[Research progress of thyroid hormone in pulmonary fibrosis].

Pulmonary fibrosis is end-stage of variety of heterogeneous interstitial lung disease, characterizedby excessive proliferation of fibroblasts and extracellular matrix deposition and destruction of lung parenchyma. Thyroid and lung are derived from the same endodermal cells, thyroid hormone affect the occurrence、development and prognosis of the chronic obstructive pulmonary disease, lung cancer and other lung diseases, This article reviews the role and mechanism of thyroid hormone in pulmonary fibrosis in order to provide new idea for the study of the role and mechanism of thyroid hormone in silicosis.

Exosome-mediated transfer of MIF confers temozolomide resistance by regulating TIMP3/PI3K/AKT axis in gliomas.

Temozolomide (TMZ) resistance is an important cause of clinical treatment failure and poor prognosis in gliomas. Increasing evidence indicates that cancer-derived exosomes contribute to chemoresistance; however, the specific contribution of glioma-derived exosomes remains unclear. The aim of this study was to explore the role and underlying mechanisms of exosomal macrophage migration inhibitory factor (MIF) on TMZ resistance in gliomas. We first demonstrated that MIF was upregulated in the exosomes of TMZ-resistant cells, engendering the transfer of TMZ resistance to sensitive cells. Our results indicated that exosomal MIF conferred TMZ resistance to sensitive cells through the enhancement of cell proliferation and the repression of cell apoptosis upon TMZ exposure. MIF knockdown enhanced TMZ sensitivity in resistant glioma cells by upregulating Metalloproteinase Inhibitor 3 (TIMP3) and subsequently suppressing the PI3K/AKT signaling pathway. Additionally, exosomal MIF promoted tumor growth and TMZ resistance of glioma cells in vivo, while IOS-1 (MIF inhibitor) promotes glioma TMZ sensitive in vivo. Taken together, our study demonstrated that exosome-mediated transfer of MIF enhanced TMZ resistance in glioma through downregulating TIMP3 and further activating the PI3K/AKT signaling pathway, highlighting a prognostic biomarker and promising therapeutic target for TMZ treatment in gliomas.

[Effect of ectodysplasin-A1 on proliferation and cell cycle of ameloblast-like cell].

Objective: To investigate the effects of ectodysplasin-A1 (EDA1) on the proliferation and cell cycle of ameloblast-like epithelial cells (LS8 cells). Methods: Wild EDA1 plasmid pCR3-Flag-EDA1-W (wild group), syndrome mutant EDA1 plasmid pCR3-Flag-EDA1-H252L (mutant group) and empty vector plasmid pCR3-Flag (control group) were transfected into LS8 cells. Cell proliferation was detected by methyl thiazolyl tetrazolium (MTT) assay and cell cycle was detected by flow cytometry. All tests were repeated three times. Results: Compared with the control group (0.105±0.032), the proliferation activity of the wild group (0.201±0.009) was significantly higher after 72 h (P<0.05). Compared with the control group (0.168±0.054) and the mutant group (0.194±0.059), the proliferation activity of the wild group (0.386±0.066) was significantly higher after 96 h (P<0.05). There was no significant difference between the mutant group and the control group at all time points (P>0.05). In the G0/G1 phase, compared with the control group (65.4%±2.1%) and the mutant group (66.6%±3.1%), the cell distribution ratio of the wild group (51.2%±1.1%) was significantly lower (P<0.01). In the S phase, compared with the control group (23.1%±2.0%) and the mutant group (21.9%±1.8%), the cell distribution ratio of the wild type group (37.3%±2.4%) was significantly higher (P<0.01). There was no significant difference in cell cycle distribution between the mutant group and the control group (P<0.05). Conclusions: Wild EDA1 promotes the proliferation of LS8 cells and the transformation from G0/G1 to S phase. The syndrome mutant EDA1 (EDA1-H252L) loses its function of regulating the cell proliferation and cell cycle of LS8 cells.

