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Two new tremulane-type sesquiterpenes, irlactin L (1) and irlactin M (2) were isolated from cultures of the fungus Irpex lacteus, together with one known compound, 6-hydroxy-2,6-dimethyloct-7-enoic acid (3). Their structures were elucidated by spectroscopic data analysis. Compounds 1–2 were tested for their cytotoxicities against five human cancer cell lines and for their inhibitory activities against isozymes of 11β-hydroxysteroid dehydrogenases (11β-HSD). [ABSTRACT FROM AUTHOR]

Three new vibralactone derivatives, namely vibralactones U–W (1–3), together with vibralactone (4), have been isolated from cultures of the basidiomycete Boreostereum vibrans. Their structures were determined on the basis of spectroscopic methods and literature data. All compounds showed no activities to five human cancer cell lines. [ABSTRACT FROM AUTHOR]

Increasing evidence indicates that microRNAs (miRNAs) play an important role in various tumors by regulating downstream target genes and diverse signaling pathways. Herein, we confirmed miR-1265 expression in gastric cancer (GC) using the Cancer Genome Atlas (TCGA) database and assessed the level of miR-1265 expression in clinical specimens and cell lines. We found that miR-1265 expression was negatively correlated with tumor size. Further functional analysis revealed that miR-1265 suppresses cellular proliferation and autophagy while inducing apoptosis in GC cells. A luciferase reporter assay was used to identify an miR-1265 targeted gene, calcium binding protein 39 (CAB39), which is an essential upstream regulator in the AMPK-mTOR signaling pathway. Upregulation or downregulation of CAB39 expression reversed the effects of miR-1265 overexpression or inhibition, respectively. Notably, the knockdown of autophagy-related gene 12 (ATG12) impaired the effects of miR-1265 inhibition or CAB39 overexpression in GC. MiR-1265 also suppressed the growth of GC cells in vivo and that of human gastric organoids. Altogether, our results show that miR-1265 suppresses GC progression and oncogenic autophagy by reducing CAB39 expression and regulating the AMPK-mTOR signaling pathway. Therefore, miR-1265 may represent a potential therapeutic target for GC. [ABSTRACT FROM AUTHOR]

A new isopimarane diterpenoid, inonotolide D (1), was isolated from cultures of the basidiomycetes Inonotus sinensis. The new structure was elucidated on the basis of extensive spectroscopic studies. The compound was evaluated for its cytotoxicities against five human cancer cell lines. [ABSTRACT FROM AUTHOR]

A new tremulane sesquiterpene lactone, named irlactin K (1), was isolated from the culture broth of the basidiomycete Irpex lacteus. The structure of the new compound was established through extensive spectroscopic analyses. The compound was tested for its cytotoxicity against five human cancer cell lines and for its inhibitory activity against isozymes of 11β-hydroxysteroid dehydrogenases. [ABSTRACT FROM AUTHOR]

Five vibralactone derivatives containing γ , δ , ε -lactone cores, namely vibralactones X ( 1 ), Y ( 2 ), and Z 1 –Z 3 ( 3 – 5 ), together with the known vibralactone, have been isolated from cultures of the basidiomycete Boreostereum vibrans. Compounds 3 – 5 possessed a novel bis- γ -lactone group which was found in vibralactone derivatives for the first time. Compounds 3 and 5 exhibited moderate cytotoxicities to human cancer cell lines referring to that of cisplatin. [ABSTRACT FROM AUTHOR]

Background: In this work, we investigated the functional role of microRNA 137 (miR-137) in regulating osteosarcoma both in vitro and in vivo. Methods: Quantitative RT-PCR was used to examine the gene expressions of miR-137 in osteosarcoma cell lines and osteosarcoma tumors. 143B and Saos-2 cells were infected with lentivirus expressing miR-137 mimics (miR-137-mimic) to ectopically upregulate miR-137. In vitro cancer proliferation and migration were examined by MTT assay and transwell assay, respectively. Viral infected Saos-2 cells were also subcutaneously inoculated into null mice to evaluate the effect of miR-137 upregulation on in vivo tumor growth. The interaction between miR-137 and its downstream target, FXYD6, was evaluated by dual-luciferase reporter assay and quantitative real-time PCR. FXYD6 was then subsequently upregulated in osteosarcoma cells to evaluate its effect on miR-137 regulation in osteosarcoma. Results: We found that miR-137 was significantly downregulated in both osteosarcoma cell lines and osteosarcoma tumors. Lentiviral infection of miR-137-mimic upregulated miR-137 gene expression, reduced in vitro proliferation and migration and inhibited in vivo osteosarcoma tumor growth. FXYD6 was verified to be directly interacting with miR-137, and its subsequent upregulation reversed the inhibitory effect of miR-137 upregulation in osteosarcoma. Conclusion: We revealed novel functional role of miR-137 in osteosarcoma regulation, likely through FXYD6 binding. [ABSTRACT FROM AUTHOR]

Purpose: To investigate the effects of TRF2 depletion on radiosensitivity in both the telomerase-positive cell lines (A549) and alternative lengthening of telomere (ALT) cell lines (U2OS). Methods: X-ray irradiation was used to establish two radioresistant cancer models (A549R and U2OSR) from A549 and U2OS. Colony formation assay was applied to examine the radiosensitivity of radioresistant A549R and U2OSR cells and TRF2 low-expression cells. Real-time PCR and TeloTAGGG Telomerase PCR ELISA Kit were performed to examine telomere length and telomerase activity separately. γ-H2AX was detected by immunofluorescence to assess the radiation-induced DSBs. Results: Radioresistant cancer models were established, in which TRF2 was significantly over-expressed. Low expression of TRF2 protein could enhance the radiosensitivity and induce telomere length of A549 and U2OS cell shortening. In A549 cells with TRF2 down-regulated, the telomerase activity was inhibited, too. TRF2 deficiency increases γ-H2AX foci and fails to protect telomere from radiation. Conclusion: The data suggest that TRF2 is a radioresistant protein in A549 and U2OS cells, and could potentially be a target for radiosensitization of both telomerase-positive and ALT cells in radiotherapy. [ABSTRACT FROM AUTHOR]

