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This study reports that hairy and enhancer of split homolog-1 (HES1), known to repress gene transcription in progenitor cells of several cell lineages, was strongly expressed in cells and tissues of T-cell lymphoma expressing the oncogenic chimeric tyrosine kinase nucleophosmin (NPM)-anaplastic lymphoma kinase [ALK; ALK

Fusion genes including NPM–ALK can promote T-cell transformation, but the signals required to drive a healthy T cell to become malignant remain undefined. In this study, we introduce NPM–ALK into primary human T cells and demonstrate induction of the epithelial-to-mesenchymal transition (EMT) program, attenuation of most T-cell effector programs, reemergence of an immature epigenomic profile, and dynamic regulation of c-Myc, E2F, and PI3K/mTOR signaling pathways early during transformation. A mutant of NPM–ALK failed to bind several signaling complexes including GRB2/SOS, SHC1, SHC4, and UBASH3B and was unable to transform T cells. Finally, T-cell receptor (TCR)–generated signals were required to achieve T-cell transformation, explaining how healthy individuals can harbor T cells with NPM–ALK translocations. These findings describe the fundamental mechanisms of NPM–ALK-mediated oncogenesis and may serve as a model to better understand factors that regulate tumor formation.Significance:This investigation into malignant transformation of T cells uncovers a requirement for TCR triggering, elucidates integral signaling complexes nucleated by NPM–ALK, and delineates dynamic transcriptional changes as a T cell transforms.See related commentary by Spasevska and Myklebust, p. 3160

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Fusion genes including NPM–ALK can promote T-cell transformation, but the signals required to drive a healthy T cell to become malignant remain undefined. In this study, we introduce NPM–ALK into primary human T cells and demonstrate induction of the epithelial-to-mesenchymal transition (EMT) program, attenuation of most T-cell effector programs, reemergence of an immature epigenomic profile, and dynamic regulation of c-Myc, E2F, and PI3K/mTOR signaling pathways early during transformation. A mutant of NPM–ALK failed to bind several signaling complexes including GRB2/SOS, SHC1, SHC4, and UBASH3B and was unable to transform T cells. Finally, T-cell receptor (TCR)–generated signals were required to achieve T-cell transformation, explaining how healthy individuals can harbor T cells with NPM–ALK translocations. These findings describe the fundamental mechanisms of NPM–ALK-mediated oncogenesis and may serve as a model to better understand factors that regulate tumor formation. Significance: This investigation into malignant transformation of T cells uncovers a requirement for TCR triggering, elucidates integral signaling complexes nucleated by NPM–ALK, and delineates dynamic transcriptional changes as a T cell transforms. See related commentary by Spasevska and Myklebust, p. 3160

ATP citrate lyase (ACLY) is a key metabolic enzyme that catalyzes the generation of Acetyl-CoA and is upregulated in cancer cells and required for their growth. The phosphoinositide 3-kinase (PI3K) and Src-family kinase (SFK) Lyn are constitutively activate in many cancers. We show here, for the first time, that both the substrate and product of PI3K, phosphatidylinositol-(4,5)-bisphosphate (PIP2) and phosphatidylinositol-(3,4,5)-trisphosphate (PIP3), respectively, bind to ACLY in Acute Myeloid Leukemia (AML) patient-derived, but not normal donor-derived cells. We demonstrate the binding of PIP2 to the CoA-binding domain of ACLY and identify the six tyrosine residues of ACLY that are phosphorylated by Lyn. Three of them (Y682, Y252, Y227) can be also phosphorylated by Src and they are located in catalytic, citrate binding and ATP binding domains, respectively. PI3K and Lyn inhibitors reduce the ACLY enzyme activity, the ACLY-mediated Acetyl-CoA synthesis, phospholipid synthesis, histone acetylation and cell growth. Thus, PIP2/PIP3 binding and Src tyrosine kinases-mediated stimulation of ACLY links oncogenic pathways to Acetyl-CoA-dependent pro-growth and survival metabolic pathways in cancer cells.

