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1.
The objective of this study was to explore the immunomodulatory effects of betulinic acid (BA) extracted from the bark of white birch on mice. Female mice were orally administered BA for 14 days in doses of 0, 0.25, 0.5, and 1 mg/kg body weight. We found that BA significantly enhanced the thymus and spleen indices, and stimulated lymphocyte proliferation induced by Concanavalin A and lipopolysaccharide as shown by MTT assay. Flow cytometry revealed that BA increased the percentage of CD4+ cells in thymus as well as the percentage of CD19+ and the ratios of CD4+/CD8+ in spleen. BA increased the number of plaque-forming cell and macrophage phagocytic activity as indicated by a neutral red dye uptake assay, and the peritoneal macrophages levels of TNF-α were also increased. In contrast, serum levels of IgG and IgM and serum concentrations of IL-2 and IL-6 were significantly decreased in BA-treated mice compared to the control as assayed by haemagglutination tests and ELISA, respectively. Taken together, these results suggest that BA enhances mouse cellular immunity, humoral immunity, and activity of macrophages. Thus, BA is a potential immune stimulator and may strengthen the immune response of its host.  相似文献   

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Alterations of genes are known to be critical for the induction of tumorigenesis, but the mechanism of ovarian carcinogenesis is little understood and remains to be elucidated. In this study, we investigated the roles of brca1, brca2 and p53 genes in the development of ovarian cancer using conditional knockout mice generated by a Cre-loxP recombinant system. Following the application of recombinant adenovirus expressing Cre in vitro, the proliferation of ovarian surface epithelium (OSE) was increased. For instance, a significant increase in cell growth was observed in OSE cells in vitro by conditional knockout isolated from the mice bearing concurrent floxed copies of brca1 and brca2/p53. However, the proliferative effect of the ovarian cells was not observed in concurrent brca1/brca2 or p53 knockout mice in vivo, indicating that we could not observe the direct evidence of the involvement of brca1, brca2, and p53 in ovarian carcinogenesis. Since morphological changes including tumor formation were not observed in mice bearing floxed copies of concurrent brca1/brca2 or p53, the inactivation of brca1/2 or p53 is not sufficient for the induction of tumor formation. Taken together, these results suggest that the deficiency of these genes may not be involved directly in the mechanism of ovarian carcinogenesis.  相似文献   

4.
To study the canine immune system we generated a mouse model engrafted with canine lymphocytes using NOD SCID IL2R common gamma chain ?/? (NSG) mice as recipients (Ca-PBL-SCID). Engraftment of canine peripheral blood lymphocytes (PBLs) was determined post-injection with 107 peripheral blood mononuclear cells (PBMCs) into irradiated NSG mice using flow cytometry and fluorescently labeled antibodies specific to canine helper T cells (CD45+ CD4+), cytotoxic lymphocytes (CD45+ CD8+), regulatory T cells (CD45+ CD4+ Foxp3+), and B cells (CD45+ Ig+ CD21lo). Canine CD45+ lymphocytes were detectable as early as day 1 in the peritoneal cavity, and beginning at 9 days in the blood, bone marrow, and spleen. CD4+ T cells, of which Foxp-3+ CD25hi cells constituted a minor percentage, were the predominant lymphocyte population at 9 days post engraftment contrasting with increasing proportions of CD8+ CTL's and Ig+ B cells beginning at 16 days. Canine immunoglobulin was initially detected in the serum of Ca-PBL-SCID mice at 9 days post-engraftment and peaked in concentration at day 36. From day 28 to 52 post-engraftment 30% of the Ca-PBL-SCID mice became markedly anemic and thrombocytopenic, yet gross and histopathologic examination of bone marrow, kidneys, spleen, liver, and intestine revealed no obvious lesions. Blood smear evaluation revealed agglutination of mature red blood cells, reticulocytes and a regenerative anemia. These findings demonstrate that NSG mice are capable of engraftment of canine PBLs yet develop graft versus host disease similar to Hu-PBL-SCID mice.  相似文献   

