miR-125b regulates side population in breast cancer and confers a chemoresistant phenotype.

Resistance to chemotherapy is a major obstacle for the effective treatment of breast cancer and is partially due to the presence of drug resistant stem cell-like side population (SP). Previous studies have shown elevated miR-125b is associated with chemoresistance and metastasis; however, the relationship between miR-125b and SP cells remains unknown. In this study, we isolated and characterized SP cells in a panel of breast cancer cell lines and primary cancer cells from breast cancer patients. SP cells showed cancer stem cells (CSCs) properties, including self-renewal, resistance to chemotherapy and high expression of stem cell markers. The percentage of SP cells was higher in chemotherapy resistant patients compared to that in chemotherapy responsive patients (5.8 ± 2.4% in non-responsive patients vs. 1.2 ± 0.5% in responsive patients, P = 0.012). Importantly, SP cells had higher level of miR-125b than NSP cells and the elevated miR-125b expression in chemoresistant cancer cells were due to high percentage of SP cells. Overexpression of miR-125b correlated with an increase in tumor SP and CSC property, whereas knockdown of miR-125b correlated with decreased incidence of SP. In addition, miR-125b overexpression in breast cancer cells induced epithelial-mesenchymal transition (EMT)-like cellular marker alteration, suggesting a potential mechanism of miR-125b in the regulation of cancer stem-like SP cells. Taken together, these results suggest an important role for miR-125b in breast cancer chemoresistance by maintaining cancer stem-like SP fraction, and raise the possibility that miR-125b may be a significant prognostic response marker for cancer therapy.

Non-traumatic diaphragmatic hernia of the liver in an adult: a case report.

BACKGROUND: Diaphragmatic hernia of the liver is a rare clinical entity, usually found after trauma in adults. This study was undertaken to elucidate a misdiagnosis of non-traumatic diaphragmatic hernia of the liver in an adult. METHOD: The clinical data of one patient with non-traumatic diaphragmatic hernia of the liver was analyzed. RESULTS: A tumor in the right lower thorax was revealed by chest X-ray and computed tomography. Non-traumatic diaphragmatic hernia of the liver was not identified until the operation. Pathological analysis confirmed the finding. The patient recovered well. CONCLUSIONS: Non-traumatic diaphragmatic hernia of the liver in an adult is a rare right-sided diaphragmatic hernia, which can move up into the chest cavity. It should be distinguished from lung cancer. The diagnosis and evaluation of non-traumatic diaphragmatic hernia of the liver can help optimize surgical management.

Osthole inhibits the tumorigenesis of hepatocellular carcinoma cells.

Hepatocellular carcinoma (HCC) accounts for approximately 90% of all cases of primary liver cancer, and the majority of patients with HCC are deprived of effective curative methods. Osthole is a Chinese herbal medicine which has been reported to possess various pharmacological functions, including hepatocellular protection. In the present study, we investigated the anticancer activity of osthole using HCC cell lines. We found that osthole inhibited HCC cell proliferation, induced cell cycle arrest, triggered DNA damage and suppressed migration in HCC cell lines. Furthermore, we demonstrated that osthole not only contributed to cell cycle G2/M phase arrest via downregulation of Cdc2 and cyclin B1 levels, but also induced DNA damage via an increase in ERCC1 expression. In addition, osthole inhibited the migration of HCC cell lines by significantly downregulating MMP-2 and MMP-9 levels. Finally, we demonstrated that osthole inhibited epithelial-mesenchymal transition (EMT) via increasing the expression of epithelial biomarkers E-cadherin and ß-catenin, and significantly decreasing mesenchymal N-cadherin and vimentin protein expression. These results suggest that osthole may have potential chemotherapeutic activity against HCC.

Malignant nonfunctioning islet cell tumor of the pancreas with intrasplenic growth: a case report.

BACKGROUND: We reported a case of malignant nonfunction islet cell tumor (10.0 cm in diameter) of the pancreas, with malignant histological features and splenic infiltration. The case is rare, and few reports have been published. METHODS: A 46-year-old woman with a vague pain in the left upper quadrant for 3 months was found to have a tumor in the spleen. Ultrasonography and computed tomography demonstrated a well-defined pancreatic tumor of 8.2 X 10.0 cm in size, her serum levels of pancreatic hormones were within normal limits. RESULTS: Splenectomy combined with pancreatectomy was performed for the tail of the pancreas. Resected specimens showed a malignant nonfunctioning islet cell tumor invading the spleen. CONCLUSIONS: The growth pattern of the tumor causes malignant features. Resection of the tumor should be performed by enucleation, pancreaticoduodenectomy or distal pancreatectomy.

