Treatment with ginkgo biloba extract protects rats against acute pancreatitis-associated lung injury by modulating alveolar macrophage.

INTRODUCTION: Acute pancreatitis (AP) protease release induces lung parenchymal destruction via inflammatory mediators. Ginkgo biloba has been reported to have anti-inflammatory effects. AIM: To evaluate the effect of ginkgo biloba extract on experimental acute pancreatitis-associated lung injury in the rat and to investigate the underlying mechanisms. MATERIAL AND METHODS: Acute pancreatitis was induced in rats by injection of 5% sodium taurocholate into the biliary pancreatic duct. Ginkgo biloba extract (GBE) was administered and pancreas and lung injury were assessed by histological examination. Alveolar macrophages were harvested by bronchoalveolar lavage. Specificity fluorescent probe DAF-FM-DA was applied to observe nitric oxide (NO) bioavailability in alveolar macrophage. The expression of tumour necrosis factor α (TNF-α) and macrophage migration inhibitory factor (MIF) protein in alveolar macrophage was studied by ELISA. RESULTS: In sodium taurocholate-induced acute pancreatitis, treatment with GBE significantly protected rats against lung injury associated with pancreatitis in histological sections. Ginkgo biloba extract had a tendency to down-regulate NO bioavailability compared with the AP group, but without statistical significance. Moreover, TNF-α and MIF at protein levels in alveolar macrophage with GBE treatment were decreased compared with the AP group. CONCLUSIONS: These results suggest that GBE could effectively protect rats against acute pancreatitis-associated lung injury. The GBE may inhibit excessive activation of alveolar macrophages from acute pancreatitis-associated lung injury through down-regulation of generation of NO, TNF-α and MIF. These findings suggest that ginkgo biloba extract is a suitable candidate as an effective strategy against acute pancreatitis-associated lung injury.

Expression and significance of angiopoietin-2 and cyclin D1 in laryngeal squamous cell carcinoma and the correlation with prognosis.

The aim of this study was to study the expression of angiopoietin-2 (Ang-2) and the cell cycle protein D1 (cyclin D1) in laryngeal squamous cell carcinoma (SCC), and its clinicopathological meaning. The expression of Ang-2 and cyclin D1 was analyzed in 116 cases of laryngeal SCC, five cases of atypical hyperplasia and five cases of vocal polyp tissues using the immunohistochemical streptavidin-perosidase (S-P) method. Ang-2 and cyclin D1 protein expression was not present in 5 cases of atypical hyperplasia or 5 cases of vocal cord polyps. However, the proportions of positive staining in well, moderately and poorly differentiated laryngeal SCC were 40, 66.7 and 100%, respectively, for Ang-2 and 50, 66.7, 100%, respectively, for cyclin D1, and were statistically significant (P<0.05). The expression of Ang-2 was positively correlated with cyclin D1 in the laryngeal SCC (r=0.5042; P<0.05). This showed that the tumor grading and cyclin D1 were independent factors affecting laryngeal SCC patient survival by the Cox regression model of risk factors proportion analysis. Ang-2 is synergistic with cyclin D1 in the development of laryngeal SCC. Ang-2 is associated with the metastasis of lymph nodes. Detection of both Ang-2 and cyclin D1 may possess clinical significance in evaluating the prognosis and guiding the clinical treatment of SCC.

Targeting of colorectal cancer growth, metastasis, and anti-apoptosis in BALB/c nude mice via APRIL siRNA.

A proliferation-inducing ligand (APRIL) is overexpressed in most tumor cells and tissues, especially in tumors of the alimentary system, such as colorectal cancer (CRC), gastric cancer, and liver cancer. RNA interference (RNAi) has been proved to be a powerful tool for gene knockdown and holds great promise for the treatment of cancer. In this study, the efficacy of RNAi targeting APRIL was analyzed via relevant experiments on human CRC xenografted in BALB/c nude mice. Both the mRNA and protein levels of APRIL were examined after intratumoral injection of APRIL small interfering RNA (siRNA). Meanwhile, pathological tools were utilized to observe the alterations on the aspects of proliferation, metastasis, apoptosis and cellular necrosis by means of detecting proliferating cell nuclear antigen, Ki-67, MMP-2, MMP-9, TIMP-3, TIMP-4, Bcl-2, Bax and Bcl-xL of CRC. In addition, terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick end-labeling (TUNEL) and hematoxylin and eosin staining were also conducted to examine cell apoptosis and necrosis. It was found that grafted human colorectal tumor growth and metastasis were obviously inhibited while tumor cell apoptosis and necrosis were induced after in vivo APRIL siRNA injection into nude mice. The data indicated that silencing of the APRIL gene using RNAi may serve as a novel therapeutic strategy for treatment of CRC.

