Calcitonin receptor-mediated CFTR activation in human intestinal epithelial cells

High levels of calcitonin (CT) observed in medullary thyroid carcinoma and other CT‐secreting tumours cause severe diarrhoea. Previous studies have suggested that CT induces active chloride secretion. However, the involvement of CT receptor (CTR) and the molecular mechanisms underlying the modulation of intestinal electrolyte secreting intestinal epithelial cells have not been investigated. Therefore, current studies were undertaken to investigate the direct effects of CT on ion transport in intestinal epithelial cells. Real time quantitative RT‐PCR and Western blot analysis demonstrated the expression of CTR in intestinal epithelial T84 cells. Exposure of T84 cells to CT from the basolateral but not from apical side significantly increased short circuit current (I_SC) in a dose‐dependent manner that was blocked by 1 μM of CTR antagonist, CT8–32. CT‐induced I_SC was blocked by replacing chloride in the bath solutions with equimolar gluconate and was significantly inhibited by the specific cystic fibrosis transmembrane conductance regulator (CFTR) inhibitor, CFTR_127inh. Further, biotinylation studies showed that CT increased CFTR levels on the apical membrane. The presence of either the Ca²⁺ chelator, bis(2‐aminophenoxy)ethane tetraacetic acid‐acetoxymethyl (BAPTA‐AM) ester or the protein kinase A (PKA) inhibitor, H89, significantly inhibited I_SC induced by CT (～32–58% reduction). Response to CT was retained after permeabilization of the basolateral or the apical membranes of T84 cells with nystatin. In conclusion, the activation of CTR by CT induced chloride secretion across T84 monolayers via CFTR channel and the involvement of PKA‐ and Ca²⁺‐dependent signalling pathways. These data elucidate the molecular mechanisms underlying CT‐induced diarrhoea.     

伊立替康联合替莫唑胺对儿童难治性实体瘤近期疗效的研究

目的：研究伊立替康（Irinotecan,CPT-11）联合替莫唑胺（Temozolomide,TMZ）对儿童难治性实体瘤的近期疗效;方法：选取了11名难治性儿童实体瘤患者并争取了家属同意,分别给予了伊立替康：60mg／m2／d,ivgtt,连用5天;替莫唑胺：75mg／m2／d,连用5天,在伊立替康前一小时口服;用药期间同时给予思密达或者头孢菌素类抗生素预防药物相关性腹泻,并定期复查血象以及肝肾功;28天一疗程,每疗程行复查CT或MRI进行疗效评定和药物安全性评价。结果：11例患者经过2-6个疗程的治疗：疾病控制率（DCR）达到54％;明显延长了患者的无疾病进展生存期,中位无疾病进展生存期3．3个月。用药期间不良事件主要包括0-Ⅲ级的药物相关性腹泻、呕吐、血小板减少、贫血等;结论：伊立替康联合替莫唑胺方案能明显延长儿童难治实体瘤患者未进展存活时间,且耐受性好、安全性高,可以作为儿童晚期实体瘤患者的首选治疗方案。     

认知负荷对色词Stroop干扰效应的影响

目的:依据负荷理论,认知负荷会导致选择性注意任务中干扰刺激干扰效应的增加。本研究目的在于检验两种类型的认知负荷对色词Stroop干扰效应的影响。方法:实验一通过比较色词Stroop任务与言语工作记忆任务同时进行的双任务区组(高负荷条件)与单任务区组(低负荷条件)来检验认知负荷对构成Stroop干扰效应的语义冲突及反应冲突产生的影响。实验二则通过比较色词Stroop任务与言语工作记忆任务相协调的双任务区组(高负荷条件)与单任务区组(低负荷条件)来操纵认知负荷。结果:实验一的结果显示工作记忆负荷变化对语义冲突及反应冲突均不产生影响。实验二发现虽然在反应时指标上认知负荷与一致性之间未产生显著交互作用,但在错误率指标上两者交互作用达到显著,说明在高认知负荷条件下反应冲突显著增加了。结论:认知负荷对选择性注意任务中干扰刺激干扰效应的影响取决于认知负荷类型及冲突所发生的水平     

伊立替康联合替莫唑胺对儿童难治性实体瘤近期疗效的研究

目的:研究伊立替康(Irinotecan,CPT-11)联合替莫唑胺(Temozolomide,TMZ)对儿童难治性实体瘤的近期疗效;方法:选取了11名难治性儿童实体瘤患者并争取了家属同意,分别给予了伊立替康:60mg/m~2/d,ivgtt,连用5天;替莫唑胺:75mg/m~2/d,连用5天,在伊立替康前一小时口服;用药期间同时给予思密达或者头孢菌素类抗生素预防药物相关性腹泻,并定期复查血象以及肝肾功;28天一疗程,每疗程行复查CT或MRI进行疗效评定和药物安全性评价。结果:11例患者经过2-6个疗程的治疗;疾病控制率(DCR)达到54%;明显延长了患者的无疾病进展生存期,中位无疾病进展生存期3.3个月。用药期间不良事件主要包括0-Ⅲ级的药物相关性腹泻、呕吐、血小板减少、贫血等;结论:伊立替康联合替莫唑胺方案能明显延长儿童难治实体瘤患者未进展存活时间,且耐受性好、安全性高,可以作为儿童晚期实体瘤患者的首选治疗方案。     

