Pulmonary neuroendocrine cells sense succinate to stimulate myoepithelial cell contraction

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Differentiation of immortalized epithelial cells derived from cystic fibrosis airway submucosal glands

Summary Cystic fibrosis (CF) involves abnormalities in mucus production and secretion of the airway. Studies of the regulation of airway mucin production and secretion has been difficult due to the lack of in vitro models of the airway epithelial cells which express functional differentiation. Because the majority of the mucin in the airway is apparently produced by the submucosal glands, we have focused our attention on the development of cell culture models of human airway submucosal glands. This report describes the propagation of CF airway submucosal gland epithelial cells which continue to express mucin production. The CF bronchus was obtained from a 31-yr-old patient who received a double lung transplant. The glands were dissected out and primary cultures prepared by the explant/outgrowth procedure. The cells were immortalized by infection with Adl2-SV40 hybrid virus. The cultures are maintained in serum-free keratinocyte basal medium supplemented with insulin (5μg/ml), hydrocortisone (0.5μg/ml), epidermal growth factor (10 ng/ml), bovine pituitary extract (25μg/ml), and antibiotics. Cultures were passaged using 0.125% trypsin in Ca⁺² and Mg⁺²-free Hanks’, balanced salt solution. Polymerase chain reaction (PCR) analysis demonstrated that the cells were homozygous for the ΔF508 mutation. Morphologic observations showed that the cells were epithelial and were interconnected by sparsely distributed desmosomes. Their cytoplasm contained secretory-type structures including abundant Golgi, rough endoplasmic reticulum, and secretory vesicles. Immunofluorescent studies determined that all cells were positive for cytokeratins, mucin glycoconjugates, and cystic fibrosis transmembrane conductance regulator. The cultures secreted substantial amounts of mucin glycoproteins and expressed the MUC-2 mucin gene. Patch clamp experiments revealed that the cells expressed defective Cl⁻ channels which were not activated by Forskolin.     

生防菌株HG18基因组信息分析与抗菌功能基因挖掘

贝莱斯芽胞杆菌（Bacillus velezensis）HG18是1株低温生防菌株,能够分泌抗菌物质。为挖掘和利用其抗菌功能基因,服务农业生产,采用二、三代相结合测序技术,对其进行全基因组测序,获得菌株完整基因组序列。基因组全长4 461 844 bp,包含一个染色体和一个质粒,GC含量44.06%,编码4 643个基因,编码基因总长度3 893 994 bp,占基因组87.27%。发现6个几丁质降解相关基因,2个葡聚糖酶基因和1个壳聚糖酶基因,2个脂肽类抗菌物质芬芥素与表面活性素合成基因簇,2个细菌素subtilin和bacillolysin合成基因。研究为提高抗菌物质产量的菌株定向遗传改造以及植物抗病育种提供基因资源。     

Adipose tissue-derived stem cells upon decellularized ovine small intestine submucosa for tissue regeneration: An optimization and comparison method

The extracellular matrix of different mammalian tissues is commonly used as scaffolds in the field of tissue engineering. One of these tissues, which has frequently been studied due to its structural and biological features, is the small intestine submucosal membrane. These research are mainly done on the porcine small intestine. However, a report has recently been published about a scaffold produced from the submucosal layer of the ovine small intestine. In the present study, ovine small intestine submucosal (OSIS) was decellularized in a modified manner and its histological, morphological, and biomechanical properties were studied. Decellularization was performed in two phases: physical and chemical. In this method, a chloroform-methanol mixture, enzymatic digestion, and a constant dose of sodium dodecyl sulfate (SDS) was used in the least agitation time and its histological property and biocompatibility were evaluated in the presence of adipose tissue-derived stem cells (ADSCs); furthermore, ADSCs were isolated with a simple method (modified physical washing non-enzymatic isolation). The results were showed that the use of OSIS could be effective and operative. Mechanical properties, histological structure and shape, and glycosaminoglycan content were preserved. In the SDS-treated group, more than 90% of the native cells of tissue were deleted, and also in this group, no toxicity was observed and cell proliferation was supported, compared to the untreated group. Therefore, our results indicate that ADSCs seeded on OSIS scaffold could be used as a new approach in regenerative medicine as hybrid or hydrogel application.     

