1.A review on the screening methods for the discovery of natural antimicrobial peptides.
Bin YANG ; Hongyan YANG ; Jianlong LIANG ; Jiarou CHEN ; Chunhua WANG ; Yuanyuan WANG ; Jincai WANG ; Wenhui LUO ; Tao DENG ; Jialiang GUO
Journal of Pharmaceutical Analysis 2025;15(1):101046-101046
Natural antimicrobial peptides (AMPs) are promising candidates for the development of a new generation of antimicrobials to combat antibiotic-resistant pathogens. They have found extensive applications in the fields of medicine, food, and agriculture. However, efficiently screening AMPs from natural sources poses several challenges, including low efficiency and high antibiotic resistance. This review focuses on the action mechanisms of AMPs, both through membrane and non-membrane routes. We thoroughly examine various highly efficient AMP screening methods, including whole-bacterial adsorption binding, cell membrane chromatography (CMC), phospholipid membrane chromatography binding, membrane-mediated capillary electrophoresis (CE), colorimetric assays, thin layer chromatography (TLC), fluorescence-based screening, genetic sequencing-based analysis, computational mining of AMP databases, and virtual screening methods. Additionally, we discuss potential developmental applications for enhancing the efficiency of AMP discovery. This review provides a comprehensive framework for identifying AMPs within complex natural product systems.
2.Cloning and expression analysis of the laccase gene RcLAC15 from Rosa chinensis.
Qi LI ; Yifang PENG ; Qijing DONG ; Qian YANG ; Xiaoyu LIU ; Yu HAN
Chinese Journal of Biotechnology 2025;41(2):845-856
Laccases (LACs), belonging to the multicopper oxidase family, are closely associated with various biological functions including lignin synthesis and responses to biotic and abiotic stresses in plants. However, few studies have reported the laccase genes in China rose (Rosa chinensis). Prickles cause difficulties to the management and harvest of R. chinensis and have become a trait concerned in the breeding. To investigate the expression patterns of laccase genes in roses, we cloned a laccase gene from an ancient variety R. chinensis 'Old Blush' and named it RcLAC15. The expression level of RcLAC15 in prickles was significantly higher than those in roots, stems, and leaves. Fifty-eight laccase genes were identified in the genome of R. chinensis, and bioinformatics analysis revealed that RcLAC15 was a homolog of AtLAC15, predicting that RcLAC15 was a stable hydrophilic protein without transmembrane structures. The recombinant expression vector pBI121-proRcLAC15:: GUS was introduced into Arabidopsis, and GUS staining results showed that the RcLAC15 promoter specifically drove GUS gene expression at the edges of Arabidopsis leaves. In summary, RcLAC15 is a gene specifically expressed in the prickles of R. chinensis. This discovery provides a reference for exploring the biological functions of laccase genes in the prickles of R. chinensis.
Laccase/metabolism*
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Rosa/enzymology*
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Cloning, Molecular
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Gene Expression Regulation, Plant
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Plant Proteins/metabolism*
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Arabidopsis/metabolism*
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Plants, Genetically Modified/metabolism*
3.Professor WANG Xixing's Experience in Differentiating the Treatment of Brain Tumor from Emotions and Minds
Xinyue WANG ; Yuankun HAN ; Lanzhi ZHANG ; Yifang LI ; Xi YANG
Journal of Traditional Chinese Medicine 2024;65(21):2184-2188
To summarise Professor WANG Xixing's experience in treating brain tumour from emotions and minds. Professor WANG regarded abnormal emotions and minds as the key to the treatment of brain tumor. Emotions and minds internal damage lead to malfunction of the five zang organs, resulting in phlegm, stasis, dampness, toxicity and other pathogenic qi gathering in the brain and developing into brain tumour. The treatment advocated regulating qi of the five zang organs and dispelling phlegm, blood stasis, dampness and toxins at the same time. The brain tumour is classified into four syndromes: heart-spleen deficiency, liver depression and spleen deficiency, failure of the heart and kidney to interact, and liver-kidney yin deficiency, with Guipi Yangshen Decoction (归脾养神汤) to fortify the spleen and nourish the heart, Guishao Shunzhi Decoction (归芍顺志汤) to soothe the liver and fortify the spleen, Liangui Tongshen Decoction (连桂通神汤) to restore interaction between the heart and the kidney, and Zishui Qingmu Huashen Decoction (滋水清木化神汤) to nourish the kidney and clear liver. At the same time, it was emphasised that the patient's emotional changes should be paid attention to, and psychological guidance should be given at the right time, so as to overcome the disease with emotions and mind.
