1.Establishment of single-chain antibody library targeting canine NT-proCNP,and screening and immune activity detection of a selected single-chain antibody
Shaojia JIANG ; Sha NAN ; Huikang WANG ; Ling MAO ; Ruiling YIN ; Qianghui LEI ; Haolong WANG ; Hao LI ; Jinyu XIAO ; Mingxing DING ; Yi DING
Chinese Journal of Veterinary Science 2025;45(3):535-541
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.
2.Applications and mechanisms of the therapeutic low-intensity pulsed ultrasound in the immune cell modulation
Yingxin LI ; Tianshu LIU ; Qiaofeng JIN ; Xueke LI ; Jiani QIU ; Jingrong JIANG ; Mingxing XIE ; Jing WANG
Chinese Journal of Ultrasonography 2025;34(3):270-276
Low-intensity pulsed ultrasound(LIPUS),with its remarkable advantages of higher safety and better penetrability,has gradually become a novel method of physical adjuvant therapy. Previous studies have verified that LIPUS can modulate the immune response of different immune cells such as macrophage,T lymphocyte,and neutrophil by reducing the level of pro-inflammatory cytokines. Therefore,it plays a crucial role on acceleration of fracture healing,expedition of wound repair,and repairation of myocardial injury. The review summarizes the regulatory effects and potential mechanisms of LIPUS on abnormal immune cell responses triggered by various diseases.
3.Establishment of single-chain antibody library targeting canine NT-proCNP,and screening and immune activity detection of a selected single-chain antibody
Shaojia JIANG ; Sha NAN ; Huikang WANG ; Ling MAO ; Ruiling YIN ; Qianghui LEI ; Haolong WANG ; Hao LI ; Jinyu XIAO ; Mingxing DING ; Yi DING
Chinese Journal of Veterinary Science 2025;45(3):535-541
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.
4.Applications and mechanisms of the therapeutic low-intensity pulsed ultrasound in the immune cell modulation
Yingxin LI ; Tianshu LIU ; Qiaofeng JIN ; Xueke LI ; Jiani QIU ; Jingrong JIANG ; Mingxing XIE ; Jing WANG
Chinese Journal of Ultrasonography 2025;34(3):270-276
Low-intensity pulsed ultrasound(LIPUS),with its remarkable advantages of higher safety and better penetrability,has gradually become a novel method of physical adjuvant therapy. Previous studies have verified that LIPUS can modulate the immune response of different immune cells such as macrophage,T lymphocyte,and neutrophil by reducing the level of pro-inflammatory cytokines. Therefore,it plays a crucial role on acceleration of fracture healing,expedition of wound repair,and repairation of myocardial injury. The review summarizes the regulatory effects and potential mechanisms of LIPUS on abnormal immune cell responses triggered by various diseases.
5.Contrast-Enhanced Ultrasound in the Differential Diagnosis of Gallbladder Polypoid Lesions:A Multicenter Study
Ligang JIA ; Xiang FEI ; Xiang JING ; Mingxing LI ; Fang NIE ; Dong JIANG ; Shaoshan TANG ; Wei ZHANG ; Hong DING ; Tao SONG ; Qi ZHOU ; Bei ZHANG ; Zhixia SUN ; Xiaojuan MA ; Nianan HE ; Fang LI ; Yingqiao ZHU ; Wen CHENG ; Yukun LUO
Chinese Journal of Medical Imaging 2024;32(11):1147-1154
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.
