GRP78在食管鳞状细胞癌、不典型增生及正常食管鳞状上皮组织中的表达

目的：检测葡萄糖调节蛋白78（glucose—regulated proteins78,GRP78）在同一食管癌患者食管的鳞状细胞癌、不典型增生及正常鳞状上皮组织中的表达,研究GRP78与食管鳞状细胞癌发生发展的关系。方法：取90例食管鳞状细胞癌患者的病变组织,其病理切片中正常鳞状上皮组织、不典型增生组织和鳞状细胞癌组织均存在,用免疫组化的方法,检测GRP78在3种不同组织中表达的情况,分析G史P78与性别、年龄、浸润深度、分化程度、分期及淋巴结转移等参数之间的关系。结果：GRP78在食管正常鳞状上皮组织、不典型增生组织、鳞状细胞癌组织中的阳性表达率分别为7．8％、85．6％、47．8％,GRP78在正常食管组织、不典型增生组织、食管鳞癌组织3组中表达的差异有显著性意义（P〈0．01）。鳞状细胞癌组织中GRP78表达阴性者,年龄较阳性者大;GRP78表达与肿瘤分化程度、分期、淋巴结转移等有明显相关性。结论：GRP78在食管正常细胞向恶性细胞转化的过程中可能扮演了重要角色,检测GRP78的表达可能有助于对食管鳞状细胞癌的预防及早期诊断。     

GRP78 在食管鳞状细胞癌、不典型增生及正常食管鳞状上皮 组织中的表达

目的:检测葡萄糖调节蛋白78(glucose-regulated proteins 78,GRP78)在同一食管癌患者食管的鳞状细胞癌、不典型增生及正常鳞状上皮组织中的表达,研究GRP78与食管鳞状细胞癌发生发展的关系。方法:取90例食管鳞状细胞癌患者的病变组织,其病理切片中正常鳞状上皮组织、不典型增生组织和鳞状细胞癌组织均存在,用免疫组化的方法 ,检测GRP78在3种不同组织中表达的情况,分析GRP78与性别、年龄、浸润深度、分化程度、分期及淋巴结转移等参数之间的关系。结果:GRP78在食管正常鳞状上皮组织、不典型增生组织、鳞状细胞癌组织中的阳性表达率分别为7.8%、85.6%、47.8%,GRP78在正常食管组织、不典型增生组织、食管鳞癌组织3组中表达的差异有显著性意义(P<0.01)。鳞状细胞癌组织中GRP78表达阴性者,年龄较阳性者大;GRP78表达与肿瘤分化程度、分期、淋巴结转移等有明显相关性。结论:GRP78在食管正常细胞向恶性细胞转化的过程中可能扮演了重要角色,检测GRP78的表达可能有助于对食管鳞状细胞癌的预防及早期诊断。     

Pilot feasibility study of in vivo intraoperative quantitative optical coherence tomography of human brain tissue during glioma resection

This study investigates the feasibility of in vivo quantitative optical coherence tomography (OCT) of human brain tissue during glioma resection surgery in six patients. High‐resolution detection of glioma tissue may allow precise and thorough tumor resection while preserving functional brain areas, and improving overall survival. In this study, in vivo 3D OCT datasets were collected during standard surgical procedure, before and after partial resection of the tumor, both from glioma tissue and normal parenchyma. Subsequently, the attenuation coefficient was extracted from the OCT datasets using an automated and validated algorithm. The cortical measurements yield a mean attenuation coefficient of 3.8 ± 1.2 mm^?1 for normal brain tissue and 3.6 ± 1.1 mm^?1 for glioma tissue. The subcortical measurements yield a mean attenuation coefficient of 5.7 ± 2.1 and 4.5 ± 1.6 mm^?1 for, respectively, normal brain tissue and glioma. Although the results are inconclusive with respect to trends in attenuation coefficient between normal and glioma tissue due to the small sample size, the results are in the range of previously reported values. Therefore, we conclude that the proposed method for quantitative in vivo OCT of human brain tissue is feasible during glioma resection surgery.     

