Identification of triciribine as a novel myeloid cell differentiation inducer

by Souma Suzuki, Susumu Suzuki, Yuri Sato-Nagaoka, Chisaki Ito, Shinichiro Takahashi

Differentiation therapy using all-trans retinoic acid (ATRA) for acute promyelocytic leukemia (APL) is well established. However, because the narrow application and tolerance development of ATRA need to be improved, we searched for another efficient myeloid differentiation inducer. Kinase activation is involved in leukemia biology and differentiation block. To identify novel myeloid differentiation inducers, we used a Kinase Inhibitor Screening Library. Using a nitroblue tetrazolium dye reduction assay and real-time quantitative PCR using NB4 APL cells, we revealed that, PD169316, SB203580, SB202190 (p38 MAPK inhibitor), and triciribine (TCN) (Akt inhibitor) potently increased the expression of CD11b. We focused on TCN because it was reported to be well tolerated by patients with advanced hematological malignancies. Nuclear/cytoplasmic (N/C) ratio was significantly decreased, and myelomonocytic markers (CD11b and CD11c) were potently induced by TCN in both NB4 and acute myeloid leukemia (AML) M2 derived HL-60 cells. Western blot analysis using NB4 cells demonstrated that TCN promoted ERK1/2 phosphorylation, whereas p38 MAPK phosphorylation was not affected, suggesting that activation of the ERK pathway is involved in TCN-induced differentiation. We further examined that whether ATRA may affect phosphorylation of ERK and p38, and found that there was no obvious effect, suggesting that ATRA induced differentiation is different from TCN effect. To reveal the molecular mechanisms involved in TCN-induced differentiation, we performed microarray analysis. Pathway analysis using DAVID software indicated that “hematopoietic cell lineage” and “cytokine-cytokine receptor interaction” pathways were enriched with high significance. Real-time PCR analysis demonstrated that components of these pathways including IL1β, CD3D, IL5RA, ITGA6, CD44, ITGA2B, CD37, CD9, CSF2RA, and IL3RA, were upregulated by TCN-induced differentiation. Collectively, we identified TCN as a novel myeloid cell differentiation inducer, and trials of TCN for APL and non-APL leukemia are worthy of exploration in the future.

Intolerance of uncertainty and mental health in China “Post-pandemic” age: The mediating role of difficulties in emotion regulation

by Zi-Hao Gao, Jun Li

The Chinese government adjusted its national epidemic prevention and control policy in December 2022 after the worldwide declaration of COVID-19 as a common influenza. After the policy adjustment, there has been widespread infection in China, which has brought a lot of uncertainty to the lives and studies of Chinese university students. This study focused on the impact of the intolerance of uncertainty for COVID-19 (IUC) on the emotional and mental health of college students in China “Post-pandemic” age. This study examined the mediating role of difficulties in emotion regulation (DER) between IUC and mental health (MH). 1,281 university students in China were surveyed using the intolerance of uncertainty for COVID-19 scale, the difficulties in emotion regulation scale and the mental health scale. A structural equation model was used to test the hypothesis model, and it was shown that IUC had a significant negative effect on the MH of college students and a significant positive effect on the DER. DER had a significant negative effect on the MH, and DER had a complete mediation effect between IUC and MH. The findings of this study enrich our understanding of the influencing factors of mental health of university students under the background of post-epidemic in China, and provide practical reference for universities on how to prevent mental health problems under the current uncertain environment in China.

Authenticity: Effective emotional labor strategies on teaching efficacy of university teachers in China

by Jiuhua Zheng, Yuqing Geng, Juan Gao, Qinjun Xiang

Based on the job demands-resources model, this study explored the relationships among emotional labor strategies, job demands of teaching, job resources, and teaching efficacy for university teachers. The results, based on a sample of 316 university teachers from China, showed that the teachers preferred to convey authenticity by expressing deep acting and naturally felt emotion. Furthermore, deep acting and naturally felt emotions were found to have a significant positive effect on teaching efficacy, whereas surface acting did not have any significant relationship with teaching efficacy. As organizational variables, job demands significantly positively affected surface acting, deep acting, and naturally felt emotion. In contrast, job resources positively affected surface and deep acting, but no significant relationship with naturally felt emotions was found. These results highlight that university teachers from China could benefit from adopting authentic emotional labor strategies, specifically deep acting and naturally felt emotions, as the most effective strategies in teaching. Based on the above findings, this paper concludes with recommendations for university administrators to alleviate the emotional labor of college faculty. For example, universities should pay more attention to teachers’ emotional state, provide resources to support them, and give more space and support to college teachers in teaching.

