Clinical characteristics and prognosis of different primary tumor location in colorectal cancer: a population-based cohort study.

PURPOSE: Emerging data have shown that patients with left-sided cancers have better survival than patients with right-sided cancers in terms of metastatic colorectal cancer. However, the available information and findings remain limited and contradictory in localized colorectal cancer. This study aimed to evaluate the clinical characteristics and prognosis of primary tumor location (PTL) in colorectal cancer. METHODS: Patients' diagnoses were identified using the Surveillance, Epidemiology, and End Result database between 2006 and 2015. The analyses were further stipulated to patients with primary cancer site, histology, and stage information. The correlations between PTL and overall survival (OS) were assessed. RESULTS: Compared with left-sided tumors, right-sided tumors were more likely to develop into T3 cancers (50.0% vs. 44.8%), T4 cancers (15.8% vs. 12.3%), mucinous or mucin-producing adenocarcinoma (10.8% vs. 5.0%), and signet ring cell carcinoma (1.4% vs. 0.7%), P < 0.01, respectively. Patients with right-sided tumors showed inferior OS (56.1% vs. 60.2%), and the hazard ratio was 1.224 (95% CI, 1.208-1.241, P < 0.001) in all stages. Stage-specific Cox regression analysis revealed that patients with right-sided tumors also showed inferior OS in every stage (respectively, P < 0.05) than left-sided tumors. CONCLUSIONS: This study demonstrated that the prognoses of patients with left-sided cancers were better than those of patients with right-sided cancers regardless of stage. PTL can be a prognosis factor in colorectal cancer. We encourage developing clinical and translational studies to elucidate the causative relationship between PTL and prognosis.

Cloning and expression of class I chitinases in Hami melon after Penicillium infection.

Chitinases are important disease-related proteins that play critical roles in plant defense against disease. To investigate the function of chitinases in the resistance of Hami melon to Penicillium infection, the gene encoding chitinases, HmCHT-2, was cloned and RT-PCR was used to measure expression levels of HmCHT-2. When the Hami melon was infected by Penicillium sp after 0, 12, 36, 48, 60, and 72 h. The results showed that comparing to the control group, the time of expression levels reaching to the peak delayed and the expression levels maintained at a significantly high level for a longer time. These results suggest that HmCHT-2 may contribute to the defense of Hami melon against fungal infection.

Molecular cloning and expression analysis of Fem1b from oriental river prawn Macrobrachium nipponense.

Feminization-1 homolog b (Fem1b) is one of the genes essential for male development and play central roles in sex determination of Caenorhabditis elegans. In this study, we cloned and characterized the full-length Fem1b cDNA from the freshwater prawn Macrobrachium nipponense (MnFem1b) in different tissues and at different developmental stages. Real-time quantitative reverse polymerase chain reaction (RT-qPCR) showed that the MnFem1b gene was expressed in all investigated tissues, with the highest expression level found in the testes. The results revealed that the MnFem1b gene might play roles in aspects of development of the male prawn phenotype. The RT-qPCR also revealed that MnFem1b mRNA expression was significantly increased at 10 days after metamorphosis. The expression levels in all investigated tissues showed a certain degree of sexually dimorphism, the expression levels in males were significantly higher than those in females (P < 0.05). Notably, the highest expression of MnFem1b was found in the testes. The expression of MnFem1b in different tissues indicates that it plays multiple biological functions in M. nipponense.

Prognostic value of ZFP36 and SOCS3 expressions in human prostate cancer.

