Effects of IL-32γ on proliferation and cell cycle of rat vascular smooth muscle cells

AIM: To observe the effects of interleukin-32γ (IL-32γ)on the proliferation and cell cycle of rat vascular smooth muscle cells (VSMCs). METHODS: The VSMCs were isolated from the thoracic aorta of SD rats by the method of tissue-piece inoculation. The cells were cultured and treated with different concentrations of IL-32γ. The proliferation of the cells was examined by MTT assay. The cell cycles were analyzed by flow cytometry. The protein levels of NF-κB p65 and cyclin D1 were detected by Western blotting. The expression of proliferating cell nuclear antigen (PCNA)was examined by immunocytochemical staining. RESULTS: Administration of IL-32γ at the concentrations of 10~50 μg/L for 24~48 h significantly promoted the proliferation of VSMCs in a dose- and time-dependent manner. After stimulation with IL-32γ at the concentration of 50 μg/L for 24 h, the cell cycle transition from G₁ phase to S/G₂ phase was accelerated and the expression levels of NF-κB p65, cyclin D1 and PCNA increased as compared with those in control group. CONCLUSION: IL-32γ promotes the proliferation of rat VSMCs and accelerates the cell cycle transition via upregulating the expression of NF-κB p65 and cyclin D1.     

Expression of IL-6 mRNA in endometrium with endometriosis

AIM: The purpose of the present study was to explore the relationship between interleukin-6 mRNA expression and endometriosis. METHODS: Using the rat model, IL-6 mRNA expression in the endometrium was examined by RT-PCR. RESULTS: The expression of IL-6 mRNA in control rats did not change at 2, 4, 6 and 8 weeks after sham operation (P>0.05), but in model rats it gradually increased at 2, 4, 6 and 8 weeks after endometriosis (P<0.01). The expression of IL-6 mRNA in uterine endometrium with endometriosis was lower than in endometriotic tissue, but higher than in endometrium from healthy controls. CONCLUSION: The IL-6 mRNA expression may contribute to the development of endometriosis . The increase in IL-6 mRNA expression may promote the implantation and growth of endometriotic tissue.     

Effects of NF-κB "decoy" oligodeoxynucleotides on TNF-α and IL-6 expression in LPS-induced mouse macrophages

AIM: To study the effect of NF-κB "decoy" oligodeoxynucleotides on TNF-α and IL-6 expression in LPS-induced mouse macrophages. METHODS: Mouse macrophage cell line J774.1 cells were cultured with LPS and liposome-mediated oligodeoxynucleotides, and the levels of TNF-α and IL-6 measured in the different culture supernatant by enzyme linked immunosorbent assay. RNA was extracted from macrophages, and the mRNA expression of TNF-α and IL-6 in macrophages was observed by RT-PCR. RESULTS: NF-κB "decoy" oligodeoxynucleotides decreased the expression of TNF-α and IL-6 in LPS-induced macrophages and inhibited generation of TNF-α and IL-6. The level of TNF-α and IL-6 did not change in control group. CONCLUSIONS: NF-κB "decoy" oligodeoxynucleotides inhibit the expression of TNF-α and IL-6 in LPS-induced macrophages, which is probably due to the specific inhibition of activated NF-κB binding sites .     

Relationship between Th17 cells infiltrated in lungs and pulmonary vascular reconstruction under hypobarric hypoxia

