Role of Halloween genes in ecdysteroids biosynthesis of the swimming crab (Portunus trituberculatus): Implications from RNA interference and eyestalk ablation.

Molting, including metamorphosis molting in arthropods are controlled by the ecdysteroids that are synthesized and secreted by the crustacean Y-organ (YO) or the insect prothoracic gland (PG). The Halloween genes encoding the enzymes mainly involved in the biosynthesis of ecdysteroids are well studied in insects but not in crustaceans. Given the importance of Halloween genes in ecdysteroids biosynthesis, we have previously reported the cDNA cloning of disembodied (Dib) in P. trituberculatus. Here, cDNA sequences of another two Halloween genes, Spook (Spo) and Shadow (Sad), were further identified and characterized. The predicted amino acid sequences for these two Halloween genes of Portunus trituberculatus were compared to those of several other arthropods, and several typical domains of the cytochrome P450 mono-oxygenase (CYP) were identified. Similar to the tissue distribution of Dib, the Spo and Sad also showed high specificity to the YO. RNA interference (RNAi) of these 3 genes indicated they all play essential role in ecdysteroids biosynthesis. To investigate the relationships of the Halloween genes to the eyestalk neuropeptides such as molt-inhibiting hormone (MIH), effects of eyestalk ablation (ESA) on the expression of Dib, Spo and Sad were detected. Expression of Dib and Sad, but not Spo, was significantly induced by ESA. The result indicated that the inhibition of MIH in ecdysteroids biosynthesis may be partly through the transcriptional regulation of certain Halloween genes, such as Dib and Sad, while the Spo might not be the target for MIH signal.

Genomic analysis of immunogenic cell death-related subtypes for predicting prognosis and immunotherapy outcomes in glioblastoma multiforme.

Immunogenic cell death (ICD), a unique form of cancer cell death, has therapeutic potential in anti-tumour immunotherapy. The aim of this study is to explore the predictive potential of ICD in the prognosis and immunotherapy outcomes of glioblastoma multiforme (GBM). RNA sequencing data and clinical information were downloaded from three databases. Unsupervised consistency clustering analysis was used to identify ICD-related clusters and gene clusters. Additionally, the ICD scores were determined using principal component analysis and the Boruta algorithm via dimensionality reduction techniques. Subsequently, three ICD-related clusters and three gene clusters with different prognoses were identified, with differences in specific tumour immune infiltration-related lymphocytes in these clusters. Moreover, the ICD score was well differentiated among patients with GBM, and the ICD score was considered an independent prognostic factor for patients with GBM. Furthermore, two datasets were used for the external validation of ICD scores as predictors of prognosis and immunotherapy outcomes. The validation analysis suggested that patients with high ICD scores had a worse prognosis. Additionally, a higher proportion of patients with high ICD scores were non-responsive to immunotherapy. Thus, the ICD score has the potential as a biomarker to predict the prognosis and immunotherapy outcomes of patients with GBM.

The protective role of circ_0016760 downregulation against sevoflurane­induced neurological impairment via modulating miR­145 expression in aged rats.

Sevoflurane can produce toxicity to the hippocampal tissues of brain, leading to nerve damage, causing learning and cognitive dysfunction. CircRNAs have been indicated to act as a key mediator in anesthetic neurotoxicity. This study focused on the effect of circ_0016760 on sevoflurane­induced neurological impairment. The GEO database (GSE147277) and RT­qPCR were used to predict and  measure the circ_0016760 expression. The interaction of circ_0016760 and miR­145 was verified by dual­luciferase reporter assay. The CCK­8 assay, flow cytometry, ELISA, ROS kit, MWM test were carried out to measure the cell viability, apoptosis, inflammation indicators, ROS level, and cognitive and memory function of the rats. Sevoflurane exacerbated neurotoxicity by restraining cell viability, inducing cell apoptosis, neuroinflammation, and ROS generation, and causing learning and cognitive dysfunction. Circ_0016760 expression was increased in POCD patients from the GEO database and upregulated after sevoflurane exposure. miR­145 was a target miRNA of circ_0016760. Silencing of circ_0016760 weakened the effect of sevoflurane on cell viability, cell apoptosis, inflammation­related factors, oxidative stress, which could be reversed by miR­145 inhibitor. The animal experiments results showed that circ_0016760 played a protective effect on regulating the cognitive behavior of sevoflurane­treated aged rats, expression of inflammation cytokine, and oxidative stress factors through targeting miR­145 in sevoflurane­treated aged rat's hippocampal neurons. Our results revealed that silencing of circ_0016760 attenuated sevoflurane­induced hippocampal neuron injury by regulating miR­145 expression, which may provide potential insights into the treatment of sevoflurane­induced neurological impairment.

