Design, synthesis and biological evaluation of 4-(indolin-1-yl)-6-substituted-pyrido[3,2-d]pyrimidine derivatives as Mnk1/2 inhibitors.

The Mnk-eIF4E axis plays a crucial role in tumor development, and inhibiting Mnk kinases is a promising approach for cancer therapy. Starting with fragment WS23, a series of 4-(indolin-1-yl)-6-substituted-pyrido[3,2-d]pyrimidine derivatives were designed and synthesized. Among these derivatives, compound 15b showed the highest potency with IC50 values of 0.8 and 1.5 nM against Mnk1 and Mnk2, respectively. Additionally, it demonstrated good selectivity among 30 selected kinases. 15b significantly suppressed MOLM-13 and K562 cell lines growth and caused cell cycle arrest. Furthermore, the Western blot assay revealed that 15b effectively downregulated the downstream proteins p-eIF4E, Mcl-1, and c-myc. Additionally, 15b exhibited remarkable stability in rat plasma and rat and human microsomes. In vivo anti-tumor activity study suggested that treatment with 15b suppressed tumor growth in LL/2 syngeneic models. These findings highlight the potential of 15b as a novel and potent Mnks inhibitor, which deserves further investigation.

Real-world study of hepatic artery infusion chemotherapy combined with anti-PD-1 immunotherapy for hepatocellular carcinoma patients with portal vein tumor thrombus.

Background: Patients with hepatocellular carcinoma (HCC) with portal vein tumor thrombus (PVTT) present a poor prognosis. Current systemic therapies offer limited benefits. Hepatic artery infusion chemotherapy (HAIC) is a local regional treatment for advanced HCC, particularly in selected patients such as patients with PVTT or high intrahepatic tumor burden. Objectives: The purpose of this study is to retrospectively evaluate the efficacy and safety of HAIC combined with anti-PD-1 immunotherapy for HCC patients with PVTT, and explore factors related to survival prognosis, providing clues for treatment decisions for HCC patients. Design: This is a single-center retrospective study conducted over 2 years on consecutive PVTT patients receiving HAIC combined anti-PD-1 antibodies. Methods: The primary endpoint was overall survival (OS). Univariate and multivariate analyses were performed to identify prognostic factors affecting OS. Treatment-associated adverse events were evaluated as well. Results: A total of 119 patients were analyzed. The median OS and PFS were 14.9 months and 6.9 months. A total of 31.1% of grade 3-4 adverse events were reported, with elevated transaminase and total bilirubin being the most common. The independent variables correlated with survival include treatment-related alpha-fetoprotein (AFP) response, the presence of extrahepatic organ metastasis, absolute value of platelet (PLT), neutrophil-to-lymphocyte ratio, and combined usage of tyrosine kinase inhibitors (TKIs). Conclusion: In HCC patients with PVTT, combination therapy with HAIC and anti-PD-1 antibodies might be a promising therapy. The efficacy and safety of this combination protocol on patients with HCC complicated by PVTT warrants further investigation prospectively, especially in combination with TKIs.

Development and Preliminary Validation of a Novel Convolutional Neural Network Model for Predicting Treatment Response in Patients with Unresectable Hepatocellular Carcinoma Receiving Hepatic Arterial Infusion Chemotherapy.

