1.Efficient synthesis of polydatin by a two-enzyme coupled with one-pot method.
Jingli DAI ; Zixu YAN ; Kexue ZHAO ; Xiaoli LI ; Yongjun ZANG ; Qilin XU ; Fucheng ZHU
Chinese Journal of Biotechnology 2025;41(1):461-473
Traditional Chinese medicine of Polygonum cuspidatum has been utilized in China for thousands of years. Its primary active compound, polydatin, exhibits a variety of pharmacological effects including the regulation of glucose and lipid metabolism, suppression of cough and asthma, as well as antibacterial and anti-inflammatory properties. However, conventional methods for polydatin production are inadequate to satisfy the market demand. This study aims to explore the green and efficient preparation of polydatin. With resveratrol as the substrate, we efficiently synthesized polydatin by using the triple mutant IGW (Y14I/I62G/M315W) of the glycosyltransferase UGTBS based on a strategy of two-enzyme coupled with one-pot and realized the recycling of uridine diphosphate-glucose (UDPG). The conditions of the two-enzyme reaction were optimized. Under the conditions of 35 ℃, pH 8.0, IGW: AtSuSy1 activity ratio of 3:4, dimethyl sulfoxide (DMSO) volume fraction of 5%, uridine diphosphate (UDP) concentration of 0.10 mmol/L, and sucrose concentration of 0.6 mol/L, the conversion of 2 mmol/L resveratrol reached 80.6% within 1 h, and the proportion of polydatin was over 90%. This study achieved the recycling of UDPG via a two-enzyme coupling system and shortened the reaction time. At the same time, the fed-batch strategy was adopted, and the yield of polydatin reached 6.28 g/L after 24 h in the one-pot coupling reaction, which provided a new strategy for green and efficient preparation of polydatin.
Stilbenes/chemistry*
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Glucosides/biosynthesis*
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Resveratrol
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Fallopia japonica/chemistry*
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Glycosyltransferases/genetics*
2.Evaluation of flavonoids in Chimonanthus praecox based on metabolomics and network pharmacology.
Dan ZHOU ; Yanbei ZHAO ; Zixu WANG ; Qingwei LI
Chinese Journal of Biotechnology 2025;41(2):602-617
Flavonoids are key bioactive components for evaluating the pharmacological activities of Chimonanthus praecox. Exploring the potential flavonoids and pharmacological mechanisms of C. praecox lays a foundation for the rational development and efficient utilization of this plant. This study employed ultra-performance liquid chromatography-tandem mass spectrometry-based widely targeted metabolomics to comprehensively identify the flavonoids in C. praecox. Network pharmacology was employed to explore the bioactive flavonoids and their mechanisms of action. Molecular docking was adopted to validate the predicted results. Finally, the content of bioactive flavonoids in different varieties of C. praecox was measured. The widely targeted metabolomics analysis identified 387 flavonoids in C. praecox, and the flavonoids varied among different varieties. Network pharmacology predicted 96 chemical components including 19 bioactive compounds, 181 corresponding targets and 2 504 disease targets, among which 99 targets were shared by the active components and the disease. Thirty-three core targets were predicted, involving 229 gene ontology terms and 99 pathways (P≤0.05), which indicated that the flavonoids components of C. praecox exhibited pharmacological activities including antioxidant, anti-inflammatory, antimicrobial, and antiviral activities. Topological analysis screened out five core components (salvigenin, laricitrin, isorhamnetin, quercetin, and 6-hydroxyluteolin) and five core targets (SRC, PIK3R1, AKT1, ESR1, and AKR1C3). The predicted bioactive flavonoids from C. praecox stably bound to key targets, which indicated that these flavonoids possessed potential bioactivities in their interactions with the targets. The flavonoids in C. praecox exerted pharmacological activities in a multi-component, multi-target, and multi-pathway manner. The combined application of metabolomics and network pharmacology provides a theoretical basis for in-depth studies on the pharmacological effects and mechanisms of C. praecox.