Two new 3,4-seco-lanostane-type triterpenes, named as 15α-hydroxy-3,4-seco-lanosta-4(28),8,24-triene-3,21-dioic acid (1), 5α-hydroxy-3,4-seco-lanosta-4(28),8,24-triene-3,21-dioic acid 3-methyl ester (2), and one new lanostane triterpene 15α-acetoxylhydroxytrametenolic acid (3) together with a known one versisponic acid D (4) were isolated from the fruiting bodies ofLaetiporus sulphureusvar.miniatus. Their structures were determined on the basis of extensive spectroscopic methods and comparison with reported data. All four compounds were evaluated for their cytotoxicities against five human cancer cell lines; however, none exhibited inhibitory effects. [ABSTRACT FROM PUBLISHER]

Three new isopimarane diterpenoid lactones, named inonotolides A–C ( 1–3 ), were isolated from cultures of the basidiomycete Inonotus sinensis. Their structures were elucidated on the basis of extensive spectroscopic studies and the structure of inonotolide A ( 1 ) was confirmed by single-crystal X-ray crystallographic analysis. All compounds were evaluated for their cytotoxicities against five human cancer cell lines. [ABSTRACT FROM AUTHOR]

One new ergostane-type steroid, (12β,15β,22R,23S,24S)-22,25-epoxy-12,15,23-trihydroxyergost-4,6,8(14)-trien-3-one (1), three new phthalide derivatives, 5-(2′,3′-epoxy-3′,3′-dimethylpropoxy)-7-methoxy-6-methylphthalide (2), (2′)-(Z)-5-(3′-hydroxymethyl-3′-methylallyloxy)-7-methoxy-6-methylphthalide (3), and 5-(3′,3′-dimethylallyloxy)-7-hydroxy-6-methylphthalide (4), along with one known phthalide derivative, 5-(3′,3′-dimethylallyloxy)-7-methoxy-6-methylphthalide (5), were isolated from cultures of the basidiomyceteAlbatrellus confluens. The structures of the new compounds were established on the basis of extensive spectroscopic data (IR, MS, 1D, and 2D NMR) analyses. All compounds were evaluated for their cytotoxic activities on five tumor cell lines. [ABSTRACT FROM PUBLISHER]

Three new sesquiterpenoids (1–3), along with four known compounds (4–7), were isolated from cultures of the fungus Conocybe siliginea. The structures of new compounds were elucidated using spectroscopic methods. The known compounds were identified by comparing their spectroscopic data with those reported in the literature. All new compounds were evaluated for cytotoxicity against five human cancer cell lines, but none of them possesses significant activity (IC50>40μM). [ABSTRACT FROM AUTHOR]

Purpose: Drug resistance is not only one of the major obstacles to treatment but also a poor prognosis in advanced non-small cell lung cancer (NSCLC) patients. The aim of this study was to evaluate the predictive value of APE1, BRCA1, ERCC1 and TUBB3 in advanced NSCLC patients who received platinum-paclitaxel treatment. Methods: One hundred and thirty-six advanced NSCLC patients, who were treated with first-line platinum-paclitaxel chemotherapy, were enrolled in this study. The protein expression levels of APE1, BRCA1, ERCC1 and TUBB3 were assessed by immunohistochemistry and analyzed for the association with response to chemotherapy and progression-free survival (PFS) and overall survival (OS). Results: Patients with negative expression of APE1, ERCC1 or TUBB3 benefited from platinum plus paclitaxel regimen chemotherapy. ERCC1-negative patients had better PFS ( P = 0.016) and OS ( P = 0.030) compared with positive patients. Similarly, the APE1-negative patients showed better PFS ( P = 0.004) and longer OS though statistically insignificant. Multivariate analysis showed that APE1 and ERCC1 were independent predictor for PFS (HR 2.07; P = 0.004 and HR 1.66; P = 0.016) and OS (HR 1.99; P = 0.008 and HR 1.64; P = 0.040). Moreover, patients with both APE1- and ERCC1-negative or both APE1- and TUBB3-negative tumors had significantly higher response rate, longer median PFS and OS following treatment with platinum and paclitaxel ( P < 0.05). Conclusion: The data indicate that APE1, ERCC1 and TUBB3 could be a useful biomarker to predict clinical outcome in patients with advanced NSCLC receiving first-line platinum-paclitaxel chemotherapy. [ABSTRACT FROM AUTHOR]

Osterix, a zinc-finger-containing transcription factor, is required for osteoblast differentiation and bone formation. Osterix is also expressed in dental mesenchymal cells of the tooth germ. However, transcriptional regulation by Osterix in tooth development is not clear. Genetic studies in osteogenesis place Osterix downstream of Runx2 (Runt-related 2). The expression of Osterix in odontoblasts overlaps with Runx3 during terminal differentiation in vivo. Runx3 down-regulates Osterix expression in mouse DPCs (dental pulp cells). Therefore the regulatory role of Runx3 on Osterix expression in tooth development was investigated. Enforced expression of Runx3 down-regulated the activity of the Osterix promoter in the human embryonic kidney 293 cell line. When the Runx3 responsive element on the Osterix promoter, located at −713 to −707 bp (site 3, AGTGGTT) relative to the cap site, was mutated, this down-regulation was abrogated. Furthermore, electrophoretic mobility-shift assay and chromatin immunoprecipitation assays in mouse DPCs demonstrated direct functional binding of Runx3 to the Osterix promoter. These results demonstrate the transcriptional regulation of Osterix expression by Runx3 during differentiation of dental pulp cells into odontoblasts during tooth development. [ABSTRACT FROM AUTHOR]