Analysis of velopharyngeal function and speech outcomes of Sommerlad palatoplasty combined with sphincter pharyngoplasty in surgical repair of older patients with cleft palate：experience from a major craniofacial surgery centre in eastern China.

We aimed to evaluate velopharyngeal function and speech outcomes of Sommerlad palatoplasty combined with sphincter pharyngoplasty in surgical repair of cleft palate in patients over five years old. Fifty-eight patients were reviewed between the years 2013 and 2017, 31 of whom were treated with Sommerlad palatoplasty combined with sphincter pharyngoplasty, (mean age 15 (range 9 - 22) years), and 27 were treated with Sommerlad palatoplasty alone (mean age 18 (range 10-25) years). Velopharyngeal function was evaluated by radiographic lateral cephalometry and nasoendoscopy. Hypernasality, nasal emissions, and intelligibility were used to assess speech. The rate of velopharyngeal competence was 20/31 in the palatoplasty plus pharyngoplasty group and 7/27 in the palatoplasty alone group after surgical treatment (p=0.003). The improvements in hypernasality (p=0.024), air emission (p=0.004), and speech intelligibility (p=0.004) in the palatoplasty plus pharyngoplasty group was better than that in the palatoplasty alone group. It has been suggested that the surgical approach with the palatoplasty together with the sphincter pharyngoplasty has a higher rate of success in surgical repair of older patients with cleft palate.

[Translational research on gastric cancer: Ruijin experience].

Translational medicine refers to translating basic research achievements into the clinical application for diagnosis and treatment. There are still many issues of gastric cancer to be solved in clinical diagnosis and treatment. We need to focus on clinical questions, by means of basic research and multidisciplinary intersection approach, to further improve the overall efficacy of gastric cancer. Based on Ruijin Hospital's translational research experiences, in this paper, the authors describe the future direction in the field of translational research, such as the etiology and pathogenesis of gastric cancer, tumor markers for early diagnosis, prediction of recurrence and metastasis, classification criteria, evaluation of chemotherapy, tumor heterogeneity, targeted therapy, immunotherapy and research platform establishment. In addition, here we share our perspective of the research on serum biomarkers, specific antibodies and improvement of drug delivery for gastric cancer.

Clinical and genetic analysis of idiopathic normophosphatemic tumoral calcinosis in 19 patients.

PURPOSE: Tumoral calcinosis is a rare clinicopathological entity characterized by ectopic soft-tissue calcification, typically periarticular. Normophosphatemic tumoral calcinosis is seldom reported in East Asian populations, and the preoperative diagnosis is often elusive. This study was performed to characterize the clinical profile of normophosphatemic tumoral calcinosis and investigate the presence of the SAMD9 gene mutation. METHODS: The clinical features, pathological examination findings, and outcomes of 19 subjects were retrospectively reviewed. All patients were analyzed for SAMD9 gene mutation using paraffin-embedded tumoral calcinosis specimens. RESULTS: Nineteen subjects were analyzed (7 males, 12 females). Their mean age at surgery, mean age at symptom onset, and median disease duration was 51.9 ± 17.3 (range 7-75) years, 49.1 ± 17.2 (range 7-74) years, and 1.3 (interquartile range 0.5-3.0) years, respectively. Lesions were located in the hand in 8 (42.1%) subjects; wrist in 5 (26.3%); shoulder in 2 (10.5%); and hip, knee, buttock, and scrotum in 1 (5.3%) subject each. The lesions in 17 (89.5%) subjects were located around the joints [small joints (hand and wrist) in 13 (68.4%) and large joints (shoulder, hip, and knee) in 4 (21.1%)]. Lesions occurred in the upper limbs in 15 (78.9%) subjects and in the lower limbs in 2 (10.5%). Multiple-lesion involvement (distal right index finger and middle finger) occurred in one (5.3%) subject. Symptoms included pain in 15 (78.9%) subjects, impaired mobility in 5 (26.3%), swelling in 5 (26.3%), numbness in 2 (10.5%), and an asymptomatic mass in 2 (10.5%). The serum inorganic phosphorus concentration was normal in all 19 subjects (mean 1.17 ± 0.15 mmol/L). The serum calcium concentration was normal in 18 subjects and low in 1. The serum alkaline phosphatase concentration was normal in all 19 subjects. Pathological examination indicated multiple nodules of calcified materials that manifested an amorphous or granular blue-purple crystal and were surrounded by proliferation of mononuclear or multinuclear macrophages, osteoclastic-like giant cells, fibroblasts, and chronic inflammatory cells. Notably, different phases of pathological manifestations were observed in the same microscopic field. During follow-up (0.5-65.0 months), no recurrence of tumoral calcinosis was observed in 18 (94.7%) subjects, but 1 subject developed in situ recurrence of an asymptomatic subcutaneous mass after 6 months postoperatively. Genetic analysis in all 19 subjects revealed no SAMD9 gene mutations. CONCLUSIONS: Most subjects were females and developed calcinosis in adulthood. Small joints (hand and wrist) and the upper limbs were frequently involved. The presence of different phases of pathological features in the same subject suggests that about half of the study participants had been misdiagnosed with another condition (such as gout, osteoarthritis, etc.). Complete surgical excision led to cure without recurrence during follow-up in majority of the study participants.

