Long-term survival outcomes of patients with primary ocular adnexal MALT lymphoma: A large single-center cohort study.

BACKGROUND: Primary ocular adnexal extranodal marginal zone mucosa-associated lymphoid tissue lymphoma (OAML) is a rare subtype of non-Hodgkin's lymphoma, and no consensus has been defined concerning the optimal treatment strategies. This study aims to investigate the associations of disease characteristics and different treatments with long-term outcomes of patients with localized OAML. METHODS: A large retrospective cohort study was conducted in a single-center of China, and 166 patients with newly diagnosed primary localized OAML were enrolled. Detailed data of disease characteristics at diagnosis and treatments were collected for all patients. We compared treatment response and progression-free survival (PFS) among patients with different characteristics and treatments. RESULTS: Of the 166 patients, 52 received complete resection of neoplasm, whereas 114 had residual lesion after surgery. Among the 114 patients, 61 underwent watchful waiting and 53 received further treatment including localized radiotherapy, chemotherapy, or combined radiotherapy and chemotherapy. Median follow-up was 49 months. A total of 31 patients had disease progression or relapse, including four patients with such event more than five years after initial treatment. The 5-year PFS was 73.9%, 70.6%, and 85.9%, whereas the 10-year PFS was 69.3%, 59.2%, and 79.3%, among patients with complete resection of neoplasm, patients in the watchful waiting group and patients with further treatment, respectively. Patients with further treatment had longer PFS, compared with patients in the watchful waiting group (p = 0.011). Bilateral involvement at diagnosis was associated with significantly inferior PFS (p = 0.029), whereas age, IPI score, or TNM staging were not associated with PFS. No serious adverse reaction was reported among patients with further treatment. CONCLUSIONS: Bilateral involvement was associated with poor prognosis. Among patients with residual lesions after surgery, further treatment was associated with improved survival. Patients with OAML might experience disease progression or relapse more than five years after initial treatment.

LncRNA BCYRN1-induced autophagy enhances asparaginase resistance in extranodal NK/T-cell lymphoma.

Background: Asparaginase (ASP) is the cornerstone drug in the treatment of extranodal NK/T-cell lymphoma (ENKTCL), and the mechanisms of resistance to ASP remain largely unknown. Long non-coding RNAs play important roles in chemotherapy resistance in various cancers. However, the expression of BCYRN1 and its role in ENKTCL still remain unidentified. Methods: Lentivirus-mediated BCYRN1 overexpression and knockdown were performed in SNK-6 cells. Cell autophagy was analyzed by adenovirus expressing GFP-LC3B fusion protein. RNA pull-down and RNA Binding Protein Immunoprecipitation Assay were performed to investigate the relationship between BCYRN1 and p53. Western blot analysis was performed to assess the effect of BCYRN1 on different autophagy pathways. Finally, in vivo xenograft tumor model was constructed to analyze the effect of BCYRN1 on tumor growth and ASP resistance. Results: BCYRN1 was overexpressed in ENKTCL than normal NK cells, and patients with higher expression had significantly inferior progression-free survival (PFS). The IC50 value of ASP was significantly increased in BCYRN1-overexpressed SNK-6 cells and BCYRN1 overexpression could resist the inhibitory effect of ASP on proliferation. ASP could induce concurrent apoptosis and autophagy in ENKTCL, and the latter process was enhanced by overexpression of BCYRN1, mainly through affecting both PI3K/AKT/mTOR and p53/mTOR pathways. BCYRN1 could induce the degradation of p53 via ubiquitination, thus resulting in enhancement of autophagy and ASP resistance, which could be reversed by drug-induced autophagy inhibition. The effect of BCYRN1 on tumor growth and autophagy were confirmed in vivo xenograft model. Conclusions: It was found that BCYRN1 was a valuable prognostic biomarker in ENKTCL. BCYRN1 could promote resistance to ASP by inducing autophagy, which could be reversed by inhibition of autophagy. Our findings highlight the feasibility of combining autophagy inhibition and ASP in the treatment of ENKTCL.

Cordycepin suppresses cell proliferation and migration by targeting CLEC2 in human gastric cancer cells via Akt signaling pathway.

