Activation of Akt signal pathway cascades in kidney tissue in murine chronic graft-versus-host disease lupus nephritis and its regulation by prednisone

AIM:To examine whether Akt signal pathway proteins, including Akt, NF-κB and IκBα, are activated in kidney tissue of murine chronic graft-versus-host disease (GvHD) lupus nephritis in vivo, and whether prednisone suppresses activation of them.METHODS:Akt activity and phosphorylated IκBα were detected by Western-blot. Activation of NF-κB was detected by electropheretic mobility shift assay (EMSA). RESULTS:Activity of Akt, NF-κB and phosphorylated IκBα were significantly increased in kidney tissue of murine chronic graft-versus-host disease (GvHD) in 8th week and 12th week after monocell injection, respectively. However, they were no significant elevation in 16th week, when compared with controls. Prednisone treatment significantly prevented the increase in serum anti-dsDNA antibody level, urinary protein excretion and glomerular cell proliferation in GvHD mice, indicating the beneficial effects of prednisone on this model. Prednisone also significantly suppressed the increase in the activities of glomerular Akt, NF-κB and phosphorylated IκBα. CONCLUSION:This study provides the first evidence of marked increase in glomerular Akt-NF-κB signal pathway activities in murine chronic graft-versus-host disease lupus nephritis. The beneficial effect of prednisone on this lupus nephritis model may be partially mediated by the suppression of abnormal Akt- NF-κB activation.     

Hexokinase 2 inhibits LPS-induced mitochondria-dependent apoptosis in human lung epithelial BEAS-2B cells

AIM: To explore the effect of hexokinase 2 (HK2) on lipopolysaccharide (LPS)-induced apoptosis of human lung epithelial BEAS-2B cells and the underlying mechanisms.METHODS: BEAS-2B cells were treated with LPS to induce cell injury, and the cell viability was examined by CCK-8 assay. Hoechst 33342 staining and Annexin V/PI double staining were used to analyze the apoptosis. The apoptotic pathway was identified by the specific inhibitor for caspase-8 or caspase-9. The releases of key mediators in mitochondrial apoptosis pathway were examined by Western blot. The effects of HK2 in these process were confirmed by HK2 over-expression followed by LPS treatment.RESULTS: CCK-8 assay showed that LPS treatment decreased the viability of BEAS-2B cells in a dose/time-dependent manner (P<0.01). The apoptosis of BEAS-2B cells was manifested by Hoechst 33342 and Annexin V/PI double staining. Pretreatment with z-LEHD-fmk, but not z-IETD-fmk, reversed the decreased cell viability under LPS stimulation. HK2 down-regulation was involved in LPS-induced apoptosis of the BEAS-2B cells. After HK2 over-expression, the cell viability was increased after LPS treatment. Releases of cytochrome C and apoptosis-inducing factor from mitochondrion to cytoplasm during apoptosis were also inhibited by HK2 over-expression.CONCLUSION: Hexokinase 2 inhibits LPS-induced mitochondria-dependent apoptosis in human lung epithelial cells.     

In vivo control and in vitro antifungal activity of rhamnolipid B,a glycolipid antibiotic,against Phytophthora capsici and Colletotrichum orbiculare

