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1.
Hortic Res ; 10(7): uhad103, 2023 Jul.
Article En | MEDLINE | ID: mdl-37786729

Carrot (Daucus carota) is an Apiaceae plant with multi-colored fleshy roots that provides a model system for carotenoid research. In this study, we assembled a 430.40 Mb high-quality gapless genome to the telomere-to-telomere (T2T) level of "Kurodagosun" carrot. In total, 36 268 genes were identified and 34 961 of them were functionally annotated. The proportion of repeat sequences in the genome was 55.3%, mainly long terminal repeats. Depending on the coverage of the repeats, 14 telomeres and 9 centromeric regions on the chromosomes were predicted. A phylogenetic analysis showed that carrots evolved early in the family Apiaceae. Based on the T2T genome, we reconstructed the carotenoid metabolic pathway and identified the structural genes that regulate carotenoid biosynthesis. Among the 65 genes that were screened, 9 were newly identified. Additionally, some gene sequences overlapped with transposons, suggesting replication and functional differentiation of carotenoid-related genes during carrot evolution. Given that some gene copies were barely expressed during development, they might be functionally redundant. Comparison of 24 cytochrome P450 genes associated with carotenoid biosynthesis revealed the tandem or proximal duplication resulting in expansion of CYP gene family. These results provided molecular information for carrot carotenoid accumulation and contributed to a new genetic resource.

2.
Abdom Radiol (NY) ; 48(11): 3343-3352, 2023 11.
Article En | MEDLINE | ID: mdl-37495746

BACKGROUND: Hepatocellular carcinoma (HCC) is the sixth most common cancer, and the third leading cause of cancer death worldwide. Studies have shown that increased angiopoietin-2 (Ang-2) expression relative to Ang-1 expression in tumors is associated with a poor prognosis.The purpose of this study was to investigate the efficacy of predicting Ang-2 expression in HCC by preoperative dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI)-based radiomics. METHODS: The data of 52 patients with HCC who underwent surgical resection in our hospital were retrospectively analyzed. Ang-2 expression in HCC was analyzed by immunohistochemistry. All patients underwent preoperative upper abdominal DCE-MRI and intravoxel incoherent motion diffusion-weighted imaging scans. Radiomics features were extracted from the early and late arterial and portal phases of axial DCE-MRI. Univariate analysis and least absolute shrinkage and selection operator (LASSO) was performed to select the optimal radiomics features for analysis. A logistic regression analysis was performed to establish a DCE-MRI radiomics model, clinic-radiologic (CR) model and combined model integrating the radiomics score with CR factors. The stability of each model was verified by 10-fold cross-validation. Receiver operating characteristic (ROC) curve analysis, calibration curve analysis and decision curve analysis (DCA) were employed to evaluate these models. RESULTS: Among the 52 HCC patients, high Ang-2 expression was found in 30, and low Ang-2 expression was found in 22. The areas under the ROC curve (AUCs) for the radiomics model, CR model and combined model for predicting Ang-2 expression were 0.800, 0.874, and 0.933, respectively. The DeLong test showed that there was no significant difference in the AUC between the radiomics model and the CR model (p > 0.05) but that the AUC for the combined model was significantly greater than those for the other 2 models (p < 0.05). The DCA results showed that the combined model outperformed the other 2 models and had the highest net benefit. CONCLUSION: The DCE-MRI-based radiomics model has the potential to predict Ang-2 expression in HCC patients; the combined model integrating the radiomics score with CR factors can further improve the prediction performance.


Carcinoma, Hepatocellular , Liver Neoplasms , Humans , Carcinoma, Hepatocellular/diagnostic imaging , Carcinoma, Hepatocellular/surgery , Angiopoietin-2 , Retrospective Studies , Liver Neoplasms/diagnostic imaging , Liver Neoplasms/surgery , Magnetic Resonance Imaging
3.
BMC Plant Biol ; 23(1): 151, 2023 Mar 21.
Article En | MEDLINE | ID: mdl-36941578