AIM: To investigate the changes of retinoid-related orphan receptor γt(RORγt) mRNA and interleukin-17(IL-17) protein in the lung tissue under hypobaric hypoxia, and the relationship between Th17 cells and hypoxic pulmonary vascular reconstruction. METHODS: Male BALB/c mice(n=50) were randomly divided into control group and 3 d, 7 d, 14 d and 28 d hypobaric hypoxia groups. The mice in hypobaric hypoxia groups were housed in a hypobaric hypoxia chamber(simulated altitude of 6 000 m) for 3 d, 7 d, 14 d or 28 d. The mice in control group were housed in normal pressure and oxygen environment. The hemodynamic data were recorded by cardiac catheterization. The hypertrophy of right ventricle was evaluated by the ratio of weight of the right ventricle to the weight of the left ventricle plus interventri-cular septum, and the right ventricular weight over body weight. The spleen was collected and the proportions of the Th17(CD4⁺IL-17⁺RORγt⁺) cells were detected by flow cytometry. The serum levels of IL-4, IL-6 and IL-17 and the change of IL-17 in the lung tissue were measured by ELISA. The mRNA expression of RORγt in the spleen and lung tissues was measured by RT-qPCR. RESULTS: Compared with control group, the mouse right ventricular systolic pressure, the hypertrophy index of right ventricle and the serum IL-17 level were significantly elevated in hypoxia groups, which was consistent with the results of flow cytometry. The mRNA expression of RORγt in the lung tissue was also significantly increased in 7 d, 14 d and 28 d hypoxia groups. The expression of IL-17 in the lung tissue was significantly increased in 14 d and 28 d hypoxia groups. CONCLUSION: Hypoxia promotes differentiation of Th0 cells to Th17 cells in the spleen. The Th17 cells infiltrated in the lung tissue under hypobarric hypoxia are involved in pulmonary vascular reconstruction.     

Effects of microglia-derived IL-1β on differentiation of OPCs in corpus callosum of septic neonatal rats

AIM: To explore whether IL-1β inhibits the oligodendrocyte precursor cell (OPCs) differentiation and affects axonal myelination. METHODS: One-day-old SD rats were randomly divided into control group and LPS group (48 rats in each group). The rats in LPS group were intraperitoneally injected with 1 mg/kg LPS. The rats in control group were injected with an equal volume of PBS. The rats in each group were further divided into 3 h, 24 h, 3 d, 7 d, 14 d and 28 d subgroups after injection. The expression of IL-1β and IL-1R₁ in the rat corpus callosum at 3 h, 24 h, 3 d, 7 d was determined by double immunofluorescence and Western blotting. The myelin basic protein(MBP) expression in the rat corpus callosum at 14 d, 28 d after injection was also measured. In vitro, primary OPCs culture was performed and divided into control group, 30 μg/L IL-1β group, 30 μg/L IL-1β+IL-1Ra group and 30 μg/L IL-1Ra group. The expression of MBP in the OPCs induced differentiation for 3 d was observed by double immunofluorescence and Western blotting. RESULTS: The expression of IL-1β and IL-1R₁ in the rat corpus callosum at 3 h, 24 h, 3 d, 7 d after LPS injection was obviously increased and the expression of MBP in the rat corpus callosum at 14 d, 28 d in LPS group was obviously decreased compared with control group in vivo. The level of MBP was significantly decreased after IL-1β treatment for 3 d in vitro. However, IL-1Ra (IL-1R inhibitor) reversed the down-regulation of MBP expression. IL-1β inhibited the expression of p-ERK, ERK over-expression reversed the down-regulation of MBP expression compared with IL-1β group. CONCLUSION: IL-1β inhibits the differentiation of OPCs, which may be involved in ERK pathways, thus leading to axonal hypomyelination in the corpus callosum of septic neonatal rats.     

NDV F与ChIL-18融合基因"自杀性"质粒的构建

根据GeneBank公布的鸡新城疫病毒(NDV)Lasota株F基因和鸡源性白细胞介素18(ChIL-18)基因序列分别设计特异性引物,且在鸡NDV F基因引物上游引入BamH Ⅰ位点、下游引入Sal Ⅰ位点,在Ch IL-18基因引物上游引入Sal Ⅰ位点、下游引入Hind Ⅲ、BamH Ⅰ位点.以鸡新城疫病毒Lasota株RNA为模板RT-PCR扩增其F基因,将F基因插入pMD18-T的BamH Ⅰ与Sal Ⅰ位点中.同时扩增上游带有Sal Ⅰ位点、下游带有Hind Ⅲ、BamH Ⅰ位点的ChIL-18基因,并将其插入到含有F基因的pMD18-T质粒的Sal Ⅰ、Hind Ⅲ位点之间,从而使NDV F基因与ChIL-18基因形成具有一个完整阅读框的融合基因,然后将融合基因片段定向克隆到pSFV单一启动子(CMV)下游,构建成F-ChIL-18融合基因重组"自杀性"质粒pSFV-F-ChIL-18.     

