Objective To develop an ICU patient care difficulty index based on psychosocial factors and to evaluate its reliability and validity,thereby providing a comprehensive tool for assessing the difficulties in ICU patient care.Methods Guided by Guarinoni theory,an index system was developed through systematic literature reviews,semi-structured interviews,two rounds of Delphi expert consultation and the analytic hierarchy process.Based on the drafted system,a questionnaire was formulated.The tests for validity and reliability were conducted on 290 patients selected by convenience sampling.Results The finalised ICU patient care difficulty index system was composed of 5 primary indices(the general condition of the patient,disease condition,nursing condition,social support,and organizational characteristics),13 secondary indices and 27 tertiary indices.The Kendall's coefficient of coordination W values were 0.380 and 0.498,respectively,indicating a statistically significant agreement(both P<0.001).The system demonstrated a robust reliability(Cronbach's α=0.896)and a good internal consistency.Validity was confirmed through a scale level-content validity index(S-CVI)of 0.94 and the item level-content validity indices(I-CVI)ranged from 0.87 to 1.00.Structural validity analysis,based on 8 extracted public factors,showed a cumulative variance contribution rate of 66.34%.Conclusion The ICU patient care difficulty index system shows a high reliability and validity,making it a valuable tool for accurately quantification of the difficulty in ICU patient care.This system provides a scientific basis for allocation of ICU nursing resource and performance distribution.

Objective:To explore the safety and efficacy of intraosseous regional administration (IORA) of vancomycin for preventing infection in primary total knee arthroplasty (TKA).Methods:A total of 124 patients with knee osteoarthritis undergoing TKA between February 2024 and May 2024 at nine hospitals were enrolled. Preoperative infection prophylaxis involved either IORA (0.5 g vancomycin administered via intraosseous regional infusion before incision) or intravenous infusion (1 g vancomycin via peripheral vein). The IORA group included 15 males and 47 females with a median age of 66.5 years (range, 60.0-70.0 years), while the intravenous group included 14 males and 48 females with a median age of 66.0 years (range, 61.8-70.3 years) years. Intraoperative samples were collected including fat and synovium tissues after incision, before prosthesis placement, and after tourniquet release; distal femoral cancellous bone during femoral osteotomy; proximal tibial cancellous bone during tibial osteotomy; proximal intercondylar cancellous bone before prosthesis placement; and peripheral blood from non-infused arms at surgery initiation and after tourniquet release. Vancomycin concentrations were measured using liquid chromatography-tandem mass spectrometry. Vital sign changes were recorded from admission to 5~10 minutes post-IORA (IORA group) or post-incision (intravenous group). Follow-ups were conducted on postoperative day 1 and 3, and at 1 and 3 months, to document complications including IORA-related adverse events, periprosthetic joint infections, surgical site infections, red man syndrome, acute kidney injury, deep vein thrombosis and so on.Results:Vancomycin concentrations in bone, fat, and synovial tissue samples were significantly higher in the IORA group than in the intravenous group ( P<0.05), while vancomycin concentrations in blood samples were significantly lower in the IORA group than in the intravenous group ( P<0.05). Only 7.3%(41/558) of tissue samples in the IORA group had vancomycin concentrations below 2.0 μg/g (the minimum inhibitory concentration of vancomycin against coagulase-negative staphylococcus), compared to 59.3%(331/558) in the intravenous group (χ 2=11.285, P<0.001). In the intravenous group, 16.9%(21/124) of blood samples had vancomycin concentrations exceeding 15.0 mg/L (the threshold associated with a significantly increased risk of nephrotoxicity), while all concentrations in the IORA group were below this threshold, the difference was statistically significant (χ 2=22.943, P<0.001). There were no statistically significant difference ( P>0.05) in vital signs changes before and after vancomycin administration between the two groups. Two patients in the intravenous group experienced incision exudate, while no other related complications occurred in either group. Conclusions:Compared to the traditional intravenous infusion of 1 g vancomycin, intraosseous injection of a low dose (0.5 g) of vancomycin achieves higher local tissue concentrations in the knee joint with a lower incidence of adverse reactions and is safe for infection prophylaxis. Despite guidelines not recommending the routine use of vancomycin for preventing infection after primary TKA, intraosseous injection of 0.5 g vancomycin may be considered intraoperatively for primary TKA in the following scenarios: patients in medical institutions with a high prevalence of methicillin-resistant staphylococcus aureus (MRSA) infections, patients with potential preoperative MRSA colonization, or patients with cephalosporin allergy.

