The complete mitochondrial genome of two different colour Luciobarbus capito (Cypriniformes, Cyprinidae).

We sequenced and characterized the complete mitochondrial genome from normal colour (grey black) and mutant colour (orangey red) of Luciobarbus capito. Both mitogenomes contained the typical complement of 13 protein-coding genes, 22 transfer RNAs (tRNAs), two ribosomal RNAs (rRNAs), and a non-coding control region. They share the same gene arrangement pattern that was identical with most vertebrates. The entire mitochondrial DNA molecule of grey black L. capito was 16603-bp long, while the complete mtDNA molecule of orangey red L. capito was 16607-bp long.

The complete mitochondrial genome of golden yellow snakehead fish, Channa argus.

We sequenced and characterized the complete mitochondrial genome of golden yellow snakehead fish, Channa argus. The mitogenomes contained the typical complement of 13 protein-coding genes, 22 transfer RNAs (tRNAs), 2 ribosomal RNAs (rRNAs), and a non-coding control region. They share the same gene arrangement pattern that was identical with most vertebrates. The entire mitochondrial DNA molecule of golden yellow snakehead fish was 16,558 bp long. All information reported in this article will be a useful source of sequence information for general molecular and evolutionary studies of the family Channidae.

Musashi-2 promotes hepatitis Bvirus related hepatocellular carcinoma progression via the Wnt/ß-catenin pathway.

Our recent study observed that the expression of Musashi-2 (MSI2), a member of the Musashi family, was up-regulated in hepatitis B virus (HBV) related hepatocellular carcinoma parenchymal cells. Using quantitative PCR, tissue microarray (TMA) and immunohistochemical staining, we evaluated MSI2 mRNA and protein levels in tumor tissues from patients with different stages of hepatocellular carcinoma with paired adjacent noncancerous sample sets. The following techniques were used to further investigate MSI2 function and its potential molecular mechanism: RNAi, wound healing assay, Transwell assay, quantitative PCR and western blot analysis. Immunohistochemical detection of MSI2 on a TMA containing 106 paired specimens showed that increased cytoplasmic and nuclear MSI2 staining was significantly associated with tumor size, tumor differentiation, recurrence, TNM stage, vessel invasion and Ki-67 proliferative index. Patients with MSI2-positive tumors had a significantly higher disease recurrence rate and poorer survival than patients with MSI2-negative tumors after radical surgery. Based on univariate analysis, MSI2 expression showed an unfavorable influence on both disease-free survival and overall survival. Multivariate analysis revealed that higher MSI2 expression, together with tumor size, tumor differentiation, tumor thrombus, and Ki-67 expression were independent predictors of overall survival. With MSI2 knockdown, hepatoma cell migration and invasion were inhibited and the expression of ß-catenin, T cell factor (TCF) and lymphoid enhancer factor (LEF) were dysregulated. Thus, we propose that MSI2 may predict unfavorable outcomes in hepatitis B virus related hepatocellular carcinoma and promote cancer progression via the Wnt/ß-catenin signaling pathway.

Prognostic significance of SERPINE2 in gastric cancer and its biological function in SGC7901 cells.

PURPOSE: Altered expression of serine protease inhibitor peptidase inhibitor clade E member 2 (SERPINE2) associates with human cancer development and progression; thus, this study investigated SERPINE2 expression in gastric cancer tissues for association with clinicopathological and survival data from the patients and then investigated the role of SERPINE2 in gastric cancer cells in vitro. METHODS: The levels of SERPINE2 mRNA in 243 gastric cancer tissues and paired non-cancerous mucosa were determined using quantitative PCR. Inhibition of SERPINE2 expression by small interfering RNA (siRNA) was detected by Western blotting. tetrazolium, soft agar, and transwell assays were performed to evaluate the proliferation, anchorage-independent growth, and motility of gastric cancer SGC7901 cells transfected with SERPINE2 siRNA. RESULTS: Compared with the normal mucosa, SERPINE2 mRNA was increased in gastric cancer tissues and cells. Analysis of the 243 matched specimens showed that high SERPINE2 levels were associated with lymph node metastasis, distant metastasis, and clinical stage. Patients with high SERPINE2 mRNA levels had poorer survival compared with patients with low SERPINE2 mRNA levels. In vitro, SERPINE2 inhibited anchorage-independent growth, migration, and invasion of SGC7901 cells, but not proliferation. CONCLUSIONS: Our findings indicate that upregulated SERPINE2 may contribute to the aggressive phenotype of gastric cancer and suggest that SERPINE2 can be used as a novel prognostic factor and anticancer target in patients with gastric cancer.

