1.The mechanism of extract of ginkgo biloba inducing mitochondrial autophagy in breast cancer cells MCF-7
Jianqiang SHAO ; Peng WANG ; Jie BAI ; Huixin LI ; Zunyi WANG ; Zhihong XU
Journal of International Oncology 2024;51(2):65-72
Objective:To investigate the mechanism of extract of ginkgo biloba (EGB) on mitochondrial autophagy in breast cancer cells MCF-7.Methods:Breast cancer MCF-7 cells were divided into four groups. EGB with mass concentrations of 40, 80, 120 mg/L was used to incubate breast cancer MCF-7 cells for 24 h or 48 h, as a low concentration group of EGB, a medium concentration group of EGB, and a high concentration group of EGB. Breast cancer MCF-7 cells without intervention were taken as control group. Cell proliferation was measured using MTT assay; Flow cytometry was used to detect cell apoptosis; Immunofluorescence assay was used to determine the contents of prostacyclin (P62), microtubule-associated protein light chain 3Ⅱ (LC3Ⅱ), and caspase-3; The levels of multidrug resistance-associated protein 1 (MRP1), multidrug resistance gene 1 (MDR1) and breast cancer resistance protein (BCRP) were identified by PCR; Western blotting was used to detect the expression of extracellular signal-regulated kinase (ERK), mitogen-activated protein kinase (MAPK), p-ERK, and p-MAPK proteins in cells.Results:The results of MTT assay for cell proliferation showed that cell proliferation at 24 h in control group, EGB low, medium and high concentration groups were 0.95±0.14, 0.65±0.09, 0.51±0.07, 0.37±0.04, respectively, with a statistically significant difference ( F=43.13, P<0.001), cell proliferation at 48 h were 1.32±0.19, 0.54±0.08, 0.32±0.05, 0.15±0.02, respectively, with a statistically significant difference ( F=141.30, P<0.001). Compared with 24 h, cell proliferation was decreased in EGB low, medium and high concentration groups at 48 h (all P<0.05). Pairwise comparison showed that EGB treatment significantly decreased MCF-7 cell viability and cell proliferation was decreased in turn at 24 and 48 h in control group, low, medium, high EGB groups (all P<0.05). Flow cytometry analysis revealed that the apoptosis rates of MCF-7 cells in control group, EGB low, medium and high concentration groups were 2.12%±0.23%, 9.28%±0.45%, 15.17%±1.28% and 22.21%±2.32%, respectively, with a statistically significant difference ( F=128.80, P<0.001). Pairwise comparison showed that the apoptosis rate of control group, EGB low, medium and high concentration groups were increased in turn (all P<0.05). The results of immunofluorescence assay showed that the protein relative expression levels of P62 protein in MCF-7 cells of control group, EGB low, medium and high concentration groups were 3.34±0.52, 2.85±0.47, 2.02±0.18 and 1.08±0.21, respectively, with a statistically significant difference ( F=41.55, P<0.001). LC3Ⅱ protein relative expression levels were 0.24±0.05, 1.02±0.14, 1.47±0.26, 1.95±0.21, respectively, with a statistically significant difference ( F=94.82, P<0.001). The relative expression levels of caspase-3 protein were 0.25±0.03, 0.68±0.21, 1.12±0.17 and 1.65±0.23, respectively, with a statistically significant difference ( F=68.09, P<0.001). Pairwise comparison showed that LC3Ⅱ and caspase-3 protein expression levels were increased in turn in control group, EGB low, medium and high concentration groups, while P62 protein expression levels were decreased in turn (all P<0.05). The PCR experiment results showed that the MRP1 mRNA level of MCF-7 cells in control group, EGB low, medium and high concentration groups were 1.06±0.14, 0.83±0.18, 0.71±0.11, 0.52±0.08, respectively, with a statistically significant difference ( F=17.41, P<0.001). The mRNA levels of MDR1 were 1.14±0.17, 0.75±0.13, 0.60±0.09, 0.48±0.06, respectively, with a statistically significant difference ( F=34.40, P<0.001). BCRP