PURPOSE: Gastric cancer is a common malignancy worldwide, and is associated with high morbidity and mortality rates. Cordycepin is a 3'-deoxyadenosine drug with significant anti-cancer effects. The aim of this study was to determine the molecular mechanisms underlying cordycepin action on gastric cancer cell proliferation and migration. METHODS: The human gastric cancer cell lines MGC-803 and HGC-27 were treated with different concentrations of cordycepin (25 µM, 50 µM, 100 µM and 5 µM, 25 µM and 50 µM) for 48 h. Cell proliferation was assessed by MTT and colony formation assays, and in vitro migration by the wound healing and transwell assays. In addition, Flow Cytometry was used to detect the cell cycle and apoptosis. RT-PCR and Western blotting were used to evaluate the expression levels of key factors. RESULTS: Cordycepin significantly inhibited gastric cancer cell proliferation and migration in a dose-dependent manner, in addition to inducing apoptosis and arresting the cell cycle at the G2 phase. Mechanistically, cordycepin targeted the PI3K/Akt signaling pathway by significantly altering the expression levels/activation of several key mediators, and upregulated the anti-metastatic factor CLEC2. CONCLUSION: Cordycepin inhibited the proliferation and migration of gastric cancer cells by upregulating CLEC2 via the Akt signaling pathway.

Effect of OSW-1 on microRNA expression profiles of hepatoma cells and functions of novel microRNAs.

3ß,16ß,17α-trihydroxycholest-5-en-22-one 16-O-(2-O-4-methoxybenzoyl-ß-D-xylopyranosyl)-(1→3)- (2-O-acet​yl-α-L-arabinopyranoside) (OSW-1) is a member of the cholestane saponin family, which was first isolated from the bulbs of Ornithogalum saundersiae and previously reported to be cytotoxic against several types of malignant cells. However, its antitumor mechanism remains unclear. Therefore, we investigated microRNA (miRNA) expression profiles in order to explore the antitumor activities of OSW-1. Furthermore, following study of differentially expressed miRNAs, the function of novel miRNAs and OSW-1 was determined using known miRNAs and anticarcinogens. The present study demonstrated that treatment with OSW-1 leads to the upregulation and downregulation of a large set of tumor-related miRNAs, including miR-299, miR-1908, miR-125b, miR-187a, miR-1275, hav1-miR-H6-3p, miR-181, miR-210, miR-483, miR-126, miR-208 and others. Notably, miR-141, miR-142, miR-200C and miR-1275 were found to be upregulated by OSW-1 and doxorubicine, as compared with doxorubicine alone. Additionally, the expression fold-change of miR-142-3P was ~58 times higher than its expression with a different treatment. These miRNAs are linked to cancer, including proliferation, differentiation, apoptosis, cell adhesion, migration, polarity and epithelial to mesenchymal transition (EMT).

[Clinicopathologic study of invasive fungal rhinosinusitis caused by Aspergillus and Mucorales].

OBJECTIVE: To compare the differences in clinicopathologic features of invasive fungal rhinosinusitis caused by Aspergillus and Mucorales, and to discuss the pathogenesis of tissue injury induced by these two kinds of fungi. METHODS: The clinical and pathologic features of 19 patients with invasive fungal rhinosinusitis due to Aspergillus (group A) and 16 patients with invasive fungal rhinosinusitis due to Mucorales (group M) were retrospectively reviewed. HE, PAS and GMS stains were performed on all the paraffin-embedded tissues. The diagnosis was confirmed by histologic examination and microbiological culture results. RESULTS: Amongst the group A patients, the clinical course was acute in 4 cases and chronic in 15 cases. Thirteen cases had underlying predisposing conditions, including diabetes (number = 4), malignant tumor (number = 5), history of trauma (number = 1) and radical maxillary sinus surgery (number = 3). Follow-up information was available in 13 patients. Seven of them died, 4 due to fungal encephalopathy and 3 due to underlying diseases. Amongst the group M patients, the clinical course was acute in 14 cases and chronic in 2 cases. Fourteen cases had underlying predisposing conditions, including diabetes (number = 8), malignant tumor (number = 5) and history of wisdom tooth extraction (number = 1). Follow-up information was available in 14 patients. Four of them died of fungal encephalopathy. There was significant difference in clinical onset between the two groups (P = 0.01). There was however no difference in terms of underlying predisposing conditions and disease mortality. Histologically, the microorganisms in group A patients formed fungal masses and attached to the mucosal surface, resulting in necrotic bands (11/19). Epithelioid granulomas were conspicuous but multinucleated giant cells were relatively rare. Deep-seated necrosis, granulomatous inflammation against fungal organisms (3/19) and vasculitis with thrombosis (4/19) were not common. On the other hand, large areas of geographic necrosis involving deep-seated tissue could be seen in group M patients (13/16). Isolated multinucleated giant cells were commonly seen. Granulomatous inflammation against fungal organisms were identified (16/16). Vasculitis and thrombosis were also observed (10/16). CONCLUSIONS: The invasiveness of Mucorales is remarkable; and when it causes invasive fungal rhinosinusitis, the clinical course is often acute and large areas of tissue necrosis can be seen. The invasiveness of Aspergillus in tissue is relatively mild. Granulomas are more common and the disease often runs a chronic clinical course. There is however no significant difference in long-term mortality. The pathogenesis may be related to the different components of the fungi.

