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A novel alternative for pyrogen detection based on a transgenic cell line

Signal Transduction and Targeted Therapy 2024年第3期9卷 1221-1229页

作者： Qing He Chuan-Fei Yu Gang Wu Kai-Qin Wang Yong-Bo Ni Xiao Guo Zhi-Hao Fu Lan Wang De-Jiang Tan Hua Gao Can Wang Gang Chen Xu-Hong Chen Bo Chen Jun-Zhi Wang National institutes for food and drug control BeijingChina School of Life Science and Biopharmaceutics Shenyang Pharmaceutical UniversityShenyangChina Shanghai institute for food and drug control ShanghaiChina KeyMed Biosciences Inc. ChengduChina

Pyrogen,often as a contaminant,is a key indicator affecting the safety of almost all parenteral drugs(including biologicals,chemicals,traditional Chinese medicines and medical devices).It has become a goal to completely replace the in vivo rabbit pyrogen test by using the in vitro pyrogen test based on the promoted‘reduction,replacement and refinement’principle,which has been highly considered by regulatory agencies from different countries.We used NF-κB,a central signalling molecule mediating inflammatory responses,as a pyrogenic marker and the monocyte line THP-1 transfected with a luciferase reporter gene regulated by NF-κB as an in vitro model to detect pyrogens by measuring the intensity of a fluorescence signal.Here,we show that this test can quantitatively and sensitively detect endotoxin(lipopolysaccharide from different strains)and nonendotoxin(lipoteichoic acid,zymosan,peptidoglycan,lectin and glucan),has good stability in terms of NF-κB activity and cell phenotypes at 39 cell passages and can be applied to detect pyrogens in biologicals(group A&C meningococcal polysaccharide vaccine;basiliximab;rabies vaccine(Vero cells)for human use,freeze-dried;Japanese encephalitis vaccine(Vero cells),inactivated;insulin aspart injection;human albumin;recombinant human erythropoietin injection(CHO Cell)).The within-laboratory reproducibility of the test in three independent laboratories was 85%,80%and 80%and the interlaboratory reproducibility among laboratories was 83.3%,95.6%and 86.7%.The sensitivity(true positive rate)and specificity(true negative rate)of the test were 89.9%and 90.9%,respectively.In summary,the test provides a novel alternative for pyrogen detection.

关键词： pyrogen rabbit replace

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Overexpression of wild-type HRAS drives non-alcoholic steatohepatitis to hepatocellular carcinoma in mice

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Zoological Research 2024年第3期45卷 551-566页

作者： Chen Ling Su-Su Liu Yu-Ya Wang Gui-Tao Huo Yan-Wei Yang Nan Xu Hong Wang Yong Wu Yu-Fa Miao Rui Fu Yu-Wei Zhao Chang-Fa Fan College of Life Sciences Northwest UniversityProvincial Key Laboratory of Biotechnology of Shaanxi ProvinceXi’anShaanxi 710069China Division of Animal Model Research Institute for Laboratory Animal ResourcesNational Institutes for Food and Drug Control(NIFDC)Beijing 102629China National Center for Safety Evaluation of drugs National Institutes for Food and Drug Control(NIFDC)Beijing 100176China Division of HIV/AIDS and Sexually Transmitted Virus Vaccines Institute for Biological Products ControlNational Institutes for Food and Drug Control(NIFDC)Beijing 102629China Division of Laboratory Animal Monitoring Institute for Laboratory Animal ResourcesNational Institutes for Food and Drug Control(NIFDC)Beijing 102629China

Hepatocellular carcinoma(HCC),a prevalent solid carcinoma of significant concern,is an aggressive and often fatal disease with increasing global incidence rates and poor therapeutic outcomes.The etiology and pathological progression of non-alcoholic steatohepatitis(NASH)-related HCC is multifactorial and multistage.However,no single animal model can accurately mimic the full NASH-related HCC pathological progression,posing considerable challenges to transition and mechanistic studies.Herein,a novel conditional inducible wild-type human HRAS overexpressed mouse model(HRAS-HCC)was established,demonstrating 100%morbidity and mortality within approximately one month under normal dietary and lifestyle conditions.Advanced symptoms of HCC such as ascites,thrombus,internal hemorrhage,jaundice,and lung metastasis were successfully replicated in mice.In-depth pathological features of NASH-related HCC were demonstrated by pathological staining,biochemical analyses,and typical marker gene detections.Combined murine anti-PD-1 and sorafenib treatment effectively prolonged mouse survival,further confirming the accuracy and reliability of the model.Based on protein-protein interaction(PPI)network and RNA sequencing analyses,we speculated that overexpression of HRAS may initiate the THBS1-COL4A3 axis to induce NASH with severe fibrosis,with subsequent progression to HCC.Collectively,our study successfully duplicated natural sequential progression in a single murine model over a very short period,providing an accurate and reliable preclinical tool for therapeutic evaluations targeting the NASH to HCC continuum.