[Display omitted] • Novel 2,4-diarylaminopyrimidine analogues bearing 1 H -benzo[ d ]imidazol motif as potent ALK inhibitors; • H-11 inhibited ALK-addicted cancer cells with the IC 50 values superior to the reference ceritinib; • H-11 possessed remarkable potency against ALKWT, ALKL1196M and ALKG1202R kinases with IC 50 values below 5.7 nM; • Molecular docking study supported that H-11 was a promising ALK inhibitor for mutations. To address drug resistance caused by ALK kinase mutations, a series of novel 2,4-diarylaminopyrimidine (DAAP) analogues were designed by incorporating 1 H -benzo[ d ]imidazol motif onto the maternal framework. All compounds were efficiently synthesized and antiproliferative activities against Karpas299, H2228 and A549 cell lines were evaluated by MTT assay. Delightly, the most promising derivative H-11 was detected with IC 50 values of 0.016 μM and 0.099 μM against ALK- positive Karpas299 and H2228 cells. Meanwhile, H-11 displayed encouraging enzymatic inhibitory potency with IC 50 values of 2.7 nM, 3.8 nM and 5.7 nM toward ALKWT, ALKL1196M and ALKG1202R, respectively. Ultimately, the binding modes of optimal H-11 with ALK wild-type and mutants were ideally established which further confirmed the structural basis in accordance with the SARs analysis. [ABSTRACT FROM AUTHOR]

Acrylamide (ACR) is a water-soluble chemical applied in industrial and laboratory processes. The neurotoxicity induced by acrylamide involves both peripheral and central nervous system. Hence, there is a growing urgency to investigate the mechanisms of acrylamide-induced neurotoxicity and search novel therapeutic target for the nerve repair. The effects of ACR on the proliferation, reactive oxygen species (ROS) and iron production of dorsal root ganglia (DRG) neurons and Schwann cells were determined. 5-Ethynyl-2′-deoxyuridine (EDU) staining and transwell assay were applied to detect the proliferation and migration capacity of DRG cells. Ferrostatin-1 (Fer-1) was used to suppress ferroptosis induced by ACR. RT-PCR analysis was performed to examine the expression of neurotrophic factors including brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), vascular endothelial growth factor (VEGF) and glial cell line-derived neurotrophic factor (GDNF). Moreover, Iron, ROS, malondialdehyde (MDA) and glutathione (GSH) contents were measured to reveal the regulation of ferroptosis in ACR-related nerve injury. ACR inhibited the proliferation and migration of DRG neurons and the supplementation of Fer-1 reversed the effects induced by ACR. Besides, the treatment of Fer-1 effectively increased the expression of NGF, BDNF, VEGF and GDNF. Furthermore, ACR increased the iron level, MDA and ROS contents while inhibited the level of GSH. It was unveiled that ACR attenuated the proliferation, migration and neuron repair of DRG neurons through regulating ferroptosis. The modulation of ferroptosis might be a promising therapeutic strategy and provide references for future treatment of acrylamide-induced nerve damage. [ABSTRACT FROM AUTHOR]

Abstract: Two novel iridoids triohimas A (1) and C (3) with an unusual δ-lactone-containing skeleton were isolated from Triosteum himalayanum Wall. Their structures were determined by NMR spectroscopic analyses and X-ray crystallography. The absolute configuration was established by computational methods. They were also tested for the in vitro cytotoxicity against L1210 cell line. [Copyright &y& Elsevier]

Five new cytochalasans (1 – 5) were isolated from the rice fermentation of fungus Xylaria longipes , along with seven known compounds cytochalasin P (6), cytochalasin D (7), zygosporin D (8), 7- O -acetylcytochalasin D (9), cytochalasin C (10), 6,7-dihydro-7-oxo-cytochalasin C (11), and 6,7-dihydro-7-oxo-deacetylcytochalasin C (12). Their structures and absolute configurations were determined by extensive experimental spectroscopic methods as well as ECD calculation and GIAO 13C NMR calculation. The cytotoxicity of obtained compounds (1 − 12) was evaluated against human cancer cell lines HL-60, A549, SMMC-7721, MCF-7, and SW480. Compounds 6 – 8 , 11 , and 12 showed cytotoxicity with IC 50 value ranging from 4.17–37.18 μM. Unlabelled Image [ABSTRACT FROM AUTHOR]

Nine new 19,20-epoxycytochalasans (1 – 9) were isolated from the rice fermentation extracts of endophytic fungus Xylaria cf. curta , along with four known compounds 19,20-epoxycytochalasin C (10), 18-desoxy-19,20-epoxycytochalasin C (11), 19,20-epoxycytochalasin D (12) and 5,6-dihydro-7-oxo-19,20-epoxycytochalasin C (13). Their structures and absolute configurations were determined by 1D and 2D NMR, HRESIMS, X-ray diffraction and ECD calculation. The cytotoxicity of obtained compounds (1 − 13) was evaluated against human cancer cell lines HL-60, A549, SMMC-7721, MCF-7, and SW480. Remarkably, compound 10 showed significant specific cytotoxicity against HL-60 cell lines with IC 50 value of 1.11 μM. Unlabelled Image [ABSTRACT FROM AUTHOR]

Breast cancer is considered the most prevalent malignancy due to its high incidence rate, recurrence, and metastasis in women that makes it one of the deadliest cancers. The current study aimed to predict the genes associated with the recurrence and metastasis of breast cancer and to validate their effect on MDA-MB-231 cells. Through the bioinformatics analysis, the transcription factor 7 cofactor (MLLT11) as the target gene was obtained. MLLT11-specific siRNA was synthesized and transfected into MDA-MB-231 cells. The results demonstrated that the siRNA significantly reduced the MLLT11 mRNA levels. Moreover, cell migration and invasion, as well as the protein levels of phosphatidylinositol 3-kinase (PI3K), AKT, matrix metalloproteinase (MMP) 2, and MMP9, were significantly lower in the groups treated with siRNA while the apoptosis was augmented. Collectively, MLLT11 siRNA elicited ameliorative properties on breast cancer cells, possibly via the inhibition of the PI3K/AKT signaling pathway. [ABSTRACT FROM AUTHOR]

Two new compounds, daedatrin K (1) and 2-hydroxy-1-(5-(hydroxymethyl)furan-2-yl)propan-1-one (2), were isolated from cultures of the basidiomycetes Daedaleopsis tricolor. The new structures were elucidated on the basis of extensive spectroscopic methods. At the same time, two compounds were tested for their cytotoxicities against five human cancer cell lines. [ABSTRACT FROM AUTHOR]

