Prevention of core cell damage in isolated islets of Langerhans by low temperature preconditioning

AIM: To study the core cell damage in isolated islets of Langerhans and its prevention by low temperature preconditioning (26 ℃).METHODS: Islets were cultured at 37 ℃ for 7-14 d after isolation, and then at 26 ℃ for 2, 4 and 7 d before additional culture at 37 ℃ for another 7 d. Core cell damage in the isolated islets was monitored by video-microscopy and analyzed quantitatively by use of a computer-assisted image analysis system. The analysis included daily measurement of the diameter and the area of the isolated islets and the area of the core cell damage that developed in those islets over time during culture. Histology and TdT-mediated dUTP-biotin nick end labeling (TUNEL) assay were used to characterize the cell damage and to monitor islet function.RESULTS: Microscopic analysis showed that during the 7 to 14 d of culture at 37 ℃, core cell damage occurred in the larger islets with diameters ＞200 μm, which included both necrotic and apoptotic cell death. Low temperature (26 ℃) culture could prevent core cell damage of isolated islets. The 7-d culture procedure at 26 ℃ could inhibit most of the core cell (excluding diameters＞300 μm) damages when the islets were re-warmed at 37 ℃.CONCLUSION: Our results indicate that core cell damage within isolated islets of Langerhans correlates with the size of islets. Low temperature (26 ℃) culture can prevent core cell damage in isolated islets, and successfully precondition these islets for incubation at 37 ℃. These novel findings may help to understand the pathophysiology of early loss of islet tissue after transplantation, and may provide a new strategy to improve graft function in the clinical setting of islet transplantation.

Prevention of central cell damage to isolated islets of Langerhans in hamsters by low temperature preconditioning

BACKGROUND: The efficacy of clinical islet transplanta- tion has been demonstrated with autografts, and although islet allografts have established insulin independence in a small number of IDDM patients, the treatment is con- founded by the necessity of central cell damage immuno- suppression, the lack of donor tissue, and recurring islet immunogenicity. These limitations underscore a need to develop therapies to serve the large population of diabetic patients. This study was designed to document central cell damage to isolated islets of Langerhans in hamsters and its prevention. METHODS: Islets were cultured at 37 °C for 7-14 days after isolation, and then at 26 °C for 2,4 and 7 days before addi- tional culture at 37 °C for an additional 7 days. Central cell damage in the isolated islets was monitored by video-mi- croscopy and analyzed quantitatively by a computer-assis- ted image analysis system. The analysis included daily measurement of the diameter and the area of the isolated is- lets and the area of the central cell damage that developed in those islets over time during culture. Histological exami- nation and TdT-mediated dUTP-biotin nick end labeling (TUNEL) assay were used to characterize cell damage and to monitor islet function. RESULTS; Microscopic analysis showed that during the 7 to 14 days of culture at 37 °C, central cell damage appeared in the larger islets with diameters greater than 200 μm, which included both necrotic and apoptotic cell death. Low temperature (26 °C) culture prevented central cell damage of isolated islets. The 7-day culture procedure at 26 °C could inhibit most of the central cell ( excluding diameters greater than 300 μm) damage when the islets were re- warmed to 37 °C. CONCLUSIONS: Our results indicate that central cell da- mage to isolated islets of Langerhans correlates with the size of the islets. Low temperature (26 °C) culture can preventcentral cell damage to the isolated islets, and is capable to successfully precondition these islets for 37 °C culture. These novel findings may help to understand the patho- physiology of early loss of islet tissue after transplantation, and may provide a new strategy to improve graft function in the clinical setting of islet transplantation.

Recurrent multisystem Langerhans cell histiocytosis involving the female genitalia: A case report

BACKGROUND Langerhans cell histiocytosis(LCH)is a histiocytic proliferative disease caused by clonal proliferation of Langerhans cells,which is currently defined as an inflam-matory myeloid tumor.It is rare in adults,with an incidence of 1–2 per million,and is highly heterogeneous in clinical presentation,with unpredictable disease progression and outcome.CASE SUMMARY A 52-year-old postmenopausal female patient presented to the gynecology department in July 2023 with bilateral vulvar masses.She was diagnosed with recurrent multisystem LCH.The patient had previously been diagnosed with a single-system and single-focal LCH in October 2021 due to a right maxillofacial mass,which resolved after surgical treatment.A chemotherapy regimen was developed after multidisciplinary consultation.Six cycles of chemotherapy resulted in partial remission,and maintenance chemotherapy is currently being administered.CONCLUSION Recurrent LCH involving the bilateral vulva has been poorly reported.Compre-hensive imaging and pathological evaluation is important for diagnosis.The model of joint multidisciplinary specialist diagnosis and treatment is worthy of clinical application.