PURPOSE: ZFP36 ring finger protein (ZFP36) and the suppressor of cytokine signaling 3 (SOCS3) have been reported to, respectively, regulate NF-κB and STAT3 signaling pathways. To better understand the correlation of NF-κB and STAT3 negative regulates pathway, we have investigated the involvement of ZFP36 and SOCS3 expressions in human prostate cancer (PCa). METHODS: In the present study, paired patient tissue microarrays were analyzed by immunohistochemistry, and the ZFP36 protein expression was quantitated as immunoreactive scores in patients with PCa. Associations between ZFP36/SOCS3 expression and various clinicopathological features and prognosis of PCa patients were statistically analyzed based on the Taylor database. Then, the functions of ZFP36 and SOCS3 in cancerous inflammation were determined using qPCR and immunohistochemistry in vitro and in vivo. RESULTS: ZFP36 protein expression in PCa tissues was significantly lower than those in non-cancerous prostate tissues (P < 0.05). In mRNA level, ZFP36 and SOCS3 had a close correlation with each other (P < 0.01, Pearson r = 0.848), and its upregulation was both significantly associated with low Gleason score (P < 0.001 and P < 0.001, respectively), negative metastasis (P < 0.001 and P < 0.001, respectively), favorable overall survival (P < 0.001 and P < 0.05, respectively), and negative biochemical recurrence (P < 0.001 and P < 0.001, respectively). Functionally, LPS treatment could lead to the overexpression of ZFP36 and SOCS3 in vitro and vivo. CONCLUSIONS: Our data offer the convincing evidence for the first time that the aberrant expressions of ZFP36 and SOCS3 may be involved into the progression and patients' prognosis of PCa, implying their potentials as candidate markers of this cancer.

Genetic diversity of cultivated and wild tomatoes revealed by morphological traits and SSR markers.

In the current study, morphological traits and molecular markers were used to assess the genetic diversity of 29 cultivated tomatoes, 14 wild tomatoes and seven introgression lines. The three components of the principal component analysis (PCA) explained 78.54% of the total morphological variation in the 50 tomato genotypes assessed. Based on these morphological traits, a three-dimensional PCA plot separated the 50 genotypes into distinct groups, and a dendrogram divided them into six clusters. Fifteen polymorphic genomic simple- sequence repeat (genomic-SSR) and 13 polymorphic expressed sequence tag-derived SSR (EST-SSR) markers amplified 1115 and 780 clear fragments, respectively. Genomic-SSRs detected a total of 64 alleles, with a mean of 4 alleles per primer, while EST-SSRs detected 52 alleles, with a mean of 4 alleles per primer. The polymorphism information content was slightly higher in genomic-SSRs (0.49) than in EST-SSRs (0.45). The mean similarity coefficient among the wild tomatoes was lower than the mean similarity coefficient among the cultivated tomatoes. The dendrogram based on genetic distance divided the 50 tomato genotypes into eight clusters. The Mantel test between genomic-SSR and EST-SSR matrices revealed a good correlation, whereas the morphological matrices and the molecular matrices were weakly correlated. We confirm the applicability of EST-SSRs in analyzing genetic diversity among cultivated and wild tomatoes. High variability of the 50 tomato genotypes was observed at the morphological and molecular level, indicating valuable tomato germplasm, especially in the wild tomatoes, which could be used for further genetic studies.

Effects and mechanism of lipoic acid on beta-amyloid-intoxicated C6 glioma cells.

ß-amyloid peptides (Aßs) can exert neurotoxic effects through induction of oxidative damage, whereas lipoic acid (LA), a powerful antioxidant, can alleviate oxidative damage. In this study, we explored the effect and mechanism of action of LA on beta-amyloid-intoxicated C6 glioma cells. Cells were randomly divided into three groups: control (vehicle), Aß, and LA + Aß. The LA + Aß group was treated with LA for 2 h, then both the Aß-only and the LA + Aß groups were incubated with 25 µM Aß for 24 h. Cell viability was measured by the MTT method. Mitochondrial reduced glutathione (GSH) and oxidized glutathione (GSSG) levels were detected by enzyme-linked immunosorbent assay (ELISA), and the GSH to GSSG ratio calculated. Real-time polymerase chain reaction and western blot analyses were used to detect MnSOD mRNA and protein, respectively. Aß significantly inhibited C6 cell proliferation compared with the control group (P < 0.05). LA markedly increased cell viability compared with the Aß group (P < 0.05). The increased GSSH and decreased GSH mitochondrial accumulation induced by Aß was profoundly reversed by treatment with LA (P < 0.05). Aß significantly reduced MnSOD expression compared to controls (P < 0.05), whereas LA pretreatment increased MnSOD expression compared with the Aß-only group (P < 0.05); MnSOD protein levels showed similar patterns. These results suggest that LA might protect Aß-intoxicated C6 glioma cells by alleviating oxidative damage, providing a new treatment strategy for neurodegenerative diseases.

