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Clinical Significance of Subcellular Localization of Maspin in Breast Carcinoma: An Immunohistochemical Study Using Two Different Antibodies. 乳腺癌中Maspin亚细胞定位的临床意义:两种不同抗体的免疫组织化学研究。
IF 1 4区 医学 Q4 MEDICINE, RESEARCH & EXPERIMENTAL Pub Date : 2023-02-01 DOI: 10.33160/yam.2023.02.003
Makoto Wakahara, Keiko Hosoya, Hiroshi Ishii, Yoshihisa Umekita

Background: Maspin is known to be a tumor suppressor protein: however, its prognostic value in patients with breast cancer remains controversial. The key influential factors contributing to this complexity may be the differences in antibodies used, as well as the positive criteria and sample size. To date, no study has investigated the prognostic significance of maspin expression by using two different antibodies in the same cohort. We aimed to clarify whether differences in antibodies could influence on the prognostic value of maspin in breast cancer patients.

Methods: Immunohistochemical analyses using an anti-maspin antibody (clone G167-70) were performed on 164 resected specimens of invasive carcinoma of no special type (NOS). The correlation with clinicopathological factors was compared to previous results using clone EAW24, with longer follow-up duration.

Results: The subcellular localization of maspin expression was as follows: cytoplasmic-only staining, 3 cases (1.8%), pancellular staining, 43 cases (26.2%); and no staining, 118 cases (72.0%). No nuclear-only staining was observed. There was no significant correlation between clinicopathological characteristics and the pancellualr expression of maspin. The pancellular expression group showed a significantly longer disease-free survival (DFS) than the other groups (P = 0.046). When clone EAW24 was used, the cytoplasmic-only staining group showed significantly shorter DFS than the pancellular staining group (P = 0.003).

Conclusion: Clone EAW24 may be superior to clone G167-70 in selecting breast carcinoma with an aggressive phenotype, while clone G167-70 may be superior to clone EAW24 in selecting non-aggressive breast carcinoma.

背景:Maspin是一种已知的肿瘤抑制蛋白,然而,其在乳腺癌患者中的预后价值仍存在争议。造成这种复杂性的关键影响因素可能是所使用抗体的差异,以及阳性标准和样本量。迄今为止,还没有研究通过在同一队列中使用两种不同的抗体来研究maspin表达的预后意义。我们的目的是澄清抗体的差异是否会影响乳腺癌患者的预后价值。方法:采用抗maspin抗体(克隆G167-70)对164例无特殊类型浸润性癌(NOS)切除标本进行免疫组化分析。与临床病理因素的相关性进行比较,使用克隆EAW24,随访时间更长。结果:maspin表达的亚细胞定位:仅细胞质染色3例(1.8%),泛细胞染色43例(26.2%);未染色118例(72.0%)。未见纯核染色。maspin的临床病理特征与全细胞表达无明显相关性。泛细胞表达组的无病生存期(DFS)明显长于其他各组(P = 0.046)。克隆EAW24时,细胞质单染色组DFS明显短于泛细胞染色组(P = 0.003)。结论:克隆EAW24在筛选侵袭性乳腺癌方面可能优于克隆G167-70,而克隆G167-70在筛选非侵袭性乳腺癌方面可能优于克隆EAW24。
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引用次数: 0
Development of a Prognostic Scoring System using MYC Expression and Soluble Interleukin Receptor -2 level for Diffuse Large B-cell Lymphoma. 弥漫性大b细胞淋巴瘤MYC表达和可溶性白细胞介素受体-2水平预后评分系统的建立
IF 1 4区 医学 Q4 MEDICINE, RESEARCH & EXPERIMENTAL Pub Date : 2023-02-01 DOI: 10.33160/yam.2023.02.007
Sayaka Suzuki, Satoshi Kuwamoto, Koji Kawamura, Michiko Matsushita, Toru Motokura, Yuzuru Hosoda, Masaya Maegaki, Rina Hosoda, Kentaro Hara, Yoshihisa Umekita, Tetsuya Fukuda

Background: Diffuse large B-cell lymphoma, not otherwise specified (DLBCL-NOS), is the most frequent type of lymphoid neoplasm.