Multisystemic recurrent Langerhans cell histiocytosis misdiagnosed with chronic inflammation at the first diagnosis:A case report

BACKGROUND Langerhans cell histiocytosis(LCH)is characterized by diabetes insipidus and is an uncommon occurrence.Pathological biopsies still have a certain degree of diagnostic probability.We present a case in which LCH initially affected the pituitary gland.This resulted in a misdiagnosis of chronic inflammation upon pathological examination.CASE SUMMARY A 25-year-old female exhibited symptoms of diabetes insipidus.Magnetic resonance imaging revealed an enhanced foci in the pituitary gland.After surgical resection of the pituitary lesion,the pathological diagnosis was chronic inflam-mation.However,the patient later experienced bone destruction in the skull and lower limb bones.After the lower limb bone lesion was compared with the initial pituitary lesion,the final diagnosis was modified to LCH.The patient was treated with multiple chemotherapy courses.However,the patient’s condition gradually worsened,and she eventually passed away at home.CONCLUSION LCH should be considered when patients exhibit diabetes insipidus and absence of high signal intensity in the pituitary gland on sagittal T1-weighted image and abnormal enhancement in the pituitary region.

Mannogalactoglucan from mushrooms protects pancreatic islets via restoring UPR and promotes insulin secretion in TIDM mice

Type 1 diabetes mellitus(T1DM) lacks insulin secretion due to autoimmune deficiency of pancreaticβ-cells.Protecting pancreatic islets and enhancing insulin secretion has been therapeutic approaches.Mannogalactoglucan is the main type of polysaccharide from natural mushroom,which has potential medicinal prospects.Nevertheless,the antidiabetic property of mannogalactoglucan in T1DM has not been fully elucidated.In this study,we obtained the neutral fraction of alkali-soluble Armillaria mellea polysaccharide(AAMP-N) with the structure of mannogalactoglucan from the fruiting body of A.mellea and investigated the potential therapeutic value of AAMP-N in T1DM.We demonstrated that AAMP-N lowered blood glucose and improved diabetes symptoms in T1DM mice.AAMP-N activated unfolded protein response(UPR) signaling pathway to maintain ER protein folding homeostasis and promote insulin secretion in vivo.Besides that,AAMP-N promoted insulin synthesis via upregulating the expression of transcription factors,increased Ca^(2+) signals to stimulate intracellular insulin secretory vesicle transport via activating calcium/calmodulin-dependent kinase Ⅱ(CamkⅡ) and cAMP/PKA signals,and enhanced insulin secretory vesicle fusion with the plasma membrane via vesicle-associated membrane protein 2(VAMP2).Collectively,these studies demonstrated that the therapeutic potential of AAMP-N on pancreatic islets function,indicating that mannogalactoglucan could be natural nutraceutical used for the treatment of T1DM.

淋巴结Langerhans细胞组织细胞增生症临床病理及免疫组化研究

目的：研究以淋巴结首发的ＬＣＨ形态结构特征、免疫表型、临床表现及鉴别诊断。方法：应用ＨＥ染色和免疫组化ＡＢＣ法标记Ｏ１０和Ｓ－１００阳性以确定诊断。结果：１４例淋巴结ＬＣＨ有４种组合类型：（１）肾形及卵圆形细胞４例；（２）咖啡豆样细胞３例；（３）咖啡豆样及大圆形细胞５例；（４）大圆形细胞２例。结论：咖啡豆样细胞是ＬＣＨ的特征细胞，具有重要诊断价值，嗜酸性粒细胞是ＬＣＨ重要的组成成分之一。根据细胞形态、分化程度和不同组合，ＬＣＨ分为：①反应性，②交界性，③低度恶性，④高度恶性（肉瘤型）。

高梯度磁场免疫磁珠法分离人表皮Langerhans细胞

采用鼠抗人 OKT6单克隆抗体与包被羊抗鼠 Ig G免疫磁珠的特异性结合 ,利用钢毛产生高梯度磁场 ,进行免疫磁珠法分离纯化人表皮 L C。结果表明 LC纯化率超过 99% ,细胞活性超过 95%。该方法简单、快速、价廉、纯化率高 ,不受样本量的限制 ,易在无菌下分离纯化 。

宫颈癌浸润淋巴细胞和Langerhans细胞免疫组织化学研究

目的：研究宫颈癌浸润淋巴细胞（ＴｉＬ）、Ｌａｎｇｅｒｈａｎｓ细胞（ＬＣ）分布及与淋巴细胞活性标记物ＨＬＡ－ＤＲ和转铁蛋白受体（ＴｆＲ）的关系。方法：应用ＣＤ４，ＣＤ８，ＯＫＴ１１，ＯＫＴ９，ＨＬＡ－ＤＲ和Ｓ－１００抗体及ＡＢＣ免疫组化法。结果：ＣＤ４，ＣＤ８，ＯＫＴ１１阳性细胞随肿瘤发展而增多，并和转铁蛋白受体（ＴｆＲ）及ＨＬＡ－ＤＲ阳性表达密切关联。ＬＣ细胞数目随肿瘤分期增高和ＴｉＬ密度增加而增多。结论：ＴｉＬ和ＬＣ在宫颈癌中数量增多和活性增高的作用尚不清楚，它可能是宫颈癌局部的免疫反应表现。