Exceptional material requirement for reproduction in mouse oocytes.

Limited information on oocytes and fertilization prevents the efficient therapy of patients with infertility. The most important reason for this lack of understanding is a deficiency in research dedicated to oocytes and fertilization. Currently, we are concerned with the role of nutrition in the process of oocyte development to better understand the relationship between nutrition and infertility. The aim of this study was to explore the relationship between some exceptional materials and infertility to elucidate the role of these materials in oocyte development. We used proteomic analysis to identify numerous nutrition-related proteins in three developmental stages: the germinal vesicle stage, the metaphase II-arrested stage, and the fertilized oocyte-zygote stage. Specific proteins were abundantly expressed during the three stages. These proteins included astacin-like metalloendopeptidase, selenium-binding proteins, and other proteins involved in metabolic and signaling pathways. Other proteins were involved in the citrate cycle, the electron transport chain, the urea cycle, fatty acid metabolism, and the insulin signaling pathway. Almost all these proteins exhibited different expression levels in the three stages. The results of the present study provide a better understanding of the molecular mechanisms of early embryonic development and suggest new treatment methods for infertility.

Comparison of gSSR and EST-SSR markers for analyzing genetic variability among tomato cultivars (Solanum lycopersicum L.).

In order to study genetic variability and develop better strategies for the utilization of 48 tomato cultivars from America, China, the Netherlands, and Portugal, genomic simple sequence repeat (gSSR) and EST-derived SSR (EST-SSR) markers were applied. In all, 15 of 82 gSSR and 18 of 115 EST-SSR markers showed polymorphic loci. There were 995 and 2072 clear fragments amplified by polymorphic gSSR and EST-SSR markers, respectively. The total and average number of alleles detected by EST-SSRs (75, 4.2) was more than gSSRs (54, 3.6) as a result of some multi-locus EST-SSRs. A lower polymorphism information content value was found in gSSRs (0.529) compared to EST-SSRs (0.620). Similarity coefficient matrixes of the 48 tomato cultivars were established based on the gSSRs and EST-SSRs, and UPGMA dendrograms were constructed from the gSSRs and EST-SSRs similarity coefficient matrixes. A high similarity was observed between the gSSRs and EST-SSRs dendrograms. Genetic variability of four tomato populations from different countries showed that the observed number of alleles and Nei's genetic diversity were highest in the American population, and the effective number of alleles was highest in the Dutch population. The estimated genetic structure showed some tomato cultivars from different countries shared a common genetic background, which might be related to gene flow. It was inferred that both gSSR and EST-SSR markers were effective to assess genetic variability of tomato cultivars, and the combination of both markers could be more effective for genetic diversity analysis in tomato.

Distribution and localization of abnormally expressed OPTN proteins in RGC5 retinal ganglion cells and their effects on subcellular morphology.

The objectives of this study were to investigate the distributions of abnormally expressed optineurin (OPTN) proteins in retinal ganglion cells (RGC5s) of transgenic rats and their effects on subcellular morphological structures. Green fluorescent protein labeled EGFP wild-type (OPTN(WT)), E50K mutant type (OPTN(E50K)), and OPTN siRNA (si-OPTN) eukaryotic expression plasmids were constructed and transfected into RGC5s. Intracellular structures were labeled with organelle specific fluorescent dyes. Construct localization and cell morphologies were visualized by confocal fluorescence microscopy. OPTN(WT) was observed to be distributed as fine punctate fluorescent particles in the cytoplasm around the nucleus, along with exhibiting nuclear expression. OPTNE50K exhibited similar distribution but with non-uniform fluorescence particle size. si-OPTN distribution was similar to that of EGFP: uniform across the cytoplasm and nucleus. Compared with the negative control group, OPTN(WT), and OPTN(E50K) and to a lesser degree pEGFP-transfected cells exhibited fracture and loss of myofilament proteins and mitochondrial swelling and cytoplasmic accumulation, along with abnormal lysosomal distribution and increased volume, and Golgi fragmentation. However, si- OPTN transfected cells exhibited no significant damage. Therefore, we demonstrated that the E50K mutation disrupts the uniformity of OPTN protein distribution upon exogenous overexpression. Furthermore, these results suggested that si-OPTN transfection, and thus potentially OPTN knockdown, did not impact subcellular morphology of RGC5 cells, whereas transfection, especially when combined with wild-type or mutant OPTN expression, led to substantial abnormalities in subcellular morphological structures. These findings lay a foundation for further research into the function of the OPTN protein.