The goal of this study was to evaluate the performance of a convolutional neural network (CNN) with preoperative MRI and clinical factors in predicting the treatment response of unresectable hepatocellular carcinoma (HCC) patients receiving hepatic arterial infusion chemotherapy (HAIC). A total of 191 patients with unresectable HCC who underwent HAIC in our hospital between May 2019 and March 2022 were retrospectively recruited. We selected InceptionV4 from three representative CNN models, AlexNet, ResNet, and InceptionV4, according to the cross-entropy loss (CEL). We subsequently developed InceptionV4 to fuse the information from qualified pretreatment MRI data and patient clinical factors. Radiomic information was evaluated based on several constant sequences, including enhanced T1-weighted sequences (with arterial, portal, and delayed phases), T2 FSE sequences, and dual-echo sequences. The performance of InceptionV4 was cross-validated in the training cohort (n = 127) and internally validated in an independent cohort (n = 64), with comparisons against single important clinical factors and radiologists in terms of receiver operating characteristic (ROC) curves. Class activation mapping was used to visualize the InceptionV4 model. The InceptionV4 model achieved an AUC of 0.871 (95% confidence interval [CI] 0.761-0.981) in the cross-validation cohort and an AUC of 0.826 (95% CI 0.682-0.970) in the internal validation cohort; these two models performed better than did the other methods (AUC ranges 0.783-0.873 and 0.708-0.806 for cross- and internal validations, respectively; P < 0.01). The present InceptionV4 model, which integrates radiomic information and clinical factors, helps predict the treatment response of unresectable HCC patients receiving HAIC treatment.

LINC02362/hsa-miR-18a-5p/FDX1 axis suppresses proliferation and drives cuproptosis and oxaliplatin sensitivity of hepatocellular carcinoma.

Cuproptosis is a novel cell death mechanism caused by copper overload, with FDX1 serving as the key regulator. LncRNAs are known to play a significant role in the aberrant regulation of gene expression in hepatocellular carcinoma (HCC). In this study, we investigated the biological role of the LINC02362/hsa-miR-18a-5p/FDX1 axis in HCC. We first explored the expression pattern, prognostic value, biological functions, drug sensitivity, and immune effect of FDX1. Using bioinformatics techniques, we then predicted several potential target lncRNAs and miRNAs. We identified a lncRNA-miRNA-FDX1 axis based on the ceRNA mechanism. In vitro experiments were conducted to validate the relationship between the lncRNA-miRNA-FDX1 axis and its biological effects in HCC. Finally, we investigated the relationship between the LINC02362/hsa-miR-18a-5p/FDX1 axis and oxaliplatin-induced cuproptosis in HCC. Our findings indicated that FDX1 expression was downregulated in HCC tissues; however, elevated FDX1 expression correlates with improved prognosis and heightened sensitivity to oxaliplatin. We confirmed that LINC02362 binds to and directly regulates the expression of miR-18a-5p, with FDX1 a target of miR-18a-5p. Experimental results suggested that upregulating LINC02362/hsa-miR-18a-5p/FDX1 axis suppressed the proliferation of HCC cells. Furthermore, LINC02362 knockdown led to a reduction in copper concentration and resistance to elesclomol-Cu. We also discovered that augmenting the LINC02362/hsa-miR-18a-5p/FDX1 axis could bolster the sensitivity of HCC to oxaliplatin through cuproptosis. This work presents the LINC02362/hsa-miR-18a-5p/FDX1 axis as a novel pathway that triggers cuproptosis and enhances the sensitivity of HCC to oxaliplatin, presenting a promising therapeutic avenue to combat oxaliplatin resistance in HCC.

Gene Cloning and Characterization of Transcription Factor FtNAC10 in Tartary Buckwheat (Fagopyrum tataricum (L.) Gaertn.).

NAC transcription factors play a significant role in plant stress responses. In this study, an NAC transcription factor, with a CDS of 792 bp encoding 263 amino acids, was cloned from Fagopyrum tataricum (L.) Gaertn. (F. tataricum), a minor cereal crop, which is rich in flavonoids and highly stress resistant. The transcription factor was named FtNAC10 (NCBI accession number: MK614506.1) and characterized as a member of the NAP subgroup of NAC transcriptions factors. The gene exhibited a highly conserved N-terminal, encoding about 150 amino acids, and a highly specific C-terminal. The resulting protein was revealed to be hydrophilic, with strong transcriptional activation activity. FtNAC10 expression occurred in various F. tataricum tissues, most noticeably in the root, and was regulated differently under various stress treatments. The over-expression of FtNAC10 in transgenic Arabidopsis thaliana (A. thaliana) seeds inhibited germination, and the presence of FtNAC10 enhanced root elongation under saline and drought stress. According to phylogenetic analysis and previous reports, our experiments indicate that FtNAC10 may regulate the stress response or development of F. tataricum through ABA-signaling pathway, although the mechanism is not yet known. This study provides a reference for further analysis of the regulatory function of FtNAC10 and the mechanism that underlies stress responses in Tartary buckwheat.