Serum levels of angiogenic and anti-angiogenic factors: their prognostic relevance in locally advanced hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is a prototype tumor wherein angiogenesis plays a vital role in its progression. The role of VEGF, a major angiogenic factor in HCC is known; however, the role of anti-angiogenic factors simultaneously with the angiogenic factors has not been studied before. Hence, in this study, the serum levels of major angiogenic [Vascular Endothelial Growth Factor (VEGF), angiopoietin-2 (Ang-2)] and anti-angiogenic (endostatin, angiostatin) factors were analyzed and correlated with clinico-radiological features and with outcome. A total of 150 patients (50 HCC, 50 cirrhosis and 50 chronic hepatitis) and 50 healthy controls were enrolled in this study. Serum levels of VEGF, Ang-2, endostatin, and angiostatin were estimated by enzyme-linked immunosorbent assay. HCC shows significantly elevated serum levels of angiogenic factors VEGF and Ang-2 and of anti-angiogenic factors endostatin and angiostatin. ROC curve analysis for serum VEGF yielded an optimal cut-off value of 225.14 pg/ml, with a sensitivity of 78 % and specificity of 84.7 % for a diagnosis of HCC and its distinction from other group. Using this value, the univariate and multivariate analysis revealed significantly poor outcome in patients with higher levels of serum VEGF (p = 0.009). Combinatorial analysis revealed that patients with higher levels of both angiogenic and anti-angiogenic factors showed poor outcome. Serum VEGF correlates with poor survival of HCC patients and, therefore, serves as a non-invasive biomarker of poor prognosis. Moreover, elevated levels of anti-angiogenic factors occur endogenously in HCC patients.     

Phylogenetic position of Indian termites (Isoptera: Termitidae) with their respective genera inferred from DNA sequence analysis of the mitochondrial cytochrome oxidase gene subunit I compared to subunit II

It has been shown that over-expression of Special AT-rich binding protein 1 (SATB1) in breast cancer predicts a poor prognosis. This study was aimed at investigating the effects of silencing SATB1 on mesenchymal derived human osteosarcoma U2OS cells and the underlying mechanisms. The expressions of SATB1 and the related genes in the cells were detected by qRT-PCR and/or Western Blotting. SATB1 silencing was achieved by stable transfection with the vectors expressing small hairpin RNA versus SATB1. Cell proliferation was detected in a microplate reader with Cell Counting Kit-8 and the cell cycle was analyzed by flow cytometry using a cell cycle detection kit. The study found that SATB1 was particularly over-expressed in human osteosarcoma U2OS. Silencing SATB1 inhibited the proliferation of U2OS. It was found that inhibition of cell proliferation resulted from cell cycle arrest due to down-regulated expression of CFGF and JunB. The over-expression of SATB1 is responsible for abnormal proliferation of mesenchymal derived human Osteosatcoma U2OS cells, indicating that silencing SATB1 expression in the cells might be developed as an efficient osteosarcoma therapy. CTGF and JunB were involved in SATB1-mediated proliferation of U2OS cells.     

T16Ainh-A01对耳蜗血管纹毛细血管内皮细胞凋亡及衰老的影响*

目的: 原代培养豚鼠耳蜗血管纹毛细血管内皮细胞(ECs),探讨跨膜蛋白16A(TMEM16A)在耳蜗血管纹毛细血管ECs衰老过程中的变化及对耳蜗血管纹毛细血管ECs凋亡及衰老的影响。方法: 原代培养耳蜗血管纹毛细血管ECs,细胞传代构建衰老模型并根据CCK-8及β-半乳糖苷酶(SA-β-gal)染色评估细胞衰老程度,衰老细胞被随机分为衰老组(P12)、溶剂组(P12+DMSO)、T16Ainh-A01组(P12+T16Ainh-A01),免疫荧光及Western blot检测TMEM16A在ECs上的表达及分布,流式细胞术检测各组细胞凋亡率,Western blot检测各组Bax、Bcl-2、cleaved casepase-3蛋白表达水平。结果: 原代培养的耳蜗血管纹毛细血管ECs阳性率在95%以上,并确定第12代耳蜗血管纹毛细血管ECs为衰老组,与年轻组ECs相比,衰老组ECs上TMEM16A荧光及蛋白表达显著增强(P＜0.05),细胞凋亡率升高,衰老组给予T16Ainh-A01干预24 h后,Bax、cleaved casepase-3的蛋白表达下调(P＜0.01),Bcl-2的蛋白表达上调(P＜0.05),凋亡率下降且SA-β-gal阳性细胞率明显下降(P＜0.01)。结论: 衰老耳蜗血管纹毛细血管ECs凋亡增多且TMEM16A表达增加,TMEM16A特异性阻断剂T16Ainh-A01可以降低耳蜗血管纹毛细血管ECs的凋亡和衰老程度,提示TMEM16A可能参与耳蜗血管纹毛细血管ECs的凋亡和衰老过程。     

