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Introduction: Malignant melanoma (MM) is a highly aggressive skin tumour. Aim: To investigate whether miR-22 is involved in the proliferation, invasion, and migration of melanoma cells (MCs) by negatively regulating NOD-like receptor protein 3 (NLRP3) gene. Material and methods: Human MCs (WM239a) and human epidermal melanocytes (HEM) were used as study material. The expression levels of miR-22 and NLRP3 were detected by qRT-PCR. The expression of NLRP3 protein was determined by Western blot (WB) analysis. The effects of miR-22 and NLRP3 on the proliferation, invasion, and migration of MCs were evaluated by cell counting kit-8 (CCK-8), Transwell cell invasion assay, and scratch assay. Results: The expression of miR-22 was clearly lower in WM239a than in HEM. Up-regulation of miR-22 expression in WM239a clearly raised the expression of miR-22, Caspase-1, and E-cadherin and the apoptotic rate of WM239a; however, the levels of interleukin-1β (IL-1β) and NLRP3, cell proliferation activity, invasion and migration ability were clearly decreased. The negative regulation of NLRP3 by miR-22 may play a major role in activities of MM. Conclusions: Further studies will help to reveal the molecular details of this regulatory mechanism and provide new therapeutic strategies. [ABSTRACT FROM AUTHOR]

【Objective】This work aims to investigate the effects and mechanisms of crude extract of F. suspensa leaves tea and its main components, phillyrin and Forsythia glycoside A, on the proliferation and migration function of hepatocellular carcinoma LM3.【Method】 Water extraction method was used to extract green tea and black tea from Forsythia suspensa leaves tea, HPLC was used to detect the content of effective components such as phillyrin and Forsythia glycoside A in F. suspensa green tea and F. suspensa black tea. CCK8 experiment was used to explore the effects of crude extracts of two types of F. suspensa leaves tea, phillyrin and Forsythia glycoside A on the proliferation of hepatocellular carcinoma LM3. Two crude extracts of Forsythia suspensa leaves tea, phillyrin and Forsythia glycoside A, were used to culture hepatocellular carcinoma LM3. The migration of cells was observed at 24, 48, 72, and 96 h, and the effects of the two crude extracts of F. suspensa leaves tea, phillyrin and Forsythia glycoside A on the migration of hepatocellular carcinoma LM3 were detected through the cell scratch test. To reveal the molecular mechanism of crude extract of F. suspensa leaves tea and its main components phillyrin and Forsythia glycoside A affect the proliferation and migration of hepatocellular carcinoma. The expressions of proliferation and migration-related gene Ki-67, Erk, PI3K and mTOR were detected using RT-PCR technology.【Result】Both the green tea and black tea extracts of F. suspensa leaves tea, as well as phillyrin and Forsythia glycoside A, significantly inhibited the proliferation and migration of hepatocellular carcinoma LM3, while Forsythia glycoside A significantly inhibited the migration of hepatocellular carcinoma line LM3. Among them, the crude extract of F. suspensa green tea significantly inhibited the proliferation and migration of HCC at low, medium, and high concentrations when 24, 48, and 72 h. The inhibitory effect of the crude extract of F. suspensa black tea on the proliferation and migration of LM3 cells continued to increase with increasing concentration, and the best inhibitory effect was achieved at high concentration for 72 h. The analysis of RT-PCR results suggests that its main mechanism of action may be achieved by reducing the expression levels of proliferation and migration related genes such as Ki-67.【Conclusion】 In summary, the crude extracts of F. suspensa green tea and F. suspensa black tea, as well as phillyrin and Forsythia glycoside A, can inhibit the proliferation or migration of HCC LM3 by reducing the expression of Ki-67. [ABSTRACT FROM AUTHOR]

Podostroma cornu-damae, one of the lethal toxic mushrooms, is known to contain macrocyclic trichothecene mycotoxins exhibiting potent cytotoxic effects, attracting attention as an important research subject for scientists interested in natural product chemistry and toxicity research. To investigate the mycotoxins from the toxic mushroom P. cornu-damae and evaluate their cytotoxic activities, the fungus was large-cultured on solid plates and successively extracted to acquire a crude methanol (MeOH) extract. After performing successive separation and purification processes, a total of eight macrocyclic trichothecenes were isolated from the MeOH extract of plate cultures of P. cornu-damae using the liquid chromatography/mass spectrometry (LC/MS)-guided isolation technique. Extensive interpretation of nuclear magnetic resonance (NMR) spectroscopic and high-resolution (HR)-electrospray ionization (ESI)-MS data allowed for the structural identification of all isolated macrocyclic trichothecenes, including satratoxin I (1), satratoxin H (2), roridin E (3), miophytocen D (4), roridin L-2 (5), trichoverritone (6), 12′-episatratoxin H (7), and roridin F (8). We conducted a cytotoxicity evaluation of compounds 1–8 against 4T1 breast cancer cells and fibroblast cell lines (L929 cells) using the Counting Kit-8 (CCK-8) cell viability assay to validate their cytotoxic potential. Our results indicated that compounds 1–6 lack anti-cancer effects on 4T1 cells and have minimal impact on the viability of the fibroblast cell line, L929 cells. In contrast, compounds 7 and 8 exhibited no cytotoxicity in normal cells (L929) and demonstrated specific cytotoxicity in breast cancer cell lines. Notably, the cytotoxic effects of compounds 7 and 8 in 4T1 cells were significantly stronger than those observed with free doxorubicin. These findings suggest that compounds 7 and 8 may possess targeted anti-cancer effects, specifically against breast cancer cells, emphasizing their efficient and selective toxicity towards breast cancer cells. [ABSTRACT FROM AUTHOR]

In the current study, a series of 1,3,4-oxadiazole and 1,2,4-triazole derivatives linked to quinazolinone were designed and synthesized by using Phase Transfer Catalysis. The structures of the synthesized compounds were confirmed by their IR, 1H NMR, 13C NMR, and HR-ESI-MS spectroscopic data, and their in vitro antiproliferative activities against A549, Hela, MCF-7 as well as L929 were detected by using CCK-8 assay. The results demonstrated that most of the compounds showed better growth inhibitory effect than 5-FU on A549, Hela, and MCF-7 cell lines. Especially, 2-(5-((3-bromophenyl)thio)-1,3,4-oxadiazole-2-yl)quinazolin-4(3H)-one (8o), 4-amino-5-((4-nitrophenyl)thio)-4H-1,2,4-triazol-3-yl)quinazolin-4(3H)-one (9m) and 2-(4-amino-5-(o-tolylthio)-4H-1,2,4-triazol-3-yl)quinazolin-4(3H)-one (9b) showed good antiproliferative activity against A549 cells with IC50 value of 3.46, 2.96, and 3.44 μM, respectively. 2-(4-Amino-5-((3,5-difluorophenyl)thio)-4H-1,2,4-triazol-3-yl)quinazolin-4(3H)-one (9h) and 4-amino-5-((4-nitrophenyl)thio)-4H-1,2,4-triazol-3-yl)quinazolin-4(3H)-one (9m) had obvious inhibitory effects on Hela cells with IC50 values of 1.76 and 3.91 μM, respectively. 2-(4-Amino-5-((2-chlorophenyl)thio)-4H-1,2,4-triazol-3-yl)quinazolin-4(3H)-one (9i) showed high inhibitory activity against MCF-7 cells with IC50 value of 2.72 μM. Meanwhile, all compounds showed lower cytotoxicity to L929 cells than positive control drug 5-FU. In development, according to the above preliminary observation, these novel quinazolinones incorporating 1,3,4-oxadiazole and 1,2,4-triazole thioether moieties could become potential molecular templates for searching new antitumor agents. [ABSTRACT FROM AUTHOR]

Trichomonas gallinae is a protozoan parasite that is the causative agent of trichomoniasis, and infects captive and wild bird species throughout the world. Although metronidazole has been the drug of choice against trichomoniasis for decades, most Trichomonas gallinae strains have developed resistance. Therefore, drugs with new modes of action or targets are urgently needed. Here, we report the development and application of a cell-based CCK-8 method for the high-throughput screening and identification of new inhibitors of Trichomonas gallinae as a beginning point for the development of new treatments for trichomoniasis. We performed the high-throughput screening of 173 anti-parasitic compounds, and found 16 compounds that were potentially effective against Trichomonas gallinae. By measuring the median inhibitory concentration (IC50) and median cytotoxic concentration (CC50), we identified 3 potentially safe and effective compounds against Trichomonas gallinae: anisomycin, fumagillin, and MG132. In conclusion, this research successfully established a high-throughput screening method for compounds and identified 3 new safe and effective compounds against Trichomonas gallinae, providing a new treatment scheme for trichomoniasis.

