1.Prokaryotic expression,purification of coxsackievirus A16 Vp1 protein and preparation of rabbit polyclonal antibodies
Jianing WANG ; Yongjuan LIU ; Kaikai RAN ; Binlian SUN ; Yingying SHI
Journal of Jilin University(Medicine Edition) 2025;51(6):1717-1727
Objective:To construct the prokaryotic expression vector of Coxsackievirus A16(CA16)viral protein 1(Vp1),express it in Escherichia coli(E.coli)BL21,purify the protein,prepare rabbit polyclonal antibodies,and identify the immunoreactivity of the antibodies.Methods:Bioinformatics online tools were used to predict the amino acid composition,conserved domains,secondary and tertiary structures of the Vp1 protein.The Vp1 gene of CA16 was amplified and cloned into the prokaryotic expression vector pET28a(+).The pET28a-Vp1 was transformed into E.coli BL21,and the expression was induced using isopropyl β-D-thiogalactopyranoside(IPTG).Western blotting method was used to identify the induced expression of Vp1 protein,and the induction time and temperature were optimized to improve the expression efficiency.Two female SPF New Zealand white rabbits were taken,and the purified recombinant protein Vp1 was used to immunize the rabbits to prepare rabbit anti-Vp1 protein polyclonal antibodies.The antibody titer was determined by ELISA method,and the immunoreactivity of the antibodies was identified by Western blotting method.Results:The bioinformatics analysis results showed that the Vp1 gene encoded 297 amino acids,with a relative molecular mass of 33 046.39 and an isoelectric point of 8.32,belonging to a hydrophilic protein;in the secondary structure,α-helix accounted for 15.15%,random coil accounted for 67.68%,and extended strand accounted for 17.17%.Sequencing of the recombinant plasmid pET28a-Vp1 confirmed that the pET28a-Vp1 plasmid was correctly constructed.The Western blotting results showed that the target protein was expressed in the IPTG induction group at a relative molecular mass of 33 000,and the target protein was mainly expressed in inclusion bodies.The optimal induction conditions for protein expression were IPTG concentration of 0.4 mmol·L-1,temperature of 16℃,and induction time of 20 h.The ELISA assay results showed that the titer of the rabbit polyclonal antibody was 1:1 024 000.The Western blotting results showed that the Vp1 rabbit polyclonal antibody could bind to the viral Vp1 protein in the CA16-infected cells.Conclusion:The polyclonal antibody against CA16 Vp1 is successfully prepared,and this antibody has significant binding characteristics to CA16 Vp1,which can be used for the diagnosis of enterovirus infection and the development of treatment methods.
2.Application of integrated Chinese and western medicine nursing based on the individual and family self-management in post-stroke urinary incontinence patients
Yongjuan YAO ; Peng WANG ; Shasha NAN ; Miaomiao FAN ; Zhenlan HONG
Chinese Journal of Practical Nursing 2024;40(13):961-969
Objective:To explore the application effect of integrated Chinese and Western medicine nursing based on the individual and family self-management theory (IFSMT) in patients with urinary incontinence after stroke, and provide reference for clinical nurses to effectively rehabilitate these patients.Methods:This was a quasi-experimental study. A total of 136 patients with urinary incontinence after stroke who were hospitalized in the Department of Encephalopathy of Shanxi Acupuncture and Moxibology Hospital from February 2022 to May 2023 were selected as the study objects by convenience sampling method. The subjects were divided into the control group and the experimental group with 68 cases in each group by random number table method. The control group received routine rehabilitation nursing, and the experimental group received integrated Chinese and western medicine nursing based on IFSMT on the basis of routine rehabilitation nursing. Urinary status, activities of daily living and quality of life of 2 groups were compared.Results:Ultimately, there were 68 cases were admitted in each group. In the control group, male 45 cases, female 23 cases, aged (62.35 ± 4.94) years old. In the experimental group, male 49 cases, female 19 cases, aged (61.94 ± 5.02) years old. There was no statistical significance in urinary status, activities of daily living and quality of life scores between the 2 groups before the intervention (all P>0.05). After the intervention, the average daily urination frequency of experimental group was (7.94 ± 1.08) times, which was lower than that of control group (9.88 ± 1.09) times; the average daily frequency of incontinence in the experimental group was (3.63 ± 1.65) times, lower than (5.03 ± 3.35)times in the control group; the average daily urine volume of experimental group was (107.34 ± 4.15) ml, higher than (89.62 ± 19.71) ml of control group, and the differences were statistically significant ( t=10.46,3.09, -7.23, all P<0.05). The scores of activities of daily living in the experimental group was (60.44 ± 3.65) points, which were higher than (46.24 ± 3.29) points in the control group; the Incontinence Quality of Life Questionnaire score of experimental group was (80.38 ± 4.65) points, which was higher than (64.62 ± 5.62) points of control group; Insomnia Severity Index-Short Form score of the experimental group was (10.94 ± 5.17) points, which was lower than (12.85 ± 4.30) points of the control group, and the differences were statistically significant ( t=23.85, -17.86, 2.34, all P<0.05). Conclusions:Integrated Chinese and western medicine nursing intervention based on IFSMT has a positive effect on patients with urinary incontinence after stroke, and can significantly improve patients' urination and daily living activities, thus improving patients' quality of life.
3.Clinicopathological characteristics of gangliogliomas with anaplastic morphology
Linai GUO ; Leiming WANG ; Yongjuan FU ; Tao LUO ; Xiaotong FAN ; Lihong ZHAO ; Xiaohong YAO ; Yueshan PIAO
Chinese Journal of Pathology 2024;53(6):585-591
Objective:To investigate the clinical, radiological, and pathological features of anaplastic gangliogliomas (AGGs) and to determine whether these tumors represent a distinct entity.Methods:Consecutive 667 cases of ganglioglioma (GG) diagnosed at the Xuanwu Hospital, Capital Medical University, Beijing, China between January 2015 and July 2023 were screened. Among these cases, 9 pathologically confirmed AGG cases were identified. Their clinical, radiological, treatment, and outcome data were analyzed retrospectively. Most of the tumor samples were subject to next-generation sequencing, while a subset of them were subject to DNA methylation profiling.Results:Among the 9 patients, there were five males and four females, with a median age of 8 years. Epileptic seizures (5/9) were the most frequently presented symptom. Radiological examinations showed three types of radiological manifestations: four cases showed abnormal MRI signals with no significant mass effects and mild enhancement; two cases demonstrated a mixed solid-cystic density lesion with peritumoral edema, which showed significant heterogeneous enhancement and obvious mass effects, and one case displayed cystic cavity formation with nodules on MRI, which showed evident enhancements. All cases exhibited mutations that were predicted to activate the MAP kinase signaling pathway, including seven with BRAF p.V600E mutation and two with NF1 mutation. Five AGGs with mutations involving the MAP kinase signaling pathway also had concurrent mutations, including three with CDKN2A homozygous deletion, one with a TERT promoter mutation, one with a H3F3A mutation, and one with a PTEN mutation.Conclusions:AGG exhibits a distinct spectrum of pathology, genetic mutations and clinical behaviors, differing from GG. Given these characteristics suggest that AGG may be a distinct tumor type, further expansion of the case series is needed. Therefore, a comprehensive integration of clinical, histological, and molecular analyses is required to correctly diagnose AGG. It will also help guide treatments and prognostication.
4.Application of enhanced recovery after surgery in perioperative management of complicated appendicitis in children
Huazhe WU ; Yongjuan WEI ; Qiuju WANG ; Jiarong LIU
China Modern Doctor 2024;62(13):11-14
Objective To explore the effect of enhanced recovery after surgery(ERAS)in perioperative period of complicated appendicitis in children.Methods A total of 248 children with complicated appendicitis who underwent laparoscopic surgery in Quanzhou Children's Hospital from January 2020 to January 2023 were selected and divided into control group and ERAS group according to random number table method,with 124 cases in each group.Both groups of patients underwent laparoscopic appendectomy.The control group received traditional treatment during perioperative period,while the ERAS group received treatment based on the concept of ERAS.The first postoperative exhaust/defecation time,hospital stay,visual analogue scale(VAS)score,postoperative complications and hospitalization satisfaction were compared between two groups.Results The time of first postoperative exhaust/defecation and hospital stay in ERAS group were significantly shorter than those in control group,total incidence of postoperative complications and postoperative VAS score were significantly lower than those in control group,and hospitalization satisfaction was significantly better than that in control group(P<0.05).Conclusion ERAS implementation during perioperative period of complicated appendicitis in children can promote early postoperative recovery,reduce postoperative pain,reduce the incidence of complications,and improve hospitalization satisfaction,which is worthy of clinical promotion and application.