Ciz1 is a novel predictor of survival in human colon cancer.

Cip1-interacting zinc-finger protein1 (Ciz1) is a nuclear matrix protein associated DNA replication factor which has been implicated in breast and lung cancer progression. However, the clinical significance of Ciz1 expression in colon cancer has not been determined. This study aimed to examine Ciz1 expression pattern and its potential as a biomarker of prognosis in colon cancer. Using quantitative PCR, tissue microarray (TMA), and ELISA, we evaluated Ciz1 mRNA and protein levels in tumor tissues from patients with colon cancer and in paired adjacent normal tissues. Ciz1 mRNA expression was significantly upregulated in 22 of 39 paired samples (P < 0.001). Immunohistochemistry on TMA-containing samples from 203 colon cancer patients indicated that Ciz1 protein expression was significantly higher in tumor tissues than in adjacent normal tissues (Stuart-Maxwell test, P < 0.001). Elevated expression of Ciz1 protein was significantly correlated with T stage (P < 0.001), N stage (P = 0.005), M stage (P = 0.021), and AJCC stage (P = 0.002). Multivariate Cox proportion hazard model analysis revealed that Ciz1 expression is an independent prognostic factor for overall time (OS; hazard ratio (HR): 1.76; 95% confidence interval (CI): 1.04-2.98; P = 0.034) and disease-free survival (DFS; HR: 2.02; 95% CI: 1.14-3.58; P = 0.017) of patients with colon cancer after colectomy. Our data suggested that Ciz1 may be involved in colon cancer progression and could serve as a novel predictor of survival for colon cancer patients.

Construction of leaky strains and extracellular production of exogenous proteins in recombinant Escherichia coli.

In this study, a strategy of the construction of leaky strains for the extracellular production of target proteins was exploited, in which the genes mrcA, mrcB, pal and lpp (as a control) from Escherichia coli were knocked out by using single- and/or double-gene deletion methods. Then the recombinant strains for the expression of exogenous target proteins including Trx-hPTH (human parathyroid hormone 1-84 coupled with thioredoxin as a fusion partner) and reteplase were reconstructed to test the secretory efficiency of the leaky strains. Finally, the fermentation experiments of the target proteins from these recombinant leaky strains were carried out in basic media (Modified R media) and complex media (Terrific Broth media) in flasks or fermenters. The results demonstrated that the resultant leaky strains were genetically stable and had a similar growth profile in the complex media as compared with the original strain, and the secretory levels of target proteins into Modified R media from the strains with double-gene deletion (up to 88.9%/mrcA lpp-pth) are higher than the excretory levels from the strains with single-gene deletion (up to 71.1%/lpp-pth) and the host E. coli JM109 (DE3) (near zero). The highest level of extracellular production of Trx-hPTH in fermenters is up to 680 mg l(-1).

PPAR-γ and tollip are associated with toll-like receptors in colitis rats.

To elucidate the significance of Toll-like receptors and their negative regulating factors PPAR-γ and Tollip on the pathogenesis of colitis. Colitis model was induced by TNBS in rat. The expression of TLR2, TLR4, NF-κBp65, PPAR-γ and Tollip was examined by immunohistochemistry (IHC) and reverse-transcription polymerase chain reaction (RT-PCR). RT-PCR revealed a significant increased expression of TLR2, TLR4, and NF-κBp65 in the colitis group compared with the normal group (TLR2: 1.057 ± 0.092, 0.463 ± 0.101, t = 4.125, P = 0.001; TLR4: 0.376 ± 0.029, 0.215 ± 0.049, t = 2.731, P = 0.013; NF-κBp65: 0.746 ± 0.049, 0.206 ± 0.063, t = 6.055, P = 0.000). The expression was positively correlated with the generally damage score and the histological injury score correspondingly (TLR2: r = 0.573, r = 0.559; TLR4: r = 0.754, r = 0.866; NF-κBp65: r = 0.548, r = 0.919). The Tollip mRNA wasn't obviously diversity between the normal and colitis groups by RT-PCR (Tollip: 0.288 ± 0.050, 0.140 ± 0.046, t = 1.993, P = 0.061). While the Tollip protein was mainly assembled in the lamina propriaand higher in the colitis group compared with the normal group by IHC. The expression of PPAR-γ in the colitis group was obviously lower than that in the normal group (PPAR-γ: 0.255 ± 0.065, 0.568 ± 0.072, t = 2.882, P = 0.010). The expression of Tollip and PPAR-γ was negative correlated with the generally damage score and histological injury score correspondingly (Tollip: r = -0.497, r = -0.551; PPAR-γ: r = -0.683, r = -0.853). The disbalance between TLRs and their negative regulating factors PPAR-γ and Tollip was closely associated with the course of colitis.