Trichomonas gallinae is a protozoan parasite that is the causative agent of trichomoniasis, and infects captive and wild bird species throughout the world. Although metronidazole has been the drug of choice against trichomoniasis for decades, most Trichomonas gallinae strains have developed resistance. Therefore, drugs with new modes of action or targets are urgently needed. Here, we report the development and application of a cell-based CCK-8 method for the high-throughput screening and identification of new inhibitors of Trichomonas gallinae as a beginning point for the development of new treatments for trichomoniasis. We performed the high-throughput screening of 173 anti-parasitic compounds, and found 16 compounds that were potentially effective against Trichomonas gallinae. By measuring the median inhibitory concentration (IC 50) and median cytotoxic concentration (CC 50), we identified 3 potentially safe and effective compounds against Trichomonas gallinae : anisomycin, fumagillin, and MG132. In conclusion, this research successfully established a high-throughput screening method for compounds and identified 3 new safe and effective compounds against Trichomonas gallinae , providing a new treatment scheme for trichomoniasis. [Display omitted] • A high-throughput screening method was developed for compounds against T. gallinae. • High-throughput screening method based on CCK-8 was superior to resazurin method. • Fumagillin, anisomycin and MG132 are potential candidate drugs against T. gallinae. [ABSTRACT FROM AUTHOR]

Molecularly imprinted polymers and non‐imprinted polymers based on attapulgite and graphene composite were synthesized by coating with copolymer of acrylamide and methyl methacrylate. The adsorption results showed that the molecularly imprinted polymers and non‐imprinted polymers could reach the maximum drug loading of 2.4 and 1.6 mg/g, respectively. The in vitro release studies demonstrated that molecularly imprinted polymers and non‐imprinted polymers can control the release of 5‐fluorouracil and reach maximum release at approximately 48 h. Cytotoxicity analysis showed that 100 μg/mL of molecularly imprinted polymers loaded with 5‐fluorouracil gradually increased the inhibition rate of HeLa with increasing time and approached the maximum inhibition rate of 20 % at 72 h. [ABSTRACT FROM AUTHOR]

We prepared a visual Rhodamine B-nido-carborane fluorescent marker by one-pot method for tumor cell imaging. Firstly, the fat-soluble carborane was treated with potassium hydroxide to obtain nido-carborane potassium salt. Secondly, it was combined with Rhodamine B to remove a molecule of potassium chloride, and finally a fluorescent marker of Rhodamine B-nido-carborane was obtained. In order to further understand its structural characteristics, it was observed that the nido-carborane and Rhodamine B are firmly bound together by ionic properties by X-ray single crystal diffraction. Such a design strategy is conducive to combining the advantages of the two, and can better promote its application in the biological field through the synergistic action of complementarity and mutual assistance. The addition rate and inhibition rate of Rhodamine B-nido-carborane in Hera cells were measured by CCK-8, and their maximum values were 71.76% and 28.24%, respectively. In fluorescence imaging, HeLa cells were selected to test their biocompatibility in different channels, and it was found that Rhodamine B-nido-carborane had a good labeling effect on tumor cells. [ABSTRACT FROM AUTHOR]

A growing body of evidence suggests that anesthetics impact the outcome of patients with cancer after surgical intervention. However, the optimal dose and underlying mechanisms of co-administered anesthetics in lung tumor therapy have been poorly studied. Here, we aimed to investigate the role of combined anesthetics propofol, sufentanil, and rocuronium in treating lung cancer using an orthogonal experimental design and to explore the optimal combination of anesthetics. First, we evaluated the effects of the three anesthetics on the proliferation and invasion of A-549 cells using Cell Counting Kit 8 and Transwell migration and invasion assays. Subsequently, we applied the orthogonal experimental design (OED) method to screen the appropriate concentrations of the combined anesthetics with the most effective antitumor activity. We found that all three agents inhibited the proliferation of A-549 cells in a dose- and time-dependent manner when applied individually or in combination, with the highest differences in the magnitude of inhibition occurring 24 h after combined drug exposure. The optimal combination of the three anesthetics that achieved the strongest reduction in cell viability was 1.4 mmol/L propofol, 2 nmol/L sufentanil, and 7.83 mmol/L rocuronium. This optimal 3-drug combination produced a more beneficial result at 24 h than either single drug. Our results provide a theoretical basis for improving the efficacy of lung tumor treatment and optimizing anesthetic strategies. [ABSTRACT FROM AUTHOR]

Two new Ag(I) coordination polymers, namely [Ag(bpp)]·0.5 n(1,5-NDSA)·n(H2O) (1) and [Ag2(bpp)2]n·n(2,7-NDSA)·2 n(H2O)·n(CH3CN) (2) (Na2(1,5-NDSA) = sodium 1,5-naphthalenedisulfonate dibasic, Na2(2,7-NDSA) = sodium 1,5-naphthalenedisulfonate dibasic, bpp is 1,3-bis(4-pyridyl)propane), were generated via the solution evaporation method under room temperature. Moreover, the solids of these two compounds display strong luminescence emission at RT. And the application values of the compounds against the glioblastoma treatment were determined, and the corresponding mechanism was simultaneously tested. The analysis of CCK-8 was first implemented and the glioblastoma viability was measured. The real-time RT-PCR was next performed, and the signaling pathway activation of VEGF in glioblastoma cells was tested after treating by the above compound.

Introduction: The actin regulatory protein, cofilin plays a key signaling role in many cells for numerous cellular responses including in proliferation, development, motility, migration, secretion and growth. In the pancreas it is important in islet insulin secretion, growth of pancreatic cancer cells and in pancreatitis. However, there are no studies on its role or activation in pancreatic acinar cells. Methods: To address this question, we studied the ability of CCK to activate cofilin in pancreatic acinar cells, AR42J cells and CCK1-R transfected Panc-1 cells, the signaling cascades involved and its effect on enzyme secretion and MAPK activation, a key mediator of pancreatic growth. Results: CCK (0.3 and 100 nM), TPA, carbachol, Bombesin, secretin and VIP decreased phospho-cofilin (i.e., activate cofilin) and both phospho-kinetic and inhibitor studies of cofilin, LIM kinase (LIMK) and Slingshot Protein Phosphatase (SSH1) demonstrated these conventional activators of cofilin were not involved. Serine phosphatases inhibitors (calyculin A and okadaic acid), however inhibited CCK/TPA-cofilin activation. Studies of various CCK-activated signaling cascades showed activation of PKC/PKD, Src, PAK4, JNK, ROCK mediated cofilin activation, but not PI3K, p38, or MEK. Furthermore, using both siRNA and cofilin inhibitors, cofilin activation was shown to be essential for CCK-mediated enzyme secretion and MAPK activation. Conclusion: These results support the conclusion that cofilin activation plays a pivotal convergent role for various cell signaling cascades in CCK mediated growth/enzyme secretion in pancreatic acini. [ABSTRACT FROM AUTHOR]

Podostroma cornu-damae, one of the lethal toxic mushrooms, is known to contain macrocyclic trichothecene mycotoxins exhibiting potent cytotoxic effects, attracting attention as an important research subject for scientists interested in natural product chemistry and toxicity research. To investigate the mycotoxins from the toxic mushroom P. cornu-damae and evaluate their cytotoxic activities, the fungus was large-cultured on solid plates and successively extracted to acquire a crude methanol (MeOH) extract. After performing successive separation and purification processes, a total of eight macrocyclic trichothecenes were isolated from the MeOH extract of plate cultures of P. cornu-damae using the liquid chromatography/mass spectrometry (LC/MS)-guided isolation technique. Extensive interpretation of nuclear magnetic resonance (NMR) spectroscopic and high-resolution (HR)-electrospray ionization (ESI)-MS data allowed for the structural identification of all isolated macrocyclic trichothecenes, including satratoxin I (1), satratoxin H (2), roridin E (3), miophytocen D (4), roridin L-2 (5), trichoverritone (6), 12′-episatratoxin H (7), and roridin F (8). We conducted a cytotoxicity evaluation of compounds 1–8 against 4T1 breast cancer cells and fibroblast cell lines (L929 cells) using the Counting Kit-8 (CCK-8) cell viability assay to validate their cytotoxic potential. Our results indicated that compounds 1–6 lack anti-cancer effects on 4T1 cells and have minimal impact on the viability of the fibroblast cell line, L929 cells. In contrast, compounds 7 and 8 exhibited no cytotoxicity in normal cells (L929) and demonstrated specific cytotoxicity in breast cancer cell lines. Notably, the cytotoxic effects of compounds 7 and 8 in 4T1 cells were significantly stronger than those observed with free doxorubicin. These findings suggest that compounds 7 and 8 may possess targeted anti-cancer effects, specifically against breast cancer cells, emphasizing their efficient and selective toxicity towards breast cancer cells.