5.
The thymus is a primary lymphoid organ where the primary T cell repertoire is generated. Thymus atrophy is induced by various conditions, including infectious diseases, glucocorticoid treatment, and poor breeding management. Cattle with thymus atrophy tend to exhibit weak calf syndrome, a condition in which approximately half of neonates die shortly after birth. Calves with thymus atrophy that survive the first month typically contract chronic inflammatory diseases. In this study, we analyzed the populations of the peripheral blood mononuclear cells and thymocytes in calves with thymus atrophy. In addition, we evaluated polarization of master gene and cytokine mRNA expression in peripheral blood CD4+ cells in the calves. The population of CD4+CD8+ cells in thymus of the calves with thymus atrophy was lower than that of control calves. IL10 mRNA expression in peripheral blood CD4+ cells of calves with thymus atrophy was significantly lower than that of control calves. TBX21 mRNA expression in peripheral CD4+ cells of thymus atrophy calves was tended to be higher than that of the control group. In addition, FOXP3 mRNA expression in peripheral CD4+ cells of the thymus atrophy calves was tended to be lower than that of the control calves. Thymus atrophy calves exhibited chronic inflammatory disease leading, in severe situations, to conditions such as pneumonia with caseous necrosis. These severe inflammatory responses likely are due to decreases in IL10 mRNA expression, impairing control of macrophages, one of the main cell fractions of natural immunity.  相似文献   

6.
This study was carried out to develop and characterize mouse monoclonal antibodies (mAbs) against chicken CD80 (chCD80). A recombinant plasmid containing a chCD80/horse IgG4 fusion gene was constructed and expressed in CHO cells to produce recombinant chCD80/IgG4 protein. Chicken CD80 was purified from the chCD80/IgG4 fusion protein following enterokinase digestion, and used to immunize BALB/c mice, resulting in 158 hybridomas that produced mAbs against chCD80. Three mAbs with high binding specificity for recombinant chCD80/IgG4-transfected CHO cells were identified by flow cytometry, and one of these (#112) was selected for further characterization. Immunoprecipitation of CD80/IgG4-CHO cell extract, or lipopolysaccharide (LPS)-treated monocytes identified 35.0 kDa proteins. Immunohistochemical analysis revealed chCD80-expressing cells exclusively in the bursal follicles at the outer portion of the cortex, and throughout the red pulp and the outer boundary of the white pulp in the spleen. By immunofluorescence microscopy, chCD80 was observed on intestinal dendritic cells. LPS treatment of bursa or spleen monocytes for 24 or 48 h increased chCD80 expression. Finally, addition of chCD80 mAb to Con A-stimulated spleen cells inhibited the expression of major histocompatibility complex class II antigens and IL-2-driven proliferation of lymphoblast cells. In summary, these chCD80 mAbs will serve as valuable immunological reagents for basic and applied poultry immunology research.  相似文献   

7.
Background: Tumors have heterogeneous properties, which could be explained by the existence of hierarchically and biologically distinct tumor cells such as tumor‐initiating cells (TICs). This model is clinically important, as TICs are promising targets for cancer therapies. However, TICs in spontaneous B‐cell lymphoma have not been conclusively identified. Hypothesis/Objectives: Tumor cells with a progenitor phenotype exist in B‐cell lymphoma, reflecting a hierarchical organization. Animals: Twenty‐eight client‐owned dogs with previously untreated B‐cell lymphoma and 6 healthy dogs. Methods: This was a prospective study. Flow cytometry was used to identify lymphoid progenitor cells (LPCs) that coexpressed hematopoietic progenitor antigens CD34, CD117, and CD133, with lymphoid differentiation markers CD21 and/or CD22 in B‐cell lymphoma. The polymerase chain reaction for antigen receptor rearrangements was used to analyze clonality and relatedness of tumor populations. A xenograft model with NOD/SCID/IL‐2Rγ?/? mice was adapted to expand and serially transplant primary canine B‐cell lymphoma. Results: LPCs were expanded in lymph nodes from 28 dogs with B‐cell lymphoma compared with 6 healthy dogs (P= .0022). LPCs contained a clonal antigen receptor gene rearrangement identical to that of the bulk of tumor cells. Canine B‐cell lymphoma xenografts in recipient mice that maintained LPCs in the tumors were recurrently observed. Conclusions and Clinical Importance: These results suggest the presence of a hierarchy of tumor cells in B‐cell lymphoma as has been demonstrated in other cancers. These findings have the potential to impact not only the understanding of lymphoma pathogenesis but also the development of lymphoma therapies by providing novel targets for therapy.  相似文献   

