1.Development and evaluation of a competitive ELISA based on a porcine neutralizing Fab antibody against Senecavirus A.
Yubin LIANG ; Xueqing MA ; Yixuan HE ; Caihe WANG ; Kun LI ; Pinghua LI ; Yuanfang FU ; Zengjun LU ; Xiaohua DU ; Xia LIU ; Pu SUN
Chinese Journal of Biotechnology 2025;41(7):2748-2759
Senecavirus A (SVA) is a major viral pathogen causing disease in pigs, and effective monitoring of SVA infection is critical for disease control. In this study, we aimed to develop a reliable ELISA method for rapidly detecting neutralizing antibodies against SVA. We used HEK293F cells to express an SVA-specific porcine Fab antibody and verified the biological activity of the Fab antibody by indirect ELISA, immunofluorescence assay, virus neutralization test, and Western blotting. The Fab antibody was biotinylated and used as a competitive antibody to establish a competitive ELISA (C-ELISA) for detecting neutralizing antibodies against SVA. We then evaluated the C-ELISA in terms of sensitivity, specificity, repeatability, and result agreement rate with the VNT. The results showed that we successfully prepared an SVA-specific porcine Fab antibody, which showed high affinity for SVA. We named this antibody 1M33Fab and designated it as Bio-1M33Fab after biotin labeling. The assay conditions were optimized as follows: the coating concentration of SVA particles being 1 μg/mL, the working concentration of Bio-1M33Fab being 0.5 μg/mL, the optimal serum dilution of 1:10, and the optimal dilution of enzyme-labeled avidin being 1:30 000. At a percent inhibition (PI) of 47%, the assay demonstrated the highest sensitivity (96.88%) and specificity (100%), with no cross-reactivity observed with the positive sera of major porcine viral diseases. The intra-assay coefficient of variation ranged from 1.12% to 7.34%, while the inter-assay coefficient of variation ranged from 1.10% to 8.97%, indicating good repeatability. In the detection of 224 clinical pig serum samples, C-ELISA and VNT showed a result agreement rate of 93.75%. In conclusion, we successfully develop a C-ELISA method for detecting neutralizing antibodies against SVA by using a porcine-derived Fab antibody, which lays a foundation for the development of detection kits.
Animals
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Swine
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Antibodies, Neutralizing/immunology*
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Enzyme-Linked Immunosorbent Assay/methods*
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Immunoglobulin Fab Fragments/immunology*
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Antibodies, Viral/immunology*
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Picornaviridae/immunology*
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Humans
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HEK293 Cells
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Swine Diseases/diagnosis*
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Picornaviridae Infections/diagnosis*
2.Analyses of varicella epidemic trends in Taizhou, Zhejiang Province, 2014‒2023
Mingming GU ; Qiang CHENG ; Hongbiao LIANG ; Xueqing ZHANG
Shanghai Journal of Preventive Medicine 2025;37(9):742-745
ObjectiveTo analyze the epidemiological characteristics and trends of varicella in Taizhou, Zhejiang Province from 2014 to 2023, and to provide evidence for surveillance, prevention, and control of varicella in Taizhou
3.Effects of extreme temperature exposure on other infectious diarrhea morbidity risk in Taizhou, Zhejiang
Mingming GU ; Qiang CHENG ; Hongbiao LIANG ; Xueqing ZHANG ; Feixiang XU
Journal of Environmental and Occupational Medicine 2025;42(12):1422-1428
Background Other infectious diarrhea (OID) refers to infectious diarrhea other than cholera, dysentery, and typhoid/paratyphoid fever. In the global climate change context, frequent extreme temperature events pose a significant threat to population health, yet the association between extreme temperatures and OID remains unclear. Objective To understand the impact of extreme temperature events on the incidence of OID in Taizhou region, and provide a scientific basis for formulating targeted public health intervention strategies. Methods Case data of OID from 2016 to 2024 and meteorological and pollutant data of Taizhou, Zhejiang during the same period were collected. The case data were derived from National Notifiable Infectious Diseases Reporting System, while the meteorological data and pollutant data were obtained from National Meteorological Science Data Center and Taizhou Ecology and Environment Bureau, respectively. An extreme heat event is defined as a period of two or more consecutive days in the warm season during which the daily mean temperature exceeds the 90th percentile value for that season. Similarly, an extreme cold event