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
;
Swine
;
Antibodies, Neutralizing/immunology*
;
Enzyme-Linked Immunosorbent Assay/methods*
;
Immunoglobulin Fab Fragments/immunology*
;
Antibodies, Viral/immunology*
;
Picornaviridae/immunology*
;
Humans
;
HEK293 Cells
;
Swine Diseases/diagnosis*
;
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.Application of breakthrough series quality improvement model in preventing blood flow infections related to non-cuffed catheters
Na CHEN ; Yushen REN ; Li TIAN ; Xiaoping WANG ; Yujun WANG ; Yanling SUN ; Hongwen MA ; Xiaohua YUAN
Chinese Journal of Practical Nursing 2024;40(21):1601-1607
Objective:To explore the application effect of breakthrough series (BTS) quality improvement model in the prevention of catheter-related bloodstream infection in hospitalized patients with indwelling non-cuffed catheter (NCC).Methods:Using a non synchronous pre and post control study method, convenience sampling was used to select NCC patients from four hospitals in Tianjin from January to September 2022 who received conventional nursing plans as the control group, and NCC patients from February to October 2023 who received nursing plans based on the BTS quality improvement model as the observation group. Compared the incidence of NCC related bloodstream infections between two groups of patients, the implementation of key preventive measures for NCC related bloodstream infections by nursing staff, and patient satisfaction.Results:Among the 984 patients included in the control group, there were 687 males and 297 females, aged (62.43 ± 13.77) years old; among the 959 patients included in the observation group, there were 651 males and 308 females, aged (61.96 ± 13.89) years old. After applying the improved model, the incidence of NCC related bloodstream infections in the observation group was 0.12‰ (1/8 676), lower than the control group′s 0.71‰ (7/9 827), and the difference was statistically significant ( χ 2=4.37, P<0.05) ;the implementation rate of key measures for preventing NCC related bloodstream infections in the observation group was 90.00% (54/60) for catheter outlet care and 91.67% (55/60) for maximizing sterile barrier, both higher than 70.37% (38/54) and 75.93% (41/54) in the control group, with statistical significance ( χ2=7.03, 5.30, both P<0.05); the total satisfaction rate of patients in the observation group was 92.91% (891/959), which was higher than 58.64% (577/984) in the control group, and the difference was statistically significant ( χ2=15.28, P<0.05). Conclusions:The implementation of BTS quality improvement model is helpful to improve the nursing quality of patients with indwelling NCC dialysis and improve the patient outcomes.
4.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.
5.Study on the Predictive Value of Serum VEGFR-1 and MIP-3α Level Expression in Endometriosis Patients on Postoperative Recurrence
Xiaohua SUN ; Yulin SONG ; Hong LI
Journal of Modern Laboratory Medicine 2024;39(4):170-174
Objective To investigate the expression of serum vascular endothelial growth factor receptor-1(VEGFR-1)and macrophage inflammatory protein-3α(MIP-3α)in patients with endometriosis(EMs)patients and the predictive value of their combined detection for postoperative recurrence in EMs patients.Methods A total of 114 patients with EMs who underwent laparoscopic surgery in Maternal and Child Care Hospital of Qinhuangdao from April 2019 to June 2021 were selected as the observation group,while 114 healthy medical checkups of women of gestational age who had their medical checkups in the hospital were selected as the control group during the same period.Enzyme-linked immunosorbent assay(ELISA)was used to determine the serum levels of VEGFR-1 and MIP-3α in the patients.According to the postoperative period of 2 years,they were divided into the recurrence group(n=78)and the non-recurrence group(n=36).Multivariate logistic regression analysis was used to analyze influencing factors of postoperative recurrence in patients with EMs.The predictive value of the combined serum VEGFR-1 and MIP-3α test for postoperative recurrence in patients with EMs was analyzed by using the receiver operating characteristic curve(ROC)curve.Results Compared with the control group,the levels of VEGFR-1 in the observation group(116.25±48.57 pg/ml vs 92.43±25.37 pg/ml)and MIP-3α(19.25±5.24 pg/ml vs 13.67±4.28 pg/ml)were elevated,and the differences were significant(t=4.641,8.806,all P<0.05).The level of VEGFR-1(104.22±5.78 pg/ml,118.60±6.56 pg/ml,138.55±7.85 pg/ml)and the level of MIP-3α(15.37±1.15 pg/ml,19.28±2.12 pg/ml,25.42±2.56 pg/ml)in mild,moderate and severe EMs patients were increased successively,and the differences were significant(F=147.757,133.654,all P<0.001).The percentage of the presence of palpable nodules in the posterior fornix,r-AFS staging(stage Ⅲ~Ⅳ)in the recurrence group was greater than those in the non-recurrence group(x2=15.139,10.310,all P<0.05),and the percentage of the recurrence group with postoperative medication for 6 months or more was lower than that of the non-recurrence group(x2=15.016,P<0.001),and the differences were statistically significant,respectively.Multivariate logistic regression analysis showed that serum VEGFR-1,MIP-3α,the presence of palpable nodules in the posterior fornix and r-AFS staging were risk factors for postoperative recurrence of EMs(all P<0.05),while postoperative medication for 6 months or more was a protective factor(P<0.05).The ROC curves showed that the area under the curve(AUC)of the combination of serum VEGFR-1 and MIP-3αpredicted postoperative EMs for postoperative recurrence was the largest(0.929),and its sensitivity and specificity were 85.90%and 86.11%,respectively.Conclusion VEGFR-1 and MIP-3α expression was elevated in the serum of patients with EMs.The efficacy of their combined detection in predicting recurrence after EMs could be better.
6.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.
7.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.
8.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.
9.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.
10.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.

Result Analysis
Print
Save
E-mail