[Novel tumor metastasis suppressorgene LASS2/TMSG1 S248A mutant promotes invasion of prostate cancer cells through increasing ATP6V0C expression].

OBJECTIVE: LASS2/TMSG1 gene is a novel tumor metastasis suppressor gene cloned from human prostate cancer cell line PC-3M in 1999 by Department of Pathology,Peking University of Basic Medical Sciences. It was found out that protein encoded by LASS2/TMSG1 could interact with the c subunit of vacuolar-ATPase (ATP6V0C). In this study, we explored the effect of LASS2/TMSG1 and its mutants on proliferation, migration and invasion of human prostate cancer cells and its molecular mechanism. METHODS: We constructed four LASS2/TMSG1 mutants and stably transfected the variants to human prostate cancer cell line PC-3M-1E8 cell with high metastatic potential. The stable transfectants were identified by qPCR and Western blot through analyzing the expression of LASS2/TMSG1 and ATP6V0C, the cell biology functions of LASS2/TMSG1 and its four mutants were studied using growth curve,MTT assay, soft agar colony formation assay, wound migration assay, Matrigel invasion study and flow cytometry. Furthermore, immunofluorescence was used to analysis the interaction of LASS2/ TMSG1 mutants and ATP6V0C. RESULTS: LASS2/TMSG1 mRNA and protein in LASS2/TMSG1 group and Mut1-Mut4 groups were higher than that in Vector group; Western blot showed that ATP6V0C protein in LASS2/TMSG1 wild group was lower than that in Vector group, but ATP6V0C protein in LASS2/TMSG1 S248A group was obviously higher than that in Vector group. MTT test and growth curve assay showed growth ability in LASS2/TMSG1 S248A group was increasing compared with other groups from day 5. Soft Agar colony formation experiment showed anchor independent growth ability in LASS2/TMSG1 S248A group was higher than those in the other groups (P<0.05), Cell migrations (from 35.3%±3.2% to 70.3%±3%) in LASS2/TMSG1 S248A group was increasing compared with LASS2/TMSG1 wild group (P<0.01), and more cells passed through Matrigel in LASS2/TMSG1 S248A group compared with LASS2/TMSG1 wild group (from 50±3.2 to 203±6.5, P<0.01), the apoptosis rate in LASS2/TMSG1 S248A group was obviously higher than that in LASS2/TMSG1 wild group (from 7% to 15.1%, P<0.05), and the G0/G1 ratio in LASS2/TMSG1 S248A group was obviously higher than that in LASS2/TMSG1 wild group (from 51.0% to 85.4%). Furthermore, double immunofluorescent staining observed the colocalization between ATP6V0C and LASS2/TMSG1 protein and its mutations, the expression of ATP6V0C in LASS2/TMSG1 S248A group increased significantly compared with the other groups. CONCLUSION: LASS2/TMSG1 S248A promotes proliferation, migration and invasion of prostate cancer cells through increasing ATP6V0C expression, suggesting that aa248-250 is an important function site for LASS2/TMSG1 in invasion suppression of prostate cancer cells.