4.Tongxie Yaofang Regulates Expression of NKG2DL to Enhance Anti-tumor Effect of NK Cells in Colon Cancer under Chronic Stress
Yan'e HU ; Yuqing HUANG ; Yi YANG ; Yifang JIANG ; Xi FU ; Fengming YOU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(1):103-111
ObjectiveTo observe the effect of Tongxie Yaofang on the function of tumor-related natural killer (NK) cells under chronic stress and explore the possible molecular mechanism. MethodFifty SPF-grade BABL/C male mice were randomized into normal, model, and low-, medium-, and high-dose (6.825, 13.65, and 27.3 g·kg-1, respectively) Tongxie Yaofang groups, with 10 mice in each group. Other groups except the blank group were subjected to 7 days of chronic restraint stress, and then forced swimming and tail suspension tests were carried out to evaluate the modeling performance. After the successful modeling, rats in Tongxie Yaofang groups were administrated with low-, medium-, and high-doses of Tongxie Yaofang by gavage, while those in the other groups were administrated with normal saline by gavage. After 14 days, each group of mice was inoculated with subcutaneous colon cancer to establish the model of colon cancer under chronic stress. The pathological changes of the tumor tissue in each group of mice were observed using hematoxylin-eosin (HE) staining. The content of CD49b-positive cells in the peripheral blood and tumor tissue of mice was measured by flow cytometry. Enzyme-linked immunosorbent assay (ELISA) was employed to measure the content of molecules associated with NK cell activation in the peripheral blood. Western blot was employed to determine the protein levels of major histocompatibility complex class Ⅰ polypeptide-related sequences A and B (MICA+MICB) and UL-16-binding protein 1 (ULBP1) in the tumor tissue. ResultCompared with the normal group, the model group showed a decrease in 5-hydroxytryptamine (5-HT) content and an increase in corticosterone (CORT) content in the serum (P<0.05). Compared with the model group, Tongxie Yaofang increased the 5-HT content and decreased the CORT content (P<0.05, P<0.01). Compared with the normal group, the modeling increased the tumor volume and weight (P<0.05), while Tongxie Yaofang inhibited such increases with no statistical significance. The tumor cells in the model group presented neat arrangement, irregular shape, uneven size, obvious atypia, common nuclear division, and small necrotic area, and blood vessels were abundant surrounding the tumor cells. Compared with the model group, Tongxie Yaofang groups showed sparse arrangement of tumor cells, different degrees of patchy necrosis areas in the tumor, and karyorrhexis, dissolution, and nuclear debris in the necrotic part. Compared with the normal group, the model group showed reduced CD49b-positive cells in the peripheral blood and tumor tissue (P<0.01). Compared with the model group, Tongxie Yaofang increased CD49b-positive cells (medium dose P<0.01, high dose P<0.05, P<0.01). Compared with the normal group, the modeling lowered the serum levels of granzymes-B (Gzms-B), perforin (PF), interferon (IFN)-γ, and tumor necrosis factor (TNF)-α (P<0.05, P<0.01). Compared with the model group, low-dose Tongxie Yaofang elevated the serum levels of PF, Gzms-B, and TNF-α (P<0.05, P<0.01), and medium-dose Tongxie Yaofang elevated the serum levels of Gzms-B, PF, IFN-γ, and TNF-α (P<0.05, P<0.01). In addition, high-dose Tongxie Yaofang elevated the serum levels of PF, IFN-γ, and TNF-α (P<0.01). Compared with the normal group, the model group presented down-regulated protein level of ULBP1 (P<0.05). Compared with the model group, low-, medium-, and high-dose Tongxie Yaofang up-regulated the protein level of ULBP1 (P<0.05, P<0.01), and medium- and high-dose Tongxie Yaofang up-regulated the protein level of MICA+MICB (P<0.05, P<0.01). ConclusionTongxie Yaofang may promote NK cell activation by up-regulating the expression of MICA+MICB and ULBP1, thereby delaying the progression of colon cancer under chronic stress.