Integrated sedimentological,ichnological and sequence stratigraphical studies of the Koti Dhaman Formation (Tal Group), Nigali Dhar Syncline,Lesser Himalaya,India: paleoenvironmental,paleoecological, paleogeographic significance

Abstract

Integrated ichnology, sedimentology and sequence stratigraphy of the Lower Quartzite Member to the Arkosic Sandstone Member of the Koti Dhaman Formation (Cambrian Series 2, Stage 4), Tal Group, Nigali Dhar Syncline, Lesser Himalayan lithotectonic zone are presented. Trilobite traces of Gondwanan affinity i.e., Cruziana salomonis, Cruziana fasciculata, Rusophycus dispar and Rusophycus burjensis are recorded along with Arenicolites isp. and Skolithos isp. from the Lower Quartzite Member. A rich and diverse ichnoassemblage attributed to the Cruziana ichnofacies is described for the first time from the Arkosic Sandstone Member of the same formation. Seven ichnofossil assemblages, i.e., Cruziana-Rusophycus, Planolites-Palaeophycus, Cruziana problematica, Diplichnites, Cochlichnus anguineus, Bergaueria perata and Psammichnites gigas have been recognized in the Lower Quartzite to Arkosic Sandstone members of the Koti Dhaman Formation. Seven sedimentary facies i.e., sandstone–shale facies (FT1), cross-bedded (trough and planar) sandstone (FT2), bedded sandstone facies (FT3), shale facies (FT4), shale–sandstone facies (FT5), shale-rippled sandstone facies (FT6) and planar and trough cross-laminated sandstone (FT7) and four facies associations FA1-FA4 are identified in the Koti Dhaman Formation. The formation contains shallowing upward parasequences of a tidal flat complex. Overall, two major events are recognized: i) the break in sedimentation between the Lower Quartzite Member and the overlying Shale Member probably related to forced-regressive event and ii) the facies shift from FT6 to FT7 of the Arkosic Sandstone Member represents an erosive transgressive event; the surface is interpreted as wave ravinement surface, which also serves as a sequence boundary. Integrated ichnology, sedimentology and sequence stratigraphic studies indicate that the Lower Quartzite Member was deposited in a shallow subtidal sand sheet complex and tidal flat complex; the Shale Member was deposited in a mud flat setting of a tidal flat complex, and the Arkosic Sandstone Member in a mixed-flat (tidal flat complex) to sand sheet complex front and margin (subtidal sand sheet complex). Overall, the lower to middle part of the Koti Dhaman Formation represents a tide-dominated shallow subtidal–intertidal to mud-flat subenvironments of the tidal flat complex. A palaeogeographic reconstruction of lower Cambrian (516–514?Ma) is presented based on the distribution of trilobite traces from the Lesser Himalaya and the Bikaner–Nagaur area of Peninsular India (eastern Gondwana), Egypt, Jordan, Turkey (western Gondwana) and Canada (Avalonia).     

基于mtDNA COI基因序列的中国柑橘果实蝇的分子鉴定及柑橘大实蝇种群的遗传多样性分析

【目的】探明危害我国柑橘的实蝇种类以及柑橘大实蝇Bactrocera minax不同地理种群和不同寄主种群的遗传多样性。【方法】利用mtDNA COI基因对危害柑橘的果实蝇进行种类鉴定,采用MEGA软件对其中28个地理种群的535头果实蝇COI基因片段(约505 bp)序列进行比对,分析种间及种内遗传距离,构建系统发育树。使用DnaSP软件分析柑橘大实蝇不同地理种群和不同寄主种群的遗传多样性。【结果】从柑橘虫果内共鉴定出4种实蝇,分别为柑橘大实蝇B.minax、桔小实蝇B.dorsalis、蜜柑大实蝇B.tsuneonis和瑞丽果实蝇B.ruiliensis。这4种实蝇的种间遗传距离为0.0264～0.2410,种内遗传距离为0.0000～0.0140,种间与种内遗传距离没有重叠区域。单个柑橘虫果内一般仅有1种实蝇,极个别柑橘果实可同时被两种实蝇危害(4/43);在这些为害柑橘的实蝇种类中,以柑橘大实蝇的个体数量比例最大,占90.70%。柑橘大实蝇地理种群遗传多样性高,28个种群共有17个单倍型。【结论】柑橘大实蝇是所调查地区柑橘实蝇的绝对优势种,其种群遗传分化程度较高,扩散危害风险大。本研究结果对柑橘果实蝇类的监测和防控具有重要意义。