MicroRNA-31 inhibits osteosarcoma cell proliferation, migration and invasion by targeting PIK3C2A.

OBJECTIVE: To elucidate the role of microRNA-31 (miR-31) in osteosarcoma and the molecular mechanism of miR-31 in the proliferation, migration, and invasion of osteosarcoma. PATIENTS AND METHODS: Real Time-quantitative Polymerase Chain Reaction (RT-qPCR) was used to examine the expression of microRNA-31 in human osteosarcoma tissues. Pearson's chi-squared test was used to analyze the correlation between microRNA-31 and clinicopathological features. Proliferation, migration, invasion, and PI3K3C2A protein in treated osteosarcoma cells were detected by Cell Counting Kit-8 (CCK-8) assay, transwell assay without Matrigel, transwell assay with Matrigel, and Western blot analysis, respectively. RESULTS: qRT-PCR showed that miR-31 was down-regulated in osteosarcoma tissues compared with paired para-tumor bone tissues. The lower level of miR-31 was closely associated with high-grade osteosarcoma, metastasis, and poor overall survival. CCK-8 and transwell assay showed that miR-31 inhibited osteosarcoma cells proliferation, migration, and invasion. According to luciferase assay, miR-31 inhibits osteosarcoma cell proliferation, migration, and invasion through inhibiting PIK3C2A. Reversely, overexpression of PIK3C2A inhibited partial effect of miR-31 on proliferation, migration, and invasion in vitro. CONCLUSIONS: MiR-31 inhibits osteosarcoma cell proliferation, migration, and invasion by targeting PICK3C2A. MiR-31 can thus be used as a therapeutic target in osteosarcoma treatment.

Preventive effects of low molecular weight heparin on formation of deep vein thrombosis by reducing D dimer values in patients undergoing spinal surgery.

OBJECTIVE: Deep vein thrombosis (DVT) is one of the most common complications for patients undergoing spinal surgery. This study aims to investigate preventive effects of low-molecular-weight heparin (LMWH) on the formation of DVT. PATIENTS AND METHODS: This study involved 37 patients who underwent spinal surgery between April 2016 and April 2017. Patients were divided into LMWH group and Control group. Clinical parameters, including operation time, intra-operative blood loss, incision length, post-operative visual analogue scale (VAS), exercise-time leaving bed and post-operative extubation time, were collected. Blood routine analysis, including platelet count (PLT), red blood cell count (RBC), white blood cell count (WBC) and hemoglobin (HGB) were also conducted. Coagulation parameters, including prothrombin time-international normalized ratio (PT-INR), fibrinogen (FIB), activated partial thromboplastin time (APTT), thrombin time (TT), prothrombin time 1 (PT-1) and D-dimer (DD), were evaluated. The drainage fluid was collected. RESULTS: LMWH treatment significantly decreased operative time, blood loss and incision length compared to Control group at 1st, 3rd and 7th day post-operation (all p<0.05). LMWH treatment significantly increased WBC levels compared to Control group at 1st, 3rd and 7th day post-operation (p<0.05). LMWH treatment significantly decreased DD levels in the post-operative patients compared to Control group at 1st, 3rd and 7th day post-operation (p<0.05). However, LMWH treatment doesn't affect drainage amounts of patients. DD levels were positively correlated with WBC counts for the LMWH treated patients. CONCLUSIONS: Low molecular weight heparin effectively prevents the formation of DVT by reducing DD values in patients undergoing spinal surgery.

[Silencing of vacuolar ATPase c subunit ATP6V0C inhibits invasion of prostate cancer cells].