Methods: We investigated the relationships between clinical factors of DLBCL-NOS and MYC immunohistochemistry (IHC) staining.

Results: A total of 110 patients diagnosed with DLBCL-NOS from 2012 to 2020 at Tottori University Hospital and treated with rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisolone (R-CHOP) chemotherapy were included. IHC staining of MYC in formalin-fixed, paraffin-embedded tumor specimens was performed, and ROC-curve analysis revealed the cut-off value of the MYC positive rate as 55%. The 2-year overall survival (OS) rates of the MYC-negative and -positive groups were 84.7% vs 57.7% (P = 0.0091), and the progression-free survival rates were 77.8% vs 54.7% (P = 0.016), respectively. Multivariate analysis for OS showed prognostic significance of MYC positivity [hazards ratio (HR): 2.496; P = 0.032], and serum levels of soluble interleukin-2 receptor (sIL-2R) > 2000 U/mL (HR: 3.950; P = 0.0019), as well as age > 75 (HR: 2.356; P = 0.068). The original scoring system was developed based on these findings. By assigning one point to each item, age (> 75), MYC positivity, and sIL-2R level (> 2000), all patients were classified into three risk categories: group 1 (0 points), group 2 (1 point), and group 3 (2-3 points). The 2-year survival rates were 100%, 83.0%, and 47.1% for the groups 1, 2, and 3, respectively (P < 0.0001).

Conclusion: We suggest that a prognostic scoring system using MYC expression and soluble interleukin receptor -2 level is useful for the prediction of prognosis, contributing to further stratification in DLBCL-NOS.

背景:弥漫性大b细胞淋巴瘤(DLBCL-NOS)是最常见的淋巴样肿瘤类型。方法:探讨DLBCL-NOS临床因素与MYC免疫组化(IHC)染色的关系。结果:2012 - 2020年在鸟取大学医院诊断为DLBCL-NOS并接受利妥昔单抗、环磷酰胺、阿霉素、长春新碱和强的松龙(R-CHOP)化疗的患者共110例。对福尔马林固定、石蜡包埋的肿瘤标本进行MYC免疫组化染色,roc曲线分析显示MYC阳性率临界值为55%。myc阴性组和阳性组的2年总生存率(OS)分别为84.7%和57.7% (P = 0.0091),无进展生存率分别为77.8%和54.7% (P = 0.016)。多因素分析显示MYC阳性对OS有预后意义[危险比(HR): 2.496;P = 0.032],血清可溶性白细胞介素-2受体(sIL-2R)水平> 2000 U/mL (HR: 3.950;P = 0.0019),年龄> 75岁(HR: 2.356;P = 0.068)。最初的评分系统是基于这些发现开发的。根据年龄(> 75)、MYC阳性、sIL-2R水平(> 2000)各1分,将患者分为1组(0分)、2组(1分)、3组(2-3分)。1、2、3组2年生存率分别为100%、83.0%、47.1% (P < 0.0001)。结论:基于MYC表达和可溶性白细胞介素受体-2水平的预后评分系统有助于预测预后,有助于DLBCL-NOS的进一步分层。
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引用次数: 0
Imaging of Liver Tissues Using a Combination of Silver Impregnation and Low-Vacuum Scanning Electron Microscopy; A Simple Method for the High-Resolution Visualization of Reticulin Structures with Applicability to a Quantitative Analysis. 银浸渍与低真空扫描电镜联合成像肝脏组织一种适用于定量分析的网状结构高分辨率可视化简单方法。
IF 1 4区 医学 Q4 MEDICINE, RESEARCH & EXPERIMENTAL Pub Date : 2023-02-01 DOI: 10.33160/yam.2023.02.009
Mikihiro Izuta, Satoshi Kuwamoto, Tetsutaro Kamiya, Keiko Nagata, Michiko Matsushita, Sumire Inaga, Toshiyuki Kaido, Yoshihisa Umekita, Kazuhiko Hayashi, Yukisato Kitamura

Background: Low-vacuum scanning electron microscopy (LVSEM) enables the detailed three-dimensional imaging of archival tissues without special pretreatments. The clinical utility of LVSEM in the assessment of liver diseases has not yet been defined. So, we investigated the utility of LVSEM imaging in morphological assessments of normal and diseased liver tissues, with a focus on reticulin structures.

