1.2,3,5,4′-tetrahydroxyldiphenylethylene-2-O-glucoside Attenuates Cerebral Ischemia-reperfusion Injury via PINK1/LETM1 Signaling Pathway
Hongyu ZENG ; Kaimei TAN ; Feng QIU ; Yun XIANG ; Ziyang ZHOU ; Dahua WU ; Chang LEI ; Hongqing ZHAO ; Yuhong WANG ; Xiuli ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):145-154
ObjectiveTo investigate the mechanism by which 2,3,5,4'-tetrahydroxyldiphenylethylene-2-O-glucoside (THSG) mitigates cerebral ischemia/reperfusion (CI/R) injury by regulating mitochondrial calcium overload and promoting mitophagy. MethodsSixty male SD rats were randomized into sham, model, SAS (40 mg·kg-1), and low-, medium- and high-dose (10, 20, 40 mg·kg-1, respectively) THSG groups, with 10 rats in each group. The middle cerebral artery occlusion/reperfusion (MCAO/R) model was established by the modified Longa suture method. An oxygen-glucose deprivation/reoxygenation (OGD/R) model was constructed in PC12 cells. Neurological deficits were assessed via Zea Longa scoring, and cerebral infarct volume was measured by 2,3,5-triphenyltetrazolium chloride (TTC) staining. Structural and functional changes of cortical neurons in MCAO/R rats were assessed by hematoxylin-eosin and Nissl staining. PC12 cell viability was detected by cell counting kit-8 (CCK-8) assay, and mitochondrial calcium levels were quantified by Rhod-2 AM. Immunofluorescence was used to detect co-localization of PTEN-induced kinase 1 (PINK1) and leucine zipper/EF-hand-containing transmembrane protein 1 (LETM1) in neurons. Transmission electron microscopy (TEM) was employed to observe mitochondrial morphology in neurons. Western blot was employed to analyze the expression of translocase of outer mitochondrial membrane 20 (TOMM20), autophagy-associated protein p62, microtubule-associated protein light chain 3 (LC3), cysteinyl aspartate-specific proteinase-9 (Caspase-9), B-cell lymphoma 2-associated protein X (Bax), and cytochrome C (Cyt C). ResultsCompared with the sham group, the model group exhibited increased infarct volume (P<0.01) and neurological deficit scores (P<0.01), neuronal structure was disrupted with reduced Nissl bodies. (P<0.01), mitochondrial swelling/fragmentation, decreased PINK1/LETM1 co-localization (P<0.01), upregulated protein levels of LC3Ⅱ/LC3Ⅰ, TOMM20, Caspase-9, Bax, and Cyt C (P<0.01), downregulated protein level of p62 (P<0.05), weakened PC12 viability (P<0.01), and elevated mitochondrial calcium level (P<0.01). Compared with the model group, THSG and SAS groups showed reduced infarct volumes (P<0.05,P<0.01) and neurological deficit scores (P<0.05,P<0.01), mitigated mitochondrial damage, and increased PINK1/LETM1 co-localization (P<0.01). Medium/high-dose THSG and SAS alleviated the neurological damage, increased Nissl bodies (P<0.05,P<0.01), downregulated the protein levels of p62, TOMM20, Caspase-9, Bax, and Cyt C (P<0.05,P<0.01), and elevated the LC3Ⅱ/LC3Ⅰ level (P<0.05,P<0.01). High-dose THSG enhanced PC12 cell viability (P<0.01), increased PINK1/LETM1 co-localization (P<0.01), and reduced mitochondrial calcium (P<0.01). ConclusionTHSG may exert the neuroprotective effect on CI/R injury by activating the PINK1-LETM1 signaling pathway, reducing the mitochondrial calcium overload, and promoting mitophagy.