Antigallbladder Carcinoma Activity Analysis of a New Nanometer Drug Delivery System Based on Data Acquisition.

In order to solve the problem of the application of data acquisition in the new nanometer drug delivery system of microscope, a research of antigallbladder carcinoma activity analysis was proposed. Gallbladder carcinoma (GBC) is a common malignant tumor in biliary tract diseases. Due to the lack of specific clinical manifestations in the early stage, GBC has the shortcomings of the hidden onset, the difficult diagnosis, and the high misdiagnosis rate. GBC ranks in the top position among the most common tumors of the digestive system worldwide. The preoperative diagnosis rate is low, and the incidence of accidental gallbladder carcinoma is gradually increasing. Domestic gallbladder carcinoma related to cholecystectomy is not sensitive to radiotherapy and chemotherapy. Surgical resection is still the only effective method for the treatment of accidental gallbladder carcinoma.

[Simulation analysis of the radial growth characteristics of Pinus sylvestris var. mongolica in Hulunbuir Sandy Land by Vaganov-Shashkin Model]. / 呼伦贝尔沙地樟子松径向生长特征的VS模型模拟分析.

The Vaganov-Shashkin model was used to simulate the standardized ring-width chrono-logy of Pinus sylvestris var. mongolica on the sandy land in Hulunbuir. The results showed that the fitting degree was better in the period before 2000 than that after 2000. Thus, the simulation results before 2000 were selected to analyze the radial growth. The results showed that the main growing season of P. sylvestris var. mongolica in Hulunbuir Sandy Land was from May to September each year. Temperature had a significant impact on the early and late growth of P. sylvestris var. mongo-lica. In the prosperous period of the growing season, insufficient soil moisture was the main factor restricting the radial growth. The radial growth rates of P. sylvestris var. mongolica in extremely narrow years were more limited by soil moisture than that in extremely wide years. The radial growth rates in the middle of the growing season (from July to August) showed a decreasing trend in wide and narrow years, indicating that the growth of P. sylvestris var. mongolica was affected by drought stress. Our results were consistent with the characteristics of tree-ring physiological models in semi-arid areas of China. The model was applicable to the radial growth simulation of P. sylvestris var. mongolica in Hulunbuir Sandy Land.

Effects of oxymatrine on the proliferation of human liver cancer Bel-7404 cells: A protocol of systematic review and meta-analysis.

BACKGROUND: This study will examine the effects of oxymatrine on the proliferation of human liver cancer Bel-7404 cells (HLCBC). METHODS: This study will search electronic bibliographic databases available in PUBMED, EMBASE, Cochrane Library, Scopus, Cumulative Index to Nursing and Allied Health Literature, China Biology Medicine, and China National Knowledge Infrastructure. We attempt to search case-controlled studies (CCSs) or randomized controlled studies (RCSs) pertaining to HLCBC from their inception to the February 29, 2020 without limitations of language and publication time. We will include any CCSs or RCSs on exploring oxymatrine on the proliferation of HLCBC. We will assess the methodological quality of CCSs by Newcastle-Ottawa Scale, and RCSs by Cochrane risk of bias tool. Review Manager 5.3 software will be utilized for statistical analysis. RESULTS: The current study will summarize most recent eligible studies to investigate the effects of oxymatrine on the proliferation of HLCBC. CONCLUSION: Its results may provide reliable scientific evidence on effects of oxymatrine on the proliferation of HLCBC. SYSTEMATIC REVIEW REGISTRATION: INPLASY202040026.

Effects of artemisinin on proliferation and apoptosis of human liver cancer HepG2 cells: A protocol of systematic review and meta-analysis.

BACKGROUND: This study will examine the effects of artemisinin on proliferation and apoptosis of human liver cancer HepG2 cells (HLCHG-2C). METHODS: This study will systematically retrieve potential literatures in MEDLINE, Scopus, Web of Science, Cochrane Library, EMBASE, WANGFANG, and China National Knowledge Infrastructure from their initiation to the February 29, 2020. There are not limitations related to the language and publication time. All case-controlled studies (CCSs) or randomized controlled studies (RCSs) will be included in this study which investigated the effects of artemisinin on proliferation and apoptosis of HLCHG-2C. Two independent investigators will examine searched records, collect data from included studies, and will identify their methodological quality. Any divergences will be disentangled by discussion with another investigator. RevMan 5.3 software will be placed to pool the data and to carry out data analysis. RESULTS: This study will summarize all eligible studies to test the effects of artemisinin on proliferation and apoptosis of HLCHG-2C. CONCLUSION: The results of this study will exert evidence to examine the effects of artemisinin on proliferation and apoptosis of HLCHG-2C, and it may benefit further research, patients, and healthcare providers. SYSTEMATIC REVIEW REGISTRATION: INPLASY202040075.

