UCHL-3 as a potential biomarker of ovarian cancer.

OBJECTIVE: This study explored promising prognostic and immune therapeutic candidate biomarkers for OC and determined the expression, prognostic value, and immune effects of UCHL3. METHODS: UCHL3 expression and clinical data were investigated using bioinformatic analysis. CCK8 and transwell assays were conducted to evaluate the impact of UCHL3 on proliferation and migration, and the effects of UCHL3 were further validated in a mouse model. Univariate and least absolute shrinkage and selection operator regression analyses were performed to construct a novel UCHL3-related prognostic risk model. Gene set enrichment analysis (GSEA) and immune analysis were performed to identify the significantly involved functions of UCHL3. Finally, bioinformatic analysis and immunohistochemistry were performed to explore the effect of UCHL3 on chemotherapy. RESULTS: UCHL3 expression was upregulated and associated with worse overall survival (OS) in OC. UCHL3 depletion repressed cell proliferation and migration both in vitro and in vivo. Furthermore, 237 genes were differentially expressed between the high and low UCHL3 expression groups. Subsequently, a UCHL3-related prognostic signature was built based on six prognostic genes (PI3, TFAP2B, MUC7, PSMA2, PIK3C2G, and NME1). Independent prognostic analysis suggested that age, tumor mutational burden, and RiskScore can be used as independent prognostic factors. The immune infiltration analysis and GSEA suggested that UCHL3 expression was related to the immune response. In addition, UCHL3 expression was higher in platinum-resistant OC patients than in platinum-sensitive patients. UCHL3 overexpression was associated with poorer OS. CONCLUSION: UCHL3 overexpression contributes to aggressive progression, poor survival, and chemoresistance in OC. Therefore, UCHL3 may be a candidate prognostic biomarker and potential target for controlling progression and platinum resistance in OC.

Nurses in China lack knowledge of inhaler devices: A cross-sectional study.

Objective: To understand the level of knowledge about inhaler devices among medical staff. Methods: This study evaluated the knowledge of inhalation therapy and the use of inhaler devices among nurses in China. We administered a new self-designed online questionnaire to 1,831 nurses. The questionnaire comprised 11 questions, including the storage location of inhaler devices, steps involved in using inhaler devices, and common errors when using various devices. Results: Among the 1,831 participants, 816(44.57%), 122(6.66%), and 893(48.77%) nurses worked in community, secondary, and tertiary hospitals, respectively. Adequate knowledge of inhaler devices was demonstrated by 20.10%, 8.20%, and 13.10% of nurses working in community, secondary, and tertiary hospitals, respectively. Of the nurses working in community hospitals, 27.70% knew the key points for using inhalers compared to 15.57% in secondary hospitals and 23.18% in tertiary hospitals (p < 0.01). Only 9.50%-26.00% of participants chose correct answers to the 9 questions about the use of inhalers. The accuracy rate of the responses was generally low, and the highest accuracy rate was 26.00%. Conclusion: Knowledge of inhalation therapy was better among nurses working in community hospitals than among those working in high-level hospitals. This is because of the clearer division of work and higher workload in high-level hospitals. Overall, nurses' knowledge of inhalation therapy is low. Furthermore, knowledge about inhaler devices should be strengthened among nurses in Chinese hospitals. It is necessary to create training opportunities for nurses in China to increase their awareness and knowledge regarding the management of chronic respiratory diseases.

Sex Detection of Chicks Based on Audio Technology and Deep Learning Methods.