关键词： HRAS THBS1 HCC driver factor NASH Fibrosis Cirrhosis HCC Treatment evaluation

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Oroxyloside protects against dextran sulfate sodium-induced colitis by inhibiting ER stress via PPARγ activation

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Chinese Journal of Natural Medicines 2024年第4期22卷 307-317页

作者： TAO Lei DOU Renjie CHEN Xueming CAO Yu DAI Zhen HU Ziyan MA Zhi GE Xiaoming ZHANG Ling WANG Xiaoping Nanjing institute for food and drug control Nanjing 211198China State Key Laboratory of Natural Medicines School of Basic Medicine and Clinical PharmacyChina Pharmaceutical UniversityNanjing 211198China

Ulcerative colitis (UC), a prevalent form of inflammatory bowel disease (IBD), may result from immune system dysfunction, leading to the sustained overproduction of reactive oxygen species (ROS) and subsequent cellular oxidative stress damage. Recent studies have identified both peroxisome proliferator-activated receptor-γ (PPARγ) and endoplasmic reticulum (ER) stress as critical targets for the treatment of IBD. Oroxyloside (C22H20O11), derived from the root of Scutellaria baicalensis Georgi, has traditionally been used in treating inflammatory diseases. In this study, we investigated the molecular mechanisms by which oroxyloside mitigates dextran sulfate sodium (DSS)-induced colitis. We examined the effects of oroxyloside on ROS-mediated ER stress in colitis, including the protein expressions of GRP78, p-PERK, p-eIF2α, ATF4, and CHOP, which are associated with ER stress. The beneficial impact of oroxyloside was reversed by the PPARγ antagonist GW9662 (1 mg·kg^(−1), i.v.) in vivo. Furthermore, oroxyloside decreased pro-inflammatory cytokines and ROS production in both bone marrow-derived macrophages (BMDM) and the mouse macrophage cell line RAW 264.7. However, PPARγ siRNA transfection blocked the anti-inflammatory effect of oroxyloside and even abolished ROS generation and ER stress activation inhibited by oroxyloside in vitro. In conclusion, our study demonstrates that oroxyloside ameliorates DSS-induced colitis by inhibiting ER stress via PPARγ activation, suggesting that oroxyloside might be a promising effective agent for IBD.

关键词： Oroxyloside ER stress PPARγ Colitis

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Strategies for improving extraction capacity through preorganization structure:A novel 5,6-bicyclicmalonamide extractant(THPPD)

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Journal of Rare Earths 2024年第4期42卷 768-774页

作者： Miaomiao Zhang Zhen Yang Yuhua Cui Jianhui Su Heng Zhao Chengjin Xu Xiaolei Liu Guoxin Sun Xiujing Peng Yu Cui School of Chemistry and Chemical Engineering University of JinanJinan250022China institute for Smart Materials&Engineering University of JinanJinan250022China Shandong institute for food and drug control Jinan250101China

Rational design of extractant structure to improve the extraction and separation capacity of rare earth elements(REEs)is a long-standing challenge.Herein,a new strategy was proposed to improve the stability of the chelating structure formed by malonamide and REEs.The stability of the chelating structure is greatly improved by using a double-ring framework structure which makes the two carbonyl groups of malonamide unable to rotate freely and both of them point to the metal ion position.Three benzyl groups were used to construct the hydrophobicity of the extractant and the organic shell of the extracted species.Tribenzylhexahydro-pyrrolo-pyridine-dione(THPPD)was designed and synthesized.The structure reduces the energy consumed by rotating carbonyl group in the coordination with metal ions and then improves the extraction ability of extractant.The crystal structure was preorganized as expected,with the two carbonyls pointing in a favorable coordination direction to the structural complement of the metal ion.The extraction behaviors of REEs with THPPD in a nitric acid medium were studied.Compared with N,N’-dibenzyl-N,N’-dimethylmalonamide(DBDM-MA)with a chain structure,the extraction capacity of THPPD is 360 times higher than that of DBDM-MA at 5.0 mol/L sodium nitrate.Furthermore,the binding energy and Gibbs free energy were investigated by density functional theory(DFT)in conjunction with the B3LYP.The theoretical results show that THPPD has more effective interaction with Pr(NO_(3))3 than DBDM-MA.The construction of chelating groups conformation is a worthy direction to improve the coordination ability and even selectivity of extractant.