BACKGROUND: Water shortage caused by global warming seriously affects the yield and quality of vegetable crops. ß-carotene, the lipid-soluble natural product with important pharmacological value, is abundant in celery. Transcription factor MYB family extensively disperses in plants and plays regulatory roles in carotenoid metabolism and water scarcity response. RESULTS: Here, the AgMYB5 gene encoding 196 amino acids was amplified from celery cv. 'Jinnanshiqin'. In celery, the expression of AgMYB5 exhibited transactivation activity, tissue specificity, and drought-condition responsiveness. Further analysis proved that ectopic expression of AgMYB5 increased ß-carotene content and promoted drought tolerance in transgenic Arabidopsis thaliana. Moreover, AgMYB5 expression promoted ß-carotene biosynthesis by triggering the expression of AtCRTISO and AtLCYB, which in turn increased antioxidant enzyme activities, and led to the decreased contents of H2O2 and MDA, and the inhibition of O2- generation. Meanwhile, ß-carotene accumulation promoted endogenous ABA biosynthesis of transgenic Arabidopsis, which resulted in ABA-induced stomatal closing and delayed water loss. In addition, ectopic expression of AgMYB5 increased expression levels of AtERD1, AtP5CS1, AtRD22, and AtRD29. CONCLUSIONS: The findings indicated that AgMYB5 up-regulated ß-carotene biosynthesis and drought tolerance of Arabidopsis.


Apium , Arabidopsis , Arabidopsis/metabolism , beta Carotene , Apium/genetics , Apium/metabolism , Drought Resistance , Transcription Factors/genetics , Transcription Factors/metabolism , Vegetables/genetics , Vegetables/metabolism , Hydrogen Peroxide/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Plants, Genetically Modified/genetics , Plants, Genetically Modified/metabolism , Stress, Physiological/genetics , Antioxidants/metabolism , Droughts , Water/metabolism , Gene Expression Regulation, Plant , Abscisic Acid/metabolism
4.
Journal of Forensic Medicine ; (6): 137-143, 2023.
Article En | WPRIM | ID: wpr-981847

OBJECTIVES@#To explore the changes of elbow flexor muscle strength after musculocutaneous nerve injury and its correlation with needle electromyography (nEMG) parameters.@*METHODS@#Thirty cases of elbow flexor weakness caused by unilateral brachial plexus injury (involving musculocutaneous nerve) were collected. The elbow flexor muscle strength was evaluated by manual muscle test (MMT) based on Lovett Scale. All subjects were divided into Group A (grade 1 and grade 2, 16 cases) and Group B (grade 3 and grade 4, 14 cases) according to their elbow flexor muscle strength of injured side. The biceps brachii of the injured side and the healthy side were examined by nEMG. The latency and amplitude of the compound muscle action potential (CMAP) were recorded. The type of recruitment response, the mean number of turns and the mean amplitude of recruitment potential were recorded when the subjects performed maximal voluntary contraction. The quantitative elbow flexor muscle strength was measured by portable microFET 2 Manual Muscle Tester. The percentage of residual elbow flexor muscle strength (the ratio of quantitative muscle strength of the injured side to the healthy side) was calculated. The differences of nEMG parameters, quantitative muscle strength and residual elbow flexor muscle strength between the two groups and between the injured side and the healthy side were compared. The correlation between elbow flexor manual muscle strength classification, quantitative muscle strength and nEMG parameters was analyzed.@*RESULTS@#After musculocutaneous nerve injury, the percentage of residual elbow flexor muscle strength in Group B was 23.43% and that in Group A was 4.13%. Elbow flexor manual muscle strength classification was significantly correlated with the type of recruitment response, and the correlation coefficient was 0.886 (P<0.05). The quantitative elbow flexor muscle strength was correlated with the latency and amplitude of CMAP, the mean number of turns and the mean amplitude of recruitment potential, and the correlation coefficients were -0.528, 0.588, 0.465 and 0.426 (P<0.05), respectively.@*CONCLUSIONS@#The percentage of residual elbow flexor muscle strength can be used as the basis of muscle strength classification, and the comprehensive application of nEMG parameters can be used to infer quantitative elbow flexor muscle strength.


Humans , Elbow , Electromyography , Musculocutaneous Nerve , Elbow Joint/physiology , Muscle, Skeletal , Muscle Strength , Peripheral Nerve Injuries
5.
Signal Transduct Target Ther ; 7(1): 382, 2022 11 25.
Article En | MEDLINE | ID: mdl-36424379

COVID-19 patients can develop clinical and histopathological features associated with fibrosis, but the pathogenesis of fibrosis remains poorly understood. CD147 has been identified as a universal receptor for SARS-CoV-2 and its variants, which could initiate COVID-19-related cytokine storm. Here, we systemically analyzed lung pathogenesis in SARS-CoV-2- and its delta variant-infected humanized CD147 transgenic mice. Histopathology and Transmission Electron Microscopy revealed inflammation, fibroblast expansion and pronounced fibrotic remodeling in SARS-CoV-2-infected lungs. Consistently, RNA-sequencing identified a set of fibrosis signature genes. Furthermore, we identified CD147 as a crucial regulator for fibroblast activation induced by SARS-CoV-2. We found conditional knockout of CD147 in fibroblast suppressed activation of fibroblasts, decreasing susceptibility to bleomycin-induced pulmonary fibrosis. Meplazumab, a CD147 antibody, was able to inhibit the accumulation of activated fibroblasts and the production of ECM proteins, thus alleviating the progression of pulmonary fibrosis caused by SARS-CoV-2. In conclusion, we demonstrated that CD147 contributed to SARS-CoV-2-triggered progressive pulmonary fibrosis and identified CD147 as a potential therapeutic target for treating patients with post-COVID-19 pulmonary fibrosis.


