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.Brassica juncea WRKY12 mediates bolting and flowering by interacting with the SOC1 and FUL promoters.
Yifang HUANG ; Yue DONG ; Yue YU ; Dakun LIU ; Qinlin DENG ; Yuanda WANG ; Dayong WEI ; Zhimin WANG ; Qinglin TANG
Chinese Journal of Biotechnology 2025;41(7):2818-2828
Flowering and bolting are important agronomic traits in cruciferous crops such as Brassica juncea. Timely flowering can ensure the crop organ yield and quality, as well as seed propagation. The WRKY family plays an important role in regulating plant bolting and flowering, while the function and mechanism of WRKY12 in B. juncea remain unknown. To explore its function and mechanism in bolting and flowering of B. juncea, we cloned and characterized the BjuWRKY12 gene in B. juncea and found that its expression levels were significantly higher in flowers and inflorescences than in leaves. BjuWRKY12 belonged to the Ⅱc subfamily of the WRKY family, and subcellular localization indicated that the protein was located in the nucleus. Ectopic overexpression of BjuWRKY12 in transgenic lines promoted bolting and flowering, leading to significant increases in the expression levels of flowering integrators SOC1 and FUL. Furthermore, yeast one-hybrid and dual luciferase reporter system assays confirmed that BjuWRKY12 directly bound to the promoters of BjuSOC1 and BjuFUL, undergoing protein-DNA interactions. This discovery gives new insights into the regulation network and molecular mechanisms of BjuWRKY12, laying a theoretical foundation for the breeding of high-yield and high-quality varieties of B. juncea.
Mustard Plant/metabolism*
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Flowers/growth & development*
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Plant Proteins/physiology*
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Promoter Regions, Genetic/genetics*
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Gene Expression Regulation, Plant
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Plants, Genetically Modified/genetics*
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Transcription Factors/metabolism*
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MADS Domain Proteins/metabolism*
3.Comparison of the predictive value of venous thromboembolism assessment tools in medical inpatients
Yifang HOU ; Xiaomei DENG ; Jun DUAN ; Ping ZHANG
Chinese Journal of Practical Nursing 2024;40(10):758-764
Objective:A comparison was made between the predictive efficacy of the Padua Score and the simplified Assessment Scheme Recommended by Chinese experts (hereinafter referred to as the Simplified Method) for the risk assessment of venous thromboembolism (VTE) in medical inpatients, aiming to provide a reference for the clinical selection of appropriate risk assessment tools.Methods:A retrospective cohort study was conducted, selecting 42 257 internal medicine inpatients discharged from Peking University Shenzhen Hospital between May 1, 2021, and April 30, 2022, using a convenience sampling method. Data collected included general information upon admission, VTE-related information, occurrences of VTE during hospitalization, and results from the two assessment tools. The predictive efficacy of the tools was evaluated by plotting ROC curves and calculating AUC, sensitivity, specificity, positive predictive value, negative predictive value, and predictive accuracy.Results:Among 42 257 patients, there were 21 065 male and 21 192 female participants, aged (55.04 ± 15.17) years old. The incidence rate of VTE among medical inpatients was 2.24% (948/42 257). The AUC for Padua Score and the Simplified Method in medical patients were 0.735 (95% CI 0.717-0.753) and 0.582 (95% CI 0.561-0.602), respectively. Sensitivities were 49.4% and 18.2%, specificities were 89.6% and 98.1%, positive predictive values were 9.9% and 17.7%, negative predictive values were 98.7% and 98.1%, and predictive accuracy were 88.7% and 96.3%, respectively. The departments with the highest incidence rates of VTE during hospitalization were rehabilitation medicine, emergency, neurology, geriatrics, and respiratory medicine. Within these departments, the AUC values for the Padua Score and the Simplified Method were as follows: 0.864 and 0.612, 0.782 and 0.653, 0.792 and 0.664, 0.850 and 0.551, 0.867 and 0.664, respectively. Conclusions:The Padua Score demonstrated better predictive efficacy compared to the Simplified Method. However, the Simplified Method had more accessible assessment criteria and could serve as an initial VTE risk screening tool in emergency situations or when complete data are not available.
