1.Finite element analysis of various root shield thicknesses in maxillary central incisor socket-shield technique
Guangneng CHEN ; Siyang LUO ; Mei WANG ; Bin YE ; Jiawen CHEN ; Yin LIU ; Yuwen ZUO ; Xianyu HE ; Jiajin SHEN ; Minxian MA
Chinese Journal of Tissue Engineering Research 2025;29(10):2052-2060
BACKGROUND:Socket-shield technique can effectively maintain labial soft and hard tissues,but the incidence of postoperative complications such as exposure and displacement of root shield is relatively high.It is speculated that the root shield may be exposed and displaced due to excessive load after long-term function of dental implants. OBJECTIVE:Through three-dimensional finite element analysis,we aim to study the influence of varying root shield thicknesses on the stress distribution,equivalent stress peaks,and displacement in the root shield,periodontal ligaments,implant,and surrounding alveolar bone under normal occlusal loading.We also attempt to analyze the correlation between the thickness of the root shield and occurrence of mechanical events such as root shield exposure,displacement,and fracture. METHODS:Cone-beam CT data of a patient who met the indication standard of socket-shield technique for maxillary central incisor were retrieved from database.Reverse engineering techniques were used to build models of the maxillary bone and root shield,while forward engineering was used to create models for the implant components based on their parameters.Models depicting various root shield thicknesses(0.5,1.0,1.5,and 2.0 mm)were created using Solidworks 2022 software.ANSYS Workbench 2021 software was then used to simulate and analyze the effects of varying root shield thicknesses on stress distribution,equivalent stress peaks,and displacement of the root shields,periodontal ligaments,implants,and surrounding alveolar bone under normal occlusion. RESULTS AND CONCLUSION:(1)In all root shield models,the stress was concentrated on the palatal cervical side,both sides of the edges and the lower edge of the labial side.As the thickness of the root shield increased,the equivalent stress peak and displacement showed a decreasing trend.The 0.5 mm thickness model produced a stress concentration of 176.20 MPa,which exceeded the yield strength(150 MPa)of tooth tissue.(2)The periodontal ligament stress in each group was concentrated in the neck margin and upper region.With the increase of root shield thickness,the equivalent stress peak and displacement of periodontal ligament showed a decreasing trend.(3)Implant stress in all models was concentrated in the neck of the implant and the joint of the implant-repair abutment,and the labial side was more concentrated than the palatal side.With the increase of root shield thickness,the equivalent stress peak of the implant in the model showed an increasing trend.(4)In each group of models,stress of cortical bone concentrated around the neck of the implant and the periphery of the root shield,and the labial side was more concentrated than the palatal side.With the increase of the thickness of the root shield,the equivalent stress peak around the root shield decreased;the peak value of the equivalent stress of the bone around the neck of the implant showed an increasing trend.In the model,the stress of cancellous bone was mainly concentrated around the neck of the lip of the implant,the top of the thread,the root tip and the lower margin of the root shield,and the labial side was more concentrated than the palatal side.With the increase of the thickness of the root shield,the peak value of the equivalent stress of the bone around the root shield in the model showed a decreasing trend.The minimum principal stress of cortical bone in each group of models was concentrated around the neck of the implant,exhibiting a fan-shaped distribution.As the thickness of the root shield increased,the minimum principal stress of cortical bone showed an increasing trend.(5)These results indicate that different thicknesses of the root shield have different biomechanical effects.The root shield with a thickness of 0.5 mm is easy to fracture.For patients with sufficient bone width,the root shield with a thickness of 2.0 mm is an option to reduce the risk of complications such as root shield exposure,fracture,and displacement.Meanwhile,it should be taken into account to protect the periodontal ligament in the preparation process,and rounding treatments ought to be carried out on both sides and the lower edge of the root shield.
