Increased intratumoral IL-22-producing CD4+ T cells and Th22 cells correlate with gastric cancer progression and predict poor patient survival

IL-22-producing CD4⁺ T cells (IL-22⁺CD4⁺ T cells) and Th22 cells (IL-22⁺IL-17^?IFN-γ^?CD4⁺ T cells) represent newly discovered T-cell subsets, but their nature, regulation, and clinical relevance in gastric cancer (GC) are presently unknown. In our study, the frequency of IL-22⁺CD4⁺ T cells in tumor tissues from 76 GC patients was significantly higher than that in tumor-draining lymph nodes, non-tumor, and peritumoral tissues. Most intratumoral IL-22⁺CD4⁺ T cells co-expressed IL-17 and IFN-γ and showed a memory phenotype. Locally enriched IL-22⁺CD4⁺ T cells positively correlated with increased CD14⁺ monocytes and IL-6 and IL-23 detection ex vivo, and in vitro IL-6 and IL-23 induced the polarization of IL-22⁺CD4⁺ T cells in a dose-dependent manner and the polarized IL-22⁺CD4⁺ T cells co-expressed of IL-17 and IFN-γ. Moreover, IL-22⁺CD4⁺ T-cell subsets (IL-22⁺IL-17⁺CD4⁺, IL-22⁺IL-17^?CD4⁺, IL-22⁺IFN-γ⁺CD4⁺, IL-22⁺IFN-γ^?CD4⁺, and IL-22⁺IL-17⁺IFN-γ⁺CD4⁺ T cells), and Th22 cells were also increased in tumors. Furthermore, higher intratumoral IL-22⁺CD4⁺ T-cell percentage and Th22-cell percentage were found in patients with tumor-node-metastasis stage advanced and predicted reduced overall survival. In conclusion, our data indicate that IL-22⁺CD4⁺ T cells and Th22 cells are likely important in establishing the tumor microenvironment for GC; increased intratumoral IL-22⁺CD4⁺ T cells and Th22 cells are associated with tumor progression and predict poorer patient survival, suggesting that tumor-infiltrating IL-22⁺CD4⁺ T cells and Th22 cells may be suitable therapeutic targets in patients with GC.     

Structure,composition and mechanical properties of the silk fibres of the egg case of the Joro spider, <Emphasis Type="Italic">Nephila clavata</Emphasis> (Araneae,Nephilidae)

The silk egg case and orb web of spiders are elaborate structures that are assembled from a number of components. We analysed the structure, the amino acid and fibre compositions, and the tensile properties of the silk fibres of the egg case of Nephila clavata. SEM shows that the outer and inner covers of the egg case consist of thick, medium and thin silk fibres. The silk fibres of the outer cover of the egg case are probably produced by the major and minor ampullate glands. The silk fibres of the inner cover of the egg case from cylindrical glands appears to be distinct from the silk fibres of the major ampullate glands based on their micro-morphology, mole percent amino acid composition and types, and tensile behaviour and properties. Collectively, our investigations show that N. clavata uses silk fibres from relatively few glands in varying combinations to achieve different physical and chemical properties (e.g., color, diameter, morphology and amino acid composition) and functional and mechanical properties in the different layers of the egg case.     

Over-expression of human carcinoma-associated antigen in intrahepatic cholangiocellular carcinoma

Objective: To investigate the expression status of human carcinoma antigen (HCA) in human cholangiocellular carcinomas, and to determine the relationship between HCA and clinical features. Methods: Tissues from 60 intrahepatic cholangiocellular carcinoma (ICC) patients, and normal liver tissues from 20 hepatic hemangioma patients selected randomly were assayed for the expression of HCA by immunohistochemistry, and Western blots. Areas of poorly differentiated (n = 20), moderately-well differentiated (n = 30), highly differentiated tumors (n = 10) from different cases were evaluated. Results were recorded as positive (?5% of cells staining and staining intensity 2+ or 3+) or negative (<5% of cells staining and staining intensity <2+) and analyzed using the χ² test. Results: BCE075 and BDD048 antibodies showed similar staining patterns. The positive immunostaining of BCE075 was mainly localized in the cytoplasm and cell secretions. The staining was positive in 15% of poorly differentiated ICC, 72% of moderately-well differentiated, 100% of highly differentiated tumors. But, staining was not detected in adjacent normal tissue. The differences in HCA expression among these tissues were statistically significant. Also, we found expression of HCA to be closely associated with the degree of differentiation of ICC and tumor cell morphology. There was a correlation between expression of HCA and serum CA19-9. Conclusion: The data suggest that HCA is a potential marker for the diagnosis of cholangiocellular carcinoma.     