5.Prognosis of different hemodynamic classifications in patients with pulmonary hypertension due to left heart disease
Yuan TANG ; Yanping SHI ; Lu CHEN ; Yifang SUO ; Shengen LIAO ; Cheang LOKFAI ; Yanli ZHOU ; Rongrong GAO ; Jing SHI ; Wei SUN ; Hao ZHANG ; Yanhui SHENG ; Rong YANG ; Xiangqing KONG ; Xinli LI ; Haifeng ZHANG
Chinese Journal of Cardiology 2024;52(10):1177-1185
Objective:To compare the prognostic values of different classification by using transpulmonary pressure gradient (TPG), diastolic pressure gradient (DPG) and pulmonary vascular resistance (PVR) in patients with pulmonary hypertension due to left heart disease (PH-LHD), and investigated hemodynamic and clinical factors associated with mortality in patients with PH-LHD.Methods:This was a single-center prospective cohort study. In-hospital patients diagnosed with PH-LHD via right heart catheterization at the Department of Cardiology, the First Affiliated Hospital of Nanjing Medical University, from September 2013 to December 2019 were enrolled. Patients were divided according to TPG (cutoff value 12 mmHg; 1 mmHg=0.133 kPa), DPG (cutoff value 7 mmHg), PVR (cutoff value 3 Wood Units), and the combination of TPG and PVR. Baseline characteristic was recorded. All patients were followed up until the occurrence of endpoint event, defined as all-cause death that occurred during the follow-up period, or until April 18, 2022. Receiver operating characteristic curves were used to compare the predictive value of 3 classification methods for all-cause death in PH-LHD patients. The optimal cutoff values were calculated using Jorden index. Survival analysis was performed using Kaplan-Meier analysis, and log-rank test was used to compare the predictive efficacy of classification methods based on optimal cutoff values or guidance-recommended thresholds for the survival of PH-LHD patients. Variables showing statistical significance in the univariate analysis were incorporated into multivariate Cox regression model to analyze the independent risk factors for all-cause mortality.Results:A total of 243 patients were enrolled, aged (54.9±12.7) years old, including 169 (69.5%) males. During a median follow-up of 57 months, there were 101 (41.6%) deaths occurred. Grouping results were as follows: (1) TPG: TPG≤12 mmHg group 115 patients, TPG>12 mmHg group 128 patients; (2) DPG: DPG<7 mmHg group 193 patients, DPG≥7 mmHg group 50 patients; (3) PVR: PVR≤3 Wood Units group 108 patients, PVR>3 Wood Units group 135 patients; (4) TPG and PVR: TPG≤12 mmHg and PVR≤3 Wood Units group 89 patients, TPG>12 mmHg and PVR>3 Wood Units group 109 patients. PVR ( AUC=0. 698,95% CI:0.631-0.766) had better predictive value for all-cause mortality than TPG ( AUC=0.596, 95% CI: 0.523-0.669) and DPG ( AUC=0.526, 95% CI: 0.452-0.601) (all P<0.05). The optimal cutoff values for TPG, DPG, and PVR were13.9 mmHg, 2.8 mmHg, and 3.8 Wood Units, respectively. Kaplan-Meier analysis based on the optimal cutoff values or guidance-recommended thresholds showed that PVR and TPG were the predictors of survival ( P<0.05), while DPG did not showed significance ( P>0.05). Multivariate Cox regression analysis showed that age, PVR and log 2N-terminal pro-B-type natriuretic peptide were independent risk factors for all-cause mortality in PH-LHD patients (all P<0.05). Conclusion:Classification according to PVR was most valuable in predicting all-cause death in PH-LHD patients, while TPG showed moderate predictive ability and DPG had no predictive value.