Characterization of the complete chloroplast genome of Elymus alashanicus (Keng) S. L. Chen, and its phylogenetic analysis.

Elymus alashanicus (Keng) S. L. Chen, a herbaceous plant endemic to China, plays a crucial role in the local ecosystems. In this study, we sequenced and characterized the complete chloroplast (cp) genome of E. alashanicus, which is 135,072 bp in length and arranged in a circular form. The cp genome includes a pair of inverted repeats (IRa and IRb) of 20,813 bp each, separated by a large single-copy (LSC) region of 80,678 bp and a small single-copy (SSC) region of 12,768 bp. The cp genome contains 130 genes, including 83 protein-coding genes, 39 tRNA genes, and eight rRNA genes. Phylogenetic analysis revealed that E. alashanicus is closely related to Elymus breviaristatus and Campeiostachys dahurica var. tangutorum in current sampling. Our findings provide valuable insights into the cp genome of E. alashanicus, which could contribute to further studies on the evolution and conservation of this species.

Hypoxia-induced reprogramming of glucose-dependent metabolic pathways maintains the stemness of human bone marrow-derived endothelial progenitor cells.

The benefits of hypoxia for maintaining the stemness of cultured human bone marrow-derived endothelial progenitor cells (BM EPCs) have previously been demonstrated but the mechanisms responsible remain unclear. Growing evidences suggest that cellular metabolism plays an important role in regulating stem cell fate and self-renewal. Here we aimed to detect the changes of glucose metabolism and to explore its role on maintaining the stemness of BM EPCs under hypoxia. We identified the metabolic status of BM EPCs by using extracellular flux analysis, LC-MS/MS, and 13C tracing HPLC-QE-MS, and found that hypoxia induced glucose metabolic reprogramming, which manifested as increased glycolysis and pentose phosphate pathway (PPP), decreased tricarboxylic acid (TCA) and mitochondrial respiration. We further pharmacologically altered the metabolic status of cells by employing various of inhibitors of key enzymes of glycolysis, PPP, TCA cycle and mitochondria electron transport chain (ETC). We found that inhibiting glycolysis or PPP impaired cell proliferation either under normoxia or hypoxia. On the contrary, inhibiting pyruvate oxidation, TCA or ETC promoted cell proliferation under normoxia mimicking hypoxic conditions. Moreover, promoting pyruvate oxidation reverses the maintenance effect of hypoxia on cell stemness. Taken together, our data suggest that hypoxia induced glucose metabolic reprogramming maintains the stemness of BM EPCs, and artificial manipulation of cell metabolism can be an effective way for regulating the stemness of BM EPCs, thereby improving the efficiency of cell expansion in vitro.

Phosphatase regenerating liver 3 participates in Integrinß1/FAK-Src/MAPK signaling pathway and contributes to the regulation of malignant behaviors in hepatocellular carcinoma cells.