The glycolipid antibiotic rhamnolipid B isolated from Pseudomonas aeruginosa strain B5 was evaluated for in vitro antifungal activity and in vivo control against phytophthora blight and anthracnose under glasshouse conditions. Rhamnolipid B showed antifungal activity against Cercospora kikuchii, Cladosporium cucumerinum, Colletotrichum orbiculare, Cylindrocarpon destructans, Magnaporthe grisea and Phytophthora capsici. Microscopic observation revealed that the high level of antifungal activity (10 µg ml ⁻¹) against P capsici was mainly due to a lytic effect on zoospores. Zoospore lysis began in the presence of 10 µg ml ⁻¹ of rhamnolipid B and most of the zoospores were collapsed at 25 µg ml ⁻¹. Rhamnolipid B showed inhibitory activity against the germination of zoospores and hyphal growth of P capsici at concentrations of 50 µg ml ⁻¹. Spore germination of the anthracnose plant pathogen C orbiculare was also inhibited in the presence of 50 µg ml ⁻¹ of rhamnolipid B, although hyphal growth was not affected at this concentration. In the glasshouse, the efficacy of rhamnolipid B against phytophthora blight was similar to that of metalaxyl on pepper plants when treated just before inoculation with P capsici. Treatment with either at 500 µg ml ⁻¹ completely protected pepper plants from phytophthora blight. Rhamnolipid B also suppressed the development of C orbiculare infection on leaves of cucumber plants. © 2000 Society of Chemical Industry     

Chitosan Oligosaccharides Alleviate Colitis by Regulating Intestinal Microbiota and PPARγ/SIRT1-Mediated NF-κB Pathway

Chitosan oligosaccharides (COS) have been shown to have potential protective effects against colitis, but the mechanism underlying this effect has not been fully elucidated. In this study, COS were found to significantly attenuate dextran sodium sulfate-induced colitis in mice by decreasing disease activity index scores, downregulating pro-inflammatory cytokines, and upregulating Mucin-2 levels. COS also significantly inhibited the levels of nitric oxide (NO) and IL-6 in lipopolysaccharide-stimulated RAW 264.7 cells. Importantly, COS inhibited the activation of the NF-κB signaling pathway via activating PPARγ and SIRT1, thus reducing the production of NO and IL-6. The antagonist of PPARγ could abolish the anti-inflammatory effects of COS in LPS-treated cells. COS also activated SIRT1 to reduce the acetylation of p65 protein at lysine 310, which was reversed by silencing SIRT1 by siRNA. Moreover, COS treatment increased the diversity of intestinal microbiota and partly restored the Firmicutes/Bacteroidetes ratio. COS administration could optimize intestinal microbiota composition by increasing the abundance of norank_f_Muribaculaceae, Lactobacillus and Alistipes, while decreasing the abundance of Turicibacte. Furthermore, COS could also increase the levels of propionate and butyrate. Overall, COS can improve colitis by regulating intestinal microbiota and the PPARγ/SIRT1-mediated NF-κB pathway.     

Effect of KLF4 on viability,apoptosis of colorectal cancer cells and chemosensitivity to cisplatin

AIM:To investigate the effect of Krüppel-like factor 4 (KLF4) on the viability, apoptosis and cisplatin chemosensitivity of colorectal cancer cells. METHODS:KLF4 expression in colorectal cancer cell lines Caco2, SW480 and HCT116 was detected by Western blot. The SW480 cells were divided into pcDNA3.1 group (transfected with pcDNA3.1 empty plasmid), pcDNA3.1-KLF4 group (transfected with pcDNA3.1-KLF4 expression plasmid) and pcDNA3.1-KLF4+cisplatin group (treated with 1 mg/L cisplatin for 48 h after pcDNA3.1-KLF4 was transfected into SW480 cells). The protein levels of KLF4, p-IκBα, cyclin D1 and survivin were determined by Western blot. The cell viability was measured by CCK-8 assay. The apoptotic rate was analyzed by flow cytometry. The content of reactive oxygen species(ROS) was measured by DCFH-DA probe. RESULTS:The expression of KLF4 in the colorectal cancer cells were significantly lower than that in the human colon mucosal epithelial NCM460 cells (P<0.05). Compared with pcDNA3.1 group, the protein expression of KLF4 in pcDNA3.1-KLF4 group was significantly increased (P<0.05). Compared with pcDNA3.1 group, the cell viability and the protein expression of cyclin D1 and survivin were significantly decreased, and the apoptotic rate, the content of ROS and the protein level of p-IκBα were significantly increased in pcDNA3.1 group (P<0.05). Compared with pcDNA3.1-KLF4 group, the cell viability and the expression of cyclin D1 and survivin proteins were significantly decreased, and the apoptotic rate, the content of ROS and the protein level of p-IκBα were significantly increased in pcDNA3.1-KLF4+cisplatin group (P<0.05). CONCLUSION:Upregulation of KLF4 gene expression in colorectal cancer cells reduces the cell viability, induces apoptosis and increases the chemosensitivity of the cells to cisplatin. The mechanism may be related to the enhancement of intracellular ROS content and down-regulaton of the phosphorylation level of IκBα, the key molecule of NF-κB signaling pathway.     