OBJECTIVE: Vacuolar ATPase (V-ATPase) was found within the membranes and internal organelles of a vast array of eukaryotic cells, and was related to various kinds of highly metastatic tumors. LASS2/TMSG1 gene was a novel tumor metastasis suppressor gene cloned from human prostate cancer cell line PC-3M in 1999 by our laboratory. It was found out that protein encoded by LASS2/TMSG1 could interact with the c subunit of V-ATPase (ATP6V0C). In this study, To use RNA interference to suppress the expression of ATP6V0C and try to further investigate the molecular mechanism of ATP6V0C in tumor metastasis and its relationship with LASS2/TMSG1 gene. METHODS AND RESULTS: The expression level of ATP6V0C mRNA and protein in high metastatic potential prostate cancer cell lines (PC-3M-1E8 and PC-3M) was significantly higher than that in low metastatic potential prostate cancer cell lines (PC-3M-2B4 and PC-3), the expression level in PC-3M-1E8 being the highest. Follow-up tests selected PC-3M-1E8 cells for gene silencing. The expression and secretion of MMP-2 and the expression of MMP-9 in ATP6V0C siRNA transfected PC-3M-1E8 cells displayed no obvious change, but the activity of secreted MMP-9 was abated noticeably compared with the controls (P<0.05). Extracellular hydrogen ion concentration and V-ATPase activity in interference group were both reduced significantly compared with the controls (P<0.05). The migration and invasion capacity of ATP6V0C siRNA interfered cells in vitro were diminished significantly compared with the controls (P<0.05). Furthermore, a dramatic reduction of LASS2/TMSG1 mRNA and protein level after transfection of siRNA in PC-3M-1E8 cells was discovered (P<0.05). Confocal immunofluorescence showed a vast co-localization of ATP6V0C protein and LASS2/TMSG1 protein in plasma and membrane. The co-localization signals of control group were much stronger than those of interference group. CONCLUSION: Specific siRNA silencing of ATP6V0C gene inhi-bits the invasion of human prostate cancer cells in vitro by mechanism of inhibiting V-ATPase activity and then reducing the extracellular hydrogen ion concentration, inhibiting MMP-9 activation and affecting ECM degradation and reconstruction. Meanwhile, ATP6V0C and LASS2/TMSG1 have interaction and it is likely that ATP6V0C functions as a feedback regulator of LASS2/TMSG1.

MiR-145 silencing promotes steroid-induced avascular necrosis of the femoral head repair via upregulating VEGF.

OBJECTIVE: To investigate the role of miR-145 silencing in the expression of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), B-cell lymphoma-2 (Bcl-2), BCL2-Associated X(bax) and caspase-3 in avascular necrosis of femoral head (ANFH). MATERIALS AND METHODS: A total of 12 healthy wild-types (the control group) and 12 miR-145 knock-out (miR-145-/-) (the experimental group) adult New Zealand white rabbits were selected to construct ANFH model with a steroid. Four weeks later, immunohistochemistry, qRT-PCR and Western blot were performed to measure the VEGF, bFGF, Bcl-2, bax, caspase-3, ß-catenin as well as c-Myc expression. Terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL) staining analysis was used to detect the apoptosis of bone cells in each group. RESULTS: Compared with the control group, the expression of VEGF, bFGF, Bcl-2, ß-catenin and c-Myc in the miR-145-/- group raised (p<0.05). Moreover, the expression level of bax and caspase-3 significantly decreased in the miR-145-/- group (p<0.05). TUNEL staining showed decreased apoptosis in the miR-145-/- group. CONCLUSIONS: MiR-145 silencing promotes bone repair of ANFH via upregulating VEGF, bFGF and inhibiting the bone cells apoptosis through Wnt/ß-catenin pathway.

Enhanced toxicity to the cyanobacterium Microcystis aeruginosa by low-dosage repeated exposure to the allelochemical N-phenyl-1-naphthylamine.

