Gamma-glutamyl transferase 5 overexpression in cerebrovascular endothelial cells improves brain pathology,cognition,and behavior in APP/PS1 mice

In patients with Alzheimer’s disease,gamma-glutamyl transferase 5(GGT5)expression has been observed to be downregulated in cerebrovascular endothelial cells.However,the functional role of GGT5 in the development of Alzheimer’s disease remains unclear.This study aimed to explore the effect of GGT5 on cognitive function and brain pathology in an APP/PS1 mouse model of Alzheimer’s disease,as well as the underlying mechanism.We observed a significant reduction in GGT5 expression in two in vitro models of Alzheimer’s disease(Aβ_(1-42)-treated hCMEC/D3 and bEnd.3 cells),as well as in the APP/PS1 mouse model.Additionally,injection of APP/PS1 mice with an adeno-associated virus encoding GGT5 enhanced hippocampal synaptic plasticity and mitigated cognitive deficits.Interestingly,increasing GGT5 expression in cerebrovascular endothelial cells reduced levels of both soluble and insoluble amyloid-βin the brains of APP/PS1 mice.This effect may be attributable to inhibition of the expression ofβ-site APP cleaving enzyme 1,which is mediated by nuclear factor-kappa B.Our findings demonstrate that GGT5 expression in cerebrovascular endothelial cells is inversely associated with Alzheimer’s disease pathogenesis,and that GGT5 upregulation mitigates cognitive deficits in APP/PS1 mice.These findings suggest that GGT5 expression in cerebrovascular endothelial cells is a potential therapeutic target and biomarker for Alzheimer’s disease.

Ketogenic diet alleviates cognitive dysfunction and neuroinflammation in APP/PS1 mice via the Nrf2/HO-1 and NF-κB signaling pathways

Alzheimer's disease is a progressive neurological disorder characterized by cognitive decline and chronic inflammation within the brain.The ketogenic diet,a widely recognized therapeutic intervention for refractory epilepsy,has recently been proposed as a potential treatment for a variety of neurological diseases,including Alzheimer's disease.However,the efficacy of ketogenic diet in treating Alzheimer's disease and the underlying mechanism remains unclear.The current investigation aimed to explore the effect of ketogenic diet on cognitive function and the underlying biological mechanisms in a mouse model of Alzheimer's disease.Male amyloid precursor protein/presenilin 1(APP/PS1)mice were randomly assigned to either a ketogenic diet or control diet group,and received their respective diets for a duration of 3 months.The findings show that ketogenic diet administration enhanced cognitive function,attenuated amyloid plaque formation and proinflammatory cytokine levels in APP/PS1 mice,and augmented the nuclear factor-erythroid 2-p45 derived factor 2/heme oxygenase-1 signaling pathway while suppressing the nuclear factor-kappa B pathway.Collectively,these data suggest that ketogenic diet may have a therapeutic potential in treating Alzheimer's disease by ameliorating the neurotoxicity associated with Aβ-induced inflammation.This study highlights the urgent need for further research into the use of ketogenic diet as a potential therapy for Alzheimer's disease.

Carboxymethytl Pachymaram Up-Regulates Dendritic Cell's Function in Hepatitis B Virus Transgenic Mice in vitro

The effect ofcarboxymethytl pachymaram （ CMP ） on the function of dendritic cells（DCs） derived from spleens of hepatitis B virus transgenic mice are studied in vitro. The phenotypes of DCs are tested by flow cytometry （FCM）, cytokines measured by ELISA. The expression of DCs＇ phenotypes in IdBV transgenic mice are low （CD80^＋CD11c^＋：59.12±11.53 vs 9,60±4.53, p〈0.01; CD80^＋ MHC-Ⅱ^＋： 44.86±12.31 vs 9.80±5,72, p〈0.01, normal mice vs HBV transgenic mice）, the ability of DCs stimulating T lymphocytes proliferation decreases （0.37±0.11 vs 0.20±0,11, p〈0.05, normal mice vs HBV transgenic mice）, levels of IL-12 and IFN-y decrease whereas the level of IL-10 increases; CMP can enhance DCs＇ ability of stimulating T lymphocytes proliferation, facilitate the secretion of IL-12 and IFNp, inhibit the secretion of IL-10, thus up regulates DCs function. The results show a good prospective use of CMP on the treatment of chronic hepatitis B.