黄芪增免散对围手术期食管癌组织Langerhans细胞的影响

目的观察黄芪增免散对围手术期食管癌组织Langerhans细胞(LC)的影响。方法采用黄芪增免散治疗围手术期食管癌患者36例,观察手术切除食管组织内LC的变化及形态特征。结果与对照组比较,服药组食管癌组织内LC显著增多(P<0.05),且与癌细胞密切接触,反映对癌细胞杀伤作用增强。随着肿瘤分化程度降低,浸润深度增加,LC及间质淋巴细胞数量明显减少(P<0.01),显示肿瘤对机体免疫功能的损害。结论围手术期食管癌患者免疫调节治疗可改善细胞功能。

食管癌发生过程中Langerhans细胞立体结构的观察

目的 :观察食管癌发生过程中Langerhans细胞 (Langerhanscell,LC)的三维立体结构变化。方法 :采用免疫荧光技术对 2 2例正常食管粘膜、2 0例癌旁组织和 2 3例癌组织中LC进行标记 ,通过激光共聚焦扫描显微镜对LC进行断层扫描和三维立体结构重建。结果 :断层扫描和图象分析显示正常食管粘膜中LC相对面积大 ,形状因子相对值小。癌旁粘膜和癌组织中LC相比面积明显缩小 ,形状因子相对值大 ,正常食管粘膜、癌旁组织和癌组织中LC数量减少 ,胞浆突起逐渐减少 ,立体结构趋向圆形。结论 :食管癌发生过程中LC体积缩小 ,数量减少 ,胞浆突起逐渐减少 ,提示其抗原递呈的功能减退 。

瘢痕疙瘩中Langerhans细胞的电镜观察

目的:观察瘢痕疙瘩组织中Langerhans细胞的超微结构,探讨其与瘢痕疙瘩形成的关系。方法:应用电镜技术观察瘢痕疙瘩及增生性瘢痕切除标本各6例及4例正常皮肤组织中Langerhans细胞的形态,观察其分布特征。结果:电镜下表皮细胞层中朗格罕细胞核形多不规则,胞质中等电子密度,有较多线粒体和溶酶体,扁平囊泡较多。结论:Langerhans细胞在瘢痕疙瘩组织中功能活跃,Langerhans细胞与瘢痕疙瘩的形成关系密切。

Langerhans细胞与瘢痕疙瘩形成的相关性

目的探讨Langerhans细胞与瘢痕疙瘩形成的相关性。方法应用免疫组织化学(ABC法)观察S-100蛋白及CD1a在瘢痕疙瘩及增生性瘢痕与正常皮肤组织中的表达。结果瘢痕疙瘩及增生性瘢痕组织中S-100蛋白及CD1a免疫反应阳性的Langer-hans细胞明显增多。瘢痕疙瘩及增生性瘢痕与正常皮肤之间差异非常显著(P<0.01)。瘢痕疙瘩与增生性瘢痕之间差异不显著(P>0.05)。结论Langerhans细胞与瘢痕疙瘩的形成关系密切。

舌癌中Langerhans细胞的图像分析研究

目的 :探讨Langerhans细胞学与舌癌发生、发展的关系。方法 :采用电子计算机图像分析方法对 30例人类舌癌标本中正常上皮、不典型增生、原位癌及浸润癌ATP酶阳性的Langerhans细胞进行了数量及形态定量研究。结果 :研究发现各期中Langerhans细胞的数量不同 ,不典型增生中最少 (P <0 .0 5 ) ,早期浸润癌数量明显增多 (P <0 .0 5 )。图像分析发现病变各期Langerhans细胞的面密度、体密度、形态因子均较正常上皮中有明显改变 (P <0 .0 5 ) ,但与表面基本相同 (P >0 .0 5 )。结论 :Langerhans细胞数量和形态改变可能与人类舌癌的发生、发展有关。

病理性瘢痕中Langerhans细胞的形态学研究

目的:探讨Langerhans细胞与病理性瘢痕的相关性。方法:应用免疫组织化学法(ABC法)观察瘢痕疙瘩、增生性瘢痕及正常皮肤组织中S-100蛋白和CD1a的表达,用透射电镜观察这三种组织中的Langerhans细胞的超微结构。结果:瘢痕疙瘩及增生性瘢痕组织中对S-100蛋白和CD1a免疫反应阳性的Langerhans细胞较正常皮肤中明显增多。瘢痕疙瘩及增生性瘢痕与正常皮肤之间有显著性差异(P<0.05)。瘢痕疙瘩与增生性瘢痕之间差异不显著(P>0.05)。电镜下表皮细胞层中Langerhans细胞核形多不规则,胞质中等电子密度,有较多线粒体和溶酶体,扁平囊泡较多。结论:Langerhans细胞在瘢痕疙瘩及增生性瘢痕组织中明显增多,且功能活跃。

