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.TSR2 overexpression inhibits proliferation and invasion of gastric cancer cells by downregulating the PI3K/AKT signaling pathway
Yongsheng XIA ; Lian WANG ; Xiaohua CHEN ; Yulu ZHANG ; Aofei SUN ; Deli CHEN
Journal of Southern Medical University 2024;44(5):913-919
Objective To investigate the expression of TSR2 in gastric cancer and explore its correlation with progression of gastric cancer and the possible mechanism.Methods We retrospectively analyzed TSR2 expression in clinical specimens from 105 gastric cancer patients and the impact of TSR2 expression level on disease progression and 5-year postoperative survival of the patients.GO and KEGG enrichment analyses were used to predict the biological functions and mechanisms of TSR2.In gastric cancer MGC-803 cells with lentivirus-mediated TSR2 overexpression or knockdown,the changes in cell proliferation,invasion,and migration were assessed with CCK-8 and Transwell assays,and the expressions of p-PI3K and p-AKT were detected using Western blotting.Results TSR2 expression was significantly lower in gastric cancer tissues than in the adjacent tissues with significant correlations with CEA level,CA19-9 level,and T and N staging(P<0.05).A low TSR2 expression,CEA≥5 μg/L,CA19-9≥37 kU/L,T3-T4 stages,and N2-N3 staged were identified as independent risk factors affecting 5-year survival rate of the patients following radical surgery(P<0.05),and a high TSR2 expression was associated with a higher 5-year survival rate of the patients(P<0.001).Bioinformatics analysis suggested the functional involvement of TSR2 with the PI3K/AKT signaling pathway.MGC-803 cells overexpressing TSR2 showed significantly lowered proliferation,migration,and invasion capacities(P<0.05),while TSR2 knockdown produced the opposite effects(P<0.05).Western blotting showed that TSR2 overexpression reduced the phosphorylation of PI3K and AKT,and TSR2 knockdown caused the opposite changes in MGC-803 cells(P<0.05).Conclusion TSR2 is lowly expressed in gastric cancer tissues to adversely affect the patients'prognosis,and its overexpression inhibits gastric cancer cell proliferation,invasion,and migration possibly by downregulating the PI3K/AKT pathway.
3.Expert consensus on the test development and preliminary implementation of whole genome sequencing for fetal structural abnormalities
Yanfei WANG ; Xiaofan ZHU ; Luming SUN ; Xiaohua TANG ; Ning LIU ; Xiangdong KONG
Chinese Journal of Medical Genetics 2024;41(6):677-684
Fetal structural anomalies and birth defects are primarily caused by genetic variants such as chromosomal number abnormalities, copy number variations (CNV), single nucleotide variants (SNV), and small insertions and deletions (indel). Whole-genome sequencing (WGS) based on next-generation sequencing (NGS) as an emerging technology for genetic disease diagnosis can detect the aforementioned types of variants. In recent years, high-depth WGS (> 30×) for prenatal diagnosis has also become available, and proved to be practical for unraveling the genetic etiology of fetal developmental abnormalities. To fascilitate clinical practice, test development and preliminary implementation of WGS for diagnosing fetal structural anomalies, we have formulated a consensus over the application of WGS in prenatal diagnosis by compiling previously published consensuses, guidelines, and research findings to provide a guidance on data analysis, reporting recommendations, and consultation of prenatal WGS results.