Shuanghuanglian injection downregulates nuclear factor-kappa B expression in mice with viral encephalitis

A mouse model of viral encephalitis was induced by intracranial injection of a Coxsackie virus B3 suspension. Quantitative real-time reverse transcription-PCR and western blot assay were applied to detect mRNA and protein expression of intelectin-2 and nuclear factor-kappa B in the viral encephalitis and control groups. Nuclear factor-kappa B and intelectin-2 mRNA and protein expression were significantly increased in mice with viral encephalitis. After intraperitoneal injection of Shuanghuanglian at a dose of 1.5 mg/kg for 5 successive days, intelectin-2 and nuclear factor-kappa B protein and mRNA expression were significantly decreased. To elucidate the relationship between intelectin-2 and nuclear factor-kappa B, mice with viral encephalitis were administered an intracerebral injection of 107 pfu recombinant lentivirus expressing intelectin shRNA. Both protein and mRNA levels of intelectin and nuclear factor-kappa B in brain tissue of mice were significantly decreased. Experimental findings suggest that Shuanghuanglian injection may downregulate nuclear factor-kappa B production via suppression of intelectin production, thus inhibiting inflammation associated with viral encephalitis.

Antiviral Effects of Stichopus japonicus Acid Mucopolysaccharide on Hepatitis B Virus Transgenic Mice

Hepatitis B virus(HBV) is a significant global pathogen and efficient cure for HBV patients is still a challenging goal. We previously reported that acidic mucopolysaccharide from stichopus japonicus selenka(SJAMP) could inhibit HBs Ag and HBe Ag expression in vitro. However, the potential anti-HBV effects of SJAMP in vivo have not yet been explored. In this study, we show that SJAMP exhibits potent anti-HBV activity in HBV transgenic mice in a dose-dependent manner. Specifically, sixty HBV transgenic male BALB/c mice were randomly selected to receive the treatment of PBS, low dose SJAMP(30 mg kg^(-1)), middle dose SJAMP(40 mg kg^(-1)), high dose SJAMP(50 mg kg^(-1)) and IFN(45 IU kg^(-1)) for 30 d. SJAMP treatment suppressed serum HBV-DNA, and liver HBs Ag and HBc Ag levels in HBV-transgenic mice. The present study highlights the potential application of SJAMP in HBV therapy.

Global view of transcriptome in the brains of aged NR2B transgenic mice

NR2B subunits are involved in regulating aging, in particular, age-related learning and memory deficits. We examined 19-month-old NR2B transgenic mice and their littermate controls. First, we detected expression of the NR2B subunit gene, Grin2b, in the neocortex of transgenic mice using real-time PCR. Next, we used microarrays to examine differences in neocortical gene expression. Pathway and signal-net analyses identified multiple pathways altered in the transgenic mice, in-cluding the P53, Jak-STAT, Wnt, and Notch pathways, as well as regulation of the actin cytoskeleton and neuroactive ligand-receptor interactions. Further signal-net analysis highlighted the P53 and insulin-like growth factor pathways as key regulatory pathways. Our results provide new insight into understanding the molecular mechanisms of NR2B regulated age-related memory storage, normal organismal aging and age-related disease.