食管鳞状细胞癌中HDGF、VEGF表达与微血管形成的关系

目的:研究食管鳞状细胞癌中肝癌衍生生长因子(HDGF)、血管内皮生长因子(VEGF)的表达及其与微血管形成的关系。方法:通过免疫组化SABC法检测和比较68例食管鳞癌、20例切缘正常组织中HDGF、VEGF的表达和CD34标记的微血管密度(MVD),分析HDGF和VEGF表达之间的关系及其与食管鳞癌患者临床病理因素和食管癌组织MVD值的关系。结果:食管鳞癌组织中HDGF(63.2%)和VEGF(72.1%)的阳性表达率均明显高于切缘正常粘膜组织(15.0%、20.0%)(P0.05),食管鳞癌组织和切缘正常粘膜组织中的MVD值分别为35.48±5.75和13.50±2.1(P0.05)。食管鳞癌组织HDGF的阳性表达率仅与其临床分期明显相关(P0.05),而VEGF的阳性表达率与其淋巴结转移、临床分期均显著相关(P0.05),二者在食管鳞癌组织中的表达呈显著正相关(P0.05)。食管鳞癌组织中HDGF、VEGF阳性表达组MVD值均明显高于HDGF、VEGF阴性表达组(P0.05)。结论:HDGF可能通过诱导VEGF的产生,从而促进血管生成,参与食管鳞癌的发生、发展及转移。     

High-Throughput Genotyping in Metastatic Esophageal Squamous Cell Carcinoma Identifies Phosphoinositide-3-Kinase and BRAF Mutations

Background

Given the high incidence of metastatic esophageal squamous cell carcinoma, especially in Asia, we screened for the presence of somatic mutations using OncoMap platform with the aim of defining subsets of patients who may be potential candidate for targeted therapy.

Methods and Materials

We analyzed 87 tissue specimens obtained from 80 patients who were pathologically confirmed with esophageal squamous cell carcinoma and received 5-fluoropyrimidine/platinum-based chemotherapy. OncoMap 4.0, a mass-spectrometry based assay, was used to interrogate 471 oncogenic mutations in 41 commonly mutated genes. Tumor specimens were prepared from primary cancer sites in 70 patients and from metastatic sites in 17 patients. In order to test the concordance between primary and metastatic sites from the patient for mutations, we analyzed 7 paired (primary-metastatic) specimens. All specimens were formalin-fixed paraffin embedded tissues and tumor content was >70%.

Results

In total, we have detected 20 hotspot mutations out of 80 patients screened. The most frequent mutation was PIK3CA mutation (four E545K, five H1047R and one H1047L) (N = 10, 11.5%) followed by MLH1 V384D (N = 7, 8.0%), TP53 (R306, R175H and R273C) (N = 3, 3.5%), BRAF V600E (N = 1, 1.2%), CTNNB1 D32N (N = 1, 1.2%), and EGFR P733L (N = 1, 1.2%). Distributions of somatic mutations were not different according to anatomic sites of esophageal cancer (cervical/upper, mid, lower). In addition, there was no difference in frequency of mutations between primary-metastasis paired samples.

Conclusions

Our study led to the detection of potentially druggable mutations in esophageal SCC which may guide novel therapies in small subsets of esophageal cancer patients.     

突触融合蛋白6在食管鳞状细胞癌中的表达及临床意义

目的:探讨突触融合蛋白6(syntaxin 6,STX6)蛋白在食管鳞状细胞癌(esophageal squamous carcinoma,ESCC)组织中的表达及临床意义。方法:回顾性分析了241位食管癌病人的临床资料,应用免疫组织化学方法及Western blot法检测肿瘤组织和癌旁食管组织中STX6蛋白的表达情况,并进行统计分析,分析STX6的表达水平与各临床指标间的关系。结果:免疫组织化学法及Western blot结果均显示食管鳞状细胞癌组织中STX6的水平较临近的癌旁食管组织显著上升。食管癌患者病变组织中STX6的表达水平与性别、肿瘤分化程度、有无淋巴结转移以及肿瘤分期显著相关(P0.05),与患者的肿瘤大小、年龄没有明显的相关性(P0.05)。结论:STX6在食管癌的发生发展过程中起着重要的作用,是一个有价值的诊断及治疗靶点。     