4.Study on the application of nursing intervention based on mindfulness-attention-acceptance theory in children with bronchopneumonia
Xiaohua SUN ; Xueyan YUE ; Xumei CHEN ; Rui MA ; Liang RU
Chinese Journal of Modern Nursing 2024;30(26):3621-3625
Objective:To explore the application effect of nursing intervention based on the mindfulness-attention-acceptance theory in children with bronchopneumonia.Methods:Totally 127 children with bronchopneumonia admitted to the First Affiliated Hospital of Xinjiang Medical University from January to July 2023 were selected by convenience sampling. They were randomly divided into a control group and an observation group according to a random number table. The control group (63 cases) received routine nursing intervention, while the observation group (64 cases) received nursing intervention based on the mindfulness-attention-acceptance theory in addition to the routine nursing intervention. The improvement in psychological resilience, coping styles, and emotional behaviors of the two groups were compared before and after the intervention.Results:After the intervention, the scores of all dimensions of the Psychological Resilience Scale in both groups were higher than that before the intervention, with the observation group scoring higher than the control group ( P<0.05). The observation group scored higher on all dimensions of positive coping and lower on all dimensions of negative coping compared to the control group ( P<0.05). The prosocial behavior dimension score of the observation group was higher than that of the control group, while the scores of emotional symptoms, conduct problems, and hyperactivity-inattention dimensions were lower than those of the control group ( P<0.05) . Conclusions:The application of nursing intervention based on the mindfulness-attention-acceptance theory in children with bronchopneumonia can effectively improve their psychological resilience, coping styles, and emotional behaviors.
5.Psychometric properties of the Chinese version of the Pediatric Epilepsy-Specific Health-Related Quality of Life in children aged 5 to 7 years old and their parents
Yiming ZHANG ; Anwei XIE ; Xiaohua WANG ; Li TIAN ; Lu LIN ; Lihua SUN ; Feng MIAO ; Xuan ZHAO
Chinese Journal of Modern Nursing 2024;30(30):4096-4104
Objective:To translate the Pediatric Epilepsy-Specific Health-Related Quality of Life (PedsQL TM Epilepsy Module) into Chinese and test its reliability and validity in children aged 5 to 7 years old and their parents. Methods:Brislin translation model was used for forward translation, back translation, harmonization, and proofreading of the PedsQL TM Epilepsy Module. Chinese version of PedsQL TM Epilepsy Module beta version was developed through cognitive interviews, expert consultations, and pre-surveys for cross-cultural adaptation of the scale. From April to November 2023, convenience sampling was used to select 420 children with epilepsy and their parents admitted to the Children's Hospital of Soochow University as participants for a questionnaire survey to test its reliability and validity. Results:A total of 840 questionnaires were distributed (420 for children and 420 for parents), and 394 valid questionnaires were collected from children and 400 valid questionnaires were collected from parents, and the effective response rates of the questionnaires were 93.81% (394/420) and 95.24% (400/420), respectively. Chinese version of PedsQL TM Epilepsy Module (child report version) included five dimensions and 28 items. Chinese version of PedsQL TM Epilepsy Module (parent agent report version) consisted of five dimensions and 29 items. The content validity indices at the item level were 0.800 to 1.00, while the content validity indices at the scale level were 0.978 and 0.979, respectively. Exploratory factor analysis of two versions of the scale showed that five common factors were extracted, with cumulative variance contribution rates of 64.557% and 75.205%, respectively. Confirmatory factor analysis showed that the models of the two scales fitted well. The total Cronbach's α coefficients of the two scales were 0.906 and 0.914, respectively, with Cronbach's α coefficients for each dimension ranging from 0.869 to 0.991. The total test-retest reliability coefficients of the scale were 0.998 and 0.995, respectively, and the test-retest reliability coefficients of each dimension were 0.803 to 0.995. Conclusions:Chinese version of the PedsQL TM Epilepsy Module has good reliability and validity, and is suitable for evaluating the health-related quality of life of children with epilepsy aged 5 to 7 years old in China.
6.A review of research advances in virtual reality-based interventions for children with autism
Yu FANG ; Wenhao LIN ; Xiaohua SUN ; Yuan ZHAO ; Liying DONG ; Haidong SONG
Chinese Mental Health Journal 2024;38(5):394-399
This paper analyzes the literature on virtual reality(VR)technology interventions for autistic children published from 2014 to 2022.It shows that different VR systems in these studies have improved different symptoms of autism,and points out the feasibility of VR technology interventions for autistic children.
