Objective To establish a flow cytometric assay for detecting heat shock protein A2(HSPA2)in sperm and explore the role of HSPA2 expression levels in predicting low fertilization rates in in vitro fertilization(IVF).Methods The principle of in-direct immunofluorescence(IIF)was used to fluorescently stain sperm HSPA2.After the sperm sample was permeabilized and sealed,rabbit anti-human HSPA2 antibody(primary antibody)and fluorescein isothiocyanate(FITC)labeled goat anti-rabbit IgG antibody(secondary antibody)were sequentially added as detection tubes.At the same time,a sample without primary anti-body was set up as a control tube,and the positive rates of the two tubes were measured by flow cytometer.The ratio of the posi-tive rate of the detection tube to the control tube(positive rate ratio)was calculated.The optimal number of sperm for detection and the optimal working dilutions of primary and secondary antibodies were explored using the chessboard method.Under the optimal conditions,the repeatability,linear range and reference range of the method were evaluated separately,in order to estab-lish a preliminary method for detecting sperm HSPA2 expression levels using flow cytometry.After the establishment of the method,preliminary testing was conducted on a total of 85 sperm samples from couples who underwent IVF at the Reproductive Medicine Center of Jiangmen Central Hospital in 2023.The ratio of HSPA2 positivity rates between the group with IVF success-ful(n=63)and the group with low fertilization rate(n=22)was compared,and the receiver operating characteristic(ROC)curve was used to analyze the threshold.Results The positive rate of HSPA2 in the control tube was relatively low,showing a low background signal,while the fluorescence signal of the detection tube was significantly enhanced,indicating that this method can effectively detect HSPA2.The optimal number of sperm samples for detection determined by the chessboard method was 2×106,and the optimal working dilutions for primary and secondary antibodies were 1∶300 and 1∶400,respectively.Evaluation of repeatability and linear range showed good methodological performance.Comparative analysis between the group with IVF success-ful and the group with low fertilization rate showed that the ratio of sperm HSPA2 positivity rate in the group with low fertilization rate(6.19±4.07)was lower than successful fertilization group(10.69±8.26),the difference was statistically significant(t=2.446,P<0.05).The ROC curve and Youden index showed that the best predictive power was achieved when the cutoffvalue for the ratio of positivity rate was 5.5067,with a sensitivity and a specificity of 71.4%,55.5%,respectively.Conclusion A flow cytometric method for detecting HSPA2 in sperm is successfully established.The expression level of sperm HSPA2 detected by this method suggests its predictive value for low fertilization rate in IVF,providing a basis for future clinical scientific selection of fertilization methods.

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 establish a flow cytometric assay for detecting heat shock protein A2(HSPA2)in sperm and explore the role of HSPA2 expression levels in predicting low fertilization rates in in vitro fertilization(IVF).Methods The principle of in-direct immunofluorescence(IIF)was used to fluorescently stain sperm HSPA2.After the sperm sample was permeabilized and sealed,rabbit anti-human HSPA2 antibody(primary antibody)and fluorescein isothiocyanate(FITC)labeled goat anti-rabbit IgG antibody(secondary antibody)were sequentially added as detection tubes.At the same time,a sample without primary anti-body was set up as a control tube,and the positive rates of the two tubes were measured by flow cytometer.The ratio of the posi-tive rate of the detection tube to the control tube(positive rate ratio)was calculated.The optimal number of sperm for detection and the optimal working dilutions of primary and secondary antibodies were explored using the chessboard method.Under the optimal conditions,the repeatability,linear range and reference range of the method were evaluated separately,in order to estab-lish a preliminary method for detecting sperm HSPA2 expression levels using flow cytometry.After the establishment of the method,preliminary testing was conducted on a total of 85 sperm samples from couples who underwent IVF at the Reproductive Medicine Center of Jiangmen Central Hospital in 2023.The ratio of HSPA2 positivity rates between the group with IVF success-ful(n=63)and the group with low fertilization rate(n=22)was compared,and the receiver operating characteristic(ROC)curve was used to analyze the threshold.Results The positive rate of HSPA2 in the control tube was relatively low,showing a low background signal,while the fluorescence signal of the detection tube was significantly enhanced,indicating that this method can effectively detect HSPA2.The optimal number of sperm samples for detection determined by the chessboard method was 2×106,and the optimal working dilutions for primary and secondary antibodies were 1∶300 and 1∶400,respectively.Evaluation of repeatability and linear range showed good methodological performance.Comparative analysis between the group with IVF success-ful and the group with low fertilization rate showed that the ratio of sperm HSPA2 positivity rate in the group with low fertilization rate(6.19±4.07)was lower than successful fertilization group(10.69±8.26),the difference was statistically significant(t=2.446,P<0.05).The ROC curve and Youden index showed that the best predictive power was achieved when the cutoffvalue for the ratio of positivity rate was 5.5067,with a sensitivity and a specificity of 71.4%,55.5%,respectively.Conclusion A flow cytometric method for detecting HSPA2 in sperm is successfully established.The expression level of sperm HSPA2 detected by this method suggests its predictive value for low fertilization rate in IVF,providing a basis for future clinical scientific selection of fertilization methods.

