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Sequence-related amplified polymorphism (SRAP) markers were used to assess the genetic relatedness among 33 Cynodon radiatus accessions from different regions of China. Fifteen primer combinations were used to amplify specific C. radiatus genomic sequences. A total of 382 SRAP fragments were generated, with fragment sizes ranging from 200 to 1800 bp. Genetic similarity coefficients among the 33 accessions ranged from 0.53 to 0.95, with an average of 0.70. Cluster analysis by two methods, the unweighted pair–group method with arithmetic averages and the principle coordinate analysis, separated the accessions into 3 distinct groups. This study shows that the SRAP technique is a reliable tool for differentiation of C. radiatus accessions and for determination of the genetic relationships among them.

Background & aims The discontinuation rules of transarterial chemoembolization (TACE) for patients who were assessed as progressive disease (PD) but stage progression-free (SP-free: still belongs to Barcelona Clinic Liver Cancer B) after TACE are unclear. We aimed to evaluate the impact of the PD-pattern on the survival of these patients retreated with TACE. Methods In total, 115 consecutive patients who were assessed as PD but SP-free after TACE and then underwent at least one subsequent TACE session were included. Sixty patients were assessed as PD with target lesion progression (TP), and 55 patients were assessed as PD with target lesion non-progression (TNP). Survival and treatment-related adverse events were compared between the two groups. Additional external validation was performed using a data set (n = 103) from another institution. Results Patients with TNP had significantly longer median post-progression survival (PPS) than those with TP (21.0 vs. 11.9 months, P = 0.004). After TACE retreatment, the incidence of liver dysfunction was significantly higher for patients with TP than for patients with TNP (45% vs. 20%, P = 0.031). In the multivariate analysis, the target lesion response was one of the most significant prognostic factors for PPS (HR = 2.01; 95% confidence interval: 1.23-3.27; P = 0.005). The findings were supported by an independent external cohort. Conclusions Compared to patients with TNP, patients with TP might exhibit no improvement in survival and even present damaged liver function after retreatment with TACE. Target lesion response is useful as a clinical decision for repeated TACE in these patients.

Background The optimal strategy for adjuvant therapy after curative resection for hepatocellular carcinoma (HCC) patients with solitary tumor and microvascular invasion (MVI) is controversial. This trial evaluated the efficacy and safety of adjuvant transcatheter arterial chemoembolization (TACE) after hepatectomy versus hepatectomy alone in HCC patients with a solitary tumor ≥ 5 cm and MVI. Methods In this randomized, open-labeled, phase III trial, HCC patients with a solitary tumor ≥ 5 cm and MVI were randomly assigned (1:1) to receive either 1–2 cycles of adjuvant TACE after hepatectomy (Hepatectomy-TACE) or hepatectomy alone (Hepatectomy Alone). The primary endpoint was disease-free survival (DFS); the secondary endpoints included overall survival (OS) and adverse events. Results Between June 1, 2009, and December 31, 2012, 250 patients were enrolled and randomly assigned to the Hepatectomy-TACE group (n = 125) or the Hepatectomy Alone group (n = 125). Clinicopathological characteristics were balanced between the two groups. The median follow-up time from randomization was 37.5 months [interquartile range 18.3–48.2 months]. The median DFS was significantly longer in the Hepatectomy-TACE group than in the Hepatectomy Alone group [17.45 months (95% confidence interval [CI] 11.99–29.14) vs. 9.27 months (95% CI 6.05–13.70), hazard ratio [HR] = 0.70 (95% CI 0.52–0.95), P = 0.020], respectively. The median OS was also significantly longer in the Hepatectomy-TACE group than in the Hepatectomy Alone group [44.29 months (95% CI 25.99–62.58) vs. 22.37 months (95% CI 10.84–33.91), HR = 0.68 (95% CI 0.48–0.97), P = 0.029]. Treatment-related adverse events were more frequently observed in the Hepatectomy-TACE group, although these were generally mild and manageable. The most common grade 3 or 4 adverse events in both groups were neutropenia and liver dysfunction. Conclusion Hepatectomy followed by adjuvant TACE is an appropriate option after radical resection in HCC patients with solitary tumor ≥ 5 cm and MVI, with acceptable toxicity.

