The amino-terminal pro-C-type natriuretic peptide(NT-proCNP)is a diagnostic inflam-matory marker clinically used for diagnosing bacterial infections.This study aims to establish a phage display library of single-chain variable fragment(scFv)antibodies against canine NT-proC-NP and to screen for scFvs with high binding affinity to NT-proCNP.Initially,NT-proCNP was prepared using prokaryotic expression system and was used to immunize New Zealand White rab-bits.Upon achieving the desired serum titer,total RNA was extracted from the splenocytes of rab-bits and reverse transcribed into cDNA.Using this cDNA as a template,degenerate primers were employed to amplify the genes of the rabbit antibody light chain variable region(VL)and heavy chain variable region(VH).The VL and VH regions were spliced together to form a complete scFv fragment via overlap extension PCR.The scFv was then ligated into the phagemid pComb3XSS and electroporated into competent E.coli TG1 cells to construct a rabbit-derived anti-NT-proCNP scFv immunological library.This library underwent four rounds of enrichment and screening to isolate specific single-chain antibodies.The selected antibody was subsequently ex-pressed in a soluble form within a prokaryotic system,and its immunological activity was evalua-ted.Using phage display technology,this study successfully identified a single-chain antibody scFv-1-CNP with strong antigen-binding activity and genetic sequence characteristics of scFvs,providing a research direction for further exploration of scFv applications in the detection of NT-proCNP.

The amino-terminal pro-C-type natriuretic peptide(NT-proCNP)is a diagnostic inflam-matory marker clinically used for diagnosing bacterial infections.This study aims to establish a phage display library of single-chain variable fragment(scFv)antibodies against canine NT-proC-NP and to screen for scFvs with high binding affinity to NT-proCNP.Initially,NT-proCNP was prepared using prokaryotic expression system and was used to immunize New Zealand White rab-bits.Upon achieving the desired serum titer,total RNA was extracted from the splenocytes of rab-bits and reverse transcribed into cDNA.Using this cDNA as a template,degenerate primers were employed to amplify the genes of the rabbit antibody light chain variable region(VL)and heavy chain variable region(VH).The VL and VH regions were spliced together to form a complete scFv fragment via overlap extension PCR.The scFv was then ligated into the phagemid pComb3XSS and electroporated into competent E.coli TG1 cells to construct a rabbit-derived anti-NT-proCNP scFv immunological library.This library underwent four rounds of enrichment and screening to isolate specific single-chain antibodies.The selected antibody was subsequently ex-pressed in a soluble form within a prokaryotic system,and its immunological activity was evalua-ted.Using phage display technology,this study successfully identified a single-chain antibody scFv-1-CNP with strong antigen-binding activity and genetic sequence characteristics of scFvs,providing a research direction for further exploration of scFv applications in the detection of NT-proCNP.

Background::Carotid intima-media thickness (IMT) and diameter, stiffness, and wave reflections, are independent and important clinical biomarkers and risk predictors for cardiovascular diseases. The purpose of the present study was to establish nationwide reference values of carotid properties for healthy Chinese adults and to explore potential clinical determinants.Methods::A total of 3053 healthy Han Chinese adults (1922 women) aged 18-79 years were enrolled at 28 collaborating tertiary centers throughout China between April 2021 and July 2022. The real-time tracking of common carotid artery walls was achieved by the radio frequency (RF) ultrasound system. The IMT, diameter, compliance coefficient, β stiffness, local pulse wave velocity (PWV), local systolic blood pressure, augmented pressure (AP), and augmentation index (AIx) were then automatically measured and reported. Data were stratified by age groups and sex. The relationships between age and carotid property parameters were analyzed by Jonckheere-Terpstra test and simple linear regressions. The major clinical determinants of carotid properties were identified by Pearson’s correlation, multiple linear regression, and analyses of covariance.Results::All the parameters of carotid properties demonstrated significantly age-related trajectories. Women showed thinner IMT, smaller carotid diameter, larger AP, and AIx than men. The β stiffness and PWV were significantly higher in men than women before forties, but the differences reversed after that. The increase rate of carotid IMT (5.5 μm/year in women and 5.8 μm/year in men) and diameter (0.03 mm/year in both men and women) were similar between men and women. For the stiffness and wave reflections, women showed significantly larger age-related variations than men as demonstrated by steeper regression slopes (all P for age by sex interaction <0.05). The blood pressures, body mass index (BMI), and triglyceride levels were identified as major clinical determinants of carotid properties with adjustment of age and sex. Conclusions::The age- and sex-specific reference values of carotid properties measured by RF ultrasound for healthy Chinese adults were established. The blood pressures, BMI, and triglyceride levels should be considered for clinical application of corresponding reference values.