7.Characteristics of anger emotions in patients with schizophrenia with violent behavior in community
Mingjin LUO ; Kejie WANG ; Pingyu TAO ; Xiaohua SUN ; Haidong SONG
Chinese Mental Health Journal 2024;38(10):840-845
Objective:To explore the anger emotional state,traits and expression characteristics in patients with schizophrenia with violent behavior in community.Methods:Twenty-five patients with schizopnrenia with a history of violent behavior among the community rehabilitation institutions were selected as the violent behavior group,and 74 patients with schizopnrenia without a history of violent behavior were selected as the nonviolent behavior group.The Brief Psychiatric Rating Scale(BPRS)was used to assess psychiatric symptoms,and the State-Trait An-ger Expression Inventory(STAXI)was used to assess anger characteristics.Results:Compared with the nonviolent behavior group,the violent behavior group had significantly higher scores in the trait of anger(patient dimension score,rational dimension score)and expression of anger(total score,restraint dimension score)[(28.0±23.8)vs.(19.6±13.3),(55.0±30.6)vs.(40.0±21.5),(34.2±11.6)vs.(27.3±10.7),(56.0±26.1)vs.(33.7±21.1),Ps<0.05].Higher scores of rationality dimension of anger trait(OR=3.69,95%CI:1.43-7.98)and re-straint dimension of anger expression(OR=3.25,95%CI:1.39-7.47)were risk factors for violent behaviors.Conclusion:Compared with patients with schizopnrenia without violent behavior,patients with violent behavior history may show more lack of rationality,and poorer anger restraint.
8.TSR2 overexpression inhibits proliferation and invasion of gastric cancer cells by downregulating the PI3K/AKT signaling pathway
Yongsheng XIA ; Lian WANG ; Xiaohua CHEN ; Yulu ZHANG ; Aofei SUN ; Deli CHEN
Journal of Southern Medical University 2024;44(5):913-919
Objective To investigate the expression of TSR2 in gastric cancer and explore its correlation with progression of gastric cancer and the possible mechanism.Methods We retrospectively analyzed TSR2 expression in clinical specimens from 105 gastric cancer patients and the impact of TSR2 expression level on disease progression and 5-year postoperative survival of the patients.GO and KEGG enrichment analyses were used to predict the biological functions and mechanisms of TSR2.In gastric cancer MGC-803 cells with lentivirus-mediated TSR2 overexpression or knockdown,the changes in cell proliferation,invasion,and migration were assessed with CCK-8 and Transwell assays,and the expressions of p-PI3K and p-AKT were detected using Western blotting.Results TSR2 expression was significantly lower in gastric cancer tissues than in the adjacent tissues with significant correlations with CEA level,CA19-9 level,and T and N staging(P<0.05).A low TSR2 expression,CEA≥5 μg/L,CA19-9≥37 kU/L,T3-T4 stages,and N2-N3 staged were identified as independent risk factors affecting 5-year survival rate of the patients following radical surgery(P<0.05),and a high TSR2 expression was associated with a higher 5-year survival rate of the patients(P<0.001).Bioinformatics analysis suggested the functional involvement of TSR2 with the PI3K/AKT signaling pathway.MGC-803 cells overexpressing TSR2 showed significantly lowered proliferation,migration,and invasion capacities(P<0.05),while TSR2 knockdown produced the opposite effects(P<0.05).Western blotting showed that TSR2 overexpression reduced the phosphorylation of PI3K and AKT,and TSR2 knockdown caused the opposite changes in MGC-803 cells(P<0.05).Conclusion TSR2 is lowly expressed in gastric cancer tissues to adversely affect the patients'prognosis,and its overexpression inhibits gastric cancer cell proliferation,invasion,and migration possibly by downregulating the PI3K/AKT pathway.
