Multidimensional Optimization of Saccharomyces cerevisiae for Carotenoid Overproduction.

Microbial synthesis of carotenoids is a highly desirable alternative to plant extraction and chemical synthesis. In this study, we investigated multidimensional strategies to improve the carotenoid synthesis in the industrial workhorse of Saccharomyces cerevisiae. First, we rewired the yeast central metabolism by optimizing non-oxidative glycolysis pathway for an improved acetyl-CoA supply. Second, we restricted the consumption of farnesyl pyrophosphate (FPP) by the down-regulation of squalene synthase using the PEST degron. Third, we further explored the human lipid binding/transfer protein saposin B (hSapB)-mediated metabolic sink for an enhanced storage of lipophilic carotenoids. Last, the copper-induced GAL expression system was engineered to function in the yeast-peptone-dextrose medium for an increased biomass accumulation. By combining the abovementioned strategies, the final engineered yeast produced 166.79 ± 10.43 mg/l ß-carotene in shake flasks, which was nearly 5-fold improvement of the parental carotenoid-producing strain. Together, we envision that multidimensional strategies reported here might be applicable to other hosts for the future industrial development of carotenoid synthesis from renewable feedstocks.

HBV infection effects prognosis and activates the immune response in intrahepatic cholangiocarcinoma.

BACKGROUND: The impact of HBV infection on the prognosis of patients with intrahepatic cholangiocarcinoma (ICC) remains uncertain, and the underlying mechanism has not been elucidated. This study aims to explore the potential mechanism via clinical perspectives and immune features. METHODS: We retrospectively reviewed 1308 patients with ICC treated surgically from January 2007 to January 2015. Then, we compared immune-related markers using immunohistochemistry staining to obtain the gene expression profile GSE107943 and related literature for preliminary bioinformatics analysis. Subsequently, we conducted a drug sensitivity assay to validate the role of TNFSF9 in the ICC organoid-autologous immune cell coculture system and in the patient-derived organoids-based xenograft platform. RESULTS: The analysis revealed that tumors in patients without HBV infection exhibited greater size and a higher likelihood of lymphatic metastasis, tumor invasion, and relapse. After resection, HBV-infected patients had longer survival time than uninfected patients (p<0.01). Interestingly, the expression of immune-related markers in HBV-positive patients with ICC was higher than that in uninfected patients (p<0.01). The percentage of CD8+ T cells in HBV-positive tissue was higher than that without HBV infection (p<0.05). We screened 21 differentially expressed genes and investigated the function of TNFSF9 through bioinformatics analyses. The expression of TNFSF9 in ICC organoids with HBV infection was lower than that in organoids without HBV infection. The growth of HBV-negative ICC organoids was significantly inhibited by inhibiting the expression of TNFSF9 with a neutralizing antibody. Additionally, the growth rate was faster in HbsAg (-) ICC patient-derived organoids-based xenograft model than in HbsAg (+) group. CONCLUSIONS: The activation of the immune response induced by HBV infection makes the prognosis of HBV-positive patients with ICC differ from that of uninfected patients.

Presence of onco-fetal neighborhoods in hepatocellular carcinoma is associated with relapse and response to immunotherapy.

Onco-fetal reprogramming of the tumor ecosystem induces fetal developmental signatures in the tumor microenvironment, leading to immunosuppressive features. Here, we employed single-cell RNA sequencing, spatial transcriptomics and bulk RNA sequencing to delineate specific cell subsets involved in hepatocellular carcinoma (HCC) relapse and response to immunotherapy. We identified POSTN+ extracellular matrix cancer-associated fibroblasts (EM CAFs) as a prominent onco-fetal interacting hub, promoting tumor progression. Cell-cell communication and spatial transcriptomics analysis revealed crosstalk and co-localization of onco-fetal cells, including POSTN+ CAFs, FOLR2+ macrophages and PLVAP+ endothelial cells. Further analyses suggest an association between onco-fetal reprogramming and epithelial-mesenchymal transition (EMT), tumor cell proliferation and recruitment of Treg cells, ultimately influencing early relapse and response to immunotherapy. In summary, our study identifies POSTN+ CAFs as part of the HCC onco-fetal niche and highlights its potential influence in EMT, relapse and immunotherapy response, paving the way for the use of onco-fetal signatures for therapeutic stratification.

