflammatory responses,and apoptosis via upregulating SLC7A11 and GPX4,inhibiting the activation of phospho-NF-κB p65 and phospho-p38 MAPK,and decreasing the expression of cleaved caspase3 and cleaved PARP.

Objective To investigate the protective effect of 5-hydroxy-6,7-dimethoxyflavone(5-HDF),a compound extracted from Elsholtzia blanda Benth.,against lung injury induced by H1N1 influenza virus and explore its possible mechanism of action.Methods 5-HDF was extracted from Elsholtzia blanda Benth.using ethanol reflux extraction and silica gel chromatography and characterized using NMR and MS analyses.In an A549 cell model of H1N1 influenza virus infection(MOI=0.1),the cytotoxicity of 5-HDF was assessed using MTT assay,and its effect on TRAIL and IL-8 expressions was examined using flow cytometry;Western blotting was used to detect the expression levels of inflammatory,apoptosis,and ferroptosis-related proteins.In a mouse model of H1N1 influenza virus infection established by nasal instillation of 50 μL H1N1 virus at the median lethal dose,the effects of 30 and 60 mg/kg 5-HDF by gavage on body weight,lung index,gross lung anatomy and lung histopathology were observed.Results 5-HDF exhibited no significant cytotoxicity in A549 cells within the concentration range of 0-200 μg/mL.In H1N1-infected A549 cells,treatment with 5-HDF effectively inhibited the activation of phospho-p38 MAPK and phospho-NF-κB p65,lowered the expressions of IL-8,enhanced the expression of anti-ferroptosis proteins(SLC7A11 and GPX4),and inhibited the expressions of apoptosis markers PARP and caspase-3 and the apoptotic factor TRAIL.In H1N1-infected mice,treatment with 5-HDF for 7 days significantly suppressed body weight loss and increment of lung index and obviously alleviated lung tissue pathologies.Conclusion 5-HDF offers protection against H1N1 influenza virus infection in mice possibly by suppressing H1N1-induced ferroptosis,inflammatory responses,and apoptosis via upregulating SLC7A11 and GPX4,inhibiting the activation of phospho-NF-κB p65 and phospho-p38 MAPK,and decreasing the expression of cleaved caspase3 and cleaved PARP.