FcεRIα is responsible for high-affinity binding with the Fc portion of IgE, which is critical for IgE-dependent disease responses such as allergy responses and anti-parasite immunity. FcεRIα is expressed on a few cell types, including mast cells and basophils.

Functioned as α-subunit of the high-affinity immunoglobulin E receptor (Fcε RIα), Fcε RIα plays a central role in the pathogenesis of Ig-E-mediated allergy and other IgE-related disorders. Fcε RIα is normally expressed only in limited spectrum of cells like basophils and mast cells, but the mechanism of controlling Fcε RIα expression in these cells is less well understood. In this study, we found fully overlapped natural antisense transcript (NATs) of Fcε RIα ( FCER1A -AS) is co-expressed with cognate sense transcript ( FCER1A -S) in IL-3 induced Fcε RIα-expressing cells or in high Fcε RIα-expressing cell line MC/9. When FCER1A -AS is selectively knocked down by CRISPR/RfxCas13d (CasRx) approach in MC/9, expression of mRNA and proteins of FCER1A -S is also markedly decreased. Furthermore, deficiency of FCER1A -AS along with lack of FCER1A -S expression is found in two different lines of gene-targeted mice in which FCER1A locus is disturbed at different sites. More importantly, the FCER1A -AS-deficient homozygous mice display similarly diminished anaphylactic reaction as FCER1A gene knockout mice. Thus, we uncovered in this investigation that the expression of FCER1A -S is positively regulated by co-expressed fully overlapped antisense transcript.

综述了近年来我国水环境中四环素类抗生素的污染现状及其对水生生物的毒性效应的研究进展.总体而言,四环素类抗生素在我国的生产和使用量较大且缺乏相应的排放标准,导致四环素类抗生素在我国水环境污染情况严峻;高浓度的四环素类抗生素急性暴露对水生生物具有一定的致死和亚致死效应,在环境浓度长期暴露下会影响水生生物的正常生长.目前对四环素类抗生素生态毒性的研究还有一些不足之处,例如,对四环素类抗生素与其它污染物的联合暴露及其降解产物的毒性的了解有限,尚需更多研究以全面地评价四环素类抗生素在环境中的风险.

目的 探究细胞因子IL-4抑制骨髓细胞分化产生嗜碱性粒细胞的能力及机制.方法 采用IL-3诱导培养小鼠骨髓细胞,流式细胞术检测FcεRIα+ckit-CD49b+嗜碱性粒细胞以及FcεRIα+ckit-CD49b-新发现的单阳细胞群,并用Ki67检测其增殖状态.流式细胞术分选单阳细胞群后继续用IL-3诱导培养,以确定其后续分化能力.结果 经IL-3诱导培养的骨髓细胞,3～4d后开始有嗜碱性粒细胞出现,至第9天细胞比例达到高峰,然后开始下降.与野生型相比,IL-4缺陷的骨髓细胞诱导出的嗜碱性粒细胞,无论是在细胞比例或数目还是该细胞表达FcεRIα的平均荧光强度都显著增加.在IL-4缺陷型骨髓细胞诱导过程中加入外源重组IL-4可抑制嗜碱性粒细胞的产生;而在野生型骨髓细胞诱导过程中加入抗IL-4抗体则可增加嗜碱性粒细胞的产生,提示诱导过程中产生的IL-4具有负向调控嗜碱性粒细胞形成的作用.与野生型相比,IL-4缺陷鼠骨髓细胞经诱导可产生更多未曾报道过的表型为FcεRIα+ckit-CD49b-的单阳细胞群,且该细胞群的Ki67阳性率更高.单阳细胞群经IL-3刺激后可继续分化出嗜碱性粒细胞.结论 IL-3诱导培养骨髓细胞过程中,内源产生的IL-4可抑制嗜碱性粒细胞的产生及FcεRIα 的表达水平.

