我国农作物重大迁飞性害虫发生为害及监测预报技术

文章概述了我国一类农作物病虫害中7种(类)迁飞性害虫发生为害概况,即是我国三大粮食作物上为害最为严重的害虫,发生区域涵盖我国粮食作物种植区,且存在国内外虫源交流;概述了成虫种群监测、雌蛾卵巢发育分级、翅型调查、异地预报等迁飞性害虫监测预报技术;提出了需要对迁飞性害虫成灾机理、智能化监测工具和迁飞关键路径进行研究,努力建成全国空天地一体化监测网络,实现迁飞性害虫的精准预报的建议.     

Electron cryotomography of ESCRT assemblies and dividing Sulfolobus cells suggests that spiraling filaments are involved in membrane scission

The endosomal-sorting complex required for transport (ESCRT) is evolutionarily conserved from Archaea to eukaryotes. The complex drives membrane scission events in a range of processes, including cytokinesis in Metazoa and some Archaea. CdvA is the protein in Archaea that recruits ESCRT-III to the membrane. Using electron cryotomography (ECT), we find that CdvA polymerizes into helical filaments wrapped around liposomes. ESCRT-III proteins are responsible for the cinching of membranes and have been shown to assemble into helical tubes in vitro, but here we show that they also can form nested tubes and nested cones, which reveal surprisingly numerous and versatile contacts. To observe the ESCRT–CdvA complex in a physiological context, we used ECT to image the archaeon Sulfolobus acidocaldarius and observed a distinct protein belt at the leading edge of constriction furrows in dividing cells. The known dimensions of ESCRT-III proteins constrain their possible orientations within each of these structures and point to the involvement of spiraling filaments in membrane scission.     

分子伴侣及信号肽对毕赤酵母中分泌蛋白表达水平的影响

目的：探索定位于细胞质、内质网膜及内质网腔中的分子伴侣及其组合对于带有不同信号肽的胞外β-1,3-葡聚糖酶（EXGl）在巴斯德毕赤酵母GS200中表达水平的影响。方法：通过融合PCR技术分别构建带有酵母a交配因子引导肽序列（仅MF）、酵母仅交配因子信号肽序列（ccPre）和重链结合蛋白（Bip）信号肽序列的报告蛋白EXGl的表达质粒pPIC9-EXG1,同时构建分子伴侣基因及其组合的表达质粒pBLArg-IV,然后将2种重组质粒共转化至毕赤酵母宿主菌GS200,转化子经筛选获得共表达菌株,通过测定EXG1酶活来评价分子伴侣与信号肽对其表达水平的影响。结果：细胞质及内质网膜上的分子伴侣Sec61a、Sec61B及胞质中的分子伴侣Ydjl、Ssal、Hsp104及其组合对各种信号肽引导的报告蛋白EXG1的表达水平没有显著影响。然而,内质网腔中的分子伴侣Bip、EroI、PDI与HacI组合能显著提高报告蛋白EXG1的表达水平,其中,以aMF或ctPre作为信号肽引导的报告蛋白EXG1的表达水平分别提高了2．6倍和3．8倍,以Bip信号肽引导的报告蛋白EXGl的表达水平提高了20％～45％,而对于以EXG1自身信号肽引导的报告蛋白EXG1的表达水平没有显著影响。结论：在酵母表达体系中,内质网腔中的分子伴侣是报告蛋白EXG1表达水平的重要影响因素．但分子伴侣对于信号肽的选择性还须进一步证明。     

A Relationship Between Replication Protein A and Occurrence and Prognosis of Esophageal Carcinoma

Esophageal carcinoma (EC) is an aggressive and the third most common cancer of the digestive tract with poor prognosis. Replication protein A (RPA) is critically required for DNA replication and its elevated expression has been observed in many malignant tumors. In this study, we investigated the expression of RPA1 and RPA2, subunits of RPA, and assessed their prognostic value in EC patients. We analyzed immunohistochemically the expression of RPA1 and RPA2 proteins in 48 EC resection specimens in relation with clinicopathological parameters and survival. We observed a significant elevated (P < 0.001) RPA1 and RPA2 expressions (labeling index) in the tumor than adjacent non-tumor tissues. In addition, both RPA1 and RPA2 labeling index in lymph node metastasis patients was significantly higher (both P = 0.000) than patients without lymph node metastasis. However, RPA1 and RPA2 labeling index in early stage was significantly lower (P = 0.000 and P = 0.002, respectively) than that of late stage EC patients. Importantly, patient’s survival at early stage was significantly higher (P = 0.016) than late stage EC and lymph node metastasis and RPA1 expression was associated with adverse patient’s outcome in multivariate analysis (P < 0.05 and P < 0.00, respectively). In conclusion, RPA1 could be a useful prognostic indicator in patients with esophageal carcinoma and might be a future attractive therapeutic target for regulation by tumor suppressors.     