[Ecological niche breadth and niche overlap of dominant species of fish assemblage in Yangtze river estuary and its adjacent waters].

Based on the fishery resources data from the bottom trawl surveys conducted on the R/V Beidou in the Yangtze River estuary and its adjacent waters in June, August and October of 2006, the index of relative importance (IRI) was measured to determine the dominant species of fish assemblage, and the niche indicators and their seasonal variations of the dominant species were analyzed. A total of 10 dominant species in the 3 survey cruises were recorded, which were divided into two groups by the Bray-curtis similarity clustering and non-metric multidimensional scaling (MDS) analysis, with a significant seasonal variation of niche breadth and niche overlap. One group included Engraulis japonicus, Champsodon capensis, and Acropoma japonicum, whose niche breadth and niche overlap were larger in summer than in autumn, with a migration from the Yangtze River estuary and its adjacent waters to outer deeper waters, while the other group included Trichiurus haumela, Chaeturichthys stigmatias, Apogon lineatus, Larimichthys polyactis, Psenopsis anomala, Argyrosomus argentatus, and Benthosema pterotum, whose niche breadth and niche overlap were larger in autumn than in summer, with a reverse migration from southern Yellow Sea and northern East China Sea to the Yangtze River estuary and its adjacent waters. The different migration direction of the two groups was related to their ecological habits and environmental factors.

[Association between gut-enriched Kruppel-like factor and prognosis of patients with gastric cancer].

OBJECTIVE: To explore the expression of gut-enriched Kruppel-like factor 4(KLF4) in gastric cancer, and its association with prognosis. METHODS: Surgical specimens were collected from 264 patients undergoing radical surgery between 2004 and 2009 in the Affiliated Qianfoshan Hospital, Shandong University. KLF4 mRNA level of specimens was detected by real-time PCR. KLF4 protein expression was measured by immunohistochemistry on tissue microarray, which contained primary gastric cancer, corresponding para-cancerous tissue, and paired lymph node metastases. RESULTS: Real-time PCR revealed that mRNA level of KLF4 was down-regulated in gastric cancer compared with paired normal gastric mucosa. Immunohistochemistry on tissue microarray showed gastric cancer tissues had significantly lower KLF4 levels compared with paired normal gastric tissues. By univariate and multivariate analysis, KLF4 was a significant predictor of survival and recurrence. CONCLUSION: KLF4 expression is significantly down-regulated in gastric cancer, and is an independent predictor of survival and recurrence.

Downregulation of metallothionein 1F, a putative oncosuppressor, by loss of heterozygosity in colon cancer tissue.

PURPOSE: Downregulation of metallothionein (MT) genes has been reported in several tumors with discrepant results. This study is to investigate molecular mechanism of MT gene regulation in colon cancer which is characterized by tumor suppressor gene alterations. EXPERIMENTAL DESIGN: Integral analysis of microarray data with loss of heterozygosity (LOH) information was employed. Quantitative real-time PCR and immunohistochemistry were used to validate MT isoform expression in colon cancer tissues and cell lines. The effects of MT1F expression on RKO cell survival and tumorigenesis was analyzed. Bisulphite sequencing PCR (BSP) and methylation-specific PCR were employed to detect the methylation status of the MT1F gene in colon cancer tissues and cell lines. DNA sequencing was used to examine the LOH at the MT1F locus. RESULTS: MT1F, MT1G, MT1X, and MT2A gene expression was significantly downregulated in colon cancer tissue (p<0.05). Exogenous MT1F expression increased RKO cell apoptosis and inhibited RKO cell migration, invasion and adhesion as well as in vivo tumorigenicity. Downregulation of MT1F gene in majority of human colon tumor tissues is mainly through mechanism by loss of heterozygosity (p=0.001) while CpG island methylation of MT1F gene promoter region was only observed in poorly differentiated, MSI-positive RKO and LoVo colon cancer cell lines. CONCLUSIONS: MT1F is a putative tumor suppressor gene in colon carcinogenesis that is downregulated mainly by LOH in colon cancer tissue. Further studies are required to elucidate a possible role for MT1F downregulation in colon cancer initiation and/or progression.

Biglycan expression correlates with aggressiveness and poor prognosis of gastric cancer.