COVID-19 , Pulmonary Fibrosis , Mice , Animals , Pulmonary Fibrosis/genetics , SARS-CoV-2 , COVID-19/genetics
6.
Redox Biol ; 57: 102509, 2022 Nov.
Article En | MEDLINE | ID: mdl-36302319

Idiopathic pulmonary fibrosis (IPF) is a chronic progressive disease characterized by excessive proliferation of fibroblasts and excessive accumulation of extracellular matrix (ECM). Ferroptosis is a novel form of cell death characterized by the lethal accumulation of iron and lipid peroxidation, which is associated with many diseases. Our study addressed the potential role played by ferroptosis and iron accumulation in the progression of pulmonary fibrosis. We found that the inducers of pulmonary fibrosis and injury, namely, bleomycin (BLM) and lipopolysaccharide (LPS), induced ferroptosis of lung epithelial cells. Both the ferroptosis inhibitor liproxstatin-1 (Lip-1) and the iron chelator deferoxamine (DFO) alleviated the symptoms of pulmonary fibrosis induced by bleomycin or LPS. TGF-ß stimulation upregulated the expression of transferrin receptor protein 1 (TFRC) in the human lung fibroblast cell line (MRC-5) and mouse primary lung fibroblasts, resulting in increased intracellular Fe2+, which promoted the transformation of fibroblasts into myofibroblasts. Mechanistically, TGF-ß enhanced the expression and nuclear localization of the transcriptional coactivator tafazzin (TAZ), which combined with the transcription factor TEA domain protein (TEAD)-4 to promote the transcription of TFRC. In addition, elevated Fe2+ failed to induce the ferroptosis of fibroblasts, which might be related to the regulation of iron export and lipid metabolism. Finally, we specifically knocked out TFRC expression in fibroblasts in mice, and compared with those in the control mice, the symptoms of pulmonary fibrosis were reduced in the knockout mice after bleomycin induction. Collectively, these findings suggest the therapeutic potential of ferroptosis inhibitors and iron chelators in treating pulmonary fibrosis.

7.
J Food Biochem ; 46(12): e14403, 2022 12.
Article En | MEDLINE | ID: mdl-36121702

Walnut diaphragm is defined as a dry wood septum located between the walnut shell and kernel. In this work, seven phenolic compounds from walnut diaphragm were purified and characterized, and their antioxidant activities and mechanisms of hypoglycemia were investigated. Compounds 1-7 were tested for DPPH, ABTS scavenging ability, and FRAP assay to evaluate the antioxidant activity. α-Amylase inhibition assay was introduced to assess the hypoglycemic activity, and the mechanism was investigated by kinetic analysis, CD spectrum, and molecular docking. Compound 6 showed the strongest antioxidant ability, while compound 1 exhibited the strongest inhibition of α-amylase by changing the secondary structure of α-amylase in a mixed competitive inhibition mode. Molecular docking test predicted that the tetrahydropyran part in compound 1 may contribute to its hypoglycemic effect. This study furnishes a new theoretical reference for the utilization and development of walnut diaphragm into a health food with antioxidant and hypoglycemic properties. PRACTICAL APPLICATIONS: The finding of this research may serve as a basis for the subsequent development of walnut diaphragm into instant tea-based health food or added to other food carriers to achieve auxiliary antioxidant and hypoglycemic effects. This study revealed that polyphenolic components were the material basis for the antioxidant and hypoglycemic effects of walnut diaphragm, which could be identified as landmark chemical components for controlling quality standards in the development of walnut diaphragm, thus accelerating the research process of quality standards for walnut diaphragm-related products. Furthermore, the studies on the mechanism of hypoglycemic activity supply more credible data to support the development of walnut diaphragm into a safe and consumer-friendly health food. With abundant resources and clear efficacy, walnut diaphragm has great development prospect and application value.