Genomic and transcriptomic characteristics of 12 novel primary cell lines derived from three patients with cholangiocarcinoma.

Cholangiocarcinoma (CCA) is an aggressive bile duct malignancy with poor prognosis. To improve our understanding of the biological characteristics of CCA and develop effective therapies, appropriate preclinical models are required. Here, we established and characterized 12 novel patient-derived primary cancer cell (PDPC) models using multi-region sampling. At the genomic level of PDPCs, we observed not only commonly mutated genes, such as TP53, JAK3, and KMT2C, consistent with the reports in CCA, but also specific mutation patterns in each cell line. In addition, specific expression patterns with distinct biological functions and pathways involved were also observed in the PDPCs at the transcriptomic level. Furthermore, the drug-sensitivity results revealed that the PDPCs exhibited different responses to the six commonly used compounds. Our findings indicate that the established PDPCs can serve as novel in vitro reliable models to provide a crucial molecular basis for improving the understanding of tumorigenesis and its treatment.

[Effect of Intolerance of Uncertainty on Fear of Childbirth in Primiparas : A Moderated Mediating Model].

Objective: To explore the mediating effect of childbirth self-efficacy on the impact pathway of intolerance of uncertainty on the fear of childbirth in primiparas in the second and third trimesters and the potential moderating effect of perceived partner responsiveness. Methods: A total of 429 primiparas in their second and third trimesters completed the survey, which included general information questionnaire, Intolerance of Uncertainty Scale, Childbirth Self-Efficacy Inventory, Perceived Partner Responsiveness Scale, and Childbirth Attitudes Questionnaire. Rank sum test was used to compare the scores for the fear of childbirth among different groups and Spearman's correlation was used to analyze the scores for all the scales. In addition, the data were centrally processed by using PROCESS V3.4.1 Model 4 (a simple mediation model), Model 5 (the direct path of the mediation model was regulated), and non-parametric Bootstrap method to test the mediation effect and moderation effect. Results: The study showed that 54.31% of the participants experienced fear of childbirth. Their scores for intolerance of uncertainty, childbirth self-efficacy, and perceived partner responsiveness were 15.00 (8.00), 240.00 (75.00), and 72.00 (19.00), respectively. There were significant differences in the scores for the fear of childbirth scale among pregnant women of different age groups, gestational weeks, employment statuses, and average per capita monthly income of the family ( P<0.05). According to our findings, intolerance of uncertainty directly and positively impacted on fear of childbirth ( ß=0.76, P<0.001), with childbirth self-efficacy playing partial mediation role between them, its indirect effect being 0.05 and the contribution rate being 6.17%. In addition, after the scores of Perceived Partner Responsiveness Scale were added to the model, perceived partner responsiveness had no significant predictive effect for fear of childbirth, but the product term of the scores for Perceived Partner Responsiveness Scale and Intolerability Uncertainty Scale had significant predictive effect for fear of childbirth ( ß=0.01, P<0.05), which suggested that perceived partner responsiveness also played a moderating role between intolerance of uncertainty and fear of childbirth. Conclusion: Health care providers can help primiparas reduce fear of childbirth and improve their childbirth experience by reducing perceived intolerance of uncertainty, improving family support, and teaching coping strategies.

Quantitative energy spectrum CT in differential diagnosis of aldosterone-producing adenoma and cortisol-producing adenoma.