关键词： Rare earth elements Extraction separation 5,6-bicyclicdiamide extractant

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Development and Validation of an Impedimetric Sensor Modified with Molecularly Imprinted Polymer for Direct Determination of Bisphenol A in Bioanalytics

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Journal of Analysis and Testing 2024年第2期8卷 228-236页

作者： Wenjing Song Yingyi Guo Pengyu Liu Xiucong Pei School of Public Health Shenyang Medical CollegeShenyang110034China Shenyang institute for food and drug control Shenyang110021China Key Laboratory of food Pollutant Inspection and Risk Assessment in Liaoning Province Shenyang110021China

There is a strong correlation between bisphenol A(BPA)and the risks of endocrine disruption in humans.However,the mechanism behind this correlation is still unknown due to the challenge of accurately assessing BPA levels in biological matrices.Herein,we demonstrate a molecularly imprinted polymer(MIP)modified impedimetric sensor treated with bovine serum albumin(BSA)not only reduces the impact of protein interference but also maintains both specificity and sensitivity for the precise analysis of BPA.The platinum disc electrode was coated with MIP via electrochemical polymerization of o-phenylenediamine(o-PD)in the presence of BPA as a template,and free sites were then blocked with 1%BSA.Electrochemical impedance spectroscopy(EIS)was utilized for the detection of BPA.The linear range for detection was between 1.0 pg/L and 10.0 ng/L,with a detection limit(LOD)of 0.28 pg/L.The sensor presented a good selectivity over structurally similar steroid hormones and BPA metabolites.To validate the performance of the proposed sensor in the presence of possible interfering proteins,it was used to quantify BPA in real samples without any prior treatment.These findings indicate that such an impedimetric sensor can be used to measure BPA in complex matrices.

关键词： Bisphenol A Molecularly imprinted polymers Impedimetric sensor Bovine serum albumin Biological matrices

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Characterization of a pangolin SARS-CoV-2-related virus isolate that uses the human ACE2 receptor

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Science China(Life Sciences) 2024年第7期67卷 1502-1513页

作者： Luo-Yuan Xia Xue-Feng Wang Xiao-Ming Cui Yi-Ming Zhang Zhen-Fei Wang En-Tao Li Chang-Fa Fan Ke Song Yuan-Guo Li Run-Ze Ye Fang-Xu Li Dai-Yun Zhu Jie Zhang Zhuang-Zhuang Shi Ming-Zhu Zhang Liang-Jing Li Shi-Jing Shen Song Jin Ya-Wei Zhang Wei-Guang Fu Lin Zhao Wen-Hao Wang Tie-Cheng Wang You-Chun Wang Jia-Fu Jiang Yan-Ling Hu Na Jia Yu-Wei Gao Wu-Chun Cao institute of EcoHealth School of Public HealthCheeloo College of MedicineShandong UniversityJinan 250012China Changchun Veterinary Research institute Changchun 130122China State Key Laboratory of Pathogen and Biosecurity Beijing Institute of Microbiology and EpidemiologyBeijing 100071China Division of Animal Model Research Institute for Laboratory Animal ResourcesNational Institutes for Food and Drug ControlBeijing 102629China National institutes for food and drug control Beijing 102629China Life Sciences institute Guangxi Medical UniversityNanning 530020China College of Life Sciences Shandong Normal UniversityJinan 250014China Research Unit of Discovery and Tracing of Natural Focus Diseases Chinese Academy of Medical SciencesBeijing 100071China

Various SARS-CoV-2-related coronaviruses have been increasingly identified in pangolins,showing a potential threat to humans.Here we report the infectivity and pathogenicity of the SARS-CoV-2-related virus,PCoV-GX/P2V,which was isolated from a Malayan pangolin(Manis javanica).PCoV-GX/P2V could grow in human hepatoma,colorectal adenocarcinoma cells,and human primary nasal epithelial cells.It replicated more efficiently in cells expressing human angiotensin-converting enzyme 2(hACE2)as SARS-CoV-2 did.After intranasal inoculation to the hACE2-transgenic mice,PCoV-GX/P2V not only replicated in nasal turbinate and lungs,but also caused interstitial pneumonia,characterized by infiltration of mixed inflammatory cells and multifocal alveolar hemorrhage.Existing population immunity established by SARS-CoV-2 infection and vaccination may not protect people from PCoV-GX/P2V infection.These findings further verify the hACE2 utility of PCoV-GX/P2V by in vivo experiments using authentic viruses and highlight the importance for intensive surveillance to prevent possible cross-species transmission.