Effects of Langchuangjing Granule (狼疮净冲剂) on Apoptotic of CD_4^+T and CD_(19)^+B in Spleen of BXSB Mice with Systemic Lupus Erythematosus

Objective: To study the effect on apoptosis of CD 4 +T、CD 19 +B in spleen of BXSB mice with systemic lupus erythematosus treated with Langchuangjing Granule (LCJG, and to probe into the mechanism of the tr eatment. Methods: The apoptosis was examined by the flow cytometric analysis and immunofluorescence double-staining method. Results: Apoptosis of male BXSB mice speeds up. LCJG can restrain the excessive apoptosis of CD 4 +T and CD 19 +B cells in spleen. Conclusion: LCJG treated systemic lupus erythematosus by restraining the excessive apoptosis of T, B lymphocytes, probably restraining the release of excessive amount of apoptotic DNA fragments, so decreasing abnormal proliferation of B cells and the produce of autoantibodies.

Detraining after tumor-bearing accelerates tumor growth while continuous training decreases tumor growth in mice

Objective:Previous studies have shown that exercise suppresses tumor growth.However,the effects of exercise with different intensities and exercise detraining after tumor-bearing on tumor progression remain unclear.The purpose of this study was to investigate the effects of continuous and disrupted free and exhausted swimming training after tumor-bearing on tumor progression in melanoma B16-F10-bearing C57BL/6 mice.Methods:C57BL/6 mice were subjected to free or exhausted swimming exercise training for 4 weeks prior to the injection of melanoma B16-F10 cells.Subsequently,the B16-F10-bearing mice were maintained with training consisting of free or exhausted swimming or without exercise for 2 weeks during the tumor challenge.Results:The tumor weight was increased by 42%and 109%in mice with 4-week exhausted swimming prior to B16-F10 tumor cells inoculation followed by 2-week training cessation compared with the tumor-bearing control(P<.05)and continuous training groups(P<.01).Tumor weights in groups with exercise detraining after tumor cell inoculation tended to be increased,while the proliferation of splenic T lymphocytes tended to be decreased compared with the group that maintained exercise intensity.After 6-weeks continuous free or exhausted swimming training,the tumor weight of mice was decreased and the proliferation of splenic T lymphocytes was increased compared with the tumor-bearing control group.The frequency of natural killer cells in tumors was increased in all exercise training groups of mice.Conclusions:These results suggest that maintaining exercise intensity after tumor-bearing slows tumor growth in mice,possibly because of the enhanced proliferative activity of splenic lymphocytes rather than natural killer cell infiltration.However,detraining after tumor-bearing might accelerate tumor progression because of the reduced proliferation of splenic T lymphocytes.

The involvement of IL-23/Th17 pathway in mice virus myocarditis inducedby cosakievirus B3

Background The IL-23/ Th17 pathway plays an important role in the development of chronic in? ammatory diseases and autoimmune diseases.However,the role of the IL-23/Th17 axis in the regulation of virus myocarditis is still largely unknown.We aim to determine the role of IL-23/ Th17 axis in virus myocartidis.Methods and Results Balb/c male mice were peritoneally injected with 100TICD50 Coxsackie virus B3 to establish virus myocarditis(VMC), mice injected with PBS peritoneally were taken as the controls. 0,1,2,3,4 and 6 weeks After injection,IL-23,L-17 and RORγt mRNA in the myocardium of VMC were assessed by Semi-quantitative RT-PCR,and IL-23 protein from plasm was evaluated by ELISA.Flow cytometric analysis was used to evaluate the frequencies of Th17 subsets in CD4,To investigated whether the IL-23 is important during IL-23/Th-17 pathway challenge,we isolated CD4 + T cells and cultured with rIL-23 in vitro,and examined the Th17 cells.Results show that,comparing with the controls,IL-23,IL-17 and RORγt mRNA was steadly expressed in the myocardiums of infected mice from 1 week after virus injection.Conclusions IL-23/ Th-17 pathway may therefore play an essential role in VMC. Comparative studies are required to reveal further the roles of these cytokines in the pathogenesis of? these immune-related diseases.