The effects of microplastics (MPs) on organisms have drawn a worldwide attention in the recent years. In this study, zebrafish embryos were employed to assess the combined effects of MPs and cadmium (Cd) on the aquatic organisms. Lethal and sublethal effects were recorded at 8, 24, 32, 48 and 96 hpe (hour post exposure, hpe). The exposure under a series concentration of MPs and/or an environmental level Cd has the negative impacts on survival and heart rate (HR). And there was a positive correlation between MPs concentration and lethal and sublethal toxicity under combined exposure. The physiological parameters showed that the mixture of two stressors had the antagonistic toxicity under low concentration of MPs (0.05, 0.1 mg/L) while the synergistic sublethal toxicity under high levels of MPs (1, 5, 10 mg/L) on zebrafish embryos. Both the scanning electron micrographs (SEM) and fluorescence microscope photos suggested an electrostatic interaction and weak physical forces generated between MPs and chorion membrane. It is inferred that the 10 μm MPs could induce the protective effect of chorion membrane and cause complex toxicities with Cd. But when it involved with other pollutants, the toxic effects and mechanism are still waiting to be figured out.

As a primary degradation by-product of tetracycline (TC), 4-Epianhydrotetracycline (4-EATC) has been detected frequently in the aquatic environment, which may pose a potential environmental risk to aquatic organisms. Up to now, however, the toxicology study on 4-EATC to aquatic organisms is limited. In the present study, in order to better understand the toxic mechanism of 4-EATC, developmental toxicity including lethal and sublethal effects of 4-EATC and TC were investigated. The results showed that the developmental toxicity of 4-EATC to zebrafish embryos was stronger than that of TC. The 96 h LC50 value of 4-EATC to zebrafish embryos was 29.13 mg/L. Malformations seemed to be the most sensitive sublethal endpoint of 4-EATC exposure, and the 96 h EC50 value was 8.57 mg/L. Transcriptome response of 4-EATC to zebrafish embryos was determined. The results showed that 430 different expression genes (DEGs) caused by 4-EATC, and most enriched in tryptophan (TRP) metabolism pathway. Annotation of DEGs in the TRP metabolism demonstrated that expression of 4 gene products in tryptophan metabolized along the kynurenine (KYN) pathway were changed. Disorder of TRP catabolism in KYN pathway was a potential mechanism of 4-EATC toxicity to zebrafish embryos.

Background Sediments function as a secondary and significant source of tributyltin (TBT) and triphenyltin (TPT) in aquatic ecosystems and may pose a potential threat on benthic organisms and human health. The subchronic toxicity of sediment-associated TBT or TPT to snailsBellamya aeruginosaat environmentally relevant concentrations was investigated in this study. Multiple endpoints at the biochemical [ethoxyresorufin-O-deethylase (EROD), superoxide dismutase (SOD), catalase (CAT), protein carbonyl content (PCC) and lipid peroxidation (LPO)] and transcriptomic levels were examined. Results TBT or TPT in sediment could induce antioxidant enzymes' activities and result in oxidative damage in the hepatopancreas ofB. aeruginosaafter 28-day exposure. A transcriptomic profile ofB. aeruginosaexposed to TBT and TPT was reported. CYP genes and EROD activity were sensible and reliable biomarkers for toxicity assessment of TBT or TPT in sediments. Comparative pathway analysis revealed the alteration of steroid hormone biosynthesis and retinol metabolism inB. aeruginosaafter 90-day exposure to sediment-associated TBT at the concentration of 2000 ng/g dw, which might affect both reproduction and lipogenesis functions. The ubiquitin proteasome system and immune system might be the toxicity target inB. aeruginosaafter exposure to sediment-associated TPT for 90 days. Conclusions The results offered new mechanisms underlying the toxicity of sediment-associated tributyltin and triphenyltin.

Guided by the educational concept of outcomes based education ( OBE), based on the bridge professional knowledge of pathology, medical microbiology, medical immunology as well as the pathogenesis and outcome of diseases in almost all organs and systems of human body in the teaching of basic medicine courses, aiming at their characteristics of response between knowledge points, broad contents and close connection with clinical practice, an English integrated teaching module of "Injury, Infection, Immunity and Repair" in basic medicine was constructed. The modular teaching proceeds from knowledge structure to reverse curriculum design, implements student-centered and diversified teaching and learning methods, and continuously improves teaching evaluation and guarantee system. It optimizes the shortcomings of traditional subject teaching system and organ system integration curriculum system, further improves the quality of medical education, promotes the construction of basic medicine, advances the globalization process of medical education, and cultivates more medical professionals for our country to adapt to the global development in the future.