8.
Although a Yersinia pseudotuberculosis (Yptb) lung infection model has been developed to study Y. pestis pathogenesis, it is still necessary to establish a new animal model to mimic the pathophysiological features induced by Y. pestis infection. Here, we provide a new lung infection model using the Yptb strain, IP2777, which displayed rapid spread of bacteria to the liver, spleen, and blood. In addition, we examined whether TLR4 is involved in Yptb-induced pathogenesis in the lung infection model of mice we generated. Following lung infection of WT and TLR4-deficient mice with the Yptb strain IP2777, the survival rate, bacterial colonization, histopathology, and level of cytokines and chemokines in the lung, spleen, liver, and blood were analyzed. TLR4-deficient mice had a lower survival rate than WT mice in response to Yptb lung infection. Although the bacterial colonization and pathology of the lung were comparable between WT and TLR4-deficient mice, those of the spleen and liver were more severe in TLR4-deficient mice. In addition, the levels of TNF-α and CXCL2 in the liver and IL-6 and CXCL2 in the blood were higher in TLR4-deficient mice than in WT mice. Our results demonstrate that TLR4 is necessary for optimal host protection against Yptb lung infection and TLR4-deficient mice may serve as a better genetic model of Yptb infection for mimicking Y. pestis infection.  相似文献   

9.
The immunotropic properties of calf thymus extract (TFX-Jelfa) is connected with the mimic action of the thymus to modulate the differentiation, maturation and function of prothymocytes and mature thymus dependent (T) cells. The studies were carried out on CFW male mice aged 3 months. The animals were infected per os with 200 larvae of Trichinella spiralis. TFX-Jelfa was administered i.p. at a dose of 10 mg/kg seven times at 24 hour intervals prior to infection. The percentage of CD4+ and CD8+ in suspension of splenocytes and mesenteric lymphonode cells by flow cytometry using monoclonal antibodies coupled with fluorescein isothiocyanate (FITC) or phycoerythrin (PE) were determined. At the same time, cryostat preparations, made from jejunum and muscle samples, were examined by the direct immunofluorescence method using FITC-labeled antibody to mouse CD4+ and CD8+. It has been found that infection with T. spiralis in mice decreases the percentage of CD8+ splenocytes, while the percentage of CD8+ mesenteric lymphonode cells does not change. However, in infected mice the percentage of CD4+ spleen cells and mesenteric lymphonode cells is increased. It has been also found that during the course of infection an increase in the number of CD8+ and CD4+ cells in the basal lamina propria of the intestines was observed. In infected mice, CD4+ lymphocytes were visible in the inflammatory infiltrates of the muscle tissue on the 14th day, whereas CD8+ lymphocytes were first observed a week later. Pretreatment with TFX does not change the inhibitory effect of infection on the percentage of CD8+ splenocytes, but potentiates the percentage of CD4+ spleen cells and mesenteric lymphonode cells increased by infection. Furthermore, administration of TFX prior to infection also potentiates the stimulatory effect of T. spiralis on the number of CD8+ and CD4+ in the basal lamina propria of the jejunum, and on the number of CD8+ cells in the inflammatory infiltrates of the muscle tissue.  相似文献   

10.
Here, we report a case of spontaneous granulocytic leukemia in a 51-week-old male NOD/Shi-scid IL-2Rγnull (NOG) mouse. The mouse showed progressive anemia and rough respiratory movement. Macroscopically, the spleen was discolored and enlarged. Histologically, the bone marrow of the sternum and femur was highly cellular and almost exclusively filled with neoplastic cells. The nuclei of neoplastic cells were large, oval to slightly irregular in shape, and a small number of cells had kidney- or ring-shaped nuclei. Neoplastic cells extensively infiltrated the organs, and the spleen and liver were prominently involved. Immunohistochemically, a large population of neoplastic cells in the red pulp of the spleen and sinusoid of the liver was positive for myeloperoxidase. Based on the histological features, this case was diagnosed with granulocytic leukemia. This novel information on spontaneous tumors may be helpful for the appropriate use of this mouse strain in further research.  相似文献   

