OTX1 Contributes to Hepatocellular Carcinoma Progression by Regulation of ERK/MAPK Pathway.

Orthodenticlehomeobox 1 (OTX1) overexpression had previously been associated with the progression of several tumors. The present study aimed to determine the expression and role of OTX1 in human hepatocellular carcinoma (HCC). The expression level of OTX1 was examined by quantitative real-time PCR (qRT-PCR) in 10 samples of HCC and paired adjacent non-cancerous tissues, and by immunohistochemistry (IHC) analysis in 128 HCC samples and matched controls. The relationship between OTX1 expression and the clinicopathological features werealso analyzed. Furthermore, the effects of OTX1 knockdown on cell proliferation and migration were determined in HCC cell lines. Axenograft mouse model was also established to investigate the role of OTX1 in HCC tumor growth. TheqRT-PCR and IHC analyses revealed that OTX1 was significantly elevated in HCC tissues compared with the paired non-cancerous controls. Expression of OTX1 was positively correlated with nodal metastasis status (P = 0.009) and TNM staging (P = 0.001) in HCC tissues. In addition, knockdown of OTX1 by shRNA significantly inhibited the proliferation and migration, and induced cell cycle arrest in S phase in vitro. Tumor growth was markedly inhibited by OTX1 silencing in the xenograft. Moreover, OTX1 silencing was causable for the decreased phosphorylation level of ERK/MAPK signaling. In conclusion, OTX1 contributes to HCC progression possibly by regulation of ERK/MAPK pathway. OTX1 may be a novel target for molecular therapy towards HCC.

The effect of Chinese Jinzhida recipe on the hippocampus in a rat model of diabetes-associated cognitive decline.

BACKGROUND: To investigate the effects of treatment with Multi component Chinese Medicine Jinzhida (JZD) on behavioral deficits in diabetes-associated cognitive decline (DACD) rats and verify our hypothesis that JZD treatment improves cognitive function by suppressing the endoplasmic reticulum stress (ERS) and improving insulin signaling transduction in the rats' hippocampus. METHODS: A rat model of type 2 diabetes mellitus (T2DM) was established using high fat diet and streptozotocin (30 mg/kg, ip). Insulin sensitivity was evaluated by the oral glucose tolerance test and the insulin tolerance test. After 7 weeks, the T2DM rats were treated with JZD. The step-down test and Morris water maze were used to evaluate behavior in T2DM rats after 5 weeks of treatment with JZD. Levels of phosphorylated proteins involved in the ERS and in insulin signaling transduction pathways were assessed by Western blot for T2DM rats' hippocampus. RESULTS: Compared to healthy control rats, T2DM rats initially showed insulin resistance and had declines in acquisition and retrieval processes in the step-down test and in spatial memory in the Morris water maze after 12 weeks. Performance on both the step-down test and Morris water maze tasks improved after JZD treatment. In T2DM rats, the ERS was activated, and then inhibited the insulin signal transduction pathways through the Jun NH2-terminal kinases (JNK) mediated. JZD treatment suppressed the ERS, increased insulin signal transduction, and improved insulin resistance in the rats' hippocampus. CONCLUSIONS: Treatment with JZD improved cognitive function in the T2DM rat model. The possible mechanism for DACD was related with ERS inducing the insulin signal transduction dysfunction in T2DM rats' hippocampus. The JZD could reduce ERS and improve insulin signal transduction and insulin resistance in T2DM rats' hippocampus and as a result improved the cognitive function.

Cell viability and drug evaluation biosensing system based on disposable AuNPs/MWCNT nanocomposite modified screen-printed electrode for exocytosis dopamine detection.