Purpose To compare the stability of stable and unstable water-in-oil emulsions and the efficacy and safety of these emulsions in a single-center, prospective double-blind trial of transarterial chemoembolization for hepatocellular carcinoma (HCC). Materials and Methods A total of 812 patients with inoperable HCC were randomized (stable emulsion, n = 402; unstable emulsion, n = 410). The 2 emulsions were prepared by using the same protocol except that different solvents were used for chemotherapy agents, including epirubicin, lobaplatin, and mitomycin C. The solvent in the stable emulsion arm was contrast medium and distilled water, and the solvent in the unstable emulsion arm was distilled water. The primary endpoint was overall survival (OS), and secondary endpoints were time to progression (TTP), tumor response, adverse events (AEs), and plasma epirubicin concentrations. Results In vitro, stable emulsions did not occur until 1 day, and unstable emulsions, with a lower peak plasma concentration (P = .001) in vivo, exhibited rapid separation of the oil and aqueous phases after 10 minutes. Median OS times in the stable and unstable emulsion arms were 17.7 and 19.2 months, respectively (P = .81). No differences were found in TTP, tumor response, and AEs except for myelosuppression (anemia, 3.5% vs 7.6%; thrombocytopenia, 11.5% vs 17.7%), which was significantly more severe and frequent in the unstable emulsion arm (P = .013). Conclusions Chemoembolization is equally effective with the use of stable and unstable emulsions, but the use of a stable emulsion has the advantage of less myelosuppression and a favorable pharmacokinetic profile.

Adzuki bean (Vigna angularis) is an important legume crop native to China, but its rhizobia have not been well characterized. In the present study, a total of 60 rhizobial strains isolated from eight provinces of China were analyzed with amplified 16S rRNA gene RFLP, IGS-RFLP, and sequencing analyses of 16S rRNA, atpD, recA, and nodC genes. These strains were identified as genomic species within Rhizobium, Sinorhizobium, Mesorhizobium, Bradyrhizobium, and Ochrobactrum. The most abundant groups were Bradyrhizobium species and Sinorhizobium fredii. Diverse nodC genes were found in these strains, which were mainly co-evolved with the housekeeping genes, but a possible lateral transfer of nodC from Sinorhizobium to Rhizobium was found. Analyses of the genomic and symbiotic gene backgrounds showed that adzuki bean shared the same rhizobial gene pool with soybean (legume native to China) and the exotic Vigna species. All of these data demonstrated that nodule formation is the interaction of rhizobia, host plants, and environment characters.

T-2 toxin is one of the mycotoxins, a group of type A trichothecenes produced by several fungal genera including Fusarium species, which may lead to the decrease of testosterone secretion in primary Leydig cells derived from mouse testis. The previous study demonstrated T-2 toxin decrease the testosterone biosynthesis in the primary Leydig cells derived from the mouse testis directly. In this study, we further examined the direct biological effects of T-2 toxin on the process of steroidogenesis, primarily in Leydig cells of mice. Leydig cells of mature mouse were purified by Percoll gradient centrifugation and the cell purity was determined by 3β-hydroxysteroid dehydrogenase (3β-HSD) staining. To examine the decrease in T-2 toxin-induced testosterone secretion, we measured the transcription level of three key steroidogenic enzymes including 3β-HSD-1, cytochrome P450 side-chain cleavage (P450scc) enzyme, and steroidogenic acute regulatory (StAR) protein in T-2 toxin/human chorionic gonadotropin (hCG) co-treated cells. Our previous study showed that T-2 toxin (10−7, 10−8, and 10−9 M) significantly suppressed hCG (10 ng/ml)-induced testosterone secretion. The studies demonstrated that the suppressive effect is correlated with a decrease in the level of transcription of 3β-HSD-1, P450scc, and StAR ( p < 0.05).