子莲新品种——“建选17号”亲本分子鉴定

建选17号系红花建莲/寸三莲65//太空莲2号复合杂交选育而成的子莲新品种。利用RAPD和ISSR两种分子标记技术,对4个莲子基因型的亲缘关系进行鉴定。结果显示,莲子亲本之间具有丰富的遗传背景;在所检测到的多态性位点中,建选17号与红花建莲相同的有14个,占25.93%;与寸三莲65相同的有11个,占20.37%;与太空莲2号相同的有32个,占59.26%。但是ISSR检测到1个位点(ISSR-L8-950bp)为建选17号所特有。建选17号97.44%的遗传信息分别来自不同的亲本基因型,并发生了部分遗传变异;与太空莲2号、寸三莲65、红花建莲的遗传距离分别为0.1467、0.5792、0.6646。从而从DNA水平证实了建选17号确系3个亲本的复合杂交后代。     

不同浓度雌二醇对猪卵母细胞体外成熟和孤雌激活胚胎的影响

为了探讨雌二醇（17β-estrodiol,E2）对猪卵母细胞体外成熟及孤雌激活后胚胎早期发育的影响,在卵母细胞体外成熟培养基中添加不同浓度雌二醇,研究卵裂率和囊胚率的变化。以未添加雌二醇的基础液为对照组,比较分析各组卵母细胞核成熟效率、孤雌激活后胚胎的卵裂率、囊胚发育率。结果表明,成熟液中添加1μg／mL雌二醇（E2）对猪卵母细胞的体外成熟具有明显的促进作用,而添加100μg／mL雌二醇（E2）对猪卵母细胞体外成熟具有明显的抑制作用。     

长期干旱胁迫下Mdm-miR160- MdARF17- MdHYL1模块调控苹果生长发育的研究

为了研究长期干旱条件下Mdm-miR160-MdARF17-MdHYL1 调控网络在苹果生长发育中的作用,以GL-3和Mdm-miR160e OE、MdARF17 RNAi、MdmARF17 OE、 MdHYL1 OE及MdHYL 1 RNAi转基因苹果为试验材料,进行为期3个月的长期干旱处理,测定苹果株高、根干质量、根冠比以及叶片净光合速率、胞间CO₂浓度、气孔导度、蒸腾速率、叶绿素荧光值和水分利用效率。结果表明,在长期干旱胁迫下, Mdm-miR160e OE转基因苹果植株更矮化,根冠比、根干质量、光合能力及水分利用效率大于GL-3植株,具有更强的耐旱性; MdARF17 RNAi植株比GL-3更矮,且具有更大的根冠比、根干质量、光合能力和水分利用效率,而MdARF17 OE与之相反; MdHYL1 RNAi植株对长期干旱胁迫更敏感,其株高大于野生型且具有更小的根冠比、更低的光合能力和水分利用效率,而 MdHYL1 OE植物对长期干旱条件具有更强的适应性,生长发育表型与RNAi植株相反。综上所述,Mdm-miR160- MdARF17-MdHYL1调控网络在苹果响应长期干旱过程中发挥了重要功能,为苹果抗旱分子育种提供了候选基因。     

Role of type II innate lymphoid cells in browning of white adipose tissue

Type II innate lymphoid cells (ILC2s) are widely distributed in the blood, intestines, trachea, lung, spleen, liver, animal fat and skin, and involved in the innate immune responses. ILC2s have attracted much attention for its important roles in the conversion of white adipose to beige adipose. Studies have shown that ILC2s are essential for the proliferation and differentiation of adipocyte precursor cells, and they also play a vital role in anti-parasitic infection and allergic inflammation. This review discusses the discovery, differentiation, development, distribution and function of ILC2s, and their relationships with the browning of white adipose tissue for providing valuable references on understanding the pathogenesis, prevention and treatment of obesity and fat metabolism disorders.