(–)-5-Methylmellein (1) and its new dimer (2) were isolated from cultures of the basidiomycete Inonotus sinensis. Their structures were elucidated on the basis of extensive spectroscopic methods including UV, IR, HR-EI-MS, 1D NMR and 2D NMR. The structure of Compound 2 was determined by single-crystal X-ray crystallographic analysis. Compound 2 was tested for the cytotoxicities against five human cancer cell lines. [ABSTRACT FROM AUTHOR]

A series of oxygenated yohimbane alkaloids, including three new compounds, ophiorrhines H-J (1 – 3), and seven known compounds, were isolated from the aerial parts of Ophiorrhiza japonica. The structures with absolute configurations were elucidated by extensive MS and NMR spectroscopic methods, as well as the single crystal X-ray diffraction and ECD calculations. Ophiorrhines H (1) and I (2) represent key oxygenated intermediates in the formation of aromatic ring E in the demethoxycarbonyl-3,14-dihydrogambirtannine (10). Ophiorrhine J (3) is a highly oxidized yohimbane derivative with the planar superconjugated system. The cytotoxic activities of all alkaloids against five human cancer cell lines were evaluated. [Display omitted] [ABSTRACT FROM AUTHOR]

Purpose: Mesenchymal stromal cells (MSCs) hold promise in the treatment of liver disease. However, short survival time of MSCs after intrahepatic transplantation limits their value; therefore, understanding the basis of MSCs survival and rejection may increase their utility. This study was aimed at determining the role of intrahepatic natural killer (NK) cells on MSCs survival and their retention in the liver shortly after transplant.Procedures: Human MSCs were labeled with the Luc2-mKate2 dual-fusion reporter gene (MSCs-R), and the residence time and survival of MSCs-R xenografts after intrahepatic transplantation were evaluated by in vivo bioluminescence imaging (BLI). Coculture of MSCs and NK cells was performed to assess cytotoxicity. To evaluate the role of NK cells in rejection of the xenografted cells, the fates of transplanted MSCs-R were then assessed in vivo by BLI after activation of intrahepatic NK cells.Results: We observed a linear correlation between luciferase activity from live MSCs-R and cell number in vitro (R 2 = 0.9956). In vivo, we observed a gradual decline in bioluminescent signals from transplanted MSCs-R over a region corresponding to the liver in both the control group and the NK-activated group. However, the survival time and retention of intrahepatic MSCs-R decreased more rapidly in the NK-activated group of mice compared to the control group. This indicated that activated NK cells accelerate the elimination of transplanted MSCs. Also, we found that the number of hepatic NK cells and the expression of NK activation markers significantly increased after intrahepatic delivery of MSCs. This suggested that resident NK cells, in a resting state, were activated by intrahepatic transplantation of human MSCs. Taken together, the data suggests that activated hepatic NK cells mediate, in part, rejection of the MSCs xenografts. Cytotoxicity assays showed that activated NK cells may inhibit the proliferation of MSCs and, to a certain extent, induce MSCs death.Conclusion: Human MSCs could be followed dynamically in vivo by BLI, and the role of murine hepatic NK cells, especially activated NK cells, could be inferred from the loss of signals from MSCs. This finding may have practical clinical implications in MSCs transplantation in treating liver disease. [ABSTRACT FROM AUTHOR]

Improving productivity to reduce the cost of biologics manufacturing and ensure that therapeutics can reach more patients remains a major challenge faced by the biopharmaceutical industry. Chinese hamster ovary (CHO) cell lines are commonly prepared for biomanufacturing by single cell cloning post-transfection and recovery, followed by lead clone screening, generation of a research cell bank (RCB), cell culture process development, and manufacturing of a master cell bank (MCB) to be used in early phase clinical manufacturing. In this study, it was found that an additional round of cloning and clone selection from an established monoclonal RCB or MCB (i.e., re-cloning) significantly improved titer for multiple late phase monoclonal antibody upstream processes. Quality attributes remained comparable between the processes using the parental clones and the re-clones. For two CHO cells expressing different antibodies, the re-clone performance was successfully scaled up at 500-L or at 2000-L bioreactor scales, demonstrating for the first time that the re-clone is suitable for late phase and commercial manufacturing processes for improvement of titer while maintaining comparable product quality to the early phase process. [ABSTRACT FROM AUTHOR]

Two new sesquiterpenoids (phellinuins H and I), together with five known compounds, were isolated from cultures of mushroom Phellinus sp. Their structures were elucidated based on comparison of nuclear magnetic resonance and MS data and those reported in the literature. All of these compounds were tested for cytotoxicity against five cancer cell lines (HL-60, SMMC-721, A-549, MCF-7, and SW-480). [ABSTRACT FROM AUTHOR]

Background: Increasing evidence has demonstrated that long noncoding RNAs (lncRNAs) have regulatory functions in hepatocellular carcinoma (HCC). The link between lincSCRG1 and HCC remains unclear. Methods: To explore the lincSCRG1 regulation axis, bioinformatics, RIP and luciferase reporter assay were performed. The expressions of lincSCRG1-miR26a-SKP2 were detected in HCC tissues and cell lines through qPCR and western blot. The functions of HCC cells were investigated through in vitro assays (MTT, colony formation, transwell and flow cytometry) and the inner effect of lincSCRG1-miR26a in vivo was evaluated by xenografts and liver metatstatic nude mice models. Results: LincSCRG1 was found to be strongly elevated in human HCC tissues and cell lines. MiR26a and S phase kinase-related protein 2 (SKP2) were predicted as the target miRNA for lincSCRG1 and the target gene for miR26a with direct binding sites, respectively. LincSCRG1 was verified as a competing endogenous RNA (ceRNA) via negative regulation of miR26a and derepression of SKP2 in HCC cells. Both overexpression of lincSCRG1 (ov-lincSCRG1) and inhibition of miR26a (in-miR26a) obviously stimulated cellular viability, colony formation, migration and proliferation of S phase cells and also significantly increased the protein levels of cyclinD1, CDK4, MMP2/3/9, Vimentin, and N-cadherin or inhibited the protein level of E-cadherin of HCC cells, while knockdown of lincSCRG1 (sh-lincSCRG1) and upregulation of miR26a (mi-miR26a) had the opposite effects on HCC cells. Cotransfection of in-miR26a or overexpression of SKP2 (ov-SKP2) with sh-lincSCRG1 could rescue the anticancer functions of sh-lincSCRG1, including suppressing proliferation and migration of HCC cells. Additionally, sh-lincSCRG1 could effectively inhibit the growth of subcutaneous xenograft tumours and lung metastasis, while the anticancer effect of sh-lincSCRG1 could be reversed by cotransfection of in-miR26a. Conclusions: LincSCRG1 acts as a ceRNA of miR26a to restrict its ability to derepress SKP2, thereby inducing the proliferation and migration of HCC cells in vitro and in vivo. Depletion of lincSCRG1 could be used as a potential therapeutic approach in HCC. [ABSTRACT FROM AUTHOR]