关键词： pangolin SARS-CoV-2-related virus infectivity pathogenicity human cell lines human ACE2 receptor hACE2-transgenic mice

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Geographical origin identification of winter jujube(Ziziphus jujuba Dongzao')by using multi-element fingerprinting with chemometrics

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Journal of Integrative Agriculture 2024年第5期23卷 1749-1762页

作者： Xiabing Kong Qiusheng Chen Min Xu Yihui Liu Xiaoming Li Lingxi Han Qiang Zhang Haoliang Wan Lu Liu Xubo Zhao Jiyun Nie College of Horticulture Qingdao Agricultural University/Laboratory of Quality&Safety Risk Assessment for Fruit(Qingdao)Ministry of Agriculture and Rural Affairs/National Technology Centre for Whole Process Quality Control of FSEN Horticultural Products(Qingdao)/Qingdao Key Lab of Modern Agriculture Quality and Safety EngineeringQingdao 266109China institute of Agricultural Product Quality Safety and NutritionTianjin Academy of Agricultural SciencesTianjin 300381China College of food Science and Engineering Northwest A&F UniversityYangling 712100China

Winter jujube(Ziziphus jujuba'Dongzao')is greatly appreciated by consumers for its excellent quality,but brand infringement frequently occurs in the market.Here,we first determined a total of 38 elements in 167 winter jujube samples from the main winter jujube producing areas of China by inductively coupled plasma mass spectrometer(ICP-MS).As a result,16 elements(Mg,K,Mn,Cu,Zn,Mo,Ba,Be,As,Se,Cd,Sb,Ce,Er,Tl,and Pb)exhibited significant differences in samples from different producing areas.Supervised linear discriminant analysis(LDA)and orthogonal projection to latent structures discriminant analysis(OPLS-DA)showed better performance in identifying the origin of samples than unsupervised principal component analysis(PCA).LDA and OPLS-DA had a mean identification accuracy of 87.84 and 94.64%in the testing set,respectively.By using the multilayer perceptron(MLP)and C5.0,the prediction accuracy of the models could reach 96.36 and 91.06%,respectively.Based on the above four chemometric methods,Cd,Tl,Mo and Se were selected as the main variables and principal markers for the origin identification of winter jujube.Overall,this study demonstrates that it is practical and precise to identify the origin of winter jujube through multi-element fingerprint analysis with chemometrics,and may also provide reference for establishing the origin traceability system of other fruits.

关键词： winter jujube multi-element fingerprint analysis chemometrics origin traceability

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Impacts of Integrated Multi-Trophic Aquaculture on Phytoplankton in Sanggou Bay

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Journal of Ocean University of China 2024年第3期23卷 835-843页

作者： HAN Jichang LIU Xing PAN Kehou LIU Jiaojiao SUN Yan JIN Guiyong LI Yun LI Yuhang College of food Science and Engineering Ningbo UniversityNingbo 315832China Key Laboratory of Mariculture(Ocean University of China Ministry of EducationQingdao 266003China Function Laboratory for Marine Fisheries Science and food Production Processes Qingdao National Labo-ratory for Marine Science and TechnologyQingdao 266100China Laboratory of Marine Organism Taxonomy and Phylogeny Institute of OceanologyChinese Academy of SciencesQingdao 266071China

Integrated multi-trophic aquaculture(IMTA)has been considered as an ecofriendly culture system providing a potential solution to environmental risks caused by intensive monoculture system.However,the impact of IMTA on phytoplankton remains unclear.In this study,the spatial and temporal variations of phytoplankton in Sanggou Bay were investigated seasonally based on 21 sampling sites covering three cultivation zones(bivalve zone,IMTA zone,and kelp zone)and one control zone(without aquatic cultivation).In total,128 phytoplankton species,with diatoms and dinoflagellates as the dominant groups,were obtained across the whole year,and the mean Shannon diversity index(H')and species richness(SR)were determined as 1.39 and 9.39,respectively.The maximum chlorophyll a(Chl-a)(6.32μg L^(-1))and plankton diversity(H'of 1.97)occurred in summer and autumn,respectively.Compared to other zones,the bivalve zone displayed significantly higher Chl-a and lower H'in majority of time.Pairwise PERMANOVA analysis indicated that the phytoplankton assemblage in the bivalve zone was significantly different with the control and kelp zones,while the IMTA zone maintained close to other three zones.Based on generalized additive models,temperature,NO_(2)^(-)-N,N/P ratio,SiO_(3)^(2-)-Si,and salinity were determined as the key factors underlying Chl-a and phytoplankton diversity.Addi-tionally,the results of redundancy analysis further indicated that the phytoplankton assemblage in the bivalve zone is positively re-lated with nutrients such as NO_(3)^(-)-N and NH_(4)^(+)-N as well as water depth,while the phytoplankton assemblages in the kelp,control,and IMTA zones are associated with NO_(2)^(-)-N,SiO_(3)^(2-)-Si,and salinity.Taken all observations into consideration together,it can be inferred that IMTA can effectively reduce Chl-a level compared to bivalve monoculture by reducing the nutrients.However,the SR,H’,and species composition of phytoplankton are primarily determined by local environment factors such as temperature,water depth,salinity a