9.Influencing factors of genotypic drug resistance in people living with human immunodeficiency virus/acquired immunodeficiency syndrome who failed anti-retroviral therapy in Henan Province from 2018 to 2022
Yan SUN ; Zhaoyun CHEN ; Yuqi HUO ; Mengyao FENG ; Jinjin LIU ; Xuan YANG ; Qingxia ZHAO ; Xiaohua ZHANG ; Shuxian ZHAO ; Xue ZHANG ; Yan WANG
Chinese Journal of Infectious Diseases 2024;42(4):219-224
Objective:To analyze the influencing factors of genotypic drug resistance mutations in people living with human immunodeficiency virus and acquired immunodeficiency syndrome(PLWHA) who failed anti-retroviral therapy (ART) in Henan Province, in order to provide a basis for adjusting ART regimens and reducing drug resistance.Methods:PLWHA with virological failure (human immunodeficiency virus (HIV) RNA≥500 copies/mL) after receiving ART for more than 24 weeks were included in Henan Province from January 2018 to December 2022. Baseline CD4 + T lymphocyte counts, ART regimens and other clinical data were collected. HIV-1 gene subtypes and their drug resistance sequence mutations were detected in the Sixth People′s Hospital of Zhengzhou, and the sequences were submitted to the HIV Drug Resistance Interpretation System of Stanford University for comparison of test results. Genotypic drug resistance to nucleotide reverse transcriptase inhibitors (NRTI), non-nucleoside reverse transcriptase inhibitors (NNRTI), protease inhibitors (PI) and integrase inhibitors (INSTI) was determined. Multivariate logistic regression was used to analyze the influencing factors of drug resistance in patients with ART failure. Results:Among 982 PLWHA, the sequences of 899 cases were successfully amplified, and drug resistance was detected in 737 cases, with the drug resistance rate of 81.98%(737/899). The rates of resistance to NRTIs, NNRTIs, PIs and INSTIs were 71.97%(647/899), 79.31%(713/899), 5.23%(47/899) and 2.72%(20/734), respectively.The largest number of those who developed concomitant resistance to two classes of drugs was 588 cases (79.78%), mainly NRTI and NNRTI concomitant resistance in 583 cases (79.10%). There were 99 cases (13.43%) who developed resistance to only one class of drugs, and those who developed concurrent resistance to three classes of drugs were 48 cases (6.51%), and two cases (0.27%) were found to be resistant to all four classes of drugs mentioned above. A total of 10 HIV genotypes were detected, among which subtype B accounted for the most (59.73%(537/899)), followed by circulating recombinant form (CRF)01_AE subtype (21.91%(197/899)) and CRF07_BC subtype (9.45%(85/899)). The risk factors affecting the development of drug resistance were baseline CD4 + T lymphocyte counts, ART regimens and HIV-1 genotypes. The risk of drug resistance in patients with baseline CD4 + T lymphocyte counts <100/μL was 4.55 times (95% confidence interval ( CI) 2.69 to 7.70) higher than patients with CD4 + T lymphocyte counts≥250/μL, the risk of drug resistance in patients using 2NRTIs+ NNRTI regimen was 4.51 times (95% CI 1.75 to 11.63) higer than those using 2NRTIs+ INSTI regimen, and patients infected with B and CRF01_AE subtype was 2.18 times (95% CI 1.10 to 4.29) and 2.70 times (95% CI 1.26 to 5.78) higer than those with CRF07_BC subtype, respectively. Conclusions:The incidence of genotypic drug resistance in PLWHA with ART failure in Henan Province is high. Low baseline CD4 + T lymphocyte counts, 2NRTIs+ NNRTI regimens, and genotype B and CRF01_AE are risk factors for drug resistance in PLWHA.