-173G/C polymorphism in the promoter of MIF is associated with hepatitis B virus infection in a Chinese Han population.

In addition to the host immune response, genetic and environmental factors play crucial roles in the manifestation of hepatitis B virus (HBV) infection. The macrophage migration inhibitory factor (MIF) -173G/C polymorphism (rs755622), located in the promoter region of MIF, may play integral roles in diverse processes, including the immune response. Thus, the MIF -173G/C polymorphism may influence the immune response to HBV during natural infection. We investigated whether the MIF -173G/C polymorphism was associated with susceptibility to HBV infection in a Chinese Han population. A total of 596 HBV infection cases and 612 age-matched controls were recruited for the study. Genotyping of the MIF -173G/C polymorphism was performed using the allele-specific polymerase chain reaction method. The frequencies of the alleles and genotypes in patients and controls were compared using the χ(2) test. Carriers of the variant C allele in MIF -173 G/C were at significantly higher risk of HBV infection than carriers of the wild-type allele (P = 0.032, odds ratio = 0.799, 95% confidence interval = 0.651-0.981). However, there was no significant difference in the distribution of MIF -173G/C genotypes between case and control groups in either population (P = 0.096, degrees of freedom = 2). Our findings indicate that the G to C base change in MIF -173 G/C confers an increased risk of development of HBV infection by altering the expression of MIF in our Chinese Han population.

Effects of abnormal optineurin expression on the survival of the rat retinal ganglion cell line RGC-5.

The OPTN gene is thought to be associated with certain types of glaucoma and the function of the protein for which it encodes, optineurin, has been extensively researched, but with contradictory results. We explored the effects of abnormal optineurin expression on the survival of the rat retinal ganglion cell line RGC-5. Plasmids expressing wild-type (WT) or E50K mutant optineurin, or OPTN-specific double-hairpin small interfering RNA (si-RNA), were transfected into RGC-5 cells. The effects on cell survival were monitored by observation of cell morphology and propidium iodide and Hoechst 33342 fluorescent staining, while expression of optineurin was visualized by fluorescence microscopy. Abnormal optineurin expression influenced the survival of RGCs in vitro, as apoptosis was induced by increased WT and E50K mutant optineurin, while a reduction in apoptosis was observed in cells transfected with OPTN-siRNA. Similar results were also observed in transfected cells treated with apoptotic stimuli. Overexpression of WT and mutant E50K protein resulted in greater cell death, while downregulation decreased RGC-5 apoptosis.

Genomic cloning, expression, and single nucleotide polymorphism association analysis of the insulin-like androgenic gland hormone gene in the oriental river prawn (Macrobrachium nipponense).

Increasing evidence suggests that the insulin-like androgenic gland hormone (IAG) gene plays an important role in male sexual differentiation, metabolism, and growth in crustaceans. In the present study, we isolated the full-length genome sequence of IAG by genome walking based on the cDNA sequence in Macrobrachium nipponense. Four novel single nucleotide polymorphisms (SNPs) were studied, including 509G>T, 529G>T, 590A>T in intron 1, and 2226A>G in intron 2. The association of genetic variation with growth traits [body length (BL) and body weight (BW)] was analyzed. Individuals with GG geno- type at locus 2226A>G maintained higher mean BL (P < 0.01) and BW (P < 0.05) than AA and GA individuals. These results suggest that IAG SNPs may be useful molecular markers for selecting growth traits in M. nipponense.