Background: Hepatocellular carcinoma (HCC) is the leading cause of mortality worldwide. Phosphatase regenerating liver 3 (PRL-3) was associated with cancer metastasis. However, the significance of PRL-3 in the prognosis of HCC remains elusive. The aim of this study was to elucidate the role of PRL-3 in HCC metastasis and its prognosis. Methods: The expressions of PRL-3 in cancer tissues isolated from 114 HCC patients, who underwent curative hepatectomy from May to November in 2008, were analyzed by immunohistochemistry, and its prognostic significance was evaluated. Thereafter, the migration, invasion, and metastatic alterations in MHCC97H cells with PRL-3 overexpression or knockdown were explored and compared with the tumor size and lung metastasis in orthotopic HCC model of nude mice derived from MHCC97H cells with PRL-3 overexpression or knockdown. The underlying mechanism involving PRL-3-mediated effect on HCC migration, invasion, and metastasis was further examined. Results: Univariate and multivariate analysis demonstrated PRL-3 overexpression was an independent prognostic factor for poor overall survival (OS) and progression-free survival (PFS) of the HCC patients. Increased PRL-3 expression in MHCC97H cells was in accordance with the enhanced metastasis potential. PRL-3 knockdown inhibited the migration, invasiveness, and clone forming ability in MHCC97H cells, whereas PRL-3 overexpression reverted the above behavior. The growth of xenograft tumor in the liver was suppressed, and the lung metastasis in nude mice was inhibited by PRL-3 downregulation. The knockdown of PRL-3 could downregulate the expressions of Integrinß1 and p-Src (Tyr416), p-Erk (Thr202/Tyr204) activation, and reduce MMP9 expression. Both MEK1/2 inhibitor (U0126) and Src inhibitor could repress PRL-3-induced invasiveness and migration in MHCC97H cells. Conclusions: PRL-3 was significantly overexpressed and an independent prognostic factor to predict the death of HCC patients. Mechanically, PRL-3 plays a critical role in HCC invasive and metastasis via Integrinß1/FAK-Src/RasMAPK signaling. Validation of PRL-3 as a clinical prediction marker in HCC warrants further research.

Hydroxysafflor yellow a confers neuroprotection against acute traumatic brain injury by modulating neuronal autophagy to inhibit NLRP3 inflammasomes.

ETHNOPHARMACOLOGICAL RELEVANCE: Hydroxysafflor yellow A (HSYA) is the principal bioactive compound isolated from the plant Carthamus tinctorius L. and has been reported to exert neuroprotective effects against various neurological diseases, including traumatic brain injury (TBI). However, the specific molecular and cellular mechanisms underlying HSYA-mediated neuroprotection against TBI are unclear. AIM OF THE STUDY: This study explored the effects of HSYA on autophagy and the NLRP3 inflammasome in mice with TBI and the related mechanisms. MATERIALS AND METHODS: Mice were subjected to TBI and treated with or without HSYA. Neurological severity scoring, LDH assays and apoptosis detection were first performed to assess the effects of HSYA in mice with TBI. RNA-seq was then conducted to explore the mechanisms that contributed to HSYA-mediated neuroprotection. ELISA, western blotting, and immunofluorescence were performed to further investigate the mechanisms of neuroinflammation and autophagy. Moreover, 3-methyladenine (3-MA), an autophagy inhibitor, was applied to determine the connection between autophagy and the NLRP3 inflammasome. RESULTS: HSYA significantly decreased the neurological severity score, serum LDH levels and apoptosis in mice with TBI. A total of 921 differentially expressed genes were identified in the cortices of HSYA-treated mice with TBI and were significantly enriched in the inflammatory response and autophagy. Furthermore, HSYA treatment markedly reduced inflammatory cytokine levels and astrocyte activation. Importantly, HSYA suppressed neuronal NLRP3 inflammasome activation, as indicated by decreased levels of NLRP3, ASC and cleaved caspase-1 and a reduced NLRP3+ neuron number. It increased autophagy and ameliorated autophagic flux dysfunction, as evidenced by increased LC3 II/LC3 I levels and decreased P62 levels. The effects of HSYA on the NLRP3 inflammasome were abolished by 3-MA. Mechanistically, HSYA may enhance autophagy through AMPK/mTOR signalling. CONCLUSION: HSYA enhanced neuronal autophagy by triggering the AMPK/mTOR signalling pathway, leading to inhibition of the NLRP3 inflammasome to improve neurological recovery after TBI.