6.Correlation analysis on serum proline dehydrogenase levels and left ventricular systolic function in patients with chronic heart failure
Kaitong YANG ; Lili HE ; Qingjuan ZUO ; Xinwei YU ; Yifang GUO
Journal of Jilin University(Medicine Edition) 2024;50(6):1719-1727
Objective:To discuss the differences in serum proline dehydrogenase(ProDH)levels among chronic heart failure(CHF)patients with different ejection fraction types,and to clarify the effect of ProDH levels on cardiac function.Methods:A retrospective analysis of clinical data of 118 CHF patients was conducted.These patients were divided into heart failure with reduced ejection fraction(HFrEF)group(n=39),heart failure with mid-range ejection fraction group(HFmrEF)(n=42),and heart failure with preserved ejection fraction(HFpEF)group(n=37).A total of 45 non-CHF patients hospitalized during the same period were collected as control group.The general data of all the subjects in various groups were collected,and the levels of biochemical indicators and cardiac structure indicators in serum of all the subjects were detected.Spearman correlation analysis and point-biserial correlation analysis were used to analyze the correlation between serum ProDH levels and various biochemical indicators;multivariate Logistic regression analysis was used to analyze the factors influencing HFrEF and HFmrEF.Results:Compared with control group,the usage rate of beta-blockers of the patients in HFpEF group was significantly increased(P<0.05);in HFmrEF group,the percentage of male patients,the usage rate of statins,and the usage rate of beta-blockers were all significantly increased(P<0.05);in HFrEF group,the age and systolic blood pressure(SBP)of the patients were significantly decreased(P<0.05),while the usage rates of statins and beta-blockers of the patients were significantly increased(P<0.05).Compared with HFpEF group,the age of the patients in HFmrEF group was significantly decreased(P<0.05),and the percentage of male patients and the usage rate of statins were significantly increased(P<0.05);the age of the patients in the HFrEF group was significantly decreased(P<0.05),and the usage rate of statins was significantly increased(P<0.05).Compared with HFmrEF group,the SBP of the patients in HFrEF group was significantly decreased(P<0.05).Compared with control group,the serum levels of low-density lipoprotein cholesterol(LDL-c)of the patients in HFpEF and HFmrEF groups were significantly decreased(P<0.05),while the levels of N-terminal pro-brain natriuretic peptide(NT-proBNP)were significantly increased(P<0.05);the serum levels of glomerular filtration rate(GFR)and ProDH of the patients in HFrEF group were significantly decreased(P<0.05),and the levels of fasting blood glucose(FBG)and NT-proBNP were significantly increased(P<0.05).Compared with HFpEF group,the serum hemoglobin(Hb)level of the patients in HFmrEF group was significantly increased(P<0.05);the serum NT-proBNP level of the patients in HFrEF group was significantly increased(P<0.05),while the ProDH level was significantly decreased(P<0.05).Compared with HFmrEF group,the serum NT-proBNP level of the patients in HFrEF group was significantly increased(P<0.05).Compared with control group,the left atrial diameter(LAD)and the ratio of early diastolic mitral inflow velocity to early diastolic mitral annular velocity(E/Em)of the patients in HFpEF,HFmrEF,and HFrEF groups were significantly increased(P<0.05);the left ventricular end-diastolic diameter(LVEDD)of the patients in HFmrEF and HFrEF groups were significantly increased(P<0.05),and the left ventricular ejection fraction(LVEF)were significantly decreased(P<0.05).Compared with HFpEF group,the LVEDD of the patients in HFmrEF and HFrEF groups were significantly increased(P<0.05),and the LVEF were significantly decreased(P<0.05);the LAD of the patients In HFrEF group was significantly increased(P<0.05).Compared with HFmrEF group,the E/Em ratio of the patients in HFrEF group was significantly increased(P<0.05),and the LVEF was significantly decreased(P<0.05).The serum ProDH levels of the patients were negatively correlated with LVEDD(r=-0.210,P=0.007)and positively correlated with LVEF(r=0.220,P=0.005).Male and elevated FBG levels were the risk factors for cardiac function,while the increasing serum GFR and ProDH levels were the protective factors for cardiac function.Conclusion:There are differences in ProDH levels among the CHF patients with different ejection fraction types.The patients with poorer cardiac function have lower serum ProDH levels,and higher ProDH levels may be beneficial for improving the left ventricular systolic function in the CHF patients.