Partial least squares-based gene expression analysis in preeclampsia.

Preeclampsia is major cause of maternal and fetal morbidity and mortality. Currently, the etiology of preeclampsia is unclear. In this study, we investigated differences in gene expression between preeclampsia patients and controls using partial least squares-based analysis, which is more suitable than routine analysis. Expression profile data were downloaded from the Gene Expression Omnibus database. A total of 503 genes were found to be differentially expressed, including 248 downregulated genes and 255 overexpressed genes. Network analysis identified 5 hub genes, including UBB, PIK3R1, MAPRE1, VEGFA, and ITGB1. Three of these, PIK3R1, VEGFA, and ITGB1, are known to be associated with preeclampsia or preeclampsia-related biological processes. Our findings shed light on expression signatures of preeclampsia patients that can be used as theoretical support in future therapeutic studies.

Caffeic acid improves cell viability and protects against DNA damage: involvement of reactive oxygen species and extracellular signal-regulated kinase

Hormesis is an adaptive response to a variety of oxidative stresses that renders cells resistant to harmful doses of stressing agents. Caffeic acid (CaA) is an important antioxidant that has protective effects against DNA damage caused by reactive oxygen species (ROS). However, whether CaA-induced protection is a hormetic effect remains unknown, as is the molecular mechanism that is involved. We found that a low concentration (10 μM) of CaA increased human liver L-02 cell viability, attenuated hydrogen peroxide (H2O2)-mediated decreases in cell viability, and decreased the extent of H2O2-induced DNA double-strand breaks (DSBs). In L-02 cells exposed to H2O2, CaA treatment reduced ROS levels, which might have played a protective role. CaA also activated the extracellular signal-regulated kinase (ERK) signal pathway in a time-dependent manner. Inhibition of ERK by its inhibitor U0126 or by its specific small interfering RNA (siRNA) blocked the CaA-induced improvement in cell viability and the protective effects against H2O2-mediated DNA damage. This study adds to the understanding of the antioxidant effects of CaA by identifying a novel molecular mechanism of enhanced cell viability and protection against DNA damage.

A novel random amplified polymorphic DNA-based strategy for genetic diversity analysis and identification of tomatoes.

Cultivar identification diagrams (CIDs) provide a rapid and efficient approach for identifying cultivars based on random amplified polymorphic DNA (RAPD) markers. In this paper, 64 tomato cultivars were identified using a CID. Using RAPD profiles, clustering analysis was performed to analyze genetic diversity. The results showed that 8 RAPD primers could completely separate the 64 cultivars according to the obtained polymorphic bands; a CID of the 64 tomato cultivars was then constructed. As verification of the CID validity, 8 randomly selected cultivars were investigated and proven to be well distinguished. In addition, 33 DNA bands were obtained, 20 (60.6%) of which were polymorphic. Genetic distances were calculated with a range of 0.032 to 1.402. Clustering analysis showed that the 64 tomato cultivars were divided into 4 groups with a similarity coefficient of 0.40. Using this novel strategy, with the same RAPD data, both CID and clustering analysis can simultaneously determine tomato cultivars and their genetic diversity.

Caffeic acid improves cell viability and protects against DNA damage: involvement of reactive oxygen species and extracellular signal-regulated kinase.

Hormesis is an adaptive response to a variety of oxidative stresses that renders cells resistant to harmful doses of stressing agents. Caffeic acid (CaA) is an important antioxidant that has protective effects against DNA damage caused by reactive oxygen species (ROS). However, whether CaA-induced protection is a hormetic effect remains unknown, as is the molecular mechanism that is involved. We found that a low concentration (10 µM) of CaA increased human liver L-02 cell viability, attenuated hydrogen peroxide (H2O2)-mediated decreases in cell viability, and decreased the extent of H2O2-induced DNA double-strand breaks (DSBs). In L-02 cells exposed to H2O2, CaA treatment reduced ROS levels, which might have played a protective role. CaA also activated the extracellular signal-regulated kinase (ERK) signal pathway in a time-dependent manner. Inhibition of ERK by its inhibitor U0126 or by its specific small interfering RNA (siRNA) blocked the CaA-induced improvement in cell viability and the protective effects against H2O2-mediated DNA damage. This study adds to the understanding of the antioxidant effects of CaA by identifying a novel molecular mechanism of enhanced cell viability and protection against DNA damage.

