Effect of a Thermal Spring Water on Carbohydrate-Protein Interactions in In-Vitro Models Implicating Normal Human Keratinocytes and Recombinant Lectins

Background: Sugar moiety of macromolecules is today very well known for its implications in many biological recognition mechanisms including cell-cell, extracellular matrix-cell and/or bacteria-cell interactions. In this context lectins, which are carbohydrate-binding proteins displaying a high affinity for sugar groups of other molecules, are of a great importance, notably in immune response involving bacteria, viruses and fungi. As protein-carbohydrate interactions are often mediated by ions such as calcium, zinc or magnesium, we were prompted to study the effect of a thermal spring water (which contains this type of component) on interactions existing between: 1) osidic receptors of human normal keratinocytes and 2) two lectins greatly implicated in the immune response mechanisms (i.e. the dectin-1 and the langerin), and their ligands. Materials and Methods: In a first series of experiments, we studied the effect of increasing concentrations of a thermal spring water on interactions existing between glycosylated molecules and the osidic receptors expressed at the normal human keratinocytes surface. In a second step, and in order to better understand the putative effect of our thermal spring water on the immune response, we analyzed its effect on the interactions existing between the dectin-1 (implicated in the recognition of bacteria, viruses and fungi) and the langerin (expressed by Langerhans cells, the immune cells of the cutaneous tissue), and their ligands in a model using recombinant human lectins and appropriate binding molecules. Results: We showed here that our thermal spring water was able to reinforce interactions between keratinocytes osidic receptors and some of their ligands, in a dose-related manner: From 8% to 55% of increase with 10% to 30% (v/v) of thermal spring water. In the second part of our studies, we also showed that our thermal spring water was able to modulate interactions between dectin-1 and langerin and their ligands through a biphasic effect: Interactions were enhanced by more than 40% and 20% respectively with 10% of thermal spring water, and return to their basal level or lower for higher concentrations. Conclusion: The tested thermal spring water, probably due to its ionic composition, could significantly affect interactions of osidic receptors with their ligands. This property could be of a great interest to help immune system to maintain an appropriate “vigilance state” by using the thermal water at up to a concentration of 10%, and by avoiding any runaway reaction in case of aggression, by using concentrations higher than 10%. .

Quercetin exerts anti-inflammatory effects via inhibiting tumor necrosis factor-α-induced matrix metalloproteinase-9 expression in normal human gastric epithelial cells

BACKGROUND Gastric injury is the most common digestive system disease worldwide and involves inflammation,which can lead to gastric ulcer or gastric cancer(GC).Matrix metallopeptidase-9[MMP-9(gelatinase-B)]plays an important role in inflammation and GC progression.Quercetin and quercetin-rich diets represent potential food supplements and a source of medications for treating gastric injury given their anti-inflammatory activities.However,the effects and mechanisms of action of quercetin on human chronic gastritis and whether quercetin can relieve symptoms remain unclear.AIM To assess whether tumor necrosis factor-α(TNF-α)-induced MMP-9 expression mediates the anti-inflammatory effects of quercetin in normal human gastric mucosal epithelial cells.METHODS The normal human gastric mucosa epithelial cell line GES-1 was used to establish a normal human gastric epithelial cell model of TNF-α-induced MMP-9 protein overexpression to evaluate the antiinflammatory effects of quercetin.The cell counting Kit-8 assay was used to evaluate the effects of varying quercetin doses on cell viability in the normal GES-1 cell line.Cell migration was measured using Transwell assay.The expression of proto-oncogene tyrosine-protein kinase Src(cSrc),phospho(p)-c-Src,extracellular-signal-regulated kinase 2(ERK2),p-ERK1/2,c-Fos,p-c-Fos,nuclear factor kappa B(NF-κB/p65),and p-p65 and the effects of their inhibitors were examined using Western blot analysis and measurement of luciferase activity.p65 expression was detected by immunofluorescence.MMP-9 m RNA and protein levels were measured by quantitative reverse transcription polymerase chain reaction(q RT–PCR)and gelatin zymography,respectively.RESULTS q RT-PCR and gelatin zymography showed that TNF-αinduced MMP-9 m RNA and protein expression in a dose-and time-dependent manner.These effects were reduced by the pretreatment of GES-1 cells with quercetin or a TNF-αantagonist(TNFR inhibitor)in a dose-and timedependent manner.Quercetin and TNF-αantagonists decreased the TNF-α-induced phosphorylation of c-Src,ERK1/2,c-Fos,and p65 in a dose-and time-dependent manner.Quercetin,TNF-αantagonist,PP1,U0126,and tanshinone IIA(TSIIA)reduced TNF-α-induced c-Fos phosphorylation and AP-1–Luciferase(Luc)activity in a dose-and time-dependent manner.Pretreatment with quercetin,TNF-αantagonist,PP1,U0126,or Bay 11-7082 reduced TNF-α-induced p65 phosphorylation and translocation and p65–Luc activity in a dose-and timedependent manner.TNF-αsignificantly increased GES-1 cell migration,and these results were reduced by pretreatment with quercetin or a TNF-αantagonist.CONCLUSION Quercetin significantly downregulates TNF-α-induced MMP-9 expression in GES-1 cells via the TNFR-c-Src–ERK1/2 and c-Fos or NF-κB pathways.

