Aberrant splicing in neuroblastoma generates RNA-fusion transcripts and provides vulnerability to spliceosome inhibitors.

The paucity of recurrent mutations has hampered efforts to understand and treat neuroblastoma. Alternative splicing and splicing-dependent RNA-fusions represent mechanisms able to increase the gene product repertoire but their role in neuroblastoma remains largely unexplored. Here we investigate the presence and possible roles of aberrant splicing and splicing-dependent RNA-fusion transcripts in neuroblastoma. In addition, we attend to establish whether the spliceosome can be targeted to treat neuroblastoma. Through analysis of RNA-sequenced neuroblastoma we show that elevated expression of splicing factors is a strong predictor of poor clinical outcome. Furthermore, we identified >900 primarily intrachromosomal fusions containing canonical splicing sites. Fusions included transcripts from well-known oncogenes, were enriched for proximal genes and in chromosomal regions commonly gained or lost in neuroblastoma. As a proof-of-principle that these fusions can generate altered gene products, we characterized a ZNF451-BAG2 fusion, producing a truncated BAG2-protein which inhibited retinoic acid induced differentiation. Spliceosome inhibition impeded neuroblastoma fusion expression, induced apoptosis and inhibited xenograft tumor growth. Our findings elucidate a splicing-dependent mechanism generating altered gene products in neuroblastoma and show that the spliceosome is a potential target for clinical intervention.

SIRT2 Is Critical for Sheep Oocyte Maturation through Regulating Function of Surrounding Granulosa Cells.

Oocyte in vitro maturation is crucial for in vitro embryo production technology, which provides oocytes resources for in vitro fertilization and somatic cell nuclear transfer. Previous studies proved that SIRT2, a member of the sirtuin family, plays a role in oocyte meiosis, but its role in sheep oocyte maturation and its regulating mechanism remains unknown. Firstly, we confirmed the role of Sirt2 in sheep oocytes maturation by supplementation of SIRT2 inhibitor and activator. To further explore the specific mechanism, we performed knockdown of Sirt2 in granulosa cells and then cocultured them with oocytes. Moreover, we determined the effects of Sirt2 on granulosa cell oxidative apoptosis, cell migration, and diffusion, and examined its effects on granulosa cell mitochondrial function, mitophagy, and steroid hormone levels. The results showed that supplementation of SIRT2 inhibitor decreased the oocytes maturation rate (69.28% ± 1.28 vs. 45.74% ± 4.74, p < 0.05), while resveratrol, a SIRT2 activator, increased its maturation rate (67.44% ± 1.68 vs. 78.52 ± 1.28, p < 0.05). Knockdown of Sirt2 in sheep granulosa cells also reduced the oocytes maturation rate (47.98% ± 1.43 vs. 33.60% ± 1.77, p < 0.05), and led to decreased cell migration and expansion ability, oxidative apoptosis, abnormal mitochondrial gene expression, decreased mitochondrial membrane potential and ATP level, and increased mitophagy level. Overexpression of Sirt2 improved mitochondrial membrane potential and ATP level and improved mitochondrial function. Furthermore, we found that Sirt2 knockdown in granulosa cells promotes the secretion of P4 through regulating p-ERK1/2. In conclusion the present study showed that SIRT2 is critical for sheep oocyte maturation through regulating the function of ovarian granulosa cells, especially affecting its mitochondrial function.

Effects of a new fermented soya bean meal on growth performance, serum biochemistry profile, intestinal immune status and digestive enzyme activities in piglets.