The sex detection of chicks is an important work in poultry breeding. Separating chicks of different sexes early can effectively improve production efficiency and commercial benefits. In this paper, based on the difference in calls among one-day-old chicks of different sexes, a sex detection method based on chick calls is designed. Deep learning methods were used to classify the calls of chicks and detect their sex. This experiment studies three different varieties of chicks. The short-time zero-crossing rate was used to automatically detect the endpoints of chick calls in audio. Three kinds of audio features were compared: Spectrogram, Cepstrogram and MFCC+Logfbank. The features were used as the input in neural networks, and there were five kinds of neural networks: CNN, GRU, CRNN, TwoStream and ResNet-50. After the cross-comparison experiment of different varieties of chicks, audio features and neural networks, the ResNet-50 neural network trained with the MFCC+Logfbank audio features of three yellow chick calls had the highest test accuracy of 83% when testing Three-yellow chicks' calls. The GRU neural network trained with the Spectrogram audio features of native chick calls had the highest test accuracy of 76.8% when testing Native chicks' calls. The ResNet-50 neural network trained with Spectrogram audio features of flaxen-yellow chick calls had the highest test accuracy of 66.56%when testing flaxen-yellow chick calls. Multiple calls of each chick were detected, and the majority voting method was used to detect the sex of the chicks. The ResNet-50 neural network trained with the Spectrogram of three yellow chick calls had the highest sex detection accuracy of 95% when detecting the three yellow chicks' sex. The GRU neural network trained with the Spectrogram and cepstrogram of native chick calls and the CRNN network trained with the Spectrogram of native chick calls had the highest sex detection accuracy of 90% when detecting the native chicks' sex. The Twostream neural network trained with MFCC+Logfbank of flaxen-yellow chick calls and the ResNet-50 network trained with the Spectrogram of flaxen-yellow chick calls had the highest sex detection accuracy of 80% when detecting the flaxen-yellow chicks' sex. The results of the cross-comparison experiment show that there is a large diversity between the sex differences in chick calls of different breeds. The method is more applicable to chick sex detection in three yellow chicks and less so in native chicks and flaxen-yellow chicks. Additionally, when detecting the sex of chicks of a similar breed to the training chicks, the method obtained better results, while detecting the sex of chicks of other breeds, the detection accuracy was significantly reduced. This paper provides further perspectives on the sex detection method of chicks based on their calls and help and guidance for future research.

Mobile Apps for Drug-Drug Interaction Checks in Chinese App Stores: Systematic Review and Content Analysis.

BACKGROUND: As a computerized drug-drug interaction (DDI) alert system has not been widely implemented in China, health care providers are relying on mobile health (mHealth) apps as references for checking drug information, including DDIs. OBJECTIVE: The main objective of this study was to evaluate the quality and content of mHealth apps supporting DDI checking in Chinese app stores. METHODS: A systematic review was carried out in November 2020 to identify mHealth apps providing DDI checking in both Chinese iOS and Android platforms. We extracted the apps' general information (including the developer, operating system, costs, release date, size, number of downloads, and average rating), scientific or clinical basis, and accountability, based on a multidimensional framework for evaluation of apps. The quality of mHealth apps was evaluated by using the Mobile App Rating Scale (MARS). Descriptive statistics, including numbers and percentages, were calculated to describe the characteristics of the apps. For each app selected for evaluation, the section-specific MARS scores were calculated by taking the arithmetic mean, while the overall MARS score was described as the arithmetic mean of the section scores. In addition, the Cohen kappa (κ) statistic was used to evaluate the interrater agreement. RESULTS: A total of 7 apps met the selection criteria, and only 3 included citations. The average rating score for Android apps was 3.5, with a minimum of 1.0 and a maximum of 4.9, while the average rating score for iOS apps was 4.7, with a minimum of 4.2 and a maximum of 4.9. The mean MARS score was 3.69 out of 5 (95% CI 3.34-4.04), with the lowest score of 1.96 for Medication Guidelines and the highest score of 4.27 for MCDEX mobile. The greatest variation was observed in the information section, which ranged from 1.41 to 4.60. The functionality section showed the highest mean score of 4.05 (95% CI 3.71-4.40), whereas the engagement section resulted in the lowest average score of 3.16 (95% CI 2.81-3.51). For the information quality section, which was the focus of this analysis, the average score was 3.42, with the MCDEX mobile app having the highest score of 4.6 and the Medication Guidelines app having the lowest score of 1.9. For the overall MARS score, the Cohen interrater κ was 0.354 (95% CI 0.236-0.473), the Fleiss κ was 0.353 (95% CI, 0.234-0.472), and the Krippendorff α was 0.356 (95% CI 0.237-0.475). CONCLUSIONS: This study systematically reviewed the mHealth apps in China with a DDI check feature. The majority of investigated apps demonstrated high quality with accurate and comprehensive information on DDIs. However, a few of the apps that had a massive number of downloads in the Chinese market provided incorrect information. Given these apps might be used by health care providers for checking potential DDIs, this creates a substantial threat to patient safety.

circHIPK3 Acts as Competing Endogenous RNA and Promotes Non-Small-Cell Lung Cancer Progression through the miR-107/BDNF Signaling Pathway.