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 test the difficulty, discrimination, and reliability of the ICU Nurse Pressure Injury Prevention and Care Knowledge Questionnaire.Methods:Two researchers independently translated the ICU Nurse Pressure Injury Prevention and Care Knowledge Questionnaire and cross-checked it to form a Chinese version of the questionnaire. The Chinese version of the ICU Nurse Pressure Injury Prevention and Care Knowledge Questionnaire consists of 24 items, with correct answers scoring one point and incorrect answers scoring zero points, with a total score of 24 points. Convenience sampling was used to select ICU nurses from 14 GradeⅢ Class A hospitals in five provinces/autonomous regions and two municipalities in China for the survey between April and July 2023. The difficulty index, discrimination index, and Cronbach's α coefficient of the questionnaire were analyzed.Results:A total of 1 121 questionnaires were distributed, with 1 020 valid responses, yielding a valid response rate of 90.99%. The mean score of the 1 020 ICU nurses on the Chinese version of the ICU Nurse Pressure Injury Prevention and Care Knowledge Questionnaire was (16.10±5.58) , with a minimum score of 4.00 and a maximum score of 24.00. The Cronbach's α coefficient of the Chinese version of the ICU Nurse Pressure Injury Prevention and Care Knowledge Questionnaire was 0.91. The questionnaire's overall difficulty and discrimination indexes were 0.67 and 0.59, respectively.Conclusions:The Chinese version of the ICU Nurse Pressure Injury Prevention and Care Knowledge Questionnaire has appropriate difficulty, moderate discrimination, and strong reliability, making it a valuable tool for assessing ICU nurses' knowledge of pressure injury-related topics.

Objective:To test the difficulty, discrimination, and reliability of the ICU Nurse Pressure Injury Prevention and Care Knowledge Questionnaire.Methods:Two researchers independently translated the ICU Nurse Pressure Injury Prevention and Care Knowledge Questionnaire and cross-checked it to form a Chinese version of the questionnaire. The Chinese version of the ICU Nurse Pressure Injury Prevention and Care Knowledge Questionnaire consists of 24 items, with correct answers scoring one point and incorrect answers scoring zero points, with a total score of 24 points. Convenience sampling was used to select ICU nurses from 14 GradeⅢ Class A hospitals in five provinces/autonomous regions and two municipalities in China for the survey between April and July 2023. The difficulty index, discrimination index, and Cronbach's α coefficient of the questionnaire were analyzed.Results:A total of 1 121 questionnaires were distributed, with 1 020 valid responses, yielding a valid response rate of 90.99%. The mean score of the 1 020 ICU nurses on the Chinese version of the ICU Nurse Pressure Injury Prevention and Care Knowledge Questionnaire was (16.10±5.58) , with a minimum score of 4.00 and a maximum score of 24.00. The Cronbach's α coefficient of the Chinese version of the ICU Nurse Pressure Injury Prevention and Care Knowledge Questionnaire was 0.91. The questionnaire's overall difficulty and discrimination indexes were 0.67 and 0.59, respectively.Conclusions:The Chinese version of the ICU Nurse Pressure Injury Prevention and Care Knowledge Questionnaire has appropriate difficulty, moderate discrimination, and strong reliability, making it a valuable tool for assessing ICU nurses' knowledge of pressure injury-related topics.

Objective To investigate the correlation between qi stagnation and phlegm stasis syndrome in the young and middle-aged population and the detection rate of thyroid nodules.Methods The clinical data of those who participated in the questionnaire survey and took thyroid ultrasonography at Dongguan Hospital of Guangzhou University of Chinese Medicine from June 1 to December 1,2023 were collected.The clinical information covered age,gender,family history,body mass index(BMI),related symptoms,and ultrasonographic findings.And then the related data were statistically analyzed.Results(1)The clinical data of 196 cases were collected,of which 65 cases(33.16%)suffered from thyroid nodules,50 cases(25.51%)were differentiated as qi stagnation and phlegm stasis syndrome,53 cases(27.04%)had qi depression constitution of traditional Chinese medicine(TCM),55 cases(28.06%)had blood stasis constitution,and 48 cases(24.49%)had phlegm-dampness constitution.(2)The results of univariate analysis showed that the relevant factors for thyroid nodules included female,family history,qi stagnation and phlegm stasis syndrome,qi depression constitution,blood stasis constitution,phlegm-dampness constitution,dizziness and headache,neck stiffness,swallowing discomfort,lump on the surface of the body,dysmenorrhea and amenorrhea,tightness in the chest,distending pain in hypochondrium,depressed in spirits,emotional vulnerability,distending pain in breast,gloomy complexion,darkish lips,dark circles around the eyes,heaviness of the body,eyelid edema,and profuse sputum,and the differences were all statistically significant(P＜0.05 or P＜0.01).(3)Multivariate Logistic regression analysis was performed on the basis of univariate analysis,and the results showed that qi stagnation and phlegm stasis syndrome(OR:4.03,95%CI:1.85-8.77),phlegm-dampness constitution(OR:4.68,95%CI:2.06-10.63),and lump on the surface of the body(OR:2.97,95%CI:1.11-7.95)were the influencing factors for thyroid nodules.(4)A prediction model for detecting thyroid nodules was constructed:logit(P)=-1.607+1.39×qi stagnation and phlegm stasis syndrome(0 expressing absence,1 expressing presence)+1.54×phlegm-dampness constitution(0 expressing absence,1 expressing presence)+1.09×lump on the surface of the body(0 expressing absence,1 expressing presence).The model was evaluated by using the receiver operating characteristic(ROC)curve,and the area under the curve(AUC)was 0.75(95%CI:0.67-0.83,P＜0.001).Conclusion In the young and middle-aged population,qi stagnation and phlegm stasis are the risk factors for the detectable rate of thyroid nodules.The early identification,risk prediction and timely intervention for the population with qi stagnation and phlegm stasis will be helpful for the prevention and treatment of thyroid nodules.