Antioxidants , Juglans , Antioxidants/chemistry , Juglans/chemistry , Hypoglycemic Agents/pharmacology , Hypoglycemic Agents/chemistry , Diaphragm/chemistry , Diaphragm/metabolism , Molecular Docking Simulation , Kinetics , Plant Extracts/pharmacology , Plant Extracts/chemistry , Phenols/analysis , alpha-Amylases/metabolism
8.
Nat Commun ; 13(1): 676, 2022 02 03.
Article En | MEDLINE | ID: mdl-35115492

Ferroptosis is a nonapoptotic cell death process that requires cellular iron and the accumulation of lipid peroxides. In progressive rheumatoid arthritis (RA), synovial fibroblasts proliferate abnormally in the presence of reactive oxygen species (ROS) and elevated lipid oxidation. Here we show, using a collagen-induced arthritis (CIA) mouse model, that imidazole ketone erastin (IKE), a ferroptosis inducer, decreases fibroblast numbers in the synovium. Data from single-cell RNA sequencing further identify two groups of fibroblasts that have distinct susceptibility to IKE-induced ferroptosis, with the ferroptosis-resistant fibroblasts associated with an increased TNF-related transcriptome. Mechanistically, TNF signaling promotes cystine uptake and biosynthesis of glutathione (GSH) to protect fibroblasts from ferroptosis. Lastly, low dose IKE together with etanercept, a TNF antagonist, induce ferroptosis in fibroblasts and attenuate arthritis progression in the CIA model. Our results thus imply that the combination of TNF inhibitors and ferroptosis inducers may serve as a potential candidate for RA therapy.


Arthritis, Experimental/prevention & control , Arthritis, Rheumatoid/prevention & control , Ferroptosis/drug effects , Fibroblasts/drug effects , Imidazoles/pharmacology , Ketones/pharmacology , Piperazines/pharmacology , Tumor Necrosis Factor Inhibitors/pharmacology , Animals , Arthritis, Experimental/genetics , Arthritis, Experimental/metabolism , Arthritis, Rheumatoid/genetics , Arthritis, Rheumatoid/metabolism , Cells, Cultured , Disease Models, Animal , Drug Therapy, Combination , Etanercept/pharmacology , Etanercept/therapeutic use , Fibroblasts/cytology , Fibroblasts/metabolism , Glutathione/metabolism , Humans , Imidazoles/therapeutic use , Ketones/therapeutic use , Lipid Peroxidation/drug effects , Mice , Piperazines/therapeutic use , Reactive Oxygen Species/metabolism , Synovial Membrane/cytology , Tumor Necrosis Factor Inhibitors/therapeutic use
9.
Article Zh | WPRIM | ID: wpr-936262

Objective: To analyze the incidence and risk factors of otologic disorders in patients with Turner syndrome (TS), so as to provide management strategies for ear health. Methods: This study is a prospective study based on questionnaires and a cross-sectional study. The TS patients who visited our hospital from 2010 January to 2021 March were included (A total of 71 patients with TS were included in this study. the age of TS diagnosed was 3- to 11-year-old, age of visiting ENT department was 4- to 27-year-old) and the incidence of otologic diseases in different age groups was investigated by questionnaires. The cross-sectional study included ear morphology and auditory function assessment, and further analysis of the risk factors that related to ear disease. Prism was used for data analysis. Results: The investigation found that the incidence of acute otitis media in patients aged 3-6 and 7-12 years was higher than that of patients over 12 years old, which was 33.8%(24/71), 42.9%(30/70)and 23.5%(8/34), respectively; 21.1% (15/71) of patients were recurrent acute otitis media in patients aged 3-6 years, and about 46.6% (7/15)of them persisted beyond 6-year. The prevalence of otitis media with effusion in the three groups was 32.4%(23/71), 34.3%(24/70)and 38.2%(13/34), respectively; the recurrence rate of tympanocentesis was 100%(7/7), 42.9%(3/7)and 50.0%(1/2), which was significantly higher than that of grommet insertion. For age groups of 3-6 and 7-12 years, the prevalence of acute otitis media and secretory otitis media was lower in the X chromosome structure abnormal patients; while for patients older than 12 years, otitis media with effusion was the highest prevalence in Y-chromosome-containing karyotypes. In addition, the prevalence of acute otitis media and otitis media with effusion in patients with other system diseases were increased significantly. A cross-sectional study found that 7.0% (5/71)of the lower auricular, 4.2% (3/71)of the external auditory canal narrow, and 38.0% (27/71)of the tympanic membrane abnormality. 35.2%(25/71) had abnormal hearing, including 17 cases of conductive deafness, 6 cases of sensorineural hearing loss, and 2 cases of mixed deafness. The rest of the patients had normal hearing, but 6 of them had abnormalities in otoacoustic emission. Eustachian tube function assessment found that the eustachian tube dysfunction accounted for 38%(27/71). Hearing loss and abnormal Eustachian tube function were not significantly related to karyotype(Chi-square 2.83 and 2.84,P value 0.418 and 0.417), but significantly related to other system diseases(Chi-square 13.43 and 7.53,P value<0.001). Conclusions: The incidence of TS-related otitis media and auditory dysfunction is significantly higher than that of the general population. It not only occurs in preschool girls, but also persists or develops after school age. Accompanied by other system diseases are risk factors for ear diseases. Clinicians should raise their awareness of TS-related ear diseases and incorporate ear health monitoring into routine diagnosis and treatment.