Background: A recent study reported a novel long non-coding RNA (lncRNA) E2F-mediated cell proliferation enhancing lncRNA (EPEL, human chromosome 4, intergenic region) plays an oncogenic role in lung cancer.Aims: We aimed to investigate the role of lncRNA EPEL in gastric cancer.Methods: Gene expression was analyzed by RT-qPCR and western blot. Survival analysis was performed by comparing survival curves. Cell proliferation, migration, and invasion were analyzed by CCK-8 and Transwell assays.Results: We found that lncRNA EPEL and Runt-related transcription factor 2 (RUNX2) were both upregulated in gastric cancer. EPEL and RUNX2 were positively correlated in tumor. Patients with high expression level of lncRNA EPEL showed poor survival. LncRNA EPEL and RUNX2 overexpression promoted, while lncRNA EPEL siRNA silencing inhibited the migration, proliferation, and invasion of gastric cancers. In addition, RUNX2 overexpression completely rescued the inhibited cancer cell migration, proliferation, and invasion caused by lncRNA EPEL siRNA silencing. Consistently, EPEL overexpression resulted in upregulated RUNX2 expression, while RUNX2 overexpression did not affect lncRNA EPEL expression.Conclusions: Therefore, lncRNA EPEL may regulate cancer cell behaviors and affect prognosis of gastric cancer by interacting with RUNX2. [ABSTRACT FROM AUTHOR]

Increasing cell culture productivity of recombinant proteins via process improvements is the primary focus for research groups within biologics manufacturing. Any recommendations to improve a manufacturing process obviously must be effective, but also be robust, scalable, and with product quality comparable to the original process. In this study, we report that three different GS−/− CHO cell lines developed in media containing a standard concentration of the selection agent methionine sulfoximine (MSX), but then exposed to increased MSX concentrations during seed train expansion, achieved titer increases of 10–19%. This result was observed in processes already considerably optimized. Expanding the cells with a higher MSX concentration improved cell line production stability with increased culture age. Production cultures in 500‐L and 1000‐L bioreactors replicated laboratory results using 5‐L bioreactors, demonstrating process robustness and scalability. Furthermore, product quality attributes of the final drug substance using the higher MSX process were comparable with those from cells expanded in media with the standard selection MSX concentration. Subsequent mechanistic investigations confirmed that the cells were not altered at the genetic level in terms of integration profiles or gene copy number, nor transcriptional levels of glutamine synthetase, heavy chain, or light chain genes. This study provides an effective and applicable strategy to improve the productivity of therapeutic proteins for biologics manufacturing. [ABSTRACT FROM AUTHOR]

Four new alkaloids, melodinines W 1 –W 4 (1 – 4), together with twenty one known alkaloids (5 – 25) were isolated from Melodinus henryi. The structures with absolute configurations were elucidated by extensive MS and NMR spectroscopic methods, as well as the single crystal X-ray diffraction and ECD calculations. All compounds were evaluated for their cytotoxicities to five human cancer cell lines. Many compounds showed certain cytotoxicities to five human cancer cell lines with an IC 50 range of 1.4–29.4 μM. Unlabelled Image [ABSTRACT FROM AUTHOR]

Ten previously undescribed triterpenoid congeners, namely irpeksolactins A–J, together with eighteen known ones, were isolated from the fruiting bodies of the rainforest-dwelling medicinal fungus Irpex lacteus. The structures of all the isolates were characterized by extensive spectroscopic approaches, including 1D & 2D NMR and MS spectroscopic methods. Irpeksolactin J displayed selective and weak cytotoxicity against the human lung cancer cell line A549 and the human hepatocellular carcinoma cell line SMMC-7721. Image 1 • Twenty-eight triterpenes are reported from Irpex lacteus. • Irpeksolactin A is featured by a rare 19 (10. → 5) abeo -eburicane skeleton. • Irpeksolactins F and G are featured by a rare 5,8-peroxy bridge. • Irpeksolactin J is featured by a rare 16,24-epoxy group. • Irpeksolactin J shows weak cytotoxicity. [ABSTRACT FROM AUTHOR]

Abstract Two undescribed piperidine racemates, (±)-caulophines A and B (1 and 2), a new N -containing xanthone derivative (3), together with six known piperidines, were isolated from the roots of Caulophyllum robustum Maxim. Their structures were determined by extensive spectroscopic techniques. Compounds 3 and 7 exhibited weak cytotoxicities against human palace cancer hela cell line with inhibitory rates of 32.2% and 39.7%, respectively, at the concentration of 40 μM. Graphical abstract Unlabelled Image [ABSTRACT FROM AUTHOR]