Background: Patients with aldosterone-producing adenoma (APA) and cortisol-producing adenoma (CPA) show some similar clinical symptoms, and a large overlap of conventional imaging manifestations, which make the differentiation difficult. The purpose of our study was to explore the value of gemstone spectral imaging (GSI) dual-energy computed tomography (DECT) in differential diagnosis of APA and CPA, screen out meaningful energy spectral indicators and provide theoretical basis for the differential diagnosis of the two. Methods: We retrospectively analyzed the imaging and clinical data of 30 patients with APA and 27 patients with CPA who underwent GSI DECT in The First Affiliated Hospital of Zhengzhou University (a tertiary care institution). Patients were consecutively enrolled in this study, and the quantitative DECT parameters were compared between the APA and CPA groups by two-sample test. The diagnostic efficacies were evaluated by receiver operating characteristic (ROC) analysis. Results: DECT parameters including CT (computed tomography) values at 40-70 keV in the arterial phase, concentrations of I (H2O) and fat (I) in the arterial phase, and the effective atomic number in the venous phase, were significantly different between the APA and CPA groups (all P<0.001), and the area under the curve (AUC) values are 0.80, 0.79, 0.88, 0.76, 0.82, 0.87, and 0.86. Conclusions: DECT quantitative parameters can effectively identify APA and CPA, the CT values at 40 and 60 keV in the arterial phase, the normalized CT value at 60 keV, the I (H2O), fat (I) concentration in the arterial phase and the effective atomic number parameter in the venous phase had valuable diagnostic performance.

Effect of urokinase-type plasminogen activator combined with clinical stage and Barcelona Clinic Liver Cancer stage on the prognosis of patients with hepatocellular carcinoma.

Background: The aim of this investigation is to evaluate the association and potential mechanism between plasminogen activator urokinase (PLAU) and the prognosis of patients with liver hepatocellular carcinoma (LIHC). Methods: We verified PLAU expression and its correlation with LIHC patients' prognosis in The Cancer Genome Atlas (TCGA) database. The interaction network for protein-gene was established in the GeneMania database and the STRING database, and the association between PLAU and immune cells was assessed in Tumor Immune Estimation Resource (TIMER) and TCGA databases. The potential physiological mechanism was elucidated by the Gene Set Enrichment Analysis (GSEA) enrichment assessment. Finally, the individual clinical data of 100 LIHC patients were retrospectively evaluated to further analyze the clinical value of PLAU. Results: The PLAU expression in LIHC tissues was greater than in paracancerous tissues, and LIHC patients with low PLAU expression had better disease-specific survival (DSS), overall survival (OS), and progression free interval (PFI) than those with high PLAU expression. In the TIMER database, the PLAU expression was positively associated with six kinds of infiltrating immune cells: CD4+ T, neutrophils, CD8+ T, macrophages, B, and dendritic cells, while GSEA enrichment analysis indicated PLAU may impact the biological activities of LIHC by taking part in MAPK and JAK_STAT signaling pathways, angiogenesis, and P53. There were statistically significant differences in T-stage and Edmondson grading between the two groups of patients with high and low expression of PLAU (P<0.05). The tumor progression rates were 88% (44/50) and 92% (46/50) respectively in the low and high PLAU groups, with early recurrence rates of 60% (30/50) and 72% (36/50), and median PFS of 29.5 and 23 months, respectively. The COX regression analysis showed PLAU expression and CS and Barcelona Clinic Liver Cancer (BCLC) stages were independent prognostic factors affecting tumor progression in LIHC patients. Conclusions: The decreased expression of PLAU can prolong the DSS, OS, and PFI in LIHC patients, and can be utilized as a novel predictive index. PLAU combined with CS staging and BCLC staging has good clinical value in the early screening and prognosis of LIHC. These results reveal an efficient approach for developing anticancer strategies against LIHC.

Technical comparison of continuous versus intermittent perfusion computed tomography computed tomography scans of the human pancreas.