NFE2L1在食管鳞癌中的表达及临床意义

目的:通过检测食管鳞癌标本中核因子E2相关因子1(NFE2L1)的表达情况,探究NFE2L1在食管鳞癌发生发展中的作用,为食管鳞癌的诊治以及预后评估提供新的思路。方法:收集我校附院2016-2017年经病理组织学检查诊断为食管鳞癌的手术标本40例及其对应的癌旁组织并从NCBI数据库下载GEO测序数据,应用定量PCR、免疫组化和生物信息分析等方法检测分析NFE2L1基因的表达情况。结果:在食管鳞癌组织中,NFE2L1表达阳性31例(77.5%),癌旁组织阳性表达17例(42.5%),两组比较差异显著有统计学意义(P0.001);进一步发现NFE2L1的阳性表达与肿瘤分化程度和淋巴转移相关(P0.05)。但在不同年龄、性别、浸润深度及不同部位之间差异无统计学意义。GEO数据分析结果显示NFE2L1在食管鳞癌组织显著高表达(P0.01),只是未达到显著差异表达的阈值标准(即变化倍数小于2倍)。结论:NFE2L1在食管鳞癌中高表达,表达的高低与食管鳞癌的发生进展密切相关。     

MRP2 蛋白在食管鳞癌中的表达及其与化疗耐药的相关性

目的:探讨MRP2蛋白在食管鳞癌组织中的表达及其与食管癌化疗耐药的关系。方法:收集原发性食管鳞癌手术标本70例,采用免疫组织化学Envision法检测食管鳞癌组织及其癌旁组织中MRP2蛋白的表达情况,并采用MTT法检测食管鳞癌组织对临床常用化疗药物的敏感性,分析其表达与食管癌化疗耐药的关系。结果:70例食管鳞癌组织及其癌旁正常组织中的阳性表达率分别为58.6%及5.0%。MRP2蛋白在食管鳞癌组织中的阳性表达率明显高于癌旁正常食管组织(P<0.01)。食管鳞癌组织对环磷酰胺、5-氟尿嘧啶、吉西他滨、顺铂、卡铂、阿霉素、长春瑞滨、羟喜树碱等化疗药物的敏感性与其相应癌组织中MRP2表达明显相关(P<0.01)。结论:MRP2的表达与食管鳞癌对多种化疗药物耐药有较好的相关性,推测食管鳞癌组织中MRP2的高表达可能对化疗耐药性的发生发展具有促进作用。     

Multispectral near infrared absorption imaging for histology of skin cancer

Multispectral imaging combines the spectral resolution of spectroscopy with the spatial resolution of imaging and is therefore very useful for biomedical applications. Currently, histological diagnostics use mainly stainings with standard dyes (eg, hematoxylin + eosin) to identify tumors. This method is not applicable in vivo and provides low amounts of chemical information. Biomolecules absorb near infrared light (NIR, 800‐1700 nm) at different wavelengths, which could be used to fingerprint tissue. Here, we built a NIR multispectral absorption imaging setup to study skin tissue samples. NIR light (900‐1500 nm) was used for homogenous wide‐field transmission illumination and detected by a cooled InGaAs camera. In this setup, images I(x, y, λ) from dermatological samples (melanoma, nodular basal‐cell carcinoma, squamous‐cell carcinoma) were acquired to distinguish healthy from diseased tissue regions. In summary, we show the potential of multispectral NIR imaging for cancer diagnostics.      

Increased expression of MARCH8, an E3 ubiquitin ligase,is associated with growth of esophageal tumor

Background

Herein, for the first time, we report aberrant expression of membrane-associated RING-CH8 (MARCH8) in human esophageal squamous cell carcinoma. MARCH8 is a member of the recently discovered MARCH family of really interesting new genes (RING) E3 ligases. Though initial studies primarily focused on its immunomodulatory role, the newly discovered targets of this E3 ligase point towards its possible role in other biological processes such as embryogenesis and inhibition of apoptosis. However, its relevance in cancers is yet to be elucidated.

Methods

We carried out quantitative real time PCR and immunohistochemistry to examine the levels of MARCH8 mRNA and protein in esophageal squamous cell carcinoma tissues. The role of MARCH8 in esophageal cancer cells was evaluated by cell proliferation, clonogenic and migration/invasion assays and flow cytometry with MARCH8 gene knockdown.