7.Concept analysis of the intrinsic capacity of the elderly
Tingting WU ; Xiaoqin WEI ; Jianhui DONG ; Tingting YANG ; Yifang YANG ; Junbo CHEN ; Xiang HE ; Yuxia MA
Chinese Journal of Nursing 2024;59(16):2037-2043
Objective To analyze and clarify the concept of intrinsic capacity of the elderly.Methods We searched studies on intrinsic capacity of the elderly from websites and databases,including PubMed,Cochrane Library,Web of Science,Embase,CINAHL,China Biomedical Literature Service System databases,CNKI,WanFang databases and VIP.We selected relevant papers from the inception of databases to July 2023 according to inclusion criteria.Rodgers evolutionary method of concept analysis was used.Results A total of 30 articles were retrieved.4 attributes were identified on intrinsic capacity,including:guided by the goal of achieving healthy aging,inherent physiological reserve capacity of individuals,rich and interactive dimensions,and a dynamic and reversible development trajectory.Its prerequisites include demographic factors,socio-economic factors,health-related charac-teristics,external environment,and other factors;post effects include early identification of declining intrinsic abilities in the elderly and timely adoption of targeted intervention measures,which are of great significance for improving the quality of life of the elderly and promoting healthy aging.The decline in intrinsic capabilities is closely related to various adverse health outcomes,posing a serious threat to the health status of the elderly.Conclusion The concept attributes of intrinsic capacity were identified by concept analysis method.In the future,research and clinical practice should be carried out based on the concept of intrinsic capacity.
8.The correlation and predictive value of GCS score,Lac,and blood transfusion in patients with hemorrhagic shock
Dongqiao YANG ; Yuan CHEN ; Xiaobin TANG ; Yifang SU
China Modern Doctor 2024;62(21):42-46
Objective To analyze the correlation between Glasgow coma score(GCS),blood lactic acid(Lac),and blood transfusion volume in traumatic emergency hemorrhagic shock patients,and their predictive value for patient prognosis.Methods Retrospective collection of clinical data from 128 trauma emergency shock patients who were treated and followed up in the Emergency Medical Center,Jinhua People's Hospital from March 2021 to May 2023.They were divided into good prognosis group(n=106)and poor prognosis group(n=22)according to their prognosis.The general information,GCS score,Lac level,and blood transfusion volume of two groups were compared.Using Cox regression model to analyze the influencing factors of prognosis in trauma emergency hemorrhagic shock patients.Establish receiver operating characteristic(ROC)curve to analyze the predictive value of GCS score,Lac level,and blood transfusion volume on the prognosis of trauma emergency hemorrhagic shock patients.Results Among 128 patients,22 had poor prognosis,accounting for 17.19%.The initial 24-hour blood transfusion volume,Lac,and white blood cell(WBC)in poor prognosis group were higher than those in good prognosis group,while the admission GCS score and hemoglobin(Hb)level were lower than those in good prognosis group,and the differences were statistically significant(P<0.05).Pearson correlation analysis showed that the initial 24-hour blood transfusion volume was negatively correlated with admission GCS score and admission Hb level(P<0.05),and positively correlated with admission Lac level(P<0.05).The initial 24-hour blood transfusion volume,admission GCS score,admission Lac,and admission Hb levels are all independent risk factors affecting the prognosis of trauma emergency hemorrhagic shock patients(P<0.05).The initial 24-hour blood transfusion volume,admission GCS score,admission Lac,admission Hb level,and combined predicted area under the curve(AUC)were 0.722,0.872,0.881,0.798,and 0.931,respectively,with sensitivity of 68.2%,76.6%,85.7%,75.7%,and 88.8%,and specificity of 70.8%,81.0%,78.5%,81.0%,and 85.7%,respectively.Paired Z-tests showed that the combined AUC was higher than a single indicator,and both sensitivity and specificity were optimal(P<0.05).Conclusion The initial 24-hour blood transfusion volume,admission GCS score,admission Lac,and admission Hb levels are all independent risk factors for poor prognosis in trauma emergency hemorrhagic shock patients,and the combined prediction of the four has the highest efficacy value.
