Distribution and significance of the oesophageal and gastric cardiac mucosae: a study of 131 operation specimens

AIMS: To clarify the distribution and significance of the oesophageal and gastric cardiac mucosae at the oesophago-gastric junction (EGJ). METHODS AND RESULTS: Oesophagectomy specimens from 131 consecutive patients with middle and upper thoracic oesophageal cancer were examined. The surgically resected specimens including the EGJ were cut into 5 mm thick serial sections and examined histopathologically for the length of the oesophageal and gastric cardiac mucosae and the incidence of columnar epithelial islands (CEIs). We also determined the presence of short-segment Barrett's oesophagus (SSBE) and goblet cell metaplasia in SSBE. Oesophageal cardiac mucosa was found in 125 cases (95%) and gastric cardiac mucosa was found in all cases. The mean length of the oesophageal and gastric cardiac mucosa was 4 mm (range 1-26 mm) and 13 mm (range 2-64 mm), respectively. CEIs were found in 75 cases (57%). SSBE was found in 70 cases (53%), among which goblet cell metaplasia was found in 28 cases (21%). No long-segment Barrett's oesophagus was found. The mean length of oesophageal cardiac mucosa (6 mm) and gastric cardiac mucosa (17 mm) in SSBE was significantly greater than that (3 mm and 8 mm, respectively) in non-SSBE cases (P < 0.0001 and P < 0.0001). The incidence (69%) of CEIs in SSBE was significantly higher than that (44%) in non-SSBE cases (P = 0.005). CONCLUSIONS: Oesophageal and gastric cardiac mucosae were found frequently. Oesophageal cardiac glands and CEIs might play an important role in the development of SSBE.     

Hypermethylation of CpG island loci and hypomethylation of LINE-1 and Alu repeats in prostate adenocarcinoma and their relationship to clinicopathological features

Promoter CpG island hypermethylation is an important carcinogenic event in prostate adenocarcinoma. Regardless of tissue type, human cancers have in common both focal CpG island hypermethylation and global genomic hypomethylation. The present study evaluated CpG island loci hypermethylation and LINE-1 and Alu repeat hypomethylation in prostate adenocarcinoma, analysed the relationship between them, and correlated these findings with clinicopathological features. We examined 179 cases of prostate adenocarcinoma and 30 cases of benign prostate hypertrophy for the methylation status of 22 CpG island loci and the methylation levels of LINE-1 and Alu repeats using methylation-specific polymerase chain reaction and combined bisulphite restriction analysis, respectively. The following 16 CpG island loci were found to display cancer-related hypermethylation: RASSF1A, GSTP1, RARB, TNFRSF10C, APC, BCL2, MDR1, ASC, TIG1, RBP1, COX2, THBS1, TNFRSF10D, CD44, p16, and RUNX3. Except for the last four CpG island loci, hypermethylation of each of the remaining 12 CpG island loci displayed a close association with one or more of the prognostic parameters (ie preoperative serum prostate specific antigen level, Gleason score sum, and clinical stage). Prostate adenocarcinoma with hypermethylation of each of ASC, COX2, RARB, TNFRSF10C, MDR1, TIG1, RBP1, NEUROG1, RASSF1A, and GSTP1 showed a significantly lower methylation level of Alu or LINE-1 than prostate adenocarcinoma without hypermethylation. In addition, hypomethylation of Alu or LINE-1 was closely associated with one or more of the above prognostic parameters. These data suggest that in tumour progression a close relationship exists between CpG island hypermethylation and the hypomethylation of repetitive elements, and that CpG island hypermethylation and DNA hypomethylation contribute to cancer progression.     

