1.Cloning and Functional Characterization of Farnesyl Diphosphate Synthase Gene in Biosynthesis of Terpenoid Components in Chinese Materia Medica
Yue ZHANG ; Feng ZHANG ; Yue ZHANG ; Chaoyue LIU ; Bolin ZHANG ; Jia LIU ; Caixia WANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):175-183
ObjectiveThis study aims to enhance of the farnesyl pyrophosphate(FPP) pool in Saccharomyces cerevisiae by heterologously expressing different farnesyl diphosphate synthases(FPSs) from various plants, thereby increasing the production of terpenoid compounds by the engineered yeast. MethodsRNA from mixed samples of roots, stems, and leaves of seven plants including Arabidopsis thaliana, Rosa rugosa, Artemisia annua, Centella asiatica, Humulus lupulus, Medicago sativa, and Panax ginseng was extracted by column chromatography and reverse transcribed into the first strand of complementary DNA(cDNA), and based on the transcriptome data of the seven species of plants, sequence-specific primers were designed for CaFPS, RrFPS, MsFPS, HiFPS, PgFPS, AtFPS, and AaFPS, the full-length of the genes was cloned, and the genes were analyzed for bioinformatics in order to construct a pESC yeast shuttle vector. These seven plant-derived FPSs were further heterologously expressed in the previous constructed β-elemene-producing yeast, and the yield of β-elemene was indicated for their catalytic acivities. ResultsThe coding sequences of CaFPS, RrFPS, MsFPS, HiFPS, PgFPS, AtFPS, and AaFPS were all of 1 021 bp in length and encoding 301 amino acids, all of which were similarly related to the endogenous FPS-encoding gene(ERG20) in S. cerevisiae. After heterologous expression, RrFPS was identified as the most effective in catalyzing the synthesis of FPP from isopentenyl pyrophosphate(IPP) and dimethylallyl pyrophosphate(DMAPP). Compared to the control strains, the RrFPS overexpressed yeast strains YB-1-Rr and YB-3-Rr increased the production of β-elemene by 231.25% and 189.3%, respectively. ConclusionBy comparing the functions of FPS-encoding genes from seven different plant sources, it is determined that the protein encoded by the RrFPS from R. rugosa has the best catalytic ability, which can provide key genetic elements for the construction of engineered yeast strain constructs with high terpenoid production.
2.The role of intraplaque neovascularization in recent and future ischemic stroke in patients with mild carotid stenosis
Luni ZHANG ; Rong WU ; Jing CHEN ; Shiyao GU ; Caixia JIA
Ultrasonography 2025;44(1):62-71
Purpose:
There is still insufficient evidence for predicting stroke risk in patients with mild carotid atherosclerotic stenosis. This study aimed to explore the association between carotid intraplaque neovascularization (IPN) in mild stenosis and ischemic stroke, using contrast-enhanced ultrasound (CEUS) imaging.
Methods:
This retrospective observational study included 369 patients from July 2021 to March 2022. These patients were categorized as symptomatic or asymptomatic based on their recent history of ipsilateral ischemic stroke. Initial parameters of carotid plaques, such as IPN grading and contrast enhancement index, were assessed using B-mode ultrasonography and CEUS. The follow-up period lasted 12 months or until a newly-developed ischemic stroke occurred. Logistic regression models and Cox proportional-hazards regression models were employed to explore the associations between ultrasonic parameters and the incidence of recent and future ischemic strokes.
Results:
In patients with mild stenosis, both increasing age and grade 2 carotid IPN were significant predictors of recent primary ischemic stroke. Furthermore, grade 2 carotid IPN independently predicted future ischemic strokes in both symptomatic and asymptomatic patients.
Conclusion
This study demonstrated that carotid IPN as detected by CEUS imaging holds potential as a useful non-invasive biomarker for predicting recent and future ischemic strokes in patients with mild carotid stenosis.
3.The role of intraplaque neovascularization in recent and future ischemic stroke in patients with mild carotid stenosis
Luni ZHANG ; Rong WU ; Jing CHEN ; Shiyao GU ; Caixia JIA
Ultrasonography 2025;44(1):62-71
Purpose:
There is still insufficient evidence for predicting stroke risk in patients with mild carotid atherosclerotic stenosis. This study aimed to explore the association between carotid intraplaque neovascularization (IPN) in mild stenosis and ischemic stroke, using contrast-enhanced ultrasound (CEUS) imaging.
Methods:
This retrospective observational study included 369 patients from July 2021 to March 2022. These patients were categorized as symptomatic or asymptomatic based on their recent history of ipsilateral ischemic stroke. Initial parameters of carotid plaques, such as IPN grading and contrast enhancement index, were assessed using B-mode ultrasonography and CEUS. The follow-up period lasted 12 months or until a newly-developed ischemic stroke occurred. Logistic regression models and Cox proportional-hazards regression models were employed to explore the associations between ultrasonic parameters and the incidence of recent and future ischemic strokes.