Gene expression analysis of primary normal human hepatocytes infected with human hepatitis B virus

AIM： To find the relationship between hepatitis B virus （HBV） and hepatocytes during the initial state of infection by cDNA microarray. METHODS： Primary normal human hepatocytes （PNHHs） were isolated and infected with HBV. From the PNHHs, RNA was isolated and inverted into complement DNA （cDNA） with Cy3- or Cy5- labeled dUTP for microarray analysis. The labeled cDNA was hybridized with microarray chip, including 4224 cDNAs. From the image of the microarray, expression profiles were produced and some of them were confirmed by RT-PCR, immunoblot analysis, and NF-κB luciferase reporter assay. RESULTS： From the cDNA microarray, we obtained 98 differentially regulated genes. Of the 98 genes, 53 were up regulated and 45 down regulated. Interestingly, in the up regulated genes, we found the TNF signaling pathway-related genes： LT-α, TRAF2, and NIK. By using RT-PCR, we confirmed the up-regulation of these genes in HepG2, HuhT, and Chang liver cells, which were transfected with pHBV1.2x, a plasmid encoding all HBV messages. Moreover, these three genes participated in HBV- mediated NF-κB activation. CONCLUSION： During the initial state of HBV infection, hepatocytes facilitate the activation of NF-κB through up regulation of LT-α, TRAF2, and NIK.

Effect of a Nutrient Mixture on Fanconi Anemia Fibroblast and Normal Human Dermal Fibroblast: A Comparison

Fanconi anemia (FA) is a fatal heterogeneous autosomal recessive disorder, characterized by progressive bone marrow failure, congenital defect and cancer predisposition. Cell culture from FA fibroblast (FAF) displays certain abnormalities as compared to normal human dermal fibroblast (NHDF). This prompted us to investigate the effect of a specific nutrient mixture (NM) containing ascorbic acid, lysine, proline and green tea extract, which has demonstrated a broad spectrum of pharmacological activities, on FAF compared to NHDF. We investigated the in vitro effect of NM on FAF and NHDF cell proliferation by MTT assay, MMPs secretion by zymography, morphology by H&E staining and apoptosis by green caspase assay. FAF (FA-A: PD20, FA-A: PD220) and NHDF were cultured in modified Dulbecco Eagle media. At near confluence, the cells were treated with different concentrations of NM (0, 50, 100, 250, 500 and 1000 μg/ml) in triplicate. The cells were also treated with PMA to induce MMP-9 activity. NM had no effect on FAF cell viability in both cell lines compared to control. In contrast NM exhibited 20% at 50 and 100, 50% at 250, 60% at 500 and 70% toxicity at 1000 μg/ml on NHDF cells. Zymography demonstrated MMP-2 and MMP-9 on PMA stimulation in FAF and NM inhibited the activity of both MMP-2 and MMP-9 in a dose response fashion with total block at 500 μg/ml. In contrast, NHDF exhibited only MMP-2, both active and inactive forms, and NM inhibited their activities in a dose-dependent manner with total block at 1000 μg/ml. H&E staining did not indicate any morphological changes in FAF nor induced apoptosis at higher concentrations, as seen by caspases assay. However, although no morphological changes in NHDF were noted up to NM 100 μg/ml, progressive changes in cell shrinkage, rounding and nuclear condensation, pertaining to apoptosis, were observed at higher concentrations. These changes were consistent with the results from the green caspases apoptosis assay. Our data demonstrate that NM exhibited different responses toward FAF and NHDF. This may in part be due to elevated chromosomal break, deletion and hypersensitivity to cross linking agents, a DNA repair disorder in FAF that is lacking in NHDF.