5.Functional annotation map of natural compounds in traditional Chinese medicines library: TCMs with myocardial protection as a case.
Xudong XING ; Mengru SUN ; Zifan GUO ; Yongjuan ZHAO ; Yuru CAI ; Ping ZHOU ; Huiying WANG ; Wen GAO ; Ping LI ; Hua YANG
Acta Pharmaceutica Sinica B 2023;13(9):3802-3816
The chemical complexity of traditional Chinese medicines (TCMs) makes the active and functional annotation of natural compounds challenging. Herein, we developed the TCMs-Compounds Functional Annotation platform (TCMs-CFA) for large-scale predicting active compounds with potential mechanisms from TCM complex system, without isolating and activity testing every single compound one by one. The platform was established based on the integration of TCMs knowledge base, chemome profiling, and high-content imaging. It mainly included: (1) selection of herbal drugs of target based on TCMs knowledge base; (2) chemome profiling of TCMs extract library by LC‒MS; (3) cytological profiling of TCMs extract library by high-content cell-based imaging; (4) active compounds discovery by combining each mass signal and multi-parametric cell phenotypes; (5) construction of functional annotation map for predicting the potential mechanisms of lead compounds. In this stud TCMs with myocardial protection were applied as a case study, and validated for the feasibility and utility of the platform. Seven frequently used herbal drugs (Ginseng, etc.) were screened from 100,000 TCMs formulas for myocardial protection and subsequently prepared as a library of 700 extracts. By using TCMs-CFA platform, 81 lead compounds, including 10 novel bioactive ones, were quickly identified by correlating 8089 mass signals with 170,100 cytological parameters from an extract library. The TCMs-CFA platform described a new evidence-led tool for the rapid discovery process by data mining strategies, which is valuable for novel lead compounds from TCMs. All computations are done through Python and are publicly available on GitHub.
6.Research advances in the regulation of epithelial-mesenchymal transition and targeted therapy for liver fibrosis
Yongjuan WANG ; Xiaoli XIE ; Huiqing JIANG
Journal of Clinical Hepatology 2021;37(1):165-168
The pathological basis of liver fibrosis is the deposition of extracellular matrix (ECM), and myofibroblasts are the main source of ECM. Epithelial-mesenchymal transition (EMT) is one of production mechanisms of myofibroblasts. At present, a large number of studies have shown that intervention of key EMT molecules and signaling pathways as targets can reduce liver fibrosis. Based on literature review, this article summarizes the signaling pathways associated with EMT, important regulatory molecules, and drugs targeting EMT in the treatment of liver fibrosis, so as to provide new ideas for the treatment of liver fibrosis.