Correlation and interventional embolization therapy of posterior intercostal arteries-induced hemoptysis.

The incidence of posterior intercostal arteries-induced hemoptysis, its correlation with primary diseases, and the value of interventional embolization therapy were investigated. Clinical data, multislice spiral computed tomography (MSCT), digital subtraction angiography (DSA), and other imaging data of 143 cases of hemoptysis were retrospectively analyzed. After the offending vessels were subjected to interventional embolization therapy, patients were followed-up for observations of clinical efficacies and complications. Thirty-one patients (21.7%) showed 65 branches of posterior intercostal arteries as the non-bronchial systemic arteries involved in hemoptysis; pleural thickening was evident in 25 (80.6%) cases. Posterior intercostal arteries-induced hemoptysis was observed in 16 of the 27 (59.3%) patients with pulmonary tuberculosis, and in 9 of the 10 (90.0%) patients with pulmonary tuberculosis and pulmonary damage. Posterior intercostal arteries-induced hemoptysis was correlated to pleural thickening (P<0.05), which differed significantly among different underlying diseases (P<0.05). Twenty-eight cases of 58 branches of posterior intercostal arteries were found to be involved in hemoptysis by preoperative chest CT angiogram (CTA); the intraoperative matching rates were 90.3% (28/31) and 89.2% (58/65), respectively. Thirty-one patients received transcatheter arterial embolization (TAE), of which 29 (93.5%) showed immediate hemostasis; 1 case had surgical treatment for ineffectuality, and 2 cases showed recurrence without serious complications. The posterior intercostal arteries were commonly involved in hemoptysis, and were closely associated with pleural thickening and pulmonary tuberculosis, especially when accompanied by pulmonary damage. Complete TAE could improve the treatment effect of hemoptysis and preoperative chest CTA was helpful for interventional embolization therapy.

Enhanced expression of IMPDH2 promotes metastasis and advanced tumor progression in patients with prostate cancer.

PURPOSE: Our previous study showed the upregulation of inosine 5'-monophosphate dehydrogenase type II (IMPDH2) protein in human prostate cancer (PCa) tissues and sera compared to non-cancerous controls by proteomics and ELISA analyses. However, the clinical significance of IMPDH2 in PCa has not been fully elucidated. Thus, the aim of the current study was to investigate the associations of IMPDH2 upregulation with tumor progression in patients with PCa. METHODS: IMPDH2 expression at mRNA and protein levels in human PCa and non-cancerous prostate tissues was respectively detected by qRT-PCR, Western blot and immunohistochemistry analyses, which was validated by microarray-based Taylor Data. Then, the association of IMPDH2 expression with clinicopathological features of PCa patients was statistically analyzed. RESULTS: Compared with non-cancerous prostate tissues, IMPDH2 mRNA and protein expression levels were both significantly upregulated (at mRNA level: 9.22 ± 2.49 vs 5.06 ± 1.45, P < 0.01; at protein level by Western blot: 0.674 ± 0.029 vs 0.418 ± 0.140, P < 0.001; at protein level by immunohistochemistry: 4.97 ± 0.760 vs 3.32 ± 1.66, P < 0.001) in PCa tissues, which were consistent with our previous data. In addition, the enhanced expression of IMPDH2 in PCa tissues was significantly correlated with the advanced clinical stage (for our cohort: P < 0.001; for Taylor data: P = 0.002), the presence of metastasis (for our cohort: P < 0.001; for Taylor data: P = 0.012) and the higher Gleason score (for our cohort: P = 0.002; for Taylor data: P = 0.028). CONCLUSIONS: These findings suggest for the first time that the enhanced expression of IMPDH2 may promote the tumor metastasis and the advanced tumor progression in patients with PCa.