This study aims to establish a high-performance size-exclusion chromatography (HPSEC) method for determining the content of the classical swine fever virus (CSFV) E2 protein and screen the optimal stabilizer to enhance the stability of this protein. The optimal detection conditions were determined by optimizing the composition of the mobile phase, and characteristic chromatographic peaks were identified by SDS-PAGE and Western blotting. The specificity, repeatability, precision, linearity, limit of detection (LOD), and limit of quantitation (LOQ) of the method were assessed. The method established was used to determine the content of CSFV E2 protein antigen and vaccine. Differential scanning fluorimetry (DSF) was employed to screen the buffer system, pH, and salt ion concentrations, and sugar, amino acid, and alcohol stabilizers were further screened. The results showed that using a 200 mmol/L phosphate buffer provided the best column efficiency. An antigen-specific chromatographic peak appeared at the retention time of 18 min, which was identified as the CSFV E2 protein by SDS-PAGE and Western blotting. The method exhibited high specificity for detecting the CSFV E2 protein, with no absorbance peak observed in the blank control. The relative standard deviation (RSD) of the peak area for six repeated injections of the CSFV E2 protein was 0.74%, indicating good repeatability of the method. The RSD for repeated detection of two different concentrations of CSFV E2 protein samples by different operators at different time points was less than 2%, suggesting good intermediate precision of the method. The peak area of the CSFV E2 protein was linearly related to its concentration, with the regression equation showing R² of 1.000. The LOD and LOQ of the method were 14.88 μg/mL and 29.75 μg/mL, respectively. Application of the developed method in the detection of three batches of CSFV E2 protein antigen and three batches of vaccine demonstrated results consistent with those from the bicinchoninic acid (BCA) assay, which meant that the method could accurately determine the content of CSFV E2 protein antigen and vaccine. The DSF method identified 50 mmol/L Tris-HCl at pH 8.0 as the optimal buffer, and the addition of sugar and alcohol stabilizers further improved the stability of the CSFV E2 protein. The HPSEC method established in this study is simple, fast, and exhibits good accuracy and repeatability, enabling precise measurement of the CSFV E2 protein content. It is expected to play a crucial role in the quality control of the CSFV E2 vaccine. Furthermore, the strategy for improving the CSFV E2 protein stability, identified through DSF screening, has significant implications for enhancing the stability of the CSFV E2 vaccine.
Classical Swine Fever Virus/chemistry* ; Chromatography, Gel/methods* ; Animals ; Swine ; Viral Envelope Proteins/immunology*

Objective:To explore the application value of echocardiography in the management of pediatric mitral valve replacement.Methods:Thirty-three children who underwent mitral valve replacement at Union Hospital Tongji Medical College Huazhong University of Science and Technology from January 2009 to June 2023 were retrospectively analyzed. Clinical data and preoperative and postoperative echocardiographic data were collected. The differences in ultrasound parameters among preoperative，1-week postoperative，1-month postoperative and 1-year postoperative were compared.Results:Of the 33 children，there were 4 cases（12.12%）of mitral stenosis，33 cases（100%）of mitral regurgitation and 4 cases（12.12%）of mitral stenosis combined with regurgitation. Mechanical valve replacement was performed in 30 cases（90.91%）and bioprosthetic valve replacement in 3 cases（9.09%）. Compared with the preoperative period，the left atrial internal diameter and left ventricular end-diastolic internal diameter were significantly reduced at 1-week postoperatively，1-month postoperatively，and 1-year postoperatively，respectively（all P <0.05）.The left ventricular mass was significantly reduced and the left ventricular end-diastolic sphericity index was significantly increased（both P < 0.05）at 1-month postoperatively compared with preoperatively. The left ventricular ejection fraction was significantly higher at 1-year postoperatively compared with 1-week postoperatively（ P <0.05）. Conclusions:After pediatric mitral valve replacement，the left ventricular dimensions decrease，and cardiac function progressively recovers. Echocardiography is an essential tool for accurate preoperative assessment and postoperative follow-up in pediatric mitral valve replacement.