The cloning of 3'-truncated preS/S gene from HBV genomic DNA and its expression in transgenic mice

INTRODUCTIONHepatitis B virus (HBV) is regarded as one of themain etiologic factors involved in the developmentof human hepatocellular carcinoma (HCC).The open reading frame (orf)of X gene of HBVencoded a transactivating factor is the evidence thatstrongly supported the notion that the X gene ofHBV DNA integrated in HCC genomic DNA couldcontribute to the carcinogenesis of liver cells byactivation of some related cellular genes

Sex disparity in viral load, inflammation and liver damage in transgenic mice carrying full hepatitis B virus genome with the W4P mutation in the preS1 region

AIM To study sex disparity in susceptibility to hepatocellular carcinoma(HCC), we created a transgenic mouse model that expressed the full hepatitis B virus(HBV) genome with the W4P mutation.METHODS Transgenic mice were generated by transferring the p HY92-1.1 x-HBV-full genome plasmid(genotype A2) into C57 Bl/6 N mice. We compared serum levels of hepatitis B surface antigen(HBs Ag), interleukin(IL)-6, and the liver enzymes alanine aminotransferase(ALT) and aspartate transaminase(AST), as well as liver histopathological features in male and female transgenic(W4PTG) mice and in nontransgenic littermates of 10 mo of age. RESULTS W4PTG males exhibited more pronounced hepatomegaly, significantly increased granule generation in liver tissue, elevated HBs Ag expression in the liver and serum, and higher serum ALT and IL-6 levels compared to W4PTG females or littermate control groups. CONCLUSION Together, our data indicate that the W4 P mutation in pre S1 may contribute to sex disparity in susceptibility to HCC by causing increased HBV virion replication and enhanced IL-6-mediated inflammation in male individuals. Additionally, our transgenic mouse model that expresses full HBV genome with the W4 P mutation in pre S1 could be effectively used for the studies of the progression of liver diseases, including HCC.

Evaluating the immune responses of mice to subcutaneous immunization with Helicobacter pylori urease B subunit

Background： Helicobacter pylori, a gram-negative bacterial pathogen that expresses a strong urease activity, is associated with the development of gastroduodenal disease. Urease B subunit, one of the two structural subunits of urease, was expressed in E. coil BL21 （DE3） strain. The objective of this study was to evaluate the effects of He/icobacter py/ori urease B subunit on the immune responses in mice by subcutaneous immunization. Methods： The mice were immunized and boosted with Helicobacter pylori urease B subunit antigen subcutaneously three times with 2-wk intervals between the immunizations and boosters. The mice in the control group were immunized with PBS. The adjuvant group received PBS containing complete/incomplete freund＇s adjuvant identical to antigen group without Helicobocter pylori urease B subunit antigen. Four weeks after the final booster, all the mice were sacrificed. Blood was collected on d 0, 14, 28 and 56 before immunization, booster and sacrifice, respectively. Immediately after sacrifice, gastric liquid and spleen were collected for antibody and cytokine analyses. Results： Urease B subunit increased the concentrations of serum and gastric anti-urease B antigen specific IgG, and the levels of interteukin-4 and interferon-y in splenocytes of the mice （P 〈 0.05）. Conclusions： This study demonstrated that recombinant responses in mice by subcutaneous immunization, which against Helicobocter pylori. urease B subunit can induce systemic and local immune might be used as the effective component of vaccine

Antiviral effects of hepatitis B virus S gene-specific anti-gene locked nucleic acid in transgenic mice