10.A cross-sectional study of renal injury in human immunodeficiency virus infection/acquired immunodeficiency syndrome patients after antiviral therapy in Henan Province
Xuan YANG ; Zhongfeng CUI ; Chaoyang LIU ; Lin ZHANG ; Quanxi LI ; Yujiao NIU ; Xue ZHANG ; Xiaohua ZHANG ; Zhaoyun CHEN ; Qiong LI ; Jinjin LIU ; Yan SUN
Chinese Journal of Infectious Diseases 2024;42(7):395-402
Objective:To understand the clinical characteristics of human immunodeficiency virus (HIV) infection/acquired immunodeficiency syndrome (AIDS) patients with renal injury after antiviral therapy in Henan Province, and to explore the risk factors of renal injury.Methods:A cross-sectional study was conducted to investigate HIV infection/AIDS patients receiving antiviral therapy in Zhengzhou Sixth People′s Hospital, Anyang Fifth People′s Hospital, Hebi Third People′s Hospital, Luo Yang Zhoushan Hospital and Lankao Central Hospital in Henan Province from April 1 to September 30, 2023. The clinical information including basic data, antiviral therapy regimens and comorbidities, and laboratory test results (blood urea nitrogen, serum creatinine, blood uric acid, urine routine, urine microalbumin, urine α 1-microglobulin (α 1-MG), urine β 2-microglobulin (β 2-MG), urine retinol binding protein (RBP), urine creatinine, HIV viral load, CD4 + T lymphocyte count) were collected. Multivariate binary logistic regression was used to analyze independent risk factors for renal injury. Results:A total of 2 526 HIV infection/AIDS patients were included, with the age of (45.52±14.28) years and 2 156 (85.4%) males. The main route of transmission was sexual transmission (91.6%, 2 314/2 526). The duration of antiviral therapy was 5.00(2.92, 8.00) years. Tenofovir (TDF)+ lamivudine (3TC)+ non-nucleoside reverse transcriptase inhibitors (NNRTI) accounted for 55.3%(1 396/2 526) of the current antiviral therapy regimen. The percentage of HIV viral load <50 copies/mL was 93.0%(2 350/2 526). The CD4 + T lymphocyte count was 476(337, 645)/μL. There were 156 patients (6.2%) complicated with hepatitis B and/or hepatitis C, 205 patients (8.1%) with diabetes, 379 patients (15.0%) with hyperlipidemia, and 189 patients (7.5%) with hyperuricemia. A total of 1 040 patients (41.2%) with renal injury were found through renal function test, including 355 cases (14.1%) with estimated glomerular filtration rate (eGFR) <60 mL/(min·1.73 m 2) or urine protein positive or urine albumin creatine ratio (UACR) ≥30 mg/g, 682 patients (27.0%) with pure tubular injury presented with only positive for urinary α 1-MG, urinary β 2-MG, or urinary RBP. eGFR< 60 mL/(min·1.73 m 2) was found in 71 cases (2.8%), eGFR from 60 to 89 mL/(min·1.73 m 2) was found in 509 cases (20.2%), and eGFR≥90 mL/(min·1.73 m 2) was found in 1 946 cases (77.0%). A total of 138 patients (5.5%) were identified as having combined chronic kidney disease (CKD). Among them, 110 patients (79.7%) were in CKD stages 1 to 2, and 117 patients (84.8%) were in urinary albumin A2 grade. Multivariate analysis of 355 patients with renal injury who had eGFR<60 mL/(min·1.73 m 2) or positive urine protein in urine routine or UACR ≥30 mg/g showed that ages of 50 to 69 years old (odds ratio( OR)=2.189, 95% confidence interval ( CI) 1.333 to 3.596, P=0.002)), ≥70 years old ( OR=5.190, 95% CI 2.912 to 9.248, P<0.001), female ( OR=1.685, 95% CI 1.241 to 2.286, P=0.001), combined opportunistic infection ( OR=2.521, 95% CI 1.567 to 4.056, P<0.001), combined hepatitis B ( OR=1.962, 95% CI 1.110 to 3.467, P=0.020), combined hepatitis C ( OR=1.883, 95% CI 1.043 to 3.400, P=0.036), combined diabetes ( OR=2.703, 95% CI 1.911 to 3.821, P<0.001), using TDF for two to four years ( OR=1.674, 95% CI 1.103 to 2.459, P=0.015), using TDF for greater than or equal to five years ( OR=1.880, 95% CI 1.287 to 2.746, P=0.001), using TDF combined with lopinavir/ritonavir (LPV/r) ( OR=3.610, 95% CI 2.273 to 5.734, P<0.001) and using TDF combined with non-LPV/r ( OR=1.495, 95% CI 1.036 to 2.157, P=0.031) were the risk factors of renal injury. Conclusions:There is a high proportion of renal injury among HIV infection/AIDS patients after antiviral therapy in Henan Province, including CKD and simple renal tubular injury. Older age, female, comorbidities, and long-term use of TDF are risk factors for renal injury.

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