// Jia-hong Wang 1, * , Wei Wei 1, * , Jing Xu 2 , Zhi-xing Guo 3 , Cheng-zuo Xiao 1 , Yong-fa Zhang 1 , Pei-en Jian 1 , Xiao-liang Wu 2 , Ming Shi 1 , Rong-ping Guo 1 1 Department of Hepatobilliary Oncology, Sun Yat-sen University Cancer Center, State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Guangzhou, China 2 State Key Laboratory of Oncology in South China, Sun Yat-sen University Cancer Center, Collaborative Innovation Center for Cancer Medicine, Guangzhou, China 3 Department of Ultrasonics, Sun Yat-sen University Cancer Center, State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Guangzhou, China * These authors have contributed equally to this work Correspondence to: Rong-Ping Guo, e-mail: guorp@sysucc.org.cn Keywords: hepatocellular carcinoma, Cripto-1, MMP-9, aggressiveness, prognosis Received: April 20, 2015 Accepted: August 24, 2015 Published: September 05, 2015 ABSTRACT Cripto-1 could promote tumorigenesis in a wide range of carcinomas, yet little is known in hepatocellular carcinoma (HCC). The expression of Cripto-1 and MMP-9 were assessed by immunohistochemistry in 205 HCC specimens. The correlation between Cripto-1 and MMP-9, clinicopathological/prognostic value in HCC was examined. Cripto-1 overexpression was correlated with larger tumor, TNM stage, BCLC stage and tumor recurrence. In multivariate analyses, Cripto-1 was an independent predictor for overall survival (OS) and time to recurrence (TTR). Cripto-1 expression was increased in TNM and BCLC stage-dependent manner. Cripto-1 overexpression was associated with poor prognosis in patients subgroups stratified by tumor size, tumor differentiation, TNM and BCLC stage. In addition, Cripto-1 was positively correlated with MMP-9 among 205 HCC samples. Patients with Cripto-1 upregulation had poor OS and shorter TTR in low and high aggressiveness groups. Furthermore, Cripto-1 had predictive validity for early and late recurrence in HCC patients. Combination of Cripto-1 and serum AFP was correlated with OS and TTR. In conclusion, Cripto-1 overexpression contributes to aggressiveness and poor prognosis of HCC. Cripto-1/AFP expression could be a potential prognostic biomarker for survival in HCC patients.

We have previously reported that Cezanne could be a prognostic biomarker for survival in hepatocellular carcinoma (HCC) patients. However, the role of Cezanne genes in HCC cells and its response to postoperative adjuvant transcatheter arterial chemoembolization (TACE) in HCC patients remains unknown. In this study, Cezanne expression was detected in human HCC using real-time PCR, western blot and immunohistochemistry. The function of Cezanne in HCC cells was determined by Transwell invasion assays and nude mice metastasis assay. The response of Cezanne in patients who received adjuvant TACE after hepatectomy was evaluated. Functional study demonstrated that interference of Cezanne expression promoted the migration and invasion of HCC cells in vitro and boosted metastasized HCC formation in mice. Upregulation of Cezanne diminished the adhesion and migration of hepatoma cells. Further study indicated that Cezanne might inhibit invasion of HCC cells by inducing epithelial–mesenchymal transition (EMT). In addition, patients with low Cezanne expression had significant improvement in prognosis after receiving adjuvant TACE. In contrast, patients with high Cezanne expression had a poorer response to adjuvant TACE. Moreover, Cezanne status was associated with response to adjuvant TACE in patients subgroup stratified by vascular invasion, tumor size and tumor number. In conclusion, Cezanne may be a novel antioncogene that has a pivotal role in the invasion of HCC and contribute to the selection of patients who may benefit from adjuvant TACE to prevent recurrence.

Background Transarterial chemoembolisation (TACE) is still widely used to treat patients with advanced hepatocellular carcinoma who have no access to sorafenib in Asia. However, the indication of TACE for these patients has yet to be established. We established an objective point score to guide this decision in patients with advanced-stage hepatocellular carcinoma. Methods We developed a scoring system (MCCT score) based on preoperative data from a cohort of 422 patients receiving TACE as the first treatment for advanced hepatocellular carcinoma from 2006 to 2013 at Sun Yat-sen University Cancer Center, Guangzhou, China. Factors determining survival were analysed by univariate and multivariate analysis and Cox proportional hazard regression models. The scoring system was retrospectively validated in an external cohort from other two institutions (Dongguan People's Hospital and the Kaiping Central Hospital; n=237). The prognostic predictive power was compared with six different staging systems: tumour node metastasis (TNM), 7th edition; Cancer of the Liver Italian Program (CLIP); Chinese University Prognostic Index (CUPI); Okuda, Japan Integrated Staging (JIS); and Groupe d'Etude et de Traitement du Carcinome Hepatocellulaire (GRETCH). The retrospective study was approved by the institutional review board; written informed consent was obtained from each patient. Findings The presence of macroscopic vascular invasion (M), Child-Pugh class B (C), C-reactive protein values (C) higher than 10 mg/L, and tumour size (T) larger than 7 cm were independent negative prognostic factors for survival and were used to create the MCCT score. The patients were divided into risk groups on the basis of their MCCT scores (low risk for ≤2 points, intermediate risk for 3–4 points, and high risk for five points). The median overall survival times were 14·4 months (95% CI 10·7–18·1) for the low-risk group, 6·8 months (6·0–7·6) of the intermediate-risk group, and 3·2 months (1·7–4·7) for the high-risk group (p Interpretation The MCCT scoring system is more accurate than the six other scoring systems in selecting patients with advanced hepatocellular carcinoma for TACE. For patients with advanced hepatocellular carcinoma with no access to sorafenib, TACE may be a valuable alternative in selecting patients (MCCT score ≤2). Funding This study was supported by grants from the National Natural Science Foundation of China (81172037/H1606, 81272639, and 81572385), Guangdong province science and technology project of China (2013B021800159), and the Fundamental Research Funds for the Central Universities of China (13ykzd24).