Background The pneumonia severity index (PSI) scoring system is one of the tools used to evaluate and predict the prognosis of patients with community-acquired pneumonia (CAP). Although PSI has been widely used in clinical studies of pneumonia, it is still rare to combine it with blood indexes to predict the prognosis of pneumonia. Neutrophil-to-lymphocyte ratio (NLR) is a promising candidate predictor of mortality in CAP patients. The aim of this study was to investigate the efficacy of pneumonia severity index combined with NLR in predicting 30-day mortality in CAP patients. Methods We conducted a retrospective study. We analyzed data on 400 non-immune individuals over the age of 18 in this study. All patients received blood routine measurement and PSI score calculation after admission. The primary outcome measures were mortality and survival in CAP patients. The sensitivity and specificity of PSI score, NLR, and the combination of PSI score and NLR in predicting 30-day mortality were assessed using the subject operating characteristic curve (ROC). Results Data from 400 patients were analyzed, in which the 30-day mortality was 10.5% (42/400). The AUC of NLR and PSI in predicting 30-day mortality of CAP patients were 0.81 (95% CI 0.73 - 0.89) and 0.94 (95% CI 0.90 - 0.98), respectively, with statistically significant differences (p = 0.00). The sensitivity and specificity of NLR were 0.80 and 0.7, respectively. The sensitivity and specificity of PSI were 0.78 and 0.94, respectively. The combined AUC of the two indicators for predicting death in CAP patients was 0.95 (95% CI 0.92 - 0.99), and the sensitivity and specificity were 0.85 and 0.94, respectively. Conclusions Neutrophil-to-lymphocyte ratio improves the accuracy and sensitivity of the pneumonia severity index in predicting 30-day mortality of CAP patients.

Background In China, tuberculous pleural effusion is the most common cause for pleural effusion. Elevated ADH and positive tuberculin test usually are characteristic of tuberculous pleural effusion. We reported a 71-year-old male patient with elevated ADH and positive tuberculin test firstly misdiagnosed as tuberculous pleural effusion finally proven as pleural mesothelial sarcoma by thoracoscopic pathology. Methods Appropriate laboratory tests and thoracentesis were carried out. Thoracoscopy and pathological biopsy were performed to differentiate tuberculous pleural effusion. Results Chest CT showed right pleural effusion. ADH in pleural effusion was over 45 U/L and PPD test was positive. No abnormal cells were found in pleural effusion pathology. Pathology of thoracoscopic biopsy proved pleural mesothelioma. Conclusions Elevated ADH and positive tuberculin test are not a specific index for tuberculosis and thoracoscopic biopsy pathology is crucial for differential diagnosis.

Background Detection of carcinoembryonic Antigen (CEA) in pleural effusion has good clinical application value in differentiating benign and malignant pleural effusion, but sometimes CEA provides limited help. We report a case of a patient with left lung neoplasms combined with bilateral pleural effusion with increased CEA in the pleural effusion whose thoracoscopy pleural biopsy pathology was negative, mimicking lung carcinoma and ultimately confirmed as pulmonary sarcomatoid carcinoma by CT-guided percutaneous lung biopsy. Methods The chest computed tomography (CT) scan, thoracoscopy pleural biopsy, and CT-guided percutaneous lung biopsy were arranged to explore the etiology of pleural effusion. Results The chest CT scan showed bilateral pleural effusion with left lung neoplasms, pulmonary atelectasis, and left hilar enlargement. Pathology of thoracoscopy biopsy showed pleural inflammation with infiltration of inflammatory cells. Pathology of CT-guided percutaneous lung biopsy confirmed pulmonary sarcomatoid carcinoma. Conclusions Elevated pleural effusion CEA is not a specific index of lung cancer. CT-guided percutaneous lung biopsy is appropriate for patients presenting with pleural diseases with lung neoplasms, especially when thoracoscopy pleural biopsy result was negative.