关键词： phytoplankton diversity Sanggou Bay integrated multi-trophic aquaculture

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Higher intakes of lysine,threonine and valine are inversely associated with non-alcoholic fatty liver disease risk:a community-based case-control study in the Chinese elderly

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food Science and Human Wellness 2024年第1期13卷 191-197页

作者： Xueqi Li Wenjun Ma Ting Yang Chong Wang Wei Zhang Hui Li Ting Zhao Xiaofei Guo institute of Nutrition&Health School of Public HealthQingdao UniversityQingdao 260071China Affiliated Hospital of qingdao University Qingdao 260071China

The associations of individual amino acid with non-alcoholic fatty liver disease(NAFLD)risk remained unclear.The present study aimed to investigate the associations between the two in the Chinese elderly.Methods:A community-based health check-up program was conducted in qingdao,China.NAFLD was diagnosed by ultrasonography accompanied by epidemiological investigation.The dietary intakes of amino acids were investigated with 3-day,24-h dietary records and calculated by Nutrition Calculator software.Restricted cubic spline model was used to evaluate a nonlinear relationship between amino acid intake and NAFLD risk.Results:400 NAFLD subjects were identified,and 400 participants were randomly selected as controls and matched by gender and age(±3 years)Dose-response analysis showed that 1000 mg increment of aromatic amino acids(AAAs)was associated with reduced 16%risk of NAFLD.Dietary increments of 750 mg/d threonine,950 mg/d valine,or 1700 mg/d lysine were associated with a 20%reduction in the NAFLD risk(all P for linearity<0.05).Conclusion:The present study demonstrated that the dietary increases in milk,eggs and deep-sea fish,which are rich in the amino acids,might contribute to protecting against NAFLD in the elderly.

关键词： Amino acid Non-alcoholic fatty liver disease Senior citizen Dose-response analysis

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Anti-inflammatory mechanism of the non-volatile ingredients performance liquid chromatography-heated electron spray ionization-high resolution mass spectroscope and cell metabolomics

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Journal of Traditional Chinese Medicine 2024年第2期44卷 260-267页

作者： JING Wenguang LIN Xiaoyu LI Chu ZHAO Xiaoliang CHENG Xianlong WANG Penglong WEI Feng MA Shuangcheng institute of control of Chinese Traditional Medicine and Ethnic Medicine National Institutes for Food and Drug ControlBeijing 102629China School of Traditional Chinese Medicine Beijing University of Chinese MedicineBeijing 102488China Beijing Key Laboratory of Traditional Chinese Medicine Basic Research on Prevention and Treatment of Major Diseases Experimental Research CenterChina Academy of Chinese Medical SciencesBeijing 100700China

OBJECTIVE:To explore the anti-inflammatory components and mechanism of the non-volatile ingredients of patchouli.METHODS:High performance liquid chromatographyheated electron spray ionization-high resolution mass spectroscope(HPLC-HESI-HRMS)was used to analyze the chemical constituents of the non-volatile ingredients of patchouli.The anti-inflammatory activity of ingredients was evaluated using lipopolysaccharide(LPS)induced RAW264.7 cell inflammation model,and the antiinflammatory mechanism was investigated using multivariate statistical analysis of cell metabolomics.RESULTS:The non-volatile ingredients of patchouli were characterized by HPLC-HESI-HRMS,and 36 flavonoids and 18 other components were identified.These ingredients of patchouli not only had a good protective effect on the LPS-induced inflammation model of RAW264.7 cells,but also regulated the expression levels of arginine,L-leucine,cholesterol,fructose and sorbitol by down-regulating arginine metabolism,aminoacyl-tRNA biosynthesis,polyol/sorbitol pathway,so as to reduce inflammation and reduce cell damage.CONCLUSION:The non-volatile ingredients of patchouli had good anti-inflammatory effect and exerted its curative effect by regulating endogenous metabolic pathway to reduce inflammatory response.

关键词： Pogostemon non-volatile ingredients anti-inflammatory cell metabolomics

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