Adolescent , Adult , Child , Child, Preschool , Female , Humans , Young Adult , Cross-Sectional Studies , Deafness/etiology , Middle Ear Ventilation/adverse effects , Otitis Media/complications , Otitis Media with Effusion/complications , Prospective Studies , Turner Syndrome/therapy
10.
Signal Transduct Target Ther ; 6(1): 347, 2021 09 25.
Article En | MEDLINE | ID: mdl-34564690

SARS-CoV-2 mutations contribute to increased viral transmissibility and immune escape, compromising the effectiveness of existing vaccines and neutralizing antibodies. An in-depth investigation on COVID-19 pathogenesis is urgently needed to develop a strategy against SARS-CoV-2 variants. Here, we identified CD147 as a universal receptor for SARS-CoV-2 and its variants. Meanwhile, Meplazeumab, a humanized anti-CD147 antibody, could block cellular entry of SARS-CoV-2 and its variants-alpha, beta, gamma, and delta, with inhibition rates of 68.7, 75.7, 52.1, 52.1, and 62.3% at 60 µg/ml, respectively. Furthermore, humanized CD147 transgenic mice were susceptible to SARS-CoV-2 and its two variants, alpha and beta. When infected, these mice developed exudative alveolar pneumonia, featured by immune responses involving alveoli-infiltrated macrophages, neutrophils, and lymphocytes and activation of IL-17 signaling pathway. Mechanistically, we proposed that severe COVID-19-related cytokine storm is induced by a "spike protein-CD147-CyPA signaling axis": Infection of SARS-CoV-2 through CD147 initiated the JAK-STAT pathway, which further induced expression of cyclophilin A (CyPA); CyPA reciprocally bound to CD147 and triggered MAPK pathway. Consequently, the MAPK pathway regulated the expression of cytokines and chemokines, which promoted the development of cytokine storm. Importantly, Meplazumab could effectively inhibit viral entry and inflammation caused by SARS-CoV-2 and its variants. Therefore, our findings provided a new perspective for severe COVID-19-related pathogenesis. Furthermore, the validated universal receptor for SARS-CoV-2 and its variants can be targeted for COVID-19 treatment.


Angiotensin-Converting Enzyme 2/metabolism , Antibodies, Monoclonal, Humanized/pharmacology , Basigin/antagonists & inhibitors , Basigin/metabolism , COVID-19 Drug Treatment , COVID-19/metabolism , Cytokine Release Syndrome/drug therapy , SARS-CoV-2/metabolism , Angiotensin-Converting Enzyme 2/genetics , Animals , Basigin/genetics , COVID-19/genetics , Chlorocebus aethiops , Cytokine Release Syndrome/genetics , Cytokine Release Syndrome/metabolism , Humans , MAP Kinase Signaling System/drug effects , MAP Kinase Signaling System/genetics , Mice , Mice, Transgenic , SARS-CoV-2/genetics , Vero Cells
11.
Genes Dis ; 8(3): 364-372, 2021 May.
Article En | MEDLINE | ID: mdl-33997183