Diagnosis and management of extrahepatic portal vein aneurysm: a case report.

BACKGROUND: Extrahepatic portal vein aneurysm is a rare clinical entity with no more than 50 reported cases in the English literature. This study was to elucidate the procedures used in the diagnosis and management of portal vein aneurysm. METHOD: The clinical data of a patient with extrahepatic portal vein aneurysm were analyzed. RESULTS: Extrahepatic portal vein aneurysm associated with portal hypertension and splenomegaly was diagnosed by color Doppler ultrasonography (US), computed tomography (CT), magnetic resonance angiography (MRA) and angiography (indirect portography). Splenectomy and surgical shunt (splenorenal shunt) were performed. No hepatic encephalopathy and variceal bleeding occurred after the operation except mild pancreatic leakage caused by partial pancreatic tail resection. The patient recovered well. CONCLUSIONS: Extrahepatic portal vein aneurysm can be evaluated by US, CT, MRA, and angiography for optimum conservative or surgical management.

Gene expression changes after hypoxic preconditioning in rat hepatocytes.

BACKGROUND: Hypoxic preconditioning can protect hepatocytes against hypoxic injury, but its mechanism has not been elucidated. The aim of this study was to profile gene expression patterns involved in hypoxic preconditioning and probable mechanism at the level of gene expression. METHODS: Hepatocytes were divided into 2 groups: control group and hypoxic preconditioning group. Biotin-labeled cRNA from the control group and the hypoxic preconditioning group was hybridized by oligonucleotide microarray. Genes that were significantly associated with hypoxic preconditioning were filtered, and validated at the level of transcript expression. RESULTS: Forty-three genes with significantly altered expression patterns were discovered and most of them had not been previously reported. Among these genes, genes encoding superoxide dismutase 2 (SOD2) and interleukin 10 (IL-10) in the hypoxic preconditioning group were confirmed to be up-regulated with real-time quantitative PCR. CONCLUSIONS: Many cytokines are involved in hypoxic preconditioning and protect hepatocytes from hypoxia-reoxygenation injury, and the increase of oxygen free-radical scavengers and anti-inflammatory factors may play a key role in this phenomenon. Diverse signal pathways are probably involved.

Astragaloside IV Enhances Cisplatin Chemosensitivity in Human Colorectal Cancer via Regulating NOTCH3.

Although astragaloside IV exhibits anti-inflammation, immunoregulatory, and anticancer properties, the chemosensitization effects of astragaloside IV in colorectal cancer have never been reported. Our study tested whether astragaloside could increase cisplatin sensitivity in colorectal cancer. CCK-8 assay was used to measure the cell viability of colorectal cancer cells. Quantitative real-time PCR and Western blot were performed to determine the mRNA and protein expression, respectively. Our data revealed that astragaloside IV administration significantly suppressed the cell growth of colorectal cancer cells, whereas no obvious cytotoxicity of astragaloside IV was observed in nonmalignant colonic cells. In addition, combined treatment with astragaloside IV dramatically elevated the chemosensitivity of colorectal cancer cells to cisplatin. Mechanical investigation revealed that the mRNA and protein expression of NOTCH3 was significantly lower in cisplatin and astragaloside IV-treated cells compared with cells treated with cisplatin alone. On the contrary, no obvious changes in tumor cell growth were shown after upregulation of NOTCH3 whether in the presence or absence of astragaloside IV. Thus, our data demonstrate that astragaloside IV increases the chemosensitivity of colorectal cancer cells to cisplatin, at least partly, through inhibition of NOTCH3. This study suggests that combined therapy with astragaloside IV might be a novel therapeutic approach for colorectal cancer.

Changes in soil microbial community structure associated with two types of genetically engineered plants analyzing by PLFA.