BACKGROUND Detection of serum neuron specific enolase (NSE) has high sensitivity and specificity in the diagnosis of lung carcinoma, especially in small cell lung carcinoma, but sometimes serum NSE provided limited help. We report a case of a patient with right lung consolidation combined with elevated serum neuron specific enolase which mimicked lung carcinoma and was ultimately confirmed as pulmonary cryptococcosis by CT-guided percutaneous lung biopsy. METHODS Chest computed tomography (CT) scan and CT-guided percutaneous lung biopsy were performed for diagnosis and blood tests explored the latent etiology. RESULTS The chest CT scan showed right lung consolidation and a pulmonary nodule in lingual segment of upper lobe of left (Figure1A - F). Serum cryptococcal antigen was positive. Pathology of CT-guided percutaneous lung biopsy confirmed pulmonary cryptococcosis (Figure 1G - I). CONCLUSIONS Elevated NSE is not a specific index of lung cancer. Serum cryptococcal antigen and CT-guided percutaneous lung biopsy has high specificity in cryptococcal pneumonia.

While studies have identified a number of mutations in mantle cell lymphoma (MCL), the list may still be incomplete and contribution to the pathogenesis remains unclear. We analyzed the mutational landscape of four mantle cell lymphoma biopsies obtained during an 8-year period from the same patient with his normal cells serving as control; we also established a cell line from the final stage of the disease. Numerous mutations with high allelic burden have been identified in all four biopsies. While a large subset of mutations was seen only in individual biopsies, the core of 21 mutations persisted throughout the disease. This mutational core is also maintained in the cell line that also displays DNA-methylation and cytokine secretion profiles of the primary mantle cell lymphoma cells. This cell line is uniquely sensitive to clinically relevant inhibitors of Bruton's Tyrosine Kinase. The response to Bruton Tyrosine Kinase's inhibition is enhanced by inhibitors of CDK4/6 and mTOR. Among the mutations seen in the primary and cultured MCL cells, mutations of three genes are involved in the control of H3K4 methylation: demethylase KDM5C, present already in the early disease, and methyltransferase KMT2D and cofactor BCOR, both of which are seen late in the disease and are novel and predicted to be pathogenic. The presence of these mutations was associated with hypermethylation of H3K4. Restoration of KDM5C expression affected expression of numerous genes involved in cell proliferation, adherence/movement, and invasiveness.

BACKGROUND Despite the extensive improvement in antibiotic treatment and medical care, severe adult community-acquired pneumonia (CAP) remains as the significant cause of death worldwide. Earlier prognosis assessment and timely treatment in adult CAP patients are useful for prognosis. The neutrophil-to-lymphocyte ratio (NLR) in blood routine has a broad application possibility in assessing inflammatory reaction and prognosis. The aim of this study was to examine the relationship between NLR and inflammatory reaction and to unravel the usefulness of NLR in the assessment of clinical outcomes in adult CAP patients. METHODS This retrospective study was conducted based on adult patients with a primary diagnosis of CAP. All patients included received a routine blood test and calculated NLR. All of the measurement data were analyzed with paired t-test and the enumeration data were analyzed with χ2 test. Multivariate analysis was performed to investigate the association between predictors (age, male, CURB-65 scores, comorbidity, NLR, and other inflammatory cells in blood routine) and unfavorable outcomes of CAP (ICU admission and 30-day mortality). Receiver operating characteristic curves (ROC) were used to evaluate the sensitivity and specificity of NLR in predicting unfavorable outcomes of CAP. RESULTS One hundred fifty patients were included. Compared with favorable outcomes group, age, CURB-65 scores, WBC, neutrophil and lymphocyte counts, and NLR were elevated in unfavorable outcomes group (p < 0.05), gender and coexisting illness did not differ obviously. Multivariate logistic regression model analysis showed CURB-65 scores and NLR were independent predictors correlated with unfavorable outcomes (p < 0.05). The area under the ROC curve (AUC) of NLR was 0.81 (95% CI 0.73 to 0.89), the sensitivity was 81.00% and specificity was 72.8%. NLR is superior to CURB-65 in predicting unfavorable outcomes. NLR combined CURB-65 has better sensitivity and specificity (89.40% versus 91.30%). CONCLUSIONS NLR is a simple, cheap, and rapidly available measurement in blood routine and is associated with unfavorable clinical outcomes in adult CAP patients.