Circular RNAs are a large class of noncoding RNAs. Smad5 functions in cell differentiation, cell proliferation and metastasis. It has been reported that lnc-Smad5 can inhibit the proliferation of diffuse large B cell lymphoma. However, the function of circ-Smad5 has not yet been reported. Lentivirus vectors were constructed to establish circ-Smad5 upregulated and circ-Smad5 downregulated cell models. A CCK-8 assay was used to detect the proliferation of JB6 cells. FACS was used to analyze the cell cycle in the cell models. Western blot, immunofluorescence staining and TOP/FOP flash dual luciferase activity assays were used to determine the activity of the Wnt signaling pathway. The results revealed that the expression level of circ-Smad5 in JB6 cells was significantly lower than the expression level of linearized-Smad5. Compared with the control group, the percentage of S phase cells and the expression level of cyclin D1 protein were significantly higher in the sh-circ-Smad5 group. In the sh-circ-Smad5 group, ß-catenin and LEF-1 were significantly increased, p-ß-catenin was significantly decreased, and the relative activity of the TOP/FOP reporter gene was higher compared to the control group levels. These phenomena could be reversed by treating with Wnt signaling inhibitor PNU-74654. We conclude that the circ-Smad5 retards the proliferation and the cell cycle progression of JB6 cells. Thus, circ-Smad5 may function by inhibiting the activation of Wnt/ß-catenin/Lef 1 signaling, which inhibits the expression of cyclin D1. To the best of our knowledge, we are the first to report the function of circ-Smad5.

12.
Front Cell Dev Biol ; 9: 751593, 2021.
Article En | MEDLINE | ID: mdl-34977009

Ferroptosis, a form of programmed cell death process driven by iron-dependent lipid peroxidation, plays an important role in tumor suppression. Although previous study showed that intracellular Merlin-Hippo signaling suppresses ferroptosis of epithelial tumor cells through the inactivation of YAP signaling, it remains elusive if the proto-oncogenic transcriptional co-activator YAP could serve as a potential biomarker to predict cancer cell response to ferroptosis-inducing therapies. In this study, we show that both total YAP staining and nuclear YAP staining were more prevalent in HCC tissues than in nontumorous regions. Compared to low-density HCC cells, high-density cells showed decreased nuclear localization of YAP and conferred significant resistance to ferroptosis. Oncogenic activation of YAP signaling by overexpression of YAP(S127A) mutant sensitized ferroptosis of HCC cells cultured in confluent density or in the 3D tumor spheroid model. Furthermore, we validated the lipoxygenase ALOXE3 as a YAP-TEAD target gene that contributed to YAP-promoted ferroptosis. Overexpression of ALOXE3 effectively increased the vulnerability of HCC cells to ferroptotic cell death. In an orthotopic mouse model of HCC, genetic activation of YAP rendered HCC cells more susceptible to ferroptosis. Finally, an overall survival assay further revealed that both a high expression of YAP and a low expression of GPX4 were correlated with increased survival of HCC patients with sorafenib treatment, which had been proven to be an inducer for ferroptosis by inhibition of the xc-amino acid antiporter. Together, this study unveils the critical role of intracellular YAP signaling in dictating ferroptotic cell death; it also suggests that pathogenic alterations of YAP signaling can serve as biomarkers to predict cancer cell responsiveness to future ferroptosis-inducing therapies.

14.
Anal Biochem ; 599: 113735, 2020 06 15.
Article En | MEDLINE | ID: mdl-32294460

In view of the current demand for rapid detection and identification of pathogens, point-of-care testing (POCT) with fast portability, low consumption, and increased sensitivity and specificity has become more and more popular. The emerging nucleic acid isothermal amplification technology (NAIAT) has shown potential advantages in the development of rapid microbial detection. In this study, a micro-detection slide system was developed based on the NAIAT of various nucleic acids of shrimp pathogens. The system included a micro-detection slide with 48 identical detecting cells precoated with all detection reagents, except the sample template. The process of producing the micro-detection slides mainly combined super-hydrophobic/super-oleophobic and super-hydrophilic materials to obtain separated spaces for detection, and aerosol pollution was eliminated in the form of water-in-oil. The micro-detection slide system was capable of simultaneously detecting 4 groups of samples and 8 important shrimp pathogens and is a relatively low-cost, portable, and high-throughput nucleic acid (RNA and DNA) detection technology. The establishment of this technology will provide key technical support for the construction of biosecurity systems for healthy shrimp culture.


Animal Diseases , Nucleic Acid Amplification Techniques/methods , Penaeidae , Animal Diseases/diagnosis , Animal Diseases/microbiology , Animal Diseases/parasitology , Animal Diseases/virology , Animals , Nucleic Acid Amplification Techniques/veterinary , Nucleic Acids/analysis , Penaeidae/microbiology , Penaeidae/parasitology , Penaeidae/virology
15.
Hum Gene Ther Methods ; 30(1): 1-16, 2019 02.
Article En | MEDLINE | ID: mdl-30700146