Abstract Ovarian cancer is one of the most common gynecological tumors that seriously endanger the health and quality of life of women. Microfibril‐associated protein 2 (MFAP2) has been demonstrated to play crucial roles in the development of multiple tumors. However, the function of MFAP2 in ovarian cancer remains unclear. In this study, we found that MFAP2 was upregulated in ovarian cancer and cells and was positively correlated with FOXM1 and glycolysis‐related genes. The results of Cell Count Kit‐8, colony formation, and flow cytometry assays indicated that MFAP2 promoted cell proliferation. In addition, MFAP2 promotes cell proliferation, glucose uptake, lactate production; increases ATP levels, extracellular acidification ratio, and oxygen consumption ratio in ovarian cancer cells and increases the expression of glycolytic proteins. Further mechanistic analysis suggests that MFAP2 promotes FOXM1/β‐catenin‐mediated glycolysis signaling in ovarian cancer cells. Knockdown of MFAP2 inhibits ovarian cancer xenograft tumor growth and expression of Ki‐67, MFAP2, FOXM1, GLUT1, HK2, and β‐catenin in mice. In conclusion, MFAP2 promotes cell proliferation and glycolysis by modulating the FOXM1/β‐catenin signaling pathway in ovarian cancer, which may offer a fresh insight into the treatment of ovarian cancer in the glycolysis pathway.

Introduction: The actin regulatory protein, cofilin plays a key signaling role in many cells for numerous cellular responses including in proliferation, development, motility, migration, secretion and growth. In the pancreas it is important in islet insulin secretion, growth of pancreatic cancer cells and in pancreatitis. However, there are no studies on its role or activation in pancreatic acinar cells.Methods: To address this question, we studied the ability of CCK to activate cofilin in pancreatic acinar cells, AR42J cells and CCK1-R transfected Panc-1 cells, the signaling cascades involved and its effect on enzyme secretion and MAPK activation, a key mediator of pancreatic growth.Results: CCK (0.3 and 100 nM), TPA, carbachol, Bombesin, secretin and VIP decreased phospho-cofilin (i.e., activate cofilin) and both phospho‐kinetic and inhibitor studies of cofilin, LIM kinase (LIMK) and Slingshot Protein Phosphatase (SSH1) demonstrated these conventional activators of cofilin were not involved. Serine phosphatases inhibitors (calyculin A and okadaic acid), however inhibited CCK/TPA-cofilin activation. Studies of various CCK‐activated signaling cascades showed activation of PKC/PKD, Src, PAK4, JNK, ROCK mediated cofilin activation, but not PI3K, p38, or MEK. Furthermore, using both siRNA and cofilin inhibitors, cofilin activation was shown to be essential for CCK-mediated enzyme secretion and MAPK activation.Conclusion: These results support the conclusion that cofilin activation plays a pivotal convergent role for various cell signaling cascades in CCK mediated growth/enzyme secretion in pancreatic acini.

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AIM: To determine the role of heparanase-1 (HPSE-1) in orbital rhabdomyosarcoma (RMS), and to investigate the feasibility of HPSE-1 targeted therapy for RMS. METHODS: Immunohistochemistry was performed to analyze HPSE-1 expression in 51 cases of orbital RMS patients (including 28 cases of embryonal RMS and 23 cases of alveolar RMS), among whom there were 27 treated and 24 untreated with preoperative chemoradiotherapy. In vitro, studies were conducted to examine the effect of HPSE-1 silencing on RMS cell proliferation and tube formation of human umbilical vein endothelial cells (HUVECs). RD cells (an RMS cell line) and HUVECs were infected with HPSE-1 shRNA lentivirus at a multiplicity of infection (MOI) of 10 and 30 separately. Real-time PCR and Western blot were applied to detect the mRNA and protein expression levels of HPSE-1. Cell viability of treated or control RD cells was evaluated by cell counting kit-8 (CCK-8) assay. Matrigel tube formation assay was used to evaluate the effect of HPSE-1 RNAi on the tube formation of HUVECs. RESULTS: Immunohistochemistry showed that the expression rate of HPSE-1 protein was 92.9% in orbital embryonal RMS and 91.3% in orbital alveolar RMS. Tissue from alveolar orbital RMS did not show relatively stronger staining than that from the embryonal orbital RMS. However, despite the types of RMS, comparing the cases treated chemoradiotherapy with those untreated, we have observed that chemoradiotherapy resulted in weaker staining in patients' tissues. The expression levels of HPSE-1 declined significantly in both the mRNA and protein levels in HPSE-1 shRNA transfected RD cells. The CCK-8 assay showed that lentivirus-mediated HPSE-1 silencing resulted in significantly reduced RD cells viability in vitro. Silencing HPSE-1 expression also inhibited VEGF-induced tube formation of HUVECs in Matrigel. CONCLUSION: HPSE-1 silencing may be a promising therapy for the inhibition of orbital RMS progression.

Two new Ag(I) coordination polymers, namely [Ag(bpp)]·0.5 n(1,5-NDSA)·n(H2O) (1) and [Ag2(bpp)2]n·n(2,7-NDSA)·2 n(H2O)·n(CH3CN) (2) (Na2(1,5-NDSA) = sodium 1,5-naphthalenedisulfonate dibasic, Na2(2,7-NDSA) = sodium 1,5-naphthalenedisulfonate dibasic, bpp is 1,3-bis(4-pyridyl)propane), were generated via the solution evaporation method under room temperature. Moreover, the solids of these two compounds display strong luminescence emission at RT. And the application values of the compounds against the glioblastoma treatment were determined, and the corresponding mechanism was simultaneously tested. The analysis of CCK-8 was first implemented and the glioblastoma viability was measured. The real-time RT-PCR was next performed, and the signaling pathway activation of VEGF in glioblastoma cells was tested after treating by the above compound. [ABSTRACT FROM AUTHOR]

In the current study, a new Cu(II)-containing coordination polymer (CP) with the chemical formula of {[Cu2(H2O)5(tpt)(L)]·6H2O}n (1, tpt = 2,4,6‐tris(4‐pyridyl)‐1,3,5‐triazine and L = 3,3′‐disulfonyl‐4,4′‐biphenyldicarboxylate) has been successfully produced via a hydrothermal reaction of Cu(NO3)2·3H2O with the carboxylate ligand L in the presence of the auxiliary N-donor ligand tpt. The anti-cancer activity of the new compound on retinoblastoma was evaluated and the related mechanism was explored at the same time. First of all, the Cell Counting Kit-8 (CCK-8) kit was performed to determine the viability of the Y79 retinoblastoma cells after compound treatment. Next, after compound exposure, the activation of the JAK-STA signaling pathway in the Y79 retinoblastoma cells was measured with real time reverse transcription-polymerase chain reaction (RT-PCR) assay. [ABSTRACT FROM AUTHOR]