Biglycan, a member of the small leucine-rich proteoglycan family, has been implicated in the development and progression of human cancers. However, the clinical significance of biglycan expression in gastric cancer has not been determined. In the present study, biglycan mRNA and protein concentrations were analyzed using quantitative realtime reverse transcription polymerase chain reaction and Western blot in 69 gastric cancer and adjacent non-tumorous tissues, respectively. Biglycan expression was further assessed using immunohistochemistry in tissue microarrays that contained 264 cases of gastric cancer, and others containing normal or metastasized lymph node tumor tissues. Biglycan was upregulated at the transcriptional and translational levels and there was a correlation between the expression of biglycan mRNA and protein (P = 0.000, κ = 0.769). Over-expression of biglycan was strongly associated with lymph node metastasis, tumor (T) classification, metastasis (M) classification, vascular invasion and Union for International Cancer Control (UICC) stage. Patients with biglycan-positive tumors had a significantly higher disease recurrence rate and poorer survival than patients with biglycan-negative tumors after the radical surgery. Multivariate analysis revealed that biglycan expression is an independent prognostic indicator for survival of patients with gastric cancer. The data from the current study demonstrate that elevated expression of biglycan may play an important role in the development and progression of gastric cancer, and could be further evaluated as a biomarker for predication of a poor clinical outcome.

Expression of Toll-like receptor 4, CD14, and NF-κB in Chinese patients with ulcerative colitis.

This study elucidates the significance of Toll-like receptor 4 (TLR4), CD14, and nuclear factor (NF)-κB on the pathogenesis of ulcerative colitis (UC). Colonic biopsy specimens were collected from active UC and controls. The expression of TLR4, CD14, and NF-κBp 65 was analyzed by immunohistochemistry (IHC) and reverse-transcription polymerase chain reaction (RT-PCR). In UC, disease activity index (DAI) and pathological grade were classified according to the Powell-Tuck grade system and Truelove-Richards system, respectively. Fifty-six UC cases and 56 controls entered the investigation. IHC and RT-PCR revealed a significant increase of TLR4, CD14, and NF-κBp 65 antigen expression in colonic mucosa of UC compared with colonic mucosa of controls (p < .001). In UC, TLR4, CD14, and NF-κBp 65 expression were positively related to DAI (r = .873, p < .001; r = .576, p < .001; r = .747, p < .001 receptively). NF-κBp65 significantly correlated with TLR4 and CD14 (r = .669, p < .001; r = .576, p < .001, receptively). TLR4, CD14, and NF-κBp65 were positively related to pathological classification in UC (p < .01). Thus, TLR4, CD14, and NF-κBp65 were upregulated significantly in UC, to an extent that reflects the degree of inflammation and thereby might contribute to the occurrence and development of UC.

[The expression of toll-like receptor4 and NF-kappaB in ulcerative colitis].

AIM: To investigate the expression of TLR4 and NF-kappaB in ulcerative colitis (UC). METHODS: Colonic biopsy specimens were collected from active UC and controls. The expression of TLR4 and NF-kappaBp 65 were analyzed by immunohistochemistry (IHC) and RT-PCR. RESULTS: RT-PCR revealed a significant increase of TLR4 and NF-kappaBp 65 antigen expression in colonic mucosa of UC compared with colonic mucosa of controls (TLR4: 143.658+/-33.870, 30.531+/-8.442, t=24.253, P<0.01; NF-kappaBp65: 185.773+/-37.625, 23.810+/-7.038, t=31.664, P<0.01). IHC show that the expression of TLR4 and NF-kappaB was significantly higher in colonic mucosa of UC compared with colonic mucosa of controls. CONCLUSION: The expression of the TLR4 and NF-kappaB was increased in the colonic mucosa of UC compared with colonic mucosa of controls. It may be closely associated with the course of ulcerative colitis.

[The relation of peroxisome proliferator activated receptorbeta and Epithelial Ovarian Carcinoma].

AIM: To explore the possible relationship between expression of PPAR(pevoxisome prodiferator adivated receptor) beta and clinical pathological features of epithelial ovarian carcinoma (EOC). METHODS: PPARbeta expressions in normal ovaries (n=10), ovarian bordline tumors (n=10), and ovarian serous cystadenocarcinomas (n=46) were investigated by immunohistochemistry and RT-PCR. RESULTS: PPARbeta protein was expressed in normal ovaries and ovarian cancer. PPARbeta protein was found in cytoplasm of tumor cell. PPARbeta expression was 20% revealed in normal ovaries, 50% in bordline tumors 89.1% in serous cystadenocarcinomas(P<0.05), significantly higher than that in bordline tumors (P<0.05). PPARbeta expression was positively correlated with tumor stage, histological grade and lymphonode metastasis (P<0.05). CONCLUSION: High expression of PPARbeta may be related with the differentiation and metastasis of epithelial ovarian carcinoma.