Con A affinity glycoproteomics of normal human liver tissue

In order to establish the novel high throughput, high efficiency and low cost technological platform for the research of N-glycoproteomics, to resolve the significance of characteristic expression profile of glycoprotein and to find the proteins with biological functional importance, the glycoproteins with high-mannose core and the two antennary types were purified and enriched by the Con A affinity chromatography. Con A affinity protein expression profiles of normal human liver tissue were gener- ated by using SDS-PAGE, two-dimensional electrophoresis (2-DE) followed by fast fluorescence stain- ing based on multiplexed proteomics (MP) technology. 301 visible protein spots on the gel were de- tected and 85 of glycoproteins were further successfully identified via peptide mass fingerprinting (PMF) by a matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF- MS/MS) and annotated to IPI databases. Identified glycoproteins definitely take part in the regulation of cell cycle and metabolic processes. The glycosylation sites were predicted with NetNGlyc 1.0 and NetOGlyc 3.1 software, meanwhile they were classified according to the geneontology methods. The construction of Con A affinity glycoprotein database of normal human liver tissue would contribute to the subsequent research.

Basic Investigations EXPRESSION OF GAP JUNCTION PROTEIN Cx43 IN CULTURED HUMAN NORMAL AND MALIGNANT LUNG CELLS

Gap junctional intercellular communicationexchange of small molecules and ions between contiguous cells through membranous gap junctional channelsis essential for growth control and tissue homecotasis. This work concerns the functional expression of gap junction protein connexin 43 (Cx43) in normal human lung cells and the changes in lung carcinoma cells. By. using Northern blot hybridization analysis and Cx43 immunocytochemical methods, it was otherved that cultured normal human embryonic lung cells expressed a high level of Cx43 in both mRNA and protein levels.The Cx43 immunofluorescence was localized at cell membrane regions corresponding to the location of gap junctions. These normal lung cells were competent of intercellular communication function as detected by Lucifer yellow dye transfer. In contrast to normal celis, Cx43 mRNA and protein was not detectable in the carcinoma PG cell line. These tumor cells were defective of intercellular communication function. These results demonstrate that Cx43 is expressed in normal cultured human embryonic lung cells but not in lung tumor cells. The lack of intercellular communication in the lung tumor cell line correlates with dysfunctional intercellular communication. The suggestive role of Cx as a tumor suppersor gene is discussed.

Fibrous Structure of Connective Tissue of Normal Human Lamina Cribrosa.

Fibrous components and structural morphology of the connective tissue of the lamina cribrosa obtained from 35 normal human autopsy eyes were examined by histochemical staining, transmission electron microscopic and computer-

Determination of optical properties of normal and adenomatous human colon tissues in vitro using integrating sphere techniques