Background High serum neuron-specific enolase (NSE) level has been in use as a tumor marker; however, some physicians may ignore NSE levels in serum, especially when the patients are asymptomatic. Here we report a case that a 51 year old female patient with no respiratory symptoms who had a NSE level which increased extremely over three months and was eventually diagnosed small cell lung cancer (SCLC). Methods Endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) in right pulmonary hilar enlarged lymph node was performed for diagnosis. Results EBUS showed right pulmonary hilar lymph node enlargement. A TBNA biopsy histopathology diagnosed SCLC. Conclusions We should pay attention to high serum NSE levels, especially when the index increased extremely over a short time.

BACKGROUND Community-acquired pneumonia (CAP) is a common infectious disease. Inflammatory reaction and prognosis assessment in adult CAP patients are useful for CAP site of care decisions. Most CAP patients were diagnosed in an out-patient or emergency department, so a simple, cheap and rapidly available measurement to assess inflammatory reaction and prognosis has the prospect of broad application. The aim of this study was to investigate the usefulness of plasma D-Dimer in assessment of inflammatory reaction and prognosis in adult CAP patients. METHODS A retrospective study was conducted. All adult patients with a primary diagnosis of CAP were included and were evaluated by peripheral plasma D-Dimer test. All of the measurement data were analyzed with paired t-test and the enumeration data were analyzed with χ2 test. Correlative factor analysis was performed between D-Dimer levels and serum inflammatory markers (WBC, hs-CRP, PCT) and prognostic indexes (ICU admission and 30-day mortality). Receiver operating characteristic curves (ROC) were used to evaluate the sensitivity and specificity of D-Dimer in predicting ICU admission and/or 30-day mortality. RESULTS One hundred fifty patients were included. Compared with non-D-Dimer elevated group, serum inflammatory markers (WBC, hs-CRP, PCT) and prognostic indexes (ICU admission and 30-day mortality) were elevated in the D-Dimer elevated group (p < 0.05). D-Dimer had positive correlation with serum inflammatory markers (WBC, hs-CRP, PCT), the rates of ICU admission and 30-day mortality, and scores of CURB-65. The AUC of ROC curve of D-Dimer was 0.880 (95% CI 0.823 to 0.936), the sensitivity was 80.4% and specificity was 79.8%, D-Dimer levels are superior to hs-CRP and PCT in predicting 30-day mortality and/or ICU admission according to AUCs of the ROC curves. CONCLUSIONS Elevated plasma D-Dimer in adult CAP patients is associated with an increased inflammatory reaction and ICU admission and 30-day mortality. It can be a simple, cheap, and rapidly available measurement to assess inflammatory reaction and prognosis in adult CAP patients.

pstrongemBackground/em/strong: Scoring systems including CURB-65 and Pneumonia Severity Index (PSI) and novel or traditional biomarkers including procalcitonin (PCT) and c-reactive protein (CRP) are very significant for understanding the severity and prognosis in community-acquired pneumonia (CAP) patients, while prognostic items are useful for CAP prognostication and point-of-care decisions. The aim of this study was to investigate the usefulness of peripheral blood routine items in predicting ICU admission and 30-day mortality in CAP patients./ppstrongemMethods/em/strong: A retrospective study was conducted. All adult patients with a primary diagnosis of CAP were included and peripheral blood routine tests were evaluated. Univariate analysis and multivariate logistic regression analysis were used to explore association of risk factors with 30-day mortality among CAP patients. Receiver operating characteristic curves (ROC) were used to evaluate the sensitivity and specificity of peripheral blood routine items and compared with CURB-65 scores in predicting ICU admission and/or 30-day mortality./ppstrongemResults/em/strong: One hundred fifty patients were included and compared with non-ICU admission patients. There was a statistically significant difference in age, co-existing illness, RDW, WBC, and CURB-65 scores ranking in ICU admission patients (p0.05). In multivariate logistic regression analysis, we found RDW, WBC, and CURB-65 ≥ 3 scores increased the risk of 30-day mortality by 4.01, 1.65, and 3.43 times, respectively. The area under the curve (AUC) of ROC curves of RDW combined with WBC and CURB-65 was 0.786 (95% CI 0.701 to 0.876) and 0.836 (95% CI 0.764 to 0.908), respectively and the sensitivity was 84.0% and 60.0%, respectively, and the specificity 66.7% and 93.7%, respectively./ppstrongemConclusions/em/strong: Elevated RDW and WBC increased mortality in adult CAP patients, RDW combined with WBC had a better sensitivity than CURB-65 scores in predicting ICU admission and/or mortality in CAP patients./p.