The objective of this study was to develop a rapid and precise detection technique for por-cine Senecavirus A(SVA)employing reverse transcription recombinase polymerase amplification(RT-RAA)in conjunction with CRISPR Cas13a technology.Additionally,the study aimed to opti-mize the assay's reaction conditions to enhance amplification efficiency.Eight RT-RAA primer sets were designed based on the conserved gene sequence of porcine SVA,and a series of reaction condi-tions were evaluated to refine the RT-RAA reaction system.Subsequently,CRISPR-derived RNA(crRNA)sequences were developed and selected to construct the RT-RAA-CRISPR reaction sys-tem.The method's specificity was determined by examining six prevalent porcine pathogenic nucleic acids,while its sensitivity was assessed using SVA cRNA standards quantified by digital PCR.The method's stability and the consistency of clinical sample analysis were also evaluated.The findings revealed that the optimized RT-RAA and CRISPR reaction systems exhibited the highest amplifi-cation efficiency at a reaction temperature of 37 ℃.Among the eight crRNAs,five were identified as exhibiting the strongest detection signals.The formulated RT-RAA-CRISPR Cas13a method demonstrated exceptional specificity,showing no cross-reactivity with other common porcine disea-ses,including ASFV,PRRSV,PEDV,PCV2,CSFV,and PRV.The method achieved high sensitivi-ty,detecting as low as 0.86 copies/μL of SVA,and exhibited stable fluorescence output,robust re-producibility,and the ability to complete clinical sample analysis within 50 minutes.Consistency e-valuation with six positive and 58 negative samples indicated 100％agreement in outcomes.These results substantiate that the study successfully established a rapid and specific RT-RAA-CRISPR Cas13a detection method for the on-site identification of porcine Senecavirus A,demonstrating high specificity and sensitivity,and holds promise for application in SVA monitoring and control initia-tives.