The association between Ki-67 expression and the clinical pathological characteristics of colorectal cancer: A protocol for a systematic review and meta-analysis.

BACKGROUND: This study will explore the association between Ki-67 expression and clinical pathological characteristics (CPC) of colorectal cancer (CC). METHODS: We will search relevant studies from electronic databases (Cochrane Library, PUBMED, EMBASE, Scopus, Cumulative Index to Nursing and Allied Health Literature, China Biology Medicine, and China National Knowledge Infrastructure) from beginning to April 1, 2020 without language and publication time limitations. We will consider all case-controlled studies (CCSs) or randomized controlled studies (RCSs) investigating the association between Ki-67 expression and CPC of CC. We will appraise study quality of CCSs by Newcastle-Ottawa Scale, and RCSs by Cochrane risk of bias tool. Statistical analysis will be carried out by Review Manager 5.3 software. RESULTS: The present study will explore the association between Ki-67 expression and CPC of CC. CONCLUSION: Its findings may summarize scientific evidence of the association between Ki-67 expression and CPC of CC, and may provide helpful evidence for clinical practice.Systematic review registration: PROSPERO CRD42020173795.

Clinicopathological significance of Ki67 expression in colorectal cancer: A protocol of systematic review and meta-analysis.

BACKGROUND: This study will investigate the diagnostic accuracy of Ki67 expression in colorectal cancer (CC). METHODS: A comprehensive search in electronic bibliographic databases (MEDLINE, EMBASE, Cochrane Library, Web of Science, Chinese Biomedical Literature Database, and China National Knowledge Infrastructure) will be performed from inception to the February 29, 2020 with no restrictions to the language and publication status. Two authors will examine the collected studies, extract essential data, and appraise study quality separately. If possible, we will estimate receiver operating characteristic (ROC), sensitivity and specificity by utilizing bivariate random effects and hierarchical summary ROC models. RESULTS: This study will summarize present evidence to explore the diagnostic accuracy of Ki67 expression in CC. CONCLUSION: The findings of this study will clarify the diagnostic accuracy of Ki67 expression in CC. SYSTEMATIC REVIEW REGISTRATION: INPLASY202030009.

The potential role of juvenile hormone acid methyltransferase in methyl farnesoate (MF) biosynthesis in the swimming crab, Portunus trituberculatus.

Juvenile hormone (JH) and methyl farnesoate (MF) play essential roles in the development and reproduction of insects and crustaceans respectively. Juvenile hormone acid methyltransferase (JHAMT) catalyzes the methyl esterification in insect JH biosynthesis, while the corresponding step in crustacean MF biosynthesis was long thought to be catalyzed by farnesoic acid O-methyltransferase (FAMeT). However, the new discovery of JHAMT orthologs in crustaceans indicates that JHAMT may also play essential role in the MF biosynthesis in crustaceans. Here we cloned and characterized the full-length cDNA encoding JHAMT in the swimming crab Portunus trituberculatus (PtJHAMT). Sequence and structure analysis of PtJHAMT revealed that it was composed of a 6-stranded ß sheet with 9 α helices, and contained a signature Sadenosyl-L-methionine (SAM) binding motif, which is the hallmark in all SAM dependent methyltransferases (SAM-MTs). Several active sites that are critical for the interaction of SAM and JH/FA substrate were also conserved in PtJHAMT. The gene expression of PtJHAMT was highly specific to the mandibular organ, which is the sole site of MF synthesis. PtJHAMT expression significantly increased in the late-vitellogenic stage and mature stage, which suggests a possible role of PtJHAMT in modulating ovarian development. The role of PtJHAMT and PtFAMeT in MF biosynthesis was further investigated by RNA interfering (RNAi). Injection of PtJHAMT and PtFAMeT dsRNA both led to a decrease in hemolymph MF titers. Injection of PtHMGR dsRNA caused the decrease in PtJHAMT expression, but had no effect on mRNA level of PtFAMeT. Together these results suggested that JHAMT and FAMeT are both involved in the MF biosynthesis in crustaceans, while the JHAMT is highly specific to FA substrate, and FAMeT may have more catalytic functions.