AIM: The purpose of the present study is to compare the optical properties of normal human colon mucosa/submucosa and muscle layer/chorion, and adenomatous human colon mucosa/submucosa and muscle layer/chorion in vitro at 476.5, 488, 496.5, 514.5 and 532 nm. We believe these differences in optical properties should help differential diagnosis of human colon tissues by using optical methods.METHODS: In vitro optical properties were investigated for four kinds of tissues: normal human colon mucosa/submucosa and muscle layer/chorion, and adenomatous human colon mucosa/submucosa and muscle layer/chorion. Tissue samples were taken from 13 human colons (13 adenomatous, 13 normal). From the normal human colons a total of 26 tissue samples, with a mean thickness of 0.40 mm, were used (13 from mucosa/submucosa and 13 from muscle layer/chorion), and from the adenomatous human bladders a total of 26 tissue samples, with a mean thickness of 0.40 mm, were used (13 from mucosa/submucosa and 13 from muscle layer/chorion). The measurements were performed using a double-integratingsphere setup and the optical properties were assessed from these measurements using the adding-doubling method that was considered reliable.RESULTS: The results of measurement showed that there were significant differences in the absorption coefficients and scattering coefficients between normal and adenomatous human colon mucosa/submucosa at the same wavelength,and there were also significant differences in the two optical parameters between both colon muscle layer/chorion at the same wavelength. And there were large differences in the anisotropy factors between both colon mucosa/submucosa at the same wavelength, there were also large differences in the anisotropy factors between both colon muscle layer/chorion at the same wavelength.There were large differences in the value ranges of the absorption coefficients, scattering coefficients and anisotropy factors between both colon mucosa/submucosa,and there were also large differences in these value ranges between both colon muscle layer/chorion. There are the same orders of magnitude in the absorption coefficients for four kinds of colon tissues. The scattering coefficients of these tissues exceed the absorption coefficients by at least two orders of magnitude.CONCLUSION: There were large differences in the three optical parameters between normal and adenomatous human colon mucosa/submucosa at the same laser wavelength, and there were also large differences in these parameters between both colon muscle layer/chorion at the same laser wavelength. Large differences in optical parameters indicate that there were large differences in compositions and structures between both colon mucosa/submucosa, and between both colon muscle layer/chorion.Optical parameters for four kinds of colon tissues are wavelength dependent, and these differences would be useful and helpful in clinical applications of laser and tumors photodynamic therapy (PDT).

A NEW METHOD TO CONSTRUCT A FULL-LENGTH cDNA LIBRARY OF HUMAN NORMAL BLADDER TISSUE

Objective Using template switch mechanism at the 5’ end of mRNA technique (SMART) to construct a full length cDNA library of human normal bladder tissue. Methods The novel procedures used the template switching activity of powerscript reverse transcriptase to synthesize and anchor first strand cDNA in one step. Following reverse transcription, 5 cycles of PCR were performed using a modified oligo(dT) primer and an anchor primer to enrich the full length cDNA population with 1.0 g human normal bladder poly(A) + RNA, then double strand cDNA was synthesized. After digestion with sfiI and size fractionation by CHROMA SPIN 400 columns, double strand cDNA was ligated into λ TripIEx 2 vector and was packaged. We determined the titer of the primary library and the percentage of recombinant clones and finally amplified the library. Results The titer of the cDNA library constructed was 2.1×10 6 pfu·mL -1 , and the amplified cDNA library was 6×10 11 pfu·mL -1 , the percentage of recombination clones was 99%. Conclusion Using SMART technique helps us to construct full length cDNA library with high efficiency and high capacity which lays solid foundation for screening target genes of bladder diseases with probes and antibodies.

Human Rights Protection under the New Normal State of Economy

The new normal state of economy is not only the overall trend of China's current economic development stage, but also the strategic choice of China for the future economic development, the anticipation for the economic development and its outcome under the new strategy. It is of great strategic significance for the long-term development of China's human rights as a whole, yet it makes a short-term challenge to the realization of the rights of vulnerable groups. Under the new normal state of economy, it should be targeted to strengthen the protection of the rights of vulnerable groups, which is very important not only for the overall development of China's human rights causes, but also for China to cross over 'the middle-income trap' and to achieve sustainable development of economy.