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).

Purpose To explore the value of contrast-enhanced ultrasound(CEUS)in the differential diagnosis of gallbladder polypoid lesions(GPLs)(diameter≥10 mm).Materials and Methods A prospective enrollment of 229 patients with GPLs who underwent cholecystectomy in 17 hospitals from December 1 2021 to June 30 2024 was conducted to analyze the relationship between general data,conventional ultrasound,CEUS characteristics and the nature of GPLs.Multivariate Logistic regression was employed to identify independent risk factors for neoplastic polyps,the differential diagnostic value of different indicators was compared.Results Among 229 patients with GPLs,there were 108 cases of cholesterol polyps,102 cases of adenoma and 19 cases of gallbladder cancer.Age(Z=-4.476,P<0.001),polyp number(χ2=15.561,P<0.001),diameter(Z=-8.149,P<0.001),echogenicity(χ2=9.241,P=0.010),vascularity(χ2=23.107,P<0.001),enhancement intensity(χ2=47.610,P<0.001),enhancement pattern(χ2=6.468,P=0.011),vascular type(χ2=84.470,P<0.001),integrity of gallbladder wall(χ2=7.662,P=0.006)and stalk width(Z=-9.831,P<0.001)between cholesterol polyps and neoplastic polyps were statistically significant.Age,location,diameter,echogenicity,enhancement pattern,vascular type and stalk width between adenoma and gallbladder cancer were statistically significant(Z=-4.333,-3.902,-5.042,all P<0.05).Multivariate Logistic regression analysis showed that hyper-enhancement,branched vascular type and stalk width were independent risk factors for neoplastic polyps(OR=4.563,5.770,3.075,all P<0.001).The combination of independent risk factors was better than single factor and diameter in the differential diagnosis of cholesterol polyps and neoplastic polyps(all P<0.01).Conclusion CEUS can effectively identify the nature of GPLs and provide a valuable imaging reference for the selection of treatment methods.

Objective To prepare a kind of lipid nanoparticle ultrasound contrast agents with the ability to target to viable myocardium for diagnosis . Methods T he agent was a biotinylated ,fluorescent‐labelled ,lipid‐coated , liquid perfluorocarbon emulsion . Physico‐chemical properties of the agent were measured ,including size distribution ,Zeta Potential ,concentration and so on . Ischemia‐reperfusion models were created in rats ,and then exposed to biotinylated anti‐MCP‐1 monoclonal antibody ,rhodamine avidin and biotinylated ,FITC‐labelled nanoparticles ,respectively . Echocardiography was taken before and after injection . Frozen sections of their hearts were observed under fluorescence microscope . Results T he particle diameter ,zeta potential and concentration of lipid nanoparticles were ( 172 .30 ± 52 .06) nm ,( －33 .10 ± 6 .50) mV and ( 2 .28 ± 0 .46 ) × 1011/ml ,respectively . From the short‐axis view ,the myocardium under endocardium of anterior wall was enhanced obviously . While myocardium of other walls were still . T he lipid nanoparticles located in the myocardium of anterior wall and gave out bright green and red fluorescence under fluorescence microscope ,w hile neither lipid nanoparticles nor fluorescence were found in other sites of ventricular myocardium . Conclusions The viable myocardium can be targeted and acoustically enhanced by the self‐made nano‐scale ultrasound contrast agent . T his new agent has potential to improve sensitivity and specificity for noninvasive identifying viable myocardium .