Cell viability, as an important index to evaluate drug effects, usually was measured by tetrazolium colorimetric assay, playing a key role in drug development and drug screening. Tedious operating procedures, unsatisfactory sensitivity and abominable environments perplex researchers to acquire more detailed in vivo-relevant biological information. Herein, a simple and low-cost cell viability and drug evaluation biosensing system-based on multiwalled carbon nanotubes, gold nanoparticles and Nafion modified screen-printed electrode (SPE) biosensor was constructed for detection of dopamine (DA) released from living cells to evaluate cytotoxicity of antineoplastic drugs such as cisplatin and resveratrol. The biosensing system was demonstrated to display exceptional selectivity, excellent flexibility and good stability toward DA measurement in complex bio-samples. Additionally, the satisfactory recoveries of DA in real samples revealed the reliability and accuracy of the biosensing system in practical application. The IC50 curves respectively obtained by the biosensing system and tetrazolium colorimetric assay provided similar IC50 value but distinctly different dose-effect relationship, which confirmed the enormous potential of the biosensor in cell viability and described drug efficacy profiles in cell function. In short, the cell viability and drug evaluation system using SPE biosensor paves a new way in drug screening and pharmaceutical application to measure bioactive molecule such as DA.

[Diagnosis and therapy on hemangiomas and vascular malformation in view of the new classification].

Vascular birthmarks are the most common disease. The morbidity is about 2.5%, most of the lesions occur in oral and maxillofacial regions which accounts for 40%-60% of the total lesions. In 1982, Mulliken and Glowacki proposed a biologic classification of vascular birthmarks on the basis of their clinical manifestations, histopathological features, and natural history. They defined hemangiomas as vascular tumors with a growth phase, marked by endothelial proliferation and hypercellularity, and an involutional phase. They recognized that many entities referred to as hemangiomas are actually structural malformations of the vasculature, derived from capillaries, veins, lymph vessels, or arteries or from a combination of these sources. The classification was confirmed and issued by International Society for the study of vascular anomaly (ISSVA) in 1988. Waner and Suen amended the above category in 1995. This paper presents the new classification of vascular birthmarks and the developments in this field in recent years, including the pathology, clinical features and the therapy. For example, the classification of venular malformation categorized by Waner in 1989; the classification of lymphous malformation by Waner and Suen in 1995; and the treatments according to above classifications.

Prevalence And Clinical Significance Of Oncogenic CD79B And MYD88 Mutations In Primary Testicular Diffuse Large B-Cell Lymphoma: A Retrospective Study In China.

PURPOSE: In this study, we investigated the prevalence of CD79B and MYD88 mutations and their relation to clinical characteristics in a cohort of Chinese patients with primary testicular diffuse large B cell lymphoma (PT-DLBCL). PATIENTS AND METHODS: We examined the mutational status of CD79B and MYD88 by Sanger sequencing, and the gene amplification and protein expression of MYD88 in tissue samples from 30 cases of PT-DLBCL by quantitative polymerase chain reaction and immunohistochemistry, respectively. Western blotting was used to analyze phosphorylated STAT3 (p-STAT3) and phosphorylated p65 (p-p65) protein expression in cell lines harboring retroviral constructs for WT MYD88 or MYD88 mutant. RESULTS: Immunophenotypically, MYD88 protein staining was positive in 26/30 (86.67%) cases, and 23/30 (76.7%) cases tested positive for p65 in the nucleus. Genetically, CD79B mutation was found in 13/30 (43.3%) cases, whereas the MYD88 L265P mutation was found in 18/30 (60.0%) cases. Interestingly, CD79B and MYD88 mutations were more prevalent in the non-germinal center B cell (GCB) subtype (83.3% and 76.9%, respectively) and were relatively rare in the GCB subtype (16.7% and 23.1%, respectively). Furthermore, although MYD88 was significantly amplified in PT-DLBCL, the amplification status showed no correlation with its mutational status and protein expression. Clinicopathological comparison between the mutant and wild-type group showed that both CD79B mutation and MYD88 L265P were not significantly correlated with age, anatomical site, Ann Arbor stage, non-GCB/GCB subtype, p65 protein expression, BCL-2 protein expression, or BCL-2/c-MYC double expression (P>0.05). Survival analyses showed that high IPI and advanced stage (stage III-IV) associated with worse outcome (P<0.05). The expression of p-STAT3 and p-p65 protein was upregulated in the mutant group, indicating that MYD88 mutant activated NF-κB and JAK-STAT3 signaling. CONCLUSION: Our results suggest that MYD88 and CD79B mutations are important drivers of immune-privileged site-associated DLBCL and highlight potential therapeutic targets for personalized treatment.