BACKGROUND: Fermented soya bean meal (FSBM) is believed to have improved nutritional qualities compared with soya bean meal (SBM) and is also cheaper than soya protein concentration (SPC) and fish meal (FM). Therefore, the present study was conducted to compare the effects of FSBM replacing SBM, SPC and FM in diets on growth performance, serum biochemistry profile, short-chain fatty acid concentrations in digesta, intestinal mucosal enzyme activities, intestinal proinflammatory cytokine concentrations and morphology in weaned piglets. One hundred and twenty 28-day-old piglets (Duroc × Landrace × Yorkshire, body weight: 6.73 ± 1.14 kg) were randomly allocated to four treatment diets (six replicate pens with five piglets per pen) containing SBM, SPC, FM or FSBM as the protein source, respectively. RESULTS: Dietary FSBM supplementation improved average daily gain (p < 0.05), gain to feed ratio (p < 0.05), and digestibility of dry matter, gross energy, crude protein and organic matter (p < 0.05) in pigs compared with those fed SBM during 0-14 days and reduced diarrhoea rate (p < 0.05) compared with those fed SBM and FM during 0-14 days. Moreover, pigs fed FBSM had greater IgA and IgM contents and antioxidase activities than those provided SBM and SPC on day 14. In addition, the butyrate concentration in the cecum of pigs fed FSBM was greater than those fed the other diets (p < 0.05), and the trypsin activity in duodenum and jejunum of pigs provided FSBM was greater than those fed SBM (p < 0.05). Moreover, higher villus height (p < 0.05) and villus height to crypt depth ratio (p < 0.05) and lower crypt depth (p < 0.05) in the duodenum of pigs fed FSBM were observed, and pigs fed FSBM had a lower (p < 0.05) TNF-α concentration in jejunum compared with those fed SBM. CONCLUSIONS: In conclusion, dietary FSBM supplementation to replace SBM, SPC and FM could improve piglets' growth performance, intestinal health and immune function.

The miRNA-mRNA modules enhance juvenile hormone biosynthesis for insect vitellogenesis and egg production.

In addition to preventing precocious larval metamorphosis, juvenile hormone (JH), synthesized in corpora allata (CA), is known to stimulate female reproduction of insects. JH titer is extremely low or absent during metamorphosis, but thereafter rapidly increases in the previtellogenic stage and rises to a peak in the vitellogenic phase. However, the mechanisms underlying the biosynthesis of high levels of JH in adults remain unclear. We found in this study that 12 genes involved in JH synthesis pathway were highly expressed in the CA of adult locusts. By transcriptome analysis and quantitative real-time - polymerase chain reaction validation, a total of 106 evolutionary conserved micro RNAs (miRNAs) and 163 species-specific miRNAs were identified in locust CA. Dual-luciferase assay revealed that 17 miRNAs bound to 10 JH synthesis genes (JHSGs) and downregulated their expression. These miRNAs were expressed in low levels during vitellogenic stage, which was oppositive from that of targeting JHSGs. Six miRNAs including miR-971-3p, miR-31a, miR-9-5p, miR-1-3p, miR-315, and miR-282 were selected for function study. Co-application of agomiRs resulted in significantly decreased levels of targeting JHSGs, accompanied by significantly reduced vitellogenin expression as well as arrested ovarian development. The data suggest that multiple miRNAs expressed synchronously at low levels in the vitellogenic phase, thereby ensuring the high levels of JHSG expression to facilitate JH biosynthesis required for JH-dependent female reproduction. The findings provide important information for deciphering miRNA-messenger RNA modules for JH biosynthesis as well as JH regulation of insect metamorphosis and reproduction.

Advances in the preparation, characterization, and biological functions of chitosan oligosaccharide derivatives: A review.

Chitosan oligosaccharide (COS), which represent the positively charged basic amino oligosaccharide in nature, is the deacetylated and degraded products of chitin. COS has become the focus of intensive scientific investigation, with a growing body of practical and clinical studies highlighting its remarkable health-enhancing benefits. These effects encompass a wide range of properties, including antibacterial, antioxidant, anti-inflammatory, and anti-tumor activities. With the rapid advancements in chemical modification technology for oligosaccharides, many COS derivatives have been synthesized and investigated. These newly developed derivatives possess more stable chemical structures, improved biological activities, and find applications across a broader spectrum of fields. Given the recent interest in the chemical modification of COS, this comprehensive review seeks to consolidate knowledge regarding the preparation methods for COS derivatives, alongside discussions on their structural characterization. Additionally, various biological activities of COS derivatives have been discussed in detail. Lastly, the potential applications of COS derivatives in biomedicine have been reviewed and presented.