Aluminium hydroxide (alum), the most widely used adjuvant in human and animal vaccines, has long been known to promote T helper type 2 (Th2) responses and Th2-associated humoral responses, but the mechanisms have remained poorly understood. In this study, we explored whether alum is able to directly modulate antigen-presenting cells to enhance their potency for Th2 polarization. We found that alum treatment of dendritic cells failed to show any Th2-promoting activities. In contrast, alum was able to enhance the capacity of basophils to induce Th2 cells. When basophils from interleukin-4 (IL-4) knockout mice were examined, the intrinsic Th2-promoting activities by basophils were largely abrogated, but the alum-enhanced Th2-promoting activities on basophils were still detectable. More importantly, Th2-promoting adjuvant activities by alum found in IL-4 knockout mice were also largely reduced when basophils were depleted by antibody administration. Therefore, basophils can mediate Th2-promoting activities by alum both in vitro and in vivo through IL-4-independent mechanisms. Further studies revealed that secreted soluble molecules from alum-treated basophils were able to confer the Th2-promoting activities, and neutralization of thymic stromal lymphopoietin or IL-25 attenuated the IL-4-independent development of Th2 cells elicited by alum-treated basophils. Finally, alum was able to activate NACHT, LRR and PYD domains-containing protein 3 (NLRP3) inflammasome in murine basophils in the same way as alum in professional antigen-presenting cells, but NLRP3 was not required for Th2-promoting activities on basophils by alum in vitro. These results demonstrated that alum can enhance the capacities of basophils to polarize Th2 cells via IL-4- and NLRP3-independent pathways.

This article has been accepted for publication and undergone full peer review but has not been through the copyediting, typesetting, pagination and proofreading process which may lead to differences between this version and the Version of Record. Please cite this article as an 'Accepted Article', doi: 10.1111/imm.12636 This article is protected by copyright. All rights reserved. Received Date : 31-Oct-2015 Revised Date : 11-Jun-2016 Accepted Date : 12-Jun-2016 Article type : Original Article

Although dendritic cells (DCs) have been widely demonstrated to play essential roles in initiation of Th2 responses in helminth infections and allergic reactions, the mechanisms remain uncertain largely because DCs do not produce IL-4. In present investigation, we have uncovered a novel subset of DCs from mice infected with Th2-provoking pathogens Schistosoma japonica, which independently promoted Th2 cells via IL-4–dependent pathway. These DCs contained similar levels of IL-4 mRNA and higher levels of IL-12p40 mRNA comparing to basophils, correlating to their Th2-promoting and Th1-promoting dual polarization capacities. Characterized by expression of FcεRI+, these DCs were induced independent of T cells. Further investigations revealed that Th2-promoting FcεRI+ DCs were monocyte-derived inflammatory DCs, which were sufficient to induce Th2 cells in vivo. Egg Ags together with GM-CSF or IL-3 alone were able to stimulate the generation of Th2-promoting FcεRI+ DCs from bone marrow cells in vitro. To our knowledge, our data for the first time demonstrate that IL-4–producing DCs are induced under some Th2-provoking situations, and they should play important roles in initiation of Th2 response.

BACKGROUND:It has been well accepted that glycans present in schistosomes are highly antigenic. However, it is not clear what kind of worm glycans can affect the infected host to mount IgG responses and whether mounted anti-glycan IgG responses are protective.METHODS:The contribution of antigenicity by glycans was measured by using competitive ELISA assay in sera from infected mice and humans. Monoclonal antibodies towards soluble Schistosoma japonicum egg antigens (SjEA) were generated from SjEA immunizated mice. The expression of glycans on surfaces of cercaria or young worm and their distributions were examined by immunofluorescence assay. The protective roles of glycans-specific mAbs were assayed by determination of the worm and egg burden in infected mice.RESULTS:Both periodate-resistant glycans and periodate-sensitive glycans are antigenic in schistosome infections. When monoclonal antibodies against either periodate-sensitive or periodate-resistant glycans were administered prior to schistosome infections in mice, both kinds of anti-glycan antibodies were found to successfully provide protective immunity to infected mice.CONCLUSIONS:Both periodate-resistant and periodate-sensitive glycans are antigenic, and dominant anti-glycan IgG responses can play important roles in protective immunity in schistosome infected hosts.