2.Expression and pathological diagnostic value of extracellular matrix metalloproteinase inducer,ma-trix metalloproteinase-9 and lysine demethylase 6B in invasive breast cancer
Huang JIANG ; Lihua ZHENG ; Xiaoyan XU ; Jianjun WANG ; Xianwei XU ; Na WANG ; Chenju XING ; Xianyu LU
Journal of Xinxiang Medical College 2024;41(2):143-150
Objective To investigate the expressions of extracellular matrix metalloproteinase inducer(EMMPRIN),matrix metalloproteinase-9(MMP-9),lysine demethylase 6B(KDM6B)proteins and their correlation with clinicopathologic features in invasive breast cancer,and analyze the correlation among the three proteins and their value in pathological diagnosis of invasive breast cancer.Methods The surgical biopsy specimens of 124 patients with invasive breast cancer who were admitted to the Department of Pathology,the Fifth Clinical Medical College of Henan University of Chinese Medicine/People's Hospital of Zhengzhou from January 2014 to December 2017 were selected as research subjects,and 20 low-grade intraductal carcinoma tissue specimens,27 high-grade intraductal carcinoma tissue specimens,and 22 adjacent tissue specimens>1 cm away from the invasive breast cancer were selected as controls.The expressions of EMMPRIN,MMP-9 and KDM6B proteins in cancer-adjacent tissues,low-grade intraductal carcinoma tissues,high-grade intraductal carcinoma tissues and invasive breast cancer tissues were detected by immunohistochemistry.The relationship between the relative expressions of EMMPRIN,MMP-9 and KDM6B proteins and clinicopathologic features of invasive breast cancer was analyzed,Spearman correlation analysis was used to evaluate the correlation of EMMPRIN,MMP-9 and KDM6B proteins in breast cancer tissues,and receiver operating characteristic(ROC)curve was adopted to evaluate the diagnostic value of EMMPRIN,MMP-9 and KDM6B for invasive breast cancer.Results The relative expressions of EMMPRIN and MMP-9 proteins in high-grade intraductal carcinoma and invasive breast cancer tissues were significantly higher those in cancer-adjacent tissues and low-grade intraductal carcinoma tissues,and the relative expression of KDM6B protein was significantly lower than those in cancer-adjacent tissues and low-grade intraductal carcinoma tissues(P<0.05);the relative expressions of EMMPRIN and MMP-9 proteins in invasive breast cancer tissues were significantly higher those in high-grade intraductal carcinoma tissues,and the relative expression of KDM6B protein was significantly lower than that in high-grade intraductal carcinoma tissues(P<0.05);there was no statistically significant difference in the relative expressions of EMMPRIN,MMP-9 and KDM6B proteins between cancer-adjacent tissues and low-grade intraductal carcinoma tissues(P>0.05).The relative expressions of EMMPRIN and KDM6B proteins were not related to the age,tumor location and tumor diameter of patients with invasive breast cancer(P>0.05),and the relative expression of MMP-9 protein was not related to the age and tumor location of patients with invasive breast cancer(P>0.05).Relative expressions of EMMPRIN,MMP-9 and KDM6B proteins were correlated with WHO grading,lymph node metastasis,and tumor,node and metastasis(TNM)staging of invasive breast cancer(P<0.05),and the relative expression of MMP-9 protein was correlated with the tumor diameter(P<0.05).In the WHO grades Ⅰ,Ⅱ,and Ⅲ of invasive breast cancer,the relative expressions of EMMPRIN and MMP-9 proteins increased sequentially,while the relative expression of KDM6B protein decreased sequentially(P<0.05);the relative expressions of EMMPRIN and MMP-9 proteins in the lymph node metastasis group were significantly higher than those in the non-lymph node metastasis group,and the relative expression of KDM6B protein was significantly lower than that in the non-lymph node metastasis group(P<0.05);the relative expressions of EMMPRIN and MMP-9 proteins in TNM stages Ⅲ-Ⅳ were significantly higher than those in stages Ⅰ-Ⅱ(P<0.05),while the relative expression of KDM6B protein was significantly lower than that in stages Ⅰ-Ⅱ(P<0.05).In the group of invasive breast cancer with diameter≤2 cm,2 to 5 cm,and>5 cm,the relative expression of MMP-9 protein increased sequentially(P<0.05).Spearman correlation analysis showed that the expression of EMMPRIN was positively correlated with MMP-9 protein in invasive breast cancer tissues(r=0.990,P=0.000),the expression of EMMPRIN was negatively correlated with KDM6B protein(r=-0.606,P=0.000),and the expression of MMP-9 was negatively correlated with KDM6B protein(r=-0.612,P=0.000).ROC curve analysis showed that the area under the curve(AUC)of EMMPRIN protein for diagnosing invasive breast cancer was 0.875[95%confidence interval(CI):0.823-0.926,P<0.05],with an optimal threshold of 10.043,sensitivity of 79.0%,and specificity of 76.8%;the AUC of MMP-9 protein in diagnosing invasive breast cancer was 0.863(95%CI:0.808-0.917,P<0.05),with an optimal threshold of 10.070,sensitivity of 74.2%,and specificity of 76.8%;the AUC of KDM6B protein in diagnosing invasive breast cancer was 0.267(95%CI:0.196-0.338,P<0.05),with an optimal threshold of 11.003,sensitivity of 71.0%,and specificity of 98.6%.Conclusion EMMPRIN,MMP-9 and KDM6B are related to the occurrence and development of invasive breast cancer.Detection of the expressions of EMMPRIN,MMP-9 and KDM6B is helpful to the pathological diagnosis of invasive breast cancer and clinical judgment of invasion and metastasis of breast cancer.