4.Contrast analysis of chemical constituents between single decoction and mixed decoction of Inulae Flos - Haematitum medicinal pair with different proportion
Yifang DENG ; Weimei CHEN ; Qiaohua CHEN ; Kaidong WANG ; Liuying QIN ; Zan YANG ; Liaoyuan LIU
International Journal of Traditional Chinese Medicine 2023;45(3):323-329
Objective:To compare the differences of chemical components between single decoction and mixed decoction with different compatibility ratio of Inulae Flos- Haematitum medicinal pair. Methods:UPLC method was used to determine the contents of 5-caffeoylquinic acid, chlorogenic acid, 4-dicaffeoylquinic acid, caffeic acid, isoquercitrin, isochlorogenic acid B, 1,5- dicaffeoyl quinic acid, isochlorogenic acid C and the fingerprints of the single decoctions and mixed decoctions of Inulae Flos- Haematitum medicinal pair in four groups of proportions. The "peak area/sample weight" value of each common peak in the fingerprints was calculated, and the SPSS 26.0 was used for independent-sample t-test analysis. Results:There are significant differences in the "peak area/weight" values of peak 1, peak 2, peak 4, peak 6 , peak 9, peak 10, peak 12, peak 13, peak 15 between mixed decoction and single decoction of Inulae Flos - Haematitum medicinal pair with different compatibility ratios ( P<0.05), with statistical significance; when the compatibility ratio of Inulae Flos- Haematitum medicinal pair was 3:1, the difference of fingerprints and index components content between single decoction and combined decoction was the largest. Except for peak 7 and peak 14, the difference of "peak area/sample weight" value of other characteristic peaks was statistically significant ( P<0.05), and the content difference of 8 index components was statistically significant ( P<0.05). Conclusion:There are differences in the chemical components of Inulae Flos - Haematitum medicinal pair for single decoction and mixed decoction.
5.Retrospective analysis of first venous thromboembolism risk assessment in hospitalized surgical patients
Xiaomei DENG ; Jun DUAN ; Yonghuan HU ; Chengzhang HUANG ; Yifang HOU
Chinese Journal of Practical Nursing 2022;38(21):1651-1656
Objective:To investigate the assessment and occurrence of first-occured venous thromboembolism(VTE) among hospitalized patients.Methods:The clinical data of 6 532 surgical patients in Shenzhen Hospital, Peking University who were admitted from May 1, 2021 to June 30, 2021 were collected and analyzed retrospectively. The demographic data, Caprini score at admission and the incidence of VTE during hospitalization were analyzed by two independent sample t test and chi square test. Results:The Caprini score at admission of 6 532 patients was 1.81 ± 1.71. The number of cases in high, medium and low risks was 363 (5.6%), 1 189 (18.2%), 4 980 (76.2%), respectively. There was significant difference in VTE risk assessment scores and grades in different gender ( t=5.31, χ 2=48.31), length of stay ( F=195.21, χ 2=548.52) and hypertension ( t=17.07, χ 2=280.89), diabetes ( t=12.14, χ 2=51.18), smoking ( F=31.71, χ 2=53.23) and drinking ( F=18.78, χ 2=30.07) ( P<0.05). Forty-four(0.7%) patients got hospital-acquired VTE totally, among which, 24 cases (6.6%) were in high-risk, 14 cases (1.2%) were in medium-risk and 6 cases (0.1%) were in low-risk. What′s more, the top five VTE risky departments based on the assessment were not completely consistent with the top five departments with the highest incidence of VTE. Conclusions:The hospitalized patients are at high risk of VTE. The risk factors of diabetes, hypertension, smoking, drinking and other related factors should be included in the evaluation model. Meanwhile, the VTE risk assessment of in-patients should be emphasized and prophylactic treatments should be taken to reduce the incidence of VTE.
6.Development and psychometric analysis of the negative emotion screening scale for inpatients
Xiaomei DENG ; Jingping ZHANG ; Yifang HOU ; Ming WU ; Xiulan DENG ; Lingyu HOU
Chinese Journal of Behavioral Medicine and Brain Science 2021;30(8):745-750
Objective:To develop a negative emotion screening scale for inpatients(NESSI) and test its validity and reliability.Methods:Based on our previous studies and the theory model of psychological stress, the original item pool was established through literature review, expert interviews and patient consultation.The first version of NESSI was constructed by Delphi method, then initially tested in 421 inpatients followed by the project analysis and reliability test. After those above, the formal scale was developed and tested in 318 inpatients followed by confirmatory factor analysis and reliability test.Finally, 7-item generalized anxiety disorder scale (GAD-7), 9-item patient health questionnaire (PHQ-9), anger state expression scale (SAS) and simplified Chinese version of fear of disease progression scale(FoP-Q-SF) were used to test the criterion validity.Results:After exploratory factor analysis, 17 items were retained in the final scale, which can be categorized into four dimensions: fear of illness, depression, somatization and anger, which could explain 63.49% of the total variation.Confirmatory factor analysis showed that the fitting degree of each factor model was good and met the requirements of reference value (χ 2/ df=2.949, RMR=0.044, CFI=0.929, NFI=0.897, IFI=0.930, TLI=0.915, PGFI=0.655, RMSEA=0.078). The Cronbach's α coefficient of the total scale was 0.925, and the Cronbach's α coefficient of the four factors ranged from 0.762 to 0.898.The criterion validity showed that there was a significant positive correlation between the scale and the four criterion scales ( r= 0.574-0.805, all P<0.01). Conclusion:The NESSI scale has good reliability and validity, and can be used as a psychological problem screening tool among non-psychiatric inpatients.