AIM To assess the antiviral effects of hepatitis B virus(HBV) S gene-specific anti-gene locked nucleic acid(LNA) in transgenic mice.METHODS Thirty HBV transgenic mice were acclimatized to laboratory conditions and positive for serum HBV surface antigen(HBs Ag) and HBV DNA, were randomly divided into 5 groups(n = 7), including negative control(blank control, unrelated sequence control), positive control(lamivudine, anti-sense-LNA), and anti-gene-LNA experimental group. LNA was injected into transgenic mice by tail vein while lamivudine was administeredby gavage. Serum HBV DNA and HBs Ag levels were determined by fluorescence-based PCR and enzymelinked immune sorbent assay, respectively. HBV S gene expression amounts were assessed by reverse transcription polymerase chain reaction. Positive rates of HBsA g in liver cells were evaluated immunohistochemistry.RESULTS Average rate reductions of HBs Ag after treatment on the 3 rd, 5 th, and 7 th days were 32.34%, 45.96%, and 59.15%, respectively. The inhibitory effect of antigene-LNA on serum HBs Ag peaked on day 7, with statistically significant differences compared with pretreatment(0.96 ± 0.18 vs 2.35 ± 0.33, P < 0.05) and control values(P < 0.05 for all). Average reduction rates of HBV DNA on the 3 rd, 5 th, and 7 th days were 38.55%, 50.95%, and 62.26%, respectively. This inhibitory effect peaked on the 7 th day after treatment with anti-gene-LNA, with statistically significant differences compared with pre-treatment(4.17 ± 1.29 vs 11.05 ± 1.25, P < 0.05) and control values(P < 0.05 for all). The mR NA levels of the HBV S gene(P < 0.05 for all) and rates of HBsA g positive liver cells(P < 0.05 for all) were significantly reduced compared with the control groups. Liver and kidney function, and histology showed no abnormalities. CONCLUSION Anti-gene-LNA targeting the S gene of HBV displays strong inhibitory effects on HBV in transgenic mice, providing theoretical and experimental bases for gene therapy in HBV.

Aralia elata inhibits neurodegeneration by downregulating O-GlcNAcylation of NF-κB in diabetic mice

AIM： To investigate the role of O-GIcNAcylation of nuclear factor-kappa B （NF-KB） in retinal ganglion cell （RGC） death and analysedthe effect of Aralia elata （AE） on neurodegen- eration in diabetic mice. METHODS： C57BL/6mice with streptozotocin-induced diabetes were fed daily with AE extract or control （CTL） diet at the onset of diabetes mellitus （DM）. Two months af- tar injection of streptozotocin or saline, the degree of cell death and the expression of O-GIcNAc transferase （OGT）, N-acetyl-b-D-glucosaminidase （OGA）, O-GIcNAcylated pro- teins, and O-GIcNAcylation of NF-KB were examined. RESULTS： AE did not affect the metabolic status of diabetic mice. The decrease in the inner retinal thickness （P〈0.001 vs CTL, P〈0.01 vs DM） and increases in RGCs with terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling （P〈0.001 vs CTL, P〈0.0001 vs DM）, glial activation, and active caspase-3 （P〈0.0001 vs CTL, P〈0.0001 vs DM） were blocked in diabetic retinas of AE extract-fed mice. Expression levels of protein O-GIcNAcylation and OGT were increased in diabetic retinas （P〈0.0001 vs CTL）, and the level of O-GIcNAcylation of the NF-KB p65 subunit was higher in diabetic retinas than in controls （P〈0.0001 vs CTL）. AE extract downregulated O-GIcNAcylation of NF-KB and prevented neurodegeneration induced by hyperglycemia （P〈0.0001 vs DM）. CONCLUSION： O-GIcNAcylation of NF-KB is concerned in neuronal degeneration and that AE prevents diabetes-in- duced RGC apoptosis via downregulation of NF-KB O-GI- cNAcylation. Hence, O-GIcNAcylation may be a new object for the treatment of DR, and AE may have therapeutic pos- sibility to prevent diabetes-induced neurodegeneration.