The response of meiofauna and microphytobenthos to engineering effects of fiddler crabs on a subtropical intertidal sandflat

Fiddler crabs are key bioturbators on tidal flats. During their intense bioturbation process, they manipulate large amounts of sediment, altering the physical state of existing materials. We investigated whether different types of sediment bioturbation produced by fiddler crabs modulate meiofaunal assemblages and microphytobenthic content. We hypothesized that sedimentary structures produced by burrowing (the burrow itself and the excavation pellets) and feeding (feeding pellets) generate different microenvironments compared with areas without apparent signs of fiddler crab disturbance, affecting both meiofauna and microphytobenthos, independent of the sampling period. Our results indicate that the engineering effects of burrow construction and maintenance and the engineering effects of fiddler crab foraging modulate meiofaunal assemblages in different ways. Overall, meiofauna from burrows and excavation pellets was more abundant and diverse than at control sites, whereas feeding pellets contained poor meiofaunal assemblages. By contrast, only foraging effects were detected on microphytobenthos; independent of the sampling period, Chl a and phaeopigment content were higher in the feeding pellets, but similar among burrows, excavation pellets and control sites. The present study demonstrates that the different engineering effects of fiddler crabs are an important source of habitat heterogeneity and a structuring agent of meiofaunal assemblages on subtropical tidal flats.     

内镜黏膜下剥离术治疗消化系统早癌及癌前病变的临床疗效

目的:探讨内镜黏膜下剥离术(ESD)对消化道早癌及癌前病变的治疗效果。方法:选择2013年8月至2014年8月在我院接受治疗的消化道肿瘤患者79例作为研究对象,根据手术方法不同将所选患者分为ESD组(49例)和对照组(30例)。ESD组患者采用内镜黏膜下剥离术治疗,对照组采用传统手术治疗。观察并比较两组患者的手术时间、治愈性切除率、整块完整切除率、术后并发症的发生率及复发、转移情况。结果:ESD组患者的手术时间少于对照组,差异具有统计学意义(P0.05);两组患者手术治愈性切除率均为100%,差异无统计学意义(P0.05);ESD组手术整块完整切除率(63.27%)低于对照组(86.67%),差异具有统计学意义(P0.05)。ESD组患者术后并发症的发生率(4.08%)显著低于对照组(13.3%),差异具有统计学意义(P0.05)。两组患者术后一年内均未出现原发病灶转移及复发。结论:ESD治疗消化道早癌及癌前病变的临床疗效较好,与传统手术相比,ESD手术并发症较少、且安全性较高,更加适宜临床推广及应用。     

内镜黏膜下剥离术和挖除术治疗上消化道肿瘤的疗效及安全性分析

目的:分析内镜黏膜下剥离术(ESD)和黏膜下挖除术(ESE)治疗上消化道肿瘤的疗效及安全性。方法:回顾性分析2017年1月至2019年4月我院消化内科接受ESD或ESE治疗的68例上消化道肿瘤住院患者的临床资料,收集患者基础疾病、手术时间、病变部位大小、整块切除率、并发症等资料,同时采用Logistic回归分析对术中穿孔进行危险因素分析。结果:64例患者完整切除瘤体(94.12%),肿瘤平均直径(16.98±8.29)mm。食管病灶病理类型以高级别上皮内瘤变为主,有15例(22.06%);胃部病灶分布以胃体、胃底和胃窦居多,分别有18例(26.47%)、16例(23.53%)和12例(17.65%),病理类型以间质瘤最多,占36.76%;11例患者发生并发症(16.18%),4例患者出现术后出血(5.88%),经过内镜下止血后好转,8例患术中穿孔(11.76%),均行内镜下尼龙绳联合钛夹行荷包缝合,其中1例患者术中穿孔合并术后迟发性出血;病灶最大直径(≥25 mm,P=0.036)和病灶部位(胃底,P=0.015)是导致ESD或ESE术中穿孔的独立危险因素。结论:ESD和ESE治疗上消化道肿瘤安全有效,但需注意病灶大小和胃底病变,因其是导致术中穿孔的独立危险因素。     