甘蓝型油菜（B.napusL.）细胞核雄性不育系杂种“中杂3号”的选育和遗传分析

以70-506花叶自交系中的天然雄性不育株为母本,先后同齐菲和加油3号杂交以改良综合经济性状。再连续用兄妹交保持和选择,育成1081AB雄性核不育“两用系”。通过不同品种测交和选育,育成恢复系1081C。在组合鉴定试验中,F1杂种比CK(甘油5号)增产30％;在湖北省油菜区试中比“中油821”增产11.45％,含油量高3.92％,成熟早3.8天。并对1081AB不育系的遗传组成作了初步分析。     

春秋兼用蚕品种57A·57B、24·46的育成及其四元杂交种的选配

采用杂交育种方法,在春、夏、秋不同环境下定向培育,经严格的后代选择,育成二个中系限性品种57A、57B和二个日系品种24、46,然后运用杂种强势的原理,选配成四元杂交种57A·57B×24·46.经多次鉴定结果表明,该四元杂交种的产量、产值比对照种提高3—4%;且体质强健,对叶质要求不高,制种较容易,丝质优良,是春秋兼用的优良蚕品种.     

芸芥(Eruca sativa Mill.)对菌核病的抗性研究

 经对芸芥和甘蓝型油菜进行苗期菌丝块接种鉴定和花期带菌牙签鉴定表明,芸芥品种总体抗性水平高于甘蓝型油菜。所鉴定的32个芸芥品种中,抗病品种22个,占供试品种68.8％,感病品种10个,占供试品种31.2％,没有中感和高感品种。所鉴定的32个甘蓝型油菜品种中,抗病品种10个,占供试品种的31.3％,感病品种22个,占供试品种的68.8％。在芸芥中有毛类型与无毛类型有一定差异,有毛类型抗性较强。当接种菌核菌后,芸芥体内酚类物质含量迅速增加,36h后达较高水平,以后略有升高。     

鸭乙型肝炎病毒的致病性和致癌性研究──启东和高邮地区鸭的肝脏病理变化与血清中鸭乙型肝炎病毒的关系

为探讨鸭乙型肝炎病毒（ＤＨＢＶ）与鸭肝病及肝癌的关系，取鸭肝癌高发地区启东市２年以上鸭１１４只和低发地区高邮市１．５年以上鸭９８只，分别经血清学检测鸭ＤＨＢＶ，逐个进行肝脏病理组织学检查，比较两地区带毒率和阳性鸭群与阴性鸭群的病变性质及程度，采用Ｒｉｄｉｔ非参数检验方法，进行统计推断。发现无论是高邮鸭还是启东鸭，同一地区在相同条件下饲养的鸭群，ＤＨＢＶ阳性组和阴性组之间肝病的性质和程度以及肝癌检出率无显著差异；高邮阳性鸭群和启东阳性鸭群相比较，胆管增生、淀粉样变和细胞变性及肝癌检出卒等多项指标差异显著。进一步分析证明，造成这种差异的原因是由于病因不同所致。高邮鸭肝病发生的主要原因是东方次睾吸虫感染，而启东鸭肝病和肝癌发生的主要原因是黄曲霉毒素中毒。鸭乙型肝炎病毒的存在与否对两地区鸭群的肝病及肝癌的发生和发展，并无明显影响。