Objective:To investigate the correlation between frailty and telomere length in the elderly.Methods:Cross-sectional study.A total of 128 elderly patients aged ≥65 years who were hospitalized in the Second Hospital of Shanxi Medical University from January to June 2024 were selected.Frailty assessment was conducted using the Edmonton Scale, and patients were divided into the non-frail group(n=64)and the frail group(n=64)based on frailty score.The telomere length of peripheral blood leukocyte was detected by real-time fluorescent quantitative polymerase chain reaction(qPCR), and the correlation analysis between frailty and telomere length was conducted.Results:Compared with the non-frailty group, the frailty group had older age, higher body mass index(BMI), higher American Society of Anesthesiologists(ASA)grade, an increased proportion of diabetes and hypertension, and shorter telomere length( P<0.05). Logistic regression analysis: ageing, increased BMI, hypertension and diabetes were risk factors for frailty in the elderly( OR=1.280, 1.135, 1.543, 1.081, P=0.001, 0.036, 0.010, 0.021), while ASA grade and telomere length were not related to the occurrence of frailty.After stratification, frailty scores were weakly correlated with telomere length in the age ≤73 years old ( r=-0.344, P=0.005)and the BMI ≥24 kg/m 2( r=-0.336, P=0.001). The correlation between frailty scores and telomere length was strengthened in the poorly controlled hypertension group( r=-0.571, P=0.042)and the group with poorly controlled diabetes( r=-0.613, P=0.045)showed a stronger correlation between frailty scores and telomere length. Conclusions:In the overall analysis, there was no definitive association between telomere length and frailty.After stratification, it was found that telomere length was weakly to moderately correlated with frailty, suggesting that telomere length may interact with other factors, and strong risk factors such as age, BMI, hypertension and diabetes may mask the weak effect of telomere length on frailty.

Objective:To investigate the correlation between frailty and telomere length in the elderly.Methods:Cross-sectional study.A total of 128 elderly patients aged ≥65 years who were hospitalized in the Second Hospital of Shanxi Medical University from January to June 2024 were selected.Frailty assessment was conducted using the Edmonton Scale, and patients were divided into the non-frail group(n=64)and the frail group(n=64)based on frailty score.The telomere length of peripheral blood leukocyte was detected by real-time fluorescent quantitative polymerase chain reaction(qPCR), and the correlation analysis between frailty and telomere length was conducted.Results:Compared with the non-frailty group, the frailty group had older age, higher body mass index(BMI), higher American Society of Anesthesiologists(ASA)grade, an increased proportion of diabetes and hypertension, and shorter telomere length( P<0.05). Logistic regression analysis: ageing, increased BMI, hypertension and diabetes were risk factors for frailty in the elderly( OR=1.280, 1.135, 1.543, 1.081, P=0.001, 0.036, 0.010, 0.021), while ASA grade and telomere length were not related to the occurrence of frailty.After stratification, frailty scores were weakly correlated with telomere length in the age ≤73 years old ( r=-0.344, P=0.005)and the BMI ≥24 kg/m 2( r=-0.336, P=0.001). The correlation between frailty scores and telomere length was strengthened in the poorly controlled hypertension group( r=-0.571, P=0.042)and the group with poorly controlled diabetes( r=-0.613, P=0.045)showed a stronger correlation between frailty scores and telomere length. Conclusions:In the overall analysis, there was no definitive association between telomere length and frailty.After stratification, it was found that telomere length was weakly to moderately correlated with frailty, suggesting that telomere length may interact with other factors, and strong risk factors such as age, BMI, hypertension and diabetes may mask the weak effect of telomere length on frailty.