Methods: Formalin-fixed tissue samples of two normal livers and two hepatocellular carcinomas with background regenerative nodules/areas were stained with platinum blue stain or silver-impregnated using Watanabe's method and then comparatively observed under LVSEM. We also evaluated the applicability of LVSEM imaging of liver tissues to a quantitative analysis using a digital image analysis technique.

Results: Optimal high-resolution images of reticulin structures were obtained using 10-μm-thick silver-impregnated sections. Reticulin fibers were clearly observed to run dendritically around sinusoids in normal livers, and markedly increased in regenerative nodules/areas. Normal reticulin frameworks were lost in hepatocellular carcinoma, leaving a few fragments of reticulin fibers within tumors. Moreover, when a quantitative analysis was applied to these images, we successfully demonstrated a significantly higher reticulin fiber density in regenerative nodules/areas than in the normal liver (P < 0.05).

Conclusion: We not only obtained detailed three-dimensional images of reticulin structures in various liver tissues by LVSEM combined with silver impregnation but also showed their applicability to a quantitative analysis. The method presented herein may be applied to future studies for the more accurate diagnosis and better classification/risk stratification of various liver diseases.

背景:低真空扫描电子显微镜(LVSEM)无需特殊预处理即可对档案组织进行详细的三维成像。LVSEM在肝脏疾病评估中的临床应用尚未明确。因此,我们研究了LVSEM成像在正常和病变肝脏组织形态学评估中的应用,重点是网状蛋白结构。方法:采用铂蓝染色或Watanabe法浸渍银,分别对2例正常肝脏和2例背景有再生结节/区的肝细胞癌进行福尔马林固定组织,并在LVSEM下进行对比观察。我们还评估了肝组织LVSEM成像在数字图像分析技术定量分析中的适用性。结果:采用10 μm厚的镀银切片,获得了最佳的高分辨率网状蛋白结构图像。在正常肝脏中清晰地观察到网状纤维在窦周围呈树突状分布,在再生结节/区域中明显增加。正常的网状蛋白框架在肝细胞癌中丢失,在肿瘤中留下少量的网状蛋白纤维碎片。此外,当对这些图像进行定量分析时,我们成功地证明了再生结节/区域的网状纤维密度明显高于正常肝脏(P < 0.05)。结论:利用LVSEM结合银浸渍技术,不仅获得了肝脏各组织网状蛋白结构的详细三维图像,而且具有定量分析的适用性。该方法可应用于今后的研究,对各种肝脏疾病进行更准确的诊断和更好的分类/风险分层。
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引用次数: 0
Medical Doctors' Work-Life Balance and the Use of Household Chore Support Services. 医生工作与生活的平衡与家务辅助服务的使用。
IF 1 4区 医学 Q4 MEDICINE, RESEARCH & EXPERIMENTAL Pub Date : 2023-02-01 DOI: 10.33160/yam.2023.02.005
Toshiki Fukuzaki, Sawako Ooba, Kenji Yamaguchi, Ryuko Matsuda, Noriyuki Namba

Background: Achieving an optimal work-life balance (WLB) is an important social issue not only for workers in general but also for doctors due to the shortage of women doctors. The present study aims to survey doctors' WLB and their use of household chore support services (HCSS).