is defined as a period of two or more consecutive days in the cold season during which the daily mean temperature falls below the 10th percentile value. A distributed lag nonlinear model based on Poisson regression was used to assess the impact of extreme temperatures on OID incidence. Furthermore, subgroup analyses were conducted by gender and age. Results From 2016 to 2024, a total of 108 670 OID cases were reported in Taizhou, with higher incidence in males (61 253 cases) than females (47 417 cases). In terms of single-day lag effect, exposure to extreme cold presented an inverted U-shaped risk pattern: a significant risk effect began to emerge on the 4th day after exposure, continued to rise, and reached a peak on the 7th day (RR=1.044, 95%CI: 1.029, 1.058), whereas extreme heat had no significant impact. In terms of cumulative lag effect, exposure to extreme cold presented a J-shaped cumulative effect curve: short-term exposure (0-2 days) showed no risk effect, but as time extended (0-9 days), a risk effect emerged and reached the maximum risk value during the 0-12 d cumulative period (RR=1.243, 95%CI: 1.057, 1.461). In contrast, extreme heat had no cumulative lag effect. The subgroup analysis revealed that under extreme cold exposure, the single-day lag effects for both males and females exhibited an inverted U-shaped curve, with the risk peaking on day 7 for males (RR=1.045, 95%CI: 1.026, 1.065) and day 8 for females (RR=1.041, 95%CI: 1.019, 1.064). The risk pattern in the 1-5 years age group was similar to that of the overall population but with a higher effect size (RR=1.063, 95%CI: 1.041, 1.085). In terms of the cumulative lag effects, the cumulative risk curves for both males and females under extreme cold exposure showed a J-shaped pattern with varied risk profiles. Significant variations in risk patterns were observed across different age groups. No significant cumulative lag effects were observed for extreme heat exposure in any subgroup. Conclusion Extreme cold can increase the incidence of OID in Taizhou, and the risk effects are heterogeneous among different groups. No risk effect is observed for extreme heat.
4.Fire safety management in large public hospitals from the perspective of collaborative governance
Huan MA ; Yushui TANG ; Chengliang WANG ; Xueqing LIANG
Modern Hospital 2024;24(8):1269-1273
Fire safety is crucial for the regular operation of large public hospitals.This paper elaborates on the funda-mental guidelines for fire safety management in large hospitals.Under the framework of collaborative governance theory,it also analyzes and points out that the standardization of peoples behavior in hospitals and the integrity of fire safety facilities are the pri-mary factors influencing the efficacy of fire safety management.Subsequently,based on management practices,it suggests focu-sing on"five synergies"in enforcing primary responsibility for fire safety,namely,the implementation of fire safety main respon-sibilities in synergy,the regular inspection and rectification of fire hazards in synergy,the training and evaluation of basic fire safety skills in synergy,the lifecycle management of fire safety facilities in synergy,and the building of a fire safety management team in synergy.This approach aims to maximize overall fire safety effectiveness and comprehensively enhance the level of fire safety management in hospitals.Finally,the paper discusses and calls for further enhancements in the management of fire safety in large public hospitals.
5.Prognosis and risk factors of IgA vasculitis nephritis in children
Xueqing MA ; Yonghua HE ; Jinyun PU ; Wenpei LIANG ; Panpan SHAO ; Jianhua ZHOU ; Yu ZHANG ; Jinhui TANG ; Tonglin LIU ; Huiqing YUAN ; Liru QIU
Chinese Journal of Pediatrics 2024;62(12):1184-1190
Objective:To investigate the prognosis and risk factors of IgA vasculitis nephritis (IgAVN) in children.Methods:A retrospective cohort study was conducted. Clinical data were collected from 264 children who were pathologically diagnosed with IgAVN at Department of Pediatric Nephrology, Tongji Hospital, affiliated with Tongji Medical College, Huazhong University of Science and Technology, between January 2011 and December 2017. All patients had a follow-up period of more than 3 years. Clinical characteristics, renal pathology, 3-year and 5-year prognosis were analyzed. The patients were grouped based on gender, age of onset (≤6 years, >6-9 years, and >9 years), pathological classification (≤Ⅲ and>Ⅲ),whether the prognosis was complete remission at 3 and 5 years. Independent sample t-tests, ANOVA or chi-squared test were used for intergroup comparisons. Spearman correlation analysis was applied for ordinal data, and multivariate Logistic regression was used to analyze factors affecting the prognosis. Receiver operating characteristic (ROC) curve was utilized to evaluate