Characteristics and candidate genes associated with excellent stalk strength in maize (Zea mays L.).

Lodging is a major problem in maize production, which seriously affects yield and hinders mechanized harvesting. Improving stalk strength is an effective way to improve lodging. The maize inbred line Jing2416 (J2416) was an elite germplasm in maize breeding which had strong stalk mechanical strength. To explore the characteristics its stalk strength, we conducted physiological, metabolic and transcriptomic analyses of J2416 and its parents Jing24 (J24) and 5237. At the kernel dent stage, the stalk rind penetrometer strength of J2416 was significantly higher than those of its two parents in multiple environments. The rind thickness, sclerenchyma tissue thickness, and cellulose, hemicellulose, and lignin contents of J2416 were significantly higher than those of its parents. Based on the significant differences between J2416 and 5237, we detected metabolites and gene transcripts showing differences in abundance between these two materials. A total of 212 (68.60%) metabolites and 2287 (43.34%) genes were up-regulated in J2416 compared with 5237. The phenylpropanoid and glycan synthesis/metabolism pathways were enriched in metabolites and genes that were up-regulated in J2416. Twenty-eight of the up-regulated genes in J2416 were involved in lignin, cellulose, and hemicellulose synthesis pathways. These analyses have revealed important physiological characteristics and candidate genes that will be useful for research and breeding of inbred lines with excellent stalk strength.

Predicting Disease-Specific Survival for Patients With Primary Cholangiocarcinoma Undergoing Curative Resection by Using a Decision Tree Model.

Background: The aim of this study was to derive and validate a decision tree model to predict disease-specific survival after curative resection for primary cholangiocarcinoma (CCA). Method: Twenty-one clinical characteristics were collected from 482 patients after curative resection for primary CCA. A total of 289 patients were randomly allocated into a training cohort and 193 were randomly allocated into a validation cohort. We built three decision tree models based on 5, 12, and 21 variables, respectively. Area under curve (AUC), sensitivity, and specificity were used for comparison of the 0.5-, 1-, and 3-year decision tree models and regression models. AUC and decision curve analysis (DCA) were used to determine the predictive performances of the 0.5-, 1-, and 3-year decision tree models and AJCC TNM stage models. Results: According to the fitting degree and the computational cost, the decision tree model derived from 12 variables displayed superior predictive efficacy to the other two models, with an accuracy of 0.938 in the training cohort and 0.751 in the validation cohort. Maximum tumor size, resection margin, lymph node status, histological differentiation, TB level, ALBI, AKP, AAPR, ALT, γ-GT, CA19-9, and Child-Pugh grade were involved in the model. The performances of 0.5-, 1-, and 3-year decision tree models were better than those of conventional models and AJCC TNM stage models. Conclusion: We developed a decision tree model to predict outcomes for CCA undergoing curative resection. The present decision tree model outperformed other clinical models, facilitating individual decision-making of adjuvant therapy after curative resection.

Freehand Minimally Invasive Pedicle Screw Fixation and Minimally Invasive Decompression for a Thoracic or Lumbar Vertebral Metastatic Tumor From Hepatocellular Carcinoma.