11.
Cryopreserved equine ocular squamous cell carcinoma (SCC) was inoculated subcutaneously into 15 athymic nude and 15 SCID mice. Xenotransplantation resulted in tumor growth in two athymic nude mice and 1 SCID mouse. Histological appearance and immunohistochemical characterization using cytokeratin 5/6 markers and p53 markers of the tumor grown in mice was in full accord with the original equine tumors. No evidence of metastasis was noted in any mouse. This model may serve as a relevant in vivo model for studying the biology of equine ocular SCC and for the testing of new therapeutic modalities.  相似文献   

12.
The Iowa strain of Cryptosporidium parvum will not propagate in immunocompetent mice, but will successfully infect genetically immunocompromised nude or SCID mice as well as immunocompetent mice which have been immunosuppressed with glucocorticoids. Using dexamethasone-tetracycline is one published method for immunosuppressing mice for the production of C. parvum oocysts. However, dexamethasone-induced immunosuppression is variable, because it is dependent on the total daily water consumption of each individual mouse. The changes in circulating leukocytes and other immune system associated organs before, during and after dexamethasone suppression were analyzed for comparison with a new single injection methylprednisolone acetate (MPA) suppression model. The dexamethasone-induced immunocompromised state was associated with a greater than 90% sustained drop in circulating T-lymphocytes, a greater than 700% increase in circulating mature segmented neutrophils and a severe depletion of circulating monocytes. The thymus and spleen decreased in size by over 80%. Oocyst shedding in suppressed mice started within 4 days of oocyst inoculation and persisted for 6 days post-dexamethasone treatment. Seven days after dexamethasone withdrawal, circulating neutrophils still were 549% higher than controls. Circulating CD3 and CD4 lymphocytes remained depressed by 85-90% while on dexamethasone and for 7 days after discontinuing dexamethasone. CD8 lymphocyte numbers initially decreased by 90%, but rose even while on dexamethasone and even with severe thymic involution. At day 7 post-dexamethasone treatment, the spleen was 119 mm(3), approximating the same size as controls. Fourteen days post-dexamethasone treatment, which was 8 days after oocyst shedding had ceased, the CD8 counts per 5000 events were only 1.6% below controls, while the CD3 and CD4 counts were still depressed by 66%. The thymus now was about one quarter smaller than the controls. The rise in circulating CD8 lymphocytes, when oocyst production stopped, suggests that CD8 positive lymphocytes may play a significant role in vivo in clearing the parasite. The overall pattern of immunosuppression was nearly identical to that observed with the methylprednisolone acetate immunosuppression model.  相似文献   

13.

Background

This study was performed to characterize a gene-addition transgenic mouse containing a BAC (bacterial artificial chromosome) clone spanning the human CYP2C18&19 genes (tg-CYP2C18&19).

Methods

Hemizygous tg-CYP2C18&19, 11 week old mice were compared with wild-type littermates to obtain information regarding clinical status, clinical pathology and anatomical pathology. After one week of clinical observations, blood samples were collected, organs weighed, and tissues collected for histopathology.

Results

In males, the tissue weights were lower in tg-CYP2C18&19 than in wild-type mice for brain (p ≤ 0.05), adrenal glands (p ≤ 0.05) and brown fat deposits (p ≤ 0.001) while the heart weight was higher (p ≤ 0.001). In female tg-CYP2C18&19, the tissue weights were lower for brain (p ≤ 0.001) and spleen (p ≤ 0.001) compared to wild-type females. Male tg-CYP2C18&19 had increased blood glucose levels (p ≤ 0.01) while females had decreased blood triglyceride levels (p ≤ 0.01).

Conclusion

Despite the observed alterations, tg-CYP2C18&19 did not show any macroscopic or microscopic pathology at the examined age. Hence, these hemizygous transgenic mice were considered to be viable and healthy animals.  相似文献   