mRNA levels were 1.09±0.11, 0.88±0.13, 0.69±0.07, 0.57±0.05, respectively, with a statistically significant difference ( F=34.13, P<0.001). Pairwise comparison showed that the levels of MRP1, MDR1 and BCRP mRNA were decreased in turn in control group, EGB low, medium and high concentration groups (all P<0.05). The results of Western blotting showed that the expression of ERK in MCF-7 cells in control group, EGB low, medium and high concentration groups were 2.54±0.38, 1.89±0.25, 1.55±0.21, 1.12±0.16, respectively, with a statistically significant difference ( F=31.18, P<0.001). MAPK expression were 2.47±0.34, 1.96±0.29, 1.63±0.27, 1.20±0.24, respectively, with a statistically significant difference ( F=20.90, P<0.001). p-ERK expression were 2.03±0.29, 1.74±0.21, 1.45±0.11, 1.18±0.24, respectively, with a statistically significant difference ( F=16.31, P<0.001). p-MAPK expression were 2.26±0.47, 1.90±0.41, 1.61±0.33, 1.35±0.16, respectively, with a statistically significant difference ( F=7.01, P=0.002). Pairwise comparison showed that the expressions of ERK, MAPK, p-ERK and p-MAPK in control group, EGB low, medium and high concentration groups were decreased in turn (all P<0.05) . Conclusion:EGB can inhibit the proliferation of breast cancer MCF-7 cells, promote the apoptosis of MCF-7 cells, decrease the expression of P62 protein, increase the expression of LC3Ⅱ and caspase-3 protein, induce mitochondrial autophagy.
2.Correlation analysis of serum Chemerin with disease activity and Th17/Treg in patients with rheumatoid arthritis
Wenjing XU ; Dongmei GAO ; Huixin LI ; Li WANG ; Shengquan TONG
Tianjin Medical Journal 2024;52(2):193-196
Objective To explore the correlation of serum Chemerin level with disease activity and the ratio of T helper 17/regulatory T cells(Th17/Treg)in patients with rheumatoid arthritis(RA).Methods A total of 180 patients with RA who were admitted to our hospital were regarded as the observation group.According to the DAS28 score,the observation group was divided into the high activity group(60 cases),the moderate activity group(60 cases)and the low activity group(60 cases).Another 180 healthy people who underwent physical examination in our hospital during the same period were regarded as the control group.Enzyme-linked immunosorbent assay(ELISA)was used to detect serum levels of Chemerin,interleukin-9(IL-9),interleukin-10(IL-10)and interleukin-17(IL-17).Flow cytometry was used to detect the Th17/Treg ratio.Spearman correlation analysis was applied to analyze the correlation between serum Chemerin level and DAS28 score.Pearson correlation analysis was used to analyze the correlation between serum Chemerin level and Th17,Treg cell percentage and Th17/Treg ratio.Results The results of this study showed that the serum level of Chemerin was higher in the observation group than that in the control group(P<0.05).The serum Chemerin level was positively correlated with DAS28 score(P<0.05).Serum Chemerin levels and DAS28 scores decreased in turn in the high,moderate and low activity groups(P<0.05).The percentage of Th17 cells and the ratio of Th17/Treg were higher in the observation group than those in the control group,and the percentage of Treg cells was lower in the observation group than that in the control group(P<0.05).The level of IL-10 was lower in the observation group than that in the control group,while levels of IL-17 and IL-9 were higher in the observation group than those in the control group(P<0.05).The results of Pearson correlation analysis showed that serum Chemerin level was positively correlated with the percentage of Th17 cells and the ratio of Th17/Treg,and negatively correlated with the percentage of Treg cells(P<0.05).Conclusion Serum Chemerin level is elevated in patients with RA,which is closely related to disease activity and Th17/Treg ratio.