Objective:To explore the application value of echocardiography in the management of pediatric mitral valve replacement.Methods:Thirty-three children who underwent mitral valve replacement at Union Hospital Tongji Medical College Huazhong University of Science and Technology from January 2009 to June 2023 were retrospectively analyzed. Clinical data and preoperative and postoperative echocardiographic data were collected. The differences in ultrasound parameters among preoperative，1-week postoperative，1-month postoperative and 1-year postoperative were compared.Results:Of the 33 children，there were 4 cases（12.12%）of mitral stenosis，33 cases（100%）of mitral regurgitation and 4 cases（12.12%）of mitral stenosis combined with regurgitation. Mechanical valve replacement was performed in 30 cases（90.91%）and bioprosthetic valve replacement in 3 cases（9.09%）. Compared with the preoperative period，the left atrial internal diameter and left ventricular end-diastolic internal diameter were significantly reduced at 1-week postoperatively，1-month postoperatively，and 1-year postoperatively，respectively（all P <0.05）.The left ventricular mass was significantly reduced and the left ventricular end-diastolic sphericity index was significantly increased（both P < 0.05）at 1-month postoperatively compared with preoperatively. The left ventricular ejection fraction was significantly higher at 1-year postoperatively compared with 1-week postoperatively（ P <0.05）. Conclusions:After pediatric mitral valve replacement，the left ventricular dimensions decrease，and cardiac function progressively recovers. Echocardiography is an essential tool for accurate preoperative assessment and postoperative follow-up in pediatric mitral valve replacement.

Purpose/Significance The processing links and key technologies of health care big data are deeply discussed to provide in-formation support and guarantee for further promoting the application of health care big data.Method/Process According to the scientific research paradigm driven by big data,and based on the process perspective,the paper introduces the sources and characteristics of health care big data,analyzes the processing links and key technologies of big data,and discusses the current challenges and future development directions of health care big data processing.Result/Conclusion Both China and foreign countries have achieved innovative development in technologies for health care big data processing.However,there are still some challenges in multi-dimensional data collection,multi-mo-dal data integration,large model data analysis,and data security.In the future,through building a comprehensive platform of big data pro-cessing,and developing or integrating targeted functional modules,the existing problems can be effectively addressed.

Objective:To evaluate the diagnostic efficacy and practicality of the Jaundice color card (JCard) as a screening tool for neonatal jaundice.Methods:Following the standards for reporting of diagnostic accuracy studies (STARD) statement, a multicenter prospective study was conducted in 9 hospitals in China from October 2019 to September 2021. A total of 845 newborns who were admitted to the hospital or outpatient department for liver function testing due to their own diseases. The inclusion criteria were a gestational age of ≥35 weeks, a birth weight of ≥2 000 g, and an age of ≤28 days. The neonate′s parents used the JCard to measure jaundice at the neonate′s cheek. Within 2 hours of the JCard measurement, transcutaneous bilirubin (TcB) was measured with a JH20-1B device and total serum bilirubin (TSB) was detected. The Pearson′s correlation analysis, Bland-Altman plots and the receiver operating characteristic (ROC) curve were used for statistic analysis.Results:Out of the 854 newborns, 445 were male and 409 were female; 46 were born at 35-36 weeks of gestational age and 808 were born at ≥37 weeks of gestational age. Additionally, 432 cases were aged 0-3 days, 236 cases were aged 4-7 days, and 186 cases were aged 8-28 days. The TSB level was (227.4±89.6) μmol/L, with a range of 23.7-717.0 μmol/L. The JCard level was (221.4±77.0) μmol/L and the TcB level was (252.5±76.0) μmol/L. Both the JCard and TcB values showed good correlation ( r=0.77 and 0.80, respectively) and agreements (96.0% (820/854) and 95.2% (813/854) of samples fell within the 95% limits of agreement, respectively) with TSB. The JCard value of 12 had a sensitivity of 0.93 and specificity of 0.75 for identifying a TSB ≥205.2?μmol/L, and a sensitivity of 1.00 and specificity of 0.35 for identifying a TSB ≥342.0?μmol/L. The TcB value of 205.2?μmol/L had a sensitivity of 0.97 and specificity of 0.60 for identifying TSB levels of 205.2 μmol/L, and a sensitivity of 1.00 and specificity of 0.26 for identifying TSB levels of 342.0 μmol/L. The areas under the ROC curve (AUC) of JCard for identifying TSB levels of 153.9, 205.2, 256.5, and 342.0 μmol/L were 0.96, 0.92, 0.83, and 0.83, respectively. The AUC of TcB were 0.94, 0.91, 0.86, and 0.87, respectively. There were both no significant differences between the AUC of JCard and TcB in identifying TSB levels of 153.9 and 205.2 μmol/L (both P>0.05). However, the AUC of JCard were both lower than those of TcB in identifying TSB levels of 256.5 and 342.0 μmol/L (both P<0.05). Conclusions:JCard can be used to classify different levels of bilirubin, but its diagnostic efficacy decreases with increasing bilirubin levels. When TSB level are ≤205.2 μmol/L, its diagnostic efficacy is equivalent to that of the JH20-1B. To prevent the misdiagnosis of severe jaundice, it is recommended that parents use a low JCard score, such as 12, to identify severe hyperbilirubinemia (TSB ≥342.0 μmol/L).