14.
The outer membrane proteins (OMPs) of Brucella (B.) abortus have been extensively studied, but their immunogenicity and protective ability against B. abortus infection are still unclear. In the present study, B. abortus Omp28, a group 3 antigen, was amplified by PCR and cloned into a maltose fusion protein expression system. Recombinant Omp28 (rOmp28) was expressed in Escherichia coli and was then purified. Immunogenicity of rOmp28 was confirmed by Western blot analysis with Brucella-positive mouse serum. Furthermore, humoral- or cell-mediated immune responses measured by the production of IgG1 or IgG2a in rOmp28-immunized mice and the ability of rOmp28 immunization to protect against B. abortus infection were evaluated in a mouse model. In the immunogenicity analysis, the mean titers of IgG1 and IgG2a produced by rOmp28-immunized mice were 20-fold higher than those of PBS-treated mice throughout the entire experimental period. Furthermore, spleen proliferation and bacterial burden in the spleen of rOmp28-immunized mice were approximately 1.5-fold lower than those of PBS-treated mice when challenged with virulent B. abortus. These findings suggest that rOmp28 from B. abortus is a good candidate for manufacturing an effective subunit vaccine against B. abortus infection in animals.  相似文献   

15.
The studies were carried out on Balb/c mice exposed to restraint stress twice for 12 h at 24 h intervals. Prior to stress exposure, the mice were treated with calf thymus extract (TFX - Jelfa) i.p. at a dose of 10 mg/kg, ten times at 24 h intervals. TFX was used per se or with zinc ions interaction, by adding zinc ions (as sulfate salt) to drinking water at a dose of 72 microg/mouse per day. The results obtained show that restraint stress dramatically decreased the total number of thymocytes and splenocytes which is also accompanied by decreasing weight ratio of the thymus and spleen. The decreasing number of thymic and spleen cells corresponded to a diminishing percentage of immature, double-positive CD4+CD8+ thymocytes, mature single-positive CD4+ thymic cells and CD4+, CD8+ and CD19+ splenocytes. Changes in the number of thymic cells affect their activity, which is expressed as a decreased proliferative response of thymocytes stimulated in vitro with concanavalin A (Con A) and phytohaemagglutinin (PHA). Besides, exposure to the restraint stress decreased interleukin-1 (IL-1) production by murine intraperitoneal macrophages stimulated in vitro with lipopolisacharide (LPS) from E. coli. Previous treatment with TFX counteracted restraint stress-induced immunosuppression, which is expressed as partial normalisation of the total number of thymic and spleen cells, accelerated regeneration of these two lymphatic organs, shortned suppressive action of restraint stress on the percentage of immature CD4+CD8+ thymocytes and CD4+ splenocytes and in total normalisation of the CD4+ thymocytes and CD8+ splenocytes. TFX administered prior to restraint stress not only counteracted the suppresive effects of stress on the proliferative activity of thymic cells stimulated in vitro with Con A and PHA, but also augmented the proliferative response of these cells to two mitogens. The immunorestorative effect of TFX was augmented by zinc supplementation.  相似文献   

16.
In this study, we investigated the effects of oral ingestion of Lactobacillus crispatus KT‐11 strain (KT‐11) on the immune response in an allergic rhinitis mouse model, ovalbumin (OVA)‐sensitized BALB/c mice. Sneezing activity in mice that were administered a KT‐11‐supplemented diet was significantly lower than that in mice administered a KT‐11‐free diet (control diet) at age 11 weeks. We found that serum OVA‐specific immunoglobulin E (IgE) levels and total number of interleukin (IL)‐4+CD4+ spleen cells in mice that were administered a KT‐11‐supplemented diet were significantly lower than in mice administered a control diet. The ratio of spleen interferon‐γ+CD4+/IL‐4+CD4+ cells was higher in the mice administered the KT‐11‐supplemented diet compared to that in mice administered the control or L. rhamnosus GG‐supplemented diet. In contrast, the number of CD11b+CD80+ and FcεRIα+CD117+ cells was significantly lower in mice administered the KT‐11‐supplemented diet. These results suggested that KT‐11 reduced OVA‐induced allergic symptoms in BALB/c mice via the adjustment of the T helper type 1/T helper type 2 balance, and a decrease in the number of antigen‐presenting cells and high affinity IgE receptor‐positive mast cells.  相似文献   