Ovarian cancer is one of the most common gynecological tumors that seriously endanger the health and quality of life of women. Microfibril‐associated protein 2 (MFAP2) has been demonstrated to play crucial roles in the development of multiple tumors. However, the function of MFAP2 in ovarian cancer remains unclear. In this study, we found that MFAP2 was upregulated in ovarian cancer and cells and was positively correlated with FOXM1 and glycolysis‐related genes. The results of Cell Count Kit‐8, colony formation, and flow cytometry assays indicated that MFAP2 promoted cell proliferation. In addition, MFAP2 promotes cell proliferation, glucose uptake, lactate production; increases ATP levels, extracellular acidification ratio, and oxygen consumption ratio in ovarian cancer cells and increases the expression of glycolytic proteins. Further mechanistic analysis suggests that MFAP2 promotes FOXM1/β‐catenin‐mediated glycolysis signaling in ovarian cancer cells. Knockdown of MFAP2 inhibits ovarian cancer xenograft tumor growth and expression of Ki‐67, MFAP2, FOXM1, GLUT1, HK2, and β‐catenin in mice. In conclusion, MFAP2 promotes cell proliferation and glycolysis by modulating the FOXM1/β‐catenin signaling pathway in ovarian cancer, which may offer a fresh insight into the treatment of ovarian cancer in the glycolysis pathway. [ABSTRACT FROM AUTHOR]

Combined activation of GLP-1 and CCK1 receptors has potential to synergistically augment the appetite-suppressive and glucose homeostatic actions of the individual parent peptides. In the current study, pancreatic beta-cell benefits of combined GLP-1 and CCK1 receptor upregulation were established, before characterising bioactivity and antidiabetic efficacy of an acylated dual-acting GLP-1/CCK hybrid peptide, namely [Lys12Pal]Ex-4/CCK. Both exendin-4 and CCK exhibited (p<0.001) proliferative and anti-apoptotic effects in BRIN BD11 beta-cells. Proliferative benefits were significantly (p<0.01) augmented by combined peptide treatment when compared to either parent peptide alone. These effects were linked to increases (p<0.001) in GLUT2 and glucokinase beta-cell gene expression, with decreased (p<0.05-p<0.001) expression of NFκB and BAX. [Lys12Pal]Ex-4/CCK exhibited prominent insulinotropic actions in vitro, coupled with beneficial (p<0.001) satiety and glucose homeostatic effects in the mice, with bioactivity evident 24 h after administration. Following twice daily injection of [Lys12Pal]Ex-4/CCK for 28 days in diabetic high fat fed (HFF) mice with streptozotocin (STZ)-induced compromised beta-cells, there were clear reductions (p<0.05-p<0.001) in energy intake and body weight. Circulating glucose was returned to lean control concentrations, with associated increases (p<0.001) in plasma and pancreatic insulin levels. Glucose tolerance and insulin secretory responsiveness were significantly (p<0.05-p<0.001) improved by hybrid peptide therapy. In keeping with this, evaluation of pancreatic histology revealed restoration of normal islet alpha- to beta-cell ratios and reduction (p<0.01) in centralised islet glucagon staining. Improvements in pancreatic islet morphology were associated with increased (p<0.05) proliferation and reduced (p<0.001) apoptosis of beta-cells. Together, these data highlight the effectiveness of sustained dual GLP-1 and CCK1 receptor activation by [Lys12Pal]Ex-4/CCK for the treatment of obesity-related diabetes. [ABSTRACT FROM AUTHOR]

By using a mixed-ligand approach, a new Cu(II)-based coordination polymer (CP) with the chemical formula of {[Cu2(ptd)2(IPA)2]·2H2O}n (1, ptd = 4′-(4-pyridine)4,2′:2′,4′′-terpyridine, H2IPA = isophthalic acid) has been synthesized under hydrothermal conditions and characterized by single-crystal X-ray diffraction, IR spectroscopic, elemental and thermogravimetric analyses. Furthermore, the anticancer activity of the compound on the CHO cancer cell was determined, the CCK-8 assay was performed to explore the inhibitory activity of the compound on the cell viability. the IC50 values were calculated at the same time. The suppression of the compound on the cancer cell migration and invasion ability was also determined with trans-well assay. [ABSTRACT FROM AUTHOR]

Individual differences in the effects of a chronic neuropathic injury on social behaviours characterize both the human experience and pre‐clinical animal models. The impacts of these changes to the well‐being of the individual are often underappreciated. Earlier work from our laboratory using GeneChip® microarrays identified increased cholecystokinin (CCK) gene expression in the periaqueductal gray (PAG) of rats that showed persistent changes in social interactions during a Resident–Intruder encounter following sciatic nerve chronic constriction injury (CCI). In this study, we confirmed these gene regulation patterns using RT‐PCR and identified the anatomical location of the CCK‐mRNA as well as the translated CCK peptides in the midbrains of rats with a CCI. We found that rats with persistent CCI‐induced changes in social behaviours had increased CCK‐mRNA in neurons of the ventrolateral PAG and dorsal raphe nuclei, as well as increased CCK‐8 peptide expression in terminal boutons located in the lateral and ventrolateral PAG. The functional significance of these changes was explored by microinjecting small volumes of CCK‐8 into the PAG of uninjured rats and observing their Resident–Intruder social interactions. Disturbances to social interactions identical to those observed in CCI rats were evoked when injection sites were located in the rostral lateral and ventrolateral PAG. We suggest that CCI‐induced changes in CCK expression in these PAG regions contributes to the disruptions to social behaviours experienced by a subset of individuals with neuropathic injury. [ABSTRACT FROM AUTHOR]

We studied the effect of CCK-8 on BP and blood content of CGRP in rats with hypertension caused by fructose or inhibition of NO synthase with L-NAME. The decrease in the CGRP content was found during the development of fructose-induced hypertension, but not L-NAME-caused hypertension. Administration of CCK-8 to fructose-fed animals reduced BP and increased the content of CGRP. In rats with hypertension caused by NO deficit, CCK-8 lowered BP, but did not affect the content of CGRP. These findings suggest that CGRP mediates the hypotensive effect of CCK-8 in fructose-induced hypertension, but not in NO-deficient hypertension. [ABSTRACT FROM AUTHOR]

Aristolochic acid (AA) is widely present in herbal medicine. Aristolochic acid I (AAI) and aristolochic acid lactam (AAT), as the main active ingredients in AAs, have serious nephrotoxicity, strong carcinogenicity, and mutagenicity. AAT is a metabolite of AAI in the human body. AAI can be reduced by Fe2+ both in the human body and soil. A new dual spectrum is proposed to monitor the reaction directly from the human protein serum (HAS) and A549 cells through surface‐enhanced Raman spectroscopy (SERS) and its biological product AAT through fluorescence detection (FLD) spectra based on its unique spectral response. In order to better monitor the reduction reaction, the AAI detection conditions are optimized, including concentration, pH, reducing agent, and reaction time. During the experiment, Ag@Au is selected as the SERS substrate. Moreover, scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were used to characterize the prepared SERS support substrate. CCK‐8 assays showed that the viability of cells decline after adding AAI. Fe2+ was found to detect AAI in endogenous A 549 cells. [ABSTRACT FROM AUTHOR]

Through applying the mixed-ligand method, a new coordination polymer (CP) of Cu(II) with the chemical formula of {Cu5(L)2(1,4-bimb)(μ3-OH)2(H2O)8]·2H2O}n (1, 1,4-bimb = 1,4-bis(imidazol-1-ylmethyl)benzene and H4L = 1,2,4,5-cyclohexanetetracarboxylic acid) was synthesized under the solvothermal conditions. Furthermore, the biological effect of compound against glomerulonephritis therapy was detected, and the details mechanism was investigated. First of all, the mesangial cells proliferation was detected through using the Cell Counting Kit-8 (CCK-8) method. Then, the Annexin V-FITC/PI method was utilized to determine cell cycle arrest and the apoptosis rate. Next, the caspase-8 and caspase-3 expression level in mesangial cells was detected via applying the western blot method. Finally, the inflammatory cytokines released into the plasma was assessed by ELISA detection kit. [ABSTRACT FROM AUTHOR]

Combined activation of GLP-1 and CCK1 receptors has potential to synergistically augment the appetite-suppressive and glucose homeostatic actions of the individual parent peptides. In the current study, pancreatic beta-cell benefits of combined GLP-1 and CCK1 receptor upregulation were established, before characterising bioactivity and antidiabetic efficacy of an acylated dual-acting GLP-1/CCK hybrid peptide, namely [Lys12Pal]Ex-4/CCK. Both exendin-4 and CCK exhibited (p