[Methylation of p16 and hMLH1 genes in esophageal squamous cell carcinoma and reflux esophagitis].

OBJECTIVE: To study the promoter methylation pattern of p16 and hMLH1 genes in esophageal squamous cell carcinoma and reflux esophagitis, and to correlate the results with clinical and pathologic findings. METHODS: Twelve cases of normal esophagus, 13 cases of esophageal squamous cell carcinoma, 43 cases of reflux esophagitis with basal cell hyperplasia and 21 cases of reflux esophagitis with dysplasia, as confirmed by endoscopic and pathologic examination, were enrolled into the study. Genomic DNA was extracted. The promoter methylation status of p16 was measured by methylation-specific polymerase chain reaction. The promoter methylation status of hMLH1 was measured by sodium bisulfite-restriction enzyme digestion. Immunohistochemical study for p16 and hMLH1 proteins was also carried out. RESULTS: The rates of p16 methylation in normal esophageal epithelium, basal cell hyperplasia, dysplasia and esophageal squamous cell carcinoma were 0/12, 14.0% (6/43), 38.1% (8/21) and 6/13, respectively. The p16 methylation correlated with the progress of esophageal lesions. On the other hand, the hMLH1 methylation was not observed in the normal esophageal epithelium and reflux esophagitis. One case of esophageal squamous cell carcinoma showed the presence of hMLH1 methylation. The hMLH1 promoter hypermethylation did not correlate with the clinical and pathologic features. CONCLUSIONS: The p16 methylation may be one of the earliest events in the pathogenesis of esophageal squamous cell carcinoma and is also observed in reflux esophagitis. Reflux esophagitis may be related to the development of esophageal squamous cell carcinoma in Chinese population. In contrast, hMLH1 methylation may not be directly involved in the tumorigenesis of esophageal squamous cell carcinoma.

[Clinicopathologic study of malignant tumors in head and neck region complicated by fungal infection].

OBJECTIVE: To study the clinicopathologic features of malignant tumors in head and neck region complicated by fungal infection. METHODS: Twenty-one cases of malignant tumors occurring in head and neck region complicated by fungal infection were retrieved from the archival file. The light microscopic findings were reviewed. Histochemical (for PAS and GMS) and immunohistochemical (for MUC5B) studies were carried out. Fungal culture results were available in 13 of the 21 cases. RESULTS: The age of the patients ranged from 12 to 72 years (median = 48 years). The male-to-female ratio was 17:4. Eight cases (38.1%) were complicated by invasive fungal sinusitis, with orbital involvement in 6 cases and brain involvement in 1 case. The primary tumors in such cases included leukemia (n = 7) and nasopharyngeal carcinoma (n = 1). The fungi belonged to Zygomycete in 5 cases and Aspergillus in 3 cases. These patients had history of chemotherapy/radiotherapy or antibiotics usage. The remaining 13 cases of fungal infection often affected necrotic tumor tissue in nasal cavity, paranasal sinuses, pharynx, larynx and palate. The fungi involved were Aspergillus (n = 6) and Candida (n = 4). Seven of such patients had received radiotherapy. Fungal culture was positive in 9 cases. Fourteen patients had follow-up information available and six of them died of the disease. CONCLUSIONS: Malignant tumors occurring in head and neck region can be complicated by fungal infection. Invasive fungal sinusitis (due to Zygomycetes and Aspergillus) often occurs in patients with leukemia, tends to involve orbit and is associated with poor prognosis. On the other hand, Aspergillus and Candida are the commonest fungi found in the necrotic tumor tissue. Pathologic examination remains the hallmark in confirming the diagnosis and fungal typing.