Methods: A questionnaire survey was conducted with doctors working in Tottori Prefecture and a total of 289 responses (212 men, 77 women) were obtained and analyzed. To examine the relationship between gender and satisfaction with working patterns or with life for WLB, as well as the use of HCSS, a chi-squared test was conducted. Furthermore, a chi-squared test was conducted including age, marital status, whether or not they have preschool-aged children, and type of institution the participant worked.

Results: A significant difference was found in type of institution the participant worked regarding satisfaction with work patterns and life. The proportion of those who have used HCSS was 12.5% of the total sample and was significantly higher for women than men (men: 8.5%; women: 23.4%). Regarding the reason to use HCSS, "to reduce the burden of household chores and childcare" and "to secure time for work" were most common with the same proportions, and amongst women, "to secure time for work" was the highest.

Conclusion: Doctors working at a university hospital could have different work patterns and satisfaction with life compared to those working at other types of medical institutions. Additionally, the demand for HCSS was particularly high among women doctors, which suggests that HCSS may be used to reduce the burden of household chores and childcare, as well as to create time for work.

背景:实现最佳的工作与生活平衡(WLB)是一个重要的社会问题,不仅对一般工人,而且由于女医生的短缺,医生。本研究旨在调查医生的工作幸福感与家务支持服务的使用情况。方法:对鸟取县执业医师进行问卷调查,共获得289份问卷,其中男性212份,女性77份。为了检验性别与工作模式或生活满意度之间的关系,以及HCSS的使用,进行了卡方检验。此外,卡方检验包括年龄,婚姻状况,是否有学龄前儿童,以及参与者工作的机构类型。结果:被试在工作型态与生活型态的满意度上,单位类型有显著差异。使用过HCSS的人占总样本的12.5%,女性的比例明显高于男性(男性:8.5%;女性:23.4%)。至于使用家庭福利金的原因,最常见的是“减轻家务和照顾孩子的负担”和“确保有时间工作”,比例相同,而在女性中,“确保有时间工作”的比例最高。结论:高校医院医生的工作模式和生活满意度与其他类型医疗机构不同。此外,女医生对HCSS的需求特别高,这表明HCSS可以用来减轻家务和照顾孩子的负担,并为工作创造时间。
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引用次数: 0
Advantages and Disadvantages of Online Learning Counseling Practice. 在线学习辅导实践的利弊。
IF 1 4区 医学 Q4 MEDICINE, RESEARCH & EXPERIMENTAL Pub Date : 2023-02-01 DOI: 10.33160/yam.2023.02.019
Naomi Sunami

The School of Medicine offers study counseling. In particular, the School of Medicine has many required courses, and it is important to respond to students with learning difficulties. It is necessary to discuss a new approach to learning counseling for such students, considering the prevention of COVID-19 infection, but online learning counseling has not been discussed so far. In the future, flexible selection and response based on an understanding of the advantages and disadvantages of online learning counseling will be required. To overcome this challenge, this paper examines the practice of online learning counseling at the University's School of Medicine in the Corona Disaster. The practice could be characterized by seven advantages [behavioral understanding, physical condition consideration, continuity, confidentiality, sharing, immediacy, and convenience] and two challenges [grasping reactions and interruptions]. Subsequently, it was presented new possibilities for online learning counseling by examining the content related to learning.

医学院提供学习辅导。特别是医学院有很多必修课程,对有学习困难的学生做出回应是很重要的。考虑到COVID-19感染的预防,有必要讨论新的学习辅导方法,但到目前为止还没有讨论在线学习辅导。在未来,需要在了解在线学习咨询的优缺点的基础上灵活选择和应对。为了克服这一挑战,本文研究了在冠状病毒灾难中大学医学院在线学习咨询的实践。该实践具有七个优势(行为理解、身体状况考虑、连续性、保密性、共享性、即时性和便利性)和两个挑战(把握反应和中断)。随后,通过研究与学习相关的内容,提出了在线学习咨询的新可能性。
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引用次数: 1
Bystanders' Willingness to Perform Basic Life Support and Its Relationship with Facilitative and Obstructive Factors: A Nationwide Survey in Japan. 旁观者实施基本生命支持的意愿及其与促进因素和阻碍因素的关系:日本的一项全国性调查。
IF 1 4区 医学 Q4 MEDICINE, RESEARCH & EXPERIMENTAL Pub Date : 2023-02-01 DOI: 10.33160/yam.2023.02.008
Yoshiyuki Hasegawa, Keiichi Hanaki