沙门菌致病岛1蛋白促进神经母细胞瘤Warburg效应的作用及分子机制

目的探讨沙门菌致病岛1(SPI1)蛋白在神经母细胞瘤中调控有氧糖酵解(即Warburg效应)的作用及分子机制。方法运用公用数据库,解析神经母细胞瘤中调控Warburg效应的候选关键转录因子SPI1;经嘌呤霉素筛选,获得稳定过表达/敲低SPI1的SK-N-BE(2)细胞株,分为空载体对照(Mock)组、SPI1过表达(SPI1)组、随机干扰(sh-Scb)组、SPI1干扰(sh-SPI1)组;实时荧光定量聚合酶链反应(PCR)、染色质免疫共沉淀、双荧光素酶报告基因及蛋白质印迹法(Western blot)检测靶基因启动子活性和表达水平;分光光度计法和糖酵解压力试验检测有氧糖酵解水平;软琼脂克隆形成和基质胶侵袭检测瘤细胞的增殖和侵袭活性。组间比较采用t检验。结果SPI1组己糖激酶2(HK2)表达高于Mock组(3.63±0.53比1.00±0.05,t=17.462,P<0.05),磷酸甘油酸激酶1(PGK1)表达高于Mock组(3.28±0.39比1.00±0.03,t=17.651,P<0.05),细胞外酸化率(ECAR)高于sh-Scb组[(32.17±3.15)mpH/min比(9.82±1.49)mpH/min,t=10.352,P<0.05],克隆形成数高于Mock组[(163.30±7.59)个比(48.02±4.85)个,t=17.840,P<0.05],侵袭细胞数高于Mock组[(184.70±10.58)个比(88.33±5.56)个,t=8.214,P<0.05]。sh-SPI1组HK2表达低于sh-Scb组(0.33±0.02比1.00±0.04,t=14.370,P<0.05),PGK1表达低于sh-Scb组(0.34±0.02比1.00±0.03,t=19.192,P<0.05),ECAR值低于sh-Scb组(3.07±0.48比10.05±1.37,t=7.580,P<0.05),克隆形成数低于sh-Scb组[(21.33±3.96)个比(44.67±5.69)个,t=5.916,P<0.05],侵袭细胞数低于sh-Scb组[(40.67±3.48)个比(88.00±6.89)个,t=4.994,P<0.05],差异均有统计学意义。结论转录因子SPI1通过增强HK2和PGK1表达,促进神经母细胞瘤的Warburg效应和肿瘤进展。     

1995-2005年海岛地区性病疫情分析

目的 分析浙江省玉环县1995-2005年11年性传播疾病(STD)的流行趋势及特点,为制定有效的防治策略提供依据.方法 对玉环县11年8种法定报告STD监测资料进行统计分析.结果 11年8种监测STD中,除性病性淋病肉芽肿外,其余7种均有报告,累计报告STD 8479例,年平均发病率197.47/10万,年平均增长率为12.68%.结论 玉环县STD发病呈增长趋势,其中梅毒和非淋球菌性尿道炎(NGU)的增长较为显著,加强STD防治知识的普及,提倡安全性行为,是控制STD蔓延的有效措施.     

指动脉顺行V-Y岛状皮瓣修复指腹缺损

目的评价指动脉顺行"V-Y"岛状皮瓣修复手指指腹缺损的临床疗效。方法2015年3月-2018年6月,应用指动脉顺行"V-Y"岛状皮瓣修复手指指腹缺损20例,均有末节指骨外露,缺损面积最小1.0 cm×1.0 cm,最大1.5 cm×1.5 cm,供区均直接闭合。结果20例皮瓣全部成活,皮瓣及手指外形满意。随访3~6个月,平均4.5个月,两点辨别觉3~5 mm。指间关节屈曲活动18例均无明显受限,2例因手指屈曲位缝合皮瓣,出现手指远指间关节屈伸轻度受限。按TAM法评定标准评定:优良率90%。结论顺行指动脉"V-Y"岛状皮瓣手术操作不仅简单,同时内含血管神经束,血供稳定,感觉恢复良好,能够恢复良好的手指外形和功能,是修复指腹缺损的比较理想的手术方式之一。     