Objective: To compare freehand minimally invasive pedicle screw fixation (freehand MIPS) combined with percutaneous vertebroplasty (PVP), minimally invasive decompression, and partial tumor resection with open surgery for treatment of thoracic or lumbar vertebral metastasis of hepatocellular carcinoma (HCC) with symptoms of neurologic compression, and evaluate its feasibility, efficacy, and safety. Methods: Forty-seven patients with 1-level HCC metastatic thoracolumbar tumor and neurologic symptoms were included between February 2015 and April 2017. Among them, 21 patients underwent freehand MIPS combined with PVP, minimally invasive decompression, and partial tumor resection (group 1), while 26 patients were treated with open surgery (group 2). Duration of operation, blood loss, times of fluoroscopy, incision length, and stay in hospital were compared between the two groups. Pre- and postoperative visual analog scale (VAS) pain score, Oswestry Disability Index (ODI), American Spinal Injury Association (ASIA) grade, ambulatory status, and urinary continence were also recorded. The Cobb angle and central and anterior vertebral body height were measured on lateral radiographs before surgery and during follow-ups. Results: Patients in group 1 showed significantly less blood loss (195.5 ± 169.1 ml vs. 873.1 ± 317.9 ml, P = 0.000), shorter incision length (3.4 ± 0.3 vs. 13.6 ± 1.8 cm, P = 0.000), shorter median stay in hospital (4-8/6 vs. 8-17/12 days, P = 0.000), more median times of fluoroscopy (5-11/6 vs. 4-7/5 times, P = 0.000), and longer duration of operation (204.8 ± 12.1 vs. 171.0 ± 12.0 min, P = 0.000) than group 2. Though VAS significantly decreased after surgery in both groups, VAS of group 1 was significantly lower than that of group 2 immediately after surgery and during follow-ups (P < 0.05). Similar results were found in ODI. No differences in the neurological improvement and spinal stability were observed between the two groups. Conclusion: Freehand MIPS combined with PVP, minimally invasive decompression, and partial tumor resection is a safe, effective, and minimally invasive method for treating thoracolumbar metastatic tumors of HCC, with less blood loss, better pain relief, and shorter length of midline incision and stay in hospital.

The Prognostic Value of the Albumin to Gamma-Glutamyltransferase Ratio in Patients with Hepatocellular Carcinoma Undergoing Radiofrequency Ablation.

Albumin to gamma-glutamyltransferase ratio (AGR) is a newly developed biomarker for the prediction of patients' prognosis in solid tumors. The purpose of the study was to establish a novel AGR-based nomogram to predict tumor prognosis in patients with early-stage HCC undergoing radiofrequency ablation (RFA). 394 hepatocellular carcinoma (HCC) patients who had received RFA as initial treatment were classified into the training cohort and validation cohort. Independent prognostic factors were identified by univariate and multivariate analyses. The value of AGR was evaluated by the concordance index (C-index), receiver operating characteristic (ROC) curves, and likelihood ratio tests (LAT). Logistic regression and nomogram were performed to establish the pretreatment scoring model based on the clinical variables. As a result, AGR = 0.63 was identified as the best cutoff value to predict overall survival (OS) in the training cohort. According to the results of multivariate analysis, AGR was an independent indicator for OS and recurrence-free survival (RFS). In both training cohort and validation cohort, the high-AGR group showed better RFS and OS than the low-AGR group. What is more, the C-index, area under the ROC curves, and LAT χ 2 values suggested that AGR outperformed the Child-Pugh (CP) grade and albumin-bilirubin (ALBI) grade in terms of predicting OS. The AGR, AKP, and tumor size were used to establish the OS nomogram. Besides, the results of Hosmer-Lemeshow test and calibration curve analysis displayed that both nomograms in the training and validation cohorts performed well in terms of calibration. Therefore, the AGR-based nomogram can predict the postoperative prognosis of early HCC patients undergoing RFA.

Development of a Prognostic Scoring System for Hepatocellular Carcinoma Patients With Main Portal Vein Tumor Thrombus Undergoing Conventional Transarterial Chemoembolization: An Analysis of 173 Patients.