Characteristics of hepatic igf-ii expression and monitored levels of circulating igf-ii mRNA in metastasis of hepatocellular carcinoma.

The prognosis of hepatocellular carcinoma (HCC) remains dismal. Insulin-like growth factor II (IGF-II), a fetal growth factor, is highly expressed during HCC development. We examined serum IGF-II levels and circulating IGF-II messenger RNA (mRNA) expression and analyzed the clinicopathologic characteristics in patients with liver diseases. The higher IGF-II level in the serum of patients with HCC could be correlated with hepatitis B virus infection but not with patient sex, age, tumor size, or α-fetoprotein (AFP) level. Total RNAs were extracted from liver tissues or peripheral blood mononuclear cells, and IGF-II complementary DNA (cDNA) and AFP cDNA were synthesized through random primers and reverse transcriptase; gene fragments were amplified by nested polymerase chain reaction and confirmed by sequencing. The incidence of the hepatic IGF-II gene was 100% in HCC, 54.3% in paracancerous tissues, and none in noncancerous tissues. The incidence rates for circulating IGF-II and AFP genes were 34.3% and 52.7%, respectively, and for both, 61.6% in patients with HCC. They were 100% in cases with extrahepatic metastasis. The IGF-II abnormality associates with HCC, and circulating IGF-II and IGF-II mRNA are useful molecular markers for HCC differential diagnosis and hematogenous metastasis.

A proliferation-inducing ligand: a new biomarker for non-small cell lung cancer.

To identify a proliferation-inducing ligand (APRIL) expression profile in tumor tissue and sputum of lung cancer, and evaluate the possibility of an assistant diagnosis of lung cancer by real-time fluorescence quantitative polymerase chain reaction (RTQ-PCR) in sputum as well, the authors analyzed the expression of APRIL mRNA in 75 tissue samples and 71 corresponding sputum samples of lung cancer by RTQ-PCR and analyzed their correlation. APRIL protein expression was also observed in tumor tissues by Western blot and immunohistochemistry. The expression analysis revealed APRIL expression was elevated in non-small cell lung cancer (NSCLC) and the expression of APRIL protein was located in the membrane and cytoplasm of tumor cells by immunohistochemiscal staining. Compared to benign pulmonary disease and healthy volunteers, the expression of APRIL mRNA in sputum of lung cancer was elevated (both P <. 001). When cut-off values for positivity were set at the mean + 2SD of mRNA expression in healthy volunteers, the positive rate for APRIL mRNA expression was 81.7% (58/71) in sputum samples of lung cancer, 3.2% (2/62) in benign pulmonary disease, and 1.5% (1/65) in healthy volunteers. The correlation was evident between the expression level of APRIL mRNA of tissue samples and that of sputum samples (P <. 001, r =. 702). These results support the possibility that the APRIL gene may play a key role in lung cancer, especially in NSCLC. The elevated expression level of APRIL mRNA in sputum of NSCLC suggested that APRIL mRNA may serve as an effective and convenient diagnostic biomarker for NSCLC.

[Expression of nuclear export factor CRM1 and p27 in glioma].

OBJECTIVE: To investigate the expression of nuclear export factor CRM1, Ser10-phosphorylated p27 and p27 in human gliomas. METHODS: The expression of CRM1, Ser10-phosphorylated p27 and p27 were investigated in 70 cases of human gliomas and 10 specimens of the normal brain tissue by immunohistochemical technique and Western blot. RESULTS: There were significant differences on the expression levels of CRM1, Ser10-phosphorylated p27 and p27 among normal brain tissue, gliomas of grades II and gliomas of grades III plus IV (P < 0.01). The expression of CRM1 in gliomas was inversely correlated with the expression of p27 (r(s) = -0.727, P < 0.01) and positively correlated with the expression of Ser10-phosphorylated p27 (r(s) = 0.954, P < 0.01) and Ki-67 (r(s) = 0.799, P < 0.01). Moreover, the expression of Ser10-phosphorylated p27 was inversely correlated with p27 (r(s) = -0.744, P < 0.01) and positively correlated with Ki-67 (r(s) = 0.785, P < 0.01). CONCLUSIONS: CRM1, through recognizing and binding with Ser10-phosphorylated p27, may promote moving of p27CRM1 from its original locating sites; act as a critical signaling component in the proliferative process of glioma cells and then, plays an important role in the development of gliomas.