Objective:To explore the safety and efficacy of intraosseous regional administration (IORA) of vancomycin for preventing infection in primary total knee arthroplasty (TKA).Methods:A total of 124 patients with knee osteoarthritis undergoing TKA between February 2024 and May 2024 at nine hospitals were enrolled. Preoperative infection prophylaxis involved either IORA (0.5 g vancomycin administered via intraosseous regional infusion before incision) or intravenous infusion (1 g vancomycin via peripheral vein). The IORA group included 15 males and 47 females with a median age of 66.5 years (range, 60.0-70.0 years), while the intravenous group included 14 males and 48 females with a median age of 66.0 years (range, 61.8-70.3 years) years. Intraoperative samples were collected including fat and synovium tissues after incision, before prosthesis placement, and after tourniquet release; distal femoral cancellous bone during femoral osteotomy; proximal tibial cancellous bone during tibial osteotomy; proximal intercondylar cancellous bone before prosthesis placement; and peripheral blood from non-infused arms at surgery initiation and after tourniquet release. Vancomycin concentrations were measured using liquid chromatography-tandem mass spectrometry. Vital sign changes were recorded from admission to 5~10 minutes post-IORA (IORA group) or post-incision (intravenous group). Follow-ups were conducted on postoperative day 1 and 3, and at 1 and 3 months, to document complications including IORA-related adverse events, periprosthetic joint infections, surgical site infections, red man syndrome, acute kidney injury, deep vein thrombosis and so on.Results:Vancomycin concentrations in bone, fat, and synovial tissue samples were significantly higher in the IORA group than in the intravenous group ( P<0.05), while vancomycin concentrations in blood samples were significantly lower in the IORA group than in the intravenous group ( P<0.05). Only 7.3%(41/558) of tissue samples in the IORA group had vancomycin concentrations below 2.0 μg/g (the minimum inhibitory concentration of vancomycin against coagulase-negative staphylococcus), compared to 59.3%(331/558) in the intravenous group (χ 2=11.285, P<0.001). In the intravenous group, 16.9%(21/124) of blood samples had vancomycin concentrations exceeding 15.0 mg/L (the threshold associated with a significantly increased risk of nephrotoxicity), while all concentrations in the IORA group were below this threshold, the difference was statistically significant (χ 2=22.943, P<0.001). There were no statistically significant difference ( P>0.05) in vital signs changes before and after vancomycin administration between the two groups. Two patients in the intravenous group experienced incision exudate, while no other related complications occurred in either group. Conclusions:Compared to the traditional intravenous infusion of 1 g vancomycin, intraosseous injection of a low dose (0.5 g) of vancomycin achieves higher local tissue concentrations in the knee joint with a lower incidence of adverse reactions and is safe for infection prophylaxis. Despite guidelines not recommending the routine use of vancomycin for preventing infection after primary TKA, intraosseous injection of 0.5 g vancomycin may be considered intraoperatively for primary TKA in the following scenarios: patients in medical institutions with a high prevalence of methicillin-resistant staphylococcus aureus (MRSA) infections, patients with potential preoperative MRSA colonization, or patients with cephalosporin allergy.

Objective To explore the value of machine learning(ML)models based on ultrasonic texture features(TF)of coronary artery for predicting incomplete Kawasaki disease(IKD)in children.Methods Forty-eight children with IKD and 48 children without KD(non-KD)were enrolled with propensity score matching and divided into training set(n=67,34 cases of IKD and 33 cases of non-KD)and test set(n=29,14 of IKD and 15 of non-KD)at the ratio of 7∶3.Based on clinic-laboratory indicators(C-L)in training set and TF obtained with texture analysis of coronary artery ultrasound images,the optimal C-L-related features and TF were selected.Based on the optimal C-L correlated features,TF and their combinations,6 ML models,including random forest(RF),support vector machine(SVM),logistic regression(LR),gradient boosting decision tree(GBDT),decision tree(DT)and eXtreme gradient boosting(XGBoost)were respectively constructed for predicting IKD in children.The models were then trained in training set and validated in test set,and the best C-L ML,TF ML and C-L-TF ML models were selected.The area under the curve(AUC)of the best ML models were compared,and the clinical value of the best TF ML model was observed with decision curve analysis(DCA).Results Totally 3 optimal C-L related features and 8 optimal TF were selected.Among the constructed C-L ML,TF ML and C-L-TF ML models,C-L-LR model,TF-LR model and C-L-TF-SVM model were the optimal ones,with AUC in training set of 0.891,0.985 and 0.965,while in test set of 0.676,0.971 and 0.948,respectively.No significant difference of AUC was found between TF-LR model and C-L-TF-SVM model in both training set and test set(both P＞0.05),which were both greater than those of C-L-LR model(all P＜0.05).TF-LR model achieved higher clinical benefits in both training set and test set.Conclusion Ultrasound TF-LR model of coronary artery could be used to effectively predict IKD in children.