DDR2 facilitates papillary thyroid carcinoma epithelial mesenchymal transition by activating ERK2/Snail1 pathway.

The upregulation of discoidin domain receptor tyrosine kinase 2 (DDR2) has been reported to be associated with poor prognosis and metastasis in numerous tumor types by inducing epithelial-mesenchymal transition (EMT); however, the expression profile of DDR2 in papillary thyroid carcinoma (PTC) with local metastasis and the effect of DDR2 on PTC cells remain unknown. The aim of the present study was to investigate the expression levels of DDR2 in tumor tissues of patients with PTC with local metastasis and cell lines and to determine the effect of DDR2 on EMT in PTC cells. In the present study, it was demonstrated that DDR2 was significantly increased in tumor tissues of patients with PTC with local metastasis and human PTC cell lines. The overexpression of DDR2 by lentiviral transfection decreased E-cadherin protein, increased Vimentin protein, and promoted cell migration and invasion. The inhibition of DDR2 reversed transforming growth factor-ß- and collagen I-induced EMT. EMT induced by DDR2 overexpression was suggested to be dependent on increased Snail1 protein level following extracellular signal-regulated kinase (ERK)2 activation. The inhibition of Snail1 or ERK2 was sufficient to abrogate DDR2-induced PTC cell EMT. In conclusion, these results indicate that DDR2 is upregulated in PTC tissues with local metastasis. Overexpression of DDR2 induced EMT in PTC cells by activating ERK2 and stabilizing Snail1, making it a promising therapeutic target for reducing PTC local or distant metastasis.

Comparison of the c-MET gene amplification between primary tumor and metastatic lymph nodes in non-small cell lung cancer.

BACKGROUND: c-MET has recently been identified as a promising novel target in non-small cell lung cancer (NSCLC). We detected the consistency of c-MET gene amplification in metastatic lymph nodes and tumor tissues of NSCLC patients and discuss the clinical application value of c-MET gene amplification in metastatic lymph nodes. METHODS: Real-time fluorescent quantitative PCR was used to test tumor tissues in 368 NSCLC patients and 178 paired metastatic lymph node samples. The amplification consistency in metastatic lymph nodes and tissue samples were compared and the correlation between c-MET gene amplification and the clinical characteristics of patients was analyzed. RESULTS: The c-MET gene amplification rate was 8.97% (33/368) in tumor tissues. Of the 178 paired cases, c-MET gene amplification was positive in 7.95% (15/178) of cancerous tissues and 18.54% (33/178) of metastatic lymph nodes. c-MET gene amplification was detected more frequently in metastatic lymph nodes than in primary cancerous tissue. When metastatic lymph nodes were used as surrogate samples of primary cancerous tissues, the sensitivity was 86.67% (13/15) and the specificity was 87.69% (143/163). CONCLUSIONS: Screening for c-MET gene amplification in lymph node metastases could determine which patients are eligible for tyrosine kinase inhibitor therapy. Lymph node metastasis can predict c-MET gene amplification in a primary tumor and guide the clinical use of c-MET gene targeted drugs.

[Synchronous primary cancer of the endometrium and ovary].