the predictive value of these factors. Results:Of the 264 children with IgAVN, 153 were male and 111 were female, the age of onset was 8.3 (6.7, 10.3) years, 118 patients (45%) with onset age >6-9 years accounted for the highest proportion. All patients presented with skin purpura and renal involvement, primarily manifesting as hematuria and/or proteinuria. Microscopic hematuria was observed in 253 patients (95.8%), while 246 patients (93.2%) showed proteinuria. In 256 patients (97.0%), hematuria or proteinuria urinalysis was detected within 6 months of skin purpura onset, and 243 patients (92.0%) underwent renal biopsy within 6 months of renal involvement. The most common clinical subtype in 264 IgAVN children was hematuria and proteinuria (204 cases, 77.3%), with grade Ⅲ being the predominant pathological classification (181 cases, 68.6%). Among children ≤6 years old, the 3-year complete remission rate was higher in males than in females (83.9% (26/31) vs. 7/16, χ2=8.12, P=0.012). Factors independently associated with poor 5-year prognosis included time from hematuria or proteinuria urinalysis to renal biopsy >6 months, elevated serum cholesterol levels, and incomplete remission 3 years post-biopsy ( OR=5.41, 1.39, 6.02, 95% CI 1.40-20.86, 1.04-1.84, 2.61-13.88, all P<0.05). The serum cholesterol has a predictive value for 5-year prognosis ( P=0.020, AUC=0.62, 95% CI 0.52-0.71, Youden index=0.27, cutoff=4.37). Conclusions:For children with IgAVN aged≤6 years, the 3-year prognosis is better in males than in females. Time from hematuria or proteinuria urinalysis to renal biopsy >6 months, elevated serum cholesterol levels, and incomplete remission at 3 years post-biopsy may be independent risk factors for poor 5-year prognosis in children with IgAVN.
6.Analysis of clinical phenotype and genotype of PAX2 mutation in Chinese children
Xueqing MA ; Yonghua HE ; Jing YANG ; Rongrong XU ; Siying YANG ; Wenpei LIANG ; Jianhua ZHOU ; Huiqing YUAN ; Liru QIU
Chinese Journal of Nephrology 2024;40(1):24-35
Objective:To analyze and summarize the clinical, genotypic and pathological characteristics of children with PAX2 gene mutation in China, and to provide information for the monitoring, treatment and prognosis of the disease. Methods:It was a case series analysis study. The clinical data of children with PAX2 gene mutation in Pediatric Nephrology Department, Tongji Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology from January 2014 to December 2022 were collected, and peripheral blood gene DNA was extracted and sequenced for whole exome sequencing. The clinical, pathological and genotypic characteristics of PAX2 gene variation of children in China were summarized by searching PubMed, Medline, China National Knowledge Infrastructure and Wanfang database and compared with the cases in this single center. Results:Among the 13 children with PAX2 gene mutation, there were 9 males and 4 females, 12 patients with abnormal urine tests, 7 patients with small kidney volume by imaging examination, and 5 patients with renal cysts. The clinical phenotypes were congenital renal and urinary tract malformations in 8 cases, renal coloboma syndrome in 1 case, and hematuria or proteinuria in 3 cases. Five patients underwent renal biopsies, showing focal segmental glomerulosclerosis and C3 glomerulopathy in 1 case, focal segmental glomerulosclerosis in 1 case, thin basement membrane lesion in 1 case, and IgA nephropathy in 2 cases. The genetic testing in 13 children showed 9 de novo mutations and 4 new mutations of c.321G>A, c.213-8C>G, c.63C>A and c.449C>T. There were 2 cases of 76dupG (p.V26Gfs*28) mutant. A total of 51 Chinese children with PAX2 gene mutation were found in the literature search. There were 32 males and 19 females, 8 cases with small kidney volume and 12 cases with renal cysts. The clinical phenotypes were congenital anomalies of kidney and urinary tract in 28 cases, renal coloboma syndrome in 17 cases, and hematuria or proteinuria in 6 cases. Seven patients underwent renal biopsies, including 2 cases with focal segmental glomerulosclerosis, 1 case with minimal lesion, 1 case with mesangial proliferative glomerulonephritis, 1 case with IgA nephropathy, 1 case with membranous nephropathy and a case with focal proliferative sclerosing purpura nephritis combined with glomerular hypertrophy. Thirty-four cases were de novo mutations, and 12 mutations were from the father or mother. The father or mother of 5 children had no clinical manifestations, with normal renal function. There were 11 cases of 76dupG (p.V26Gfs*28) mutant. Conclusions:The clinical phenotypes and genotypes of PAX2 gene variation in Chinese children are diverse. The most common clinical phenotype of PAX2 gene variation is congenital anomalies of kidney and urinary tract. c.76dupG (p.V26Gfs*28) is the most common of PAX2 gene variant.