Approximately 140 million people worldwide are homozygous carriers of APOE4 (ε4), a strong genetic risk factor for late onset familial and sporadic Alzheimer's disease (AD), 91% of whom will develop AD at earlier age than heterozygous carriers and noncarriers. Susceptibility to AD could be reduced by targeted editing of APOE4, but a technical basis for controlling the off-target effects of base editors is necessary to develop low-risk personalized gene therapies. Here, we first screened eight cytosine base editor variants at four injection stages (from 1- to 8-cell stage), and found that FNLS-YE1 variant in 8-cell embryos achieved the comparable base conversion rate (up to 100%) with the lowest bystander effects. In particular, 80% of AD-susceptible ε4 allele copies were converted to the AD-neutral ε3 allele in human ε4-carrying embryos. Stringent control measures combined with targeted deep sequencing, whole genome sequencing, and RNA sequencing showed no DNA or RNA off-target events in FNLS-YE1-treated human embryos or their derived stem cells. Furthermore, base editing with FNLS-YE1 showed no effects on embryo development to the blastocyst stage. Finally, we also demonstrated FNLS-YE1 could introduce known protective variants in human embryos to potentially reduce human susceptivity to systemic lupus erythematosus and familial hypercholesterolemia. Our study therefore suggests that base editing with FNLS-YE1 can efficiently and safely introduce known preventive variants in 8-cell human embryos, a potential approach for reducing human susceptibility to AD or other genetic diseases.
Humans ; Apolipoprotein E4/genetics* ; Cytosine ; Mutation ; Blastocyst ; Heterozygote ; Gene Editing ; CRISPR-Cas Systems