UNLABELLED: To investigate the clinical and pathological characteristics, treatment, and The data of 12 patients prognosis of synchronous primary cancer of the endometrium and ovary. Methods with synchronous primary cancer of the endometrium and ovary were retrospectively reviewed . Results Eight patients had the same histological type of endometrioid carcinoma in both uterus and ovary, 4 patients had different histological types in uterus and ovary. Synchronous primary cancer of the endometrium and ovary was difficult to be dignosed preoperatively. All ovarian tumors were small with an average diameter of 7 cm. Infertility was common among these patients(40.7%). Most of them had early stage I lesion (66.7%). endometrioid carcinomas was the main pathologic type (66.7%). All patients were treated surgically followed by chemotherapy with a 3-year survival rate of 66.7% (8/12). CONCLUSION: Synchronous primary endometrium and ovary cancer is a specific kind of tumor different from either the primary endometrium carcinoma or ovary carcinoma, and usually can be detected in early stage with a good prognosis.

Anti-inflammatory and hepatoprotective effects of total flavonoid C-glycosides from Abrus mollis extracts.

The aim of this study was to evaluate the anti-inflammatory and hepatoprotective effects of the total flavonoid C-glycosides isolated from Abrus mollis extracts (AME). In the anti-inflammatory tests, xylene-induced ear edema model in mice and carrageenan-induced paw edema model in rats were applied. The hepatoprotective effects of AME were evaluated with various in vivo models of acute and chronic liver injury, including carbon tetrachloride (CCl4)-induced hepatitis in mice, D-galactosamine (D-GalN)-induced hepatitis in rats, as well as CCl4-induced hepatic fibrosis in rats. In the acute inflammation experiment, AME significantly suppressed xylene-induced ear edema and carrageenan-induced paw edema, respectively. In the acute hepatitis tests, AME significantly attenuated the excessive release of ALT and AST induced by CCl4 and D-GalN. In CCl4-induced hepatic fibrosis model, AME alleviated liver injury induced by CCl4 shown by histopathological sections of livers and improved liver function as indicated by decreased liver index, serum ALT, AST, TBIL, and ALP levels and hydroxyproline contents in liver tissues, and increased serum ALB and GLU levels. These results indicated that AME possesses potent anti-inflammatory activity in acute inflammation models and hepatoprotective activity in both acute and chronic liver injury models. In conclusion, AME is a potential anti-inflammatory and hepatoprotective agent and a viable candidate for treating inflammation, hepatitis, and hepatic fibrosis.

Protective effect of total flavonoid C-glycosides from Abrus mollis extract on lipopolysaccharide-induced lipotoxicity in mice.

Abrus mollis is a widely used traditional Chinese medicine for treating acute and chronic hepatitis, steatosis, and fibrosis. It was found that the total flavonoid C-glycosides from Abrus mollis extract (AME) showed potent antioxidant, anti-inflammatory, and hepatoprotective activities. To further investigate the hepatoprotective effect of AME and its possible mechanisms, lipopolysaccharide (LPS)-induced liver injury models were applied in the current study. The results indicated that AME significantly attenuated LPS-induced lipid accumulation in mouse primary hepatocytes as measured by triglyceride (TG) and total cholesterol (TC) assays and Oil Red O staining. Meanwhile, AME exerted a protective effect on LPS-induced liver injury as shown by decreased liver index, serum aminotransferase levels, and hepatic lipid accumulation. Real-time PCR and immunoblot data suggested that AME reversed the LPS-mediated lipid metabolism gene expression, such as sterol regulatory element-binding protein-1 (SREBP-1), fatty acid synthase (FAS), and acetyl-CoA carboxylase 1 (ACC1). In addition, LPS-induced overexpression of activating transcription factor 4 (ATF4), X-box-binding protein-1 (XBP-1), and C/EBP homologous protein (CHOP) were dramatically reversed by AME. Furthermore, AME also decreased the expression of LPS-enhanced interleukin-6 (IL-6) and cyclooxygenase-2 (COX-2). Here, it is demonstrated for the first time that AME ameliorated LPS-induced hepatic lipid accumulation and that this effect of AME can be attributed to its modulation of hepatic de novo fatty acid synthesis. This study also suggested that the hepatoprotective effect of AME may be related to its down-regulation of unfolded protein response (UPR) activation.