3.Application of near infrared responsive hydrogels in tissue engineering
Jing XU ; Huixin LYU ; Xin BAO ; Yi ZHANG ; Yihan WANG ; Yanmin ZHOU
Chinese Journal of Tissue Engineering Research 2024;28(3):486-492
BACKGROUND:Near infrared responsive hydrogels,have a variety of excellent properties such as high spatial and temporal precision,remote tunability,and safety and non-invasiveness,providing a new direction of exploration for the development of tissue engineering. OBJECTIVE:To summarize the application progress of near infrared responsive hydrogels in the field of tissue engineering in recent years. METHODS:The literature search was performed on PubMed and CNKI databases.The keywords were"near infrared responsive hydrogels,tissue engineering,bone defect,bone repair,bone regeneration,wound healing,wound dressing,angiogenesis"in Chinese and English.The search time limit was from May 2006 to October 2022 and extended for some classical literature.The abstracts and contents of the retrieved literature were analyzed,and the relevant literature was obtained according to inclusion and exclusion criteria.Finally,97 articles were included for review. RESULTS AND CONCLUSION:(1)Near infrared responsive materials are involved in tissue repair by controlling infection and reducing inflammation,promoting angiogenesis,osteoblast differentiation and new bone formation.(2)Near infrared responsive hydrogel can be prepared by constructing a thermosensitive hydrogel with a photothermal effect or by using a photochemical reaction.(3)Near infrared responsive hydrogels as wound dressings perform various functions such as rapid hemostasis,tissue adhesion through polymerization of polymer monomers,antibacterial and anti-inflammatory effects,and promotion of angiopoiesis and epithelial regeneration through the local photothermal effect of photothermal nanomaterials during soft tissue healing and regeneration.(4)Near infrared responsive hydrogels function during bone reconstruction and repair by promoting osteogenic differentiation of mesenchymal stem cells,stimulating the expression of heat shock proteins,and increasing angiogenesis.(5)Near infrared responsive hydrogels present a combination of multiple therapeutic strategies with significant synergistic therapeutic functions and are also being progressively developed for application in other tissue reconstruction and disease treatment scenarios.
4.Research and exploration on the construction path of multi-campus smart hospital
Yanting ZHAO ; Yake LU ; Linpeng YANG ; Xinpei YUE ; Min ZHAO ; Shuoguo WANG ; Huixin HAN ; Xinkui LIU
Modern Hospital 2024;24(3):420-423
The construction of smart hospital is an important part of modern hospital management system,and it is also the key way to build the new system of high-quality hospital development.In terms of building smart hospitals,multi-campus hos-pitals face more difficulties and challenges than single campus hospitals,such as the lack of top-level design,the difficulty of in-tegrated management,the uneven development of hospitals and the widespread phenomenon of information islands.This study summarizes and analyzes the difficulties encountered in the construction and application of smart hospitals in multi-hospital areas.Guided by problems,it puts forward countermeasures and suggestions for the construction of refined and high-quality smart hospi-tals in multi-campus hospitals,including strengthening overall and forward-looking awareness,integrating management according to hospital conditions,characteristic development under demand guidance,establishing a data integration center for smart hospi-tals,scientific planning of talent reserve and discipline layout,etc.
5.Foundation of ceRNA networks and functional validation of AFAP1-AS1 in lung adenocarcinoma
Huixin WANG ; Qian LI ; Xiaowen HOU ; Xinzhu SHI ; Xu FENG
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2024;31(04):576-584
Objective A competing endogenous RNA (ceRNA) regulatory network associated with long non-coding RNA (lncRNA) specific for lung adenocarcinoma (LUAD) was constructed based on bioinformatics methods, and the functional mechanism of actinfilament-associated protein 1-antisense RNA1 (AFAP1-AS1) in LUAD was analyzed, in order to provide a new direction for the study of LUAD therapeutic targets. Methods The gene chip of LUAD was downloaded from the Gene Expression Omnibus (GEO), and lncRNA and mRNA with differential expression between LUAD and normal tissues were screened using GEO2R online software, and their target genes were predicted by online databases to construct ceRNA networks and perform enrichment analysis. In cell experiments, AFAP1-AS1 was genetically knocked down and siRNA was constructed and transfected into LUAD cells A549 by cell transfection. CCK8, transwell, scratch assay and flow cytometry were used to detect the ability of cells to proliferate, invade, migrate and apoptosis. Results A total of 6 differentially expressed lncRNA and 494 differentially expressed mRNA were identified in the microarray of LUAD. The ceRNA network involved a total of 6 lncRNA, 22 miRNA, and 55 mRNA. Enrichment analysis revealed that mRNA was associated with cancer-related pathways. In cell assays, knockdown of AFAP1-AS1 inhibited cell proliferation, invasion, and migration, and AFAP1-AS1 promoted apoptosis. Conclusion In this study, we construct a lncRNA-mediated ceRNA network, which may help to further investigate the mechanism of action of LUAD. In addition, through cellular experiments, AFAP1-AS1 is found to have potential as a therapeutic target for LUAD.