High-throughput neural stem cell-based drug screening identifies S6K1 inhibition as a selective vulnerability in sonic hedgehog-medulloblastoma.

BACKGROUND: Medulloblastoma (MB) is one of the most common malignant brain tumors in children. Current treatments have increased overall survival but can lead to devastating side effects and late complications in survivors, emphasizing the need for new, improved targeted therapies that specifically eliminate tumor cells while sparing the normally developing brain. METHODS: Here, we used a sonic hedgehog (SHH)-MB model based on a patient-derived neuroepithelial stem cell system for an unbiased high-throughput screen with a library of 172 compounds with known targets. Compounds were evaluated in both healthy neural stem cells (NSCs) and tumor cells derived from the same patient. Based on the difference of cell viability and drug sensitivity score between normal cells and tumor cells, hit compounds were selected and further validated in vitro and in vivo. RESULTS: We identified PF4708671 (S6K1 inhibitor) as a potential agent that selectively targets SHH-driven MB tumor cells while sparing NSCs and differentiated neurons. Subsequent validation studies confirmed that PF4708671 inhibited the growth of SHH-MB tumor cells both in vitro and in vivo, and that knockdown of S6K1 resulted in reduced tumor formation. CONCLUSIONS: Overall, our results suggest that inhibition of S6K1 specifically affects tumor growth, whereas it has less effect on non-tumor cells. Our data also show that the NES cell platform can be used to identify potentially effective new therapies and targets for SHH-MB.

Assessment of Genetic Diversity in Bupleurum spp. Basing Agronomic Traits, Medicinal Components and ISSR Markers.

Radix bupleuri is one of the bulk medicinal materials in China and it is widely adopted in clinical applications and drug discovery. The investigation of agronomic traits, active component content and genetic diversity in diverse Radix bupleuri germplasms may provide evidence to promote the selection of better strains. In this research, 13 germplasms from various sources were used to investigate the variations between different Radix bupleuri germplasms. Nine biological characteristics were noted in the field, and the levels of the two primary active ingredients were determined using high performance liquid chromatography (HPLC). Moreover, the molecular marker technique of inter-simple sequence repeat (ISSR) and the unweighted pair group method with arithmetic means (UPGMA) were employed to evaluate the molecular genetic diversity. The findings showed that there was a wide range of variation among the many varieties of Radix bupleuri, with coefficients of variation for agronomic traits and active component content ranging from 7.62% to 41.54% and 36.47% to 53.70%, respectively. Moreover, there are different degrees of relationship between the two. Since there was a significant correlation between root weight and saikosaponin content, it was possible to classify a plant based on its weight and anticipate its saikosaponin content. The 13 species were divided into four groups based on their germplasm by genetic markers-based cluster analysis. This indicated the possibility that the component content would not necessarily be related to germplasm and might easily be influenced by environmental factors. The use of ISSR marker technology made it possible to precisely identify the various Radix bupleuri provenances and its counterfeit products. There may be a way to prevent the misunderstandings caused by the appearance and composition of Chinese medicinal substances. In our study, the germplasm of Radix bupleuri that was widely circulated in the market was comprehensively evaluated in terms of agronomic traits, active components and molecular level, and identified by simple means, to provide a theoretical basis for the evaluation and screening of fine germplasms of Radix bupleuri.

Selection and prediction of metro station sites based on spatial data and random forest: a study of Lanzhou, China.