[Clinicopathological features and prognostic factors of 216 cases with primary gastrointestinal tract non-Hodgkin's lymphoma].

OBJECTIVE: To investigate the clinicopathological features of primary gastrointestinal non-Hodgkin's lymphomas (PGI-NHL) and their prognostic values. METHODS: The clinical and pathological data of 216 patients diagnosed as PGI-NHL from Zhejiang Cancer Hospital were analyzed retrospectively. χ² test, log-liner model analysis, COX proportional hazard regression analysis and Life-table survival analysis were used to analyze the survival status of the patients by SAS 8.2 software, and Log-rank test was performed to couple the overall survival rates with different prognostic factors. RESULTS: Totally, the age of onset was 8 to 89 years with the median age of 56.5 years. Male versus female was 1.27∶1(121∶95). The most frequently involved location was stomach (147 cases, 68.1%), followed by ileocecus (25 cases, 11.6%), large intestine (20 cases, 9.3%), small intestine (17 cases, 7.9%) and multiple GI involvement (5 cases, 2.3%). 182 cases were classified as B cell lymphomas, 22 cases as T cell lymphomas, and 12 cases not classified exactly due to insufficient data. The 3-year and 5-year survival rates of the patients were 69.4% and 53.3%, respectively. Univariate analysis revealed that age＞60 years, ECOG≥2, high LDH level, stage Ⅲ-Ⅳ, IPI≥2, T cell type and intestinal involvement were predictors for poor prognosis. IPI≥2, T cell type and intestinal involvement were independent adverse predictors for prognosis by multiple COX proportional hazard regression analysis. Among different treatment groups, cases received chemotherapy combined with local radiotherapy gained the best survival status. CONCLUSION: B-cell lymphoma was the main pathological type in PGI-NHL; IPI≥2, T-cell type and intestinal involvement are independent adverse predictors for prognosis; chemotherapy combined with local radiotherapy might be the choice of approach for advanced stage and aggressive PGI-HNL.

[Primary oncocytic carcinoma of the salivary glands: a clinical and pathologic study of 12 cases].

PURPOSE: To investigate the clinicopathological feature of oncocytic carcinoma of the salivary gland, and discuss diagnosis, treatment and prognosis. METHODS: From March 2001 to September 2010, the clinical data and pathological features of 12 cases of oncocytic carcinoma of the salivary gland in Zhejiang Cancer Hospital were reviewed and analyzed retrospectively. All cases are followed up. The Kaplan-Meier method was used to calculate survival curves by SPSS 16.0 software package. RESULTS: The tumors were found mainly as a painless, irregular-shaped mass or lymphadenectasis in the head and neck firstly. Pathologically, oncocytic carcinoma of salivary gland origin was an extremely rare proliferation of malignant oncocytes with adenocarcinomatous architectural phenotypes, including prominent nucleoli and infiltrative qualities. Surgery was the principal treatment, and postoperative radiotherapy was used as adjuvant treatment. Of the 12 cases with follow-up for 6 to 120 months, 7 cases survived without regional or distant metastases. 1 case survived with regional and distant metastases. 2 cases died of regional recurrences.1 case had lymphatic metastasis and died of distant metastasis finally.1 case had given up therapy and died of tumor progress ultimately. 3 cases had local recurrence within 2 years, and the recurrence rate was 25%; 3 cases died within 2 years, and the mortality rate was 25%. CONCLUSIONS: Oncocytic carcinoma of salivary gland origin is an extremely rare tumor in head and neck, with short course and rapid progress. Radical resection postoperative radiotherapy is the treatment of choice. The prognosis of oncocytic carcinoma may be associated with tumor stage, regional lymph node metastases and complete surgical excision.