6.The Effect of Yiqi Huoxue Formula (益气活血方) in the Treatment of Chronic Pulmonary Heart Disease and Heart Failure with Qi Deficiency and Blood Stasis Pattern of 58 Cases: A Randomized Controlled Trial
Weitao CHEN ; Yanpeng ZENG ; Lianhua SU ; Hong ZHANG ; Huixin WANG ; Juanjuan ZHANG
Journal of Traditional Chinese Medicine 2024;65(10):1019-1025
ObjectiveTo explore the effectiveness and safety of Yiqi Huoxue Formula (益气活血方, YHF) in the adjuvant treatment of chronic pulmonary heart disease (CPHD) and heart failure (HF)with qi deficiency and blood stasis pattern. MethodsOne hundred and twenty patients with CPHD and HF with qi deficiency and blood stasis pattern were allocated randomly into treatment group and control group, with 60 case in each group. The control group was given conventional basic western medicine, while the treatment group was given oral administration of YHF granules in addition, one dose per day. The treatment course for both groups was 8 weeks. The TCM symptom scores, Minnesota Life Quality Scale (MLHF-Q) scores, echocardiographic indicators including right ventricular end-diastolic diameter (RVEDD), left ventricular end-diastolic diameter (LVEDD), left atrial end-diastolic diameter (LAEDD) and pulmonary artery mean pressure (PAMP), six-minute walking distance (6MWD), and plasma N-terminal pro-B-type natriuretic peptide (NT-ProBNP) level were compared between the groups. The effectiveness regarding cardiac function and TCM syndromes were compared between the two groups after treatment, and the occurrence of adverse events was observed. ResultsWith two drop-outs both in the treatment group and control group, and 58 cases in each group were included in the outcome analysis. The total effective rate regarding cardiac function and TCM syndromes in the treatment group were 91.38% (53/58) and 96.55% (56/58), respectively, significantly higher than the corresponding 70.69% (41/58) and 48.27% (28/58) in the control group (P<0.05). After treatment, the TCM symptom scores and RVEDD level were significantly reduced in the treatment group, and MLHF-Q score, plasma NT-ProBNP level and PAMP level decreased significantly, while 6MWD increased in both groups (P<0.01). Compared to those in the control group, the TCM symptom scores, MLHF-Q score, plasma NT-ProBNP level and PAMP level significantly decreased, while 6MWD increased in the treatment group (P<0.01). There were no obvious abnormalities in the blood, urine, stool routine and liver and kidney function indicators in both groups. One adverse reaction each occurred in both groups, and there was no statistically significant difference in the incidence rates(P>0.05). ConclusionYHF combined with conventional western medicine can significantly improve the clinical efficacy, improve the clinical symptoms and cardiac function, increase the quality of life and exercise tolerance, and is relatively safe.
7.Effects of Different Pellet Feed Hardness on Growth and Reproduction,Feed Utilization Rate,and Environmental Dust in Laboratory Mice
Dong WU ; Rui SHI ; Peishan LUO ; Ling'en LI ; Xijing SHENG ; Mengyang WANG ; Lu NI ; Sujuan WANG ; Huixin YANG ; Jing ZHAO
Laboratory Animal and Comparative Medicine 2024;44(3):313-320
Objective To study the effects of different pellet feed hardness on the growth and reproduction,feed utilization rate,and environmental dust in laboratory mice.Methods One hundred of fifty 50 3-week-old SPF-grade C57BL/6JGpt and 150 ICR laboratory mice were randomly divided into three groups,with an equal number of males and females.They were fed diets with different hardness of 18.62 kg,23.15 kg,and 27.89 kg.Body weight,feed utilization rate,and dust levels in cages were recorded and calculated for mice aged 3-10 weeks.Forty-five 6-week-old male mice and ninety 4-week-old female mice from each strain were randomly divided into three groups and fed pellet feeds with three different hardness levels.After 2 weeks of adaptation to the same hardness feed,the mice were paired at a 1:2 male-to-female ratio and monitored for reproductive data for 3 months.Results At the age of 4 weeks,the body weight of male C57BL/6JGpt mice in 23.15 kg group was significantly higher than that in the 18.62 kg and 27.89 kg groups(P<0.01),and the body weight of females in the 18.62 kg group was significantly higher than that in the 27.89 kg group(P<0.05).There was no significant difference in body weight among ICR mice aged 3-10 weeks across different feed hardness groups(P>0.05).For both strains,feed utilization rate for males was higher than that for females across different feed hardness groups at all weeks of age(P<0.01).Compared to the 27.89 kg group,both the 18.62 kg and 23.15 kg groups showed a significant increase in the 50-mesh dust levels in cages for both strains aged 4-8 weeks(except for 7-week-old C57BL/6JGpt mice)(P<0.05).For both C57BL/6JGpt and ICR mice,there was no significant difference in basic reproductive performance such as interval between the first litter and the monthly production index among the three feed hardness groups during the experimental period(P>0.05).However,the monthly production index of C57BL/6JGpt mice first increased and then decreased with the increase of feed hardness,while that of ICR mice increased with increasing feed hardness,though these differences were not statistically significant(P>0.05).Conclusion Different strains and genders had different tolerance to feed hardness.C57BL/6JGpt mice are more adapted to lower hardness feeds,while ICR mice are better suited to slightly higher hardness feeds.