Circular RNAs (circRNAs) act as a crucial part in many human diseases, particularly in cancers. circRNA HIPK3 (circHIPK3) is a special circRNA that may participate in the oncogenesis of non-small-cell lung cancer (NSCLC), even though its latent regulatory mechanism is not very clear. Here, we studied the roles of circHIPK3 in NSCLC. qRT-PCR assay was applied to study the expression of circHIPK3 in NSCLC. The influence of circHIPK3 on NSCLC was estimated by silencing circHIPK3 and miR-107 mock transfection and brain-derived neurotrophic factor (BDNF) overexpression, and the correlation between circHIPK3, miR-107, and BDNF was evaluated by dual-luciferase reporter assay. The results showed that circHIPK3 expression was upregulated in NSCLC cells. circHIPK3 knockdown inhibited the migration and proliferation of NSCLC cells by promoting the expression of miR-107. circHIPK3 could be used as a miR-107 sponge to promote BDNF cell proliferation. The dual-luciferase reporter assay proved that miR-107 was the target of circHIPK3, and miR-107 had an interaction with the 3'untranslated region of BDNF. miR-107 overexpression inhibited BDNF-mediated NSCLC cell proliferation. These results indicate that circHIPK3 promotes tumor progression through a new circHIPK3/miR-107/BDNF axis, which offers potential markers and medical treatment for NSCLC.

YY1-induced lncRNA ZFPM2-AS1 facilitates cell proliferation and invasion in small cell lung cancer via upregulating of TRAF4.

BACKGROUND: Newly identified lncRNA zinc finger protein, FOG family member 2 antisense RNA 1 (ZFPM2-AS1) is identified as an oncogenic gene. However, the role of ZFPM2-AS1 in small cell lung cancer (SCLC) is poorly comprehended. METHODS: The expression of genes in SCLC tissues and cells was measured by qRT-PCR. Colony formation, EdU, CCK-8, transwell and wound healing as well as in vivo assays revealed the function of ZFPM2-AS1 in SCLC. ChIP, luciferase reporter, RIP and RNA pull down assays demonstrated the binding relation among genes. RESULTS: ZFPM2-AS1 was significantly upregulated in SCLC tissues and cells. ZFPM2-AS1 deficiency attenuated SCLC cell proliferation, invasion and migration. In addition, ZFPM2-AS1 was transcriptionally activated by Yin Yang 1 (YY1) factor. Further, miR-3612 was confirmed as downstream miRNA of ZFPM2-AS1. Moreover, TNF receptor associated factor 4 (TRAF4) was the target gene of miR-3612 in SCLC. ZFPM2-AS1, miR-3612 and TRAF4 jointly constituted a competing endogenous RNA (ceRNA) network in SCLC. Finally, TRAF4 could countervail ZFPM2-AS1 downregulation-mediated function on SCLC cell proliferation and invasion in vitro and tumor growth in vivo. CONCLUSION: Our study elucidated the oncogenic effect of ZFPM2-AS1 in SCLC progression, indicating it may be a therapeutic target for SCLC.

Lycofargesiines A-F, further Lycopodium alkaloids from the club moss Huperzia fargesii.

Six undescribed Lycopodium alkaloids (LAs) comprising four lycodine-type (lycofargesiines A-D), one lycopodine-type (lycofargesiine E), and a phlegmarine-type (lycofargesiine F), together with 16 known ones were isolated from the club moss Huperzia fargesii. Their structures and absolute configurations were determined by extensive spectroscopic methods, electronic circular dichroism (ECD) analysis, and density functional theory (DFT) calculations. (7S,8R,12R,13R)-Lycofargesiine A is a rare naturally occurring LA possessing an exocyclic double bond between C-15 and C-16, with ring A being a rare 2,3-dihyropyridone motif. Lycofargesiine D is an uncommon lycodine-type alkaloid featuring a unique N-acetylated tetrahydropyridinyl segment (ring A), whereas lycofargesiine F is the first phlegmarane-type LA bearing two nitrone moieties. In addition to the isolated huperzine A in this study, another two isolates (lycofargesiine C and 16-hydroxyhuperzine A) were also found to show inhibitory activities against acetylcholinesterase (AChE), with IC50 values of 8.63 and 5.18 µM, respectively.

