NeuroD1 and Ascl1 Convert Human Glial Cells into Neurons in Ex Vivo Culture of Human Brain Tissue
Liang XuQingsong WangJiancheng LiaoJiajun ZhengBing QinWen LiJiaxuan ZhangWei LiXiangyu WangMaoying ZhangGong Chen
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Abstract Transcription factor-mediated cell conversion has been reported in the central nervous system (CNS) of both rodents and non-human primates (NHPs). In particular, glia-to-neuron (GtN) conversion has been achieved in the brains and spinal cords of animal models of neurological disorders for neural regeneration and repair. However, whether GtN conversion can ultimately be used for human brain repair in patients is still unknown. To investigate the applicability of GtN conversion technology in the human brain, we established a long-term ex vivo culture system using human brain tissue that was surgically removed from epileptic patients to directly test GtN conversion. We demonstrate that both neural transcription factors NeuroD1 and Ascl1 can convert human glial cells into neurons. Importantly, both immunostaining and electrophysiological recordings revealed that the glia-converted neurons showed immature properties during the initial 1–2 weeks of conversion, and then acquired more mature neuronal properties after 3–4 weeks of conversion. These ex vivo conversion studies in human brain tissue pave the way toward future clinical trials using a transcription factor-based glia-to-neuron conversion approach to treat neurological disorders.Keywords:
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Human brain
日本消化器外科学会雑誌Vol. 45 (2012) No. 7 p. 724-731 大野 吏輝ほか著「胆道気腫を契機に診断された十二指腸原発T/NK cell lymphoma の1 例」におきまして，誤りがございましたので，お詫びとともに下記のとおり訂正いたします． p. 724 《誤》 現病歴：2010 年8 月上旬より腹痛が出現し近医を受診した． 《正》 現病歴：2009 年8 月上旬より腹痛が出現し近医を受診した． p. 728 Fig. 6 図説 《誤》 Histological findings: histochemical staining (×20). A: Immunostaining for CD20 was negative. B: Immunostaining for CD3 was positive. C: Immunostaining for TIA-1 was positive. 《正》 Histological findings: histochemical staining (×20). A: Immunostaining for CD3 was positive. B: Immunostaining for CD20 was negative. C: Immunostaining for TIA-1 was positive. p. 730 《誤》 ……1 手術例を経過した．本疾患は比較的新しい概念の範疇であり，…… 《正》 ……1 手術例を経験した．本疾患は比較的新しい概念の範疇であり，……
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Мета. Покращення результатів лікування хворих з місцево-розповсюдженими та рецидивними солідними пухлинами черевної порожнини та заочеревинного простору.
Матеріали і методи. За період з червня 2015 по січень 2018 р. в Національному інституті раку виконали комбіновані оперативні втручання з нефректомією 28 хворим з приводу первинних місцево-розповсюджених та рецидивних солідних пухлин черевної порожнини та заочеревинного простору.
Результати. У 5 із 28 пацієнтів виконали нефректомію ex vivo ex situ з аутотрансплантацією нирки, у 4 - успішно. Гостре ушкодження нирок спостерігали у 6 (26%) хворих, яким аутотрансплантації нирки не виконували. Після операції померли 2 (8,7%) хворих. У пацієнтів, яким нирка була збережена, не спостерігали гострого ушкодження нирок, ніхто з цих пацієнтів не помер.
Висновки. З метою профілактики розвитку гострого ушкодження та хронічної хвороби нирок у майбутньому можливість виконання аутотрансплантації нирки у разі хірургічного лікування солідних пухлин черевної порожнини та заочеревинного простору, окрім первинного раку нирки, повинна бути розглянута щодо кожного хворого. Дану процедуру доцільно виконувати в спеціалізованих лікувальних закладах, де накопичено досвід в онковаскулярній хірургії.
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Antigen Recovery for Vimentin Immunostaining by Reducing Agents for Two Cases of Spindle Cell Tumors
AbstractThree clinical cases in which imlnunostaining on formalin fixed, paraffin embedded tissue resulted in falsenegative or only rare staining for vilnentin were pretreated with various methods. For 2 of these cases, treatment with reducing agents improved the extent of immunostaining for vimentin and added only 23 min to the staining procedure. Pretreatment with microwave irradiation has been previously reported to improve immunostaining for vinientin. However for 1 of the 2 cases in which reducing agents enhanced vimentin immunostaining, pretreatment with microwave irradiation was ineffective at improving vimentin staining. This result indicates that it may be helpful to use a reducing agent in parallel with the microwave in-adiation technique when the results of immunostaining for vimentin are important for the differential diagnosis. (The J Histotechnol 21:45, 1998)Keywords: antigen recoveryantigen retrievalenhanced immunostainfalse-negativeimmunohistochemistryimmunostain enhancementvimentin
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The latent membrane protein (LMP) of Epstein-Barr virus can be detected by an immunostaining method in about 50% of Hodgkin's disease lymph node tumour cells on frozen sections. The same method had been successfully employed on paraffin-embedded (PE) sections. We used this immunostaining method on imprints (IP) and PE sections of 26 Hodgkin's lymph nodes. Overall, there were 11 LMP-positive cases; immunostaining was stronger in IP than in PE samples; in nine cases both IP and PE samples were LMP-positive; in two, imprints were LMP-positive when PE sections were LMP-negative.