目的 筛选K48聚泛素链在肿瘤坏死因子受体相关因子2(tumor necrosis factor receptor-associated factor 2,TRAF2)上的主要修饰位点.方法 比较不同物种间的TRAF2氨基酸序列,选出人TRAF2上保守率在90％以上的赖氨酸.构建人野生型TRAF2的表达载体及保守赖氨酸突变为精氨酸的突变表达载体.将不同的TRAF2表达载体与NF-κB荧光素酶报告基因表达载体共转至293FT细胞中,通过荧光素酶检测NF-κB激活情况.结果 人TRAF2上共有8个保守率在90％以上的赖氨酸,酶切及测序结果证明本研究成功构建TRAF2野生型和突变型表达载体.NF-κB荧光素酶报告基因检测证明TRAF2-K320可能是K48聚泛素链的主要修饰位点.结论 TRAF2-K320位点对TRAF2介导的NF-κB激活具有负向调控作用.

OBJECTIVE:To investigate the effect of calpeptin on diaphragmatic injury and atrophy under controlled mechanical ventilation in rats.METHODS:A total of 24 SPF Sprague-Dawley (SD) rats were randomly divided into anesthetized control group (CON group), 24-hour controlled mechanical ventilation group (CMV group), and 24-hour CMV + treatment with calpeptin group (CMVC group), with 8 rats in each group. Animals in the CON group received an intraperitoneal injection of pentobarbital sodium without CMV and continuous infusion of pentobarbital sodium. A small-animal ventilator was used for 24 hours in rats of CMV group. Rats of CMVC were treated with a specific calpain inhibitor calpeptin (4 mg/kg). The drug was injected subcutaneously 2 hours before and 8, 15 and 23 hours after mechanical ventilation. Changes in diaphragm ultrastructure, light microscopic picture, and myosin heavy chain (MHC) expression were observed.RESULTS:(1) Alignment of myofilaments and normal Z-band, and the shape of mitochondria were maintained in CON group as revealed by electron microscope. The signs of misalignment of myofibrils, disruption of Z-band and vacuolar mitochondria were found in CMV group, and they were obviously improved in CMVC group. The density of muscle injury (× 10⁻²/μm²) in CMV group was significantly higher than that in control group (36.8 ± 13.7 vs. 6.4 ± 6.3, t=6.373, P=0.001), and that in CMVC group was significantly lowered (17.6 ± 9.1 vs. 36.8 ± 13.7, t=3.694, P=0.002).(2) In CON group, the diaphragm fibers appeared regular in cross section without pathologic change under light microscopy. Fuzzy muscle striations, irregular muscle fibers, centralized nuclei and swelling of capillary endothelial cells were observed in CMV group, while pathological changes in the CMVC group were milder significantly. (3) In CMV group, the density of MHCslow and MHCfast was lower compared with that of CON group, and the gray value was lowered by 61.1% (t=8.138, P=0.001) and 77.1% (t=8.844, P=0.001), respectively, especially in MHCfast. However, the gray values of MHCslow and MHCfast were increased by 1.51 folds (t=4.601, P=0.010), and 1.33 folds (t=2.859, P=0.011), respectively, after treatment with calpeptin, and the elevation was more significantly in MHCslow.CONCLUSIONS:Diaphragmatic injury and atrophy were found after CMV for 24 hours. Calpeptin could reverse the detrimental effects of CMV, and it suggested that calpain plays an important role in modulating the ventilator-induced dysfunction of the diaphragm.