Fifty million years of beetle evolution along the Antarctic Polar Front

Global cooling and glacial–interglacial cycles since Antarctica’s isolation have been responsible for the diversification of the region’s marine fauna. By contrast, these same Earth system processes are thought to have played little role terrestrially, other than driving widespread extinctions. Here, we show that on islands along the Antarctic Polar Front, paleoclimatic processes have been key to diversification of one of the world’s most geographically isolated and unique groups of herbivorous beetles—Ectemnorhinini weevils. Combining phylogenomic, phylogenetic, and phylogeographic approaches, we demonstrate that these weevils colonized the sub-Antarctic islands from Africa at least 50 Ma ago and repeatedly dispersed among them. As the climate cooled from the mid-Miocene, diversification of the beetles accelerated, resulting in two species-rich clades. One of these clades specialized to feed on cryptogams, typical of the polar habitats that came to prevail under Miocene conditions yet remarkable as a food source for any beetle. This clade’s most unusual representative is a marine weevil currently undergoing further speciation. The other clade retained the more common weevil habit of feeding on angiosperms, which likely survived glaciation in isolated refugia. Diversification of Ectemnorhinini weevils occurred in synchrony with many other Antarctic radiations, including penguins and notothenioid fishes, and coincided with major environmental changes. Our results thus indicate that geo-climatically driven diversification has progressed similarly for Antarctic marine and terrestrial organisms since the Miocene, potentially constituting a general biodiversity paradigm that should be sought broadly for the region’s taxa.

Antarctica’s isolation, cooling, and glacial–interglacial cycles over the Cenozoic have resulted in the remarkable diversification of a unique marine fauna (1, 2). The investigation of marine radiations in Antarctica has reshaped modern understanding of biodiversity processes, for example, by revealing a surprising inverse latitudinal gradient in diversification rates for fish and brittle stars (3–5). In contrast, Antarctica’s paleoclimatic legacy for terrestrial communities has long been considered one of widespread extinction due to glaciation. Evidence of terrestrial species surviving in Antarctic glacial refugia (6) and discoveries of substantial endemic diversity and biogeographic structuring in some groups (7, 8) is changing this narrative, indicating extended evolutionary histories on land. Yet, such evolutionary histories remain obscured by a lack of large-scale molecular phylogenetic work, with most Antarctic terrestrial research focused on small subsets of species or populations (9, 10). The few studies that have taken a multilocus phylogenetic approach have uncovered hidden terrestrial diversity and signals of long-term allopatric divergence (e.g., refs. 11 and 12), hinting that Cenozoic climatic processes may have driven terrestrial diversification in ways similar to that for marine life.The hypothesis that diversification has proceeded similarly in Antarctic marine and terrestrial groups has not been tested. While the extinction of a diverse continental Antarctic biota is well established (13), mounting evidence of significant and biogeographically structured Antarctic terrestrial diversity (8, 14, 15) with a long evolutionary history (6, 16) suggests the possibility of broadly similar diversification processes across marine and terrestrial Antarctic systems. If valid for some taxa, further tests should then be sought across a wider variety of organisms. Here, we therefore evaluate the terrestrial applicability of the paradigm emerging for Antarctic marine biodiversity—that a major cooling phase from the mid-Miocene climatic transition (14 Ma) onwards, and subsequent habitat restructuring, have led to significant and ongoing diversification for many taxa, including those with much older origins in the region (2, 4, 17). We do so by using one of the most well-known and speciose groups from the sub-Antarctic, the herbivorous Ectemnorhinini weevils (Coleoptera: Curculionidae) (18–20).Preliminary molecular studies indicate that the Ectemnorhinini, along with numerous other terrestrial taxa, have long histories in the sub-Antarctic, extending to the Miocene or earlier [e.g., beetles (21), midges (22), and springtails (11)]. This enables a comparison of their evolution throughout the same periods of environmental change that drove the diversification of Antarctic marine taxa. Moreover, the sub-Antarctic islands overlap spatially with the Southern Ocean, with climates that reflect oceanic conditions both past and present (23, 24). While in some respects quite different to the continental Antarctic, the islands are in other ways quite similar, providing a window into diversification processes that might be sought for continental groups, especially given their age and biogeographic structuring. Both regions share many higher taxa (e.g., ref. 25), a dynamic geo-climatic history (6, 26), a profound degree of isolation, and indications that climatic events likely structured their biota (6, 8, 27). The terrestrial habitat on the continent and its surrounding islands is fragmented by large expanses of ice or ocean, respectively, and has been further isolated by the Antarctic Circumpolar Current for at least 34 Ma (28, 29). Cyclic growth and contraction of ice sheets throughout the Plio–Pleistocene, though typically associated with the continent, has also had extensive impacts on the sub-Antarctic islands (26). The more intensively surveyed sub-Antarctic faunas thus provide an opportunity to investigate terrestrial diversification processes for the wider Antarctic while recognizing that for many groups on the continent, the main legacy of change has been extinction.To test the hypothesis that a major phase of cooling from the mid-Miocene onwards and subsequent habitat restructuring has led to the diversification of Antarctic terrestrial taxa, we integrate three tiers of molecular data to reveal a comprehensive evolutionary history for the Ectemnorhinini weevils. This additionally allows us to resolve the geographic, taxonomic, and temporal origins of the Ectemnorhinini and the role of dispersal and colonization in the development of the region’s biogeography. We first resolve the controversial origins of these weevils (19, 30) with a phylogenomic approach using anchored hybrid enrichment (AHE) for up to 515 genes across 12 representative species of Ectemnorhinini and a worldwide sample of 87 species of putative relatives and known outgroups, mostly from the beetle subfamily Entiminae (18, 30, 31). We then build on these outcomes by exploring the timing and patterns of taxonomic diversification, including divergence times and proposed dispersal events, using a multilocus phylogenetic dataset (three mitochondrial and two nuclear genes) for an extensive sample of Ectemnorhinini from each archipelago on which they are known to occur. Finally, we reveal contemporary limits to gene flow and examine the population structure of the littoral-dwelling ectemnorhinine weevil Palirhoeus eatoni using phylogeographic methods applied to a library of 5,859 genome-wide single-nucleotide polymorphisms (SNPs). This unusually widespread species is found on all four archipelagos of the Kerguelen Province known to host Ectemnorhinini: Crozet, Kerguelen, Prince Edward Islands (PEI), and Heard Island and McDonald Islands (HIMI).     