Background: Pancreatic perfusion computed tomography (CT) imaging is increasingly used for neoplastic grading, predicting prognosis, and evaluating the response to therapy. To optimize the clinical pancreatic CT perfusion imaging methods, we evaluated 2 different CT scanning protocols concerning pancreas perfusion parameters. Methods: A retrospective study was conducted on 40 patients who underwent whole pancreas CT perfusion scanning in The First Affiliated Hospital of Zhengzhou University. Of these 40 patients, 20 patients in group A underwent continuous perfusion scanning, while 20 patients in group B underwent intermittent perfusion scanning. For group A, continuous axial scanning was performed 25 times, and the total scan time was 50 s. For group B, arterial phase helical perfusion scanning was performed 8 times, and then venous phase helical perfusion scanning was performed 15 times, with a total scan time of 64.6 to 70.0 s. A comprehensive list of perfusion parameters between different parts of the pancreas and the 2 groups were compared. The effective radiation dose for the 2 scanning methods was analyzed. Results: The parameter of the mean slope of increase (MSI) at different pancreatic parts in group A differed (P=0.028). The pancreas head had the lowest value, and the tail had the highest (about a 20% difference). In group A compared to group B, the blood volume of the pancreatic head was smaller (15.256±2.925 vs. 16.953±3.602), the positive enhanced integral was smaller (0.307±0.050 vs. 0.344±0.060) and the permeability surface was larger (34.205±9. 612 vs. 24.377±8.413); the blood volume of the pancreatic neck was smaller (13.940±2.691 vs. 17.173±3.918), the positive enhanced integral was smaller (0.304±0.088 vs. 0.361±0.051) and the permeability surface was larger (34.898±11.592 vs. 25. 794±8.149); the blood volume of the pancreatic body was smaller (16.142±4.006 vs. 18.401±2.513), the positive enhanced integral was smaller (0.305±0.093 vs. 0.342±0.048) and the permeability surface was larger (28.861±10.448 vs. 22.158±6. 017); the blood volume of the pancreatic tail was smaller (16.446±3.709 vs. 17.374±3.781), the positive enhanced integral was smaller (0.304±0.057 vs. 0.350±0.073) and the permeability surface was larger (27.823±8.228 vs. 21.509±7.768) (P<0.05). The effective radiation dose in the intermittent scan mode was slightly lower at 16.657±2.259 mSv than in the continuous scan mode (17.973±3.698 mSv). Conclusions: Different CT scanning intervals had a significant influence on whole pancreas blood volume, permeability surface, and positive enhanced integral. These demonstrate the high sensitivity of intermittent perfusion scanning for identifying perfusion abnormalities. Therefore, for the diagnosis of pancreatic diseases, intermittent pancreatic CT perfusion may be more advantageous.

YKL-40 derived from infiltrating macrophages cooperates with GDF15 to establish an immune suppressive microenvironment in gallbladder cancer.

Despite of the high lethality of gallbladder cancer (GBC), little is known regarding molecular regulation of the tumor immunosuppressive microenvironment. Here, we determined tumor expression levels of YKL-40 and the molecular mechanisms by which YKL-40 regulates escape of anti-tumor immune surveillance. We found that elevated expression levels of YKL-40 in plasma and tissue were correlated with tumor size, stage IV and lymph node metastasis. Single cell transcriptome analysis revealed that YKL-40 was predominantly derived from M2-like subtype of infiltrating macrophages. Blockade of M2-like macrophage differentiation of THP-1 cells with YKL-40 shRNA resulted in reprogramming to M1-like macrophages and restricting tumor development. YKL-40 induced tumor cell expression and secretion of growth differentiation factor 15 (GDF15), thus coordinating to promote PD-L1 expression mediated by PI3K, AKT and/or Erk activation. Interestingly, extracellular GDF15 inhibited intracellular expression of GDF15 that suppressed PD-L1 expression. Thus, YKL-40 disrupted the balance of pro- and anti-PD-L1 regulation to enhance expression of PD-L1 and inhibition of T cell cytotoxicity, leading to tumor immune evasion. The data suggest that YKL-40 and GDF15 could serve as diagnostic biomarkers and immunotherapeutic targets for GBC.