Combined activation of GLP-1 and CCK1 receptors has potential to synergistically augment the appetite-suppressive and glucose homeostatic actions of the individual parent peptides. In the current study, pancreatic beta-cell benefits of combined GLP-1 and CCK1 receptor upregulation were established, before characterising bioactivity and antidiabetic efficacy of an acylated dual-acting GLP-1/CCK hybrid peptide, namely [Lys12Pal]Ex-4/CCK. Both exendin-4 and CCK exhibited (p<0.001) proliferative and anti-apoptotic effects in BRIN BD11 beta-cells. Proliferative benefits were significantly (p<0.01) augmented by combined peptide treatment when compared to either parent peptide alone. These effects were linked to increases (p<0.001) in GLUT2 and glucokinase beta-cell gene expression, with decreased (p<0.05-p<0.001) expression of NFκB and BAX. [Lys12Pal]Ex-4/CCK exhibited prominent insulinotropic actions in vitro , coupled with beneficial (p<0.001) satiety and glucose homeostatic effects in the mice, with bioactivity evident 24 h after administration. Following twice daily injection of [Lys12Pal]Ex-4/CCK for 28 days in diabetic high fat fed (HFF) mice with streptozotocin (STZ)-induced compromised beta-cells, there were clear reductions (p<0.05-p<0.001) in energy intake and body weight. Circulating glucose was returned to lean control concentrations, with associated increases (p<0.001) in plasma and pancreatic insulin levels. Glucose tolerance and insulin secretory responsiveness were significantly (p<0.05-p<0.001) improved by hybrid peptide therapy. In keeping with this, evaluation of pancreatic histology revealed restoration of normal islet alpha- to beta-cell ratios and reduction (p<0.01) in centralised islet glucagon staining. Improvements in pancreatic islet morphology were associated with increased (p<0.05) proliferation and reduced (p<0.001) apoptosis of beta-cells. Together, these data highlight the effectiveness of sustained dual GLP-1 and CCK1 receptor activation by [Lys12Pal]Ex-4/CCK for the treatment of obesity-related diabetes. [ABSTRACT FROM AUTHOR]

To compare the use of real-time cell analysis system (RTCA) and Cell Counting Kit-8 (CCK-8) in detecting the toxic effects of diesel exhaust particles (DEP) on human bronchial epithelial (HBE) cells, the HBE cells were exposed to two types of Standard Reference Materials (SRM 1650b and SRM 2975) with the exposure concentrations of (0, 3.5, 7, 14, 28 and 56 mg⋅L-1). After exposure for (6, 12, 24 and 48 h), the cytotoxicity of HBE cells induced by different DEP was detected. Then the advantages or shortcomings of RTCA and CCK-8 were analyzed by comparing the cell viability and normalized cell index (NCI) of different groups. And the cell apoptosis assay was used to detect the apoptosis rate between the different groups. Results show that: SRM 2975 was more toxic to HBE cells than SRM 1650b at the same exposure concentration and time. When RTCA was used to detect DEP-induced HBE cytotoxicity, the NCI value of low concentration DEP groups showed statistically significant difference (P <0.05). In comparison, the significant toxicity effects (P<0.05) were detected in relatively higher concentrations of DEP at the same exposure time by use of CCK-8 assay. And the cell apoptosis assay confirmed that the apoptosis rate of low concentration of DEP groups had already showed statistical difference compared with the control group (P< 0.05). It is indicated that compared with CCK-8, RTCA was more suitable for detecting cytotoxicity of DEP-induced adherent HBE cells. And CCK-8 assays could be a better choice for detecting DEP-induced cytotoxicity of suspended cells or adhesion/suspension cytotoxicity when combined with air-liquid interface exposure systems. [ABSTRACT FROM AUTHOR]

By using a mixed-ligand approach, two new Cu(II)-containing co-ordination polymers (CPs) with the chemical formula of [Cu4(HDNA)4(4,4′-bpy)(OH)2]·4H2O·2CH3CN (1, H2DNA = 4,4′-dicarboxy-4″-nitrotriphenyl-amine, 4,4′-bpy = 4,4′-bipyridine) and [Cu3(TCA)2(DPE)]·10DMF·6H2O (2, H3TCA = 4,4′,4″-nitrilotribenzoic acid, dpe = (E)-1,2-di(pyridin-4-yl)diazene) have been successfully prepared by reaction of Cu(NO3)2·3H2O with the corresponding organic ligands under the solvothermal conditions. The cancer cell viability and proliferation were determined with Cell Counting Kit-8 (CCK-8) assay and the colony formation assay. The migration and invasion capability of A375 melanoma cells under Cu(II)-based co-ordination polymers treatment was evaluated with trans-well assay. [ABSTRACT FROM AUTHOR]

Skeletal muscle plays an important role in the growth and development of meat animals. MicroRNAs (miRNAs) can participate in the regulation of muscle development-related functions; however, there have been few reports on whether there are related miRNAs that conservatively regulate muscle development among different species. In this study, the miRNA transcriptome sequencing data of the muscle tissue of cattle, rat, goat, and pig showed that miR-24-3p may conservatively regulate muscle development in these species. Furthermore, mmu-miR-24-3p can positively regulate C2C12 cell proliferation and apoptosis by regulating key proliferation and apoptosis genes in muscle development, which was verified by CCK-8 and RT-qPCR. Bta-miR-24-3p can also positively regulate the proliferation and apoptosis of bovine muscle primary cells by regulating key proliferation and apoptosis genes in the process of muscle development, as verified by CCK-8 and RT-qPCR. The target genes of miR-24-3p in cattle, rat, goat, and pig, which include a large proportion of target genes shared among the four species, are enriched in multiple cell functions and signal pathways that are closely related to muscle development, as revealed by GO and KEGG enrichment analysis. A double luciferase test showed that the shared target genes WNT4, CAMK2B, and TCF7 were targeted by mmu-miR-24-3p in rat and bta-miR-24-3p in cattle. These three shared target genes WNT4, CAMK2B, and TCF7 are involved in the Wnt signaling pathway, which showed that miR-24-3p plays an important role in rat and cattle. The shared target gene (CAMK2B) in rat and cattle increased significantly after the inhibition of miR-24-3p by RT-qPCR. The findings of this study contribute to a better understanding of the role of miR-24-3p in the regulation of muscle development.

AIM: To investigate the effects of blue light and white light on the proliferation of human retinal pigment epithelial cell line(ARPE-19)at different times, and lay a foundation for further detecting the changes of related factors during photodamage and further studying the signal transduction mechanism during light damage. METHODS:Well-grown ARPE-19 cells were collected for experimentation. The standard curve of CCK-8 was made to determine the proper cell density of ARPE-19 cells and the reaction time of CCK-8 reagent. The cells were divided into dark group, blue light group and white light group, which were irradiated for 3, 6 and 9h respectively. After 12h of light-repellent treatment, CCK-8 method was used to examine the effects of different light sources on the proliferation rate of ARPE-19 cells at different times. RESULTS: The number of cells per well was selected by CCK-8 standard curve to be 20 000, and the corresponding absorbance value was measured after 4h of the action of CCK-8 solution. The CCK-8 test results showed that the cell proliferation rates of the three groups were significantly different(PPPPP=0.253, 0.120). The proliferation rate of cells under white light for 3-6h showed a downward trend, while that of cells under light for 6-9h showed an upward trend. At the same illumination time, the proliferation rate of the cells in the blue and white groups was lower than that in the dark group, and the cell proliferation rate in the blue group was lower than that in the white group. The difference was statistically significant(PCONCLUSION: The proliferation of ARPE-19 cells was inhibited by light irradiation. The proliferation rate of cells in blue light group was significantly lower than that of white light group. With the increase of light time, the cell proliferation rate decreased.