As a nucleic acid alternative to traditional antibody, aptamer holds great potential in various fields of biology and medicine such as targeted gene therapy, drug delivery, bio-sensing, and laboratory medicine. Over the past decades, the conventional Systematic Evolution of Ligands by Exponential Enrichment (SELEX) method has undergone dramatic modifications and improvements owing to developments in material sciences and analytical techniques. However, many of the recently developed strategies either require complex materials and instruments or suffer from low efficiency and high failure rates in the selection of desired aptamers. Accordingly, the development of aptamers against new or novel targets is still a major obstacle for aptamer-based research and application. Here, an improved protein-SELEX procedure is presented for simplified and highly efficient isolation of aptamers against protein targets. Approaches are described that ensure a high success rate in aptamer selection by simplifying polymerase chain reaction procedures, introducing denature gel, utilizing an electro-elution-based single-stranded DNA separation strategy, as well as an enzyme-linked immunosorbent assay-based highly sensitive binding assay. In addition, a simplified sample preparation method for MiSeq-based next-generation sequencing is also introduced. While a recombinant protein as a bait protein for SELEX is discussed here, this protocol will also be invaluable for researchers wishing to develop aptamers against targets other than proteins such as small molecules, lipids, carbohydrates, cells, and micro-organisms for future gene therapy and/or diagnostics.


Aptamers, Nucleotide/chemistry , SELEX Aptamer Technique/methods , DNA, Single-Stranded/isolation & purification , Enzyme-Linked Immunosorbent Assay/methods , Gene Library , Genetic Therapy/methods , High-Throughput Nucleotide Sequencing/methods , Humans , Polymerase Chain Reaction , Recombinant Proteins
16.
Journal of Forensic Medicine ; (6): 359-362, 2018.
Article En | WPRIM | ID: wpr-984943

OBJECTIVES@#To explore the assessment method of original height of L1-2 after vertebral compression fracture and its application value in forensic clinical practice.@*METHODS@#A total of 154 normal thoracic and lumbar X-ray films were collected, and 140 cases were used as experimental group while 14 cases as validation group. The heights of anterior (Ha) and posterior (Hp) vertebral body of T₁₂-L₃ vertebrae in each X-ray image were measured. In the experimental group, the correlation analysis between HaL₁ and HaT₁₂, HpT₁₂, HpL₁, HaL₂ and HpL₂ was carried out, and regression equation was established via fitting. The correlation analysis between HaL₂ and HaL₁, HpL₁, HpL₂, HaL₃, HpL₃ was performed, and the regression equation was also established via fitting. The difference between the predicted and measured values of HaL₁ and HaL₂ in validation group was compared.@*RESULTS@#In the 140 normal subjects, HaL₁ (y₁) was well correlated with HaT₁₂ (x₁) and HaL₂(x₂), and the multiple linear regression equation was y₁=2.545+0.423 x₁+0.486 x₂ (determining coefficient R²=0.712, P<0.05; F=169.206, P<0.05). There was no significant difference between the predicted and actual measured values of HaL₁ in the validation group ( P>0.05). HaL₂ (y₂) was well correlated with HaL₁ (x₃) and HaL₃ (x₄), and the multiple linear regression equation was y₂=4.354+0.530 x₃+0.349 x₄ (determining coefficient R²=0.689, P<0.05; F=151.575, P<0.05). There was no significant difference between the predicted and actual measured values of HaL₂ in the validation group ( P>0.05).@*CONCLUSIONS@#It is more appropriate to evaluate the original height of L₁ or L₂ single vertebrae by comparing with the height of the anterior edge of the upper and lower adjacent vertebral bodies.


Aged , Humans , Middle Aged , Fractures, Compression , Lumbar Vertebrae/diagnostic imaging , Spinal Fractures/surgery , Thoracic Vertebrae/diagnostic imaging
17.
Journal of Forensic Medicine ; (6): 665-671, 2018.
Article Zh | WPRIM | ID: wpr-742816

Accurate evaluation of muscle function helps to understand the recovery of muscle, bone, nervous system diseases or injuries, especially for muscle dysfunction caused by peripheral nerve injury.Therefore, the methods of muscle function evaluation have been the focus of researchers, with new methods having been constantly proposed.Muscle strength testing is an important part of muscle function evaluation.Besides hand muscle strength assessment, currently used muscle function assessments include simple instrumental test, isokinetic muscle test, electrophysiological test, etc.In addition, the application of needle electromyography, motor unit number estimation, motion unit index in muscle function evaluation has also been reported for several times.This paper reviews the research progress and practical application of these methods.