Results

Significantly increased expression of MARCH8 mRNA was found in esophageal squamous cell carcinoma as compared to distant matched non-malignant tissues (p = 0.024, AUC = 0.654). Immunohistochemical analysis revealed overexpression of MARCH8 protein in 86% of esophageal squamous cell carcinoma tissues (p < 0.001, AUC = 0.908). Interestingly, intense nuclear staining of MARCH8 protein was detected in cancer cells in addition to its cytoplasmic expression. Knockdown of MARCH8 resulted in decreased proliferation, migration, invasion and clonogenic potential of esophageal cancer cells. In addition to this, silencing of MARCH8 induced apoptosis in esophageal cancer cells which was measured by cell cycle distribution assay which showed increase in sub G0 and G2/M populations (cell death) and decrease in S-phase population. To further check the type of apoptosis induced by MARCH8 silencing, annexin assay was performed which showed significant increase in the number of cells in early apoptotic phase.

Conclusions

Overall, increased expression of MARCH8 gene in preneoplastic and neoplastic esophageal tissues and its knockdown effect on cancer cell properties demonstrated herein points towards the potential role of this protein in esophageal tumorigenesis.

     

MICROSENSOR STUDIES OF OXYGEN AND LIGHT DISTRIBUTION IN THE GREEN MACROALGA CODIUM FRAGILE1

Scalar irradiance, oxygen concentration, and oxygenic photosynthesis were measured at 0.1 mm spatial resolution within the tissue of the siphonous green macroalga Codium fragile subsp. tomentosoides (van Goor) Silva by fiber-optic scalar irradiance microsensors and oxygen microelectrodes. The scalar irradiance of visible light was strongly attenuated in the outer 0.2 mm of the tissue but was nearly constant for the subsequent 1.0 mm of photo-synthetic tissue. Far-red scalar irradiance at 750 nm increased below the tissue surface to a maximum of 200% of incident irradiance at 1.2 mm depth due to multiple scattering in the medullary tissue. The constant intensity of visible light below 0.2 mm was thus a result of the combined effects of absorption and backscattering from the medulla. The oxygen exchange between the alga and the surrounding water was diffusion-limited with a steep O₂gradient inside and around the alga. In darkness, the tissue below 0.6 mm became anoxic, and endophytic extracellular space provided an environment where anoxygenic microbial processes may occur. When illuminated at 160 nmol photons·^?2·^?1, O₂ concentrations exceeded ambient levels throughout the thallus, with a maximum of 250% of air saturation just below the surface. The amplitude of oxygen variation was buffered by gas bubbles formed in the medullary tissue.     

microRNA-21对人舌鳞癌细胞的增殖和凋亡的影响

目的:探讨micro RNA-21(mi R-21)对人舌鳞癌细胞增殖和凋亡的影响。方法:选取8例舌鳞癌组织和4例癌旁组织为研究材料,采用实时荧光定量聚合酶链式反应(q RT-PCR)法对舌鳞癌及癌旁组织中的mi R-21相对表达量进行检测,利用人工合成的mi R-21mimic对人舌鳞癌Tca8113细胞进行瞬时转染,采用q RT-PCR法对Tca8113细胞中mi R-21相对表达量进行检测,采用四唑盐比色法(MTT)法对Tca8113细胞增殖情况进行检测,采用流式细胞术对Tca8113细胞周期与凋亡情况进行检测。结果:舌鳞癌组织中mi R-21的相对表达量(3.502±0.674),高于癌旁组织(0.998±0.192),差异有统计学意义(P0.05)。mi R-21mimic导致了Tca8113细胞中的mi R-21相对表达量上调(6.864±1.324),明显高于对照scramble组[(0.997±0.187),P0.05],对Tca8113细胞的增殖发挥了促进作用(P0.05)。经mi R-21mimic转染之后,Tca8113细胞进入S期的细胞出现了明显的增加[(27.4±5.1)%vs(48.6±8.7)%,P0.05],处于G1期的细胞出现了显著的减少[(56.3±9.6)%vs(36.2±7.2)%,P0.05],细胞凋亡数量出现了显著减少[(9.4±2.3)%vs(18.6±3.9)%,P0.05]。结论:mi R-21在舌鳞癌组织中高表达,过表达mi R-21有效促进了Tca8113细胞的增殖,抑制细胞的凋亡,mi R-21在舌鳞癌诊断和治疗可能具有一定的新型靶点价值。     

A novel microemulsion-based isotonic perfusate modulated by Ringer’s solution for improved microdialysis recovery of liposoluble substances

Background

Microdialysis is promising technique for dynamic microbiochemical sampling from tissues. However, the application of typical aqueous perfusates to liposoluble substances is limited. In this study, a novel microemulsion (ME)-based isotonic perfusate (RS-ME) was prepared to improve the recovery of liposoluble components using microdialysis probes.