Objective: To verify whether icariin has drug toxicity to osteoblasts and its pre protective effect on inflammatory osteoblasts induced by lipopolysaccharide. Methods: (1) The osteoblasts of newborn SD rats were extracted, cultured and passaged. By observing the morphology of osteoblasts and ALP staining, the activity and proliferation of osteoblasts were determined. (2) CCK-8 method was used to observe the effect of Icariin on osteoblast activity. (3) Different concentrations of lipopolysaccharide (LPS) were used to induce inflammatory osteoblasts in fetal rats. CCK-8 method was used to select the best concentration for induction.(4) This experiment was divided into control group, low concentration group, middle concentration group and high concentration group. CCK-8 method was used to observe whether icariin could protect osteoblasts from inflammatory reaction. Results: (1) The number of osteoblasts in the third generation increased, the shape of osteoblasts overlapped like tiles, most of osteoblasts grew in the center, the center was dense and the specific shape was difficult to see. After ALP staining, the positive cells showed gray black granules in cytoplasm and irregular cell body shape under inverted microscope; (2) DWhen the concentration of icariin was lower than 1 µ g / ml, it had no significant effect on osteoblasts (P < 0.05); when it was higher than 10 µ g / ml, icariin had no significant effect on osteoblasts (P < 0.05); (3) When the concentration of lipopolysaccharide was higher than 80 µ g / ml, there was a significant trend of inflammatory damage to osteoblasts, which was statistically significant (P < 0.01). Therefore, 80 µ g / ml was selected as the best injury concentration in this experiment;(4) When the concentration of icariin was lower than 1 µ g / ml, there was no significant pre protective effect on the inflammatory response of osteoblasts (P < 0.05); when it was higher than 10 µ g / ml, there was significant pre protective effect on the inflammatory response of osteoblasts (P < 0.05). Conclusion: When the concentration of icariin reaches a certain level, it can promote the proliferation of osteoblasts and has a certain pre protective effect on inflammatory osteoblasts. [ABSTRACT FROM AUTHOR]

BACKGROUND: Studies have shown that abnormal osteoblast metabolism may cause abnormal subchondral bone mineralization in knee osteoarthritis. OBJECTIVE: To verify the effects of catalpol from the root of Rehmannia glutinosa on osteoblast proliferation and drug toxicity, and the anti-inflammatory protective effect on inflammatory osteoblasts induced by lipopolysaccharide. METHODS: (1) Osteoblasts from newborn Sprague-Dawley rats were primarily extracted, cultured, and passaged. The activity and proliferation of osteoblasts were determined by observing the morphology of osteoblasts alkaline phosphatase staining and staining of mineralized nodules. (2) Cell counting kit-8 (CCK-8) method was used to observe the effect of different concentrations of catalpol from the root of Rehmannia glutinosa on osteoblast activity. (3) Different concentrations of lipopolysaccharide (0, 10, 20, 40, 80, 160 mg/L) were used to induce inflammatory osteoblasts in fetal rats. CCK-8 method was used to verify the best concentration. (4) The experiment was divided into three groups: blank group, model group, low-, medium- and high-concentration catalpol groups. Inflammatory osteoblasts were induced by lipopolysaccharide at 80 mg/L, and then treated for 8 hours. CCK-8 method was used to observe the effect of different concentrations of catalpol on inflammatory osteoblasts. The study protocol was approved by the Medical Ethics Committee of Liuhe District People’s Hospital in Nanjing. RESULTS AND CONCLUSION: When osteoblasts were cultured to the fourth generation, followed by alkaline phosphatase staining, black grey granules were found in the cytoplasm of the positive cells, and the morphology of the cells was irregular. When the concentration of catalpol was lower than 1 mg/L, it had no significant effect on osteoblasts; when it was higher than 10 mg/L, it had a significant effect on the proliferation of osteoblasts but no drug toxicity (P < 0.05). When the concentration of lipopolysaccharide was higher than 80 mg/L, there was a significant trend of inflammatory damage to osteoblasts (P < 0.01). Therefore, 80 mg/L was selected as the best injury concentration in this experiment. When the concentration of catalpol from the root of Rehmannia glutinosa was lower than 1 mg/L, it had no significant protective effect on the inflammatory response of osteoblasts (P < 0.05); when it was higher than 10 mg/L, it had significant protective effect on the inflammatory response of osteoblasts (P < 0.05). Therefore, when the concentration of catalpol from the root of Rehmannia glutinosa reaches a certain level, it has no toxic effect on osteoblasts, promotes proliferation, and has anti-inflammatory protective effect on inflammatory osteoblasts induced by lipopolysaccharide. [ABSTRACT FROM AUTHOR]

In the current study, two similar substituted imidazo[1,2-α]pyridine ligands with different coordination groups were facilely synthesized, and were further used for construction of two novel Cu(II)-based coordination complexes with the chemical formulae of Cu(L1)2(H2O)4 (1, HL1 = (E)−4-(3-(2-chloroimidazo[1,2-a]pyridin-3-yl)acryloyl)benzoic acid) and [Cu(L2)2]n (2, HL2 = (E)−4-(2-chloroimidazo[1,2-α]pyridin-3-yl)−2-oxobut-3-enoic acid) via a one pot solvothermal reaction. In addition, their inhibitory effect on human myocardial aneurysm cells were evaluated. Firstly, to explore the inhibitory effect of complexes on cancer cell viability and proliferation, the Cell Counting Kit-8 (CCK-8) assay was performed. Next, to evaluate the influence of the compounds on the cell migration and invasion ability, the transwell assay was conducted. In addition to this, for the cancer cell apoptosis evaluation after compounds treatment, the flow cytometry was carried out in this research. [ABSTRACT FROM AUTHOR]

Under solvothermal conditions, two new coordination polymers (CPs), namely {[Cu(HL)]·2CH3CN}n (1) and {[Cu(HL)(bimb)]·4H2O}n (2) (H3L = tris(p-carboxyphenyl)phosphane oxide, bimb = 1,4-bis(imidazol-1-ylmethyl)benzene) were prepared. Their treatment activity on the human prostate cancer was evaluated through series of biological experiments. Firstly, the human prostate cancer cells proliferation was determined with Cell Counting Kit-8 (CCK-8) detection kit. The influence of compounds 1 and 2 on the cancer cells migration and invasion ability was measured with trans-well assay. Next, the important role of the EGF-R human prostate cancer cells was determined with western blotting. In the end, the activation of the HIPPO signaling pathway was detected with real-time reverse transcription polymerase chain reaction (RT-PCR). [ABSTRACT FROM AUTHOR]

Objective; To study the hepatotoxicity of Xianling Gubao extract. Methods; With different concentrations of Xianling Gubao extract and its unilateral extracts acting on human hepatocyte Hep3B, hepatotoxicity was detected by using CCK-8 kit. The cell model was validated by positive drug FCCP, and the dose effect reference value was thus obtained. ICso of different extracts was screened out, and the differences in hepatotoxicity were compared between various extracts. Results : The IC50 values of various extracts were as follows : Xianling Gubao extract was 153. 2 µg/ml, Epimedium brecicornu omission ex- tract was 312. 2 µg/ml, Dipsacus asper omission extract was 217. 1 µg/ml, Salvia miltiorrhiza omission extract was 256. 5 µg/ml, Anemarrhena asphodeloides omission extract was 229. 6 µg/ml, Psoralea corylifolia omission extract was 246. 9 µg/ml, Rehmannia glutinosa omission extract was 259. 4 µg/ml, Epimedium brevicornu extract was 91. 77 µg/ml, Dipsacus asper extract was 179. 0 µg/ml, Salvia miltiorrhiza extract was 410.6 µg/ml, Anemarrhena asphodeloides extract was 307. 9 µg/ml, Psoralea corylifolia extract was 73. 22 µg/ml and Rehmannia glutinosa extract was 180.2 µg/ml. The IC50 values of Xianling Gubao, Epimedium brecicornu and Psoralea corylifolia were significantly lower than those of other extracts. Therefore, it could be estimated that the substances that caused hepatotoxicity were mainly Epimedium brevicornu and Psoralea corylifolia. Conclusion : With CCK-8, the main hepatotoxic ingredients screened out in the formula of Xianling Gubao were chiefly Epimedium brecicornu and Psoralea corylifolia extracts. The research results could provide solid evidence for further study of the hepatotoxicity of Xianling Gubao. [ABSTRACT FROM AUTHOR]

The objective of this study was to evaluate the effect of combination of pheophorbide (Thyphonium flagelliforme) and andrographolide (Andrographis paniculata) on breast cancer cell culture of MCF-7 in a laboratory study using a posttest only control group design. human dermis fibroblast (HDF) cells were used as a control. Cell culture was obtained from YARSI University biorepository. Control and experimental treatment was given to six groups for each cell culture, namely control without treatment, P8 (pheophorbide 8 μM), A48 (andrographolide 48 μM), P2A12 (combination pheophorbide 2 μM and andrographolide 12 μM), P4A24 (combination pheophorbide 4 μM and andrographolide 24 μM), and P8A48 (pheophorbide combination 8 μM and andrographolide 48 μM). The treatment was given for 24 h, with three repetitions for each group. Results showed a decrease in cell viability in all treatment groups compared to the control group both in MCF-7 cells and HDF cells. Increasing the dose did not cause an increase in toxicity to MCF-7 cells. The use of a combination of pheophorbide and andrographolide showed anti-cancer effectiveness in MCF-7 breast cancer cell lines at 4 μM pheophorbide doses and 24 μM andrographolide [ABSTRACT FROM AUTHOR]