Objective:To investigate the effect of exosomes released from triple-negative breast cancer cells (TNBC) on immune cells after radiotherapy.Methods:When TNBC (3 types: MDA-MB-231, MDA-MB-453, MDA-MB-468) reached 70% confluence, cells were irradiated with a dose of 8 Gy in one group and no intervention was given in the control group. The supernatants were collected at 48 h after irradiation. Subsequently, these supernatants were co-cultured with lymphocytes in peripheral blood mononuclear cells, and the uptake of exosomes by T cells was confirmed by fluorescence microscopy. Meanwhile, the expression of regulatory T cells (Treg) in the cells was detected using flow cytometry. Differences in Treg cell differentiation between two groups were compared by t-test (expressed as Treg cell positivity rate). Results:Transmission electron microscopy scanning and nanoparticle analysis showed that the extracellular vesicles extracted in the experiment were exosomes. Lymphocytes phagocytosed the exosomes and the exosomes were mainly concentrated in the cytoplasm after phagocytosis. Following the co-culture of 3 kinds of lymphocytes with exosomes from TNBC, there was an increase in Treg cell differentiation compared to control group (1.07%, 0.60%, 0.63% vs. 0.30%, P<0.01). In addition, exosomes released from TNBC further increased the differentiation of Treg cells after radiotherapy (MDA-MB-231 cells: 1.07% vs. 1.81%, P<0.01; MDA-MB-453 cells: 0.60% vs. 0.93%, P<0.05). Conclusions:In summary, exosomes released from TNBC can promote the differentiation of Treg cells. In addition, radiotherapy-induced exosomes released by TNBC further exacerbate the differentiation of Treg cell.

Objective To develop an ICU patient care difficulty index based on psychosocial factors and to evaluate its reliability and validity,thereby providing a comprehensive tool for assessing the difficulties in ICU patient care.Methods Guided by Guarinoni theory,an index system was developed through systematic literature reviews,semi-structured interviews,two rounds of Delphi expert consultation and the analytic hierarchy process.Based on the drafted system,a questionnaire was formulated.The tests for validity and reliability were conducted on 290 patients selected by convenience sampling.Results The finalised ICU patient care difficulty index system was composed of 5 primary indices(the general condition of the patient,disease condition,nursing condition,social support,and organizational characteristics),13 secondary indices and 27 tertiary indices.The Kendall's coefficient of coordination W values were 0.380 and 0.498,respectively,indicating a statistically significant agreement(both P<0.001).The system demonstrated a robust reliability(Cronbach's α=0.896)and a good internal consistency.Validity was confirmed through a scale level-content validity index(S-CVI)of 0.94 and the item level-content validity indices(I-CVI)ranged from 0.87 to 1.00.Structural validity analysis,based on 8 extracted public factors,showed a cumulative variance contribution rate of 66.34%.Conclusion The ICU patient care difficulty index system shows a high reliability and validity,making it a valuable tool for accurately quantification of the difficulty in ICU patient care.This system provides a scientific basis for allocation of ICU nursing resource and performance distribution.

The objective of this study was to develop a rapid and precise detection technique for por-cine Senecavirus A(SVA)employing reverse transcription recombinase polymerase amplification(RT-RAA)in conjunction with CRISPR Cas13a technology.Additionally,the study aimed to opti-mize the assay's reaction conditions to enhance amplification efficiency.Eight RT-RAA primer sets were designed based on the conserved gene sequence of porcine SVA,and a series of reaction condi-tions were evaluated to refine the RT-RAA reaction system.Subsequently,CRISPR-derived RNA(crRNA)sequences were developed and selected to construct the RT-RAA-CRISPR reaction sys-tem.The method's specificity was determined by examining six prevalent porcine pathogenic nucleic acids,while its sensitivity was assessed using SVA cRNA standards quantified by digital PCR.The method's stability and the consistency of clinical sample analysis were also evaluated.The findings revealed that the optimized RT-RAA and CRISPR reaction systems exhibited the highest amplifi-cation efficiency at a reaction temperature of 37 ℃.Among the eight crRNAs,five were identified as exhibiting the strongest detection signals.The formulated RT-RAA-CRISPR Cas13a method demonstrated exceptional specificity,showing no cross-reactivity with other common porcine disea-ses,including ASFV,PRRSV,PEDV,PCV2,CSFV,and PRV.The method achieved high sensitivi-ty,detecting as low as 0.86 copies/μL of SVA,and exhibited stable fluorescence output,robust re-producibility,and the ability to complete clinical sample analysis within 50 minutes.Consistency e-valuation with six positive and 58 negative samples indicated 100％agreement in outcomes.These results substantiate that the study successfully established a rapid and specific RT-RAA-CRISPR Cas13a detection method for the on-site identification of porcine Senecavirus A,demonstrating high specificity and sensitivity,and holds promise for application in SVA monitoring and control initia-tives.