Background: The administration of basic life support (BLS) by bystanders is essential to improve the survival rates of patients who have experienced out-of-hospital cardiac arrest (OHCA). Although providing BLS to individuals who experience OHCA greatly improves their chances of survival, the actual implementation rate is low. Therefore, we investigated the association between bystanders' willingness to perform BLS and facilitative/obstructive factors with the objective of identifying educational methods that would improve the likelihood of bystanders performing BLS should they encounter a patient with OHCA.

Methods: The study participants included 502 male and 498 female Japanese residents (total, 1000 participants) with no experience in performing BLS and 42 male and 59 female Japanese residents (total 101 participants) with experience in performing BLS. The participants were aged 15-65 years. Both groups graded the strength of their willingness to perform BLS in the future on a 4-point scale, as well as their level of agreement with factors facilitating or obstructing their willingness to perform BLS. These factors were established based on the theory of helping behavior, which defines psychological states when helping others in social psychology.We then analyzed the associations between willingness to perform BLS in the future and their level of agreement with factors facilitating or obstructing their willingness to perform BLS.

Results: The willingness to perform BLS decreased in accordance with the increase in the level of intervention required for patients who experienced OHCA , and was significantly associated with four facilitating factors: sufficient ability and experience to perform BLS, personal advantage, high personal norms, and psychological closeness to the patient.

Conclusion: Our results suggested that workshops and other educational activities focused on these facilitative factors may be helpful in increasing the rate at which bystanders perform BLS.

背景:旁观者给予基本生命支持(BLS)对于提高院外心脏骤停(OHCA)患者的生存率至关重要。虽然向经历过OHCA的个人提供劳工统计局大大提高了他们的生存机会,但实际执行率很低。因此,我们调查了旁观者执行BLS的意愿与促进/阻碍因素之间的关系,目的是确定教育方法,以提高旁观者在遇到OHCA患者时执行BLS的可能性。方法:研究对象为无劳工统计局经验的日本居民502名男性和498名女性(共1000人),有劳工统计局经验的日本居民42名男性和59名女性(共101人)。参与者年龄在15-65岁之间。两组都以4分制对他们未来执行劳工统计局的意愿强度进行评分,以及他们对促进或阻碍他们执行劳工统计局意愿的因素的同意程度。这些因素是在帮助行为理论的基础上建立起来的,帮助行为理论在社会心理学中定义了帮助他人时的心理状态。然后,我们分析了未来执行劳工统计局的意愿与他们对促进或阻碍他们执行劳工统计局意愿的因素的同意程度之间的关系。结果:OHCA患者执行BLS的意愿随干预水平的提高而降低,且与执行BLS的能力和经验充足、个人优势、个人规范高、与患者的心理亲密度四个促进因素显著相关。结论:我们的研究结果表明,关注这些促进因素的研讨会和其他教育活动可能有助于提高旁观者执行劳工统计局的比率。
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引用次数: 0
Phase Lag Analysis Scalp Electroencephalography May Predict Seizure Frequencies in Patients with Childhood Epilepsy with Centrotemporal Spikes. 相位滞后分析头皮脑电图可以预测儿童癫痫患者的发作频率与中央颞叶尖峰。
IF 1 4区 医学 Q4 MEDICINE, RESEARCH & EXPERIMENTAL Pub Date : 2023-02-01 DOI: 10.33160/yam.2023.02.006
Masayoshi Oguri, Tetsuya Okazaki, Tohru Okanishi, Masashi Nishiyama, Sotaro Kanai, Hiroyuki Yamada, Kaoru Ogo, Takashi Himoto, Yoshihiro Maegaki, Ayataka Fujimoto

Background: Childhood epilepsy with centrotemporal spikes (CECTS) is the most common epilepsy syndrome in school-aged children. However, predictors for seizure frequency are yet to be clarified using the phase lag index (PLI) analyses. We investigated PLI of scalp electroencephalography data at onset to identify potential predictive markers for seizure times.