Acute pancreatitis is an inflammatory process of the pancreas and a leading cause of hospitalization amongst gastrointestinal disorders. Previously, cholecystokinin (CCK) has been described to play a role in regeneration of pancreas. The aim of this study was to analyse the function of cholecystokinin octapeptide (CCK-8) during induced pancreatitis in an animal model. Overall acute pancreatitis was induced in 38 pigs. After the induction of acute pancreatitis, half of the animals were treated with CCK-8. Intraoperative clinical data, postoperative blood parameters, 'Porcine Well-being' (PWB) and fitness score and post-mortal histopathological data were analysed. At baseline, physiologically parameters of the pigs of both groups were comparable. No differences were observed regarding the overall survival of animals (p = 0.97). Postoperative PWB score were significantly enhanced in animals treated with CCK-8 as compared to the control group (p = 0.029). Moreover, histopathological analysis of the pancreatic tissue revealed that acinar necrosis and edema were significant reduced in the CCK-8 group in comparison to the control group (p = 0.016 and p = 0.019). In conclusion, we found that CCK-8 treatment reduces acinar necrosis and edema of pancreatic tissue after induction of an acute pancreatitis in pigs. [ABSTRACT FROM AUTHOR]

Objective: The miR-638 acted as a tumor suppressor and E2F transcription factor 2 (E2F2) was a critical regulator in some cancers, while the role of them on stemness of breast cancer stem cells (BCSCs) was rarely detailed. Hence, we focused on exploring the effects of miR-638 and E2F2 on BCSCs stemness. Methods: The proportion of CD24 −/CD44 + cells of BCSCs was detected by flow cytometry. The target relationship of miR-638 and E2F2 was explored using luciferase assays. The ability of self-renewal, proliferation, and invasion of BCSCs were determined by Mammosphere forming, Cell Counting Kit-8 (CCK-8), colony formation, and transwell assays. Xenograft tumor was established to detect the influence of miR-638 on tumor growth. Results: miR-638 was down-regulated, while E2F2 was elevated in breast cancer. The E2F2 level was negatively correlated with miR-638. The BCSCs represented higher proportion of CD24 −/CD44 + cells and levels of sex determining region Y-box 2 (SOX2) and octamer-binding transcription factor 4 (OCT4). The miR-638 was down-regulated and E2F2 was increased in BCSCs. MiR-638 could target to E2F2 and decreased the level of E2F2 in BCSCs cells. Overexpression of miR-638 decreased the proportion of CD24 −/CD44 + cells and the levels of SOX2 and OCT4 by inhibiting E2F2. The overexpression of miR-638 also inhibited the abilities of self-renewal, proliferation, and invasion of BCSCs by inhibiting E2F2. The miR-638 overexpression inhibited the breast tumor growth. Conclusion: MiR-638 represses the characteristics and behaviors of BCSCs by targeting E2F2. MiR-638 may be a potential target for breast cancer therapy. [ABSTRACT FROM AUTHOR]

The first total synthesis of marine natural product, (−)-majusculoic acid (1) and its seven analogs (9–15), was accomplished in three to ten steps with a yield of 3% to 28%. The strategy featured the application of the conformational controlled establishment of the trans-cyclopropane and stereochemical controlled bromo-olefination or olefination by Horner–Wadsworth–Emmons (HWE) reaction. The potential anti-inflammatory activity of the eight compounds (1 and 9–15) was evaluated by determining the nitric oxide (NO) production in the lipopolysaccharide (LPS)-induced mouse macrophages RAW264.7. (−)-Majusculoic acid (1), methyl majusculoate (9), and (1R,2R)-2-((3E,5Z)-6-bromonona-3,5-dien-1-yl)cyclopropane-1-carboxylic acid (12) showed significant effect with inhibition rates of 33.68%, 35.75%, and 43.01%, respectively. Moreover, they did not show cytotoxicity against RAW264.7 cells, indicating that they might be potential anti-inflammatory agents.

A series of halogenated Phenstatin analogs were designed as microtubule destabilizing agent by docking study. It was synthesized within three steps starting from 2-chloro-5-iodobenzoic acid and substituted benzene. All the products were characterized by 1H NMR and 13C NMR spectral analysis, and the stereochemical structure was also confirmed by a single crystal X-ray diffraction crystallographic analysis. The microtubule destabilizing activities were evaluated in vitro with human liver cancer Huh-7 cell line and human lung cancer A549 cell line. Some of the HPAs were achieved IC50 about 5.0 μM against human liver cancer Huh-7 cells. [ABSTRACT FROM AUTHOR]

Background: Treating colorectal cancer (CRC) continues to be a clinical challenge. Studies have shown that epithelial-mesenchymal transition (EMT) is a critical step in tumor progression and transforming growth factor-β1 (TGF-β1) signaling has been shown to play a crucial role in EMT. Here, we investigate the inhibition effect of Ginsenoside Rb2, main bioactive component of ginseng, in human colorectal cancer cells via TGF-β1.Purpose: The current study aims to study the inhibitory effect of Ginsenoside Rb2 on HCT116 and SW620 cells and its anti-tumor mechanism.Methods: Histomorphological analysis and western blot analysis were performed to evaluate expression of TGF-β1 in human cancerous colon samples and the adjacent normal samples. The docking simulation assay were performed to explore the potential mode of binding of Ginsenoside Rb2 to the TGF-β1 protein. CCK8, adhesion and invasion assay were used to assess the effects of Ginsenoside Rb2 in HCT116 and SW620 cells. RT-PCR, Western blot and Immunohistochemical staining were employed to detect the TGF-β1-related signaling pathways in the colon cancer cells and/or xenograft mice.Results: The expression of TGF-β1 in human cancerous colon samples was significantly increased compared with the adjacent normal samples. Ginsenoside Rb2 inhibit the growth, adhesion, EMT and metastasis of human colorectal cancer cells. The docking simulation assay confirmed that Ginsenoside Rb2 bound to the hydrophobic pocket of TGF-β1, which partially overlaps with the binding sites on TGF-β1, and thus disrupted TGF-β1 dimerization. Western Blot analysis further confirmed that Ginsenoside Rb2 could inhibit the expression of TGF-β1 in vitro and in vivo. Furthermore, Ginsenoside Rb2 could inhibit the expression of Smad4 and phosphorylated Smad2/3.Conclusion: Ginsenoside Rb2 could inhibit EMT of colorectal cancer cells through the TGF-β1/Smad signaling, and might be a potential candidate for the treatment of colorectal cancer. [ABSTRACT FROM AUTHOR]

We hypothesize that cholecystokinin (CCK) and glucagon-like peptide-1 (GLP-1) are involved in the modulation of metabolic regulation of food intake by fatty acids in fish. Therefore, we assessed in rainbow trout (Oncorhynchus mykiss) the effects of intracerebroventricular treatment with 1 ng/g of CCK-8 and with 2 ng/g of GLP-1 on food intake, expression of neuropeptides involved in food intake control and the activity of fatty acid-sensing systems in hypothalamus and hindbrain. Food intake decreased up to 24 h post-treatment to 49.8-72.3% and 3.1-17.8% for CCK-8 and GLP-1, respectively. These anorectic responses are associated with changes in fatty acid metabolism and an activation of fatty acid-sensing mechanisms in the hypothalamus and hindbrain. These changes occurred in parallel with those in the expression of an orexigenic and orexigenic peptides. Moreover, we observed that the activation of fatty acid sensing and the enhanced anorectic potential elicited by CCK-8 and GLP-1 treatments occurred in parallel with the activation of mTOR and FoxO1 and the inhibition of AMP Kα, BSX and CREB. The results are discussed in the context of metabolic regulation of food intake in fish. [ABSTRACT FROM AUTHOR]