18.
Theranostics ; 7(17): 4071-4086, 2017.
Article En | MEDLINE | ID: mdl-29158811

Chemotherapy-resistant cancer stem cells (CSCs) are a major obstacle to the effective treatment of many forms of cancer. To overcome CSC chemo-resistance, we developed a novel system by conjugating a CSC-targeting EpCAM aptamer with doxorubicin (Apt-DOX) to eliminate CSCs. Incubation of Apt-DOX with colorectal cancer cells resulted in high concentration and prolonged retention of DOX in the nuclei. Treatment of tumour-bearing xenograft mice with Apt-DOX resulted in at least 3-fold more inhibition of tumour growth and longer survival as well as a 30-fold lower frequency of CSC and a prolonged longer tumourigenic latency compared with those receiving the same dose of free DOX. Our data demonstrate that a CSC-targeting aptamer is able to transform a conventional chemotherapeutic agent into a CSC-killer to overcome drug resistance in solid tumours.


Aptamers, Nucleotide/administration & dosage , Doxorubicin/administration & dosage , Drug Delivery Systems/methods , Epithelial Cell Adhesion Molecule/genetics , Neoplastic Stem Cells/drug effects , Animals , Antibiotics, Antineoplastic/administration & dosage , Antibiotics, Antineoplastic/chemistry , Dose-Response Relationship, Drug , Doxorubicin/chemistry , Drug Carriers/administration & dosage , Drug Carriers/chemistry , Female , HT29 Cells , Humans , Hydrogen-Ion Concentration , Male , Mice, SCID , Polyethylene Glycols/chemistry , Rats, Sprague-Dawley , Xenograft Model Antitumor Assays
19.
Sci Rep ; 7(1): 5898, 2017 07 19.
Article En | MEDLINE | ID: mdl-28724889

The development of chemoresistance and inability in elimination of cancer stem cells are among the key limitations of cancer chemotherapy. Novel molecular therapeutic strategies able to overcome such limitations are urgently needed for future effective management of cancer. In this report, we show that EpCAM-aptamer-guided survivin RNAi effectively downregulated survivin both in colorectal cancer cells in vitro and in a mouse xenograft model for colorectal cancer. When combined with the conventional chemotherapeutic agents, the aptamer-guided survivin RNAi was able to enhance the sensitivity towards 5-FU or oxaliplatin in colorectal cancer stem cells, increase apoptosis, inhibit tumour growth and improve the overall survival of mice bearing xenograft colorectal cancer. Our results indicate that survivin is one of the key players responsible for the innate chemoresistance of colorectal cancer stem cells. Thus, aptamer-mediated targeting of survivin in cancer stem cells in combination with chemotherapeutic drugs constitutes a new avenue to improve treatment outcome in oncologic clinics.


Aptamers, Nucleotide/metabolism , Colorectal Neoplasms/pathology , Fluorouracil/pharmacology , Neoplastic Stem Cells/pathology , RNA Interference , Survivin/metabolism , Animals , Apoptosis/drug effects , Base Sequence , Cell Line, Tumor , Colorectal Neoplasms/metabolism , Down-Regulation/drug effects , Epithelial Cell Adhesion Molecule/metabolism , Female , Gene Knockdown Techniques , Humans , Mice, SCID , Neoplastic Stem Cells/drug effects , Neoplastic Stem Cells/metabolism
20.
Journal of Forensic Medicine ; (6): 353-356, 2017.
Article Zh | WPRIM | ID: wpr-667366

Objective To compare the differences between Hertel exophthalmometry and CT on exophthalmos measurement,and explore its practical application value in forensic medicine.Methods Fifty-six normal individuals without eye injuries or diseases were selected as the control group.The absolute values of exophthalmos were measured in the standardized CT image workstation.The difference of binocular exophthalmos was compared in normal group.Forty-seven cases with unilateral orbital fracture were divided into injured eye group and normal eye group according to the presence of binocular orbital fractures.The differences of the absolute values of eophthalmos from the same eye and the relative values of eophthalmos between two eyes for same person measured by Hertel exophthalmometry and CT were analyzed.Results There was no statistical difference of exophthalmos between the normal eyes measured by CT method.In the normal eye group,the absolute value of exophthalmos measured by CT method was (16.66±5.41) rm,which was (16.16±4.45)mm when measured by the Hertel exophthalmometry method.There was no statistical difference between two groups (P>0.05).In the injured eye group,there was statistical difference between the absolute values of exophthalmos measured by two measurement methods (P<0.05).There was no statistical difference of the relative values of exophthalmos between the injured eyes and the normal eyes by two measurement methods (P>0.05).Conclusion CT method has a good consistency with Hertel exophthalmometry,which can be applied into the practice of medicolegal expertise.

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