Flavonoids/metabolism*
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Network Pharmacology
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Metabolomics/methods*
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Molecular Docking Simulation
;
Calycanthaceae/chemistry*
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Tandem Mass Spectrometry
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Drugs, Chinese Herbal/chemistry*
3.Genetic diversity and molecular identity of Prunus mume with both ornamental and edible values based on fluorescence-labeled simple sequence repeat (SSR) markers.
Zixu WANG ; Dan ZHOU ; Yanbei ZHAO ; Yuhang TONG ; Weijun ZHENG ; Qingwei LI
Chinese Journal of Biotechnology 2025;41(2):639-656
We studied the genetic diversity and established the DNA molecular identify for Prunus mume with both ornamental and edible values, aiming to collect, identify, evaluate, and breed new varities of this plant and promote the upgrading of the P. mume industry chain in northern China. We employed 13 pairs of primers with good polymorphism, clear bands, and good repeatability to analyze the genetic diversity and establish the molecular identify of 68 germplasm accessions of P. mume with both ornamental and edible values from Xingtai, Hebei Province. We then employed the unweighted pair-group method with arithmetic means (UPGMA) to perform the cluster analysis based on genetic distance. After that, we analyzed the genetic structure of the 68 germplasm accessions based on a Bayesian model. The 13 pairs of SSR primers amplified a total of 124 alleles from 68 P. mume germplasm accessions, with the mean number of alleles (Na) of 9.538 5, the minor allele frequency (MAF) of 0.369 3, the mean number of effective alleles (Ne) of 4.483 5, and the mean Shannon genetic diversity index (I) of 1.712 4. The mean Nei's gene diversity index (H) of 0.763 7, the mean observed heterozygosity (Ho) of 0.719 5, the mean expected heterozygosity (He) of 0.769 3, the mean polymorphism information content (PIC) of 0.733 6, and the mean genetic similarity (GS) of 0.772 9 suggested that there were significant genetic differences and rich genetic diversity among the studied P. mume germplasm accessions. The cluster analysis revealed that the 68 accessions were classified into three groups, with the mean genetic distance of 0.622 6. The population structure analysis classified the germplasm accessions into two populations. According to the PIC of primers, we selected primers for combination and constructed the combination with the fewest primers required for germplasm differentiation of P. mume with both ornamental and edible values. This study provides a theoretical basis for the innovation and industrial upgrading of P. mume with both ornamental and edible values in gardening and the improvement of breeding efficiency.
Prunus/classification*
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Microsatellite Repeats/genetics*
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Genetic Variation
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China
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Phylogeny
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Polymorphism, Genetic
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DNA, Plant/genetics*
;
Alleles
4.Construction of the clinical practice pathway for narrative medicine in traditional Chinese medicine
Zixu WANG ; Jingjing ZHAO ; Yipin LIU ; Jie LI ; Qiuli YANG ; Yongyan WANG
Chinese Medical Ethics 2024;37(11):1251-1262
Objective:To construct the clinical practice pathway for narrative medicine in traditional Chinese medicine(TCM),with a view to providing clinical practice guidelines for narrative medicine in TCM for frontline practitioners.Methods:Using the realistic literature review and the Nominal Group Technique(NGT),the paper systematically sorted out the practices of humanistic care in ancient Chinese medical books and famous medical cases,as well as constructed the first draft of the clinical practice pathway and details for narrative medicine in TCM.Subsequently,experts from multiple fields were invited to demonstrate by using the NGT.After in-depth discussion and collective voting,various operational modules,and their detailed rules and supporting tools were determined,thus completing the construction of the entire practical pathway.Results:A complete set of clinical practice pathways for narrative medicine in TCM had been established.It encompassed six core modules,including"start of diagnosis and treatment","communication of disease conditions","diagnosis and explanation","joint decision-making","end of diagnosis and treatment",and"reflection and summary".Besides,detailed operating rules and supporting tools were also provided.Conclusion:The clinical practice pathway for narrative medicine in TCM integrates the humanistic spirit of TCM,the core concepts of narrative medicine,and the communication skills of psychology,providing medical workers with standardized,procedural,and operationally flexible practice guidance,which helps both doctors and patients to better communicate,empathize,and make joint decisions throughout the entire process of diagnosis and treatment.