基于“两性一度”目标的正常人体解剖学微课开发

正常人体解剖学是学习其他基础医学、临床医学以及中医学等各专业课程的基石和先修课。由于人体结构复杂、相关名词繁多、记忆内容量大等,传统教学模式教学效果不理想。为提高课程的高阶性、创新性和挑战度,分析课程现状和学情,针对正常人体解剖学的重、难点和典型例题配套制作相应的微课,微课具有多系统融合、直观清晰的优势,与课堂教学配合,可提高教学效果,实现知识掌握与能力和素养培养相结合,提高学生自主学习能力与创新能力。

融合改进LM算法及动态时间规整算法的人体动作捕捉研究

人体动作捕捉是指通过技术手段捕捉人体的运动轨迹和姿态信息,广泛应用于娱乐、体育、医疗等领域.然而,现有的动作捕捉技术存在捕捉不准确、计算效率低等问题,影响了其在实时应用场景中的表现.针对这些问题,引入一种骨骼点坐标拟合优化Levenberg-Marquardt算法,并采用粒子群算法对其加以优化.同时采用动态时间规整算法进行人体动作捕捉及评估,以期实现人体动作的实时捕捉.结果显示,该算法对肩部侧平举这一动作的捕捉准确率最高达到了99.23%,明显优于其余对比算法.此外,该算法对深蹲动作的测试时间最短,仅为1.15 s,表明该算法在进行动作捕捉时具有显著的性能优势及很强的实际应用效果,为人体动作捕捉领域提供了新的解决方案.

未来教师如何培养?——人机协同师范教育创新的理论模型与实践进路

随着智能时代的到来,将人工智能与师范教育进行融合成为教育领域的紧迫任务。文章首先分析了师范教育面临的现实挑战,并探讨了师范教育的培养目标、人机协同对师范教育的作用及影响。之后,立足于当前师范教育的目标及需求,文章依托复杂系统理论,构建了人机协同师范教育创新的理论模型。此模型以人机协同为中心,包含教育资源、课程内容、教学方法、评价体系四个维度,以及基础层、教学实践层、组织管理层、政策战略层四个层面,为理解和实施基于人机协同的师范教育创新提供了一个全面、深入的视角。在此基础上,文章进行了案例分析,并结合分析结果提出人机协同师范教育创新的实践进路,回应了“未来教师如何培养”这一核心问题。文章对人机协同师范教育创新的理论与实践探索,可为未来教师的培养提供新视角,并可为人机协同促进师范教育现代化和教育质量提升提供参考。

Targeting tight junctions during epithelial to mesenchymal transition in human pancreatic cancer

Pancreatic cancer continues to be a leading cause of cancer-related death worldwide and there is an urgent need to develop novel diagnostic and therapeutic strategies to reduce the mortality of patients with this disease. In pancreatic cancer, some tight junction proteins, including claudins, are abnormally regulated and therefore are promising molecular targets for diagnosis, prognosis and therapy. Claudin-4 and-18 are overexpressed in human pancreatic cancer and its precursor lesions. Claudin-4 is a high affinity receptor of Clostridium perfringens enterotoxin(CPE). The cytotoxic effects of CPE and monoclonal antibodies against claudin-4 are useful as novel therapeutic tools for pancreatic cancer. Claudin-18 could be a putative marker and therapeutic target with prognostic implications for patients with pancreatic cancer. Claudin-1,-7, tricellulin and marvelD3 are involved in epithelial to mesenchymal transition(EMT) of pancreatic cancer cells and thus might be useful as biomarkers during disease. Protein kinase C is closely related to EMT of pancreatic cancer and regulates tight junctions of normal human pancreatic duct epithelial cells and the cancer cells. This review focuses on the regulation of tight junctions via protein kinase C during EMT in human pancreatic cancer for the purpose of developing new diagnostic and therapeutic modalities for pancreatic cancer.