[Clinicopathologic evaluation of 36 cases of allergic fungal sinusitis].

OBJECTIVE: To characterize clinicopathological features of allergic fungal sinusitis (AFS). METHODS: Thirty-six cases of AFS were retrieved from the department archival files of Beijing Tongren Hospital from 2002 to 2006. AB-PAS, GMS and MUC5B stain were performed using paraffin-embedded tissues of the cases. Ten cases with available fresh diagnostic tissue were investigated by electron microscopy. RESULTS: Patients included 21 males and 15 females. The age of patients ranged from 11 to 53 years. Atopy was very common in these patients. On plain CT scans, the affected nasal sinuses were filled with soft tissue shadow with patchy hyperdensity. The bony sinus wall showed areas of pressure erosion. Skin antigen tests showed fungal positivity in 31 of 36 cases. Serum levels of the total IgE and/or the specific fungal IgE were elevated in 20 cases. The eosinophil quantity was elevated in 23 cases. Fungal culture was positive in 10 cases. Gross examination showed thick putty secretions within the lesions. Light microscopy showed typical "eosinophilic mucin". Fungal elements were seen with AB-PAS, GMS and MUC5B stains. Electron microscopy demonstrated degranulation by the eosinophils. CONCLUSIONS: "Eosinophilic mucin" is the typical histopathological feature of AFS. AB-PAS, GMS and MUC5B staining methods can used to detect fungal species in mucin. Accurate diagnosis of AFS requires correlations among clinical findings, radiologic examinations, laboratory tests and histopathologic features. However, the ultimate diagnosis requires a histopathologic confirmation.

Induction and down-regulation of Sox17 and its possible roles during the course of gastrointestinal tumorigenesis.

BACKGROUND & AIMS: The activation of Wnt/beta-catenin signaling causes the development of gastric and colon cancers. Sox17 represses Wnt/beta-catenin signaling and is down-regulated in colon cancer. This study was designed to elucidate the role of Sox17 during the course of gastrointestinal tumorigenesis. METHODS: Sox17 expression was examined in gastrointestinal tumors of mouse models and humans. The roles of Sox17 in gastric tumorigenesis were examined by cell culture experiments and by construction of Sox17 transgenic mice. RESULTS: Sox17 was induced in K19-Wnt1/C2mE mouse gastric tumors and K19-Wnt1 preneoplastic lesions, where Wnt/beta-catenin signaling was activated. Consistently, Wnt activation induced Sox17 expression in gastric cancer cells. In contrast, Sox17 was rarely detected by immunohistochemistry in gastric and colon cancers, whereas strong nuclear staining of Sox17 was found in >70% of benign gastric and intestinal tumors. Treatment with a demethylating agent induced Sox17 expression in gastric cancer cells, thus indicating the down-regulation of Sox17 by methylation. Moreover, transfection of Sox17 in gastric cancer cells suppressed both the Wnt activity and colony formation efficiency. Finally, transgenic expression of Sox17 suppressed dysplastic tumor development in K19-Wnt1/C2mE mouse stomach. CONCLUSIONS: Sox17 plays a tumor suppressor role through suppression of Wnt signaling. However, Sox17 is induced by Wnt activation in the early stage of gastrointestinal tumorigenesis, and Sox17 is down-regulated by methylation during malignant progression. It is therefore conceivable that Sox17 protects benign tumors from malignant progression at an early stage of tumorigenesis, and down-regulation of Sox17 contributes to malignant progression through promotion of Wnt activity.

[Significance of MUC5B antibody in differential diagnosis between Aspergillus species and Mucorales of fungal sinusitis].