It has been puzzling whether and how a plant could exert a strong allelopathic inhibition to the target organisms by releasing low concentrations of allelochemicals. Plant allelochemicals have been proposed to be released continuously, however, direct evidence from specific allelochemicals is urgently required. In the present study, the toxicity of allelochemical N-phenyl-1-naphthylamine (NPN) towards the cyanobacterium Microcystis aeruginosa by two different exposure patterns was compared. One was low-dosage repeated exposure (LRE), in which 50  µg L-1 NPN was repeatedly dosed to simulate the continual release of allelochemicals, and the other one was high-dosage single exposure (HSE) as per the routine toxicity assay. The results showed a significant growth inhibition to M. aeruginosa in the LRE group, where the inhibition rate reached above 90% from day 6 to day 9. The cell-membrane damage ratio increased from 64.05% on day 5 up to 96.60% on day 9. PSII photosynthesis activity expressed as Fv/Fm, ΦPSII, NPQ and ETRmax was also thoroughly inhibited in this group. Whereas the growth and PSII photosynthesis activity of M. aeruginosa in the HSE group were inhibited initially, but recovered gradually from day 4 or 5, which was accompanied by a continuous reduction of NPN content in culture solutions. Although NPN content in the LRE group was relatively lower, it remained at a more stable level throughout the experiment. These results indicate that continual release of low-dosage allelochemicals by aquatic plants plays crucial roles in their potent inhibition against cyanobacteria. Low-dosage continual exposure pattern needs to be investigated further.

Post-surgical treatment of a patient with ectopic pheochromocytoma.

We report here the course and treatment of a patient with ectopic pheochromocytoma. The patient was cured after treatment with respiratory and circulatory support, multiple-organ protection, and continuous renal replacement therapy (CRRT) for approximately 2 weeks. After misdiagnosis, a patient with ectopic pheochromocytoma who is being treated should undergo aggressive fluid supplementation and CRRT instead of central venous pressure measurement, which has limited value in guiding fluid supplementation. The main priority is maintaining hypotension. Hypertension may be controlled with rapid-acting agents. A good outcome can be anticipated for patients who undergo comprehensive intensive care unit therapy.

Upregulated Hoxc6 expression is associated with poor survival in gastric cancer patients.

Human Hox genes (Homeobox) have crucial roles in development and differentiation, regulating numerous processes including apoptosis, receptor signalling, differentiation, motility and angiogenesis. Aberrant expression of Hoxc6 gene has been reported in several tumor tissues and cancer cell lines. The prognostic significance of Hoxc6 in gastric cancer remains largely unknown.This study was aimed to investigate the clinical significance of Hoxc6 in gastric cancer.Total RNA of paired tissue samples (n=25) and a tissue microarray containing 161 paired tissues from patients with gastric cancers at different stages were collected. Quantitative real-time PCR and immunochemistry assay were carried out to investigate the expression of Hoxc6. Hoxc6 mRNA was increased in gastric cancer tissues ( 16 of 25) compared with the adjacent normal mucosa (P<0.05). Immunohistochemical detection showed that expression of Hoxc6 was associated with the depth of tumor invasion (P<0.05). Patients with higher expression levels of Hoxc6 had a shorter overall survival rate (P<0.05).Hoxc6 might contribute to the progression of gastric carcinogenesis and may be a significant predictor of poor survival in patients with gastric cancer after curative operations.

Clinicopathological characteristics and computed tomography features of mucinous gastric carcinoma.

This study investigated the clinicopathological characteristics of mucinous gastric carcinoma (MGC) and assessed whether multidetector-row computed tomography (MDCT) could differentiate MGC from non-mucinous gastric carcinoma (NGC). Clinicopathological data from 542 patients with gastric carcinoma (23 MGC, 519 NGC), who underwent pre-operative MDCT examination and curative or palliative gastrectomy, were analysed. Only seven of the 23 patients with MGC were correctly diagnosed pre-operatively by endoscopic biopsy. The MGC patients had larger tumours, a higher frequency of lymph node metastases, were more likely to have tumours of tumour, node, metastasis stages III and IV, and were less likely to have a curative resection than NGC patients. In addition, five MGC patients had calcifications in the thickened gastric wall. In conclusion, MGC is rare and is detected mostly at an advanced stage. The diagnostic sensitivity of MGC by endoscopic biopsy was relatively low, whereas MDCT was helpful in distinguishing MGC from NGC.