6.Effectiveness of 5G-based robot-assisted remote thoracolumbar internal fixation surgery
Da HE ; Qi ZHANG ; Jingwei ZHAO ; Mingxing FAN ; Bo LIU ; Xieyuan JIANG
Chinese Journal of Orthopaedics 2024;44(15):995-1001
Objective:To assess the effectiveness and viability of 5G-enabled robot-assisted thoracolumbar internal fixation surgery, scrutinizing its benefits and potential limitations in a clinical setting.Methods:This retrospective case series study analyzed 61 patients who underwent robot-assisted remote thoracolumbar internal fixation surgery between April and December 2021. The orthopaedic robotic telesurgery system, centered at Beijing Jishuitan Hospital and utilizing 5G technology for network transmission, was used to assist in screw placement. The surgeries, distributed across ten hospitals and executed over an average distance of 1 617 km (maximum exceeding 3 000 km), were appraised by examining the accuracy of screw placement, intraoperative communication efficacy, complication rates, among other metrics, and benchmarked against traditional local procedures.Results:Telesurgery was completed successfully in all patients, averaging 92±32 minutes per operation. Out of 354 screws placed, 89.8% were positioned perfectly and 97.5% were deemed clinically acceptable. The 61 patients, followed-up for an average of 18.9±1.9 months, exhibited significant postoperative improvement in pain. The preoperative visual analogue scale (VAS) pain score was 5.74±1.30 and improved to 2.30±0.90 at discharge, a statistically significant difference ( t=11.002, P<0.001). 6 patients with thoracolumbar degenerative spine disease showed significant relief of claudication and pain symptoms postoperatively. 55 patients with thoracolumbar spine fractures had a preoperative height ratio of 62.78%±12.60% at the anterior margin of the injured vertebrae, which recovered to 85.36%±8.35% postoperatively, a statistically significant difference ( t=16.977, P<0.001). All fractures were healed at final follow-up. No perioperative complications, such as vascular or nerve injuries or infections, occurred. Additionally, there were no complications related to the use of the robot. The 5G network ensured stable transmission of robotic control signals, images, and audio signals during the procedure, with no signal delays or communication failures observed. The perfect rate (90.2% vs. 93.8%, χ 2=0.169, P=0.681) and clinical acceptability rate (99.0% vs. 100.0%, P=1.000) of remote versus local robotic-assisted screw placement in participating hospital 1 (Shantou Xin Sheng Orthopaedic Hospital) were similar. The perfect rate (87.0% vs. 85.4%, χ 2=0.075, P=0.784) and clinical acceptability rate (100.0% vs. 96.6%, χ 2=0.580, P=0.446) of remote versus local robotic-assisted screw placement in participating hospital 2 (Guizhou Provincial People's Hospital) were similar. Conclusion:5G technology enhances the remote application of robotic thoracolumbar surgery without compromising patient safety or outcome quality. It underscores the potential for broader adoption of surgical robotics, aiding in the standardization of medical practices across China, boosting healthcare efficiency, and aligning with the Healthy China strategy.
7.Multicenter evaluation of the diagnostic efficacy of jaundice color card for neonatal hyperbilirubinemia
Guochang XUE ; Huali ZHANG ; Xuexing DING ; Fu XIONG ; Yanhong LIU ; Hui PENG ; Changlin WANG ; Yi ZHAO ; Huili YAN ; Mingxing REN ; Chaoying MA ; Hanming LU ; Yanli LI ; Ruifeng MENG ; Lingjun XIE ; Na CHEN ; Xiufang CHENG ; Jiaojiao WANG ; Xiaohong XIN ; Ruifen WANG ; Qi JIANG ; Yong ZHANG ; Guijuan LIANG ; Yuanzheng LI ; Jianing KANG ; Huimin ZHANG ; Yinying ZHANG ; Yuan YUAN ; Yawen LI ; Yinglin SU ; Junping LIU ; Shengjie DUAN ; Qingsheng LIU ; Jing WEI
Chinese Journal of Pediatrics 2024;62(6):535-541
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).
8.Reference values of carotid intima-media thickness and arterial stiffness in Chinese adults based on ultrasound radio frequency signal: A nationwide, multicenter study
Changyang XING ; Xiujing XIE ; Yu WU ; Lei XU ; Xiangping GUAN ; Fan LI ; Xiaojun ZHAN ; Hengli YANG ; Jinsong LI ; Qi ZHOU ; Yuming MU ; Qing ZHOU ; Yunchuan DING ; Yingli WANG ; Xiangzhu WANG ; Yu ZHENG ; Xiaofeng SUN ; Hua LI ; Chaoxue ZHANG ; Cheng ZHAO ; Shaodong QIU ; Guozhen YAN ; Hong YANG ; Yinjuan MAO ; Weiwei ZHAN ; Chunyan MA ; Ying GU ; Wu CHEN ; Mingxing XIE ; Tianan JIANG ; Lijun YUAN
Chinese Medical Journal 2024;137(15):1802-1810
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.