Background G-lipopolysaccharide, a component of the cell wall of Gram-negative bacteria, is called lipopolysaccharide. The detection of G-lipopolysaccharide can be used for the early diagnosis of infectious diseases, but some-times G-lipopolysaccharide provides limited help. We report a case of a patient with hemoptysis and high-density shadow of both lungs combined with elevated serum G-lipopolysaccharide which mimicked bronchiectasis with Gram-negative bacterium infection. It was ultimately confirmed as Mycobacterium iranicum infection by CT-guided percutaneous lung biopsy and next generation sequencing. Methods The chest computed tomography (CT) scan, CT-guided percutaneous lung biopsy, and NGS were performed for diagnosis and blood tests explored for the latent etiology. Results The chest CT scan showed a high-density shadow of both lungs, atelectasis of right middle lobe, multiple enlarged lymph nodes in mediastinum and right hilum. Pathology of CT-guided percutaneous lung biopsy indicated fibrous tissue proliferation and granulation tissue formation and some alveolar epithelial cells slightly proliferated with focal carbon powder deposition in alveolar sacs and spaces. The lung tissue NGS confirmed Mycobacterium iranicum infection. Conclusions Elevated serum G-lipopolysaccharide is not a specific index for infectious diseases. CT-guided percutaneous lung biopsy and lung tissue NGS has high specificity in pathogen detection of infectious diseases.

Background: To report an atypical case misdiagnosed as lung abscess over the past 2 months, but persistent anemia combined with significantly increased hs-CRP and lung lesions indicated systemic lesion, which led to the diagnosis of granulomatosis with polyangiitis proven by lung biopsy and anti-neutrophil cytoplasmic antibody test (ANCA). Methods: The complete blood count, hs-CRP, and anti-neutrophil cytoplasmic antibody (ANCA) test were performed. The pathology consultation for the lung biopsy was arranged. Results: Hemoglobin was 8.5 g/L, hs-CRP was > 200 mg/L, c-ANCA directed against anti-proteinase 3 (PR3) was positive, pathology consultation reported granulomatous inflammation. Conclusions: When patients have multiple organ dysfunction combined with anemia and significantly increased hs-CRP, physicians should pay attention to systemic vasculitis. .

Background To report a case of broncholithiasis with recurrent hemoptysis and fever initially misdiagnosed as active tuberculosis. Methods The chest contrast-enhanced CT scan, electronic bronchoscope, and ultrathin bronchoscope were performed leading to the diagnosis of broncholithiasis, open lung lobectomy was done after thoracic surgery consultation. Results The chest contrast-enhanced CT scan showed a high-density intratracheal shadow and calcified lymph nodes. Ultrathin bronchoscopy manifested calcified lesions located at the distal portion of the right lower lobe bronchus. Histopathology of lobectomy showed lithiasis in the right lower lobe tracheobronchial tree. Conclusions We should pay attention to calcified intratracheal lesions and make differential diagnosis with tuberculosis, especially when accompanied with calcified lymph nodes and fever.

iBackground:/iTracheobronchopathia osteochondroplastica (TO) is a rare benign disease. We report a case of TO.iMethods:/iChest CT scan and bronchoscope with biopsy was performed for diagnosis and blood tests explored for the latent etiology.iResults:/iChest CT scan and bronchoscopic images showed multiple nodular protrusions in the trachea and main bronchi. Histopathology demonstrated sub-mucosal ossification and inflammatory cell infiltration. Laboratory inspection showed adenovirus and coxsackie B virus IgM antibodies were positive.iConclusions:/iThe patient recently had a virus infection and inflammation was observed in histopathology, which indicated adenovirus and coxsackie B virus may play a role in the occurrence or exacerbation of TO.

iBackground:/iCryptococcal pneumonia is an uncommon lesion in immune-competent adults. Histological evidence of Cryptococcus neoformans is a gold criterion for diagnosis. Here we report a case firstly misdiagnosed as tuberculosis from a lung biopsy.iMethods:/iChest computed tomography (CT) scan and CT-guided puncture were performed for diagnosis and blood tests explored for the latent etiology.iResults:/iChest CT scan images showed multiple nodules in the left peripheral lower lobe. Histopathology demonstrated multiple granulomatous inflammatory response lacking evidence of Cryptococcus neoformans, acid-fast staining was negative, serum cryptococcal antigen was positive.iConclusions:/iSerum cryptococcal antigen has high specificity in cryptococcal pneumonia.