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Background: A proliferating trichilemmal cyst (PTC) is an uncommon, rapidly-reproducing cutaneous epithelial cyst, differentiating from the isthmic portion of the outer hair root sheath. It is usually described as a benign tumor, but malignant transformation has been reported and is then denominated as a malignant proliferating trichilemmal tumor. Ki67 immunostaining has been used as a methodology for the evaluation of tumor grade in other tumors, due to its distinctive reaction patterns which exclusively involve proliferating cells. Objectives: (1) To report the incidence of cases of PTCs in a General Hospital during a 12 years period. (2) To determine the expression of ki67 using immunohistochemical staining. (3) To correlate ki67 reaction patterns with clinical prognosis. Materials and Methods: The dermatology department's files during a period of 12 years were reviewed; cases with a diagnosis of PTC were selected, and ki67 immunostaining was done when enough biological material was available. Results: A total of 15 cases with a diagnosis of PTC were identified. In 12 cases, ki67 immunostaining was carried out. In 9 of the 12 cases, ki67 was observed in the basal cells of the cystic epithelium, one case was moderately positive in palisading epithelial cells; in the other two cases ki67 immunostaining was negative. Clinical follow-up was done and revealed that no patient had local recurrence in 5 years after surgical removal of PTC. We therefore consider this immunostaining technique is probably correlated with low recurrence potential.
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Mammary gland tumors (MGTs) are common in dogs and show a variable clinical behavior that is difficult to predict. Currently, few immunohistochemical markers have been established to predict the prognosis of a canine MGT. However, p53 immunostaining has been variably reported to be prognostic for canine MGTs. Additionally, while p16CDK2NA protein (p16) immunostaining has been found to be prognostic for human breast cancers, this marker has never been evaluated as a prognostic marker for canine neoplasms. In the present study, the prognostic utility of p53 and p16 was evaluated in 35 canine malignant MGTs. It was observed that 19 (54%) dogs died due to their MGTs with an overall mean survival time (MST) of 882 days. Seven MGTs showed p53 immunostaining, but this was not significantly associated with death (4 of 7 vs. 15 of 28; p = 0.6) or MST (670 vs. 934 days; p = 0.57). Five dogs had MGTs with no p16 immunostaining, 28 MGTs had intermediate p16 immunostaining, and two MGTs had increased p16 immunostaining. Neither death due to MGT (4 of 5, 14 of 28, or 1 of 2; p = 0.28) nor MST (683, 927, and 307 days; p = 0.31) were significantly associated with p16 immunostaining. Interestingly, p53 immunostaining was significantly associated with an increase or loss of p16 immunostaining. This is the first time that p16 has been evaluated as a prognostic marker for canine neoplasms. While these results suggest that a proportion of canine MGTs develop by cellular mechanisms that alter both p53 and p16 expression, there was no evidence that defects in p53 or p16 alter the behavior of a MGT. Neither p53 nor p16 was found to significantly predict prognosis, although this could reflect the limited number of MGTs included in the study.
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p16INK4a immunohistochemistry (IHC) is widely used to facilitate the diagnosis of human papillomavirus (HPV)-associated neoplasia, when ≥70% of cells show strong nuclear and cytoplasmic positivity. In this study, we aim to compare partial expression patterns that do not fulfill the above criteria and seek biological implications in laryngeal squamous cell carcinoma (LSCC).p16INK4a IHC staining was conducted on representative sections of archived tissue from 88 LSCCs. Immunoreactivity was described based on four parameters: intracellular localization of immunostaining, intensity of immunostaining, distribution pattern and percentage of positive cells.Six patterns of p16INK4a immunoexpression were observed and defined as: strong diffuse (strong immunostaining, expression in cytoplasm and nucleus in >70% of tumor cells), weak diffuse (moderate or weak immunostaining, expression in cytoplasm in >70% of tumor cells), marginal (strong cytoplasmic immunostaining, limited to the periphery of tumor islets), strong scattered (strong immunostaining, expression in cytoplasm and nucleus in <50% of tumor cells), weak scattered (moderate or weak immunostaining, expression in cytoplasm in <50% of tumor cells), negative (no expression). The pN stage of the patients was associated with p16INK4a immunoexpression patterns, the marginal pattern was only found in the pN0-Nx stages, while the weak diffuse pattern was more frequently observed in pN2-N3 stages.Partial immunostaining with architecturally distinct p16INK4a immunoexpression patterns may prove significant in stratifying characteristic clinicopathological subgroups among LSCC. Our observations may support the hypothesis that p16INK4a has different roles in different subcellular locations, with tumorigenic molecular pathways unrelated to HPV infection.
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Immunostaining refers to the processes of localizing antigens in cells, thus exploiting the principle that antibodies bind specifically to antigens in biological tissues. There are two general methods of immunostaining: immunohistochemistry (IHC) and immunofluorescense. For human embryonic stem cells (hESCs), researchers use immunostaining to detect specific undifferentiation markers, to demonstrate pluripotency by embryoid body (EB) and by teratoma formation (immunostaining with three germ layer markers), and to detect specific differentiation processes.
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Although intraoperative rapid diagnosis is conventionally performed using hematoxylin–eosin (HE)-stained specimens, the use of additional special staining, together with immunostaining techniques, has been examined in recent years to improve diagnostic accuracy. In intraoperative rapid diagnosis, immunostaining should be completed within 7–10 min, because the pathologist is typically presented with an HE-stained specimen within the same time period. We hypothesized that ultrasound may enhance antigen–antibody reactions and reduce the number of immunostaining steps. To clarify the ability of ultrasound to support immunostaining, we first created an ultrasonic generator specifically for immunostaining. Next, we explored the optimal conditions for immunostaining of formalin-fixed specimens to examine the utility of the ultrasonic generator. Finally, we tried immunostaining with the ultrasonic generator using frozen specimens to simulate intraoperative rapid diagnosis. We report herein that ultrasound enables immunostaining of frozen specimens in ×10 min.
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