Exploration of the Corrosion Behavior of Electroless Plated Ni-P Amorphous Alloys via X-ray Photoelectron Spectroscopy.

A Ni-P amorphous alloy was deposited on a low carbon steel substrate via electroless plating. Further, the prepared samples were crystallized under the high temperature with a range from 200 °C to 500 °C in air for 1 h. The crystallization process was studied via XRD, AFM, and XPS, and anodic electrochemical behavior was investigated by potentiostatic methods in a 3.5 wt% NaCl solution. The experimental results indicate that the diffusion, dissolution, and enrichment of the component elements in the Ni-P alloy are essential during crystallization because the various corrosion behaviors corresponding to Ni and P are directly affected. More importantly, under the 400 °C treatment, H2PO2- was enriched in the alloy, which effectively hinders the anodic dissolution of nickel and forms a complete adsorption layer on the surface of the alloy. Our results demonstrate that P can effectively block the anodic dissolution of Ni during the corrosion process, and the crystallization process can effectively promote the surface enrichment of P to improve the corrosion resistance of the coating.

Long-term exposure to genistein inhibits the proliferation of gallbladder cancer by downregulating the MCM complex.

Soy isoflavones are natural tyrosine kinase inhibitors closely associated with decreased morbidity and mortality of various tumors. The activation of tyrosine kinases such as ERBB2 is the mechanism by which cholecystitis transforms into gallbladder cancer (GBC), therefore, it is important to investigate the relationship between long-term exposure to soy isoflavones and the occurrence and progression of GBC. This case-control study (n = 85 pairs) found that the high level of plasma soy isoflavone-genistein (GEN) was associated with a lower risk of gallbladder cancer (≥326.00 ng/mL compared to ≤19.30 ng/mL, crude odds ratio 0.15, 95% CI 0.04-0.59; P for trend = 0.016), and that the level of GEN exposure negatively correlated with Ki67 expression in GBC tissue (n = 85). Consistent with these results, the proliferation of GBC cells was inhibited in the long-term exposure models of GEN in vitro and in vivo. The long-term exposure to GEN reduced the tyrosine kinase activity of ERBB2 and impaired the function of the PTK6-AKT-GSK3ß axis, leading to downregulation of the MCM complex in GBC cells. In summary, long-term exposure to GEN associated with soy products intake might play a certain role in preventing GBC and even inhibiting the proliferation of GBC cells.

Quantitative Assessment of Pancreatic Fat by Quantitative CT in Type 2 Diabetes Mellitus.

Objective: To characterize the pancreatic fat deposition (PFD) in patients with type 2 diabetes mellitus (T2DM) by quantitative computed tomography (QCT) and investigate the relationship between PFD and clinical metabolic parameters and islet function. Materials and Methods: A total of 150 patients with T2DM and 93 age-matched healthy subjects underwent QCT to quantify PFD were included. PFD and various biochemical parameters were correlated by statistical methods and multiple stepwise linear regression modeling. Results: PFD measured by QCT in the T2DM group was statistically higher than that in the healthy control group, and the pancreatic CT value was statistically lower than that in the control group. The QCT measured PFD was negatively correlated with the pancreatic CT values (P < 0.001), and positively correlated with triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), visceral fat area (VAT) and insulin resistance index (HOMA-IR) (P < 0.05) in the T2DM patients. Multiple stepwise linear regression analysis identified PFD as the dependent variable factor for T2DM. Conclusion: This study suggests QCT as a reliable technique in measuring PFD in T2DM. High PFD is positively correlated with the degree of insulin resistance and may play an important role in islet cell dysfunction in T2DM.

Retraction Notice to: Long Noncoding RNA OIP5-AS1 Promotes the Progression of Liver Hepatocellular Carcinoma via Regulating the hsa-miR-26a-3p/EPHA2 Axis.

[This retracts the article DOI: 10.1016/j.omtn.2020.05.032.].

Effectiveness of polyene phosphatidylcholine and its combination with other drugs in patients with liver diseases based on real-world research.

