Complex structures and diverse functions of nuclear bodies: a review / 生物工程学报

Nuclear bodies are membrane-free nuclear substructures that are localized in the mammalian nuclear matrix region. They are multiprotein complexes that recruit other proteins to participate in various cellular activities, such as transcription, RNA splicing, epigenetic regulation, tumorigenesis and antiviral defense. It is of great significance to clarify the functions and regulatory mechanisms of nuclear bodies to probe related diseases and virus-host interactions. This review takes several nuclear bodies associated proteins as examples, summarizes the formation process, structure and functions of nuclear bodies, and focuses on their important roles in antiviral infection. It is expected to provide new insight into host antiviral mechanisms.

Post-translational Modifications of PML in Regulating the Functions of Nuclear Bodies --Review / 中国实验血液学杂志

Abstract　　The promyelocytic leukemia (PML) gene encoded PML protein as a tumor suppressor protein, plays important roles in the occurrence and development of various cancers including acute promyelocytic leukemia. Recent studies have indicated that there are a variety of post-translational modifications of the PML protein, such as SUMOylation, ubiquitination, phosphorylation, and acetylation in cells. These modifications of the PML protein can directly affect the formation of PML nuclear bodies (PML-NBs), repair DNA damage, and modulate cell apoptosis. Furthermore, the abnormal modifications of PML not only result in the occurrence of hematopoietic tumors, but also are closely related to the drug-resistance of cancer. Therefore, investigating the post-translational modifications of PML is significant to uncover the mechanism of formation and functions of PML-NBs, thus contributing to the prevention and treatment of related hematopoietic tumors. In this review, the characteristics of the post-translational modifications of PML protein and the relationship between these modifications and functions of PML-NBs are summarized so as to provide the potential targets for the treatment of related cancers.

Morphologic Analysis of Cytomegalovirus Infected Cells in Bronchial Washing Cytology: Comparison of Liquid-Based Preparation and Conventional Smear

BACKGROUND: The cytopathic effects of cytomegalovirus (CMV) infection have been well described since the virus was first reported; however, the morphology of CMV infection has not been clearly studied. We examined the difference in detailed cytologic findings in bronchial washing cytology between liquid-based and conventionally prepared smears. METHODS: Bronchial washing cytology was processed using either the conventional preparation (CP) or liquid-based preparation (LBP). Sixty-nine cells with typical cytopathic effects of CMV infection were detected on CP slides and 18 cells on LBP slides. Using the image analyzer, area, circumference, major axis, and minor axis of the cytoplasm, nucleus, and intranuclear inclusion were measured in singly scattered CMV-infected cells, and histiocytes were used as a control. RESULTS: The mean cytoplasmic area of CMV-infected cells was 1.47 times larger than that of histiocytes in CP and 2.92 times larger in LBP (p<.05). The mean nuclear area of CMV-infected cells was 2.61 times larger than that of histiocytes in CP and 4.25 times larger in LBP (p<.05). The nucleus to cytoplasm ratio and intranuclear inclusion to cytoplasm ratio of the mean area, circumference, major axis, and minor axis in CP were larger than those in LBP (p<.05). CONCLUSIONS: The sizes of cytoplasm, nucleus, and intranuclear inclusion were larger in LBP than in CP, indicating that CMV-infected cells are easily detectable in LBP. However, the nucleus-to-cytoplasm ratio was larger in CP, suggesting that differentiation from malignancy or regenerative atypia requires caution in CP.

Analysis of Histologic Features Suspecting Anaplastic Lymphoma Kinase (ALK)-Expressing Pulmonary Adenocarcinoma

BACKGROUND: Since 2007 when anaplastic lymphoma kinase (ALK) rearrangements were discovered in non-small cell lung cancer, the ALK gene has received attention due to ALK-targeted therapy, and a notable treatment advantage has been observed in patients harboring the EML4/ALK translocation. However, using ALK-fluorescence in situ hybridization (FISH) as the standard method has demerits such as high cost, a time-consuming process, dependency on interpretation skill, and tissue preparation. We analyzed the histologic findings which could complement the limitation of ALK-FISH test for pulmonary adenocarcinoma. METHODS: Two hundred five cases of ALK-positive and 101 of ALK-negative pulmonary adenocarcinoma from January 2007 to May 2013 were enrolled in this study. The histologic findings and ALK immunohistochemistry results were reviewed and compared with the results of ALK-FISH and EGFR/KRAS mutation status. RESULTS: Acinar, cribriform, and solid growth patterns, extracellular and intracellular mucin production, and presence of signet-ring-cell element, and psammoma body were significantly more often present in ALK-positive cancer. In addition, the presence of goblet cell-like cells and presence of nuclear inclusion and groove resembling papillary thyroid carcinoma were common in the ALK-positive group. CONCLUSIONS: The above histologic parameters can be helpful in predicting ALK rearranged pulmonary adenocarcinoma, leading to rapid FISH analysis and timely treatment.