9.Inhibitory effect of ophiopogonin B on tumor growth in gastric tumor-bearing rats by regulating Rho A/ROCK1 signaling pathway
Mingxing XU ; Ziyin LI ; Hongmei JIANG
Journal of Army Medical University 2024;46(19):2180-2187
Objective To investigate the effect of ophiopogonin B(OP-B)on tumor growth in gastric tumor-bearing rats and its underlying mechanism.Methods Sixty-six SPF SD rats(half male and female,6 weeks old,180~200 g)were randomly divided into model group,low-,medium-and high-dose OP-B groups,and activator group,with 10 rats in each group.The gastric tumor-bearing model was established by orthotopic transplantation of Walker-256 tumor tissue.The rats in the 3 doses groups were given 17.5,35 and 70 mg/kg OP-B,respectively,by gavage and intraperitoneal injection of the same amount of normal saline,and the rats in the activator group were intragastrically administered with 70 mg/kg OP-B and intraperitoneally injected with 1 mg/kg Rho A/ROCK1 signaling pathway activator,lysophosphatidic acid(LPA).The weight and volume of transplanted tumor were recorded to calculate the tumor inhibitory rate.The morphology of tumor tissue was observed with HE staining.The apoptosis of tumor cells were detected by TdT-mediated dUTP-biotin nick end labeling(TUNEL)staining.The expression of proliferating cell nuclear antigen 67(Ki-67)and cleavage cysteine aspartic acid proteolytic enzyme-3(Cleaved Caspase-3)in tumor tissues were detected by immunohistochemical staining.RT-qPCR and Western blotting were utilized to measure the expression of Ras homologous gene family member A(Rho A)and Rho-associated coiled-coil forming protein kinasel(ROCK1)at mRNA and protein levels.Results Compared with the model group,the tumor weight and volume,mRNA and protein levels of Ki-67,Rho A and ROCK1 were significantly decreased,and the tumor inhibitory rate,tumor cell apoptotic rate and Cleaved Caspase-3 protein level were obviously increased in the low-,medium-and high-dose OP-B groups(P<0.05).The activator group had heavier tumor weight and larger volume,increased mRNA and protein expression levels of Ki-67,Rho A and ROCK1,and lower tumor inhibitory rate and apoptotic rate and reduced Cleaved Caspase-3 expression when compared with the high-dose OP-B group(P<0.05).Conclusion OP-B may suppress the tumor growth of gastric tumor-bearing rats by inhibiting the activation of Rho A/ROCK1 signaling pathway.
10.Residual coronary artery tree description and lesion EvaluaTion (CatLet) score, clinical variables, and their associations with outcome predictions in patients with acute myocardial infarction.
Mingxing XU ; Shu WANG ; Ying ZHANG ; Jie ZHANG ; Jin MA ; Junfei SHEN ; Yida TANG ; Tingbo JIANG ; Yongming HE
Chinese Medical Journal 2023;136(20):2459-2467
BACKGROUND:
We have recently developed a new Coronary Artery Tree description and Lesion EvaluaTion (CatLet) angiographic scoring system. Our preliminary studies have demonstrated its superiority over the the Synergy between percutaneous coronary intervention (PCI) with Taxus and Cardiac Surgery (SYNTAX) score with respect to outcome predictions for acute myocardial infarction (AMI) patients. The current study hypothesized that the residual CatLet (rCatLet) score predicts clinical outcomes for AMI patients and that a combination with the three clinical variables (CVs)-age, creatinine, and ejection fraction, will enhance its predicting values.
METHODS:
The rCatLet score was calculated retrospectively in 308 consecutively enrolled patients with AMI. Primary endpoint, major adverse cardiac or cerebrovascular events (MACCE) including all-cause mortality, non-fatal AMI, transient ischemic attack/stroke, and ischemia-driven repeat revascularization, was stratified according to rCatLet score tertiles: rCatLet_low ≤3, rCatLet_mid 4-11, and rCatLet_top ≥12, respectively. Cross-validation confirmed a reasonably good agreement between the observed and predicted risks.
RESULTS:
Of 308 patients analyzed, the rates of MACCE, all-cause death, and cardiac death were 20.8%, 18.2%, and 15.3%, respectively. Kaplan-Meier curves for all endpoints showed increasing outcome events with the increasing tertiles of the rCatLet score, with P values <0.001 on trend test. For MACCE, all-cause death, and cardiac death, the area under the curves (AUCs) of the rCatLet score were 0.70 (95% confidence intervals [CI]: 0.63-0.78), 0.69 (95% CI: 0.61-0.77), and 0.71 (95% CI: 0.63-0.79), respectively; the AUCs of the CVs-adjusted rCatLet score models were 0.83 (95% CI: 0.78-0.89), 0.87 (95% CI: 0.82-0.92), and 0.89 (95% CI: 0.84-0.94), respectively. The performance of CVs-adjusted rCatLet score was significantly better than the stand-alone rCatLet score in terms of outcome predictions.
CONCLUSION:
The rCatLet score has a predicting value for clinical outcomes for AMI patients and the incorporation of the three CVs into the rCatLet score will enhance its predicting ability.
TRIAL REGISTRATION
http://www.chictr.org.cn , ChiCTR-POC-17013536.
Humans
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Coronary Artery Disease/complications*
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Death
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Myocardial Infarction/etiology*
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Percutaneous Coronary Intervention
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Retrospective Studies
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Risk Assessment
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Risk Factors
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Treatment Outcome

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