7.Prediction of peritoneal free cancer cells in gastric cancer patients by golden-angle radial sampling dynamic contrast-enhanced magnetic resonance imaging
YIN XUEQING ; RUAN XINZHONG ; ZHU YONGMENG ; YIN YONGFANG ; HUANG RUI ; LIANG CHAO
Journal of Zhejiang University. Science. B 2024;25(7):617-627
Objective:Peritoneal free cancer cells can negatively impact disease progression and patient outcomes in gastric cancer.This study aimed to investigate the feasibility of using golden-angle radial sampling dynamic contrast-enhanced magnetic resonance imaging(GRASP DCE-MRI)to predict the presence of peritoneal free cancer cells in gastric cancer patients.Methods:All enrolled patients were consecutively divided into analysis and validation groups.Preoperative magnetic resonance imaging(MRI)scans and perfusion were performed in patients with gastric cancer undergoing surgery,and peritoneal lavage specimens were collected for examination.Based on the peritoneal lavage cytology(PLC)results,patients were divided into negative and positive lavage fluid groups.The data collected included clinical and MR information.A nomogram prediction model was constructed to predict the positive rate of peritoneal lavage fluid,and the validity of the model was verified based on data from the verification group.Results:There was no statistical difference between the proportion of PLC-positive cases predicted by GRASP DCE-MR and the actual PLC test.MR tumor stage,tumor thickness,and perfusion parameter Tofts-Ketty model volume transfer constant(Ktrans)were independent predictors of positive peritoneal lavage fluid.The nomogram model featured a concordance index(C-index)of 0.785 and 0.742 for the modeling and validation groups,respectively.Conclusions:GRASP DCE-MR could effectively predict peritoneal free cancer cells in gastric cancer patients.The nomogram model constructed using these predictors may help clinicians to better predict the risk of peritoneal free cancer cells being present in gastric cancer patients.
8.Feature analysis of the tissue methylation profile in adenocarcinoma patients with pulmonary nodules on CT scan
Qiaomei GUO ; Lihua QIAO ; Lin WANG ; Xueqing WANG ; Fei WU ; Xiaohui LIANG ; Yuteng SUN ; Jiatao LOU
Chinese Journal of Laboratory Medicine 2024;47(11):1277-1285
Objective:To investigate the tissue methylation features of adenocarcinoma patients presenting as pulmonary nodules on CT scans.Methods:A retrospective analysis was conducted on 70 adenocarcinoma patients with pulmonary nodules diagnosed at the Shanghai General Hospital from June 1, 2022 to January 20, 2024. Participants were assigned to two groups using the random number table, with 40 in the discovery group and 30 in the validation group. In the discovery group, tissue samples were analyzed using reduced representation bisulfite sequencing (RRBS) technology to compare the average methylation levels between cancer tissues and paired adjacent non-cancerous tissues. Differentially methylated regions (DMRs) were screened for analysis of their distribution across various genomic functional elements, and hierarchical clustering was plotted. GO and KEGG pathway enrichment analyses were further conducted on the DMRs. Subsequently, candidate DMRs associated with lung adenocarcinoma were validated using TCGA lung adenocarcinoma cohort and targeted bisulfite sequencing technology in the validation group. The comparison of methylation levels between groups was conducted using t-tests or non-parametric tests, while rates and composition ratios were analyzed using chi-square tests or Fisher′s exact test.Results:In discovery cohort, the average methylation level in cancer tissues was lower compared to adjacent normal tissues [(42.369±4.627) vs (44.370±4.046), t=?2.059, P=0.043]. A total of 37 995 DMRs were identified, including 16 889 upregulated regions and 21 106 downregulated regions, predominantly locating in promoter regions (48.917%), introns (36.457%), and exons (10.812%). The DMR clustering heatmap revealed two distinct clusters corresponding to cancer tissues and adjacent non-cancerous tissues. GO analysis showed that DMRs associated genes were mainly located in the cell membrane and nuclear chromatin, and were primarily involved in RNA polymerase Ⅱ-related transcription and regulation. KEGG pathway enrichment analysis indicated that DMRs associated genes were mainly involved in neuroactive ligand-receptor interaction, cancer pathways, calcium signaling pathway, cAMP signaling pathway, and MAPK signaling pathway. Validation in the TCGA cohort confirmed 11 potential characteristic DMRs. In the validation group, TBS confirmed that the methylation levels of DMRs associated with MIR10B, DMRTA2, HOPX, TFAP2B and MARCH11 in cancer tissues were significantly higher than those in adjacent non-cancerous tissues [11.200(4.305, 27.088) vs 2.650(1.298, 4.645), Z=?4.539, P<0.05; 18.610(13.600, 33.025) vs 8.675(5.488, 13.085), Z=?4.554, P<0.05; 17.600(2.183, 76.015) vs 1.085(0.898, 1.835), Z=?5.131, P<0.05; 5.250(3.220, 7.693) vs 3.495(2.165, 4.383), Z=?2.861, P<0.05; 11.515(7.525, 21.033) vs 7.830(5.518, 11.488), Z=?2.440, P<0.05 ], and the differences were statistically significant. Conclusions:Lung adenocarcinoma tissue exhibits different methylation patterns compared with adjacent normal lung tissue. The identified DMRs are involved in the regulation of several key pathways. Results from the TCGA cohort and an independent validation group support the potential diagnostic value of DMRs such as MIR10B, DMRTA2, HOPX, TFAP2B, and MARCH11 in lung adenocarcinoma, though their clinical application requires further validation.