Cytological Features of a Lymphoepithelial Cyst Collected from Fine Needle Aspiration of the Thyroid Gland That Mimicked Papillary Thyroid Carcinoma: A Case Report

Since its first description in 1989, lymphoepithelial cyst of the thyroid gland (LEC-T) has been generally considered a branchial cleft derivative similar to its presentation in other sites, including thymus, parathyroid, and pancreas. However, its characterization has mainly focused on histologic and sonographic findings, and cytological findings are generally described simply or left out entirely. Fine needle aspiration cytology (FNAC) of our case showed large tissue fragments of epithelial cells on a background of lymphocytes. Some areas showed sheets or small nests of squamoid cells, which were closely admixed with clusters of lymphoid cells. Squamous cells contained relatively moderate to large amounts of eosinophilic cytoplasm and vesicular nuclei with occasional nuclear grooves that lacked intranuclear inclusions. Some cells demonstrated keratinization and nuclear atypia. Herein, we describe FNAC findings of LEC-T and review other possible diagnoses.

Successful Treatment of Pneumonia caused by Aspergillus terrerus and Cytomegalovirus after Chemotherapy for Acute Myelogenous Leukemia

We report a case of pneumonia caused by Aspergillus terreus and cytomegalovirus (CMV) in a patient with acute myleogenous leukemia (AML) after remission induction chemotherapy. A 19-year-old woman underwent chemotherapy for AML. Twenty-three days after completing chemotherapy, she experienced a neutropenic fever with a rapidly-progressive pulmonary infiltration. In those days, her serum galactomannan immunoassay was 4.7 and she was treated with intravenous voriconazole (6 mg/kg q12h for 2 doses, followed by 4 mg/kg q12h) because of persistent fever and radiological worsening, despite the administration of amphotericin B deoxycholate (1 mg/kg q24h) for 7 days. A chest CT showed wedge-shaped consolidation with a central hypodense lesion and an air-crescent sign in the right middle lobe. With maintenance therapy of oral voriconazole for 10 weeks, a partial response was shown and neutrophil count was still less than 100/mm3. A lobectomy of the right middle lobe was performed. A. terreus was discovered from the lung tissue. At the same time, giant cells with intranuclear inclusions were found and immunohistochemical staining for CMV was positive. Ganciclovir (5 mg/kg q12h) was added to voriconazole therapy for 3 weeks after surgery, and then cord blood hematopoietic stem cell transplantation (HSCT) was performed. During HSCT, foscarnet (60 mg/kg q12h) was substituted for ganciclovir, and both antiviral agents were used alternatively due to CMV DNAemia. After 83 days from HSCT, the patient achieved successful engraftment and discharged without worsening the pneumonia.

Cytologic Features of Giant Cell Ependymoma: A Case Report and Review of the Literature

Here, we present a case of anaplastic giant cell ependymoma (GCE) occurring in a 15-year-old woman. Squash smear slides for intraoperative frozen section diagnosis revealed oval to round cell clusters with a papillary structure in a fibrillary background. This was occasionally accompanied by the presence of bizarre pleomorphic giant cells with hyperchromatic nuclei and prominent intranuclear inclusions. These intranuclear inclusions were a key clue to diagnosis of ependymoma. Histologic analysis revealed features of a high-grade tumor with perivascular pseudorosettes and bizarre pleomorphic giant cells, which established the diagnosis of GCE. We performed a review of literatures about the cytologic features of GCE, including our case, thus proposing that intraoperative frozen diagnosis of GCE would be established by squash smear preparations featuring the mitosis and necrosis, as well as the high cellularity, and the presence of giant cells showing hyperchromatic nuclei with eosinophilic cytoplasm and intranuclear inclusions/pseudoinclusions.