Methods: We compared the PLIs of 13 patients with CECTS and 13 age- and sex-matched healthy controls. For the PLI analysis, we used resting-state electroencephalography data (excluding paroxysmal discharges), and analyzed the mean PLIs among all electrodes and between interest electrodes (C3, C4, P3, P4, T3, and T4) and other electrodes. Furthermore, we compared PLIs between CECTS and control data and analyzed the associations between PLIs and total seizure times in CECTS patients.

Results: No differences were detected in clinical profiles or visual electroencephalography examinations between patients with CECTS and control participants. In patients with CECTS, the mean PLIs among all electrodes and toward interest electrodes were higher at the theta and alpha bands and lower at the delta and gamma bands than those in control participants. Additionally, the mean PLIs toward interest electrodes in the beta frequency band were negatively associated with seizure times (P = 0.02).

Conclusion: The resting-state delta, theta, alpha, and gamma band PLIs might reflect an aberrant brain network in patients with CECTS. The resting-state PLI among the selected electrodes of interest in the beta frequency band may be a predictive marker of seizure times in patients with CECTS.

背景:儿童期癫痫伴中央颞峰(CECTS)是学龄儿童中最常见的癫痫综合征。然而,癫痫发作频率的预测因子尚未通过相位滞后指数(PLI)分析得到明确。我们研究了发病时头皮脑电图数据的PLI,以确定癫痫发作时间的潜在预测标记。方法:我们比较了13例cect患者和13例年龄和性别匹配的健康对照者的PLIs。对于PLI分析,我们使用静息状态脑电图数据(不包括阵发性放电),并分析所有电极之间以及感兴趣电极(C3, C4, P3, P4, T3和T4)与其他电极之间的平均PLI。此外,我们比较了cect患者与对照组的pli数据,并分析了cect患者pli与总发作时间之间的关系。结果:在cect患者和对照组之间的临床资料或视觉脑电图检查没有发现差异。在cect患者中,所有电极和感兴趣电极的平均PLIs在θ和α波段较高,在δ和γ波段较低。此外,在β频带中,朝向感兴趣电极的平均pli与癫痫发作次数呈负相关(P = 0.02)。结论:静息状态δ、θ、α和γ波段PLIs可能反映了cect患者的异常脑网络。在β频带中选择的感兴趣电极的静息状态PLI可能是cect患者癫痫发作时间的预测标记。
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引用次数: 0
Cisplatin Induces Cell Death in Rat Adult Kidney Stem/ Progenitor Cell-Derived Kidney Organoids. 顺铂诱导大鼠成体肾干/祖细胞衍生肾类器官细胞死亡
IF 1 4区 医学 Q4 MEDICINE, RESEARCH & EXPERIMENTAL Pub Date : 2023-02-01 DOI: 10.33160/yam.2023.02.018
Yasushi Kuromi, Shinji Kitamura, Takeshi Endo, Shota Ueno, Kenji Kokura, Tetsuya Ohbayashi

Background: The use of stem/ progenitor cell-derived organoids to evaluate the toxicity of chemical substances has widely increased. Organoids with nephron-like structures (NLS) can be derived from rat adult kidney stem/ progenitor cells (rKS cells) using three-dimensional culture. In this study, we examined the effects of cisplatin, an anticancer drug that induces nephrotoxicity in vivo, on rKS cell-derived NLS.

Methods: Twelve organoids were simultaneously derived from three-dimensionally cultured rKS cells in Matrigel matrices. The surface area of each organoid was measured using microscopy-based imaging, and the morphological changes of NLS were monitored using an image analysis method. NLS were exposed to cisplatin, and their associated effects were assessed.