Langerhans细胞与胆脂瘤型中耳炎

应用ＡＴＰ酶组织化学、免疫组织化学和超微结构等研究手段，观察胆脂瘤表皮铺片及冰冻切片中Ｌａｎｇｅｒｈａｎ细胞（ＬＣ），以及Ｔ＿４、Ｔ＿８、Ｂ细胞和ＨＬＡ－ＤＲ阳性细胞的形态、分布，并通过形态学计量和图象分析得出，胆脂瘤表皮中ＬＣ密度与外耳道皮肤表皮中ＬＣ密度差异无显著性。胆脂瘤基质中Ｔ＿４，平均面数密度和Ｔ＿４／Ｔ＿８比值高于正常人外周Ｔ＿４／Ｔ＿８之比，通过细胞与细胞间相互作用，ＬＣ可能在中耳胆脂瘤发生中起到一定作用。

食管癌Langerhans细胞和AgNOR的研究

目的 研究食管癌Langerhans细胞和AgNOR与病理临床特征的关系,估价其预后意义。 方法 应用S_(-100)蛋白抗体对56例食管癌组织中Langerhans细胞作免疫组化染色。采用胶质银染色技术对癌细胞AgNOR计量观测。 结果 癌组织中Langerhans细胞密度同淋巴细胞反应密度及癌细胞分化程度呈正相关(P<0.05),癌细胞AgNOR计数与其分化有关,分化愈低,计数愈多。Langerhans细胞和AgNOR均与癌大体类型、浸润深度、淋巴结转移状况及患者性别、年龄等特征无关(P>0.05),与患者预后有关,但二者之间相关无显著性(P>0.05。 结论 反映机体免疫状况的Langerhans细胞和反映癌细胞生物学特性的AgNOR对判断癌细胞分化程度和患者预后有意义,值得进一步研究。

食管癌组织中巨噬细胞和Langerhans细胞的免疫组织化学研究

采用ＡＢＣ免疫组化技术，对２４例食管鳞癌组织中的溶菌酶阳性巨噬细胞（Ｌｙｓｏｚｙｍｅｐｏｓｉｔｉｖｅｍａｃｒｏｐｈａｇｅ，ＬＰＭ）和Ｓ－１００蛋白阳性Ｌａｎｇｅｒｈａｎｓ细胞（ＬＣ）进行了原位观察和定量研究。结果显示：在食管粘膜和各级鳞癌组织中均可见到ＩＰＭ及ＬＣ浸润。从食管正常粘膜、异型增生粘膜到癌组织，ＬＰＭ数量逐渐增多（Ｐ＜０．０１或０．０５）。癌组织内ＬＰＭＭ数量与肿瘤的分化程度、大体类型关系密切，提示食管癌组织内浸润的巨噬细胞与食管癌的发生发展有一定的联系。另外，ＬＰＭ与ＬＣ分布明显不同，数量无相关性，说明两者在抗肿瘤免疫中具有不同的功能。

Survival of encapsulated islets: More than a membrane story

At present, proven clinical treatments but no cures are available for diabetes, a global epidemic with a huge economic burden. Transplantation of islets ofLangerhans by their infusion into vascularized organs is an experimental clinical protocol, the first approach to attain cure. However, it is associated with lifelong use of immunosuppressants. To overcome the need for immunosuppression, islets are encapsulated and separated from the host immune system by a permselective membrane. The lead material for this application is alginate which was tested in many animal models and a few clinical trials. This review discusses all aspects related to the function of transplanted encapsulated islets such as the basic requirements from a permselective membrane(e.g., allowable hydrodynamic radii, implications of the thickness of the membrane and relative electrical charge). Another aspect involves adequate oxygen supply, which is essential for survival/performance of transplanted islets, especially when using large retrievable macrocapsules implanted in poorly oxygenated sites like the subcutis. Notably, islets can survive under low oxygen tension and are physiologically active at > 40 Torr. Surprisingly, when densely crowded, islets are fully functional under hyperoxic pressure of up to 500 Torr(> 300% of atmospheric oxygen tension). The review also addresses an additional category of requirements for optimal performance of transplanted islets, named auxiliary technologies. These include control of inflammation, apoptosis, angiogenesis, and the intra-capsular environment. The review highlights that curing diabetes with a functional bio-artificial pancreas requires optimizing all of these aspects, and that significant advances have already been made in many of them.