2.2,3,5,4′-tetrahydroxyldiphenylethylene-2-O-glucoside Attenuates Cerebral Ischemia-reperfusion Injury via PINK1/LETM1 Signaling Pathway
Hongyu ZENG ; Kaimei TAN ; Feng QIU ; Yun XIANG ; Ziyang ZHOU ; Dahua WU ; Chang LEI ; Hongqing ZHAO ; Yuhong WANG ; Xiuli ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):145-154
ObjectiveTo investigate the mechanism by which 2,3,5,4'-tetrahydroxyldiphenylethylene-2-O-glucoside (THSG) mitigates cerebral ischemia/reperfusion (CI/R) injury by regulating mitochondrial calcium overload and promoting mitophagy. MethodsSixty male SD rats were randomized into sham, model, SAS (40 mg·kg-1), and low-, medium- and high-dose (10, 20, 40 mg·kg-1, respectively) THSG groups, with 10 rats in each group. The middle cerebral artery occlusion/reperfusion (MCAO/R) model was established by the modified Longa suture method. An oxygen-glucose deprivation/reoxygenation (OGD/R) model was constructed in PC12 cells. Neurological deficits were assessed via Zea Longa scoring, and cerebral infarct volume was measured by 2,3,5-triphenyltetrazolium chloride (TTC) staining. Structural and functional changes of cortical neurons in MCAO/R rats were assessed by hematoxylin-eosin and Nissl staining. PC12 cell viability was detected by cell counting kit-8 (CCK-8) assay, and mitochondrial calcium levels were quantified by Rhod-2 AM. Immunofluorescence was used to detect co-localization of PTEN-induced kinase 1 (PINK1) and leucine zipper/EF-hand-containing transmembrane protein 1 (LETM1) in neurons. Transmission electron microscopy (TEM) was employed to observe mitochondrial morphology in neurons. Western blot was employed to analyze the expression of translocase of outer mitochondrial membrane 20 (TOMM20), autophagy-associated protein p62, microtubule-associated protein light chain 3 (LC3), cysteinyl aspartate-specific proteinase-9 (Caspase-9), B-cell lymphoma 2-associated protein X (Bax), and cytochrome C (Cyt C). ResultsCompared with the sham group, the model group exhibited increased infarct volume (P<0.01) and neurological deficit scores (P<0.01), neuronal structure was disrupted with reduced Nissl bodies. (P<0.01), mitochondrial swelling/fragmentation, decreased PINK1/LETM1 co-localization (P<0.01), upregulated protein levels of LC3Ⅱ/LC3Ⅰ, TOMM20, Caspase-9, Bax, and Cyt C (P<0.01), downregulated protein level of p62 (P<0.05), weakened PC12 viability (P<0.01), and elevated mitochondrial calcium level (P<0.01). Compared with the model group, THSG and SAS groups showed reduced infarct volumes (P<0.05,P<0.01) and neurological deficit scores (P<0.05,P<0.01), mitigated mitochondrial damage, and increased PINK1/LETM1 co-localization (P<0.01). Medium/high-dose THSG and SAS alleviated the neurological damage, increased Nissl bodies (P<0.05,P<0.01), downregulated the protein levels of p62, TOMM20, Caspase-9, Bax, and Cyt C (P<0.05,P<0.01), and elevated the LC3Ⅱ/LC3Ⅰ level (P<0.05,P<0.01). High-dose THSG enhanced PC12 cell viability (P<0.01), increased PINK1/LETM1 co-localization (P<0.01), and reduced mitochondrial calcium (P<0.01). ConclusionTHSG may exert the neuroprotective effect on CI/R injury by activating the PINK1-LETM1 signaling pathway, reducing the mitochondrial calcium overload, and promoting mitophagy.