7.Association of genetic variants in renalase with blood pressure responses to salt and potassium intake
Yang WANG ; Yue SUN ; Guilin HU ; Ting ZOU ; Xiaoyu ZHANG ; Mingfei DU ; Haowei ZHOU ; Hao JIA ; Dan WANG ; Jie ZHANG ; Chen CHEN ; Jiawen HU ; Qiong MA ; Yue YUAN ; Yueyuan LIAO ; Keke WANG ; Yu YAN ; Xi ZHANG ; Zejiaxin NIU ; Yongjuan GUAN ; Ruichen YAN ; Ke GAO ; Min LI ; Jianjun MU
Journal of Xi'an Jiaotong University(Medical Sciences) 2021;42(3):392-397
【Objective】 Based on our previously established salt-sensitive hypertension cohort, we conducted chronic salt loading and potassium supplementation interventions, aiming to examine the association between genetic variants in renalase and blood pressure (BP) responses to dietary interventions of salt and potassium intake. 【Methods】 In 2004, 514 subjects from 126 families were recruited in Shaanxi Province to establish the salt-sensitive hypertension study cohort. Among them, 334 non-parent subjects were selected and sequentially maintained on a low-salt diet for 7 days, then a high-salt diet for 7 days and a high-salt diet with potassium supplementation for another 7 days. Ten single nucleotide polymorphisms (SNPs) in the renalase gene were genotyped on the MassARRAY platform. 【Results】 SNP rs2576178 of the renalasegene was significantly associated with systolic BP (SBP) and mean arterial pressure (MAP) responses to low-salt intervention (SBP: β=-2.730, P<0.05; MAP: β=-1.718, P<0.05). In addition, SNP rs12356177 was significantly associated with diastolic BP response to low-salt diet (β=-1.608, P<0.05). However, we did not find any association for the renalase SNPs with BP response to high-salt diet with potassium supplementation reached nominal statistical significance. 【Conclusion】 Genetic variants in renalase gene are significantly associated with BP response to low-salt diet, suggesting that renalase may be mechanistically involved in BP salt-sensitivity.
8.Clinicopathological features of diffuse leptomeningeal glioneuronal tumor
Jinhua TU ; Leiming WANG ; Li LIU ; Haiwei HAN ; Yongjuan FU ; Yueshan PIAO ; Dehong LU ; Lianghong TENG
Chinese Journal of Pathology 2021;50(8):876-881
Objective:To investigate the clinicopathological features, diagnosis and prognosis of diffuse leptomeningeal glioneuronal tumor (DLGNT).Methods:Five cases of DLGNT diagnosed from January 2016 to January 2020 were collected from Xuanwu Hospital, Capital Medical University. The clinical features, histopathologic characteristics, immunohistochemical and molecular genetic findings and prognosis were analyzed and the relevant literature was reviewed.Results:The five patients (two males and three females) were aged 2 to 52 years (median 11 years), and had history of increased intracranial pressure (headache and vomiting) or limb weakness. Three of them were younger than 16 years of age. The imaging studies showed diffuse intracranial and intraspinal nodular leptomeningeal thickening and enhancement, with or without parenchymal involvement. At times there were associated small cyst-like lesions. Imaging interpretations were inflammatory lesions in three cases and space occupying lesions in two. Microscopically, in three cases the tumors showed low to moderate cellularity, consisting of relatively monomorphous oligodendrocyte-like cells arranged in small nests or diffusely distribution. No mitosis and necrosis were observed. In two cases there were increased cellularity with a diffuse honeycomb pattern. The tumor showed mild to moderate polymorphism with hyperchromatic nuclei. Mitosis, endothelial vascular proliferation and glomeruloid vessels were seen. Necrosis was absent. The tumor cells in all five cases were positive for synaptophysin,Olig2 and negative for IDH1 and H3 K27M. GFAP was focally positive in four cases and only one case expressed NeuN partly. The Ki-67 labeling index was 1%-35%. BRAF fusion was detected in four cases. Genetic analysis showed solitary 1p deletion in two cases (2/5), while all cases were negative for 1p/19q co-deletion (0/5). The five patients were followed up for 13 to 28 months (median 15 month). One patient died after 27 months. There was no evidence of tumor progression in the remaining four patients.Conclusions:DLGNT is rare and easily confused with other central nervous system tumors and inflammatory lesions. Therefore, the diagnosis of DLGNT should be made based on comprehensive information including imaging, morphologic and corresponding immunohistochemical examinations and molecular genetics to avoid misdiagnosis and delay in management.