Results:
In patients with mild stenosis, both increasing age and grade 2 carotid IPN were significant predictors of recent primary ischemic stroke. Furthermore, grade 2 carotid IPN independently predicted future ischemic strokes in both symptomatic and asymptomatic patients.
Conclusion
This study demonstrated that carotid IPN as detected by CEUS imaging holds potential as a useful non-invasive biomarker for predicting recent and future ischemic strokes in patients with mild carotid stenosis.
4.The role of intraplaque neovascularization in recent and future ischemic stroke in patients with mild carotid stenosis
Luni ZHANG ; Rong WU ; Jing CHEN ; Shiyao GU ; Caixia JIA
Ultrasonography 2025;44(1):62-71
Purpose:
There is still insufficient evidence for predicting stroke risk in patients with mild carotid atherosclerotic stenosis. This study aimed to explore the association between carotid intraplaque neovascularization (IPN) in mild stenosis and ischemic stroke, using contrast-enhanced ultrasound (CEUS) imaging.
Methods:
This retrospective observational study included 369 patients from July 2021 to March 2022. These patients were categorized as symptomatic or asymptomatic based on their recent history of ipsilateral ischemic stroke. Initial parameters of carotid plaques, such as IPN grading and contrast enhancement index, were assessed using B-mode ultrasonography and CEUS. The follow-up period lasted 12 months or until a newly-developed ischemic stroke occurred. Logistic regression models and Cox proportional-hazards regression models were employed to explore the associations between ultrasonic parameters and the incidence of recent and future ischemic strokes.
Results:
In patients with mild stenosis, both increasing age and grade 2 carotid IPN were significant predictors of recent primary ischemic stroke. Furthermore, grade 2 carotid IPN independently predicted future ischemic strokes in both symptomatic and asymptomatic patients.
Conclusion
This study demonstrated that carotid IPN as detected by CEUS imaging holds potential as a useful non-invasive biomarker for predicting recent and future ischemic strokes in patients with mild carotid stenosis.
5.The role of intraplaque neovascularization in recent and future ischemic stroke in patients with mild carotid stenosis
Luni ZHANG ; Rong WU ; Jing CHEN ; Shiyao GU ; Caixia JIA
Ultrasonography 2025;44(1):62-71
Purpose:
There is still insufficient evidence for predicting stroke risk in patients with mild carotid atherosclerotic stenosis. This study aimed to explore the association between carotid intraplaque neovascularization (IPN) in mild stenosis and ischemic stroke, using contrast-enhanced ultrasound (CEUS) imaging.
Methods:
This retrospective observational study included 369 patients from July 2021 to March 2022. These patients were categorized as symptomatic or asymptomatic based on their recent history of ipsilateral ischemic stroke. Initial parameters of carotid plaques, such as IPN grading and contrast enhancement index, were assessed using B-mode ultrasonography and CEUS. The follow-up period lasted 12 months or until a newly-developed ischemic stroke occurred. Logistic regression models and Cox proportional-hazards regression models were employed to explore the associations between ultrasonic parameters and the incidence of recent and future ischemic strokes.
Results:
In patients with mild stenosis, both increasing age and grade 2 carotid IPN were significant predictors of recent primary ischemic stroke. Furthermore, grade 2 carotid IPN independently predicted future ischemic strokes in both symptomatic and asymptomatic patients.
Conclusion
This study demonstrated that carotid IPN as detected by CEUS imaging holds potential as a useful non-invasive biomarker for predicting recent and future ischemic strokes in patients with mild carotid stenosis.
6.The role of intraplaque neovascularization in recent and future ischemic stroke in patients with mild carotid stenosis
Luni ZHANG ; Rong WU ; Jing CHEN ; Shiyao GU ; Caixia JIA
Ultrasonography 2025;44(1):62-71
Purpose:
There is still insufficient evidence for predicting stroke risk in patients with mild carotid atherosclerotic stenosis. This study aimed to explore the association between carotid intraplaque neovascularization (IPN) in mild stenosis and ischemic stroke, using contrast-enhanced ultrasound (CEUS) imaging.
Methods:
This retrospective observational study included 369 patients from July 2021 to March 2022. These patients were categorized as symptomatic or asymptomatic based on their recent history of ipsilateral ischemic stroke. Initial parameters of carotid plaques, such as IPN grading and contrast enhancement index, were assessed using B-mode ultrasonography and CEUS. The follow-up period lasted 12 months or until a newly-developed ischemic stroke occurred. Logistic regression models and Cox proportional-hazards regression models were employed to explore the associations between ultrasonic parameters and the incidence of recent and future ischemic strokes.