Objectives: To investigate the effect of kallikrein-related peptidase KLK1 on azoospermic mice induced by busulfan and mouse spermatogonial stem cell.Methods: Mice were treated with a single intraperitoneal injection of busulfan, and 4 weeks later, they received a daily intraperitoneal injection of KLK1 at different doses for another 4 weeks. Eight weeks after the busulfan treatment, all mice were sacrificed and their testes were collected for histological evaluation, immunostaining and protein extraction. In vitro, immortalized mouse spermatogonial stem cells, namely C18-4 cells, were treated with KLK1 for proliferation assays.Results: Histological evaluation of testes, epididymis and epididymal fluid showed that KLK1-treated mice had better spermatogenesis than the control group. Immunostaining showed that tissue samples from testes of KLK1-treated mice had more PLZF- and SCP3-positive cells per seminiferous tubule as well as more PNA-positive cells in the seminiferous tubules. Western blots revealed higher expression levels of PCNA in KLK1-treated mice than in control mice. C18-4 cells treated with KLK1 had a higher proliferation rate and higher expression levels of PCNA, Cyclin A and Cyclin E, and the level of phosphorylated ERK2 were increased after KLK1 treatment.Conclusion: Collectively, KLK1 can improve spermatogenesis in azoospermic mice, and KLK1 can promote the proliferation of mouse spermatogonial stem cells via activating ERK1/2 and cell cycle proteins Cyclin A and Cyclin E. This study could offer novel approach and provide new targets for the treatment of azoospermia. [ABSTRACT FROM AUTHOR]

A highly degraded seco -triterpene derivative, panisuffrutin A (1), from Paeonia suffruticosa var. papaveracea (Andr.) Kerner. • A highly degraded seco -triterpene derivative (1) has been isolated from Paeonia suffruticosa var. papaveracea (Andr.) Kerner. • The structure with absolute configuration was determined by NMR methods and ECD calculations. • Compound 1 showed certain cytotoxicity to human Hela cancer cell line. Panisuffrutin A (1), a highly degraded seco -triterpene derivative, together with the known palbinone, has been isolated from the whole plant Paeonia suffruticosa var. papaveracea (Andr.) Kerner. The structure with absolute configuration of 1 was determined via comprehensive NMR and MS analyses, as well as NMR and ECD calculations. A plausible biosynthetic pathway for 1 was proposed. Compound 1 showed weak cytotoxicity against Hela cancer cell line with an IC 50 value of 26.2 μM, while palbinone exhibited a moderate inhibition on NO production with an IC 50 of 18.3 μM. [ABSTRACT FROM AUTHOR]

Chemical investigation on the cultures ofPhellinus tuberculosusandLaetiporus sulphureuslead to the isolation of two new illudin-type sesquiterpenoids (phellinuin J and sulphureuine A). Their structures were elucidated by 1D, 2D NMR and MS spectroscopic data. These compounds were purposely evaluated for their cytotoxicity against HL-60, SMMC-7721, A549, MCF-7, and SW480 cell lines. [ABSTRACT FROM PUBLISHER]

Two new triterpenoids, (24E)-9α,11α-epoxy-3β-hydroxylanosta-7,24-dien-26-al (1) and (22Z,24Z)-13-hydroxy-3-oxo-14(13 → 12)abeo-lanosta-8,22,24-trien-26,23-olide (2) were isolated from dried fruiting bodies of fungusGanoderma lucidum. The structures of these two new compounds were elucidated on the basis of extensive spectroscopic analyses. Compound1possessed a lanostane skeleton, while compound2was based on a rare 14 (13 → 12)abeo-lanostaneskeleton with a 26,23-olide moiety. Both of them were evaluated for their antifungal and cytotoxic activities. Neither of them displayed obvious inhibition onCandida albicansand five human cancer cell lines. [ABSTRACT FROM PUBLISHER]

Three new bisabolane sesquiterpenoids, inonotic acid A (1), 3-O-formyl inonotic acid A (2), inonotic acid B (3), and one new drimane sesquiterpenoid 3α,6β-dihydroxycinnamolide (4), were isolated from the fruiting bodies of mushroomInonotus rickii. Their structures were elucidated by means of extensive spectroscopic methods. Compound4had moderate inhibitory activity on human colon cancer SW480 (IC50 = 20.4 μmol). [ABSTRACT FROM PUBLISHER]

Abstract: Low levels of alanine to serine sequence variants were identified in an IgG4 monoclonal antibody by ultra/high performance liquid chromatography and tandem mass spectrometry. The levels of the identified sequence variants A183S and A152S, both in the light chain, have been determined to be 7.8–9.9% and 0.5–0.6%, by extracted ion currents of the tryptic peptides L16 and L14, respectively. The A183S variant was confirmed through tryptic map spiking experiments using synthetic peptide, SDYEK, which incorporated Ser at the position of native Ala in the tryptic peptide L16. Both mutations were also observed by endoproteinase Asp-N peptide mapping. The variant level of A183S was also quantified by LC–UV with detection at 280nm and fluorescence detection of tyrosine residues on the tryptic peptides. The results from LC–MS, UV, and fluorescence detection are in close agreement with each other. The levels of the sequence variants are comparable among the antibody samples manufactured at different scales as well as locations, indicating that the variants’ levels are not affected by manufacture scale or locations. DNA sequencing of the master cell bank revealed the presence of mixed bases at position 183 encoding both wild and mutated populations, whereas bases encoding the minor sequence variant at position 152 were not detected. The root cause for A152S mutation is not yet clearly understood at this moment. [Copyright &y& Elsevier]

Abstract: A series of phenanthroindolizidine and phenanthroquinolizidine alkaloids and their 14-amino-derivatives (1–44) were prepared and systematically evaluated for their anti-tumor activities against A549 and HL60 cell lines. The bioassay results showed that most of these alkaloids possess good anti-tumor activities. Especially, compounds 15, 22, 28, 33–36, 40 and 42 displayed low nanomolar or subnanomolar levels of anti-tumor activity. The configuration of (13aS,14S)-14-hydroxyphenanthroindolizidines and (14aR,15R)-15-hydroxyphenanthroquinolizidines was confirmed to be optimal. 14-Amino-phenanthroindolizidines with increased polarity possess good anti-tumor activity, especially for compounds 26 and 28. Most of the phenanthroquinolizidine alkaloids exhibited higher anti-tumor activity than that of phenanthroindolizidine alkaloids. Our present study provides fundamental support for development and optimization of phenanthroindolizidine and phenanthroquinolizidine alkaloids as potential anti-tumor drugs. [Copyright &y& Elsevier]