Objective To explore the value of machine learning(ML)models based on ultrasonic texture features(TF)of coronary artery for predicting incomplete Kawasaki disease(IKD)in children.Methods Forty-eight children with IKD and 48 children without KD(non-KD)were enrolled with propensity score matching and divided into training set(n=67,34 cases of IKD and 33 cases of non-KD)and test set(n=29,14 of IKD and 15 of non-KD)at the ratio of 7∶3.Based on clinic-laboratory indicators(C-L)in training set and TF obtained with texture analysis of coronary artery ultrasound images,the optimal C-L-related features and TF were selected.Based on the optimal C-L correlated features,TF and their combinations,6 ML models,including random forest(RF),support vector machine(SVM),logistic regression(LR),gradient boosting decision tree(GBDT),decision tree(DT)and eXtreme gradient boosting(XGBoost)were respectively constructed for predicting IKD in children.The models were then trained in training set and validated in test set,and the best C-L ML,TF ML and C-L-TF ML models were selected.The area under the curve(AUC)of the best ML models were compared,and the clinical value of the best TF ML model was observed with decision curve analysis(DCA).Results Totally 3 optimal C-L related features and 8 optimal TF were selected.Among the constructed C-L ML,TF ML and C-L-TF ML models,C-L-LR model,TF-LR model and C-L-TF-SVM model were the optimal ones,with AUC in training set of 0.891,0.985 and 0.965,while in test set of 0.676,0.971 and 0.948,respectively.No significant difference of AUC was found between TF-LR model and C-L-TF-SVM model in both training set and test set(both P＞0.05),which were both greater than those of C-L-LR model(all P＜0.05).TF-LR model achieved higher clinical benefits in both training set and test set.Conclusion Ultrasound TF-LR model of coronary artery could be used to effectively predict IKD in children.

Objective:To investigate the effect of exosomes released from triple-negative breast cancer cells (TNBC) on immune cells after radiotherapy.Methods:When TNBC (3 types: MDA-MB-231, MDA-MB-453, MDA-MB-468) reached 70% confluence, cells were irradiated with a dose of 8 Gy in one group and no intervention was given in the control group. The supernatants were collected at 48 h after irradiation. Subsequently, these supernatants were co-cultured with lymphocytes in peripheral blood mononuclear cells, and the uptake of exosomes by T cells was confirmed by fluorescence microscopy. Meanwhile, the expression of regulatory T cells (Treg) in the cells was detected using flow cytometry. Differences in Treg cell differentiation between two groups were compared by t-test (expressed as Treg cell positivity rate). Results:Transmission electron microscopy scanning and nanoparticle analysis showed that the extracellular vesicles extracted in the experiment were exosomes. Lymphocytes phagocytosed the exosomes and the exosomes were mainly concentrated in the cytoplasm after phagocytosis. Following the co-culture of 3 kinds of lymphocytes with exosomes from TNBC, there was an increase in Treg cell differentiation compared to control group (1.07%, 0.60%, 0.63% vs. 0.30%, P<0.01). In addition, exosomes released from TNBC further increased the differentiation of Treg cells after radiotherapy (MDA-MB-231 cells: 1.07% vs. 1.81%, P<0.01; MDA-MB-453 cells: 0.60% vs. 0.93%, P<0.05). Conclusions:In summary, exosomes released from TNBC can promote the differentiation of Treg cells. In addition, radiotherapy-induced exosomes released by TNBC further exacerbate the differentiation of Treg cell.