Results: NLS elongated over time. The surface areas of the 12 organoids were almost constant. Cisplatin exposure induced cell death in NLS and inhibited the increase in the surface area of the organoids.

Conclusion: Cisplatin exposure induces damage to NLS derived from rKS cells. Thus, the organoids may be valuable as an in vitro model to assess nephrotoxicity.

背景:利用干细胞/祖细胞衍生的类器官来评估化学物质的毒性已经广泛增加。具有肾元样结构的类器官(NLS)可以通过三维培养从大鼠成体肾干/祖细胞(rKS细胞)中获得。在这项研究中,我们研究了顺铂(一种体内诱导肾毒性的抗癌药物)对rKS细胞源性NLS的影响。方法:在Matrigel基质中培养rKS细胞,同时获得12个类器官。采用显微成像技术测量各类器官的表面积,采用图像分析方法监测NLS的形态学变化。NLS暴露于顺铂,并评估其相关效应。结果:NLS随时间延长。12个类器官的表面积几乎是恒定的。顺铂暴露诱导NLS细胞死亡,抑制类器官表面积的增加。结论:顺铂暴露可引起rKS细胞NLS损伤。因此,类器官可能是一个有价值的体外模型,以评估肾毒性。
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引用次数: 0
Micall2 Is Responsible for the Malignancy of Clear Cell Renal Cell Carcinoma. Micall2与透明细胞肾细胞癌的恶性相关。
IF 1 4区 医学 Q4 MEDICINE, RESEARCH & EXPERIMENTAL Pub Date : 2023-02-01 DOI: 10.33160/yam.2023.02.021
Xianyou Zeng, Hongquan Wang, Jia Yang, Jing Hu

Background: There lacks a sufficient research on the tumorigenesis of clear cell renal cell carcinoma (ccRCC), causing that the prognosis of ccRCC was not effectively improved. Micall2 contributes to the malignancy of cancer. Moreover, Micall2 is considered a typical cell mobility-promoting factor. However, the relationship between Micall2 and ccRCC malignancy is unknown.

Methods: In this study, we first investigated the expression patterns of Micall2 in ccRCC tissues and cell lines. Next, we explored the in vitro and in vivo roles of Micall2 in ccRCC tumorigenesis based on ccRCC cell lines with different Micall2 expression and gene manipulation assays.

Results: Our study showed that ccRCC tissues and cell lines expressed higher level of Micall2 than paracancerous tissues and normal renal tubular epithelial cell, and Micall2 expression was overexpressed on cancerous tissue with significant metastasis and enlargement. Among three ccRCC cell lines, the expression of Micall2 was the highest in 786-O cells and the lowest in CAKI-1 cells. Moreover, 786-O cells showed the highest malignancy in vitro and in vivo (including proliferation, migration, invasion, reduced E-cadherin expression and tumorigenicity of nude mice in vivo), while CAKI-1 cells showed the contrary results. Furthermore, the upregulated Micall2 by Gene overexpression promoted the proliferation, migration and invasion of ccRCC cells while the downregulated Micall2 by Gene silencing showed the opposite effect.

Conclusion: Micall2, as a pro-tumorigenic gene marker of ccRCC, contributes the malignancy of ccRCC.