Gingival fibroblast cell lines were derived from Sorsby''s fundus dystrophy (SFD) patients carrying the S181C TIMP3 and the E139X TIMP3 mutations. These cell lines were grown in culture to study expression of the wild-type and mutant tissue inhibitor of metalloproteinase 3 (TIMP3) alleles from a normal diploid cell type. Firstly, patient cells were found to co-express the wild-type and mutant TIMP3 alleles, S181C TIMP3 or E139X TIMP3, at the mRNA level using restriction fragment length polymorphism (RFLP) analysis. A SpeI RFLP for E139X TIMP3 is described. Low levels of endogenous TIMP3 protein expression were elevated using the natural polysaccharide calcium pentosan polysulfate (CaPPs) in combination with the cytokine IL-1α. Immunoblotting detected protein expression from both wild-type and mutant alleles, S181C TIMP3 or E139X TIMP3. S181C TIMP3 from these cells was found to dimerise and retain MMP2 inhibitory activity. To facilitate studies of the E139X TIMP3 protein, the allele was expressed using HighFive insect cells. In this cell type, the E139X TIMP3 was synthesised as a mixture of monomer and dimer. Both monomeric and dimeric E139X TIMP3 protein retained MMP2 inhibitory activity in gelatin zymography. Expression of mutant E139X or S181C TIMP3 protein from a normal diploid patient-derived fibroblast cell had no effect on either MMP2 or MMP9 expression or activation whilst transcribed from their normal promoter context. [Copyright &y& Elsevier]

The chemical constituents and their biological activities of the mushroom Pyropolyporus fomentarius were investigated in this study. Two previously undescribed pentacyclic lupane-type triterpenes, 3-formyloxybetulin and 3-formyloxybetulinic acid, two rare degraded ergosterols, pyropolincisterols A and B, along with ten known triterpenoids and four known ergosterols were isolated from the fruiting bodies of P. fomentarius. Their chemical structures were determined using a combination of spectroscopic analysis. Nine compounds exhibited certain cytotoxicities to human cancer cell lines, while polyporenic acid showed significant cytotoxicities to SMMC-7721 and A-549 with IC 50 values less than 10 μ M. Four compounds showed inhibitory activities against nitric oxide (NO) production in LPS-activated RAW264.7 macrophages with IC 50 values of 36.3, 25.1, 21.4, and 34.2 μ M, respectively. The results of this assessment suggested that the lanostane triterpenoids and ergosterols in fruiting bodies of P. fomentarius played key roles in its folk usages. Triterpenoids and ergosterols from mushroom Pyropolyporus fomentarius with cytotoxic and NO production inhibitory activities. Image 1 • A total of 18 compounds have been obtained from mushroom Pyropolyporus fomentarius. • Two compounds were rare carbon-degraded ergosterol derivatives. • Many compounds possess cytotoxicities and NO production inhibitory activities. [ABSTRACT FROM AUTHOR]

Background: Increasing evidence has demonstrated that long noncoding RNAs (lncRNAs) have regulatory functions in hepatocellular carcinoma (HCC). The link between lincSCRG1 and HCC remains unclear. Methods: To explore the lincSCRG1 regulation axis, bioinformatics, RIP and luciferase reporter assay were performed. The expressions of lincSCRG1-miR26a-SKP2 were detected in HCC tissues and cell lines through qPCR and western blot. The functions of HCC cells were investigated through in vitro assays (MTT, colony formation, transwell and flow cytometry) and the inner effect of lincSCRG1-miR26a in vivo was evaluated by xenografts and liver metatstatic nude mice models. Results: LincSCRG1 was found to be strongly elevated in human HCC tissues and cell lines. MiR26a and S phase kinase-related protein 2 (SKP2) were predicted as the target miRNA for lincSCRG1 and the target gene for miR26a with direct binding sites, respectively. LincSCRG1 was verified as a competing endogenous RNA (ceRNA) via negative regulation of miR26a and derepression of SKP2 in HCC cells. Both overexpression of lincSCRG1 (ov-lincSCRG1) and inhibition of miR26a (in-miR26a) obviously stimulated cellular viability, colony formation, migration and proliferation of S phase cells and also significantly increased the protein levels of cyclinD1, CDK4, MMP2/3/9, Vimentin, and N-cadherin or inhibited the protein level of E-cadherin of HCC cells, while knockdown of lincSCRG1 (sh-lincSCRG1) and upregulation of miR26a (mi-miR26a) had the opposite effects on HCC cells. Cotransfection of in-miR26a or overexpression of SKP2 (ov-SKP2) with sh-lincSCRG1 could rescue the anticancer functions of sh-lincSCRG1, including suppressing proliferation and migration of HCC cells. Additionally, sh-lincSCRG1 could effectively inhibit the growth of subcutaneous xenograft tumours and lung metastasis, while the anticancer effect of sh-lincSCRG1 could be reversed by cotransfection of in-miR26a. Conclusions: LincSCRG1 acts as a ceRNA of miR26a to restrict its ability to derepress SKP2, thereby inducing the proliferation and migration of HCC cells in vitro and in vivo. Depletion of lincSCRG1 could be used as a potential therapeutic approach in HCC. [ABSTRACT FROM AUTHOR]

• Transcriptomics and metabolomics were analyzed on an unstable industrial cell line. • Survival mechanism maintained robust cell growth. • Lysosomal pathway, redox balancing and lipid oxidation were enhanced over time. • DNA damage recognition and repair were disconnected in the aged cells. Cell line instability can pose a challenge from both regulatory and business needs perspectives. The regulatory guidelines of International Conference on Harmonization require manufacturers to evaluate the cell substrate with respect to the consistent production of the intended product and perform tests of stability during cell cultivation. In an effort to develop strategies for stabilizing cell lines, we applied RNA-seq-based transcriptomics and NMR and LCMS-based metabolomics to analyze an industrial monoclonal antibody-producing Chinese hamster ovary (CHO) cell line that was genetically unstable. The cells adapted well to environmental and nutritional changes during passaging and maintained robust cell growth, however, the omics data demonstrated that lysosome pathway, redox balancing, and lipid peroxidation and beta-oxidation were enhanced in aged cells. The genes involved in lysosome pathway activation up-regulated cellular recycling and scavenging activities, and reactive oxidative species production. While the genes associated with DNA damage were up-regulated and cell cycle was correspondingly inhibited, surprisingly those related to DNA repair machinery were down-regulated over time. This previously unrecognized disconnect between DNA damage recognition and the needed DNA repair highlights potential directions to stabilize cell lines by formulating inoculum media with increased antioxidant capacity or by maintaining a robust DNA repair system in CHO cells. Both directions require further study. [ABSTRACT FROM AUTHOR]