BACKGROUND:Schistosomiasis is a chronic infection, where the host immune response to the parasite changes from a predominantly Th1 to Th2 phenotype, when parasite enters the egg stage, restraining the host inflammatory immune responses to achieve a longer survival in the host. On the other hand, the development of Th2 responses causes immunopathological changes such as liver fibrosis. Therefore identification of schistosome-derived Th2 inducing molecules is important in the understanding of pathogenesis of schistosomiasis. A cyclophilin A homologue of Schistosoma japonicum was reported to be an egg-stage specific antigen, but its immunogenicity and immunoregulatory activities remain unknown.METHODS:We cloned and expressed the gene of cyclophilin A from Schistosoma japonicum (AY814078), named as SjCyP18 based on its molecular weight. The expression profiles in different stages of S. japonicum were examined by RT-PCR and immunofluorescence assay. The immunogenicity of SjCyP18 was measured by the presence of IgG in the sera from S. japonicum infected patients and animals, and the Th2-promting activities were examined by the subclass of immunoglobulins against SjCyP18 and by the IL-4 induction in T cells following footpad injection of SjCyP18.RESULTS:The cloned SjCyP18 has 65% homology with human or mouse cyclophilin A at the amino acid level. In contrast to reports as an egg-stage specific antigen, the gene was found to be expressed in all stages of S. japonicum. IgG responses against SjCyP18 were found in some S. japonicum infected patients and were significantly induced when infection become patent and produce eggs in infected mice. Furthermore, the Th2-promoting subclass of IgG1 was the predominant isotype in S. japonicum infected mice. More importantly, footpad injection of SjCyP18 induced a greater production of IL-4 than that of IFN-γ by lymphocytes compared to responses from PBS injection controls.CONCLUSION:The cyclophilin A homologue found in S. japonicum is immunogenic and promotes Th2 responses in vivo which may contribute to the establishment of chronic infection by schistosomes.

UNLABELLEDOBJECTIVE; To observe the changes in various dendritic cell (DC) subsets, macrophages, basophils, eosinophils and mast cells in mouse spleen before and after Schistosoma japonicum infection induced Th2 response.METHODSC57BL6 mice were infected with 20 S. japonicum cercariae via abdominal skin. Before infection and at 2, 4, and 6 weeks post-infection, the mice were sacrificed and spleen was removed. The frequencies of non-T, non-B basophils (NTNB), eosinophils, mast cells, subsets of DC and macrophages in the spleen were measured by flow cytometry.RESULTSAt 4 weeks after infection, when Th2 cells started to occur, the proportion of CD11c+CD8+DC and CD11c+CD4+DC in B220CD11c+DC increased from 7.4% and 7.9% before infection to 17.1% and 12.0%, respectively. During the infection, CD11c+CD4-CD8-DC, the majority of B220CD11c+DC, remained on a nearly constant level (70%); the percentage of B220+ CD11c+ DC in NTNB decreased. The macrophages were subdivided into two subsets: F4/80+CD11b(int) and F4/80+CD11b(high). The percentage of F4/80+CD11b(int) and F4/80+CD11b(high) in NTNB dropped from 15.4% and 13.7% before infection to 2.7% and 8.6% at 4 weeks post-infection. The proportion of CD11b high macrophages in F4/80+ cells significantly increased from 47.1% before infection to 75.5% at 4 weeks after infection. During S. japonicum infection, eosinophil percentage in the spleen gradually increased, while the frequency of basophils and mast cells in NTNB greatly decreased.CONCLUSIONAt the time when Th2 response starts to occur, the frequency of CD11c+CD8+DC and CD11c+CD4+DC in CD11c+ dendritic cells increases. Once Th2 immune response established, the eosinophil frequency increases.

C-type lectin receptors (CLRs) play critical roles as pattern-recognition receptors (PRRs) for sensing Candida albicans infection, which can be life-threatening for immunocompromised individuals. Here we have shown that Dectin-3 (also called CLECSF8, MCL, or Clec4d), a previously uncharacterized CLR, recognized α-mannans on the surfaces of C. albicans hyphae and induced NF-κB activation. Mice with either blockade or genetically deleted Dectin-3 were highly susceptible to C. albicans infection. Dectin-3 constantly formed heterodimers with Dectin-2, a well-characterized CLR, for recognizing C. albicans hyphae. Compared to their respective homodimers, Dectin-3 and Dectin-2 heterodimers bound α-mannans more effectively, leading to potent inflammatory responses against fungal infections. Together, our study demonstrates that Dectin-3 forms a heterodimeric PRR with Dectin-2 for sensing fungal infection and suggests that different CLRs may form different hetero- and homodimers, which provide different sensitivity and diversity for host cells to detect various microbial infections.