A fundamental feature of tumor progression is reprogramming of metabolic pathways. ATP citrate lyase (ACLY) is a key metabolic enzyme that catalyzes the generation of Acetyl-CoA and is upregulated in cancer cells and required for their growth. The phosphoinositide 3-kinase (PI3K) and Src-family kinase (SFK) Lyn are constitutively activate in many cancers. We show here, for the first time, that both the substrate and product of PI3K, phosphatidylinositol-(4,5)-bisphosphate (PIP2) and phosphatidylinositol-(3,4,5)-trisphosphate (PIP3), respectively, bind to ACLY in Acute Myeloid Leukemia (AML) patient-derived, but not normal donor-derived cells. We demonstrate the binding of PIP2 to the CoA-binding domain of ACLY and identify the six tyrosine residues of ACLY that are phosphorylated by Lyn. Three of them (Y682, Y252, Y227) can be also phosphorylated by Src and they are located in catalytic, citrate binding and ATP binding domains, respectively. PI3K and Lyn inhibitors reduce the ACLY enzyme activity, ACLY-mediated Acetyl-CoA synthesis, phospholipid synthesis, histone acetylation and cell growth. Thus, PIP2/PIP3 binding and Src tyrosine kinases-mediated stimulation of ACLY links oncogenic pathways to Acetyl-CoA-dependent pro-growth and survival metabolic pathways in cancer cells. These results indicate a novel function for Lyn, as a regulator of Acetyl-CoA-mediated metabolic pathways.

Mutations in leucine-rich repeat kinase 2 (LRRK2) are common causes of familial Parkinson's disease (PD). LRRK2 has been shown to bind peroxiredoxin-3 (PRDX3), the most important scavenger of hydrogen peroxide in the mitochondria, in vitro. Here, we examined the interactions of LRRK2 and PRDX3 in Drosophila models by crossing transgenic LRRK2 and PRDX3 flies. As proof of principle experiments, we subsequently challenged LRRK2 and LRRK2/PRDX3 flies with a peroxidase mimic, Ebselen. We demonstrated that co-expression of PRDX3 with the LRRK2 kinase mutant G2019S in bigenic Drosophila ameliorated the G2019S mutant-induced reduction in peroxidase capacity, loss of dopaminergic neurons, shortened lifespan and mitochondrial defects of flight muscles in monogenic flies expressing the G2019S alone. Challenges with Ebselen recapitulated similar rescue of these phenotypic features in mutant-expressing Drosophila. The peroxidase mimic preserved neuronal and mitochondrial and neuronal integrity and improved mobility and survival in mutant-expressing Drosophila. Taken together, our study provides the first in vivo evidence to suggest that phosphoinhibition of endogenous peroxidases could be a mechanism in LRRK2-induced oxidant-mediated neurotoxicity. Our therapeutic experiments also highlight the potential of thiol peroxidases as neuroprotective agents in PD patients carrying LRRK2 mutations.

We examined the response of the temperature coefficient (Q10) for soil respiration to changes in soil temperature and soil moisture through a laboratory incubation experiment. Two types of soils differing in vegetation and moisture status were collected and incubated under two temperatures (10 and 30 8C) and two soil moisture regimes (35 and 75% of water holding capacity, WHC) for 5 weeks. Before and after the incubation experiment, the temperature coefficient of soil respiration was measured using sodalime method by changing temperature in a water bath. For both soils, the mean Q10 values of the respiration rate were 2.0 in the 30 8C and 2.3 in the 10 8C soil treatments. Higher temperature with lower soil moisture treatment significantly decreased the Q10 value, whereas lower temperature with higher soil moisture treatment significantly enhanced the Q10 value (ANOVA, p 0.05). Although incubation temperature and moisture accounted for 40 and 29% (as r 2 � 100%), respectively, of variations in Q10, soil water-soluble carbon content alone could have explained 79% of the variation, indicating that the availability of respiratory substrate, rather than the pool of soil microorganisms, played a crucial role in the response of the temperature coefficient to environmental factors. These results suggest that biotic factors should also be taken into consideration when using the Q10 function to predict the response of soil respiration to global warming. # 2005 Elsevier B.V. All rights reserved.