5.Recent advance in oxidative stress after intracranial hemorrhage
Xiaoqi YANG ; Jianlin DING ; Zhong WANG ; Yijiang LI ; Junchi WANG ; Xuehai DENG ; Zixu WANG ; Yiqian CHEN ; Long ZHAO
Chinese Journal of Neuromedicine 2024;23(10):1043-1049
Intracerebral hemorrhage (ICH) is a hemorrhagic cerebrovascular disease with high incidence and mortality. Oxidative stress response plays an important role in the pathological and physiological processes of ICH, and is also a potential effective target for clinical treatment. In this paper, the pathogenesis of oxidative stress after ICH, mechanism of nerve and vascular injury in oxidative stress, and detection and treatment of oxidative stress are reviewed in order to provide references for basic research and clinical practice in ICH.
6.Qualitative Research on the Influence of Narrative Medicine Practice on the Professional Identity of Postgraduates Majoring in Traditional Chinese Medicine
Jingjing ZHAO ; Qiuli YANG ; Zixu WANG
Chinese Medical Ethics 2023;36(11):1191-1199
【Objective:】 To explore the practice experience of narrative medicine among postgraduates majoring in Traditional Chinese Medicine (TCM) and its impact on their professional identity. 【Methods:】 A total of 56 first-year master’s and doctoral degree students majoring in TCM at a university were selected through purposive sampling for a 16-hour narrative medicine course training and 4 weeks of clinical practice in narrative medicine. They were asked to complete an open-ended writing assignment on their practical experience after this clinical practice. Subsequently, the topic analysis method in qualitative research was used to analyze their writing contents. 【Results:】 After learning narrative medicine, the students’ practical experience in clinical practice included 3 major themes and 9 sub-themes: wide application range (all diagnosis and treatment stages, multiple departments, and disease types), integration of multiple communication skills and diagnosis and treatment methods (communication skills, psychological techniques, reflection, and TCM humanistic diagnosis and treatment methods), and increasing professional identity (achieving mutual healing, building the harmonious doctor-patient relationship, and enhancing confidence in TCM). The themes were further integrated to form the relationship between the narrative medical practice cycle and professional identity, and presented in the form of charts. 【Conclusion:】 By exploring the narrative medical practice experience among TCM postgraduates, it was found that through the use of multiple methods and techniques in the entire diagnosis and treatment stage and multi-disciplinary diseases, it is possible to harmonize the doctor-patient relationship, achieve mutual healing, and enhance confidence in TCM. These internalized recognition and sense of achievement contributes to improving professional identity, which may provide new ideas for the development of professional identity for medical students in China and the exploration of the localization of narrative medical practice.
7.Effect and mechanism of salidroside for 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine induced Parkinson's disease in mice
Zixu SUN ; Xia DONG ; Baomin ZHAO ; Haili JIA ; Xiaorui MA ; Wei GU
Journal of Clinical Medicine in Practice 2023;27(22):55-61,66
Objective To investigate the effect and mechanism of salidroside on1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)-induced Parkinson's disease(PD)in mice.Methods C57BL/6J mice were induced by MPTP to establish a PD mouse model and the movement track of mice was re-corded by open field experiment.The expression of tyrosine hydroxylase(TH),α-synuclein(α-syn),nuclear factor E2 associated factor 2(Nrf2)and quinone oxidoreductase 1(NQO1)were detected by immunohistochemistry in brain tissue of mice.SN4741 cells were stimulated by MPTP to establish a PD cell model in vitro.MPTP stimulation of SN4741 cells to establish an in vitro cell mod-el of PD.After pretreatment with salidroside,TUNEL was used to detect cell apoptosis in each group.The expressions of TH,α-syn,Nrf2 and NQO1 were detected by immunofluorescence.Nrf2 expres-sion was knocked down by being transfected with si-RNA,and apoptosis condition was detected by TUNEL assay.The expression levels of interleukin-1β(IL-1β)and interleukin-18(IL-18)protein in brain tissue extract and cell culture were detected by enzyme-linked immunosorbent assay(ELISA).Results The open field experiment results showed that salidroside could improve 3 min total distance of activity,average speed and activity track of PD mice(P<0.05).Compared with the PD model mice,the salidroside treated mice showed significantly increased expression of TH,Nrf2 and NQO1 in the brain,and reduced expression of IL-18,α-Syn,IL-1β.Compared with the MPTP stimulated cells,the salidroside pretreatment group showed a decrease in cell apoptosis and an increase in the expressions of TH,Nrf2 and NQO1,but reduced expressions in α-Syn,IL-1β and IL-18.After si-RNA knockdown of Nrf2 expression,the protective effect of salidroside on MPTP stimulated cells weakened or even disappeared.Conclusion Salidroside may alleviate neuronal apoptosis by activating Nrf2-ARE signaling pathway,which is expected to be a new drug for PD treatment.