Basic data of four fractions, PM1, PM6, PM7and PM8, derived from Pingshuo bituminous coal were obtained by ultimate analyses, mean molecular weight measure, GC/MS, 13C-NM R, 1H-NM R and FTIR technigues. A method of calculating the average molecular parameters of the fractions is introduced. Based on the calculated parameters, the molecular structure models of the fractions of PM6, PM;and PM8are constructed. Significant structural information about the saturated hydrocarbon and aromatic hydrocarbon with heteroatom groups is given also.

Opioid receptors, members of the G-protein-coupled receptor superfamily, appear to be involved in cancer progression. However, the expression and significance of opioid receptors in esophageal squamous cell carcinoma (ESCC) remain unclear. In this study, we demonstrated by flow cytometry that μ, δ, and κ-opioid receptors (MOR, DOR, and KOR) are expressed to various degrees in ESCC cell lines. The KOR protein was further examined by several methods in ESCC cell lines and tissues. Immunocytochemical staining localized KOR to the cell membrane in KYSE180 cells and the nucleus in EC109 cells, whereas no signal or weak staining of the cytoplasm was observed in KYSE150 cells. The expression of KOR was confirmed in ESCC cells by Western blotting. Furthermore, immunohistochemistry staining showed that KOR was up-regulated in ESCC tissues compared with nontumorous esophageal epithelium (P = .004, χ(2) test). Moreover, high nuclear KOR expression was significantly correlated with lymph node metastasis in 256 ESCC cases (R = 0.144; P = .030, Kendall τB test). Patients with high nuclear KOR expression in ESCC had a significantly poorer prognosis (P = .001, log-rank test). Multivariate Cox analysis revealed that KOR in the nucleus was an independent prognostic factor (hazard ratio, 1.789; 95% confidence interval, 1.177-2.720; P = .006). Our results suggest that KOR is involved in the carcinogenesis or progression of ESCC and that nuclear KOR may be indicative of prognosis.

Sigma1 receptor (sigma1R), a significant protein, has been found to be frequently upregulated in human tumor cells and tissues. It has been demonstrated that sigma1R is involved in proliferation and adhesion of cancer cells. However, the significance of sigma1R expression in esophageal squamous cell carcinoma (ESCC) remains unclear. In this article, by a series of methods, the authors examined the expression of sigma1R protein in ESCC cell lines and tissues. Flow cytometry indicated intense staining of sigma1R in ESCC cells. Immunocytochemistry staining demonstrated that sigma1R was mainly distributed in cytoplasm and nucleus in ESCC cell lines. Western blotting was performed to characterize the relative expression of sigma1R in different ESCC cell lines. Moreover, different levels of sigma1R were presented from normal epithelium to carcinoma by immunohistochemistry analysis, which demonstrated that sigma1R was highly expressed in tumors. Association analysis showed significant correlations between total sigma1R protein levels and pathologic TNM (pTNM) classification of tumors ( r=0.216, p=0.011). Furthermore, the sigma1R in the nucleus was significantly correlated with pTNM classification and lymph node metastasis ( r=0.263, p=0.002, and r=0.269, p=0.002, respectively). These data indicated that sigma1R may serve as a potential predictive factor for pTNM classification and tumor development in ESCC.

The pyridine extracts of Pingshuo coal were fractionated by silica gel chromatography. Several fractions have been characterized in terms of their average molecular structures using a combination of 1H- and 13C-NMR spectroscopy. The 1H- and 13C-NMR spectra of the fractions revealed distributions of hydrogen as well as carbon in the fractions. From the NMR data, average molecular weight and elemental analysis of the fractions, the average molecular structure parameters were calculated, and their structure models were represented based on these average parameters. Some significant structural differences were found which have provided reliable information in detail for Pingshuo coal bulk structure.