OBJECTIVES: Polyene phosphatidylcholine (PPC) is a widely used hepatoprotective drug. We aim to explore the effectiveness of PPC in patients with liver diseases based on real-world research, and compare with other hepatoprotective drugs. METHODS: This was a 'three-phase' retrospective study, including a descriptive study, a self-control case study, and a specific-disease cohort study. A total of 14,800 hospitalized patients were enrolled in phase I from 1 January 2015 to 1 January 2020, of which 793 patients using PPC alone were included for phase II and III. The major measurement of effectiveness analysis was the ALT level and its changes. Wilcoxon signed-rank test, Chi-square test, and Mann-Whitney U test were used. RESULTS: In patients without liver tumor, ALT level decreased after using PPC (p < 0.01), and the decrease in ALT level using PPC was greater than using glutathione or magnesium isoglycyrrhizinate alone (p = 0.044; p = 0.038). In patients without liver tumor but having abnormal liver function, the decrease in ALT level using PPC + glutathione was greater than using glutathione alone (p = 0.047). CONCLUSION: PPC had a beneficial effect on liver function in patients without liver tumor, and PPC could enhance the liver protective function of glutathione and magnesium isoglycyrrhizinate.

Application of spectral CT combined with perfusion scan in diagnosis of pancreatic neuroendocrine tumors.

BACKGROUND: Pancreatic neuroendocrine tumors (pNETs) are heterogeneous tumors from the pancreatic neuroendocrine system, and early diagnosis is important for tumor prognosis and treatment. In this study, we aimed to explore the diagnostic value of spectral CT combined with perfusion scanning in improving the detection rate of pNETs. METHODS: From December 2018 to December 2020, 58 patients with clinically suspected pNETs were prospectively enrolled in the study for one-stop spectral CT combined with perfusion scanning, 36 patients were confirmed with pNETs by histopathology. An independent cohort of 30 patients with pNETs who underwent routine pancreatic perfusion scanning in our hospital during the same period were retrospectively collected. The image characters of pNETs versus tumor-free pancreatic parenchymal were examined. RESULTS: The detection rate of spectral CT combined with perfusion was 83.1-96.2%. CT values of the pNETs lesions under each single energy in the arterial phase were statistically higher than those of the adjacent normal pancreatic parenchyma. IC, WC and NIC, in the arterial phase of pNETs lesion were all statistically higher than those of the adjacent normal pancreatic parenchyma. The perfusion parameters of pNETs including BF, BV and MSI were significantly higher than those in normal parenchyma. The average effective radiation dose during the perfusion combined energy spectrum enhanced scanning process was 17.51 ± 2.18 mSv. CONCLUSION: The one-stop spectral CT combined with perfusion scan improves the detection of pNETs according to morphological features, perfusion parameters and energy spectrum characters with a relatively small radiation dose.

Novel protein kinase inhibitor TT-00420 inhibits gallbladder cancer by inhibiting JNK/JUN-mediated signaling pathway.

PURPOSE: This study aimed to investigate the efficiency of our chemically synthesized TT-00420, a novel spectrum-selective multiple protein kinase inhibitor, in cultured cells and animal models of gallbladder cancer (GBC) and explore its potential mechanism. METHODS: Multiple GBC models were established to assess the anti-tumor efficiency, toxicity, and pharmacokinetics of TT-00420. Integrated transcriptomic, proteomic and phosphoproteomic analysis was conducted to identify potential downstream effectors of TT-00420. Western blotting, qRT-PCR, nuclear-cytoplasm separation, and immunofluorescence were performed to confirm the multi-omic results and explore the molecular mechanism of TT-00420. Immunohistochemistry was used to detect FGFR1 and p-FGFR1 expression levels in GBC samples. Autodock software was utilized to investigate the potential binding mode between the TT-00420 and the human FGFR1. RESULTS: We found that TT-00420 exerted potent growth inhibition of GBC cell lines and multiple xenograft models. Treatment of mice with 15 mg/kg TT-00420 via gavage displayed a half-life of 1.8 h in the blood and rapid distribution to the liver, kidneys, lungs, spleen, and tumors at 0.25 h, but no toxicity to these organs over 2 weeks. Multi-omic analysis revealed c-Jun as a potential downstream effector after TT-00420 treatment. Mechanistically, TT-00420 showed rigorous ability to block FGFR1 and its downstream JNK-JUN (S63/S73) signaling pathway, and induce c-Jun S243-dependent MEK/ERK reactivation, leading to FASLG-dependent tumor cell death. Finally, we found that FGFR1 and p-FGFR1 expression was elevated in GBC patients and these levels correlated with decreased patient survival. CONCLUSIONS: TT-00420 shows potent antitumor efficacy and may serve as a novel agent to improve GBC prognosis.