Epigenetics in ovarian cancer: premise, properties, and perspectives.

Malignant ovarian tumors bear the highest mortality rate among all gynecological cancers. Both late tumor diagnosis and tolerance to available chemical therapy increase patient mortality. Therefore, it is both urgent and important to identify biomarkers facilitating early identification and novel agents preventing recurrence. Accumulating evidence demonstrates that epigenetic aberrations (particularly histone modifications) are crucial in tumor initiation and development. Histone acetylation and methylation are respectively regulated by acetyltransferases-deacetylases and methyltransferases-demethylases, both of which are implicated in ovarian cancer pathogenesis. In this review, we summarize the most recent discoveries pertaining to ovarian cancer development arising from the imbalance of histone acetylation and methylation, and provide insight into novel therapeutic interventions for the treatment of ovarian carcinoma.

The RNA binding protein SORBS2 suppresses metastatic colonization of ovarian cancer by stabilizing tumor-suppressive immunomodulatory transcripts.

BACKGROUND: Ovarian cancer constitutes one of the most lethal gynecologic malignancies for females. Currently, early detection strategies and therapeutic options for ovarian cancer are far from satisfactory, leading to high diagnosis rates at late stages and disease relapses. New avenues of therapy are needed that target key processes in ovarian cancer progression. While a variety of non-coding RNAs have been proven to regulate ovarian cancer metastatic progression, the functional roles of RNA-binding proteins (RBPs) in this process are less well defined. RESULTS: In this study, we identify that the RBP sorbin and SH3 domain containing 2 (SORBS2) is a potent suppressor of ovarian cancer metastatic colonization. Mechanistic studies show that SORBS2 binds the 3' untranslated regions (UTRs) of WFDC1 (WAP four-disulfide core domain 1) and IL-17D (Interleukin-17D), two secreted molecules that are shown to act as metastasis suppressors. Enhanced expression of either WFDC1 or IL-17D potently represses SORBS2 depletion-mediated cancer metastasis promotion. By enhancing the stability of these gene transcripts, SORBS2 suppresses ovarian cancer invasiveness and affects monocyte to myeloid-derived suppressor cell and M2-like macrophage polarization, eliciting a tumor-suppressive immune microenvironment. CONCLUSIONS: Our data illustrate a novel post-transcriptional network that links cancer progression and immunomodulation within the tumor microenvironment through SORBS2-mediated transcript stabilization.

Pharmacological activation of estrogen receptor beta augments innate immunity to suppress cancer metastasis.

Metastases constitute the greatest causes of deaths from cancer. However, no effective therapeutic options currently exist for cancer patients with metastasis. Estrogen receptor ß (ERß), as a member of the nuclear receptor superfamily, shows potent tumor-suppressive activities in many cancers. To investigate whether modulation of ERß could serve as a therapeutic strategy for cancer metastasis, we examined whether the selective ERß agonist LY500307 could suppress lung metastasis of triple-negative breast cancer (TNBC) and melanoma. Mechanistically, while we observed that LY500307 potently induced cell death of cancer cells metastasized to lung in vivo, it does not mediate apoptosis of cancer cells in vitro, indicating that the cell death-inducing effects of LY500307 might be mediated by the tumor microenvironment. Pathological examination combined with flow cytometry assays indicated that LY500307 treatment induced significant infiltration of neutrophils in the metastatic niche. Functional experiments demonstrated that LY500307-treated cancer cells show chemotactic effects for neutrophils and that in vivo neutrophil depletion by Ly6G antibody administration could reverse the effects of LY500307-mediated metastasis suppression. RNA sequencing analysis showed that LY500307 could induce up-regulation of IL-1ß in TNBC and melanoma cells, which further triggered antitumor neutrophil chemotaxis. However, the therapeutic effects of LY500307 treatment for suppression of lung metastasis was attenuated in IL1B-/- murine models, due to failure to induce antitumor neutrophil infiltration in the metastatic niche. Collectively, our study demonstrated that pharmacological activation of ERß could augment innate immunity to suppress cancer metastatic colonization to lung, thus providing alternative therapeutic options for cancer patients with metastasis.