17.
鼠源重组UBC13蛋白对脂多糖诱导的小鼠急性炎症的影响   总被引:1,自引:1,他引:0  
试验旨在探讨鼠源重组UBC13蛋白对脂多糖(lipopolysaccharide,LPS)诱导的小鼠急性炎症的影响。将24只SPF雌性小鼠随机分成4组:PBS组,LPS模型组,重组UBC13蛋白高、低剂量组(分别为100和25μg/只),每组6只。LPS模型组与各蛋白剂量组腹腔注射20 mg/kg LPS,PBS组腹腔注射等体积PBS;注射结束1 h后,各蛋白组按相应剂量背部皮下多点注射重组UBC13蛋白,PBS组与LPS模型组注射等体积PBS。给予蛋白24 h后处死小鼠。收集小鼠肺脏、脾脏、胸腺及肝脏组织,计算脏器指数,HE染色观察组织病理学变化,实时荧光定量PCR检测肺脏、脾脏、胸腺和肝脏中IL-1β、TNF-α、IL-6 mRNA的相对表达量,以及肺脏中iNOS mRNA的相对表达量,综合评价鼠源重组UBC13蛋白对LPS诱导小鼠急性炎症的影响。结果显示,与PBS组相比,LPS模型组小鼠肺脏、脾脏及肝脏指数均显著或极显著升高(P<0.05;P<0.01),且肺脏、脾脏和肝脏组织均出现病理变化。实时荧光定量PCR结果显示,与PBS组相比,LPS模型组肺脏、脾脏、胸腺和肝脏中IL-1β、TNF-α、IL-6 mRNA相对表达量均极显著升高(P<0.01),肺脏中iNOS mRNA相对表达量也极显著升高(P<0.01);与LPS模型组相比,UBC13蛋白高剂量组肺脏、脾脏和肝脏中病理变化明显改善,肺脏、肝脏、脾脏中IL-1β、TNF-α、IL-6及肺脏中iNOS mRNA表达量均极显著降低(P<0.01);胸腺中TNF-αmRNA表达量显著降低(P<0.05),IL-6 mRNA和IL-1β表达量极显著降低(P<0.01)。表明鼠源重组UBC13蛋白可下调炎性因子的表达,从而改善LPS诱导的小鼠急性炎症反应。  相似文献   

18.
Haemorrhagic septicaemia (HS) is an endemic disease of bovines, occuring in most tropical regions of Asia and Africa. In the present study, the suitability of using mice to study pathogenesis of HS was assessed using mortality, mean death time and bacterial multiplication in vital organs after infection with live P multocida. Mice were infected with 105, 103 and 101?cfu of P. multocida B:2 via intranasal and subcutaneous routes along with control groups. Bacterial multiplication in lung, liver and spleen of mice were determined at 24 h interval after intranasal and subcutaneous challenge. More than 80 % of challenged mice died within 48 h of inoculation, irrespective of the dose and route of inoculation. A heavy bacterial load (up to 108?cfu) was observed in lung, liver and spleen of mice titrated at 24 h and following death of mice. Results of the present study indicate that even ten bacteria are enough to cause mortality in mice and the organism multiplies rapidly in respiratory epithelium and disseminated to other vital organs viz liver and spleen suggesting the important role of mouse model in investigating the pathogenesis and challenge studies during vaccine development.  相似文献   

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研究茶树油提取物粉对小鼠免疫功能的影响,为茶树油产品的研发及作为畜禽养殖添加剂的应用积累资料。茶树油提取物粉对小鼠的经口急性毒性LD_(50)﹥5000 mg/kg b.w.。茶树油提取物粉分别以1250、250、50 mg/kg b.w.灌胃给予连续10 d,分别进行腹腔巨噬细胞吞噬活性和脾淋巴细胞增殖试验,测定免疫器官指数、血清溶血素水平及外周血CD3~+、CD4~+、CD8~+T淋巴细胞亚群水平。结果表明茶树油提取物粉高、中、低剂量组能极显著增加小鼠脾脏指数(P0.01),中剂量能显著增加肝脏指数和胸腺指数(P0.05)。中、高剂量能显著提高腹腔巨噬细胞吞噬活性和血清溶血素水平(P0.05)。三个剂量组均能显著提高小鼠脾淋巴细胞增殖的刺激指数(P0.05)。中剂量组能显著增加小鼠CD4+/CD8+比值(P0.05)。因此,茶树油提取物粉在一定程度上能够提高小鼠细胞免疫和体液免疫,具有增强小鼠免疫功能的作用。  相似文献   

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