Urban economic development, congestion relief, and traffic efficiency are all greatly impacted by the thoughtful planning of urban metro station layout. with the urban area of Lanzhou as an example, the suitability of the station locations of the built metro stations of the rail transit lines 1 and 2 in the study area have been evaluated using multi-source heterogeneous spatial data through data collection, feature matrix construction, the use of random forest and K-fold cross-validation, among other methods. The average Gini reduction value was used to examine the contribution rate of each feature indicator based on the examination of model truthfulness. According to the study's findings: (1) K-fold cross-validation was applied to test the random forest model that was built using the built metro stations and particular factors. The average accuracy of the tests and out-of-bag data (OOB) of tenfold cross-validation were 89.62% and 91.285%, respectively. Additionally, the AUC area under the ROC curve was 0.9823, indicating that this time, from the perspective of the natural environment, traffic location, and social factors The 19 elements selected from the views of the urban function structure, social economics, and natural environment are closely associated to the locations of the metro station in the research region, and the prediction the findings are more reliable; (2) It becomes apparent that more than half of the built station sites display excellent agreement with the predicted sites in terms of geographical location by superimposing the built metro station sites with the prediction results and tally up their cumulative prediction probability values within the 300 m buffering zone; (3) Based on the contribution rate of each indicator to the model, transport facilities, companies, population density, night lighting, science, education and culture, residential communities, and road network density are identified as the primary influential factors, each accounting for over 6.6%. Subsequently, land use, elevation, and slope are found to have relatively lower contributions. The results of the research provided important information for the local metro's best location selection and planning.

Accumulated Dose Deviation of Rotational and Residual Setup Errors on Nasopharyngeal Carcinoma Using MIM Treated by Helical Tomotherapy.

BACKGROUND: To analyze the relationship between the rotational and residual setup errors and the dose deviation on nasopharyngeal carcinoma (NPC) treated by helical tomotherapy (HT). METHODS: From 25 July 2017 to 20 August 2019, 16 treated NPC patients were enrolled in the study. These patients were scanned with full target range megavoltage computed tomography (MVCT) every other day. Adaptive radiotherapy function application software MIM7.1.3 were used to accumulate the actual dose. The dose deviation with the initial plan dose of the patients' target and organs at risk (OAR) were compared, and the correlation between the dose change and the setup errors (rotational setup errors and neck residual setup error) was analyzed. RESULTS: Translational setup errors increased farther away from the head. Statistically significant difference among 3 groups was achieved in the directions of left-right (P < .001) and anteroposterior (P < .001) by analysis of variance test. Compared with the initial plan dose, the actual accumulated dose of the target area decreased with the actual exposure dose of the OAR increased. However, most of the dosimetric parameters differed by less than 5%. No correlation was found between dose deviation values and the translational setup errors of target. However, sagittal rotational setup errors (pitch) had a positive relationship (P < .05) with the avearge dose of PTVnd (L) (r = 0.885), PTVnd(R) (r = 0.547) PTV1(r = 0.633) and PTV2(r = 0.584). Transverse rotational setup errors (roll) had a positive relationship (P < .05) with the avearge dose of PTVnd(R) (r = 0.593), PTV1(r = 0.505) and PTV2(r = 0.662). CONCLUSIONS: Dose deviation between the actual accumulated and initial plan is not negligible, but most indicators difference is less than 5%, NPC patients treated by HT with MVCT correction setup errors every other day did not need adaptive radiotherapy model unless got rapid tumor shrinkage or weight loss. Moreover, to minimize the dose deviation, more attention should be paid to the reduction of pitch, roll, and residual error of cervical vertebrae during body positioning.

Trophic niche and potential carbon source of three reef-associated fishes of Zhongjieshan Islands.