Antifungal activity of osthol in vitro and enhancement in vivo through Eudragit S100 nanocarriers.

In vitro interaction of osthol (Ost) and fluconazole (FLC) was investigated against 11 fluconazole-resistant clinical isolates of Candida albicans. Synergistic activities were determined using the checkerboard microdilution assay. The results of agar diffusion test confirmed the synergistic interaction. We used an enteric material Eudragit S100 for preparation of Ost nanoparticle (Ost-NP) to improve the oral bioavailability, biological activity of Ost. The physicochemical characteristics of Ost-S100-NP revealed Ost-S100-NP with mean particle size of 55.4±0.4 nm, encapsulation efficiency of 98.95±0.06%, drug loading efficiency of 23.89±0.25%, yield of 98.5±0.1% and a polydispersity index (PDI) of 0.165. As the Ost concentration-time curve showed, Ost-S100-NP can increase the plasma concentration and relative bioavailability of Ost compared with Ost-suspension by oral administration. In vivo, Ost-S100-NP enhanced the therapeutic efficacy of Ost against FLC-resistant C. albicans in immunosuppressed candidiasis mice model. The available information strongly suggests that Ost-S100-NP may be used as a promising compound against drug-resistant fungi.

Kaempferol induces autophagic cell death of hepatocellular carcinoma cells via activating AMPK signaling.

In the present study, we demonstrate that Kaempferol inhibited survival and proliferation of established human hepatocellular carcinoma (HCC) cell lines (HepG2, Huh-7, BEL7402, and SMMC) and primary human HCC cells. Kaempferol treatment in HCC cells induced profound AMP-activated protein kinase (AMPK) activation, which led to Ulk1 phosphorylation, mTOR complex 1 inhibition and cell autophagy. Autophagy induction was reflected by Beclin-1/autophagy gene 5 upregulation and p62 degradation as well as light chain 3B (LC3B)-I to LC3B-II conversion and LC3B puncta formation. Inhibition of AMPK, via AMPKα1 shRNA or dominant negative mutation, reversed above signaling changes. AMPK inhibition also largely inhibited Kaempferol-induced cytotoxicity in HCC cells. Autophagy inhibition, by 3-methyaldenine or Beclin-1 shRNA, also protected HCC cells from Kaempferol. Kaempferol downregulated melanoma antigen 6, the AMPK ubiquitin ligase, causing AMPKα1 stabilization and accumulation. We conclude that Kaempferol inhibits human HCC cells via activating AMPK signaling.

Long Noncoding RNA LINC00092 Acts in Cancer-Associated Fibroblasts to Drive Glycolysis and Progression of Ovarian Cancer.

The majority of patients with epithelial ovarian cancer are diagnosed at a late stage when the peritoneal metastases exist; however, there is little knowledge of the metastatic process in this disease setting. In this study, we report the identification of the long noncoding RNA LINC00092 as a nodal driver of metastatic progression mediated by cancer-associated fibroblasts (CAF). Prometastatic properties of CAFs in vitro and in vivo were found to associate with elevated expression of the chemokine CXCL14. In clinical specimens, elevated levels of CXCL14 in CAFs also correlated with poor prognosis. Notably, CXCL14-high CAFs mediated upregulation of LINC00092 in ovarian cancer cells, the levels of which also correlated with poor prognosis in patients. Mechanistic studies showed that LINC00092 bound a glycolytic enzyme, the fructose-2,6-biphosphatase PFKFB2, thereby promoting metastasis by altering glycolysis and sustaining the local supportive function of CAFs. Overall, our study uncovered a positive feedback loop in the metabolism of CXCL14-positive CAFs and ovarian cancer cells that is critical for metastatic progression. Cancer Res; 77(6); 1369-82. ©2017 AACR.

LncRNAs: the bridge linking RNA and colorectal cancer.

Long noncoding RNAs (lncRNAs) are transcribed by genomic regions (exceeding 200 nucleotides in length) that do not encode proteins. While the exquisite regulation of lncRNA transcription can provide signals of malignant transformation, lncRNAs control pleiotropic cancer phenotypes through interactions with other cellular molecules including DNA, protein, and RNA. Recent studies have demonstrated that dysregulation of lncRNAs is influential in proliferation, angiogenesis, metastasis, invasion, apoptosis, stemness, and genome instability in colorectal cancer (CRC), with consequent clinical implications. In this review, we explicate the roles of different lncRNAs in CRC, and the potential implications for their clinical application.