Safety and efficacy of bempedoic acid among patients with statin intolerance and those without: A meta-analysis and a systematic randomized controlled trial review

by Yi Li, Hongyu Gao, Jinghui Zhao, Liqing Ma, Dan Hu

Objective

Bempedoic acid, an innovative oral medication, has garnered significant interest in recent times due to its potential as a therapeutic intervention for hypercholesterolemia. Nonetheless, the outcomes of the initial investigations might have been more definitive and coherent. Our objective was to perform a quantitative meta-analysis in order to evaluate bempedoic acid’s safety and effectiveness.

Methods

A search was conducted on ClinicalTrials.gov, and PubMed from the time of inception until September 28, 2023. Randomized controlled trials comparing the safety and efficacy of bempedoic acid among patients with statin intolerance and those without were included in our analysis. The trial outcomes were summarized using a random effects model and were provided as mean differences or odds ratios (ORs) with a confidence interval of 95%. Additionally, trial heterogeneity and the possibility of bias were evaluated and investigated.

Results

Bempedoic acid treatment reduced low-density lipoprotein cholesterol levels more than placebo (mean difference -2.97%, 95% CI -5.89% to -0.05%), according to a pooled analysis of 16 eligible trials. The risk of death (OR 1.18, 95% CI 0.70 to 1.98) and muscle-associated occurrences (OR 1.00, 95% CI 0.77 to 1.31) was not impacted by bempedoic acid. In contrast, discontinuation of treatment was more frequently caused by adverse events in the bempedoic acid group (OR 1.13, 95% CI 1.01 to 1.27).

Conclusions

In patients with statin intolerance as well as those without, bempedoic acid is a safe and efficacious lipid-lowering agent, according to findings from randomized controlled trials.

Exploring the resistance mechanism of triple-negative breast cancer to paclitaxel through the scRNA-seq analysis

by Wei Gao, Linlin Sun, Jinwei Gai, Yinan Cao, Shuqun Zhang

Background

The triple negative breast cancer (TNBC) is the most malignant subtype of breast cancer with high aggressiveness. Although paclitaxel-based chemotherapy scenario present the mainstay in TNBC treatment, paclitaxel resistance is still a striking obstacle for cancer cure. So it is imperative to probe new therapeutic targets through illustrating the mechanisms underlying paclitaxel chemoresistance.

Methods

The Single cell RNA sequencing (scRNA-seq) data of TNBC cells treated with paclitaxel at different points were downloaded from the Gene Expression Omnibus (GEO) database. The Seurat R package was used to filter and integrate the scRNA-seq expression matrix. Cells were further clustered by the FindClusters function, and the gene marker of each subset was defined by FindAllMarkers function. Then, the hallmark score of each cell was calculated by AUCell R package, the biological function of the highly expressed interest genes was analyzed by the DAVID database. Subsequently, we performed pseudotime analysis to explore the change patterns of drug resistance genes and SCENIC analysis to identify the key transcription factors (TFs). Finally, the inhibitors of which were also analyzed by the CTD database.

Results

We finally obtained 6 cell subsets from 2798 cells, which were marked as AKR1C3+, WNT7A+, FAM72B+, RERG+, IDO1+ and HEY1+HCC1143 cell subsets, among which the AKR1C3+, IDO1+ and HEY1+ cell subsets proportions increased with increasing treatment time, and then were regarded as paclitaxel resistance subsets. Hallmark score and pseudotime analysis showed that these paclitaxel resistance subsets were associated with the inflammatory response, virus and interferon response activation. In addition, the gene regulatory networks (GRNs) indicated that 3 key TFs (STAT1, CEBPB and IRF7) played vital role in promoting resistance development, and five common inhibitors targeted these TFs as potential combination therapies of paclitaxel were identified.

Conclusion

In this study, we identified 3 paclitaxel resistance relevant IFs and their inhibitors, which offers essential molecular basis for paclitaxel resistance and beneficial guidance for the combination of paclitaxel in clinical TNBC therapy.

PIK3CA regulates development of diabetes retinopathy through the PI3K/Akt/mTOR pathway

by Ruijuan Guan, Zefeng Kang, Ling Li, Xin Yan, Tianpeng Gao

Objective

To explore their association with the development of diabetes retinopathy (DR), single nucleotide polymorphism (SNP) mutations were screened out by high-throughput sequencing and validated in patients diagnosed with DR. To understand the role of PIK3CA in the pathogenesis of DR and explore the relationship between PIK3CA,phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR),and DR, the effect of PIK3CA.rs17849079 mutation was investigated in a DR cell model.