7.Identification and Study on Processing End-point of Raw and Processed Products of Morus alba Based on UPLC Fingerprint and Chromaticity
Mengting HUANG ; Ling PAN ; Lihong DENG ; Mingyan XIE ; Yongfu MA ; Mei WEI ; Xueren CHENG ; Jie XU
China Pharmacy 2021;32(1):56-63
OBJECTIVE:To provide reference for the identification and proces sing end-point determination of raw Morus alba and its processed products (honey-processed M. alba ). METHODS :UPLC method was adopted. The determination was performed on Waters BEH Shield RP C 18 column with mobile phase consisted of acetonitrile- 0.1% phosphoric acid solution (gradient elution ) at the flow rate of 0.30 mL/min. The column temperature was set at 30 ℃. The program wavelengths were set at 280 nm(0-4 min) and 320 nm(4-35 min). Similarity Evaluation System for Chromatogram Fingerprint of TCM (2012 edition)was used to establish UPLC fingerprint and carry out similarity evaluation of 13 batches of M. alba and honey-processed M. alba . The chromatographic peaks were identified with reference substance fingerprint. The colorimetric value (L,a,b) of 13 batches of M. alba and honey-processed M. alba powder were determined ,and average total colorimetric value (E)was calculated. OPLS-DA and cluster analysis were adopted to analyze the differences in fingerprints and colorimetric values of M. alba before and after processing. At the same time ,the dynamic change rule of fingerprint and colorimetric value of honey-processed M. alba at different processing time points were analyzed to determine the processing end-point. RESULTS :There were obvious differences in fingerprints before and after processing ,and the similarity of 13 batches of M. alba and honey-processed M. alba were all higher than 0.9. Totally 21 common peaks were calibrated for M. alba ,and 23 common peaks for honey-processed M. alba ;peak 1 and peak 2 were newly produced compounds of honey-processed M. alba . Peak 2,peak 7,peak 14 and peak 19 were identified as 5-hydroxymethylfurfural, mulberry glucoside A ,oxidized resveratrol ,mulberry flavonoids G. Results of OPLS-DA showed that the peak area-sample quantity ratio of peak 1,peak 2,peak 18,peak 20 and the chromaticity values (L,a,b)were the most important factors affecting the difference of raw and processed products of M. alba . When the E ranged 75.84-80.88 as the processing end-point of honey-processed M. alba ,the processing time was determined as 22-34 min. CONCLUSIONS : The established UPLC fingerprint and colorimetric value determination method can be used to identify the raw and processed products of M. alba as well as determine the processing end-point of honey-processed M. alba .
8.Study on UHPLC Characteristic Chromatograms of Euodia rutaecarpa ,Processed E. rutaecarpa Decoction Piece , Water Decoction and Formula Granules
Lihong DENG ; Zhipeng ZHANG ; Qingqing TIAN ; Xueren CHENG ; Yun LU ; Liaoyuan LIU
China Pharmacy 2020;31(14):1739-1745
OBJECTIVE:To e stablish UPLC characteristic chrom atograms of Euodia rutaecarpa ,processed E. rutaecarpa decoction piece ,water decoction and formula granules ,and to compare its relationship and difference. METHODS :UPLC method was used. The determination was performed on YMC Triart C 18 column with mobile phase consisted of acetonitrile- 0.1% phosphoric acid water solution (gradient elution )at the flow rate of 0.3 mL/min. The detection wavelength was set at 254 nm,and column temperature was 30 ℃. The sample size was 1 μL. Using limonin as reference,the characteristic chromatograms of E. rutaecarpa , processd E. rutaecarpa decoction piece ,water decoction and formula granules (each 10 batches,totally 60 batches)were drawn. The similarity was evaluated with TCM Chromatographic Fingerprint Similarity Evaluation System (2012 edition),and to determine the common characteristic peak. The differences of ratio of common characteristic peak area were evalucoted according to variance analysis. Meanwhile ,the cluster analysis and principal component analysis (PCA) were performed to research the differences of E. rutaecarpa ,processed E. rutaecarpa decoction piece ,water decoction and formula granules by using SPSS 20.0 software. RESULTS :Totally 16 and 17 common peaks were respectively confirmed in characteristic chromatograms of E. rutaecarpa samples and processed E. rutaecarpa samples(decoction piece ,water decoction and formula granules ). No. 8,9,11, 17 peaks were identified as limonin ,evodiamine,rutaecarpine and glycyrrhizic acid. Compared with decoction piece ,the similarities of characteristic peak between water decoction and formula granules were lower than 0.55,while those between water decoction and formula granule were higher than 0.95. Cluster analysis and PCA results showed that E. rutaecarpa decoction piece and processed E. rutaecarpa decoction piece could be clustered into one category ;E. rutaecarpa water decoction and formula granules could be clustered into one category ;processed E. rutaecarpa water decoction and formula granules could be clustered into one category. CONCLUSIONS :Compared with E. rutaecarpa ,processed E. rutaecarpa additionally contain glycyrrhizic acid ; main che mical components of decoction piece ,water decoction and formula granules are basically the same ,but the contents of the components between decoction piece to water decoction and formula granules are different.