Diet composition of post-metamorphic bullfrogs (Rana catesbeiana) in the Zhoushan archipelago, Zhejiang Province, China

Bullfrogs (Rana catesbeiana) are listed as one of the 100 worst invasive alien species in the world. They are generalist predators and thus may affect native species through predation. In previous studies, the food contents of bullfrogs were mostly examined at a single site. In the present study, the diet composition of post-metamorphic bullfrogs on eight islands (Daishan, Liuheng, Xiushan, Fodu, Taohua, Xiashi, Cezi, and Putuoshan) in the Zhoushan Archipelago, Zhejiang Province of China, were examined by using the stomach flushing method from June 30 to August 11 in 2005. A total of 391 individual frogs were measured, including 113 adults and 278 juveniles. The analysis of the stomach contents shows that, for adult bullfrogs, the most important prey (by diet volume) overall were Decapoda, Coleoptera, Odonata, Mesogastropoda, Raniformes, and Cypriniformes. For juvenile bullfrogs, these were Decapoda, Coleoptera, Cypriniformes, Odonata, Orthoptera, Hymenoptera, Lepidoptera larvae, Mesogastropoda, and Raniformes. Moreover, the prey size and diet volume increased with the body size of both adult and juvenile bullfrogs. The diet composition of primary preys of bullfrogs was significantly different among the islands. The results indicate that bullfrogs exert different predatory influences on native fauna at different sites and that bullfrogs are generalist predators with extensive ecological impacts on native fauna. __________ Translated from Biodiversity Science, 2006, 14 (5): 363–371 [译自: 生 物多样性]     

蒙古栎叶片及其土壤碳、氮同位素自然丰度对大气CO2浓度升高的响应

大气CO₂浓度升高对土壤氮素转化过程产生重要影响,研究其变化有助于更好地预测陆地生态系统的固碳潜力.氮同位素自然丰度作为生态系统氮素循环过程的综合指标能够有效地指示CO₂浓度升高对土壤氮素转化过程的影响.本研究采用开顶箱CO₂ 熏蒸法研究连续10年的大气CO₂ 浓度升高对我国东北地区蒙古栎及其土壤和微生物生物量碳、氮同位素自然丰度的影响.结果表明: 大气CO₂浓度升高改变了土壤氮循环过程,增加了土壤微生物和植物叶片δ¹⁵N;促进了富¹³C土壤有机碳分解,中和了贫¹³C植物光合碳输入的效果,导致土壤可溶性有机碳和微生物碳δ¹³C在CO₂升高条件下没有发生显著变化.这些结果表明,CO₂浓度升高很可能促进了土壤有机质矿化过程,并加剧了系统氮限制的状态.     

An N-terminal amphipathic helix in hepatitis C virus (HCV) NS4B mediates membrane association, correct localization of replication complex proteins, and HCV RNA replication

Like other positive-strand RNA viruses, hepatitis C virus (HCV) is believed to replicate its RNA in association with host cell cytoplasmic membranes. Because of its association with such membranes, NS4B, one of the virus's nonstructural proteins, may play an important role in this process, although the mechanistic details are not well understood. We identified a putative N-terminal amphipathic helix (AH) in NS4B that mediates membrane association. Introduction of site-directed mutations designed to disrupt the hydrophobic face of the AH abolishes the AH's ability to mediate membrane association. An AH in NS4B is conserved across HCV isolates. Completely disrupting the amphipathic nature of NS4B's N-terminal helix abolished HCV RNA replication, whereas partial disruption resulted in an intermediate level of replication. Finally, immunofluorescence studies revealed that HCV replication complex components were mislocalized in the AH-disrupted mutant. These results identify a key membrane-targeting domain which can form the basis for developing novel antiviral strategies.     