To clarify the trophic relationship of important rock fishes, we analyzed trophic niche of three typical rockfish species (Oplegnathus fasciatus, Sebastiscus marmoratus and Conger myriaster) in the Zhongjieshan Islands in summer 2020, based on the carbon and nitrogen stable isotope techniques. We calculated the contributions of major carbon sources [macroalgae, phytoplankton, suspended particulate organic matter (POM) and substrate organic matter (SOM)]. The results showed that: 1) the δ13C values of the three species ranged from -21.44‰ to -15.21‰, with an average value of (-16.85±1.12)‰, while the δ15N values ranged from 8.32‰ to 10.96‰, with an average value of (9.69±0.66)‰. There were significant differences in carbon and nitrogen stable isotopes among the three species. 2) There was small niche overlap between O. fasciatus and S. marmoratus, indicating that the interspecific competition was not intense. There was no overlap between C. myriaster and the first two, indicating feeding differentiation. 3) The total ecotone area, corrected core ecotone area, and food source diversity of C. myriaster were the highest, indicating that it had a more generalized diet and richer food sources. 4) With Mytilus coruscus as a baseline organism, the trophic level of C. myriaster was the highest (3.38), followed by S. marmoratus (3.09), and the trophic level of O. fasciatus was the smallest (3.00). 5) Results of the stable isotope mixture model (SIAR) showed that POM was the main carbon source of the three species, contributing 57.4%, 57.9%, and 92.0% of the total, respectively. In addition, the contribution rate of SOM was also high for O. fasciatus and S. marmoratus, which was 21.5% and 33.9%, respectively. This study could provide basic information and reference for understanding trophic structure and marine food web in Zhongjiashan Islands.

An exploratory analysis of MR-guided fractionated stereotactic radiotherapy in patients with brain metastases.

Purpose: To assess the feasibility and potential benefits of online adaptive MR-guided fractionated stereotatic radiotherapy (FSRT) in patients with brain metastases (BMs). Methods and materials: Twenty-eight consecutive patients with BMs were treated with FSRT of 30 Gy in 5 fractions on the 1.5 T MR-Linac. The FSRT fractions employed daily MR scans and the contours were utilized to create each adapted plan. The brain lesions and perilesional edema were delineated on MR images of pre-treatment simulation (Fx0) and all fractions (Fx1, Fx2, Fx3, Fx4 and Fx5) to evaluate the inter-fractional changes. These changes were quantified using absolute/relative volume, Dice similarity coefficient (DSC) and Hausdorff distance (HD) metrics. Planning target volume (PTV) coverage and organ at risk (OAR) constraints were used to compare non-adaptive and adaptive plans. Results: A total of 28 patients with 88 lesions were evaluated, and 23 patients (23/28, 82.1%) had primary lung adenocarcinoma. Significant tumor volume reduction had been found during FSRT compared to Fx0 for all 88 lesions (median -0.75%, -5.33%, -9.32%, -17.96% and -27.73% at Fx1, Fx2, Fx3, Fx4 and Fx5, p < 0.05). There were 47 (47/88, 53.4%) lesions being accompanied by perilesional edema and the inter-fractional changes were significantly different compared to those without perilesional edema (p < 0.001). Patients with multiple lesions (13/28, 46.4%) had more significant inter-fractional tumor changes than those with single lesion (15/28, 53.6%), including tumor volume reduction and anatomical shift (p < 0.001). PTV coverage of non-adaptive plans was below the prescribed coverage in 26/140 fractions (19%), with 12 (9%) failing by more than 10%. All 140 adaptive fractions met prescribed target coverage. The adaptive plans also had lower dose to whole brain than non-adaptive plans (p < 0.001). Conclusions: Significant inter-fractional tumor changes could be found during FSRT in patients with BMs treated on the 1.5 T MR-Linac. Daily MR-guided re-optimization of treatment plans showed dosimetric benefit in patients with perilesional edema or multiple lesions.

Overexpression of miR-651-5p inhibits ultraviolet radiation-induced malignant biological behaviors of sebaceous gland carcinoma cells by targeting ZEB2.