The endometrial-myometrial interface (EMI) is a specific functional region of uterus. However, our knowledge on EMI ultrastructure both in normal uterus and adenomyosis is far from enough to understand its pathology. In this study, used the samples of EMI and outer myometrium (OM) from the adenomyosis hysterectomy specimens and the subjects from the control uteri, we prospectively compared the ultrastructure of myocytes from EMI and OM, the ultrastructural changes of EMI between the proliferative and secretory phases, and the ultrastructural difference of EMI between adenomyosis and the control group. In both adenomyosis and control group, there were differences in ultrastructure between myocytes from EMI and OM. Specifically, the myocytes from EMI were rich in organelles. In contrast, the myocytes from OM had abundant contractile structural components. In the proliferative phase, the myocytes from EMI in adenomyosis had significantly smaller cell and nucleus diameter than those from the control group, but in the secretory phase, the difference was not significant. In the control group, the various ultrastructural features of myocytes from EMI including the mean diameter of cell and nuclei and the myofilaments/cytoplasm ratio exhibited cyclical changes, but in adenomyosis, the normal cyclical changes were absent. In conclusions, there are significant ultrastructural differences between the myocytes from EMI and OM. The myocytes in women with adenomyosis were significantly different to the control subjects, primarily because the normal cyclical changes were absent.

In this paper, we consider a highly recursive interconnection network known as the fully connected cubic network (FCCN). By exploiting its recursive properties, we thoroughly analyze the performance of a simple routing algorithm for the FCCN. We show that at least 80% of the routes obtained from this simple algorithm are shortest paths, and this percentage increases further with the network size. Subsequently, we obtain the network diameter and average internodal distance, taking into account the communication locality that is exhibited in many parallel computations. The presence of the communication locality significantly reduces the average internodal distance.

Background/Aims: Apoptosis is a common mechanism for the regulation of cell loss. It is associated with both tissue atrophy and metaplasia. Cigarette smoking has tremendous adverse effects on the stomach and also increases the risk of gastric cancer. This action may be through the change in apoptosis in the stomach. The aim of this study was to examine the effect of cigarette smoking on apoptosis in the gastric mucosa and the possible role of epidermal growth factor (EGF) in this action. Methods: Gastric blood flow was assessed by the laser Doppler technique. Serum and gastric mucosal EGF levels were measured by RIA. Gastric mucosal apoptosis was determined using TdT-mediated dUTP-biotin nick end labeling (TUNEL). Results: Cigarette smoke exposure decreased serum EGF which was accompanied by a reduction in gastric blood flow. Meanwhile, gastric mucosal cell apoptosis was increased. Administration of EGF (20 μg/kg i.v.) before each cigarette smoke exposure reversed these actions. Removal of salivary glands induced similar effects on the gastric blood flow, apoptosis, and serum EGF level as with cigarette smoke exposure. Conclusion: A reduction in serum EGF was involved in the decrease in gastric blood flow and increase in gastric mucosal apoptosis caused by cigarette smoking.