Results:
In patients with mild stenosis, both increasing age and grade 2 carotid IPN were significant predictors of recent primary ischemic stroke. Furthermore, grade 2 carotid IPN independently predicted future ischemic strokes in both symptomatic and asymptomatic patients.
Conclusion
This study demonstrated that carotid IPN as detected by CEUS imaging holds potential as a useful non-invasive biomarker for predicting recent and future ischemic strokes in patients with mild carotid stenosis.
7.Comparison of Quality Change During Processing Process of Achyranthis Bidentatae Radix from Different Origins Based on Color-component Correlation Analysis
Haohan DUAN ; Hao JIA ; Jingwei LEI ; Chunjing YANG ; Caixia XIE ; Haiyan GONG ; Beibei ZHU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(5):143-151
ObjectiveTaking Achyranthis Bidentatae Radix(ABR) from different origins as samples, to quantitatively analyze the chemical composition and chromaticity of ABR with different processing degrees, and clarify the correlation and change law between color and composition in the processing process of ABR, so as to provide reference for the quality evaluation of processed products of ABR. MethodThe colorimeter is used to measure the chromaticity values of three kinds of processing degrees of ABR in different origins to show the color value change trend during the processing process, and the color parameters of wine-processed and salt-processed products of ABR with different processing degrees were analyzed by principal component analysis(PCA), orthogonal partial least squares-discriminant analysis(OPLS-DA) and other analysis methods. The contents of eight representative components of ABR were measured by high performance liquid chromatography(HPLC), the correlation between chromaticity and each representative component was analyzed by Pearson correlation analysis, and the applicability of the selected eight representative components was further verified by Fisher linear discriminant analysis, and the wine-processed and salt-processed products of ABR with different processing degrees were grouped according to the degree of processing, and 48 samples of wine-processed and salt-processed products with different processing degrees were used as training samples. Taking the contents of 5-hydroxymethylfurfural, polypodine B, β-ecdysterone, 25R-inokosterone, 25S-inokosterone, ginsenoside Ro, chikusetsusaponin Ⅳa and polysaccharides as variables, the discriminant function was established respectively, and 12 samples of wine-processed and salt-processed products of ABR with different processing degrees were back-tested to verify the discriminant function and test the reliability of the function. ResultPCA and OPLS-DA results showed that ABR samples with different processing degrees were classified into clusters, and the results could significantly distinguish different processed products. During the process of wine and salt processing, the contents of 5-hydroxymethylfurfural, ginsenoside Ro, and chikusetsusaponin Ⅳa gradually increased with the deepening of the processing degree, while the contents of polypodine B, β-ecdysterone, 25R-inokosterone, 25S-inokosterone and polysaccharides showed a gradual decreasing trend, indicating these 8 components increased and decreased to different degrees in the process of wine and salt processing. The results of Pearson correlation analysis showed that the 5-hydroxymethylfurfural content of the samples with different processing degrees of wine-processed and salt-processed products were negatively correlated with the brightness value(L*) and the total color difference value(E*ab)(P<0.01), and positively correlated with the red-green value(a*) and the yellow-blue value(b*)(P<0.01), and that the content of polypodine B and polysaccharides were positively correlated with L* and E*ab(P<0.01). The discriminant functions of wine-processed and salt-processed products of ABR were established by Fisher linear discriminant analysis, and their accuracy rates in the training samples were 93.75% and 95.83%, respectively. Twelve test samples of wine-processed and salt-processed products with different processing degree were back substitution, and the correct rate was 100%. ConclusionThe trend of composition and color changes of ABR with different processing degrees in different production areas is relatively consistent, and the color value can better distinguish ABR with different processing degrees, and the color of ABR is related to some representative components in the processing process, indicating that the color can provide reference for the identification of the processing degree of ABR and the prediction of component content.
8.Associations between carotid plaque shape, biomechanical parameters, and ischemic stroke in mild carotid stenosis with a single plaque
Man ZHAO ; Luni ZHANG ; Jing CHEN ; Shiyao GU ; Rong WU ; Caixia JIA
Ultrasonography 2024;43(3):209-219
Purpose:
This cross-sectional cohort-comparison observational study investigated the value of high-frame-rate vector flow (V Flow) imaging for evaluating differences in carotid plaque shape and biomechanical parameters in patients with mild stenosis according to a recent history of ipsilateral ischemic stroke.