Two undescribed triterpenes, tricholimbrins A and B, three undescribed steroids, tricholimbrins C‒E, one undescribed 4-chromanone derivative, along with 27 known compounds were isolated from fruiting bodies of the mushroom Tricholoma imbricatum. Tricholimbrins A and B are two polycyclic triterpenoids with a carbon degradation, while tricholimbrin C is a ring-rearranged steroid containing an aromatic moiety that might be derived from an ergosterol. Isocyathisterol, 3 β ,15 α -dihydroxyl-(22 E ,24 R)-ergosta-5,8(14),22-trien-7-one, demethylincisterol A 3 , and volemolide showed cytotoxicities to six human cancer cell lines. 3 β -Hydroxyl-(22 E ,24 R)-ergosta-5,8,22-trien-7,15-dione and 3 β -hydroxyl-(22 E ,24 R)-ergosta-5,8,22-trien-7-one showed preferable cytotoxicities against HL-60 while chaxine C and volemolide showed preferable cytotoxicities against A-549, with IC 50 values less than 10 μ M. Triterpenoids and ergosterols from fruiting bodies of Tricholoma imbricatum. Image 1 • A total of 33 compounds have been obtained from mushroom Tricholoma imbricatum. • Compounds 1–3 are rare natural triterpenoids or ergosterol derivatives. • Many compounds possess certain cytotoxicities to human cancer cell lines. [ABSTRACT FROM AUTHOR]

• A genome-scale CHO model was employed to analyze the metabolism of CHO cells. • Flux analysis results from genome-scale model were compared with transcriptomics analysis. • In silico simulations using genome-scale model were performed for feed optimization. • This is the first study to apply genome-scale CHO model to improve the IgG production. Effective bioprocess development using Chinese hamster ovary (CHO) cells as hosts is hampered by the limited understanding of cellular metabolism under process conditions in bioreactors. Systematic tools such as genome-scale models have been developed, but their value has not been satisfactorily demonstrated and exploited for process development. In this study, we proposed a method using a genome-scale model to analyze existing process studies for bioprocess optimization. First, we used existing industrial CHO cell culture experiments to systematically gain metabolic insights for bioprocess development. Two fed-batch cultures, using the same cell line and process, resulted in different titers by supplementing two different types of feed media. A genome-scale model was applied to calculate fluxomics (i.e., intracellular fluxes) from extracellular metabolomics and then the metabolic differences were further analyzed through pathway analysis between these two cell culture conditions. Transcriptomics data from RNA-Seq were employed at this point for comparison and found to be consistent in pathway analysis with the flux analysis results. At the second stage, we developed a modeling-based approach for media optimization to increase antibody production based on the understanding from the first stage. The new design was tested in silico using a genome-scale model and then verified experimentally, confirming the applicability of this modeling-based approach for bioprocess optimization. The framework proposed in this study can maximize the utilization of existing process studies and minimize the time consumed for empirical work in developing new processes. [ABSTRACT FROM AUTHOR]

Three depsidones boremexins A–C (1 – 3), two diaryl ethers boremexins D (4) and E (5), together with four known compounds were obtained from cultures of potato endophytic fungus Boeremia exigua. Their structures with absolute configurations were established by extensive spectroscopic methods and electronic circular dichroism (ECD) calculations. Compounds 1 – 4 , 6 , and 9 displayed anti-inflammatory properties on nitric oxide production in LPS-induced RAW264.7 macrophages with an IC 50 range of 19.4–34.4 μM. Compounds 2 and 5 exhibited cytotoxicities to human breast cancer cell line (MCF–7) with IC 50 values of 33.1 and 4.0 μM, respectively. Unlabelled Image • Three new depsidones and two new diaryl ethers have been isolated from fungus Boeremia exigua. • The new structures with absolute configurations were elucidated by means of spectroscopic methods and ECD calculations. • All compounds were evaluated for their anti-pathogenic activities, NO production inhibitions, and cytotoxicities. [ABSTRACT FROM AUTHOR]

Three newly isolated ergosterols, psathergosterols A–C (1 – 3), together with two known ones (4 and 5), have been isolated from cultures of the basdiomycete Psathyrella candolleana. Their structures with the absolute configuration were elucidated by means of spectroscopic methods and the single crystal X-ray diffraction. Compounds 2 – 4 exhibited certain cytotoxicities to five human cancer cell lines (HL-60, SMMC-7721, A-549, MCF-7, SW480). Unlabelled Image [ABSTRACT FROM AUTHOR]

The title triterpenoid (I) exhibits cytotoxic activity against a number of human cancer cell lines (HL-60, SMMC-7721, A-549, MCF-7, and SW480). [ABSTRACT FROM AUTHOR]

Spermatogonial stem cells (SSCs) have significant applications in both reproductive and regenerative medicine. However, primary human SSCs are very rare, and a human SSC line has not yet been available. In this study, we have for the first time reported a stable human SSC line by stably expressing human SV40 large T antigen. RT-PCR, immunocytochemistry, and Western blots revealed that this cell line was positive for a number of human spermatogonial and SSC hallmarks, including VASA, DAZL, MAGEA4, GFRA1, RET, UCHL1, GPR125, PLZF and THY1, suggesting that these cells are human SSCs phenotypically. Proliferation analysis showed that the cell line could be expanded with significant increases of cells for 1.5 years, and high levels of PCNA, UCHL1 and SV40 were maintained for long-term culture. Transplantation assay indicated that human SSC line was able to colonize and proliferate in vivo in the recipient mice. Neither Y chromosome microdeletions of numerous genes nor tumor formation was observed in human SSC line although there was abnormal karyotype in this cell line. Collectively, we have established a human SSC line with unlimited proliferation potentials and no tumorgenesis, which could provide an abundant source of human SSCs for their mechanistic studies and translational medicine. [ABSTRACT FROM AUTHOR]