Size of the largest lymph node visualized on multi-detector-row computed tomography (MDCT) is useful in predicting metastatic lymph node status of gastric cancer.

This study was designed to investigate whether the size of the largest lymph node (long-axis diameter [LAD] and short-axis diameter [SAD]) visualized using multi-detector-row computed tomography (MDCT) was useful for predicting the metastatic lymph node (MLN) status of gastric cancer. A retrospective analysis of 305 gastric cancer patients who underwent pre-operative MDCT was performed, followed by a prospective study in 61 gastric cancer patients to determine the diagnostic effectiveness of LAD and SAD. In the retrospective study, the accuracy of LAD and SAD for predicting the MLN status of gastric cancer was 51.1% and 45.9%, respectively. In the prospective study, the accuracy of LAD and SAD measurement and the traditional MDCT method of counting MLNs was 52.5%, 49.2% and 57.4%, respectively; the differences were not significant. In conclusion, the size of the largest lymph node in terms of LAD and SAD visualized on MDCT was useful for predicting the MLN status of gastric cancer, with accuracy comparable to the traditional MDCT method of counting the total number of MLNs detected.

Non-calcifying variant of calcifying epithelial odontogenic tumor with Langerhans cells.

Calcifying epithelial odontogenic tumor (CEOT) is a rare type of odontogenic tumor. The most characteristic feature of the classical CEOT is the presence of amyloid globules and Liesegang ring calcification in the tumor tissue. Here, we present a non-calcifying variant of intraosseous CEOT with the presence of Langerhans cells within tumor epithelial nests in a 52-year-old Taiwanese woman. The patient was referred from a local dentist to our hospital for treatment of a unilocular radiolucent lesion at the right anterior region of the maxilla. The lesion was excised. Microscopically, the tumor was composed of small nests or strands of odontogenic epithelial cells and amorphous eosinophilic globules of amyloid-like materials in a loose fibrous connective tissue stroma. The tumor epithelial cells were positive for pan-cytokeratins (AE1 and AE3). Langerhans cells demonstrated by anti-CD1a staining were found in nests or strands of tumor epithelial cells. The eosinophilic globules were positive for Congo red and showed green birefringence when subjected to polarized light. Review of the English literature revealed two cases of non-calcifying variant of intraosseous CEOT with Langerhans cells in the anterior and premolar regions of the maxilla. Taken together, we suggest that the non-calcifying, Langerhan cell-rich variant of CEOT may have a distinct predilection for occurrence in the anterior and premolar region of the maxilla in contrast to the classical CEOTs that usually occur in the molar and ascending ramus area of the mandible.

Effect of NF-kappaB constitutive activation on proliferation and apoptosis of gastric cancer cell lines.

OBJECTIVE: To observe whether there is constitutive activation of nuclear transcription factor kappaB (NF-kappaB) and its effect on proliferation and apoptosis of human gastric cancer cell lines. METHODS: Nuclear/cytoplasmic protein expression of NF-kappaB was analyzed by Western blot in four different gastric cancer cell lines. Trans AM(TM) NF-kappaB p65 Kit was used for detecting the difference of p65 activity. The effect of PDTC (pyrrolidine dithiocarbamate), a specific inhibitor of NF-kappaB on the proliferation of gastric cancer cells, was measured by MTT (3-[4,5-dimethythiazol-2-yl]-2,5-diphenyltetrazolium bromide) method. The apoptotic rates of AGS and SGC-7901 gastric cancer cell lines were measured with flow cytometer (FCM) after treatment by PDTC. RESULTS: The constitutive activations of NF-kappaB were identified in four gastric cancer cell lines. The expression of activated subunit of p50 was lower in AGS cell line, and higher in MKN28, MKN45 and SGC-7901 cell lines. The expression of activated subunit of p65 was lower in MKN28 and MKN45 cell lines, and higher in AGS and SGC-7901 cell lines. Both the activity of NF-kappaB and the cell proliferation were significantly inhibited in experimental group treated by PDTC, compared with control groups (p<0.01). An increased apoptotic rate and a decreased proliferating activity were observed after the gastric cancer cells were exposed to PDTC for 24 h. CONCLUSIONS: These results suggested that the constitutive activation and the protein expression of NF-kappaB are different in gastric cancer cell lines. PDTC can inhibit NF-kappaB activity and cell proliferation, which related to an increased cell apoptosis. The results disclosed that NF-kappaB could be a potential therapeutic target for solid tumor therapy.