Objective: To prepare a kind of lipid nanoparticle ultrasound contrast agents with the ability to target to viable myocardium for diagnosis. Methods: The agent was a biotinylated, fluorescent-labelled, lipid-coated, liquid perfluorocarbon emulsion. Physico-chemical properties of the agent were measured, including size distribution, Zeta Potential, concentration and so on. Ischemia-reperfusion models were created in rats, and then exposed to biotinylated anti-MCP-1 monoclonal antibody, rhodamine avidin and biotinylated, FITC-labelled nanoparticles, respectively. Echocardiography was taken before and after injection. Frozen sections of their hearts were observed under fluorescence microscope. Results: The particle diameter, zeta potential and concentration of lipid nanoparticles were (172.30±52.06)nm, (-33.10±6.50)mV and (2.28±0.46)×10¹¹/ml, respectively. From the short-axis view, the myocardium under endocardium of anterior wall was enhanced obviously. While myocardium of other walls were still. The lipid nanoparticles located in the myocardium of anterior wall and gave out bright green and red fluorescence under fluorescence microscope, while neither lipid nanoparticles nor fluorescence were found in other sites of ventricular myocardium. Conclusions The viable myocardium can be targeted and acoustically enhanced by the self-made nano-scale ultrasound contrast agent. This new agent has potential to improve sensitivity and specificity for noninvasive identifying viable myocardium.

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.

To explore the diagnostic value of copeptin (CPP) in cardiorenal syndrome (CRS) in rats and the association between CPP and impairment of heart and kidney, 60 male SD rats were randomly divided into blank control group (CK group), kidney failure group (SNX group), heart failure group (MI group), and CRS group. Heart and kidney function and their histology changes in rats from each group were detected. The correlation between serum CPP and heart and kidney function indexes was performed with Pearson correlation analysis. The HE staining of heart and kidney showed that the tissue lesion was more severe in CRS group than in SNX group and MI group. There was a significant positive correlation between serum CPP and brain natriuretic peptide (BNP) (r=0.638, P<0.05). No correlation was observed between serum CPP and cardiac function index (left ventricular systolic pressure, left ventricular diastolic pressure, left ventricular end-diastolic pressure) or renal function index (serum creatinine, urine creatinine, blood urea nitrogen) (r=0.512, 0.189, -0.063, 0.207, 0.290, 0.595, respectively, all P>0.05). The CPP level is associated with the degree of heart and kidney damage in CRS rats.

Objective To predict the value of advanced glycation end products (AGEs) in the risk and severity of coronary heart disease in diabetic patients.Methods Totally 120 cases were divided into 3 groups.Group A had no diabetes mellitus (DM) and no coronary atherosclerotic heart disease (CAD).Group B had DM without CAD.Group C had DM with CAD.The levels of AGEs,low density lipoprotein cholesterol (LDL-C),glycosylated hemoglobin,and the severity of coronary stenosis were detected.Receiver operating characteristic (ROC) curve was used to evaluate the sensitivities and specificities of AGEs for the diagnosis of DM with CAD.Results The highest level of AGEs,LDL-C,glycosylated hemoglobin and carotid plaque was in the C group,with a statistically significant difference (P ＜ 0.05).The severity of AGEs was significantly correlated with the severity of CAD and the Gensini score of AGEs (r =0.445).ROC curve showed that the sensitivity,specificity and area under ROC curve of AGEs determination of DM with CAD were 80.0％,75.0％,and 0.86,respectivity.Conclusions The level of AGEs is significantly correlated with atherosclerosis and prognosis in DM.The severity of CAD was higher in the patients with higher AGEs,and the incidence and severity of restenosis increased.