3.Surveillance and analysis of microbial contamination in commercially available ready-to-eat foods in Songjiang District, Shanghai from 2017 to 2022
Xiang QIU ; Yuxing ZHU ; Xinyi GU ; Fengsong SHENG ; Yuanqiang JIANG
Shanghai Journal of Preventive Medicine 2025;37(3):255-259
ObjectiveTo investigate the microbial contamination status of commercially available ready-to-eat (RTE) foods in Songjiang District of Shanghai, to verify the potential food safety hazards, so as to provide a scientific basis for the local regulation of RTE foods and the prevention of foodborne diseases. MethodsA total of 882 food samples from 9 categories of RTE foods were collected in Shanghai from 2017 to 2022. Salmonella, Listeria monocytogenes, Vibrio parahaemolyticus, Staphyloccus aureus and Bacillus cereus were detected according to the standard operating procedures in the Manual for China National Food Contamination and Harmful Factors Risk Monitoring, and which were determined with reference to relevant standards. ResultsA total of 77 samples were found to carry foodborne pathogens, with a total detection rate of 8.73%. Among the 5 foodborne pathogens, Bacillus cereus had the highest detection rate (8.35%), followed by Vibrio parahaemolyticus (5.05%), and the difference of detection rates among the 5 foodborne pathogens was statistically significant (χ2=112.307, P<0.001). Among the 9 categories of RTE food products, the detection rate of foodborne pathogens in RTE fermented/non-fermented soybean products was the highest (27.50%), followed by cold noodles (17.33%) and raw edible aquatic products (9.50%), and the difference in the detection rates of foodborne pathogens among different RTE food categories was statistically significant (χ2=45.909, P<0.001). The detection rate of foodborne pathogens in the third quarter of the year was 10.99%, significantly higher than that in other quarters (χ2=10.329, P=0.016). The detection rate of foodborne pathogens in samples from supermarkets was highest (15.08%), followed by snack bars (11.58%), with a statistically significant difference (χ2=14.108, P=0.015). ConclusionFoodborne pathogens contamination is found in commercially available RTE foods in Songjiang District of Shanghai, especially in RTE fermented/non-fermented soybean products, RTE cold noodles and raw edible aquatic products from supermarkets, snack bars and farm products markets during summer and autumn. Therefore, it is crucial to strengthen supervision and management to avoid the foodborne diseases.
4.Functional near infrared imaging of college students on speech fluency tasks in traumatized
ZHENG Minxiao, ZHU Dongmei, WEI Qiang, XIANG Nian, QIU Min, XIAO Qiang, LI Xiaonan, ZHANG Yan
Chinese Journal of School Health 2024;45(3):375-378
Objective:
The study aims to explore the neural mechanism of cognitive differences in college students with posttraumatic stress disorder under verbal fluency task based on functional near infrared spectroscopy (fNIRS), so as to provide neuroimaging support for the evaluation, diagnosis and treatment of posttraumatic stress disorder(PTSD).
Methods:
Posttaumatic Stress Disorder Checklist Combat(PCL-C) was used to screen the subjects, including 21 students in PTSD group (PCL-C≥38) and 30 students in control group from September to Novenber in 2020. A 53 channel near infrared spectroscopy device was used to collect cerebral blood oxygen signals under the verbal fluency task, and correlation analysis, Mann Whitney U test and independent sample t test were performed on the results.
Results:
The difference in the total average score of PCL-C Scale between PTSD group and the control group(46.38±6.96,25.57±6.09) was statistically significant ( t=11.33, P <0.05). Correlation analysis showed that Avg-HbO in left dorsolateral prefrontal lobe was negatively correlated with PCL-C Score ( r=-0.37, P <0.05). Mann Whitney U test showed that in the left dorsolateral prefrontal lobe (Ch6), the Avg-HbO change in PTSD group [0.19(-0.09, 0.86)mmol/(L〖KG*7〗·mm)] was significantly lower than the control group [0.79( 0.37 , 1.47)mmol/(L ·mm)] ( Z=2.16, P <0.05), which was statistically significant.
Conclusions
The degree of PTSD was negatively correlated with the index of oxygenated hemoglobin in the left dorsolateral prefrontal lobe, and the oxygenated hemoglobin content in the PTSD group was lower than that in the normal group. In the future, fNIRS may be used to collect blood oxygen signals from the left dorsolateral prefrontal lobe in cognitive tasks to provide imaging evidence for the identification of PTSD.
5.Pathological Characteristics and Classification of Unstable Coronary Atheroscle-rotic Plaques
Yun-Hong XING ; Yang LI ; Wen-Zheng WANG ; Liang-Liang WANG ; Le-Le SUN ; Qiu-Xiang DU ; Jie CAO ; Guang-Long HE ; Jun-Hong SUN
Journal of Forensic Medicine 2024;40(1):59-63
Important forensic diagnostic indicators of sudden death in coronary atherosclerotic heart dis-ease,such as acute or chronic myocardial ischemic changes,sometimes make it difficult to locate the ischemic site due to the short death process,the lack of tissue reaction time.In some cases,the de-ceased died of sudden death on the first-episode,resulting in difficulty for medical examiners to make an accurate diagnosis.However,clinical studies on coronary instability plaque revealed the key role of coronary spasm and thrombosis caused by their lesions in sudden coronary death process.This paper mainly summarizes the pathological characteristics of unstable coronary plaque based on clinical medi-cal research,including plaque rupture,plaque erosion and calcified nodules,as well as the influencing factors leading to plaque instability,and briefly describes the research progress and technique of the atherosclerotic plaques,in order to improve the study on the mechanism of sudden coronary death and improve the accuracy of the forensic diagnosis of sudden coronary death by diagnosing different patho-logic states of coronary atherosclerotic plaques.