Understanding the P-Loop Conformation in the Determination of Inhibitor Selectivity Toward the Hepatocellular Carcinoma-Associated Dark Kinase STK17B.

As a member of the death-associated protein kinase family of serine/threonine kinases, the STK17B has been associated with diverse diseases such as hepatocellular carcinoma. However, the conformational dynamics of the phosphate-binding loop (P-loop) in the determination of inhibitor selectivity profile to the STK17B are less understood. Here, a multi-microsecond length molecular dynamics (MD) simulation of STK17B in the three different states (ligand-free, ADP-bound, and ligand-bound states) was carried out to uncover the conformational plasticity of the P-loop. Together with the analyses of principal component analysis, cross-correlation and generalized correlation motions, secondary structural analysis, and community network analysis, the conformational dynamics of the P-loop in the different states were revealed, in which the P-loop flipped into the ADP-binding site upon the inhibitor binding and interacted with the inhibitor and the C-lobe, strengthened the communication between the N- and C-lobes. These resulting interactions contributed to inhibitor selectivity profile to the STK17B. Our results may advance our understanding of kinase inhibitor selectivity and offer possible implications for the design of highly selective inhibitors for other protein kinases.

A Vimentin-Targeting Oral Compound with Host-Directed Antiviral and Anti-Inflammatory Actions Addresses Multiple Features of COVID-19 and Related Diseases.

Damage in COVID-19 results from both the SARS-CoV-2 virus and its triggered overactive host immune responses. Therapeutic agents that focus solely on reducing viral load or hyperinflammation fail to provide satisfying outcomes in all cases. Although viral and cellular factors have been extensively profiled to identify potential anti-COVID-19 targets, new drugs with significant efficacy remain to be developed. Here, we report the potent preclinical efficacy of ALD-R491, a vimentin-targeting small molecule compound, in treating COVID-19 through its host-directed antiviral and anti-inflammatory actions. We found that by altering the physical properties of vimentin filaments, ALD-491 affected general cellular processes as well as specific cellular functions relevant to SARS-CoV-2 infection. Specifically, ALD-R491 reduced endocytosis, endosomal trafficking, and exosomal release, thus impeding the entry and egress of the virus; increased the microcidal capacity of macrophages, thus facilitating the pathogen clearance; and enhanced the activity of regulatory T cells, therefore suppressing the overactive immune responses. In cultured cells, ALD-R491 potently inhibited the SARS-CoV-2 spike protein and human ACE2-mediated pseudoviral infection. In aged mice with ongoing, productive SARS-CoV-2 infection, ALD-R491 reduced disease symptoms as well as lung damage. In rats, ALD-R491 also reduced bleomycin-induced lung injury and fibrosis. Our results indicate a unique mechanism and significant therapeutic potential for ALD-R491 against COVID-19. We anticipate that ALD-R491, an oral, fast-acting, and non-cytotoxic agent targeting the cellular protein with multipart actions, will be convenient, safe, and broadly effective, regardless of viral mutations, for patients with early- or late-stage disease, post-COVID-19 complications, and other related diseases. IMPORTANCE With the Delta variant currently fueling a resurgence of new infections in the fully vaccinated population, developing an effective therapeutic drug is especially critical and urgent in fighting COVID-19. In contrast to the many efforts to repurpose existing drugs or address only one aspect of COVID-19, we are developing a novel agent with first-in-class mechanisms of action that address both the viral infection and the overactive immune system in the pathogenesis of the disease. Unlike virus-directed therapeutics that may lose efficacy due to viral mutations, and immunosuppressants that require ideal timing to be effective, this agent, with its unique host-directed antiviral and anti-inflammatory actions, can work against all variants of the virus, be effective during all stages of the disease, and even resolve post-disease damage and complications. Further development of the compound will provide an important tool in the fight against COVID-19 and its complications, as well as future outbreaks of new viruses.