超声内镜辅助下内镜黏膜下切除术对食管癌前病变患者肿瘤标志物及应激反应指标的影响

目的:探讨超声内镜(EUS)辅助下内镜黏膜下切除术(EMR)对食管癌前病变患者肿瘤标志物及应激反应指标的影响。方法:选择山东大学齐鲁医院青岛院区消化内科于2016年3月至2018年4月期间收治的食管癌前病变患者137例,采用随机数字表法将患者分为常规组(n=68,常规胃镜下行EMR)和EUS组(n=69,EUS辅助下行EMR),比较两组患者临床指标,比较两组术前、术后血清肿瘤标志物及应激反应指标水平,比较两组术前、术后1周相关遗传学分子水平。结果:EUS组手术时间、术后流质饮食时间均短于常规组(P0.05),并发食管黏膜小穿孔例数、使用钛夹止血例数均少于常规组(P0.05)。两组患者术后肿瘤特异性生长因子(TSGF)、细胞角蛋白19血清片段21-1(CYFRA21-1)、鳞状上皮细胞癌抗原(SCC-Ag)均较术前升高,但EUS组低于常规组(P0.05)。两组患者术后肾上腺素(E)、去甲肾上腺素(NE)、肾素(R)、血管紧张素Ⅱ(AngⅡ)、醛固酮均较术前升高,但EUS组低于常规组(P0.05)。两组患者术后1周细胞周期素E(Cyclin E)、转化生长因子-α(TGF-α)均较术前降低,且EUS组低于常规组(P0.05)。结论:相比于常规胃镜,经EUS辅助下EMR治疗食管癌前病变可有效改善患者的临床指标,减轻患者应激反应,有利于降低血清肿瘤标志物及遗传学分子水平。     

两种内镜手术对消化道神经内分泌肿瘤的切除效果及安全性评价

目的:探讨内镜下两种手术方法对消化道神经内分泌肿瘤的切除效果和安全性。方法:选取64例消化道神经内分泌肿瘤患者,随机分为A组和B组,每组各32例。A组接受内镜下黏膜切除术,B组接受内镜下黏膜剥离术。分析和比较两组所切除组织的病理学检查结果、手术时间、切除肿瘤的直径和厚度、治疗费用、住院时间、肿瘤完全切除率、垂直切缘阴性率以及并发症的发生率。结果:A组的手术时间为(8.95±1.63) min,治疗费用为(2127.70±468.31)元,均显著少于B组(P0.05);两组患者切除肿瘤直径和厚度、住院时间、垂直切缘阴性率对比差异均没有统计学意义(P0.05);B组的肿瘤完全切除率为93.75%,显著高于A组(P0.05);A组并发症发生率为3.13%,显著低于B组(P0.05)。结论:两种内镜下手术方式均可有效清除消化道神经内分泌肿瘤病灶。内镜下黏膜切除术的手术时间、费用及并发症的发生率更少低,而内镜下黏膜剥离术能够更彻底地清除肿瘤组织。     

老年胆总管结石患者ERCP治疗前后炎性因子、免疫功能的变化及术后并发胰腺炎的危险因素研究

摘要 目的：观察老年胆总管结石患者经内镜逆行胰胆管造影术（ERCP）治疗后,炎性因子、免疫功能的变化,并分析术后并发胰腺炎的危险因素。方法：纳入2015年3月~2021年5月期间来我院接受治疗的老年胆总管结石患者250例,均接受ERCP治疗,观察治疗前后炎性因子、免疫功能的变化,记录两组一次性取石成功率、术中出血量、手术时间、住院时间及术后并发症发生率。采用多因素Logistic回归分析术后并发胰腺炎的危险因素。结果：本次研究纳入的患者一次性取石成功率为96.80%（242/250）,术中出血量（16.57±1.34）ml,手术时间（46.38±5.19）min,住院时间（8.82±1.35）d。术后并发症发生率12.80%（32/250）。治疗后,患者的CD3⁺、CD4⁺、CD4⁺/CD8⁺较治疗前升高,CD8⁺较治疗前下降（P<0.05）。治疗后,患者的血清白介素-6（IL-6）、C反应蛋白（CRP）和肿瘤坏死因子-α（TNF-α）水平较治疗前下降（P<0.05）。按照ERCP术后是否并发胰腺炎进行分组,其中26例发生胰腺炎的患者纳为并发组,未并发胰腺炎的224例纳为未并发组。单因素分析结果显示,ERCP术后并发胰腺炎与胰管显影、性别、胆总管结石病史、体质量指数（BMI）、Oddi括约肌功能障碍（SOD）病史、ERCP手术时间、多次插管、胰腺炎病史、导丝进入胰管、阻塞性黄疸有关（P<0.05）。多因素Logistic回归分析结果显示：SOD病史、性别为女、BMI≥30 kg/m²、胰腺炎病史、ERCP手术时间≥60 min、多次插管、胰管显影是ERCP术后并发胰腺炎的危险因素（P<0.05）。结论：ERCP治疗老年胆总管结石患者,疗效显著,术中出血量少,患者术后恢复较快,但患者术后有一定的并发症发生,主要以胰腺炎为主。且术后并发胰腺炎的危险因素较多,可采取适当措施改进操作环节,减少术后胰腺炎的发生。