6.Relationship among physical activity,mild depressive symptoms and frontal alpha power asymmetry in college students
Xiang WANG ; Xiaojing ZHOU ; Shali QIU ; Yuheng ZANG ; Peng WANG ; Jing WANG ; Jinlei ZHAO ; Xin XIN ; Qun ZHAO ; Suowang YIN ; Xing WANG
Chinese Mental Health Journal 2024;38(2):180-185
Objective:To investigate the correlation among physical activity,mild depressive symptoms and frontal alpha power asymmetry in college students.Methods:Seventy college students with mild depressive symp-toms who conformed to the standard of the Self-Rating Scale for Depression(SDS)of 53-62 and 70 normal col-lege students were recruited.The frontal alpha power was measured under quiet and closed-eye state,and the total physical activity(PA)was assessed with the International Physical Activity Questionnaire.Results:The college students with mild depressive symptoms had lower Total PA scores,right frontal alpha power and frontal alpha a-symmetry(FAA)than the normal controls(P<0.001).In college students with mild depressive symptoms,the to-tal PA scores(r=-0.29,P<0.05)and FAA(r=-0.41,P<0.001)were negatively correlated with SDS scores,and the total PA scores were positively correlated with FAA(r=0.34,P<0.01).Conclusion:The college students with mild depressive symptoms may have reduced physical activity and asymmetric right lateralization of frontal alpha power.There is a correlation among depressive symptoms,physical activity and frontal alpha power a-symmetry in college students with mild depressive symptoms.
7.Advances in therapeutic drug monitoring methods based on liquid chromatography-tandem mass spectrometry
Ziying LI ; Jie XIE ; Ziyu QU ; You JIANG ; Di ZHANG ; Songlin YU ; Xiaoli MA ; Ling QIU ; Xinhua DAI ; Xiang FANG ; Xiaoping YU
Chinese Journal of Laboratory Medicine 2024;47(3):332-340
Liquid chromatography-tandem mass spectrometry (LC-MS/MS) technology has the characteristics of high specificity and high throughput, making it rapidly applied and developed in the field of clinical testing. Its application in the monitoring of therapeutic drugs can effectively improve the quantitative accuracy and sensitivity, and formulate a personalized and optimal dosing plan for patients. However, this technology still faces some challenges, and automation, quality control, and quantitative traceability will be the future development direction.