8.The effect of vitrectomy in the treatment of different types of chorioretinal coloboma with retinal detachment
Jiao DU ; Xiaoli LI ; Zun ZHAO ; Dongdong WANG ; Handong DAN ; Pingling SHI ; Zixu HUANG ; Suhan WANG ; Qiongqiong YUAN ; Zongming SONG
Chinese Journal of Ocular Fundus Diseases 2023;39(12):969-973
Objective:To observe the efficacy of pars plana vitrectomy (PPV) in the treatment of different types of chorioretinal coloboma with retinal detachment (RD).Methods:A single-center, retrospective clinical study. From April 2021 to March 2023, 24 eyes of 23 patients who were diagnosed as chorioretinal coloboma with RD in Henan Provincial Eye Hospital were included in this study. There were 11 males with 12 eyes and 12 females with 12 eyes. The mean age was (33.3±13.7) years old. Best corrected visual acuity (BCVA), spectral domain optical coherence tomography were performed. The BCVA examination was performed using a international standard logarithmic visual acuity chart, which was converted into logarithm of the minimum angle of resolution (logMAR) visual acuity during statistics. According to the types of chorioretinal coloboma, the affected eyes were divided into the coloboma involved the optic disc group and the coloboma not involved the optic disc group, with 15 eyes and 9 eyes. According to whether the RD containing the coloboma area, the affected eyes were divided into RD containing the coloboma area group and the RD not containing the coloboma area group, with 15 eyes and 9 eyes. All eyes underwent standard pars plana three-channel 25G PPV, retinal laser photocoagulation combined with silicone oil tamponade. The follow-up time after surgery was (19.5±16.3) months. The last follow-up was the time point for efficacy determination. The retinal reattachment, BCVA recovery and postoperative complications were observed. Paired t-test or t test was performed for comparison of quantitative data. Fisher's exact test was performed for comparison of qualitative data. Results:At the last follow-up, retinal reattachment was achieved in 20 eyes (83.3%, 20/24). The logMAR BCVA of the coloboma involved the optic disc group before and after surgery were 1.85±0.62 and 1.71±0.71, the difference was no significant ( t=0.845 , P=0.412). The logMAR BCVA of the coloboma not involved the optic disc group before and after surgery were 1.75±0.45 and 0.84±0.26, the difference was statistically significant ( t=6.153 , P<0.001). The improvement of BCVA in the coloboma not involved the optic disc group was significantly higher than that in the coloboma involved the optic disc group after surgery, with statistically significant differences ( t=3.024 , P=0.006). There was no significant difference in the retinal reattachment rate between the two groups ( P=0.615). There was no significant difference in the retinal reattachment rate between the RD containing the coloboma area group and the RD not containing the coloboma area group ( P=0.259). Postoperative complications included elevated intraocular pressure in five eyes, cataract progression in ten eyes, recurrent RD in two eyes, bullous keratopathy in one eye and band-shaped keratopathy in one eye. Conclusion:PPV combined with silicone oil tamponade is safe and effective in the treatment of chorioretinal coloboma with RD, the improvement of visual acuity in the coloboma not involved the optic disc group is better than that in the coloboma involved the optic disc group after surgery.