8.A pilot study on clinical application of three-dimensional morphological completion of lesioned mandibles assisted by generative adversarial networks
Ye LIANG ; Qian WANG ; Yiyi ZHANG ; Jingjing HUAN ; Jie CHEN ; Huixin WANG ; Zhuo QIU ; Peixuan LIU ; Wenjie REN ; Yujie MA ; Canhua JIANG ; Jiada LI
Chinese Journal of Stomatology 2024;59(12):1213-1220
Objective:To explore the clinical application pathway of the CT generative adversarial networks (CTGANs) algorithm in mandibular reconstruction surgery, aiming to provide a valuable reference for this procedure.Methods:A clinical exploratory study was conducted, 27 patients who visited the Department of Oral and Maxillofacial Surgery, Xiangya Hospital of Central South University between January 2022 and January 2024 and required mandibular reconstruction were selected. The cohort included 16 males and 11 females, with the age of (46.6±11.5) years; among them, 7 cases involved mandibular defects crossing the midline. The CTGANs generator produced 100 images, and the mean squared error (MSE) was calculated for differences between any two generated images. Preoperative cone-beam CT data from 5 patients were used to construct a labeled test database, divided into groups: normal maxilla, normal mandible, diseased mandible, and noise (each group containing 70 cross-sectional images). The CTGANs discriminator was used to evaluate the loss values for each group, and one-way ANOVA and intergroup comparisons were performed. Using the self-developed KuYe multioutcome-option-network generation system (KMG) software, the three-dimensional (3D) completion area of the mandible under cone-beam CT was defined for the 27 patients. The CTGANs algorithm was applied to obtain a reference model for the mandible. Virtual surgery was then performed, utilizing the fibular segment to reconstruct the mandible and design the surgical expectation model. The second-generation combined bone-cutting and prebent reconstruction plate positioning method was used to design and 3D print surgical guides, which were subsequently applied in mandibular reconstruction surgery for the 27 patients. Postoperative cone-beam CT was used to compare the morphology of the reconstructed mandible with the surgical expectation model and the mandibular reference model to assess the three-dimensional deviation.Results:The MSE for the CTGANs generator was 2 411.9±833.6 (95% CI: 2 388.7-2 435.1). No significant difference in loss values was found between the normal mandible and diseased mandible groups ( P>0.05), while both groups demonstrated significantly lower loss values than the maxilla and noise groups ( P<0.001). All 27 patients successfully obtained mandibular reference models and surgical expectation models. In total, 14 162 negative deviation points and 15 346 positive deviation points were observed when comparing the reconstructed mandible morphology with the surgical expectation model, with mean deviations of -1.32 mm (95% CI:-1.33- -1.31 mm) and 1.90 mm (95% CI: 1.04-1.06 mm), respectively. Conclusions:The CTGANs algorithm is capable of generating diverse mandibular reference models that reflect the natural anatomical characteristics of the mandible and closely match individual patient morphology, thereby facilitating the design of surgical expectation models. This method shows promise for application in patients with mandibular defects crossing the midline.