Fine Needle Aspiration Cytology of Metastatic Adenocarcinoma of the Gingiva from the Lung: A Case Report

Metastases of malignant tumors to the oral region from distant sites are uncommon. A 45-year-old man with painless gingival swelling was diagnosed with adenocarcinoma of the lung. On cytology, clusters of tumor cells on mucous background revealed enlarged nuclei, indistinct cell borders, and irregular nuclear membranes. Some cells showed nuclear inclusions, nuclear grooves and small nucleoli. These findings are indicative of metastatic adenocarcinoma. We present a case of gingival metastasis from a lung adenocarcinoma.

The potential link between PML NBs and ICP0 in regulating lytic and latent infection of HSV-1

Herpes simplex virus type 1 (HSV-1) is a common human pathogen causing cold sores and even more serious diseases. It can establish a latent stage in sensory ganglia after primary epithelial infections, and reactivate in response to stress or sunlight. Previous studies have demonstrated that viral immediate-early protein ICP0 plays a key role in regulating the balance between lytic and latent infection. Recently, It has been determined that promyelocytic leukemia (PML) nuclear bodies (NBs), small nuclear sub-structures, contribute to the repression of HSV-1 infection in the absence of functional ICP0. In this review, we discuss the fundamentals of the interaction between ICP0 and PML NBs, suggesting a potential link between PML NBs and ICP0 in regulating lytic and latent infection of HSV-1.

E2FBP1 antagonizes the p16(INK4A)-Rb tumor suppressor machinery for growth suppression and cellular senescence by regulating promyelocytic leukemia protein stability / 国际口腔科学杂志·英文版

Cellular senescence is an irreversible cell cycle arrest triggered by the activation of oncogenes or mitogenic signaling as well as the enforced expression of tumor suppressors such as p53, p16(INK4A) and promyelocytic leukemia protein (PML) in normal cells. E2F-binding protein 1 (E2FBP1), a transcription regulator for E2F, induces PML reduction and suppresses the formation of PML-nuclear bodies, whereas the down-regulation of E2FBP1 provokes the PML-dependent premature senescence in human normal fibroblasts. Here we report that the depletion of E2FBP1 induces the accumulation of PML through the Ras-dependent activation of MAP kinase signaling. The cellular levels of p16(INK4A) and p53 are elevated during premature senescence induced by depletion of E2FBP1, and the depletion of p16(INK4A), but not p53 rescued senescent cells from growth arrest. Therefore, the premature senescence induced by E2FBP1 depletion is achieved through the p16(INK4A)-Rb pathway. Similar to human normal fibroblasts, the growth inhibition induced by E2FBP1 depletion is also observed in human tumor cells with intact p16(INK4A) and Rb. These results suggest that E2FBP1 functions as a critical antagonist to the p16(INK4A)-Rb tumor suppressor machinery by regulating PML stability.

A Case of Duodenal Ulcer Due to Coinfection with Strongyloides stericoralis and Cytomegalovirus

Gastrointestinal strongyloidiasis and Cytomegalovirus infection mostly occur in patients receiving cancer chemotherapy, undergoing immunosuppressive therapy after organ transplantation, and suffering from acquired immune deficiency syndrome. A 56-year-old man was admitted to the hospital because of abdominal pain and constipation. He had a 10-year history of chronic obstructive pulmonary disease and has been treated intermittently with systemic steroid. The gastroduodenoscopic examination showed a single ulcer on the duodenal bulb and microscopic finding of the biopsy specimens from the ulcer revealed Strongyloides stercoralis and cytomegalovirus immunohistochemical stain positive intranuclear inclusion body on the mucosal surface. The patient was successfully treated with albendazole and ganciclovir.

A case of cytomegalovirus colitis in a patient with liver cirrhosis / 대한내과학회지

Most cases of cytomegalovirus (CMV) colitis are associated with an immunocompromised status, especially with human immunodeficiency virus (HIV) infections and transplantation. It rarely occurs in immunocompetent adults. We report a rare case of CMV colitis in a patient with HBV related liver cirrhosis. A 49 year-old patient presented with lower abdominal pain and bloody diarrhea. Sigmoidoscopic examination showed multiple shallow ulcerations and severe mucosal edema. Conservative treatment for the patient including antibiotics was not effective. Repeated sigmoidoscopic examination with re-biopsy showed intranuclear inclusion bodies suggestive of CMV infection. After administration of Ganciclovir, abdominal pain and bloody diarrhea improved.