9.Application of a modified pancreatogastric anastomosis in laparoscopic duodenum-preserving pancreatic head resection
Jianzhang QIN ; Haotian YU ; Xueqing LIU ; Xinbo ZHOU ; Wei HE ; Yunfei LIANG ; Qing ZHANG ; Jianhua LIU
Chinese Journal of Hepatobiliary Surgery 2023;29(12):927-931
Objective:To study the feasibility of a modified pancreatogastric anastomosis in laparoscopic duodenum preserving pancreatic head resection (LDPPHR).Methods:The clinical data of 25 patients with benign or low-grade malignant tumors of pancreatic head undergoing LDPPHR at the Second Hospital of Hebei Medical University from January 2019 to May 2023 were retrospectively analyzed, including 7 males and 18 females, aged (44.9±6.2) years old. According to the methods of pancreatic digestive reconstruction, patients were divided into the observation group ( n=10), who underwent the modified pancreatogastric anastomosis, and the control group ( n=15) who underwent conventional pancreaticojejunal anastomosis and jejuno-jejunal anastomosis. The general data, intraoperative pancreatic digestive reconstruction time, maximum levels of amylase in abdominal drainage within three days postoperatively, postoperative complications, and hospital stay were compared between the groups. Results:All procedures were performed successfully. The intraoperative pancreatic digestive reconstruction time was shorter in the observation group [(27.8±2.4) min vs. (45.8±3.6) min, P=0.010]. The intraoperative blood loss were comparable between the groups [(140.5±14.8) ml vs. (145.2±9.7) ml, P=0.843]. The maximum level of amylase in abdominal drainage within three days postoperatively was lower in the observation group [(809.1±185.5) U/L vs. (1 385.4±481.1) U/L, P=0.031]. No grade C pancreatic fistula or postoperative hemorrhage occurred in either group, and the incidence of grade B pancreatic fistula was lower in the observation group [20.0% (2/10) vs. 60.0% (9/15), P=0.048], with a shorter postoperative hospital stay [(7.9±1.3) d vs. (10.3±2.7) d, P=0.017]. No decrease of life quality or reoperation due to pancreatic fistula, hemorrhage or digestive tract malfunction occurred in either group within a median follow-up of 15.6 months. Conclusion:In LDPPHR, the modified pancreatogastric anastomosis could help shorten the pancreatic digestive reconstruction and lower the risk of postoperative pancreatic fistula.
10.Research progress of risk prediction model for peripherally inserted central catheter related thrombosis in patients with malignant tumor
Shengmiao MA ; Lei WANG ; Shengxiao NIE ; Pei WANG ; Xiaoxia WANG ; Xueqing PENG ; Peng LIU ; Yong LAN ; Tao LIANG ; Hong SUN
Chinese Journal of Modern Nursing 2023;29(17):2355-2360
Previous studies have comprehensively evaluated predictive models related to peripherally inserted central catheter-related venous thrombosis (PICC-RVT) , but have not further elaborated on its application and effect in the tumor patient population. This article reviews the PICC-RVT evaluation tools that are clinically mature and applicable to cancer patients, analyzes the presentation forms, statistical methods, and limitations of different models, in order to provide suggestions and references for model selection by clinical healthcare professionals and the construction of future PICC-RVT prediction models for cancer patients.

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