Background: Ultraviolet (UV) exposure is the most essential etiological factor in sebaceous gland carcinoma (SGC). The abnormal expression of microRNAs (miRNAs) is also involved in SGC. However, the function of miRNAs in UV-induced SGC is still unclear. Methods: In this study, the expression levels of miR-651-5p and zinc finger E-box binding homeobox 2 (ZEB2) in SGC tissues and cells were measured by real-time quantitative polymerase chain reaction (RT-qPCR) and western blotting. Then, the effects of miR-651-5p on the apoptosis, migration, invasion, and epithelial-mesenchymal transition (EMT) of UV-induced SGC cells were determined. The interactions between miR-651-5p and ZEB2 were verified by a dual-luciferase reporter assay. An in vivo tumor growth assay was performed to assess tumorigenicity. Results: The results showed that there was abnormal expression of miR-651-5p and ZEB2 in SGC tissues and cells compared with the control tissues and cells. Overexpression of miR-651-5p and knockdown of ZEB2 inhibited the malignant biological behaviors of SGC cells. Moreover, ZEB2 is one of the target genes of miR-651-5p, and the expression of ZEB2 was negatively regulated by miR-651-5p in SGC cells. Further studies showed that overexpression of miR-651-5p promoted cell apoptosis and inhibited the cell invasion and migration ability and EMT of UV-induced SGC cells by downregulating the expression of ZEB2 in vitro and in vivo. Conclusions: This study revealed that overexpression of miR-651-5p inhibited UV-induced SGC growth and metastasis by suppressing ZEB2, which may be a potential target for SGC prevention and therapy.

The complete chloroplast genome sequence of Spiraea×vanhouttei (Briot) Zabel (Rosaceae).

Spiraea×vanhouttei (Rosaceae) is a frequently planted Spiraea species that is distributed in Shandong Province, Jiangsu Province, and Guangdong Province, China. The first complete chloroplast genome of Spiraea×vanhouttei was determined and described in this study. The genome is 155,957 bp in length and contained 129 encoded genes in total, including 84 protein-coding genes, eight ribosomal RNA genes, and 37 transfer RNA genes. The phylogenomic analysis showed that Spiraea×vanhouttei was closely related to Spiraea blumei according to the current sampling extent.

Effects of Selenium Auricularia cornea Culture Supplementation on Growth Performance, Antioxidant Status, Tissue Selenium Concentration and Meat Quality in Growing-Finishing Pigs.

Selenium Auricularia cornea culture (SAC) is a new source of organic selenium. Two experiments were conducted to determine the available energy of SAC fed to pigs and to evaluate the effects of dietary SAC supplementation on growth performance, serum biochemical profiles, fecal short chain fatty acids (SCFA), meat quality, tissue selenium concentration, and oxidative stability of fresh meat in growing-finishing pigs. In Experiment (Exp.) 1, 12 barrows with average body weight (BW) of 42.40 ± 5.30 kg were randomly allotted to two groups and fed the basal diet and SAC-supplemented diet, individually. In Exp. 2, 96 growing-finishing pigs (BW: 91.96 ± 7.55 kg) were grouped into four dietary treatments; each treatment contained six replicates with four pigs per replicate. The four treatments fed a control diet and three experimental diets supplemented with 0.6%, 1.2%, and 2.4% SAC, respectively. The trial lasted for 45 days. The results revealed that digestible energy (DE) of SAC was 11.21 MJ/kg. The average daily gain (ADG) was improved in pigs fed 1.2% and 2.4% SAC during day 24 to 45 and the overall period. Dietary 1.2% and 2.4% SAC supplementation had a lower F/G (p < 0.05) than the control diet during different stages. Dietary SAC supplementation increased fecal butyrate contents (p < 0.05), and pigs fed 1.2% and 2.4% SAC diets had a higher MCT1 mRNA expression (p = 0.04) in the colon. Pigs fed 2.4% SAC had higher GSH-Px contents (p < 0.05) in serum, liver, and longissimus dorsi muscle (LDM) than those in the control group. The 2.4% SAC-supplemented group revealed a higher Se content (p < 0.05) in LDM and a lower MDA concentration (p < 0.05) in fresh meat during the simulated retail display on day six. In conclusion, this study suggested that SAC was more effective in improving growth, enhancing the antioxidant status, depositing Se in muscle, and increasing meat oxidative stability of pigs.