3.Mechanism of Treatment of Hepatocellular Carcinoma with Traditional Chinese Medicine Based on Epigenetic Regulation: A Review
Xianyu XU ; Yongping ZHU ; Yanqing LIU ; Liwei GU ; Junzhe ZHANG ; Shengnan SHEN ; Jigang WANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(23):281-291
Hepatocellular carcinoma (HCC) is the sixth most common cancer in the world. In recent years, the clinical early diagnosis and treatment protocols of HCC have been improved, whereas the prognosis of patients is still not satisfactory, which is due to the fact that the mechanism of HCC development has not been fully elucidated. Therefore, it is of great significance to explore the molecular mechanisms and key regulatory links of hepatocellular carcinoma development to further improve the diagnosis and treatment of HCC in China. Epigenetics has become a research hotspot because of its reversibility and easy regulation. According to relevant studies, HCC involves the accumulation of multiple genetic and epigenetic changes during the initiation, promotion, and progression stages. HCC is categorized as infantile malnutrition with accumulation, hypochondriac pain, tympan ites, and abdominal mass in traditional Chinese medicine (TCM). In the treatment of HCC, TCM with low toxicity, multi-targets, and multi-mechanisms can inhibit tumor growth, alleviate the clinical symptoms, and enhance the quality of life of the patients. Chinese medicines and their active ingredients exert anti-HCC effects through epigenetic regulation of DNA methylation, histone modification, and non-coding RNA. Abnormal gene expression due to epigenetic regulation disorders is involved in all stages of HCC development. There are few studies on epigenetic regulation in TCM treatment of HCC, and there is still much room for development in basic and clinical trials. This paper reviews the mechanism of epigenetic regulation in HCC and summarizes the experimental results of TCM research on the related mechanism, with a view to providing a theoretical basis for future research on the mechanism of HCC development and clinical diagnosis and treatment of hepatocellular carcinoma with TCM.
4.Analysis of clinical implementation of nursing group standard for oxygen therapy in adult patients
Xiaojiu QI ; Huiting WANG ; Yu XU ; Zheng HUANG ; Zhixia ZHANG ; Hongwen MA ; Yunyan XIANYU ; Su ZHANG
Chinese Journal of Nursing 2024;59(14):1726-1731
Objective To understand the implementation of the nursing group standards for oxygen inhalation therapy in clinical practice,and to provide a reference for improving the nursing practice of oxygen therapy.Methods A convenience sampling method was used to investigate nurses from 902 hospitals in 24 provinces and municipalities directly under the central government using a self-designed questionnaire from December 15th,2022,to January 14th,2023.The content of questionnaire included whether they had implemented the recommendations of the oxygen therapy standards,the knowledge of safety related to oxygen therapy,and the components of oxygen therapy prescriptions,the indications used for patients receiving oxygen therapy and practice status of oxygen therapy.Results A total of 10481 questionnaires were returned,of which 10447 were valid,with a valid questionnaire recovery rate of 99.68%.63.14%of the nurses indicated that the hospital had organized training on oxygen therapy standards.Only 47.82%of nurses know the correct use of the Venturi mask.41.90%of nurses could indicate the correct indicator of flow adjustment.31.88%of the nurses stated that they will adjust the oxygen flow rate based on the oxygenation status of carbon dioxide storage patients.Only 19.56%of nurses indicated that humidification is applied in oxygen therapy based on the oxygen flow and duration.Conclusion Even though nurses had received training related to oxygen therapy standards,the level of knowledge of oxygen therapy standards was still low;therefore continuous systematic training was needed,and the implementation of the content of oxygen therapy standards needed to be further standardized.Healthcare institutions would focus on organizing systematic training and maintaining the training effect,enhancing infrastructure and providing support for implementation.Recommendation to the nursing administration is to explore how to comprehensively and continuously implementing the oxygen therapy nursing standards with the ultimate goal of providing patients safer and more accurate oxygen therapy.