Here we report that T-cell lymphomas characterized by the expression of anaplastic lymphoma kinase (ALK+ TCL) fail to express the TNFalpha and frequently display DNA methylation of the TNFalpha gene promoter. While only a subset of the ALK+ TCL-derived cell lines showed a high degree of the promoter methylation, all 6 showed low to nondetectable expression of the TNFalpha mRNA, and none expressed the TNFalpha protein. All 14 ALK+ TCL tissue samples examined displayed some degree of the TNFalpha promoter methylation, which was the most prominent in the distal portion of the the promoter. Treatment with a DNA methyltransferase inhibitor, 5'-aza-2'-deoxy-cytidine (5-ADC), reversed the promoter methylation and led to the expression of TNFalpha mRNA and protein. Furthermore, in vitro DNA methylation of the promoter impaired its transcriptional activity in the luciferase reporter assay. This impairment was seen even if only either distal or proximal portion were methylated, with methylation of the former exerting a more profound inhibitory effect. Notably, the ALK+ TCL cell lines uniformly expressed the type 1 TNFalpha receptor (TNF-R1) protein known to transduce the TNFalpha-induced pro-apoptotic signals. Moreover, exogenous TNFalpha inhibited growth of the ALK+ TCL cell lines in a dose-dependent manner and induced activation of the members of the cell apoptotic pathway: Caspase 8 and caspase 3. These findings provide additional rationale for the therapeutic inhibition of DNA methyltransferases in ALK+ TCL. They also suggest that treatment with TNFalpha may be highly effective in this type of lymphoma.

Although STAT5A and STAT5B have some nonredundant functional properties, their distinct contributions to carcinogenesis are not clearly defined. Here we report that STAT5A expression is selectively inhibited by DNA methylation of the STAT5A gene promoter region in cells expressing the oncogenic tyrosine kinase NPM1-ALK (also known as NPM-ALK). The DNA methylation is induced by NPM1-ALK itself via STAT3, and is associated with binding to the promoter of the gene encoding MeCP2 capping protein and with lack of binding of the STAT5A gene transcription activator SP1. Reversal of methylation by the DNA methyltransferase inhibitor 5'-aza-2'-deoxycytidine restores SP1 binding and STAT5A gene expression. Notably, the induced or exogenously expressed STAT5A protein binds to the enhancer and intron 14 of the NPM1-ALK gene and triggers selective suppression of NPM1-ALK expression. These results show that NPM1-ALK induces epigenetic silencing of STAT5A gene and that STAT5A protein can act as a key tumor suppressor by reciprocally inhibiting expression of NPM1-ALK.

In this study, we demonstrated that STAT3, a well-characterized transcription factor expressed in continuously activated oncogenic form in the large spectrum of cancer types, induces in malignant T lymphocytes the expression of DNMT1, the key effector of epigenetic gene silencing. STAT3 binds in vitro to 2 STAT3 SIE/GAS-binding sites identified in promoter 1 and enhancer 1 of the DNMT1 gene. STAT3 also binds to the promoter 1 region and induces its activity in vivo. Treatment of the malignant T lymphocytes with STAT3 siRNA abrogates expression of DNMT1, inhibits cell growth, and induces programmed cell death. In turn, inhibition of DNMT1 by a small molecule inhibitor, 5-aza-2-deoxy-cytidine, and 2 DNMT1 antisense DNA oligonucleotides inhibits the phosphorylation of STAT3. These data indicate that STAT3 may in part transform cells by fostering epigenetic silencing of tumor-suppressor genes. They also indicate that by inducing DNMT1, STAT3 facilitates its own persistent activation in malignant T cells. Finally, these data provide further rationale for therapeutically targeting STAT3 in T-cell lymphomas and, possibly, other malignancies.

Expression of SHP-1 phosphatase, a key negative regulator of cell signaling, is lost in T cell lymphomas and other malignancies due to DNA methylation of the SHP-1 promoter by a currently undefined mechanism. We demonstrate that malignant T cells express DNA methyltransferase (DNMT) 1 and that constantly activated signal transducer and activator of transcription (STAT) 3 is capable of bindingin vitroto DNA oligonucleotides corresponding to four STAT3 SIE/GAS binding sites identified in the SHP-1 promoter. STAT3, DNMT1, and histone deacetylase 1 form complexes and bind to the SHP-1 promoterin vivo. Treatment with pharmacologic grade DNMT1 anti-sense oligonucleotides and STAT3 small-interfering RNA induces in the malignant T cells DNA demethylation and expression ofSHP-1gene. These data indicate that STAT3 may, in part, transform cells by inducing epigenetic silencing of SHP-1 in cooperation with DNMT1 and, apparently, histone deacetylase 1. Reversal of such gene silencing represents an attractive aim for novel anticancer therapies.