Transcriptome profiling of Ewing sarcomas - treatment resistance pathways and IGF-dependency.

Ewing sarcomas (ESs) are aggressive sarcomas driven by EWS fusion genes. We sought to investigate whether whole-transcriptome sequencing (RNA-seq) could be used to detect patterns associated with chemotherapy response or tumor progression after first-line treatment. Transcriptome sequencing (RNA-seq) of 13 ES cases was performed. Among the differentially expressed pathways, we identified IGF2 expression as a potential driver of chemotherapy response and progression. We investigated the effect of IGF2 on proliferation, radioresistance, apoptosis, and the transcriptome pattern in four ES cell lines and the effect of IGF2 expression in a validation series of 14 patients. Transcriptome analysis identified differentially expressed genes (adj. P < 0.005) and pathways associated with chemotherapy response (285 genes), short overall survival (662 genes), and progression after treatment (447 genes). Imprinting independent promoter P3-mediated IGF2 expression was identified in a subset of cases with aggressive clinical course. In ES cell lines, IGF2 induced proliferation, but promoted radioresistance only in CADO cells. High IGF2 expression was also significantly associated with shorter overall survival in patients with ES. Transcriptome analysis of the clinical samples and the cell lines revealed an IGF-dependent signature, potentially related to a stem cell-like phenotype. Transcriptome analysis is a potentially powerful complementary tool to predict the clinical behavior of ES and may be utilized for clinical trial stratification strategies and personalized oncology. Certain gene signatures, for example, IGF-related pathways, are coupled to biological functions that could be of clinical importance. Finally, our results indicate that IGF inhibition may be successful as a first-line therapy in conjunction with conventional radiochemotherapy for a subset of patients.

Nuclear IGF1R interact with PCNA to preserve DNA replication after DNA-damage in a variety of human cancers.

Nuclear IGF1R has been linked to poor outcome in cancer. We recently showed that nuclear IGF1R phosphorylates PCNA and increases DNA damage tolerance. In this paper we aimed to describe this mechanism in cancer tissue as well as in cancer cell lines. In situ proximity ligation assay identified frequent IGF1R and PCNA colocalization in many cancer types. IGF1R/PCNA colocalization was more frequently increased in tumor cells than in adjacent normal, and more prominent in areas with dysplasia and invasion. However, the interaction was often lost in tumors with poor response to neoadjuvant treatment and most metastatic lesions. In two independent cohorts of serous ovarian carcinomas and oropharyngeal squamous cell carcinomas, stronger IGF1R/PCNA colocalization was significantly associated with a higher overall survival. Ex vivo irradiation of ovarian cancer tissue acutely induced IGF1R/PCNA colocalization together with γH2AX-foci formations. In vitro, RAD18 mediated mono-ubiquitination of PCNA during replication stress was dependent on IGF1R kinase activity. DNA fiber analysis revealed that IGF1R activation could rescue stalled DNA replication forks, but only in cancer cells with baseline IGF1R/PCNA interaction. We believe that the IGF1R/PCNA interaction is a basic cellular mechanism to increase DNA stress tolerance during proliferation, but that this mechanism is lost with tumor progression in conjunction with accumulated DNA damage and aberrant strategies to tolerate genomic instability. To exploit this mechanism in IGF1R targeted therapy, IGF1R inhibitors should be explored in the context of concomitant induction of DNA replication stress as well as in earlier clinical stages than previously tried.

Balancing carbon/nitrogen ratio to improve nutrients removal and algal biomass production in piggery and brewery wastewaters.

To improve nutrients removal from wastewaters and enhance algal biomass production, piggery wastewater was mixed with brewery wastewaters. The results showed that it was a promising way to cultivate microalga in piggery and brewery wastewaters by balancing the carbon/nitrogen ratio. The optimal treatment condition for the mixed piggery-brewery wastewater using microalga was piggery wastewater mixed with brewery packaging wastewater by 1:5 at pH 7.0, resulting in carbon/nitrogen ratio of 7.9, with the biomass concentration of 2.85 g L-1, and the removal of 100% ammonia, 96% of total nitrogen, 90% of total phosphorus, and 93% of chemical oxygen demand. The application of the established strategies can enhance nutrient removal efficiency of the wastewaters while reducing microalgal biomass production costs.