The Cytologic Features of Desmoplastic Small Round Cell Tumor with Intranuclear Inclusions : A Case Report

Desmoplastic small round cell tumor (DSRCT) is a rare neoplasm of young adults and it is characterized by polyphenotypic differentiation. We experienced a case of abdominal DSRCT that occurred in a 19-year-old female who presented with painful swelling of her right forearm. The tumor was cytokeratin-negative and it exhibited some tumor cells with intranuclear inclusions. Molecular demonstration of EWS-WT1 fusion transcripts is particularly useful to confirm the diagnosis of DSRCT without epithelial differentiation. We report here on a case of cytokeratin-negative DSRCT that showed an unusual feature of intranuclear inclusions.

Spontaneous Pneumothorax as an Initial Manifestation of Metastatic Papillary Thyroid Carcinoma

Spontaneous pneumothorax occurs in a variety of lung diseases. Primary or metastatic lung cancers are rare, but important causes of pneumothorax. We report here on a case of pulmonary metastasis of thyroid papillary carcinoma that initially presented as spontaneous pneumothorax. A 32-year-old male with no history of thyroid disease underwent wedge resection of the lung due to recurrent pneumothorax. Histologically, the pleura and subpleural lung parenchyma revealed several micronodular lesions and randomly scattered glandular structures with mild cytological atypia. The cells were characterized by their overlapped hypochromatic nuclei with longitudinal grooves and inconspicuous nucleoli. On the additional sections, we found a few intranuclear inclusions and colloid-like material in the follicular structures. The glandular cells were immunoreactive for thyroglobulin, TTF-1, CK 7 and HMW-CK and they were focally immunoreactive for EMA, but they were negative for S100, chromogranin and CEA. The Ki-67 labelling index was 4%. Neck ultrasonography revealed an ill-defined hypoechoic nodule in the left lobe of the thyroid. The diagnosis of conventional, classical papillary carcinoma was made by both the cytology and the total thyroidectomy specimen. Pathologists should conduct a through histologic examination for the patients with recurrent spontaneous pneumothorax to search for the underlying causes.

Cytomegalovirus infection with lissencephaly.

Lissencephaly is a malformation of the brain in which the brain surface is smooth, rather than convoluted. Among the various causes of lissencephaly, infection by a virus during pregnancy plays an important role. Cytomegalovirus (CMV) is an important pathogen causing this anomaly. We present this case of a young female with 24-week-gestation diagnosed on ultrasound as carrying an anomalous fetus with lissencephalic features. At autopsy, there were multiple intra-nuclear CMV inclusions in the brain and the kidneys. This case is presented for its rarity and for the documentation of the tissue localization of CMV inclusions at autopsy.

Fine Needle Aspiration Cytology of Langerhans Cell Histiocytosis in a Lymph Node: A Case Report / 대한세포병리학회지

Langerhans cell histiocytosis (LCH) is a pleomorphic disease entity characterized by local or disseminated atypical Langerhans cells (LCs) found most commonly in bone, lungs, mucocutaneous structures and endocrine organs. Among different sites, unifocal disease confined to a lymph node (LN) is rare. We report a case of LCH confined to a LN in a 38-year-old male who presented with a 2 cm-sized cervical mass. The fine needle aspiration (FNA) smears of cervical LN showed high cellularity having isolated LCs with contorted nuclei and nuclear grooves mixed in multinucleated giant cells, small lymphocytes and eosinophils. Charcot-Leyden crystals were also seen, as were a few dendritic-like cells and intranuclear inclusions. Confirmation of LCH was made by histopathologic studies, positive reactions for S-100 protein and CD1a immunohistochemical staining and by the demonstration of Birbeck granules on electron microscopy. The differentials to be considered include dermatopathic lymphadenitis, sinus histiocytosis with massive lymphadenopathy, Hodgkin's lymphoma and malignant histiocytosis. The characteristic cytomorphologic pattern of LCH in a LN FNA smear plays an important role in suggesting the diagnosis of LCH.