信息化技术在任务式教学中的应用实践探索--以师范类生物科学专业人体解剖生理学课程为例

社会发展催生高等师范院校新生态,在信息化教育快速发展的情况下,教师可以从课程建设的角度充分利用技术革命的优势,破解原有课堂上的多种问题。其中任务驱动式教学方法结合信息化技术的利用,能够有效提升教学效果,体现在学生的主动参与性、责任心、学生收获和师生关系等多个方面。以师范类生物科学专业人体解剖生理学课程为例,借助信息化技术探索实践新的“任务育人”教学模式,并进行经验总结和反思。

基于OBE理念的“正常人体结构”课程教学实践探索

文章对高职护理专业“正常人体结构”课程存在课程内容繁多、教学形式单一、教学资源有限等问题进行探讨。基于OBE教学理念,从重构课程教学目标、优化课程教学内容、重建课程教学环节、思政教学融入课堂、积极选用新型教材、持续改善育人环境、丰富课程考评体系等方面进行课程教学实践探索,致力于帮助教师提高教学效果和教学质量,增强学生综合素质,不断提高高职“正常人体结构”课程教学成效和医学专业人才培养质量。

基于深度归一化的任意交互物体检测方法研究

交互物体的检测识别是实现人机交互的一项关键技术,针对人机交互过程中交互物体检测范围受限的问题,本文利用深度归一化提高深度图像质量,提出了一种基于图像分割的任意交互物体检测方法。该方法针对操作人员侧向和正向姿态,分别采用基于显著性检测的图像处理和人体姿态引导的区域生长算法分割目标区域,锚定目标物体边框实现物体检测。最后,进行了交互物体检测实验及不同深度区间位置测距和跟随实验。实验结果表明,所提出的物体检测方法能够实现任意交互物体检测,在交互物体检测方面具有广泛适用性;较小深度区间的归一化能够使物体位置误差变小,提高了物体检测距离精度及机器人跟随效果。

医护生“正常人体结构”课程思政的教学策略及实施

专业教育始终需要围绕专业课程进行,传统课堂仍然是专业人才培养的主要场所。在全国教育“大思政”的背景下,专业课教师应坚持以立德树人作为教育教学的根本任务,积极将思政教育贯穿于学科教学全过程中,实现专业课程教学与思政教育有机结合。本文以医护生“正常人体结构”教学为例,探索该课程思政教学策略及实施方法,探寻课程思政教育的内涵与意义,从而为社会培养优秀的医护人才。

高等师范类Human Library特色服务探讨

在国外Human Library是一种十分盛行的阅读方式。2008年Human Library开始在中国的图书馆界获得了解与关注,同时也引发了实践和研究的热潮。然而对于Human Library来说,在师范类高校图书馆及其服务体系中是否适用,却缺乏相应的实践和深入的思考。结合师范类高校的特点,就开展Human Library活动的可操作性以及Human Library在师范类高校教学实践中的作用等方面进行了深入的探讨和研究。

延安市归一化植被指数时空变化及其驱动因素

【目的】植被是陆地生态系统中的主体,研究区域植被覆盖时空变化并量化气候变化和人类活动的相对贡献,可为区域未来的生态环境建设和可持续发展提供科学指导。【方法】基于延安市2001—2020年的MODIS归一化植被指数(NDVI)数据集和同期气象数据,采用Mann-Kendall趋势分析和偏相关分析、残差分析探讨了NDVI对气候变化和人类活动的响应。【结果】①2001—2020年,延安市NDVI年均值为0.69,整体呈明显上升趋势,空间上呈“南部高、北部低”的分布格局。②不同尺度上气温和降水对NDVI的影响存在明显差异。年尺度上,NDVI受降水的影响大于气温;季节尺度上,春季NDVI受到气温和降水的双重影响,夏季降水对NDVI的影响较大,秋、冬季大部分区域NDVI与气温、降水相关性不显著。③延安市NDVI整体呈现改善趋势,明显改善区域面积占比为83.27%。④气候变化和人类活动对植被变化的影响具有双重效应,受气候变化和人类活动共同影响面积占比为91.26%,两者对植被变化的相对贡献率分别为17.81%和82.19%。【结论】延安市植被整体呈向好态势,相较于气候因素,人类活动对NDVI变化的驱动作用更强。