Characterization of additional zinc ions on the growth, biochemical composition and photosynthetic performance from Spirulina platensis.

In this study, the effect of various initial Zn2+ concentration additionally on microalgae growth and biochemical composition were investigated. The Spirulina platensis biomass of each concentration reached the maximum at the end of the cultivation. However, high levels could severely inhibit the growth of microalgae. Fluorescence activity occurred changes in response to heavy metal stress. Moreover, biochemical composition in Spirulina platensis altered under zinc stress, and the highest contents of phycocyanin (PC), Chlorophyll-a (Chl-a), Carotenoid and zinc accumulation were obtained. The proportion of saturated and polyunsaturated fatty acids increased constantly in response to Zn2+ exposure. Overall, this study indicated that the use of Spirulina platensis is a viable method for treating zinc containing wastewater and harvested microalgae can be processed into high-zinc products.

CuInS2/ZnS QD exposure induces developmental toxicity, oxidative stress and DNA damage in rare minnow (Gobiocypris rarus) embryos and larvae.

Chinese rare minnow (Gobiocypris rarus) embryos were used as an experimental model to investigate the effects of CuInS2/ZnS quantum dots (QDs) on the early life stages of G. rarus. Normal developmental parameters (survival rate, body length and average heart rate), biomarker genes [stress response (Hsp70), detoxification (Cyp1a), organizer function and axis formation (Wnt8α), and muscle (Mstn)], enzymatic activity and DNA damage were recorded as endpoints in the developing embryos/larvae after exposure until 96h post-fertilization (hpf). Reduced survival rate, decreased heart rate, altered body length, increased malformation rate, decreased hatching rate, advanced hatching time in response to low concentrations (50 and 100nmol/L) and delayed hatching time in response to high concentrations were observed after exposure, as were many other toxic effects, including pericardial edema and bent tails. The 72 hpf LC50 (median lethal concentration) was determined to be 624.364nmol/L. Treatment with certain concentrations of CuInS2/ZnS QDs significantly increased the superoxide dismutase (SOD) activity and malondialdehyde (MDA) levels and significantly induced DNA damage. After treatment with CuInS2/ZnS QDs, the embryos showed highly up-regulated expression of Hsp70, Cyp1a and Wnt8a and significantly up-regulated expression of Mstn at 12 hpf. Overall, this study indicates that CuInS2/ZnS QDs are potentially toxic to G. rarus embryos. The information presented in this study will be helpful for fully understanding the toxicity induced by CuInS2/ZnS QDs in fish embryos.

Developmental Toxicity of Carbon Quantum Dots to the Embryos/Larvae of Rare Minnow (Gobiocypris rarus).

The toxic effects of CDs on rare minnow (Gobiocypris rarus) embryos at different developmental stages were investigated. The results showed that rare minnow embryos had decreased spontaneous movements, body length, increased heart rate, pericardial edema, yolk sac edema, tail/spinal curvature, various morphological malformations, and decreased hatching rate. Biochemical analysis showed the CDs exposure significantly inhibited the activity of Na+/K+-ATPase and Ca2+-ATPase and increased the MDA contents and the activity of SOD, CAT, and GPX. Further examination suggested that the CDs exposure induced serious embryonic cellular DNA damage. Moreover, the CDs exposure induced upregulation of development related genes (Wnt8a and Mstn) along with the downregulation of Vezf1. Overall, the present study revealed that the CDs exposure has significant development toxicity on rare minnow embryos/larvae. Mechanistically, this toxicity might result from the pressure of induced oxidative stress coordinate with the dysregulated development related gene expression mediated by the CDs exposure.