BACKGROUND: Patients with hepatocellular carcinoma (HCC) with main portal vein tumor thrombus (mPVTT) have poor prognosis. Promising systemic therapies, such as target therapies, have limited benefits. The purpose of this study is to retrospectively evaluate the benefits of conventional TACE (c-TACE) and to establish a prognostic stratification of HCC patients with mPVTT. METHODS: This is a single center retrospective study conducted over 5 years (duration of performing c-TACE), on consecutive HCC patients with mPVTT receiving c-TACE. Univariable and multivariable analysis were used to explore factors independently associated with overall survival (OS). Based on Cox-regression analysis, prognostic models were developed and internally validated by bootstrap methods. Discrimination and performance were measured by Akaike information criterion, concordance index, and likelihood ratio test. RESULTS: A total of 173 patients were included. Median OS was 6.0 months (95%CI: 3.92~8.08). The independent variables correlated with survival were largest tumor diameter, tumor number, mPVTT extension, and AFP. In the final model, patients were assigned 2 points if largest tumor diameter ≥8 cm, or tumor number ≥2, 1point if main trunk was complete obstructed, or AFP ≥400 ng/ml. By summing up these points, patients were divided into three risk groups according to the score at the 15rd and 85th percentiles, in which median OS were 18, 7, and 3.5months, respectively (p<0.001). The model shown optimal discrimination, performance, and calibration. CONCLUSIONS: c-TACE could provide survival benefits in HCC patients with mPVTT and the proposed prognostic stratification may help to identify good candidates for the treatment, and those for whom c-TACE may be futile.

Self-consistent kinetic model of nested electron- and ion-scale magnetic cavities in space plasmas.

NASA's Magnetospheric Multi-Scale (MMS) mission is designed to explore the proton- and electron-gyroscale kinetics of plasma turbulence where the bulk of particle acceleration and heating takes place. Understanding the nature of cross-scale structures ubiquitous as magnetic cavities is important to assess the energy partition, cascade and conversion in the plasma universe. Here, we present theoretical insight into magnetic cavities by deriving a self-consistent, kinetic theory of these coherent structures. By taking advantage of the multipoint measurements from the MMS constellation, we demonstrate that our kinetic model can utilize magnetic cavity observations by one MMS spacecraft to predict measurements from a second/third spacecraft. The methodology of "observe and predict" validates the theory we have derived, and confirms that nested magnetic cavities are self-organized plasma structures supported by trapped proton and electron populations in analogous to the classical theta-pinches in laboratory plasmas.

Annotation of genes involved in high level of dihydromyricetin production in vine tea (Ampelopsis grossedentata) by transcriptome analysis.

BACKGROUND: Leaves of the medicinal plant Ampelopsis grossedentata, which is commonly known as vine tea, are used widely in the traditional Chinese beverage in southwest China. The leaves contain a large amount of dihydromyricetin, a compound with various biological activities. However, the transcript profiles involved in its biosynthetic pathway in this plant are unknown. RESULTS: We conducted a transcriptome analysis of both young and old leaves of the vine tea plant using Illumina sequencing. Of the transcriptome datasets, a total of 52.47 million and 47.25 million clean reads were obtained from young and old leaves, respectively. Among 471,658 transcripts and 177,422 genes generated, 7768 differentially expressed genes were identified in leaves at these two stages of development. The phenylpropanoid biosynthetic pathway of vine tea was investigated according to the transcriptome profiling analysis. Most of the genes encoding phenylpropanoid biosynthesis enzymes were identified and found to be differentially expressed in different tissues and leaf stages of vine tea and also greatly contributed to the biosynthesis of dihydromyricetin in vine tea. CONCLUSIONS: To the best of our knowledge, this is the first formal study to explore the transcriptome of A. grossedentata. The study provides an insight into the expression patterns and differential distribution of genes related to dihydromyricetin biosynthesis in vine tea. The information may pave the way to metabolically engineering plants with higher flavonoid content.

Evaluation of antioxidant properties of the different tissues of vine tea (Ampelopsis grossedentata) in stripped canola oil and sunflower oil.