Objectives: Tooth-colored composites have emerged as a standard restorative material in caries therapy and have largely replaced materials such as silver amalgam or glass ionomer cements. In addition to their superior esthetics and desirable mechanical properties, composites also comprise negative characteristics, such as wear, shrinkage, and an adverse biocompatibility. Modifications of classic resin-based dental composites have been developed to overcome these shortcomings. For example, ormocers are innovative inorganic-organic hybrid polymers that form a siloxane network modified by the incorporation of organic groups. Recently, a new ormocer, Admira Fusion (VOCO), was introduced to composite technology. The absence of cytotoxic matrix monomers leads to the hypothesis that ormocers have improved biocompatibility compared to resin-based dental restorative materials.Materials and methods: The aim of this study was to compare the cytotoxic effects of Admira Fusion to a nanohybrid composite (GrandioSO, VOCO) and a nanofiller composite (Filtek Supreme XTE, 3M Espe) on the standard dermal mouse fibroblasts (L929) and human gingival fibroblasts (GF-1) via a Cell Counting Kit-8 (CCK-8) assay.Results: Admira Fusion was significantly less cytotoxic than GrandioSO and Filtek Supreme XTE to both the standard mouse dermal fibroblasts (L929) and human gingival fibroblasts.Conclusions: Compared to other resin-based dental restorative materials, the ormocer (Admira Fusion) possesses a superior biocompatibility in vitro. Future research studies are needed to confirm our results.Clinical significance: Clinically, dental practitioners and their patients might benefit from Admira Fusion in terms of reduced adverse biologic reactions compared to resin-based dental restorative materials. [ABSTRACT FROM AUTHOR]

BACKGROUND: The domestic porous tantalum implant material has a three-dimensional spatial structure that may affect the growth and secretion of cells. OBJECTIVE: To investigate the proliferation, cell cycle, osteogenic secretion of osteoblasts co-cultured with domestic porous tantalum scaffolds. METHODS: Passage 3 rabbit osteoblasts were extracted and divided into three groups: normal culture group, control group added with the porous tantalum extract, and experimental group co-cultured with the porous tantalum scaffold. The proliferation of cells in each group was detected by cell counting kit-8 method at 3, 5, 7 days of culture. The growth of osteoblasts on the porous tantalum scaffold was observed by scanning electron microscopy. ELISA kit assay was applied to detect the levels of osteocalcin and type I collagen in osteoblasts at 3, 5, 7 days of culture. The cell circle of osteoblasts was detected by flow cytometry at 7 days of culture. RESULTS AND CONCLUSION: (1) Cell proliferation: There was no difference in cell proliferation among the three groups at different time points (P > 0.05). (2) Scanning electron microscopy: On the 3rd day of compound culture, the cells adhered to the scaffold surface and pores, and arranged sparsely with few protrusions. On the 5th day, the cells began to extend, and the adjacent cells were interconnected to form a flaky growth. On the 7th day, the cells merged into pieces, and the secreted matrix covered most of the scaffold surface. (3) ELISA detection: With the cultivation time, the osteocalcin secretion from the osteoblasts was gradually increased in all the three groups, while the secretion of type I collagen was increased first and then decreased. The secretion of osteocalcin and type I collagen in the experimental group at different time points were higher than that in normal culture and control groups. (4) Cell cycle: The osteoblasts in the three groups were normal diploid cells, no aneuploid cells appeared, and the cell DNA content was normal. The cell cycle distribution was similar in the three groups. To conclude, the domestic porous tantalum scaffolds have good biocompatibility and may promote osteoblast mineralization and osteogenesis. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • A novel method for the synthesis of carbon dot-decorated materials was developed. • Both OH and O 2 − were produced by the CD/Na 2 W 4 O 13 /WO 3 composite. • The observed antibacterial activity enhancement was mainly attributed to the incorporation of carbon dots. • An environmentally friendly photocatalyst with antibacterial activity was developed. Abstract Photodisinfection by semiconductors has been proven to be an effective method for achieving antibacterial or antifungal activity. However, the toxicity of the nanomaterial to the environment and organisms is a major concern. Herein, a highly efficient and environmentally friendly photodisinfection material of a carbon dots (CDs) decorated Na 2 W 4 O 13 composite with WO 3 photocatalyst was fabricated via a facile hydrothermal-calcination approach. The TEM (transmission electron microscopy) images showed that CDs decorated the surface of the Na 2 W 4 O 13 flakes. Compared with the samples without incorporated CDs, the as-synthesized composite of CDs/Na 2 W 4 O 13 /WO 3 exhibited excellent antimicrobial activity against E. coli under visible light illumination. Electron spin resonance (ESR) spectroscopy and reactive species scavenging experiments revealed that the hydroxyl radicals and superoxide radical anions played the most important role in the photocatalytic bacterial inactivation. Furthermore, the cytotoxicity of the CDs/Na 2 W 4 O 13 /WO 3 composite was evaluated by analyzing the viability of HepG2 and Chinese hamster lung (V79) cells using Cell Counting Kit-8 (CCK-8). [ABSTRACT FROM AUTHOR]

In this study, sterlet (Adpenser ruthenus) was chosen as the model species of sturgeon, different solutions were used to isolate the sturgeon peripheral blood lymphocytes and study their optimal proliferate response condition. Phytohemagglutinin (PHA), concanavalin A (ConA) and lipopolysaccharide (LPS) were used as lymphocyte proliferation mitogen, respectively. According to L25 (56) five-factor five-level orthogonal experimental design, the conditions for sturgeons proliferation response of peripheral blood lymphocytes were optimized using enhanced cell counting Kit-8 (enhanced CCK-8 or WST-8). Five factors were selected to explore the optimal response conditions, including culture time, culture temperature, ceU concentration, fetal bovine serum (FBS) concentration, and mitogen concentration. The results showed that 70V Percoll (1. 092 g/ml) used as the sturgeon lymphocyte separation solution had the best separating effect. The optimal proliferation conditions were as follows: 3.625 X 106 initial ceüs, 20 μg m l PHA or 50 μg m l ConA or 10 μg/ml LPS a mitogen, 10V - 20V FBS, the temperature at 20 - 25°C, and the culture time of 2 d. [ABSTRACT FROM AUTHOR]

Objective To investigate the effect of ginsenoside Rg3 on mouse neural stem cells (NSCs) proliferation in vitro．Methods Fetal cortices of embryonic 14d (E14) C57BL/6 mouse were isolated for culturing primary NSCs．After being passaged twice， NSCs specific antibodies， Nestin and Sox2 were identified by immunocyto chemistry staining. CCK-8 assay and BrdU labelling assay were used to measure NSCs proliferation in response to different ginsenoside Rg3 concentrations (20 nM，50 nM，250 nMand 500 nM) . Results Primary and passaged NSCs were successfully cultured and almost all the cells were both Nestin and Sox2 positive NSCs， which indicated high purity of NSCs. BrdU labelling assay showed the group of 50 nM Rg3 had the highest relative ratio of BrdU positive percentage， and the difference with other groups was statistically significant (P<0.01) ． CCK-8 assay also showed 50 nM Rg3 group had the highest OD value， which was significantly higher than the control group (P<0.01)，20 nMgroup (P<0.05) and 500 nMgroup (P<0.01)， while other groups had no significant difference between each other(P>0.05) . Conclusion 50 nM ginsenoside Rg3 could significantly promote mouse NSCs proliferation in vitro. [ABSTRACT FROM AUTHOR]

For the development of potential anti-prostate cancer agents, 24 kinds of novel naftopidil-based arylpiperazine derivatives have been synthesized and characterized by spectroscopic methods. Their antitumor activities were evaluated against several classical prostate cancer cell lines including PC-3, LNCaP, and DU145. Among all the compounds, 9 , 13 , 17 , 21 and 27 showed strong cytotoxic activities against DU145 cells (IC 50  < 1 μM). Further testing confirmed that compound 17 inhibited the growth of DU145 cells by inducing cell cycle arrest at G0/G1 phase. Besides, antagonistic activities of compounds ( 9 , 13 , 17 , 21 and 27 ) towards a 1 -ARs (α 1A , α 1B , and α 1D ) were further evaluated using dual-luciferase reporter assays, and the compounds 13 and 17 exhibited better a 1 -ARs subtype selectivity. The structure–activity relationship (SAR) of these developed arylpiperazine derivatives was rationally discussed. Taken together, these results suggested that further development of such compounds may be of great interest. [ABSTRACT FROM AUTHOR]