Methods

Twelve patients diagnosed with DR at the Qinghai Provincial People’s Hospital from September 2020 to June 2021 were randomly selected as the case group, while 12 healthy subjects of similar age and gender who underwent physical examination in Qinghai Provincial People’s Hospital physical examination center during the same period were randomly selected as the control group. Blood samples (2 mL) were collected from both groups using EDTA anticoagulant blood collection vessels and frozen at −20°C for future analysis. SNP mutations were detected by high-throughput sequencing, and the shortlisted candidates were subjected by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. The detected SNP candidates were verified by expanding the sample size (first validation: 56 patients in the case group and 58 controls; second validation: 157 patients in the case group and 96 controls). A lentivirus vector carrying mutated or wild-type PIK3CA.rs17849079 was constructed. ARPE-19 cells were cultured in a medium supplemented with 10% fetal bovine serum (FBS) to establish a DR cell model. PIRES2-PIK3CA-MT and PIRES2-PIK3CA-WT vectors were transfected into DR model cells, which were categorized into control, mannitol, model, empty vector, PIK3CA wild-type, and PIK3CA mutant-type groups. Cell activity was detected by the cell counting kit (CCK)-8 assay, and cellular apoptosis was evaluated by flow cytometry. Glucose concentration and levels of cytokines tumor necrosis factor (TNF)-α and interleukin (IL)-1β were detected using enzyme-linked immunosorbent assay kits. The expression of PIK3CA, AKT1, mTOR, and VEGF genes was detected by real-time quantitative polymerase chain reaction (RT-qPCR), while the expression of PI3K, p-PI3K, AKT1, p-AKT1, mTOR, p-mTOR, and VEGF proteins was detected by western blotting.

Results

The mutated SNPs were mainly enriched in the PI3K/AKT pathway, calcium ion pathway, and glutamatergic synaptic and cholinergic synaptic signaling pathways. Seven SNPs, including PRKCE.rs1533476, DNAH11.rs10485983, ERAP1.rs149481, KLHL1.rs1318761, APOBEC3C.rs1969643, FYN.rs11963612, and KCTD1.rs7240205, were not related to the development of DR. PIK3CA.rs17849079 was prone to C/T mutation. The risk of DR increased with the presence of the C allele and decreased in the presence of the T allele. High glucose induced the expression of PIK3CA and VEGF mRNAs as well as the expression of PI3K, p-PI3K, p-AKT1, p-mTOR, and VEGF proteins in ARPE-19 cells, which led to secretion of inflammatory factors TNF-αand IL-1, cell apoptosis, and inhibition of cell proliferation. The PIK3CA.rs17849079 C allele accelerated the progression of DR. These biological effects were inhibited when the C allele of PIK3CA.rs17849079 was mutated to T allele.

Conclusion

The mutated SNP sites in patients with DR were mainly enriched in PI3K/AKT, calcium ion, and glutamatergic synaptic and cholinergic synaptic signaling pathways. The rs17849079 allele of PIK3CA is prone to C/T mutation where the C allele increases the risk of DR. High glucose activates the expression of PIK3CA and promotes the phosphorylation of PI3K, which leads to the phosphorylation of AKT and mTOR. These effects consequently increase VEGF expression and accelerate the development of DR. The C to T allele mutation in PIK3CA.rs17849079 can play a protective role and reduce the risk of DR.

Molecular cloning and functional analysis of 4-coumarate: CoA ligases from <i>Marchantia paleacea</i> and their roles in lignin and flavanone biosynthesis

by Shuai Gao, Xin-Yan Liu, Rong Ni, Jie Fu, Hui Tan, Ai-Xia Cheng, Hong-Xiang Lou

Phenylpropanoids play important roles in plant physiology and the enzyme 4-coumarate: coenzyme A ligase (4CL) catalyzes the formation of thioesters. Despite extensive characterization in various plants, the functions of 4CLs in the liverwort Marchantia paleacea remain unknown. Here, four 4CLs from M. paleacea were isolated and functionally analyzed. Heterologous expression in Escherichia coli indicated the presence of different enzymatic activities in the four enzymes. Mp4CL1 and Mp4CL2 were able to convert caffeic, p-coumaric, cinnamic, ferulic, dihydro-p-coumaric, and 5-hydroxyferulic acids to their corresponding CoA esters, while Mp4CL3 and Mp4CL4 catalyzed none. Mp4CL1 transcription was induced when M. paleacea thalli were treated with methyl jasmonate (MeJA). The overexpression of Mp4CL1 increased the levels of lignin in transgenic Arabidopsis. In addition, we reconstructed the flavanone biosynthetic pathway in E. coli. The pathway comprised Mp4CL1, co-expressed with chalcone synthase (CHS) from different plant species, and the efficiency of biosynthesis was optimal when both the 4CL and CHS were obtained from the same species M. paleacea.