5.The clinical outcomes of endovenous microwave ablation and radiofrequency ablation for great saphenous varicose veins
Zhiyong CHEN ; Long WANG ; Huan OUYANG ; Xianyu HU ; Yuan HONG ; Bo LIU ; Yi LIU ; Xu ZHANG ; Binshan ZHA
Chinese Journal of General Surgery 2023;38(10):765-771
Objective:To compare the efficacy of intraluminal microwave ablation with radiofrequency ablation in the treatment of varicose veins of the lower extremities.Methods:The clinical data of 520 patients (522 affected limbs) who underwent lower extremity varicose vein surgery at the Department of Vascular Surgery of the First Affiliated Hospital of Anhui Medical University from Jun 2021 to Sep 2022 were collected. Patients were divided into endovenous microwave ablation group (EMWA group, n = 201) and radiofrequency ablation group (RFA group, n = 321). Follow-up was performed at 1 week, 1 , 6 and 12 months after surgery. The primary efficacy endpoint was the occlusion rate of the treated segment vein, the primary safety endpoint was the incidence of surgery-related and/or device-related complications, and the secondary endpoints were the venous clinical severity score (VCSS) and chronic venous insufficiency quality of life questionnaire (CIVIQ) scores at follow-up. Results:The technique success rate and the occlusion rate of the affected segment vein was 100% in both groups evaluated one week after surgery; Six and 12 months after surgery, the occlusion rate in the RFA group was 98.9%, and that in the EMWA group was 99.3% and respectively 97.8%, 97.2% ,without statically significant difference.During the follow-up period, there were no cases of reoperation due to vein recanalization. no serious events such as deep vein thrombosis, pulmonary embolism or death occurred in either group. The incidence of adverse events (induration, ecchymosis, skin burn, incision infection, limb numbness, hematoma, thrombotic superficial phlebitis, endovenous heat induced thrombosis, etc.) in both groups was compared, and the difference was not statistically significant. VCSS and CIVIQ scores improved significantly in both groups at 1 ,6 and 12 months after treatment, and the difference was statistically significant(all P < 0.01). Conclusion:EMWA and RFA have the advantages of simple operation, good clinical efficacy and high degree of improvement in quality of life.
6.Isolation of High Purity Mouse Mesenchymal Stem Cells through Depleting Macrophages Using Liposomal Clodronate
Ju Han SONG ; Jung-Woo KIM ; Mi Nam LEE ; Sin-Hye OH ; Xianyu PIAO ; Zhao WANG ; Seung-Hee KWON ; Ok-Su KIM ; Jeong-Tae KOH
Tissue Engineering and Regenerative Medicine 2022;19(3):565-575
BACKGROUND:
The use of mouse bone marrow mesenchymal stem cells (mBMSCs) represents a promising strategy for performing preclinical studies in the field of cell-based regenerative medicine; however, mBMSCs obtained via conventional isolation methods have two drawbacks, i.e., (i) they are heterogeneous due to frequent macrophage contamination, and (ii) they require long-term culturing for expansion.
METHODS:
In the present study, we report a novel strategy to generate highly pure mBMSCs using liposomal clodronate.This approach is based on the properties of the two cell populations, i.e., BMSCs (to adhere to the plasticware in culture dishes) and macrophages (to phagocytose liposomes).
RESULTS:
Liposomal clodronate added during the first passage of whole bone marrow culture was selectively engulfed by macrophages in the heterogeneous cell population, resulting in their effective elimination without affecting the MSCs.This method allowed the generation of numerous high-purity Sca-1 + CD44 + F4/80 - mBMSCs ([ 95%) with just one passaging. Comparative studies with mBMSCs obtained using conventional methods revealed that the mBMSCs obtained in the present study had remarkably improved experimental utilities, as demonstrated by in vitro multilineage differentiation and in vivo ectopic bone formation assays.
CONCLUSION
Our newly developed method, which enables the isolation of mBMSCs using simple and convenient protocol, will aid preclinical studies based on the use of MSCs.