The macromolecular strucutures of the three major macerals, vitrinite, exinite and fusinite, separated from a unique Pingshuo bituminous coal are probed and characterized using advanced analytical techniques including 13C-NMR, XPS, FTIR and XRD. Structural parameters of the individual macerals, for instance, aromaticity, carbon and hydrogen distribution, existing form of heteroatoms and functional groups are measured and investigated. The microcrystallite parameters are also calculated from XRD analysis. The results indicate that the differences in macromolecular structure of the three macerals are obvious and can be distinguished from each other.

The present study was conducted to evaluate the effects of T-2 toxin on semen quality, fertility and serum testosterone concentration in mice. Adult male mice were mated with sexually mature untreated female mice after being exposed to intraperitoneal injection of T-2 toxin at 0, 5, 10 or 15 mg/kg body weight daily for 7 successive days. Semen quality, serum testosterone concentration and fertility of treated mice were assessed. The results showed that the number of abnormal spermatozoa increased significantly and a significant decrease in spermatozoa with integrated acrosome was observed in males treated with T-2 toxin at all doses, As well, the amount of live spermatozoa decreased significantly in mice treated with 10 and 15 mg/kg body weight T-2 toxin. Low pregnancy rate and high fetal resorption rate were observed when females were mated with T-2 toxin—exposed males. Testicular and cauda epididymal sperm counts, efficiency of sperm production and serum testosterone concentration were significantly reduced in mice treated with T-2 toxin at all doses in a dose-dependent manner. In conclusion, these findings indicated that T-2 toxin presented toxic effects on reproductive system of adult male mice.

Seventy-nine rhizobial strains, isolated from root nodules of cowpea ( Vigna unguiculata ) and mung bean (Vigna radiata ) grown in different regions of China, were studied by a fuzzy cluster analysis of 128 phenotypic characteristics. The phenotypic characterization of these strains showed that most of these strains had high stress resistance. For instance, most of them could grow from pH 5.0 to pH 11.0. Over 85% of these strains could grow well on YMA plate at 37 degrees C and several of them even could grow after a 45 minutes hot shock at 60 degrees C. Some strains had a tolerance to high concentration of Bacitracin (400 microg/mL) . The result of the fuzzy cluster analysis showed that all the strains were clustered into 2 groups, slow growers and fast growers, at the similarity level of 63.5% . At the similarity level of 79 %, there were 7 subgroups further separated. Based upon the result of the numerical taxonomy, these strains together with 22 reference stains were analyzed by the 16S rDNA PCR-RFLP. Thirty-four genotype profiles were obtained from the fingerprinting of the 16S rDNA PCR-RFLP. These strains were analyzed by GelCompare II software and clustered into 7 groups at the similarity level of 91% , which were consonant with the 7 subgroups clustered at the similarity level of 79% in numerical taxonomy. The results of numerical taxonomy and 16S rDNA PCR-RFLP analysis showed that all of the seventy-nine rhizobial Bradyrhizobium, strains isolated from root nodules of cowpea and mung bean were clustered into four genera: Agrobacterium, Rhizobium and Sinorhizobium, respectively. An individual clade without any reference stains, which was composed of CCBAU 45071, CCBAU 45111-1 and CCBAU 45248, might be a new species of Rhizobium. Overall, the study results demonstrated a high phenotypic and phylogenetic diversity of rhizobial strains nodulating cowpea and mung bean grown in different geographic regions of China.

Neutrophil gelatinase-associated lipocalin (NGAL) was a novel member of the lipocalin family. The authors previously found that NGAL was overexpressed in the progress of malignant transformation from human immortalized esophageal epithelial cell line SHEE to esophageal carcinoma cell line SHEEC. However, the regulation mechanism of NGAL overexpression was not known. The objective of this study was to clone 5'-untranslated region(5'-UTR) and 3'untranslated region (3'-UTR) of NGAL in SHEEC and to analyze their structural characters.5'-UTR and 3'-UTR of NGAL were cloned from SHEEC using rapid amplification of cDNA ends(RACE). After sequencing the alignment of their nucleotides was analyzed by BLAST database of NCBI and the potential cis-acting elements in the 3'-UTR were identified by computer analysis.The authors cloned and sequenced 69 bp 5'-UTR and 147 bp 3'-UTR of NGAL gene on the basis of the previous works and did not find any base pair mutation.NGAL gene from SHEEC had the entire 5'-UTR and 3'-UTR.