背景:透明细胞肾细胞癌(clear cell renal cell carcinoma, ccRCC)的发生机制缺乏足够的研究,导致ccRCC的预后没有得到有效改善。Micall2有助于癌症的恶性化。此外,Micall2被认为是一种典型的细胞移动促进因子。然而,Micall2与ccRCC恶性肿瘤的关系尚不清楚。方法:在本研究中,我们首先研究Micall2在ccRCC组织和细胞系中的表达模式。接下来,我们基于不同Micall2表达和基因操作的ccRCC细胞系,探讨Micall2在ccRCC肿瘤发生中的体外和体内作用。结果:我们的研究表明,ccRCC组织和细胞系Micall2的表达水平高于癌旁组织和正常肾小管上皮细胞,Micall2在转移和扩大明显的癌组织中过表达。在3个ccRCC细胞系中,Micall2在786-O细胞中表达量最高,在CAKI-1细胞中表达量最低。此外,786-O细胞在体外和体内表现出最高的恶性肿瘤(包括增殖、迁移、侵袭、E-cadherin表达降低和裸鼠体内致瘤性),而CAKI-1细胞表现出相反的结果。此外,基因过表达上调Micall2可促进ccRCC细胞的增殖、迁移和侵袭,而基因沉默下调Micall2则相反。结论:Micall2作为ccRCC的致瘤基因标志物,参与了ccRCC的恶性发展。
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引用次数: 1
Pathological Comparison of TDP-43 Between Motor Neurons and Interneurons Expressed by a Tetracycline Repressor System on the Mouse Artificial Chromosome. 四环素抑制系统在小鼠人工染色体上表达运动神经元与中间神经元TDP-43的病理比较
IF 1 4区 医学 Q4 MEDICINE, RESEARCH & EXPERIMENTAL Pub Date : 2023-02-01 DOI: 10.33160/yam.2023.02.004
Shota Togai, Shusei Hamamichi, Yasuhiro Kazuki, Masaharu Hiratsuka

Background: Cytoplasmic mislocalization of TAR-DNA binding protein of 43 kDa (TDP-43) is a major hallmark of amyotrophic lateral sclerosis (ALS). TDP-43 aggregation is detected in the cortical and spinal motor neurons in most ALS cases; however, pathological mechanism of this mislocalized TDP-43 remains unknown.

Methods: We generated a tetracycline-inducible TDP-43 A315T system on a mouse artificial chromosome (MAC) vector to avoid transgene-insertional mutagenesis, established a mouse embryonic stem (ES) cell line holding this MAC vector system, and investigated whether overexpressed exogenous TDP-43 A315T was mislocalized in the cytoplasm of the ES cell-derived neurons and triggered the neurotoxic effects on these cells.

Results: Inducible TDP-43 A315T system was successfully loaded onto the MAC and introduced into the mouse ES cells. These ES cells could differentiate into motor neurons and interneurons. Overexpression of TDP-43 A315T by addition of doxycycline in both neurons resulted in mislocalization to cytoplasm. Mislocalized TDP-43 caused cell death of motor neurons, but not interneurons.

Conclusion: Vulnerability to cytoplasmic mislocalized TDP-43 is selective on neuronal types, whereas mislocalization of overexpressed TDP-43 occurs in even insusceptible neurons. This inducible gene expression system using MAC remains useful for providing critical insights into appearance of TDP-43 pathology.

背景:43 kDa的TAR-DNA结合蛋白(TDP-43)的细胞质错定位是肌萎缩性侧索硬化症(ALS)的主要标志。大多数ALS患者在皮质和脊髓运动神经元中检测到TDP-43聚集;然而,TDP-43错位的病理机制尚不清楚。方法:在小鼠人工染色体(MAC)载体上建立四环素诱导的TDP-43 A315T系统,避免转基因插入突变,建立小鼠胚胎干(ES)细胞系,研究过表达的外源TDP-43 A315T是否错定位于ES细胞源性神经元的细胞质中,并引发对这些细胞的神经毒性作用。结果:可诱导的TDP-43 A315T系统成功加载到MAC上并导入小鼠ES细胞。这些胚胎干细胞可分化为运动神经元和中间神经元。多西环素在两个神经元中过表达TDP-43 A315T,导致细胞质错定位。TDP-43错位导致运动神经元细胞死亡,而中间神经元细胞未发生死亡。结论:TDP-43对细胞质错定位的易感性在神经元类型上是选择性的,而TDP-43过表达的错定位即使在不易感的神经元中也会发生。这种使用MAC的诱导基因表达系统仍然有助于提供对TDP-43病理外观的关键见解。
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