Methods:
The present study included 352 patients from February 2023 to October 2023, who were categorized as symptomatic or asymptomatic based on a history of recent ischemic stroke and ipsilateral ischemic lesions detected on head computed tomography or magnetic resonance imaging. A Mindray Resona R9 system was used for B-mode ultrasonography and V Flow imaging. The upstream and downstream surfaces of the plaques were examined at the carotid bifurcation for wall shear stress (WSS), oscillatory shear index (OSI), and turbulence index, which performed peri-plaque biomechanical condition. Multivariable logistic regression models were used to determine associations between plaque shape, V Flow parameters, and ischemic stroke.
Results:
Symptomatic patients exhibited higher WSS values for the upstream and downstream surfaces of carotid plaque, as well as higher OSI and turbulence index values for the downstream surface. Type Ⅲ plaques and higher WSS and OSI values for the downstream surface of the plaque were significantly associated with ischemic stroke. Type Ⅲ plaques were more prevalent in symptomatic patients and demonstrated much higher WSS and OSI values for the downstream plaque surface in both groups.
Conclusion
High-frame-rate V Flow imaging could assess peri-plaque biomechanical forces and may provide effective imaging biomarkers for early prediction of ischemic stroke in patients with mild stenosis.
9.Associations between carotid plaque shape, biomechanical parameters, and ischemic stroke in mild carotid stenosis with a single plaque
Man ZHAO ; Luni ZHANG ; Jing CHEN ; Shiyao GU ; Rong WU ; Caixia JIA
Ultrasonography 2024;43(3):209-219
Purpose:
This cross-sectional cohort-comparison observational study investigated the value of high-frame-rate vector flow (V Flow) imaging for evaluating differences in carotid plaque shape and biomechanical parameters in patients with mild stenosis according to a recent history of ipsilateral ischemic stroke.
Methods:
The present study included 352 patients from February 2023 to October 2023, who were categorized as symptomatic or asymptomatic based on a history of recent ischemic stroke and ipsilateral ischemic lesions detected on head computed tomography or magnetic resonance imaging. A Mindray Resona R9 system was used for B-mode ultrasonography and V Flow imaging. The upstream and downstream surfaces of the plaques were examined at the carotid bifurcation for wall shear stress (WSS), oscillatory shear index (OSI), and turbulence index, which performed peri-plaque biomechanical condition. Multivariable logistic regression models were used to determine associations between plaque shape, V Flow parameters, and ischemic stroke.
Results:
Symptomatic patients exhibited higher WSS values for the upstream and downstream surfaces of carotid plaque, as well as higher OSI and turbulence index values for the downstream surface. Type Ⅲ plaques and higher WSS and OSI values for the downstream surface of the plaque were significantly associated with ischemic stroke. Type Ⅲ plaques were more prevalent in symptomatic patients and demonstrated much higher WSS and OSI values for the downstream plaque surface in both groups.
Conclusion
High-frame-rate V Flow imaging could assess peri-plaque biomechanical forces and may provide effective imaging biomarkers for early prediction of ischemic stroke in patients with mild stenosis.
10.Associations between carotid plaque shape, biomechanical parameters, and ischemic stroke in mild carotid stenosis with a single plaque
Man ZHAO ; Luni ZHANG ; Jing CHEN ; Shiyao GU ; Rong WU ; Caixia JIA
Ultrasonography 2024;43(3):209-219
Purpose:
This cross-sectional cohort-comparison observational study investigated the value of high-frame-rate vector flow (V Flow) imaging for evaluating differences in carotid plaque shape and biomechanical parameters in patients with mild stenosis according to a recent history of ipsilateral ischemic stroke.
Methods:
The present study included 352 patients from February 2023 to October 2023, who were categorized as symptomatic or asymptomatic based on a history of recent ischemic stroke and ipsilateral ischemic lesions detected on head computed tomography or magnetic resonance imaging. A Mindray Resona R9 system was used for B-mode ultrasonography and V Flow imaging. The upstream and downstream surfaces of the plaques were examined at the carotid bifurcation for wall shear stress (WSS), oscillatory shear index (OSI), and turbulence index, which performed peri-plaque biomechanical condition. Multivariable logistic regression models were used to determine associations between plaque shape, V Flow parameters, and ischemic stroke.
Results:
Symptomatic patients exhibited higher WSS values for the upstream and downstream surfaces of carotid plaque, as well as higher OSI and turbulence index values for the downstream surface. Type Ⅲ plaques and higher WSS and OSI values for the downstream surface of the plaque were significantly associated with ischemic stroke. Type Ⅲ plaques were more prevalent in symptomatic patients and demonstrated much higher WSS and OSI values for the downstream plaque surface in both groups.
Conclusion
High-frame-rate V Flow imaging could assess peri-plaque biomechanical forces and may provide effective imaging biomarkers for early prediction of ischemic stroke in patients with mild stenosis.

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