Objective To evaluate the role of a color jaundice card (6 colors) as a possible screening tool for detecting neonatal hyperbilirubinemia.Methods During February 1,2016 and May 31,2017,neonates were enrolled in the study,with gestational age ≥35 weeks,birth weight ≥2 000 g,postnatal age 3-28 days,who were the outpatients or inpatients of the 9th People's Hospital of Wuxi Affiliated to Soochow University and the People's Hospital of Anyang.In a well-lighted room,the card measurements were performed at the infants' forehead,the cheek and the sternum.The skin was pressed with a finger for 2 seconds and left quickly,and then the card was used to compare with the exposed yellow skin.Within 2 hours after jaundice card measurement,blood was obtained by venipuncture and total serum bilirubin (TSB) levels were measured.The sensitivity,specificity,positive predictive value (PPV),negative predictive value (NPV),positive likelihood ratio (PLR) and negative likelihood ratio (NLR) were calculated at each measurement sites.Results One hundred and thirty-two neonates were enrolled,of whom 68 cases (51.5％) were male and 64 cases(48.5％) were female and 18 cases (13.6％) were preterm and 114 cases (86.4％) were term neonates.Among all neonates,TSB was ＜5.00 mg/dL(1 mg/dL =17.1 μmol/L) in 21 cases (15.9％),5.00-9.99 mg/dL in 26 cases (19.7％),10.00-14.99 mg/dL in 34 cases (25.8％),15.00-19.99 mg/dL in 37 cases (28.0％) and ≥ 20.00 mg/dL in 14 cases (10.6％).The card had the highest cap ability to recognize jaundice at the cheek,slightly lower at the sternum and the worst in the forehead.The cut-off of ≥ 12 on the six-color card at the cheek had a sensitivity of 95.95％,specificity of 74.14％,PPV of 82.56％,NPV of 93.48％,PLR of 3.710 and NLR of 0.055 for identifying neonates with TSB ≥ 12 mg/dL,with sensitivity being 98.08％,specificity 57.50％,PPV 60.00％,NPV 97.87％,PLR 2.308 and NLR 0.033 for TSB≥ 15 mg/dL.The identification rate was as follows:sensitivity of 100.00％,specificity of 46.00％,PPV of 37.21％,NPV of 100.00％ and PLR of 1.852 for predicting TSB ≥ 17 mg/dL.In addition,in the forehead,cheeks and sternum,the sensitivity of the cut-off of ≥ 12 on the card was 100.00％ for identifying neonates with TSB≥20 mg/dL.In the cheeks and the sternum,the cut-off of ≥ 15 on the card was with a sensitivity of 100.00％ for predicting TSB ≥ 20 mg/dL.Conclusion The six-color jaundice card is a potential screening tool for neonatal hyperbilirubinemia,and the cheek is the best measurement site.

Objective To observe the effects of live trigeminal bifidobacterium, lactobacillus and enterococcus powder on the recurrence of wheezing, and the levels of peripheral blood eosinophil ( EOS) and serum transforming growth factor-beta 1( TGF-β1) in atopic children with bronchiolitis.Methods Sixty atopic children with bronchiolitis were randomly divided into the therapy group (30 cases) and the conventional treatment group (30 cases) and another 25 healthy children were recruited as the healthy control group. The conventional treatment group was given routine therapy, and the therapy group received live trigeminal bifidobacterium, lactobacil-lus and enterococcus, in addition to routine therapy for 2 months.The levels of EOS and TGF-β1 were detected at the acute stage and 2 months after receiving trigeminal bifidobacterium, lactobacillus and enterococcus.Results (1) The recurrent rate of wheezing after medication for the therapy group (0.67 ±0.13) was significantly lower than that for the conventional treatment group (1.27 ±0.17), with statistical significance (P<0.05).(2)The levels of EOS of the therapy group [(0.72 ±0.13) ×109/L] and the conventional treatment group [(0.70 ±0.13) ×109/L] at the acute stage were markedly higher than those of the healthy control group [(0.16 ± 0.09) ×109/L], also with statistical significance (P<0.05).The levels of TGF-β1 of the therapy group [(1.20 ±0.13) ng/L] and the conventional treatment group(1.22 ±0.11) at acute stage were all considerably lower than those of the control group [(1.45 ± 0.13) ng/L], with statistical significance (P<0.05).The level of EOS in the therapy group [(0.27 ±0.12) ×109/L] 2 months af-ter medication of oral live trigeminal bifidobacterium, lactobacillus and enterococcus powder was lower than that in the conventional treatment group [(0.36 ±0.14) ×109/L], also with statistical significance (P<0.05).The level of TGF-β1 of the therapy group [(1.41 ±0.09) ng/L] 2 months after medication was markedly higher than that of the conventional treatment group [(1.34 ±0.10) ng/L], also with statistical significance (P<0.05).Conclusion Oral medication of live trigeminal bifidobacterium, lactobacillus and enterococcus powder for 2 months could obviously reduce the recurrent rate of wheezing within 6 months after the onset of bronchiolitis and could also up-regulate the levels of EOS and TGF-β1 in atopic children with bronchiolitis.