9.Effects of Chaihu Shugan powder on hepatic lipid metabolism and AMPK/SIRT1 pathway in rats with non-alcoholic fatty liver disease
Yupei ZHANG ; Yuanjun DENG ; Chaofeng HU ; Li HAN ; Qinhe YANG ; Yinji LIANG ; Yilin KONG ; Yifang HE ; Ling JIN ; Yuanyuan LI ; Guanlong WANG ; Shaobing CHENG ; Guifang TU
Chinese Journal of Pathophysiology 2016;32(2):307-313
[ ABSTRACT] AIM:To investigate the effects of Chaihu Shugan powder ( CSP) on lipid metabolism and the pro-teins involved in adenosine 5’-monophosphate-activated protein kinase (AMPK)/sirtuin 1 (SIRT1) pathway in the liver tissues of the rats with non-alcoholic fatty liver disease (NAFLD).METHODS: Sprague-Dawley rats were randomly di-vided into normal control ( NC) group, with HFD ( HFD) group and CSP group.The NAFLD models were established by feeding with HFD for 16 weeks in the rats.The rats in CSP group were intragastrically administered with CSP extracts (9.6 g· kg-1 · d-1 ) , and blood and liver samples were collected 16 weeks later.Serum and liver levels of total cholesterol ( TC) and triglyceride ( TG) , and serum levels of alanine aminotransferase ( ALT) and aspartate aminotransferase ( AST) were measured using an automatic biochemical analyzer.The histological changes of liver tissues were observed with HE staining, while the lipid deposition was observed with Oil Red O staining.The ultrastructural changes of the liver tissues were observed under transmission electron microscope.Moreover, the protein levels of AMPK, phosphorylated AMPK (pAMPK), SIRT1 and uncoupling protein 2 (UCP2) in the liver were detected by Western blot.RESULTS:The results of HE staining, Oil Red O staining and electron microscopy demonstrated that NAFLD rat model was successfully estab-lished.Compared with NC group, the serum and liver levels of TC and TG, and serum level of AST in model group were markedly elevated ( P<0.01) .Moreover, the protein levels of pAMPK and SIRT1 in HFD group were markedly reduced (P<0.01), whereas UCP2 level was elevated (P<0.01).Furthermore, liver levels of TC and TG, and serum level of AST in GSP group were markedly reduced as compared with HFD group ( P<0.05 ) .The protein levels of pAMPK and SIRT1 were elevated ( P<0.05 ) , whereas the UCP2 level was reduced as compared with HFD group ( P<0.01 ) .The protein level of AMPK between the 3 groups had no significant difference.CONCLUSION: CSP attenuates hepatic lipid disorder and hepatic lipid deposition in NAFLD rats induced by feeding with HFD for 16 weeks, which is associated with the activation of AMPK/SIRT1 pathway.
10.Clinical research of neutrophil parameters Neut-X and Neut-Y in patients with blood stream infection
Guifang CHEN ; Wenjun DENG ; Yifang ZHAI ; Yanrong ZHENG ; Fengzhi ZHENG
International Journal of Laboratory Medicine 2016;37(23):3283-3284,3287
Objective To investigate the clinical significance of neutrophil parameters Neut‐X and Neut‐Y in the diagnosis of blood stream infection(BSI) .Methods The data in106 patients with blood stream infection and 51 cases in the healthy control group were retrospectively analyzed .The results of WBC ,neutrophil percentage ,Neut‐X ,Neut‐Y and procaclitonin (PCT ) were compared .Results Neut‐X and Neut‐Y in the BSI group were 1 393 .5 ± 33 .4 and 416 .2 ± 30 .0 respectively ,which were signifi‐cantly higher than 1 371 .9 ± 32 .7 and 391 .7 ± 23 .7 in bacterial culture negative group and 1 347 .2 ± 26 .2 and 371 .9 ± 21 .5 in the healthy control group ,the differences were statistically significant (P<0 .05) .Neut‐X and Neut‐Y had good correlation with PCT . Neut‐X and Neut‐Y levels in the Gram‐negative bacterial group were higher than those in the Gram‐positive bacterial group .Conclu‐sion Neut‐X and Neut‐Y parameters in the patients with BSI are significantly increased and can be used as the assistant diagnosis parameters of blood stream infection .

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