9.A comparative study on the clinical effects between laparoscopic ballon dilation and traditional open reduction in treatment of intussusception in children
Chengji ZHAO ; Yongjuan ZENG ; Zhongfu MA ; Binde LI ; Gang LI ; Wenyun WANG ; Jike HU
Chinese Journal of Applied Clinical Pediatrics 2020;35(11):860-863
Objective:To investigate and compare the therapeutic effect of Foley catheter balloon dilation and the traditional open reduction in the treatment of intussusception in children.Methods:A total of 68 children with intussusception treated by Foley catheter balloon dilatation by laparoscopic surgery or traditional open reduction in Department of Pediatric Surgery, the Second Hospital of Lanzhou University from August 2015 to July 2019 were retrospectively analyzed.Among them, there were 32 cases in the Foley catheter group and 36 cases in the traditional laparotomy group.Foley catheter group were treated with laparoscopic Foley catheter balloon dilatation, while the traditional open group were treated with traditional open surgery which was performed with finger dilatation.The results of surgical treatment, postoperative recovery, short-term complications, patient satisfaction and long-term complications were compared between the two groups.Results:Among the indexes of surgical treatment effect, the operation time[(0.4±1.1) h], intraoperative bleeding volume[(10.2±3.4) mL], incision size[(0.5±0.4) cm] and incidence of the rupture of intestine[6.3%(2/32 cases)] in the Foley catheter group were significantly lower than those in the traditional open group[(1.3±2.9) h, (40.5±2.1) mL, (5.1±0.7) cm, 30.6%(11/36 cases)], and the differences were statistically significant (all P< 0.05). Among the indexes of postoperative recovery and short-term complications, recovery time of gastrointestinal function[(1.2±3.1) d], length of hospital stay[(6.7±1.8) d], incidence of incision infection[9.4%(3/32 cases)]and incidence of incisional hernia(0) in the Foley catheter group were significantly lower than those in the traditional open group[(3.3±6.4) d, (7.3±0.9) d, 36.1%(13/36 cases), 16.7%(6/36 cases)], and the differences were statistically significant (all P<0.05). Among the indexes of family satisfaction and long-term complications, the score of family satisfaction in the Foley catheter group [(8.7±1.2) scores]was significantly higher than that in the traditional open group[(6.6±3.1) scores], and the incidence of adhesive intestinal obst-ructionin the Foley catheter group (0)was significantly lower than that in the traditional open group[0 vs.19.4%(7/36例)], and the differences were statistically significant(all P<0.05). Conclusions:Compared with traditional open reduction in the treatment of intussusception in children, Foley catheter balloon dilation has the advantages of short operation time, safe operation, low incidence of intestinal injury, less bleeding, and so on, and also has the advantages of small incision, fast recovery, short hospitalization time, high satisfaction of parents.In addition, the Foley catheter balloon dilation has a lower incidence of incisional infection, incisional hernia, postoperative intestinal adhesion and other complications.
10.Association between neuroimmunological indicators and autism spectrum disorders
WANG Lin, YANG Xiaolei, MA Yongjuan, LIU Xiaocui, ZOU Mingyang, SUN Caihong, WU Lijie
Chinese Journal of School Health 2019;40(4):499-502
Objective:
To explore the relationship among osteopontin(OPN), Interleukin-17A(IL-17A), anti-MBP auto-antibody and autism spectrum disorder(ASD), and to provide the theoretical basis for the etiology and pathogenesis of ASD.
Methods:
Forty autistic children and forty matched healthy children were enrolled in this case-control study. The levels of OPN, IL-17A, anti-MBP autoantibody in serum were measured by enzyme-linked immunosorbent assay (ELISA). The associations between those metabolic levels and the severity and intelligence of ASD children were performed by Pearson or Spearman correlation.
Results:
Children with ASD had higher serum levels of OPN, IL-17A [(296.89±162.95),0.93] pg/mL compared to healthy control[(217.98±113.39), 0.62] pg/mL(P<0.05). Serum OPN, IL-17A, and anti-MBP auto-antibody levels in ASD group were not correlated with the scores of ABC, CARS, and PPVT(P>0.05). However, anti-MBP auto-antibodies level in children with ASD were positively correlated with OPN and IL-17A levels, respectively(r=0.35, 0.34, P<0.05).
Conclusion
It was obvious that the ASD children were found with neuroimmunologic abnormality, and the underlying mechanism needs to be further explored.


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