Regional anaesthesia practice in public hospitals in Botswana: A cross-sectional study

by Mamo Kassa, Farai Madzimbamuto, Gaone Kediegite, Eugene Tuyishime

Introduction

Little is known about the regional anesthesia practice in low resources settings (LRS). The aim of this study was to describe the regional anesthesia capacity, characteristics of regional anesthesia practice, and challenges and solutions of practicing safe regional anesthesia in public hospitals in Botswana.

Methods

This was a cross-sectional survey of anesthesia providers working in public hospitals in Botswana. A purposive sampling method of public hospitals was used to achieve representation of different hospital levels across Botswana. Paper-based questionnaires were sent to anesthesia providers from selected hospitals. Descriptive statistics were used for analysis.

Results

Questionnaires were distributed to 47 selected anesthesia providers from selected hospitals; 38 (80.9%) were returned. Most participants were nurse anesthetists and medical officers (57.8%). All hospitals perform spinal anesthesia; however, other regional techniques were performed by a small number of participants in one referral hospital. Most hospitals had adequate regional anesthesia drugs and sedation medications, however, most hospitals (except one referral hospital) lacked ultrasound machine and the regional anesthesia kit. The common challenges reported were lack of knowledge and skills, lack of equipment and supplies, and lack of hospital engagement and support. Some solutions were proposed such as regional anesthesia training and engaging the hospital management to get resources.

Conclusions

The results of this study suggest that spinal anesthesia is the most common regional anesthesia technique performed by anesthesia providers working in public hospitals in Botswana followed by few upper limb blocks. However, most public hospitals lack enough training capacity, equipment, and supplies for regional anesthesia. More engagement of the hospital management, investment in regional anesthesia resources, and training are needed in order to improve the regional anesthesia capacity and provide safe surgery and anesthesia in Botswana.

Identification of genes regulated by lipids from seaweed Susabinori (<i>Pyropia yezoensis</i>) involved in the improvement of hepatic steatosis: Insights from RNA-Seq analysis in obese <i>db</i>/<i>db</i> mice

by Sayaka Iizasa, Koji Nagao, Keisuke Tsuge, Yukio Nagano, Teruyoshi Yanagita

Hepatic steatosis is an early stage in the progression of non-alcoholic fatty liver disease (NAFLD) and can lead to the development of non-alcoholic steatohepatitis (NASH), a major cause of liver-related morbidity and mortality. Identification of dietary components that can alleviate hepatic steatosis is crucial for developing effective therapeutic strategies for NAFLD. Recently, we demonstrated the impact of lipids extracted from the marine red alga Susabinori (Pyropia yezoensis) in a murine model of type 2-diabete (db/db). We found that Susabinori lipids (SNL), abundant in eicosapentaenoic acid (EPA)-containing polar lipids, protected against obesity-induced hepatic steatosis in db/db mice. To understand the specific genes or biological pathways underlying the effects of SNL, we conducted RNA-Seq analysis of the hepatic transcriptome. By performing comparative analysis of differentially expressed genes between normal mice and db/db mice consuming a control diet, as well as SNL-fed db/db mice, we identified the 15 SNL-dependent up-regulated genes that were down-regulated in db/db mice but up-regulated by SNL feeding. Gene ontology and pathway analysis on these 15 genes demonstrated a significant association with the metabolisms of arachidonic acid (AA) and linoleic acid (LA). Furthermore, we observed alterations in the expression levels of monoacylglycerol lipase (Magl) and fatty acid-binding protein 4 (Fabp4) in the SNL-fed db/db mice, both of which are implicated in AA and LA metabolism. Additionally, the livers of SNL-fed db/db mice exhibited reduced levels of AA and LA, but a high accumulation of EPA. In conclusion, the SNL diet might affect the metabolisms of AA and LA, which contribute to the improvement of hepatic steatosis. Our findings provide insights into the molecular mechanisms underlying the beneficial effects of SNL.