The influence of cathelicidin LL37 in human anti-neutrophils cytoplasmic antibody (ANCA)-associated vasculitis

Introduction

Antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) is characterised by the autoinflammation and necrosis of blood vessel walls. The renal involvement is commonly characterised by a pauci-immune crescentic glomerulonephritis (PiCGN) with a very rapid decline in renal function. Cathelicidin LL37, an endogenous antimicrobial peptide, has recently been implicated in the pathogenesis of autoimmune diseases. To assess whether serum LL37 reflects renal crescentic formation, we measured the serum levels of LL37 in AAV patients with and without crescentic glomerulonephritis (crescentic GN) as compared to healthy controls (HCs). We also analysed the correlation of the serum levels of LL37 and interferon-α (IFN-α) with the clinical characteristics of the patients.

Methods

The study population consisted of 85 AAV patients and 51 HCs. In 40 ANCA-positive patients, a parallel analysis was performed, including the assessment of LL37 and IFN-α levels in the serum and renal biopsies. Of those studied, 15 AAV patients had biopsy-proven crescentic GN, and 25 AAV patients lacked crescent formation. The serum levels of cathelicidin LL37 and IFN-α were both measured by ELISA, and the clinical and serological parameters were assessed according to routine procedures. Immunofluorescence staining was performed on frozen sections of kidney needle biopsies from AAV patients with crescentic GN.

Results

The serum levels of LL37 and IFN-α were significantly increased in AAV patients with crescentic GN compared to AAV patients without crescentic formation and HCs, and patients with high LL37 and IFN-α levels were more likely to be in the crescentic GN group. The LL37 levels were positively correlated with the IFN-α levels, and both LL37 and IFN-α levels showed a positive correlation with serum creatinine and no correlation with complement C3. The renal tissue of crescentic GN patients showed expression of LL37 and IFN-α at the Bowman’s capsule and extracellular sites, suggesting the active release of LL37 and IFN-α.

Conclusions

Significantly higher levels of LL-37 and IFN-α were observed in AAV patients, particularly those with crescentic formation, and LL37 and IFN-α were expressed in the renal tissue of patients with crescentic GN. These data suggest that serum levels of LL37 and IFN-α may reflect both local renal inflammation and systemic inflammation.     

Efficient secretion of lipase r27RCL in Pichia pastoris by enhancing the disulfide bond formation pathway in the endoplasmic reticulum

The lipase r27RCL from Rhizopus chinensis CCTCC M201021 was heterologously expressed in Pichia pastoris GS115 by simultaneous co-expression with two secretion factors ERO1p and PDI involved in the endoplasmic reticulum (ER). Compared to the expression of the lipase alone (12,500 U/ml), co-expression with these two proteins resulted in the production of larger total quantities of enzymes. The largest increase was seen when the combined ERO1p/PDI system was co-expressed, resulting in approximately 30 % higher enzyme yields (16,200 U/ml) than in the absence of co-expressed secretion factors. The extracellular protein concentration of the recombinant strain Co XY RCL-5 reached 9.39 g/l in the 7-l fermentor. Simultaneously, the fermentation time was also shortened by about 8 h compared to that of the control. The substrate-specific consumption rate (Qs) and the product-specific production rate (Qp) were both investigated in this research. In conclusion, the space–time yield was improved by co-expression with ERO1p and PDI. This is a potential strategy for high level expression of other heterologous proteins in P. pastoris.     

PICK1 and ICA69 Control Insulin Granule Trafficking and Their Deficiencies Lead to Impaired Glucose Tolerance

Diabetes is a metabolic disorder characterized by hyperglycemia. Insulin, which is secreted by pancreatic beta cells, is recognized as the critical regulator of blood glucose, but the molecular machinery responsible for insulin trafficking remains poorly defined. In particular, the roles of cytosolic factors that govern the formation and maturation of insulin granules are unclear. Here we report that PICK1 and ICA69, two cytosolic lipid-binding proteins, formed heteromeric BAR-domain complexes that associated with insulin granules at different stages of their maturation. PICK1-ICA69 heteromeric complexes associated with immature secretory granules near the trans-Golgi network (TGN). A brief treatment of Brefeldin A, which blocks vesicle budding from the Golgi, increased the amount of PICK1 and ICA69 at TGN. On the other hand, mature secretory granules were associated with PICK1 only, not ICA69. PICK1 deficiency in mice caused the complete loss of ICA69 and led to increased food and water intake but lower body weight. Glucose tolerance tests demonstrated that these mutant mice had high blood glucose, a consequence of insufficient insulin. Importantly, while the total insulin level was reduced in PICK1-deficient beta cells, proinsulin was increased. Lastly, ICA69 knockout mice also displayed similar phenotype as the mice deficient in PICK1. Together, our results indicate that PICK1 and ICA69 are key regulators of the formation and maturation of insulin granules.