7.The repair effect and mechanism of amphiregulin on injured lung tissue in mice
Shuning TIAN ; Chen MENG ; Xianghong LUO ; Xianyu WANG
Chinese Journal of Emergency Medicine 2022;31(1):47-54
Objective:To investigate the repair effect of amphiregulin (Areg) on injured lung tissue in mice with acute respiratory distress syndrome (ARDS) and its underlying mechanism.Methods:The ARDS mouse model was made by tracheal infusion of lipopolysaccharide (LPS), and bronchoalveolar lavage fluid (BALF) was extracted for 7 consecutive days. Adult male C57BL/6 mice were randomly (random number) divided into 5 groups ( n=4 per group): (1) Control group; (2) Areg group: mice were treated intraperitoneally (i.p.) with recombinant Areg; (3) LPS+PBS group; (4) LPS+Areg group; and (5) LPS+Anti-Areg group; mice were instilled with LPS, then were injected i.p. with PBS, Areg or Areg neutralization antibody (Anti-Areg) 30 min later. Lung tissue and BALF were extracted at day 1, 3, 5 and 7 after ARDS. HE staining was used to evaluate the pathological changes of lung tissues. The total protein content in BALF was detected by BCA method, and the concentrations of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), IL-1β and immunoglobulin M (IgM) were determined by ELISA method. The phosphorylated levels of epidermal growth factor receptor (EGFR) and expressions of proliferating cell nuclear antigen (PCNA) and surface proteins-C (SP-C) were tested by Western blot. The immunofluorescence was used to detect the co-expression of PCNA and SP-C in lung tissues. One-way analysis of variance was used to compare the mean values of normally distributed measurement data between groups. Comparisons between groups were performed using the least significant difference t-test. Results:Compared with that at before modeling [(51.05±2.47) pg/mL], Areg concentrations were increased significantly at day 1 [(71.97±6.51) pg/mL; P<0.01] and day 3 [(147.58±7.56) pg/mL, P<0.01] in the BALF after ARDS. At day 1 after ARDS, there were significant interstitial edema, neutrophil infiltration and alveolar collapse in the LPS+PBS group and LPS+Areg group. Compared with the LPS+PBS group at day 3, 5 and 7, the pathological changes of lung tissues were notably improved in the LPS+Areg group, while were more serious in the LPS+Anti-Areg group. Compared with the control group, the LPS+PBS group had higher levels of neutrophil number, total protein, IgM, TNF-α, IL-1β, and IL-6. However, Areg treatment significantly reduced the levels of these indicators. Moreover, the expressions of PCNA (1.34±0.10), SP-C (1.48±0.10) and p-EGFR (0.92±0.032) in the LPS+Areg group were significantly up-regulated compared with those in the LPS+PBS group (0.88±0.03, 1.06±0.15, and 0.68±0.03, all P<0.01). And compared with the LPS+PBS group, PCNA and SP-C double positive cells were significantly increased in the LPS+Areg group, but decreased in the LPS+Anti-Areg group. Conclusions:Areg enhances the proliferation of alveolar typeⅡ epithelial cells by activating EGFR pathway, therefore promotes the repair of lung tissues during ARDS development.
8.Polypharmacy burden and the associated factors among inpatients with elderly comorbidity
Zhiyu WANG ; Xianyu LU ; Fengmei XING
Chinese Journal of Practical Nursing 2022;38(12):906-910
Objective:To explore the current status and influencing factors of polypharmacy burden in inpatient with elderly comorbidity.Methods:Convenience sampling method was used to select 393 in patients with elderly comorbidity of multiple medications who were hospitalized in North China University of Science and Technology Affiliated Hospital from September 2020 to January 2021 as the research object, using the general situation questionnaire, the Living with Medicines Questionnaire (LMQ), the Perceived Social Support Scale (PSSS), the Medication Compliance Scale (Morisky Scale) to investigate. Descriptive analysis of general data characteristics and polypharmacy burden scores were performed, and t-test, variance analysis, and multiple linear stepwise regression analysis were used to analyze influencing factors. Results:The polypharmacy burden for inpatient with elderly comorbidity was (136.95 ± 22.34) points, and the score rate was 70.23%, which was at a moderately high level. Multi-factor analysis showed that age, education level, personal monthly income, the number of illnesses, the number of medications, the duration of medication, social support, and medication compliance were independent factors influencing polypharmacy burden in inpatient with elderly comorbidity ( R 2=0.821, adjusted R 2=0.818, F=220.65, P<0.001). Conclusions:The polypharmacy burden in inpatient with elderly comorbidity is at a moderately high level. It is recommended that medical staff should develop individualized nursing interventions to reduce the polypharmacy burden for the factors that affect the burden of multiple medications for this population.