8.Comparison of Wild and Cultivated Codonopsis pilosula Based onTraditional Quality Evaluation
Xiaoyan LAN ; Chunfang TIAN ; Zhilai ZHAN ; Li ZHOU ; Xiang LI ; Zidong QIU ; Tiegui NAN ; Qili YUAN ; Xiaoliang LIN ; Congkui TIAN ; Meilan CHEN ; Liping KANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(14):156-164
ObjectiveTo conduct a systematic comparative study on wild and cultivated Codonopsis pilosula(CP) from three aspects, including characters, microscopy, and contents of primary and secondary metabolites. MethodWild and cultivated CP samples were collected, their characters were measured using vernier caliper, tape measure and balance, the paraffin sections were stained with safranin-fixed green dyeing, and their microstructure were observed under the optical microscope. The content of alcohol-soluble extracts in wild and cultivated CP was determined according to the method for determination of extract under CP in the 2020 edition of Chinese Pharmacopoeia, the starch content was determined by anthrone colorimetry, the content of total polysaccharides was determined by kit method, Fiber analyzer was used to determine the content of fiber components, and ultra performance liquid chromatography(UPLC) was used to determine the content of monosaccharides, disaccharides and some secondary metabolites. Multivariate statistical analysis methods such as principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were employed to screen key differential components between wild and cultivated CP on the basis of variable importance in the projection(VIP) value>1 and P<0.05. ResultIn terms of morphological characteristics, the "lion's head-like" shape, longitudinal wrinkles, and circumferential wrinkles below the root cap of wild CP were more pronounced in wild CP compared to the cultivated ones. Regarding transverse sectional features, wild CP had more fissures on the outer side of the cortex and a larger duramen. Under microscopic examination, wild CP had more stone cells, a larger proportion of xylem, and the presence of cork cells arranged in rings in the xylem, while cultivated CP has a larger proportion of phloem, smaller vessel diameters, and a more loosely arranged vascular system. In terms of primary metabolites, the contents of 45% ethanol-soluble extract and total polysaccharides in cultivated CP were significantly higher than those in the wild ones(P<0.05), the contents of lignin, hemicellulose, cellulose, fructose and glucose in wild CP were significantly higher than those in the cultivated ones(P<0.05), while sucrose content in the cultivated CP was significantly higher than that in the wild ones(P<0.05). Concerning secondary metabolites, the contents of tryptophan and tangshenoside Ⅰ in cultivated CP were significantly higher than those in the wild ones(P<0.05), whereas the contents of lobetyolinin, lobetyol and atractylenolide Ⅲ in wild CP were significantly higher than those in the cultivated ones(P<0.05). ConclusionThere are significant differences between wild and cultivated CP in terms of morphological characteristics, microscopic features and chemical composition. Glucose, fructose, sucrose, tangshenoside Ⅰ, tryptophan and cellulose components are the key differential components between wild and cultivated CP. Wild CP contains more polyacetylenes and fructose, whereas cultivated CP has higher levels of tangshenoside Ⅰ and sucrose, with noticeably lower cellulose content. These distinctions may be related to their growth conditions, growth years and cultivation techniques. Based on the results of this study, it is recommended to increase polyacetylenes and the content ratio of fructose to sucrose as an indicators to characterize different production methods of CP, in order to guide the high-quality production of CP.
9.Comparison of Wild and Cultivated Paeoniae Radix Rubra Based on Traditional Quality Evaluation
Chunfang TIAN ; Qiannan HU ; Zhilai ZHAN ; Xiaoyan LAN ; Xiang LI ; Li ZHOU ; Tiegui NAN ; Zidong QIU ; Liping KANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(14):165-174
ObjectiveTo compare wild and cultivated Paeoniae Radix Rubra(PRR) in three aspects, including character, microscope, determination of primary and secondary metabolites. MethodSeventeen batches of wild and nine batches of cultivated PRR were collected,their character data were measured by vernier caliper and scales, and their paraffin sections were made by safranin-fixed green dyeing for the observation of microscopic features. The content of ethanol-soluble extracts and total tannin from wild and cultivated PRR was determined by the method of general principle 2201 and 2202 in the 2020 edition of Chinese Pharmacopoeia, the content of polysaccharides was determined by phenol-sulfuric acid method. Anthrone colorimetry was used to determine the content of starch, and Van Soest method of washing fiber was used to determine the content of fiber. The contents of fructose, glucose and sucrose in wild and cultivated PRR were determined by ultra-high performance liquid chromatography evaporative light scattering detection(UPLC-ELSD), and the secondary metabolites(gallic acid, methyl gallate, catechin, oxypaeoniflorin, albiflorin, paeoniflorin, ellagic acid, 1,3,4,6-tetragalloylglucose, galloylpaeoniflorin, 1,2,3,4,6-O-pentagalloylglucose, naringenin, benzoylpaeoniflorin and benzoylalbiflorin) were determined by UPLC. Principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were