[Differential expression of USP2, USP14 and UBE4A between ovarian serous cystadenocarcinoma and adjacent normal tissues].

AIM: The differential expression of USP2, USP14 and UBE4A between ovarian serous cystadenocarcinoma and adjacent normal tissues was investigated. METHODS: Restriction fragment differential display polymerase chain reaction (RFDD-PCR), semi-quantitative RT-PCR and immunohistochemical staining were applied to analyze the differentially expressed genes and proteins of ubiquitin specific proteases (USPs), USP2 and USP14, and ubiquitin factor E4A (UBE4A) between ovarian serous cystadenocarcinoma and adjacent normal tissues obtained from 40 patients aged from 29 to 72 years old, collected in 2005 year at excision of surgical operation with ovarian serous cystadenocarcinoma. RESULTS: USP2, USP14 and UBE4A were over-expressed (>3 folds) in ovarian serous cystadenocarcinoma tissues compared to normal tissues. CONCLUSION: The results suggest that the activity of ubiquitin-proteasome system is obviously enhanced in ovarian cancer.

Role of beta-catenin in hepatocarcinogenesis of rats.

BACKGROUND: Beta-catenin has two distinct roles in E-cadherin mediated cell adhesion and carcinogenesis by activating the wnt/beta-catenin signaling pathway. One occurs at the cell-adhesion site, where cadherins are linked to the actin-based cytoskeleton. The other takes place in the cytoplasm and nuclei and is thought to regulate cell transformation. We studied the role of beta-catenin in hepatocarcinogenesis of rats. METHODS: Fresh liver specimens were obtained from normal rats, and atypical hyperplasia livers and hepatocarcinoma tissues from model rats. The changes of beta-catenin in gene expression levels were detected by reverse transcriptase polymerase chain reaction (RT-PCR) in the different specimens separately. At the same time, their localization was observed immunohistochemically. RESULTS: In the normal liver specimens, beta-catenin staining was seen in the cell membrane. In liver specimens of atypical hyperplasia, beta-catenin staining occurred in the cell cytoplasm of some cells as well as in the cell membrane of others. Immunohistochemically cancerous tissues showed the presence of beta-catenin in the cytoplasm and nuclei. RT-PCR revealed that the gene expression levels of beta-catenin were same in all samples. CONCLUSIONS: The accumulation of beta-catenin in the cytoplasm and/or nuclei frequently occurs in hepatocarcinogenesis of rats. It may be an early event in the development of hepatocarcinoma of rats.

Combined devascularization and proximal splenorenal shunt: is this a better option than either procedure alone?

BACKGROUND/PURPOSE: We aimed to determine the rationality of pericardial devascularization (PCDV) plus proximal splenorenal shunt (PSRS) for cirrhotic patients with portal hypertension with variceal bleeding, using a duplex sonography study of the effects of the different surgical procedures (PCDV, PSRS, and PCDV + PSRS) on the hemodynamics of the portal system. METHODS: Ninety-nine patients with cirrhotic portal hypertension and a history of bleeding esophageal varices were studied. These patients were divided into three groups (PSRS group, PCDV group, and PCDV + PSRS group). The hemodynamic parameters of the portal systems of all patients were measured by Doppler color-flow imaging perioperatively. RESULTS: In the PSRS group, the postoperative portal venous flow (PVF) and free portal pressure (FPP) decreased by 57 +/- 9% and 52 +/- 5%, respectively (P < 0.01). In the PCDV group, the postoperative PVF lessened by 8 +/- 5% (P > 0.05), and the postoperative FPP was reduced by 19 +/- 7% (P < 0.05). In the PCDV + PSRS group, the postoperative PVF and FPP were lowered by 36 +/- 8% and 34 +/- 10%, respectively (P < 0.05). The postoperative decreases of PVF and FPP in the PCDV + PSRS group were between those of the PSRS and PCDV groups. The differences among these groups were statistically significant (P < 0.05). CONCLUSIONS: Combined devascularization and splenorenal shunt (PCDV + PSRS) significantly decreases portal venous flow and portal pressure, as well as maintaining hepatopedal flow, thus entailing fewer complications compared to either PCDV or PSRS. We aimed to determine the rationality of pericardial devascularization (PCDV) plus proximal splenorenal shunt (PSRS) for cirrhotic patients with portal hypertension with variceal bleeding, using a duplex sonography study of the effects of the different surgical procedures (PCDV, PSRS, and PCDV + PSRS) on the hemodynamics of the portal system.