Parvovirus B19 infection-associated hemophagocytic lymphohistiocytosis / 대한내과학회지

Hemophagocytic lymphohistiocytosis is an unusual syndrome that's characterized by fever, hepatosplenomegaly, cytopenias, hypertriglyceridemia, hypofibrinogenemia and pathologic finding of hemophagocytosis in the bone marrow and other tissues. A previously healthy 16-year-old male was admitted because of fever. The cervical and axillary lymph nodes, liver and spleen were palpable. CBC revealed pancytopenia with a decreased reticulocyte count, and the bone marrow aspiration smear showed the presence of giant pronormoblasts with intranuclear inclusion bodies and an increased number of histiocytes that were engulfing blood cell. IgM antibody against Parvovirus B19 and PCR for Parvovirus B19 were positive. Thus, he was diagnosed with hemophagocytic lymphohistiocytosis that was caused by Parvovirus B19 infection. Parvovirus B19 is an agent rarely associated with hemophagocytic lymphohistiocytosis, and in most cases it occurs in those patients with an underlying disease. We report here on a case of hemophagocytic lymphohistiocytosis associated with acute Parvovirus B19 infection in healthy male.

Pure Red Cell Aplasia (PRCA) due to Parvovirus B19 Infection after Kidney Transplantation / 대한이식학회지

The patient was a 39-year-old male with anemia persistent after a living-related renal transplantation. He was diagnosed to have pure red cell aplasia (PRCA) due to parvovirus B19 infection on the 6th week after the renal transplantation. Serum Parvovirus B19 DNA polymerase chain reaction (PCR) and anti-Parvovirus B19 IgM were positive and bone marrow aspiration biopsy showed giant pronormoblasts including prominent intranuclear inclusions. He has been receiving immunosuppressive therapy including oral cyclosporine A, prednisolone, mycophenolate mofetil (MMF). After diagnosis of pure red cell aplasia, we reduced the dose of cyclosporine A and maintained prednisolone, mycophenolate mofetil. We used intravenous immunoglobin(IVIG) 0.4 g/kg/ day for 5 days. Patient's serum reticulocyte count increased a week after the treatment from 0.1% to 3.8%, and patient's serum hemoglobin level normalized on the 4th week of the treatment. Presently, 20 weeks following the initiation of IVIG, his hemoglobin remains normal without recurrent symptom. We are planning to follow up the serum anti-parvovirus B19 IgM/IgG and parvovirus B19 DNA PCR examination.

Multiple Glomus Tumors of the Ankle with Prominent Intranuclear Pseudoinclusions

Glomus tumors are neoplasms that are composed of modified smooth muscle cells of the glomus body. Here, we report a case of multiple glomus tumors of the ankle that showed various histologic types, including the solid type (glomus tumor proper) and angiomatous type (glomangioma). The tumor cells observed in this case also showed prominent intranuclear inclusions, which has not yet been reported in glomus tumors. Ultrastructural examination demonstrated that the nuclear inclusions were not true inclusion bodies but were intranuclear cytoplasmic pseudoinclusions formed by cytoplasmic invaginations that formed as a result of the deep and complex nuclear contours.

Therapeutic Strategies in Huntington's Disease

This article provides an overview of the therapeutic strategies, from ordinary classical drugs to the modern molecular strategy at experimental level, for Huntington's disease. The disease is characterized by choreic movements, psychiatric disorders, striatal atrophy with selective small neuronal loss, and autosomal dominant inheritance. The genetic abnormality is CAG expansion in huntingtin gene. Mutant huntingtin with abnormally long glutamine stretch aggregates and forms intranuclear inclusions. In this review, I summarize the results of previous trials from the following aspects; 1. symptomatic/palliative therapies including drugs, stereotaxic surgery and repetitive transcranial magnetic stimulation, 2. anti-degenerative therapies including anti-excitotoxicity, reversal of mitochondrial dysfunction and anti-apoptosis, 3. restorative/reparative therapies including neural trophic factors and tissue or stem cell transplantation, and 4. molecular targets in specific and radical therapies including inhibition of truncation of huntingtin, inhibition of aggregate formation, normalization of transcriptional dysregulation, enhancement of autophagic clearance of mutant huntingtin, and specific inhibition of huntingtin expression by sRNAi. Although the strategies mentioned in the latter two categories are mostly at laboratory level at present, we are pleased that one can discuss such "therapeutic strategies", a matter absolutely impossible before the causal gene of Huntington's disease was identified more than 10 years ago. It is also true, however, that some of the "therapeutic strategies" mentioned here would be found difficult to implement and abandoned in the future.