used to analyze the data of wild and cultivated PRR, the contribution of different factors to the difference was determined according to the variable importance in the projection(VIP) value>1 and P<0.05. ResultIn term of characters, wild PRR showed the traditional characteristic of Zaopi Fencha, its outer skin was loose and easy to fall off, its surface had longitudinal furrow and wrinkle, but the outer skin of cultivated PRR was not easy to fall off, and its surface was relatively smooth. The radial texture of xylem of wild PRR cross-section was more obvious, showing radial striations, vacuoles and more cracks, while the radial texture of xylem of cultivated PRR cross-section was not obvious, dense and some had cracks. Microscopically, the number of radial vessels arranged in the xylem of wild PRR was more than that of cultivated PRR, the number of calcium oxalate clusters in the phloem and xylem of wild PRR was more than that of cultivated PRR, while the number of starch grains was significantly higher in cultivated PRR. In terms of the content of primary chemical constituents, the contents of polysaccharides and starch of cultivated PRR were significantly higher than those of wild PRR(P<0.05), while the contents of cellulose, lignin, fructose and glucose of wild PRR were significantly higher than those of cultivated PRR(P<0.05). The results of determination of 13 secondary metabolites showed that the contents of paeoniflorin, methyl gallate, catechin and oxypaeoniflorin in wild PRR were significantly higher than those in cultivated PRR(P<0.05), while the contents of albiflorin, gallic acid, ellagic acid, naringenin, benzoylpaeoniflorin and benzoylalbiflorin were significantly lower than those of cultivated PRR(P<0.05). A total of 10 variables contributing to the differentiation between wild and cultivated PRR were screened, including albiflorin, cellulose, benzoylpaeoniflorin, oxypaeoniflorin, naringenin, ellagic acid, starch, lignin, paeoniflorin and total tannins. ConclusionThere are significant differences between wild and cultivated PRR in characters, microscopic characteristics, contents of primary and secondary metabolites. It is suggested that the content ratio of paeoniflorin and albiflorin, the contents of oxypaeoniflorin and cellulose can be used as indicators to characterize production methods of PRR so as to improve the quality standard of PRR. This study can provide reference for the improvement of quality standard of PRR and the guidance of high quality production of PRR.
10.The Role of Mechanical Sensitive Ion Channel Piezo in Digestive System Diseases
Si-Qi WANG ; Xiang-Yun YAN ; Yan-Qiu LI ; Fang-Li LUO ; Jun-Peng YAO ; Pei-Tao MA ; Yu-Jun HOU ; Hai-Yan QIN ; Yun-Zhou SHI ; Ying LI
Progress in Biochemistry and Biophysics 2024;51(8):1883-1894
The Piezo protein is a non-selective mechanosensitive cation channel that exhibits sensitivity to mechanical stimuli such as pressure and shear stress. It converts mechanical signals into bioelectric activity within cells, thus triggering specific biological responses. In the digestive system, Piezo protein plays a crucial role in maintaining normal physiological activities, including digestion, absorption, metabolic regulation, and immune modulation. However, dysregulation in Piezo protein expression may lead to the occurrence of several pathological conditions, including visceral hypersensitivity, impairment of intestinal mucosal barrier function, and immune inflammation.Therefore, conducting a comprehensive review of the physiological functions and pathological roles of Piezo protein in the digestive system is of paramount importance. In this review, we systematically summarize the structural and dynamic characteristics of Piezo protein, its expression patterns, and physiological functions in the digestive system. We particularly focus on elucidating the mechanisms of action of Piezo protein in digestive system tumor diseases, inflammatory diseases, fibrotic diseases, and functional disorders. Through the integration of the latest research findings, we have observed that Piezo protein plays a crucial role in the pathogenesis of various digestive system diseases. There exist intricate interactions between Piezo protein and multiple phenotypes of digestive system tumors such as proliferation, apoptosis, and metastasis. In inflammatory diseases, Piezo protein promotes intestinal immune responses and pancreatic trypsinogen activation, contributing to the development of ulcerative colitis, Crohn’s disease, and pancreatitis. Additionally, Piezo1, through pathways involving co-action with the TRPV4 ion channel, facilitates neutrophil recruitment and suppresses HIF-1α ubiquitination, thereby mediating organ fibrosis in organs like the liver and pancreas. Moreover, Piezo protein regulation by gut microbiota or factors like age and gender can result in increased or decreased visceral sensitivity, and alterations in intestinal mucosal barrier structure and permeability, which are closely associated with functional disorders like irritable bowel sydrome (IBS) and functional consitipaction (FC). A thorough exploration of Piezo protein as a potential therapeutic target in digestive system diseases can provide a scientific basis and theoretical support for future clinical diagnosis and treatment strategies.


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