Metabolomic Study on Vitamins B_1, B_2, and PP Supplementation to Improve Serum Metabolic Profiles in Mice under Acute Hypoxia Based on ~1H NMR Analysis

Objective To explore metabolic changes after acute hypoxia and modulating effect of vitamins B1, B2, and PP supplementation in mice exposed to acute hypoxia. Methods Fifty male Kunming mice were randomly divided into 5 groups： normal, acute hypoxia, acute hypoxia with 2, 4 and 8 time- vitamins B1, B2, and PP supplementation . All mice were fed with corresponding diets for two weeks and then were exposed to a simulated altitude of 6 000 meters for 8 h, except for the normal group. Nuclear magnetic resonance analysis was used to identify the changes of serum metabolic profiles. Results There were significant changes in some serum metabolites under induced acute hypoxia, essentially relative increase in the concentrations of lactate, sugar and lipids and decrease in ethanol. The serum levels of choline, succinate, taurine, alanine, and glutamine also increased and phosphocholine decreased in the acute hypoxia group. After vitamins B1, B2, and PP supplementation, all these metabolic changes gradually recovered. Conclusion Significant changes in serum metabolic profile were observed by metabolomics in mice exposed to acute hypoxia, and vitamins B1, B2, and PP supplementation proved to be beneficial to improving some metabolic pathways. It is suggested that the dietary intakes of vitamins B1, B2, and PP should be increased under hypoxia condition.

Inhibitory effects of Shuanghuanglian injection on nuclear factor-kappa B expression in mice with viral encephalitis in a time-and dose-dependent manner

Previous studies have confirmed that the anti-virus effects of Shuanghuanglian injection may be associated with nuclear factor-kappa B activity. This study observed nuclear factor-kappa B expression in mice with viral encephalitis, and showed significant decreases in nuclear factor-kappa B protein and mRNA levels following Shuanghuanglian injection. The inhibitory effect was more significant with prolonged intervention duration and increased treatment dose. These findings verify that Shuanghuanglian injection plays a therapeutic role in viral encephalitis by reducing expression of nuclear factor-kappa B in a time- and dose-dependent manner.

Study on the production of IL-12, IL-10 in early stage after infection with Babesia microti and Babesia rodhaini in mice

The serum levels of IL-12 and IL-10 in mice after infected with Babesia microti （B. microti） and Babesia rodhaini （B. rodhaini） were examined. Collected the mice serum and examined the concentration of IL-12 and IL-10 by using ELISA after infection with B. microti and B. rodhaini at 0, 3, 6, 9, 12, 18, 24, 36, 72, 96 h in mice. The results showed that B. microti infection resulted in IL-12 increasing, which peaked at 3 and 24 h after the infection, while same infection did not induce a significant change in IL-10 compared to uninfected mice. When mice were infected with B. rodhaini, any significant changes were not decteted both in IL-12 and IL-10 in comparison with uninfected animals during the period of 3-72 h after infection. Instead, a significant decline in IL-12 and IL-10 was found compared to uninfected mice 96 h after infection with B. rodhaini. It indicates that the mutagenetic cytokine is IL-12 in the serum of mice after infection with B. microti, and no any significant changes were detected in both IL-12 and IL-10 from 0 to 72 h after infected with B. rodhaini.

Antiviral treatment of hepatitis B virus-transgenic mice by a marine organism, Styela plicata