Acute temperature and cadmium stress response characterization of small heat shock protein 27 in large yellow croaker, Larimichthys crocea.

Small heat shock proteins (sHSPs) are a group of molecular chaperones and play a crucial role in cell response to various stresses. In this study, we cloned and sequenced a small heat shock protein 27 (LcHSP27) cDNA from large yellow croaker, Larimichthys crocea. The full-length cDNA of LcHSP27 was 1227 bp, including a 5'-terminal untranslated region (UTR) of 54 bp, a 3'-terminal UTR of 561 bp and an open reading frame (ORF) of 612 bp encoding a polypeptide of 203 amino acids. Three conserved phosphorylation sites of serine were identified in the deduced LcHSP27 amino acid sequence at positions 15, 91 and 95 respectively, and a typical α-crystallin domain was at positions 96-193. Phylogenetic analysis revealed that LcHSP27 was categorized together with the HSP27 obtained from other fish. And a closer phylogenetic relationship of HSP27 was found with HSP22, then HSP30. Quantitative real-time reverse transcription PCR (qRT-PCR) analysis indicated the strongest expression of LcHSP27 in heart. However, expression of LcHSP27 in other examined tissues including muscle, brain, liver, spleen, kidney, gill, and blood was very weak. The impact of temperature and cadmium (Cd(2+)) stress on LcHSP27 expression was tested in liver and brain. The results showed that the levels of LcHSP27 expression increased significantly after low temperature (19°C) and high temperature (27°C and 31°C) stress both in liver and brain. And low temperature stress induced a higher LcHSP27 expression in liver. More important, LcHSP27 expression showed a dramatic up-regulation after a combined stress of temperature and cadmium (p<0.05). These results reveal that HSP27 may play an important role in the large yellow croaker response to temperature and cadmium stress.

Characterization of a RacGTPase up-regulated in the large yellow croaker Pseudosciaena crocea immunity.

The Rac proteins are members of the Rho family of small G proteins and are implicated in the regulation of several pathways, including those leading to cytoskeleton reorganization, gene expression, cell proliferation, cell adhesion and cell migration and survival. In this investigation, a Rac gene (named as LycRac gene) was obtained from the large yellow croaker and it was expressed in Escherichia coli and purified. Subsequently the specific antibody was raised using the purified fusion protein (GST-LycRac). Moreover, the GTP-binding assay showed that the LycRac protein had GTP-binding activity. The LycRac gene was ubiquitously transcribed and expressed in 9 tissues. Quantitative real-time RT-PCR and Western blot analysis revealed the highest expression in gill and the weakest expression in spleen. Time-course analysis revealed that LycRac expression was obviously up-regulated in blood, spleen and liver after immunization with polyinosinic polycytidynic acid (poly I:C), formalin-inactive Gram-negative bacterium Vibrio parahemolyticus and bacterial lipopolysaccharides (LPS). These results suggested that LycRac protein might play an important role in the immune response against microorganisms in large yellow croaker.

Oxidative stress potentiated by diallylsulfide, a selective CYP2E1 inhibitor, in isoniazid toxic effect on rat primary hepatocytes.

Isoniazid (INH), one of the first-line antituberculosis drugs, has potential liver toxicity. Mechanisms reported by previous studies mainly focused on oxidative stress. In the present study, we investigated acute effects of diallylsulfide (DAS), a selective CYP2E1 inhibitor, on reduced glutathione (GSH) and reactive oxygen species (ROS) levels in rat primary hepatocytes treated with INH. In cultures treated with INH for 1, 4, 8h, significant loss of GSH content and decrease of ROS levels were observed. Moreover, when hepatocytes were co-treated with INH and 1mM DAS, accelerated GSH depletion and increased ROS production appeared. Further more, rat primary hepatocytes survival rates decreased significantly in cultures treated with INH together with DAS than in cultures treated with INH alone after 24h. In conclusion, DAS could potentiate INH toxic effect and this is the first study reporting the effect of DAS on oxidative stress in INH-induced hepatocytotoxicity.