Author Summary

Insulin is a key regulator of blood glucose and insufficient insulin leads to diabetes. Insulin is synthesized as proinsulin, processed in endoplasmic reticulum and Golgi, and eventually packaged into insulin granules, a type of dense core vesicles. Despite its importance, the molecular mechanisms governing the biogenesis and maturation of insulin granules are not fully understood. In this study, we identified two cytosolic proteins, PICK1 and ICA69, as important regulators of insulin granule biogenesis and maturation. Both PICK1 and ICA69 have the banana-shaped BAR domain that can bend the lipid membrane and help the formation of dense core vesicles. We show that without PICK1 or ICA69, insulin granules cannot be properly formed and, as a result, proinsulin cannot be effectively processed into mature insulin. Mice lacking functional PICK1 or ICA69 genes have reduced insulin but increased proinsulin. Consequently, these mice have high levels of glucose, a prominent feature found in diabetes patients. These results add to previous findings that PICK1 is important for the generation of proacrosomal granules found in cells of the testis, and thereby support a wider role for PICK1 and ICA69 in regulating dense core vesicle biogenesis and maturation.     

Evaluation of Five Candidate Genes from GWAS for Association with Oligozoospermia in a Han Chinese Population

Background

Oligozoospermia is one of the severe forms of idiopathic male infertility. However, its pathology is largely unknown, and few genetic factors have been defined. Our previous genome-wide association study (GWAS) has identified four risk loci for non-obstructive azoospermia (NOA).

Objective

To investigate the potentially functional genetic variants (including not only common variants, but also less-common and rare variants) of these loci on spermatogenic impairment, especially oligozoospermia.

Design, Setting, and Participants

A total of 784 individuals with oligozoospermia and 592 healthy controls were recruited to this study from March 2004 and January 2011.

Measurements

We conducted a two-stage study to explore the association between oligozoospermia and new makers near NOA risk loci. In the first stage, we used next generation sequencing (NGS) in 96 oligozoospermia cases and 96 healthy controls to screen oligozoospermia-susceptible genetic variants. Next, we validated these variants in a large cohort containing 688 cases and 496 controls by SNPscan for high-throughput Single Nucleotide Polymorphism (SNP) genotyping.

Results and Limitations

Totally, we observed seven oligozoospermia associated variants (rs3791185 and rs2232015 in PRMT6, rs146039840 and rs11046992 in Sox5, rs1129332 in PEX10, rs3197744 in SIRPA, rs1048055 in SIRPG) in the first stage. In the validation stage, rs3197744 in SIRPA and rs11046992 in Sox5 were associated with increased risk of oligozoospermia with an odds ratio (OR) of 4.62 (P  =  0.005, 95%CI 1.58-13.4) and 1.82 (P  =  0.005, 95%CI 1.01-1.64), respectively. Further investigation in larger populations and functional characterizations are needed to validate our findings.

Conclusions

Our study provides evidence of independent oligozoospermia risk alleles driven by variants in the potentially functional regions of genes discovered by GWAS. Our findings suggest that integrating sequence data with large-scale genotyping will serve as an effective strategy for discovering risk alleles in the future.     

A Novel PRPF31 Mutation in a Large Chinese Family with Autosomal Dominant Retinitis Pigmentosa and Macular Degeneration

Purpose

This study was intended to identify the disease causing genes in a large Chinese family with autosomal dominant retinitis pigmentosa and macular degeneration.

Methods

A genome scan analysis was conducted in this family for disease gene preliminary mapping. Snapshot analysis of selected SNPs for two-point LOD score analysis for candidate gene filter. Candidate gene PRPF31 whole exons'' sequencing was executed to identify mutations.

Results

A novel nonsense mutation caused by an insertion was found in PRPF31 gene. All the 19 RP patients in 1085 family are carrying this heterozygous nonsense mutation. The nonsense mutation is in PRPF31 gene exon9 at chr19:54629961-54629961, inserting nucleotide “A” that generates the coding protein frame shift from p.307 and early termination at p.322 in the snoRNA binding domain (NOP domain).

Conclusion

This report is the first to associate PRPF31 gene''s nonsense mutation and adRP and JMD. Our findings revealed that PRPF31 can lead to different clinical phenotypes in the same family, resulting either in adRP or syndrome of adRP and JMD. We believe our identification of the novel “A” insertion mutation in exon9 at chr19:54629961-54629961 in PRPF31 can provide further genetic evidence for clinical test for adRP and JMD.