Exosomal microRNA-15a from mesenchymal stem cells impedes hepatocellular carcinoma progression via downregulation of SALL4.

Hepatocellular carcinoma (HCC) is a heterogeneous tumor with an increased incidence worldwide accompanied by high mortality and dismal prognosis. Emerging evidence indicates that mesenchymal stem cells (MSCs)-derived exosomes possess protective effects against various human diseases by transporting microRNAs (miRNAs or miRs). We aimed to explore the role of exosomal miR-15a derived from MSCs and its related mechanisms in HCC. Exosomes were isolated from transduced MSCs and co-incubated with Hep3B and Huh7 cells. miR-15a expression was examined by RT-qPCR in HCC cells, MSCs, and secreted exosomes. CCK-8, transwell, and flow cytometry were used to detect the effects of miR-15a or spalt-like transcription factor 4 (SALL4) on cell proliferative, migrating, invasive, and apoptotic properties. A dual-luciferase reporter gene assay was performed to validate the predicted targeting relationship of miR-15a with SALL4. Finally, in vivo experiments in nude mice were implemented to assess the impact of exosome-delivered miR-15a on HCC. The exosomes from MSCs restrained HCC cell proliferative, migrating, and invasive potentials, and accelerated their apoptosis. miR-15a was expressed at low levels in HCC cells and could bind to SALL4, thus curtailing the proliferative, migrating, and invasive abilities of HCC cells. Exosomes successfully delivered miR-15a to HCC cells. Exosomal miR-15a depressed tumorigenicity and metastasis of HCC tumors in vivo. Overall, exosomal miR-15a from MSCs can downregulate SALL4 expression and thereby retard HCC development.

Albiflorin alleviates cognitive dysfunction in STZ-induced rats.

BACKGROUND: To explore the effect of albiflorin (AL) on streptozotocin (STZ)-induced Alzheimer's disease (AD) in rats. METHODS: A mouse model of diabetic encephalopathy was established by intraperitoneal injection of 1%STZ. Step down test and water maze test were used to test the cognitive function of rats. Congo Red Staining was used to detect the distribution of Aß plaques in the hippocampus of rats. Cytokine levels in serum and hippocampus were measured using ELISA. Serum insulin, oral glucose tolerance (OGTT), serum superoxide dismutase (SOD) activity and malondialdehyde (MDA) content were measured by commercial kits. And the content of Nrf-2/HO-1/HMGB1/NF-kB in the hippocampus of diabetic rats were detected by western blot. RESULTS AND CONCLUSION: Compared with the STZ model group, the average escape latency of rats in the AL group in the Morris water maze test was significantly shortened, and the average number of platform crossings and the ratio of distance/total swimming distance in the target quadrant were increased significantly. Staining of tissue sections and ELISA showed a decrease in Aß plaque density in the hippocampus of rats in the AL group. And serum insulin levels of rats in the ALgroup were significantly reduced and OGTT was improved. In addition, AL could also regulate the Nrf-2/HO-1/HMGB1/NF-kB signal pathway in the hippocampus. Therefore, AL may ameliorate STZ-induced cognitive impairment in rats by regulating oxidative stress and inflammation in the brain.