9.DIP evaluation index system
Zunzun LIU ; Xinkui LIU ; Yanting ZHAO ; Linpeng YANG ; Yuzhu ZHANG ; Huixin HAN ; Shuoguo WANG
Modern Hospital 2024;24(5):703-706,710
Objective This paper aims to construct a scientific and standardized index system for evaluating the applica-tion effect of Diagnosis-Intervention Packet(DIP)in medical institutions.It seeks to determine the weights of indicators at all lev-els to provide a basis for evaluating the effectiveness of DIP used in various regions as well as a reference for improving DIP poli-cy.Methods This paper used literature analysis to develop an indicator pool preliminarily.The Delphi method was used to con-duct a questionnaire survey with 22 experts to establish the index system.It also used the Analytic Hierarchy Process(AHP)to determine the weight of each index.Results A total of two rounds of expert consultation questionnaires were conducted.The questionnaire exhibited high expert positive coefficients by 91.00%and 100.00%and higher authority coefficients of both 0.91 for two rounds.The coordination coefficients of the two rounds of expert consultation met the standard,and the expert opinions reached a consistence.Additionally,the questionnaire established comprised three primary indicators(efficiency of medical in-surance fund,medical service management,patient benefit,and satisfaction),9 secondary indicators,and 38 tertiary indicators.The AHP was used to determine the weight of indicators at all levels.Among the primary indicators,patient benefit and satisfac-tion had the highest weight(0.446 9).Among the secondary indicators,patients'medical cost burden had the highest weight(0.287 6).Among the tertiary indicators,the inpatient satisfaction had the highest weight(0.1592).The Cronbach's α coeffi-cients of the two rounds were>0.7.Conclusion This paper utilized Delphi method and AHP to establish the evaluation system for assessing the implementation effectiveness of DIP.The approach is highly scientific and authoritative.It can be used to evalu-ate the application effect of DIP and provide an effective tool for medical insurance and hospital managers at all levels to assess DIP payment policies.
10.Comparison between sepsis-induced coagulopathy and sepsis-associated coagulopathy criteria in identifying sepsis-associated disseminated intravascular coagulation
Zhao HUIXIN ; Dong YIMING ; Wang SIJIA ; Shen JIAYUAN ; Song ZHENJU ; Xue MINGMING ; Shao MIAN
World Journal of Emergency Medicine 2024;15(3):190-196
BACKGROUND:Disseminated intravascular coagulation(DIC)is associated with increased mortality in sepsis patients.In this study,we aimed to assess the clinical ability of sepsis-induced coagulopathy(SIC)and sepsis-associated coagulopathy(SAC)criteria in identifying overt-DIC and pre-DIC status in sepsis patients. METHODS:Data from 419 sepsis patients were retrospectively collected from July 2018 to December 2022.The performances of the SIC and SAC were assessed to identify overt-DIC on days 1,3,7,or 14.The SIC status or SIC score on day 1,the SAC status or SAC score on day 1,and the sum of the SIC or SAC scores on days 1 and 3 were compared in terms of their ability to identify pre-DIC.The SIC or SAC status on day 1 was evaluated as a pre-DIC indicator for anticoagulant initiation. RESULTS:On day 1,the incidences of coagulopathy according to overt-DIC,SIC and SAC criteria were 11.7%,22.0%and 31.5%,respectively.The specificity of SIC for identifying overt-DIC was significantly higher than that of the SAC criteria from day 1 to day 14(P<0.05).On day 1,the SIC score with a cut-off value>3 had a significantly higher sensitivity(72.00%)and area under the curve(AUC)(0.69)in identifying pre-DIC than did the SIC or SAC status(sensitivity:SIC status 44.00%,SAC status 52.00%;AUC:SIC status 0.62,SAC status 0.61).The sum of the SIC scores on days 1 and 3 had a higher AUC value for identifying the pre-DIC state than that of SAC(0.79 vs.0.69,P<0.001).Favorable effects of anticoagulant therapy were observed in SIC(adjusted hazard ratio[HR]=0.216,95%confidence interval[95%CI]:0.060-0.783,P=0.018)and SAC(adjusted HR=0.146,95%CI:0.041-0.513,P=0.003). CONCLUSION:The SIC and SAC seem to be valuable for predicting overt-DIC.The sum of SIC scores on days 1 and 3 has the potential to help identify pre-DIC.

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