9.Role of epidermal growth factor in repair of lung tissues in mice with acute respiratory distress syndrome
Yongqian JIAO ; Chen MENG ; Wen ZENG ; Yiming WANG ; Silu WANG ; Xue WANG ; Nannan ZOU ; Xianyu WANG
Chinese Journal of Anesthesiology 2022;42(3):347-353
Objective:To evaluate the role of epidermal growth factor (EGF) in repair of lung tissues in mice with acute respiratory distress syndrome (ARDS).Methods:Fifty SPF male C57BL/6 mice, aged 6-8 weeks, weighing 21-23 g, were divided into 5 groups ( n=10 each) using a random number table method: control group (group C), EGF group, LPS+ PBS group, LPS+ EGF group and AG1478+ LPS+ EGF group.PBS 0.1 ml was intraperitoneally injected in group C. EGF 10 μg (0.1 ml) was intraperitoneally injected in group EGF.The equal volume of PBS and EGF 10 μg was intraperitoneally injected at 12 h after tracheal infusion of LPS in group LPS+ PBS and group LPS+ EGF, respectively.EGF receptor (EGFR) antagonist AG1478 1 mg was intraperitoneally injected, 30 min later LPS was tracheally instilled, and 12 h later EGF 10 μg was intraperitoneally injected in group AG1478+ LPS+ EGF.ARDS model was developed by endotracheal instillation of LPS 3 mg/kg.The mice were sacrificed on the 1st and 5th days after development of the model, and lung tissues were obtained for microscopic examination of the pathological changes which were scored after HE staining.Bronchoalveolar lavage was performed on 5th day after development of the model and before sacrifice, and bronchoalveolar lavage fluid (BALF) was collected to detect total protein concentration (by BCA method) and IL-6 and TNF-α concentrations (by enzyme-linked immunosorbent assay). Lung tissues were obtained for determination of the wet/dry lung weight ratio (W/D ratio), expression of lung surfactant associated protein C (SP-C) and proliferating nuclear antigen (PCNA) (by immunofluorescence method), and expression of EGFR, phosphorylated EGFR (p-EGFR), protein kinase B (Akt), and phosphorylated Akt (p-Akt) (by Western blot). Results:Compared with group C, the pathological score, W/D ratio, concentrations of total protein, IL-6 and TNF-α in BALF and neutrophil count were significantly increased, the number of cells co-expressing SP-C and PCNA was increased, and p-EGFR/EGFR and p-Akt/Akt ratios were increased in group LPS+ PBS ( P<0.01), and no significant change was found in the indexes mentioned above in group EGF ( P>0.05). Compared with group LPS+ PBS, the pathological score, W/D ratio, concentrations of total protein, IL-6 and TNF-α in BALF and neutrophil count were significantly decreased, the number of cells co-expressing SP-C and PCNA was increased, and p-EGFR/EGFR and p-Akt/Akt ratios were increased in group LPS+ EGF ( P<0.01). Compared with group LPS+ EGF, the pathological score, W/D ratio, concentrations of total protein, IL-6 and TNF-α in BALF and neutrophil count were significantly increased, the number of cells co-expressing SP-C and PCNA was decreased, and p-EGFR/EGFR and p-Akt/Akt ratios were decreased in group AG1478+ LPS+ EGF ( P<0.01). Conclusions:EGF can promote the repair of lung tissues in mice with ARDS, and the mechanism may be related to activation of EGFR signaling pathway and promotion of proliferation of alveolar epithelial cell type Ⅱ.
10.A novel test strip assay based on gold nanoparticles-labeled nucleic acid probe for rapid detection of Vibrio splendidus.
Lizhi ZHANG ; Ya'nan WEI ; Xuehui WANG ; Jingzhuang XUE ; Jiali QI ; Xianyu WANG ; Lei WANG ; Hongyan SU
Chinese Journal of Biotechnology 2021;37(9):3323-3333
Vibrio splendidus is an opportunistic pathogen in aquaculture. It can infect a variety of aquaculture animals and has caused huge losses to the aquaculture industry. In this study, a novel and efficient method for detecting V. splendidus was developed by combining the exonuclease Ⅲ amplification strategy with a nucleic acid test strip developed based on gold nanoparticles-labeled DNA probe. The results could be directly visualized by naked eyes, and this system overcame the difficulty in preparation of the monoclonal antibody used in conventional immunostrip. Upon optimization of experimental conditions, the detection limit of the strip was 5 ng/mL for the synthetic oligonucleotide DNA fragment and 10 ng/mL for the actual genomic DNA sample of V. splendidus. This test strip was more sensitive compared with the PCR method and was specific for the detection of V. splendidus. The rapid preparation of nucleic acid strip and the efficient detection of V. splendidus open a new way for the prevention and control of aquatic diseases.
Animals
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DNA Probes
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Gold
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Metal Nanoparticles
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Vibrio/genetics*

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