OBJECTIVE: To differentiate between Aspergillus species and Mucorales of fungal sinusitis by immunohistochemistry. METHODS: Formalin-fixed paraffin-embedded tissue blocks of 66 cases of fungal sinusitis were retrieved from the archival files of Department of Pathology of Beijing Tongren Hospital during the period from 2001 to 2006. The samples included 29 cases of fungal balls, 12 cases of allergic fungal sinusitis, 24 cases of chronic invasive fungal sinusitis and 1 case of acute invasive fungal sinusitis. The types of fungi were 44 Aspergillus species (31 cases of A. fumigatus, 7 cases of A. flavus and 6 cases of A. terreus) and 22 Mucorales (14 cases of Mucor species and 8 cases of Rhizopus species). Immunohistochemistry was performed with MUC2, MUC5AC and MUC5B antibodies. The results were compared with histochemical study for periodic acid-Schiff (PAS) and Grocott methenamine silver (GMS) stains. RESULTS: Immunohistochemical study for MUC5B showed that the positive rate of Aspergillus species was 90.9%, in contrast to 4.5% in Mucorales (P < 0.001). The expression of MUC2 and MUC5AC was completely negative, whereas PAS and GMS stains were positive in all cases. CONCLUSION: MUC5B antibody appears to be a useful immunohistochemical marker for identifying fungal types in tissue sections, especially in distinguishing between Aspergillus species and Mucorales in fungal sinusitis.

The use of MUC5B antibody in identifying the fungal type of fungal sinusitis.

Fungal sinusitis is an opportunistic fungal infection. Candida albicans, Aspergillus spp, and Mucorales, the most common pathogenic fungi, differ in both prognosis and therapy. Early diagnosis and differentiation between the subtypes therefore are pivotal for adequate treatment. This report describes a diagnostic immunohistochemical method that is able to distinguish these types of fungi. Formalin-fixed paraffin-embedded blocks of 89 fungal sinusitis specimens (12 C albicans, 52 Aspergillus spp, and 25 Mucorales) and 21 cultures (5 C albicans, 11 Aspergillus spp, and 5 Mucorales) were stained with MUC2, MUC5AC, and MUC5B antibodies. The immunohistochemical staining results of paraffin-embedded samples for MUC5B were successful in 100% and 92.3% of the C albicans and Aspergillus spp samples, respectively. Only 4.0% of the Mucorales parafin sections were found positive, demonstrating a significant difference in detection from C albicans and Aspergillus spp (P < .001). MUC5B expressions for cultures showed that it stained 100% and 90.9% for C albicans and Aspergillus spp, respectively, but negative for Mucorales. The expressions of MUC2 and MUC5AC for both paraffin-embedded samples and cultures were negative. The present study demonstrates the ability of an MUC5B antibody to distinguish C albicans and Aspergillus spp from Mucorales and its use as a diagnostic tool in fungal sinusitis.

Platelet-type 12-lipoxygenase accelerates tumor promotion of mouse epidermal cells through enhancement of cloning efficiency.

Accumulating evidence suggests that platelet-type 12-lipoxygenase (p12-LOX) plays an important role in tumor development. However, how p12-LOX contributes to tumorigenesis is still not understood. The role of p12-LOX was therefore examined in tumor promotion using mouse epidermal JB6 P+ cells that are sensitive to 12-O-tetradecanoylphorbol-13-acetate-induced transformation. The expression of p12-LOX was significantly higher in JB6 P+ cells than in JB6 P- cells that were resistant to transformation, and its expression was further increased by tumor necrosis factor (TNF)-alpha. Importantly, the inhibition of p12-LOX in JB6 P+ cells by baicalein, a specific inhibitor or small interfering RNA significantly suppressed TPA-induced transformation. Moreover, treatment with 12(S)-hydroxyeicosatetraenoic acid (HETE), a metabolite of p12-LOX, enhanced TPA-induced neoplastic transformation either in the presence or absence of baicalein. These results indicate that p12-LOX is required for tumor promotion of epidermal cells and that 12(S)-HETE functions as a rate-limiting factor. Notably, treatment with baicalein significantly suppressed the proliferation of JB6 P+ cells when cells were seeded at a low density in a culture plate. Moreover, the cloning efficiency of JB6 P+ cells was dramatically decreased by inhibition of p12-LOX. In contrast, baicalein treatment did not affect the cloning efficiency of most malignant cancer cells. These results indicate that p12-LOX is induced by the inflammatory cytokine TNF-alpha in the early stage of tumorigenesis, and is required for tumor promotion through enhancing efficient proliferation of a small number of initiated cells. The present results suggest that the p12-LOX pathway may be an effective target of chemoprevention for skin carcinogenesis.