With the rapid expansion of GEPs(genetically engineered plants), people are more and more concerned about the ecological risks brought by their release. Assessing the effect of GEPs on soil microbial ecology is indispensable to study their ecological risks. In our study, the phospholipids fatty acid ( PLFA) method was used to analyze the microbial community of soil samples collected from fields with two types of GEPs-Bt transgenic corn and PVY ( potato virus Y) cell protein gene transgenic potato. The principal components analysis (PCA) showed all controls were on the right of related GEPs samples along the PC1 (the first principal component) axis, which means a decrease of fungi in soils with genetically engineered crop since most of PLFAs that are strongly positively correlated with PC1 represent fungi. For samples collected from Bt transgenic cornfield, the ratios of gram-positive to gram-negative bacteria were less than those of controls. For samples of transgenic potato field, these ratios were lower than those of controls when soils were collected from deep layer (20-40 cm), but were higher when soils collected from surface layer(0-20 cm). For soils collected from 0-20 cm, the ratios of fungi to bacteria for all GEPs samples were at the same level. So were such rations for all controls. Changes of soil microbial community in two types of GEPs fields were detected in our study, but the causes and more information still needs further study.

Diversity surveys of soil bacterial community by cultivation--based methods and molecular fingerprinting techniques.

By combining the cultivation methods with molecular fingerprinting techniques, the diversity surveys of soil bacterial community in 13 areas of China were carried out. The cultivable heterotrophic diversity was investigated by colony morphology on solid LB medium. Genetic diversity was measured as bands on denaturing gradient gel electrophoresis(DGGE) by the extraction and purification of the total soil DNA, and amplification of bacterial 16S rDNA fragments by polymerase chain reaction (PCR). The Shannon-Wiener indices of diversity (H), richness (S) and evenness (E(H)) were employed to estimate the diversity of soil bacterial community. The results showed that there was an obvious diversification existed in soil from the different areas. However, the genetic diversity estimated by PCR-DGGE can provide more comprehensive information on bacterial community than the cultivation-based methods. Therefore, it is suggested to combine the traditional methods with genetic fingerprinting techniques to survey and estimate soil bacterial diversity.

[Experimental study on iNOS gene transfer mediated by liposome to treat portal hypertension in cirrhotic rats].

OBJECTIVE: To evaluate the effects of iNOS gene transfer on portal hypertensive rats. METHODS: Eukaryotic expression plasmid pcDNA(3)/iNOS was used to transfect sinusoidal endothelial cells (SEC) mediated by Lipofectamine. Transfection rate and gene exspression were detected. Hepatic cirrhosis was induced in male Sprague-Dawley rats by intraperitoneal injection of carbon tetrachloride, and the cirrhotic rats were divided into three groups:Liposome-pcDNA(3)/iNOS (n = 10), Tris buffer (n = 10) and nude plasmid (n = 10), which were injected into the portal vein of experiment cirrhotic rats respectively. Five days later, animals were killed, immunohistochemistry and spectrophotometry methods were used to measure the expression of iNOS and the amount of NO production. RESULTS: Eukaryotic expression plasmid pcDNA(3)/iNOS could effectively transfect SEC and express corresponding gene products. Following iNOS gene transfer, compared with the two controlled groups, iNOS expression and the NO production were significantly increased meanwhile portal pressure was decreased significantly. CONCLUSIONS: The iNOS gene transfer is a feasible and an effective approach to treat portal hypertension in cirrhotic rats which could increase the expression of intra-hepatic iNOS and the amount of NO production thus leading to a remarkable reduction of portal venous pressure.

Ubiquitin­specific protease 22­induced autophagy is correlated with poor prognosis of pancreatic cancer.

Ubiquitin­specific protease 22 (USP22) is a component of the transcription regulatory histone acetylation complex SAGA, which broadly regulates gene transcription and correlates with cancer progression, metastasis and prognosis. Autophagy is a cell pathway with dual functions that promotes cell survival or death. However, it is not known whether USP22 can regulate autophagy in pancreatic cancer. In the present study, we first identified that USP22 was overexpressed in a large number of pancreatic cancer patient samples, concomitant with the increased expression of LC3, a marker of autophagy. Statistical analysis revealed that the increase in USP22 and autophagy was positively correlated with poor prognosis of pancreatic cancer patients. Further investigation using a human pancreatic cancer cell (Panc­1) identified that the overexpression of USP22 increased the processing of LC3 into the active form LC3­II and the number of autophagosomes, thus leading to enhanced autophagy. Activation of ERK1/2 kinase rather than AKT1 by USP22 was found to be one of the mechanisms promoting LC3 processing. USP22­induced autophagy was also found to enhance cell proliferation and resistance to starvation and chemotherapeutic drugs in Panc­1 cells, therefore expressing an overall effect that promotes cell survival. Collectively, the present study demonstrated a new function of USP22 that induces autophagy, thus leading to the poor prognosis of pancreatic cancer.