稀有鮈鲫眼的形态发生研究

详细观察和描述了稀有鮈鲫眼发育的形态变化,并分别对视网膜发育早期神经层、色素上皮层的厚度及眼发育后期视网膜的总厚度进行了测量。结果表明:稀有鲫眼的发生始于神经胚时期形成的眼原基,与两栖、哺乳类动物不同,其眼原基是由间脑向左右伸出的对称外突实体;在尾芽期眼原基向腹侧弯曲、侧向伸长,随后开始内陷,在尾泡期形成视杯。眼原基外层与外胚层紧贴的部分将发育为视杯的内层、神经视网膜,而眼原基其余部分将分化为视杯的外层、视网膜色素层。在尾泡期之后,视网膜色素层停止有丝分裂开始形成色素颗粒,此时视网膜神经层开始分化。视网膜这两层结构在发育早期分化的有序进行可能与早期胚胎头部内空间以及附近的间充质细胞有关。从神经胚期到尾泡期,视网膜色素层厚度由42.3±0.8μm减小到4.8±0.4μm,视网膜神经层厚度从37.1±0.2μm增加到43.7±0.6μm,而从尾鳍期到孵出期视网膜总厚度从42.7±1.2μm逐渐增加到98.3±2.1μm。与所有脊椎动物一样,稀有鲫眼的视网膜分化也是按照由内向外的顺序进行,晶状体、角膜及其他结构在孵出时已基本发育完全。     

Invariant chain modulates HLA class II protein recycling and peptide presentation in nonprofessional antigen presenting cells

The expression of MHC class II molecules and the invariant chain (Ii) chaperone, is coordinately regulated in professional antigen presenting cells (APC). Ii facilitates class II subunit folding as well as transit and retention in mature endosomal compartments rich in antigenic peptides in these APC. Yet, in nonprofessional APC such as tumors, fibroblasts and endocrine tissues, the expression of class II subunits and Ii may be uncoupled. Studies of nonprofessional APC indicate class II molecules access antigenic peptides by distinct, but poorly defined pathways in the absence of Ii. Here, investigations demonstrate that nonprofessional APC such as human fibroblasts lacking Ii internalize antigenic peptides prior to the binding of these ligands to recycling class II molecules. By contrast, fibroblast lines expressing Ii favor exogenous peptides binding directly to cell surface class II molecules without a need for ligand internalization. Endocytosis of class II molecules was enhanced in cells lacking Ii compared with Ii-expressing APC. These results suggest enhanced reliance on the endocytic recycling pathway for functional class II presentation in nonprofessional APC.     

Pre-treatment of recombinant mouse MFG-E8 downregulates LPS-induced TNF-α production in macrophages via STAT3-mediated SOCS3 activation

Milk fat globule-epidermal growth factor factor 8 (MFG-E8) regulates innate immune function by modulating cellular signaling, which is less understood. Herein, we aimed to investigate the direct anti-inflammatory role of MFG-E8 in macrophages by pre-treatment with recombinant murine MFG-E8 (rmMFG-E8) followed by stimulation with LPS in RAW264.7 cells and in peritoneal macrophages, isolated from wild-type (WT) or MFG-E8(-/-) mice. RAW264.7 cells and mouse peritoneal macrophages treated with rmMFG-E8 significantly downregulated LPS-induced TNF-α mRNA by 25% and 24%, and protein levels by 29% and 23%, respectively (P<0.05). Conversely, peritoneal macrophages isolated from MFG-E8(-/-) mice produced 28% higher levels of TNF-α, as compared to WT mice when treated with LPS. In in vivo, endotoxemia induced by intraperitoneal injection of LPS (5 mg/kg BW), at 4 h after induction, serum level of TNF-α was significantly higher in MFG-E8(-/-) mice (837 pg/mL) than that of WT (570 pg/mL, P<0.05). To elucidate the direct anti-inflammatory effect of MFG-E8, we examined STAT3 and its target gene, SOCS3. Treatment with rmMGF-E8 significantly induced pSTAT3 and SOCS3 in macrophages. Similar results were observed in in vivo treatment of rmMFG-E8 in peritoneal cells and splenic tissues. Pre-treatment with rmMFG-E8 significantly reduced LPS-induced NF-κB p65 contents. These data clearly indicated that rmMFG-E8 upregulated SOCS3 which in turn interacted with NF-κB p65, facilitating negative regulation of TLR4 signaling for LPS-induced TNF-α production. Our findings strongly suggest that MFG-E8 is a direct anti-inflammatory molecule, and that it could be developed as a therapy in attenuating inflammation and tissue injury.