Cyclooxygenase-2 expression is associated with VEGF-C and lymph node metastases in gastric cancer patients.

Previous studies have suggested that cyclooxygenase-2 (COX-2) over-expression is associated with angiogenesis in gastric cancer. However, the relationship between COX-2 and lymphangiogenesis is still unclear. The aim of this study was to determine the relationship between COX-2 expression and lymphangiogenic factor, vascular endothelial growth factor-C (VEGF-C), in human gastric cancer, as well as to correlate with clinicopathological parameters. Sixty-three gastric cancer patients underwent radical gastrectomy (D2 or D3) were enrolled in this study. The expression of COX-2 and VEGF-C were detected by immunohistochemistry, and the small lymphatic vessels were immunohistochemically stained by LYVE-1 antibody. The association between COX-2 and VEGF-C expressions and clinicopathological parameters (such as gender, tumor location, lymph node status and Lauren classification) were determined. VEGF-C over-expression was observed in 33 of 63 patients (52%), while COX-2 over-expression occurred in 42 of 63 tumor samples (67%). Presence of microlymphatic vessels with LYVE-1 staining was found in 35 cases. COX-2 over-expression was highly correlated with VEGF-C over-expression (P = 0.032), microlymphatic vessels (P = 0.002) as well as presence of metastatic lymph nodes (P = 0.007). However, no significant correlation was found between COX-2 expression and other clinicopathological parameters. Our data suggest that COX-2 expression is associated with lymphangiogenesis and lymph node metastasis in human gastric carcinoma. This raises the possibility that COX-2-mediated VEGF-C over-expression might promote lymph node metastasis via lymphangiogenesis pathway in patients with gastric cancer.

Germline mutational analysis of CDH1 and pathologic features in familial cancer syndrome with diffuse gastric cancer/breast cancer proband in a Chinese family.

AIMS: Hereditary non-polyposis colorectal cancer, thyroid medullary carcinoma, breast/ovarian cancer and gastric cancer/breast cancer syndrome are encountered in surgery. Some gastric cancer/breast cancer syndrome may be the result of a CDH1 germline mutation. This is the first report of CDH1 germline mutations gastric cancer/breast cancer syndrome in Chinese patients. METHODS: Peripheral blood from the proband, as well as, her first and second degree relatives was collected and CDH1 gene exon 1-16 mutations were screened. E-cadherin/beta-catenin proteins expression and histopathologic features were examined on gastric cancer/breast cancer tissues from the proband. RESULTS: A C-->T nucleotide substitution at exon 13 (mRNA 2200 locus, Accession number NM-004360) was found. This was a transition from GCC-->GCT in DNA sequence (Ala154Ala). Diffuse-type gastric cancer and infiltrating ductal breast carcinoma were present. Both tumours preserved E-cadherin/beta-catenin expression immunohistochemically. CONCLUSIONS: Familial cancer syndrome with diffuse-type gastric cancer/breast cancer proband in Chinese has a propensity of early onset during lifespan. No truncating or splice-site CDH1 mutations had been identified in this family. A silent nucleotide variation in exon 13 of the CDH1 gene may contribute to some forms of cancer susceptibility.