[Correlation of hTERT expression to maspin and bFGF expression and their significance in glioma].

BACKGROUND & OBJECTIVE: Human telomerase reverse transcriptase (hTERT) is a determinant factor in telomerase activation and is related with tumorigenesis and the degree of malignancy. Its expression is regulated by many factors. This study was to detect the expression of hTERT, maspin, and basic fibroblast growth factor (bFGF) in glioma, and analyze their correlations and their significance in glioma. METHODS: The expression of hTERT, maspin, and bFGF in 128 specimens of human glioma and 8 specimens of normal brain tissue were detected by in situ hybridization and SP immunohistochemistry. H-score system of Gatalica was used for semi-quantitative evaluation. The positive rates between the 2 groups was compared with Chi(2) test. The correlations of hTERT, maspin, and bFGF expression to tumor grade were analyzed by Spearman rank correlation analysis. The correlations of hTERT expression to maspin and bFGF expression were analyzed by linear correlation. RESULTS: The positive rates of hTERT, maspin, bFGF were 51.6%, 46.9%, and 62.5% respectively in gliomas, and 0, 87.5%, and 0 in normal brain tissues. In the 43 specimens of grade II, 55 specimens of grade III and 30 specimens of grade IV gliomas, the positive rates of hTERT were 32.6%, 54.5%, and 73.3% (P < 0.05); the positive rates of maspin were 58.1%, 49.1%, and 26.7% (P < 0.05); the positive rates of bFGF were 39.5%, 72.7%, and 76.7% (P < 0.05).The expression of hTERT and bFGF were positively correlated to pathologic grade (rho=0.515, P < 0.01; rho=0.611, P < 0.01), while the expression of maspin was negatively correlated to pathologic grade (rho=-0.425, P < 0.05). hTERT expression was negatively correlated to maspin expression (r=-0.658, P<0.01), but positively correlated to bFGF expression (r=0.627, P < 0.01); maspin expression was negatively correlated to bFGF expression (r=-0.501, P < 0.01). The expression of hTERT showed no relationship with the age, sex, tumor size, and cell density (P > 0.05), but had obvious relationship with karyokinesis, vessel density, and necrosis (P < 0.05). CONCLUSION: The expression of hTERT, maspin and bFGF correlate to each other, and associate with the malignant degree of glioma.

Quantitative analysis of vascular endothelial growth factor, microvascular density and their clinicopathologic features in human hepatocellular carcinoma.

BACKGROUND: Angiogenesis is known to be essential to the survival, growth, invasion, and metastasis of tumor cells. Vascular endothelial growth factor (VEGF) are an important angiogenic factor regulating tumor angiogenesis, but its significance and tumor pathologic features are unclear in hepatocellular carcinoma (HCC). In the present study, we analyzed expression of tissue VEGF, alteration of microvascular density (MVD) in microvessel angiogenesis, development and metastasis of HCC, and level of serum VEGF in differential diagnosis of benign and malignant liver diseases. METHODS: Tumor specimens were prospectively collected from HCC patients undergoing resection. Total RNAs were extracted and the expression levels were detected from different parts of HCC tissues. The cellular distributions of VEGF and MVD of liver tumors and their paracancerous and distal cancerous tissues were investigated by streptavidin peroxidase (S-P) immunohistochemistry, respectively. The VEGF levels of circulating blood and hepatoma tissues were measured by enzyme-linked immunosorbent assay. RESULTS: The incidence of VEGF expression was 63.9% in HCCs (23/36 cases), 78.3% in non-encapsulated HCCs (18/23), and 90.9% in HCCs with extrahepatic metastasis (10/11), respectively. The VEGF expression was tightly correlated with MVD (P<0.01). The MVD in HCC with metastasis, low differentiation or non-encapsulation was significantly higher than that in HCC with intact capsule, high differentiation, or no metastasis. No significant difference was found between VEGF, MVD, tumor size, and hepatitis virus infection. The level of total RNA in HCC tissues was significantly lower but the VEGF level significantly higher than those in paracancerous or distal cancerous ones (P<0.01). The abnormal expression levels of VEGF in sera of HCC patients were directly correlated with the metastasis and recurrence of tumors. CONCLUSION: The high expression of VEGF and abnormality of tissue MVD are useful predictors for vascular invasion and metastasis of liver tumors.