Results

Based on pseudo-ternary phase diagrams and comparisons of the ME area, Kolliphor^® EL and Transcutol^® P were selected as the surfactant and co-surfactant, respectively, with a weight ratio (Km) of 2:1 and ethyl oleate as the oil phase. The ME was mixed with Ringer’s solution at a 1:6 ratio (v/v) to obtain the isotonic RS-ME. The droplet size distribution of the ME in RS-ME was 78.3?±?9.2 nm, with a zeta potential of ??3.5?±?0.3 mV. By microdialysis perfusion, RS-ME achieved higher recovery rates of the poorly water-soluble compounds evodiamine (EVO) and ruthenium (RUT), i.e., 58.36?±?0.57% and 49.40?±?0.57%, respectively, than those of 20% (v/v) PEG 400 Ringer's solution (RS-PEG) and 10% (v/v) ethanol Ringer’s solution (RS-EtOH). In vivo microdialysis experiments confirmed that RS-ME captured EVO and RUT molecules around the dialysis membrane more efficiently and exhibited less spreading than RS-PEG and RS-EtOH.

Conclusions

Owing to the nanosized droplets formed by lipid components in the RS-ME and the limited dispersion out of the dialysis membrane, we obtained good biocompatibility and reliable dialysis results, without affecting the tissue microenvironment. As a novel perfusate, RS-ME provides an easy and reliable approach to the microdialysis sampling of fat-soluble components.

     

Wet Chemical Colorimetric Estimation of CO: An Update on the Reduced Silver Sol Spectral and Kinetic Transformations to Silver nanoparticle

Measurements of ppm (v/v) level CO_g concentration is conveniently performed by its preconcentration in alkaline absorber solution of Ag⁺-(4)- HCO₂-C₆H₄-SO₂NH₂ complex, followed by a spectral measurement of the reduced silver sol. In this study, the transitory nature of this latter species and its subsequent real-time transformation to silver nanoparticle are presented. These results were based on spectral measurements made under varying concentrations of alkali, (4)-HCO₂-C₆H₄-SO₂NH₂, and Ag⁺ in the absorber solution, and in the presence of a wide range of sampled CO_g concentration. The initially created light yellow colored sol with its broad absorption profile peaking at 380 nm and absorption coefficient 3500?±?300 cm^?1 M^?1 (related to the amount of sampled [CO_g] as standardized by gas chromatographic analysis) changed into the characteristic yellow orange nanoparticle with its plasmon band peak absorption at 425 nm and absorption coefficient 6350?±?300 cm^?1 M^?1. Under different sampling conditions, the respective first-order conversion rates varied between 0.03 and 0.15 h^?1, whereas simultaneous dynamic light scattering measurements revealed steady growth of the averaged particle size ranging from 60 to 300 nm.     

Controlling the near‐infrared transparency of costal cartilage by impregnation with clearing agents and magnetite nanoparticles

Penetration depth of near‐infrared laser radiation to costal cartilage is controlled by the tissue absorption and scattering, and it is the critical parameter to provide the relaxation of mechanical stress throughout the whole thickness of cartilage implant. To enhance the penetration for the laser radiation on 1.56 μm, the optical clearing solutions of glycerol and fructose of various concentrations are tested. The effective and reversible tissue clearance was achieved. However, the increasing absorption of radiation should be concerned: 5°C‐8°C increase of tissue temperature was detected. Laser parameters used for stress relaxation in cartilage should be optimized when applying optical clearing agents. To concentrate the absorption in the superficial tissue layers, magnetite nanoparticle (NP) dispersions with the mean size 95 ± 5 nm and concentration 3.9 ± 1.1 × 10¹¹ particles/mL are applied. The significant increase in the tissue heating rate was observed along with the decrease in its transparency. Using NPs the respective laser power can be decreased, allowing us to obtain the working temperature locally with reduced thermal effect on the surrounding tissue.      