Objective To clarify the mechanisms of mechanical ventilation-induced diaphragm dysfunction by exploring the effects of control mechanical ventilation(CMV) on the morphology of rat diaphragm.Methods Biopsy specimens were taken from the diaphragm after 18 h and 24 h CMV for electron microscopy and HE staining to assess muscle fiber injury;myosin heavy chain(MHC) expression and cross section area(CSA) were determined by immunohistochemistry.Results Compared with that in the control group,the density of muscle injury was significantly higher in 24 h CMV group(P0.01),and muscle fibers had irregular shape,atrophy and loose structure.Only slight changes occurred in 18 h CMV group.The positive rate of MHCfast significantly increased,but that of MHCslow tended to decrease.CSA and protein expressions of different MHC isoforms in the diaphragm were decreased(both P0.05).In 18 h CMV group,the positive rate,muscle fiber CSA as well as protein expression of MHC MHCfast or MHCslow did not alter.The density of muscle injury was positively correlated with the positive rate of MHCfast in 24 h CMV group(P0.05).Conclusion Muscle injury,muscle atrophy,and the shift of muscle fibers from slow to fast in the diaphragm occur after short-term CMV.The muscle injury may stimulate adaptive alteration of the diaphragm for improvement of muscle function.

本课题主要对2009-2010年度的针对流感病毒的四个病毒颗粒裂解疫苗(福禄立适.、华兰生物生产的流感病毒裂解疫苗、安尔来福、凡尔灵)和一个只含病毒表面糖蛋白的亚单位疫苗(爱阁力保.)进行了与免疫保护相关的有效分子的测定,同时对疫苗制备过程中可能对机体产生具有副作用的分子也加以分析.结果 显示各疫苗均能检测到神经氨酸酶活性和血凝素蛋白的红细胞凝集活性,其中安尔来福的神经氨酸酶活性高出其他产品10倍,福禄立适.则显示出最佳的红细胞凝集活性;基质蛋白仅在华兰生物生产的流感病毒裂解疫苗及安尔来福这两个裂解病毒颗粒疫苗中明显被检测到.另外,各疫苗中鸡卵清蛋白含量都很低,但安尔来福的含量明显高于其他疫苗,本研究结果可对疫苗选择提供参考.

Objective To investigate the histological alterations and the diaphragmatic expression of nitric oxide synthases (NOS) in the patients with mild-moderate chronic obstructive pulmonary disease (COPD), and to clarify the possible mechanisms of diaphragm contractile dysfunction in COPD. Methods 19 male patients undergoing thoracotomy for localized lung or esophagus neoplasm were selected for the study, 12 of them with stable mild-moderate COPD and 7 with normal pulmonary function. Diaphragm ultrastructure injuries were observed by electron microscopy. MHC isoform,cross section area (CSA) of diaphragm fibers, and the location of NOS expression in diaphragm were measured by immunohistochemistry, Western blotting was used to analyze the expression of NOS and Nitrotyrosine (NT) proteins. BMI and albumin were measured to evaluate the nutritional status, and the pulmonary function was measured before surgery. Results ?Signs of sarcomeres disruption and Z-band streaming were observed in the diaphragm of the patients with mild and moderate COPD. In addition, mitochondria of MHC isoforms were no difference between the two groups ( P＞0. 05). The CSA of the different MHC isoforms of diaphragm did not differ between the two groups, while taken the moderate COPD patients as a single group, the CSA of muscles expressed MHCslow were decreased compared with the control group ( P ＜0. 05). Compared with the control group, MHC protein expression in COPD group was significantly group, nNOS and iNOS were significantly higher in COPD group ( P＜0. 05), but eNOS expression did not differ between the two groups ( P ＞0. 05). NT expression was significantly higher compared with the control group ( P ＜0. 05). The level of nNOS isoforms and NT were inversely correlated with FEV1 %predicted. Conclusions The ultrastructure injuries and muscle atrophy occurred in the diaphragm of COPD. Endogenous NOS of diaphragm was increased significantly, which induced the increase of nitrate tyrosine. These alterations may contribute to the diaphragm dysfunction in the patients with COPD.