9.Hippo pathway-manipulating neutrophil-mimic hybrid nanoparticles for cardiac ischemic injury via modulation of local immunity and cardiac regeneration.
Qiaozi WANG ; Yanan SONG ; Jinfeng GAO ; Qiyu LI ; Jing CHEN ; Yifang XIE ; Zhengmin WANG ; Haipeng TAN ; Hongbo YANG ; Ning ZHANG ; Juying QIAN ; Zhiqing PANG ; Zheyong HUANG ; Junbo GE
Acta Pharmaceutica Sinica B 2023;13(12):4999-5015
The promise of regeneration therapy for restoration of damaged myocardium after cardiac ischemic injury relies on targeted delivery of proliferative molecules into cardiomyocytes whose healing benefits are still limited owing to severe immune microenvironment due to local high concentration of proinflammatory cytokines. Optimal therapeutic strategies are therefore in urgent need to both modulate local immunity and deliver proliferative molecules. Here, we addressed this unmet need by developing neutrophil-mimic nanoparticles NM@miR, fabricated by coating hybrid neutrophil membranes with artificial lipids onto mesoporous silica nanoparticles (MSNs) loaded with microRNA-10b. The hybrid membrane could endow nanoparticles with strong capacity to migrate into inflammatory sites and neutralize proinflammatory cytokines and increase the delivery efficiency of microRNA-10b into adult mammalian cardiomyocytes (CMs) by fusing with cell membranes and leading to the release of MSNs-miR into cytosol. Upon NM@miR administration, this nanoparticle could home to the injured myocardium, restore the local immunity, and efficiently deliver microRNA-10b to cardiomyocytes, which could reduce the activation of Hippo-YAP pathway mediated by excessive cytokines and exert the best proliferative effect of miR-10b. This combination therapy could finally improve cardiac function and mitigate ventricular remodeling. Consequently, this work offers a combination strategy of immunity modulation and proliferative molecule delivery to boost cardiac regeneration after injury.
10.TONG Xiaolin's Cold-Damp Epidemic Differentiation and Treatment Concepts and Diagnostic and Therapeutic Key Points
Chongxiang XUE ; Yingying YANG ; Jinli LUO ; Linhua ZHAO ; Xiuyang LI ;
Journal of Traditional Chinese Medicine 2023;64(22):2354-2358
Building on a clear understanding of the diagnosis and treatment patterns for cold-damp epidemics, and incorporating the insights of Academician TONG Xiaolin regarding the differentiation and treatment of such epidemics, this article have distilled the key diagnostic principles. These principles primarily entail observing color and pulse, initially distinguishing between yin and yang, with treatment focusing on dispelling cold and dampness. A comprehensive approach is adopted throughout treatment, with a combination of overall strategies and tailored treatments that evolve in line with the patient's condition. In critical and severe cases, particular attention is given to aspects such as sweating, pulse analysis, and consciousness, preventing the pathogen from penetrating deeply. Efforts are made to open and close organ networks, expelling external pathogens without fixating on resolving constipation. The dosage and efficacy of herbal remedies are adjusted based on individual circumstances and clinical symptoms. The objective is to eliminate pathogens comprehensively, while being vigilant against relapses, ultimately achieving a full recovery. This overview of the thought processes and clinical principles for diagnosing and treating cold-damp epidemics will guide precise clinical practices and provide a theoretical foundation for the involvement of traditional Chinese medicine in epidemic prevention and treatment.

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