Sporogenic and vegetative tissues of Saccharina latissima (Laminariales,Phaeophyceae) exhibit distinctive sensitivity to experimentally enhanced ultraviolet radiation : photosynthetically active radiation ratio

Fertile Saccharina latissima sporophytes, collected in the Kongsfjorden, Ny‐Ålesund, Spitsbergen, Norway (78°56.87′ N, 11°51.64′ E) were investigated in relation to its sensitivity to experimentally enhanced ultraviolet radiation : photosynthetically active radiation (UVR : PAR) ratios. Irradiance of UVR were 4.30 W m^?2 of UV‐A (320–400 nm) and 0.40 W m^?2 of UV‐B (280–320 nm), and PAR (400–700 nm) was ～4.30 W m^?2 (=20 µmol photons m^?2 s^?1). Excised soral (sporogenic) and non‐soral (vegetative) tissues were separately irradiated for 16 h at 7°C. Transmission electron microscopy showed abundant occurrence of physodes, electron dense particles (～300–600 nm) in the sorus. Paraphysis cells, with partly crystalline content, large mitochondria and abundant golgi bodies were towering over the sporangia. In soral tissue, cells were not visibly altered by the PAR + UVR irradiation. The chloroplasts, flagella and nucleus of unreleased meiospores inside the sporangial parent cells were visibly intact. Severe changes in the chloroplast structure of vegetative tissue occurred after PAR + UVR irradiation. These changes included wrinkling and dilatation of the thylakoid membranes, and appearance of electron translucent areas inside the chloroplasts. In vegetative cells exposed to PAR + UVR, the total amount of physodes, was slightly higher as in cells exposed to PAR only. Initial values of optimum quantum yield of photosystem II (F_v/F_m) were 0.743 ± 0.04 in non‐soral and 0.633 ± 0.04 in soral tissue. Vegetative tissue was observed to be more sensitive to radiant exposure of PAR and PAR + UVR compared to reproductive tissue. Under PAR, a 20% reduction in Fv/Fm was observed in non‐soral compared to no reduction in soral tissue, whereas under PAR + UVR, 60% and 33% reduction in Fv/Fm was observed in non‐soral and soral tissues, respectively. This can be attributed to the corresponding three times higher antiradical power (ARP) capacity in soral compared to non‐soral tissue.     

Seasonal variation in steroid hormones and blood parameters in cage-farmed European sea bass (Dicentrarchus labrax L.)

For a period of 1 year, some blood parameters were evaluated on a monthly basis in a population of adult European sea bass (Dicentrarchus labrax L.) intensively reared in floating marine cages (Ionian Sea, Mediterranean). From April (13 months old) to July (16 months old) males (35–50%) and sexually indeterminate individuals were collected. From August to March (24 months old) only males were sampled. During this period the percentage of spermiated males was highest (100%) from November (20 months old) to January (22 months old). Plasma testosterone in males was inversely related to sunlight (h month^?1) and was elevated between October and January, when males first achieved sexual maturity. Testosterone showed the highest value (0.49 ± 0.02 ng ml^?1) in January and the lowest (0.09 ± 0.02 ng ml^?1) in March. Haematocrit, red blood cell counts and haemoglobin concentration were elevated from November to March, being inversely related to sunlight. The two latter parameters were also inversely related to daily food intake. Haematocrit, red blood cell counts and haemoglobin concentration were highest in December [53 ± 1%, (5.36 ± 0.06) × 10⁶ mm^?3, 10.08 ± 0.14 g 100 ml^?1, respectively] and lowest in June [35 ± 1%, (3.33 ± 0.05) × 10⁶ mm^?3, 6.47 ± 0.13 g 100 ml^?1, respectively]. White blood cell counts were not correlated with sea water temperature, sunlight or daily food intake. They were highest in February [(8.45 ± 0.20) × 10⁴ mm^?3] and lowest in April [(6.07 ± 0.14) × 10⁴ mm^?3]. Total plasma protein concentration (4.88 ± 0.11–5.93 ± 0.10 g 100 ml^?1) and mean cell volume (93.3 ± 0.9–105.5 ± 1.8 μm³) showed small fluctuations throughout the year. Sexual maturity appears to be a major factor that significantly affects haemopoiesis of D. labrax. This study contributes to the evaluation of normal levels of some blood parameters in European sea bass, which are helpful for the assessment of physiological status and health of this species.