9.Effect and mechanism of salidroside for 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine induced Parkinson's disease in mice
Zixu SUN ; Xia DONG ; Baomin ZHAO ; Haili JIA ; Xiaorui MA ; Wei GU
Journal of Clinical Medicine in Practice 2023;27(22):55-61,66
Objective To investigate the effect and mechanism of salidroside on1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)-induced Parkinson's disease(PD)in mice.Methods C57BL/6J mice were induced by MPTP to establish a PD mouse model and the movement track of mice was re-corded by open field experiment.The expression of tyrosine hydroxylase(TH),α-synuclein(α-syn),nuclear factor E2 associated factor 2(Nrf2)and quinone oxidoreductase 1(NQO1)were detected by immunohistochemistry in brain tissue of mice.SN4741 cells were stimulated by MPTP to establish a PD cell model in vitro.MPTP stimulation of SN4741 cells to establish an in vitro cell mod-el of PD.After pretreatment with salidroside,TUNEL was used to detect cell apoptosis in each group.The expressions of TH,α-syn,Nrf2 and NQO1 were detected by immunofluorescence.Nrf2 expres-sion was knocked down by being transfected with si-RNA,and apoptosis condition was detected by TUNEL assay.The expression levels of interleukin-1β(IL-1β)and interleukin-18(IL-18)protein in brain tissue extract and cell culture were detected by enzyme-linked immunosorbent assay(ELISA).Results The open field experiment results showed that salidroside could improve 3 min total distance of activity,average speed and activity track of PD mice(P<0.05).Compared with the PD model mice,the salidroside treated mice showed significantly increased expression of TH,Nrf2 and NQO1 in the brain,and reduced expression of IL-18,α-Syn,IL-1β.Compared with the MPTP stimulated cells,the salidroside pretreatment group showed a decrease in cell apoptosis and an increase in the expressions of TH,Nrf2 and NQO1,but reduced expressions in α-Syn,IL-1β and IL-18.After si-RNA knockdown of Nrf2 expression,the protective effect of salidroside on MPTP stimulated cells weakened or even disappeared.Conclusion Salidroside may alleviate neuronal apoptosis by activating Nrf2-ARE signaling pathway,which is expected to be a new drug for PD treatment.
10.Efficacy and safety of endovascular treatment vs medical treatment in anterior circulation stroke beyond 6 Hours: A systematic review and metaanalysis
Zixu Zhao ; Xin Jiang ; Ying Zhang ; Zixiao Yin ; Guohui Lu ; Yang Wang ; Michael Hall ; Lingfeng Lai
Neurology Asia 2020;25(4):439-446
Background & Objective: Endovascular treatment is the widely accepted treatment for patients with
anterior circulation stroke within 6 hours of onset of stroke. We aimed to evaluate the advantages
of endovascular treatment compared to standard medical treatment in treating patients with anterior
circulation stroke beyond the 6-hour therapeutic window. Methods: We reviewed the literature
concerning endovascular treatment versus medical treatment beyond the 6-hour therapeutic window.
Using random-effects meta-analysis, we evaluated the following outcomes: modified Rankin scale in
the three-month follow-up [excellent outcome (mRS≤1), functional independence (mRS≤2), moderate
outcome(mRS≤3)], recanalization rate at 24 hours, mortality at 90 days or in-hospital, symptomatic
intracranial hemorrhage, parenchymal hematoma type 2 and hemorrhagic infarction 1. Results: Four
studies including 642 patients were evaluated. Endovascular treatment was associated with higher
odds of excellent outcome (OR 2.55; 95% CI 1.48 to 4.41,), functional independence (OR 3.64; 95%
CI 2.43 to 5.45), moderate outcome (OR 2.70; 95% CI 1.95-3.74) and recanalization rate at 24 hours
(OR 8.81; 95%CI 2.81 to 27.69) compared to MT. No difference in the rates of mortality, symptomatic
intracranial hemorrhage, parenchymal hematoma type 2 or hemorrhagic infarction 1 was found between the 2 groups. Studies using strict perfusion imaging inclusion selection showed better moderate outcome in comparison to the studies without perfusion imaging inclusion selection (P <0.012).
Conclusion: Our study highlights the superiority of endovascular treatment over standard medical
treatment alone for treating patients with anterior circulation stroke beyond 6 hours since stroke onset,
although more studies are required for further investigation. Standard of strict selection for eligible
patients before endovascular treatment should be based on DAWN or DEFFUSE 3 inclusion criteria.


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