瞄准：在肝炎 B 搬运人的一个鼠科的模型评估 Styela plicata 的有效成分的抗病毒的效果。方法：HBV 转基因的老鼠被划分成 3 个组(控制组， lamivudine 处理组和 Styela plicata 处理组的有效成分) 并且分配了收到正常饮食， lamivudine 或 Styela plicata 的有效成分连续星期。浆液肝炎 B 表面抗原被连接酶的免疫检测吸着剂试金(ELISA ) 方法。浆液 HBV DNA 被即时聚合酶链反应(RT-PCR ) 检测。浆液 T 助手(h) 1 cytokine interleukin (IL )-2 和 Th2 cytokine IL-6 被量的三明治酶免疫分析技术检测。另一组 HBV 转基因的老鼠被分配收到 Styela plicata 的有效成分连续星期。肝组织的组织学在治疗前后被评估。结果：12 个星期在开始治疗以后，浆液肝炎 B 表面抗原显著地在 Styela plicata 被降低与收到正常饮食的老鼠相比对待老鼠和对待 lamivudine 的老鼠(F (12wk )= 88.81， P (12wk )= 0.000 【0.01 ) 。浆液 HBV DNA 显著地在 Styela plicata 被降低与收到正常饮食的老鼠相比对待老鼠和对待 lamivudine 的老鼠(F (12wk )= 20.71， P (12wk )= 0.000 【0.01 ) 。然而， Styela plicata 的有效成分象 lamivudine 一样，不能完全禁止 HBV 的复制。在重量的单位的肝炎 B 表面抗原和 HBV DNA 的回缩现象一 could 在药的退却以后被发现 4 wk。八个星期在开始治疗以后，在 IL-2 的 Styela plicata 处理前后的浆液层次是 2.41 +/- 0.38 和 10.56 +/- 0.78 ng/L，分别地(t (8wk )=-16.51, P (8wk )= 0.000 【0.01 ) 。与在正常对待食谱的老鼠的 IL-2 的浆液层次相比(2.48+/-0.17 ng/L；t (8wk )= 13.23， P (8wk )= 0.000 【0.01 ) 。在 IL-6 的 Styela plicata 处理前后的浆液层次是 63.62 +/- 分别地， 6.22 ng/L 在正常对待食谱的老鼠 IL-6 与浆液相比铺平的 6.31 和 54.52 +/-(60.84 +/- 4.21 ng/L ) 。从 Styela 对待 plicata 的 HBV 转基因的老鼠的肝的组织学的分析也在发炎和肝炎 B 表面抗原显示出下降地位。结论：Styela plicata 可以是在对待长期的肝炎 B 的有效的抗病毒的药。

Hepatitis B virus x gene and cyanobacterial toxins promote aflatoxin B_1-induced hepatotumorigenesis in mice

瞄准：估计黄麴毒素的结合的角色 B1 (AFB1 ) ， cyanobacterial 毒素(cyanotoxins ) ，和肝炎 B (HBV ) 在 hepatotumorigenicity 的 x 基因。方法：( ip )带 HBV x 基因和他们的野类型的同窝出生的人的 One-week-old 动物 intraperitoneally 与也被注射单个剂量的 AFB1 [ 6 mg/kg 体重( bw )]，重复剂量 cyanotoxins ( microcystin-LR 或在榴状， 10 microg/kg bw 每周一次为 15 wk )， DMSO (车辆控制)独自一个，或 AFB1 一个星期以后由 cyanotoxins 列在后面，并且在 24 和 52 wk 被牺牲处理以后。结果：AFB1 在 29 中的 13 个导致了肝肿瘤(44.8%) 在 52 wk 的转基因的老鼠处理以后，显著地比在野类型的老鼠(13.3%) 更经常。这有效差量没在 24-wk 学习被显示出。与 AFB1 暴露相比独自一个， MC-LR 并且在榴状在导致的 AFB (1 ) 的发生产出近似 3 褶层和 6 褶层增加在在 24 wk 的野类型的动物的肝肿瘤分别地。HBV x 基因进一步没提高与合作暴露联系到 AFB1 和 cyanotoxins 的风险。与一个 MC-LR-dosed 野类型的鼠标的异常，没有肝肿瘤在在 24 wk 独自与 cyanotoxins 对待的鼠标被观察。既不对待 DMSO 的转基因的老鼠也不他们的野类型的同窝出生的人在甚至多达 52 wk 有与 hepatotumorigenesis 相关的病理学的改变。结论：HBV x 基因并且在榴状支持导致的 AFB (1 ) 的发展肝肿瘤。到 AFB1 和 MC-LR 的合作暴露趋于相对暴露在 24 wk 提高肝肿瘤的风险到他们之一。hepatotumorigenesis 上的 AFB1， cyanotoxins 和 HBVx 的结合的效果在 24 wk 是弱的。