The leaves of Ampelopsis grossedentata have a long history of use as a health tea and herbal medicine. Data on the distribution of active metabolites in, and antioxidant capacities of, different vine tea tissues remain incomplete. The aim of this work was to investigate the content of metabolites from A. grossedentata different tissues and evaluate the antioxidant capacities of the extraction of the species in vitro and in oil systems: canola oil and sunflower oil. To evaluate the degree of lipid oxidation, the peroxide value (POV) and thiobarbituric acid-reactive substance value (TBARS) were determined, and proton nuclear magnetic resonance (1 H-NMR) was performed. The results revealed a high total flavonoid content in each of the four extractions (>580 mg/g dried weigh). Leaf extractions exhibited higher antioxidant ability, followed by fruit extract and butylated hydroxytoluene (BHT). The POVs of oils bearing extracts of A. grossedentata and BHT maintained less than 21.08 meq/kg oil against to control with 1,406.33 ± 52.63 meq/kg oil on day 32 in canola oil, and 27.87 meq/kg oil comparing to 1,892.96 ± 48.63 meq/kg oil in control on day 24 in sunflower oil. Concurring results were also obtained in TBARS and 1 H-NMR analysis. Our results indicated that these different tissues of A. grossedentata could be a potential antioxidant resource, and this work may contribute to the comprehensive utilization of this species. PRACTICAL APPLICATION: Leaf extracts of Ampelopsis grossedentata showed effective antioxidant properties, followed by the fruit extract, which showed similar activity to that of the synthetic antioxidant of BHT. Moreover, the investigation of different tissues within the plant may contribute to the comprehensive utilization of this species.

Injectable double-crosslinked hydrogels with kartogenin-conjugated polyurethane nano-particles and transforming growth factor ß3 for in-situ cartilage regeneration.

Articular cartilage has a limited ability for self-repair after injury. Implantation of scaffolds functionalized with bioactive molecules that could induce the migration and chondrogenesis of endogenous mesenchymal stem cells (MSCs) provides a convenient alternative for in-situ cartilage regeneration. In this study, we found the synergistic effects of kartogenin (KGN) and transforming growth factor ß3 (TGF-ß3) on chondrogenesis of MSCs in vitro, indicating that KGN and TGF-ß3 are a good match for cartilage regeneration. Furthermore, we confirmed that KGN promoted the chondrogenesis of MSCs through attenuating the degradation of Runx1, which physically interacted with p-Smad3 in nuclei of MSCs. Meanwhile, we designed an injectable double-crosslinked hydrogel with superior mechanical property and longer support for cartilage regeneration by modifying sodium alginate and gelatin. When loaded with KGN conjugated polyurethane nanoparticles (PN-KGN) and TGF-ß3, this hydrogel showed biological functions by the release of KGN and TGF-ß3, which promoted the MSC migration and cartilage regeneration in one system. In conclusion, the cell-free hydrogel, along with PN-KGN and TGF-ß3, provides a promising strategy for cartilage repair by attracting endogenous MSCs and inducing chondrogenesis of recruited cells in a single-step procedure.

Alterations in exosomal miRNA profile upon epithelial-mesenchymal transition in human lung cancer cell lines.

BACKGROUND: Epithelial-mesenchymal transition (EMT) is regarded as a critical event during tumor metastasis. Recent studies have revealed changes and the contributions of proteins in/on exosomes during EMT. Besides proteins, microRNA (miRNA) is another important functional component of exosomes. We hypothesized that the miRNA profile of exosomes may change following EMT and these exosomal miRNAs may in return promote EMT, migration and invasion of cancer cells. RESULTS: The small RNA profile of exosomes was altered following EMT. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed that the specific miRNAs of M-exosomes have the potential to drive signal transduction networks in EMT and cancer progression. Co-culture experiments confirmed that M-exosomes can enter epithelial cells and promote migration, invasion and expression of mesenchymal markers in the recipient cells. CONCLUSION: Our results reveal changes in the function and miRNA profile of exosomes upon EMT. M-exosomes can promote transfer of the malignant (mesenchymal) phenotype to epithelial recipient cells. Further, the miRNAs specifically expressed in M-exosomes are associated with EMT and metastasis, and may serve as new biomarkers for EMT-like processes in lung cancer.