5种食源性致病微生物毒力基因重组质粒的构建、克隆表达与表达产物的研究

目的构建大肠杆菌O157∶H7、副溶血性弧菌、金黄色葡萄球菌、沙门氏菌和志贺氏菌5种主要食源性致病微生物毒力基因重组质粒载体并鉴定其表达产物,为探讨高通量磁性荧光纳米颗粒标记相应的抗体技术快速检测带毒力基因的致病微生物的可行性奠定基础。方法分别从5种食源性致病微生物毒力岛中选择特异性的毒力基因进行引物设计,分别提取5种目的细菌DNA片段,经PCR扩增、电泳回收目的基因片段,将目的基因片段与原核表达载体pET-23a、pET-28a、pET-32a构建各自的重组质粒,将重组质粒转化入大肠杆菌DH5α内并提取质粒载体,构建后的重组质粒进行酶切和测序鉴定,再转化入表达宿主E·coliBL21（DE3）。在0.1 mmol/L的IPTG诱导下,目的质粒载体在E·coliBL21（DE3）株中表达,SDS-PAGE电泳检测表达蛋白。结果实验成功构建大肠杆菌O157∶H7、副溶血性弧菌、金黄色葡萄球菌、沙门氏菌和志贺氏菌等细菌的毒力基因重组质粒载体pET-23a-eaeA、pET-28a-tdh、pET-28a-enterotoxin B、pET-32a-invA和pET-28a-ipaH,并克隆出969 bp的eaeA基因片段、567 bp的tdh基因片段、798 bp的enterotoxin B基因片段、1 041 bp的invA基因片段和771 bp的ipaH基因片段。重组质粒载体经酶切和测序与目标基因序列一致。在E·coliBL21（DE3）株中有质粒表达蛋白37.5 kDa（eaeA）、26 kDa（tdh）、34.5 kDa（enterotoxin B）、41 kDa（invA）、32 kDa（ipaH）。结论本研究成功构建了5种食源性致病微生物毒力基因重组质粒并在原核细胞中高效表达,为下一步获得相应的毒力基因抗体及快速诊断试剂的研制应用奠定了基础。