Disruption of O-GlcNAcylation Homeostasis Induced Ovarian Granulosa Cell Injury in Bovine.

O-linked ß-N-acetylglucosamine (O-GlcNAc) modification is a ubiquitous, reversible, and highly dynamic post-translational modification, which takes charge of almost all biological processes examined. However, little information is available regarding the molecular regulation of O-GlcNAcylation in granulosa cell function and glucose metabolism. This study focused on the impact of disrupted O-GlcNAc cycling on the proliferation and apoptosis of bovine granulosa cells, and further aimed to determine how this influenced glucose metabolism. Pharmacological inhibition of OGT with benzyl-2-acetamido-2-deoxy-α-D-galactopyranoside (BADGP) led to decreased cellular O-GlcNAc levels, as well as OGT and OGA protein expressions, whereas increasing O-GlcNAc levels with the OGA inhibitor, O-(2-acetamido-2-deoxy-D-gluco-pyranosylidene) (PUGNAc), resulted in elevated OGA protein expression and decreased OGT protein expression in granulosa cells. Dysregulated O-GlcNAc cycling reduced cell viability, downregulated the proliferation-related genes of CDC42 and PCNA transcripts, upregulated the pro-apoptotic genes of BAX and CASPASE-3 mRNA and the ratio of BAX/BCL-2, and increased the apoptotic rate. Glycolytic enzyme activities of hexokinase and pyruvate kinase, metabolite contents of pyruvate and lactate, mitochondrial membrane potential, ATP levels, and intermediate metabolic enzyme activities of succinate dehydrogenase and malate dehydrogenase involved in the tricarboxylic acid cycle, were significantly impaired in response to altered O-GlcNAc levels. Moreover, inhibition of OGT significantly increased the expression level of thioredoxin-interacting protein (TXNIP), but repression of OGA had no effect. Collectively, our results suggest that perturbation of O-GlcNAc cycling has a profound effect on granulosa cell function and glucose metabolism.

[The anti-tumor activity and molecular mechanisms of an Aurora kinase inhibitor ZLJ213 in suppressing colon cancer growth].

The aim of this study is to evaluate anti-tumor activities and mechanism of a novel kinase inhibitor ZLJ213 which targeted Aurora A and vascular endothelial growth factor receptor (VEGFR) in vitro and in vivo against human colon cancer. Results showed that ZLJ213 inhibited cell proliferation and induced cell cycle arrest and apoptosis of HCT1 16 and SW48 cell lines. In HCT116-derived xenograft, ZLJ213 dosed at 100 mg · kg(-1) inhibited tumor growth by 73.24%. The IC50 of ZLJ213 on the expression of p-Aurora A was 0.258 µmol · L(-1) analyzed by ELISA. Under the concentration of 0.08 µmol · L(-1), ZLJ213 could inhibit the activities of Aurora A, Histone H3 and VEGFR of HCT116 and SW48 cell lines. Simultaneously, ZLJ213 induced activation of Caspase 3 and PARP cleavage. Above data suggested that ZLJ213 had the ability to inhibit cell proliferation and induce cell apoptosis both in vitro and in vivo in colon cancer, and down-regulate the expression of p-Aurora A and p-VEGFR. ZLJ213 might be a potential therapeutic agent against colon cancer.

[Synthesis of novel curcumin mimics and preliminary evaluation for their antitumor activity].

Curcumin has been reported to possess antitumor activity with low toxicity. However, the clinical application of curcumin has been significantly limited by its instability and poor metabolic property. In order to overcome these limitations and discover novel small molecules with potential antitumor activity, 29 curcumin mimics were synthesized, which were confirmed by 1H NMR and HR-MS, and their cytotoxic property was evaluated against five human cancer cell lines in vitro. Compounds 16, 18 and 19 exhibited good cytotoxic property, their IC50 value were even below 5 micromol x L(-1) to some cancer cell lines, 5-9 times better than curcumin.

[Synthesis and in vitro cytotoxic activities of sorafenib derivatives].

A series of novel sorafenib analogues were designed and synthesized. The cytotoxic activities of these compounds were tested in four tumor cell lines. Some of the compounds showed potent antiproliferative activity against the tested cell lines with IC50 = 4-20 micromol x L(-1). Some compounds demonstrated competitive antiproliferative activities to sorafenib against tested cancer cell lines. Among them, compound 7c demonstrated significant inhibitory activities on ACHN, HCT116 and MDA-MB-231 cell lines with IC50 values of 9.01, 4.97, 6.61 micromol x L(-1), respectively.

[Anti-tumor activity and mechanism of T03 in vitro and in vivo].

The purpose of this study is to investigate the activity and mechanism of a new anti-tumor agent T03. MTT and colony formation assay were performed to determine anti-proliferation activity of T03 in vitro. Antitumor activity was observed by Renca xenograft model in vivo. The effect of T03 on cell cycle and apoptosis were measured by FCM analysis. Western blotting was performed to investigate the expression level of proteins in HepG2 cell lines treated with T03. T03 had anti-tumor activity by inhibiting tumor cell growth and colony formation in vitro, especially on hepatocellular carcinoma cells (HCC). At the concentration of 10 micromol x L(-1), T03 induced cell apoptosis and cell cycle arrest in HCC. Moreover, it proved that T03 reduced the tumor weight with the rate of 42.30% without any obviously side effect in Renca xenograft model. At the concentration of 2.0 micromol x L(-1), T03 was able to reduce the level of p-c-Raf (Ser259), and thus blocked Raf/MEK/ERK and AKT signaling in HepG2 cell lines. The result suggested that T03 has the potential to inhibit cell proliferation and induce cell apoptosis both in vitro and in vivo, particularly active against HCC, indicating T03 and its analogues may serve as a new anti-cancer drug against hepatocellular carcinoma.

[Synthesis and biological evaluation of 2-(3-butynoicamidophenyl) benzothiazole derivatives as antitumor agents].

A series of 2-(3-butynoicamidophenyl)benzothiazole derivatives were synthesized starting from 4-fluoro-3-nitrobenzoic acid. Structures of all the synthesized compounds were confirmed by 1H NMR and HR-MS. Their antitumor activities against human tumor cells lines (HCT116, Mia-PaCa2, U87-MG, A549, NCI-H1975) were evaluated by MTT assay. The results revealed that most of the synthesized compounds showed potent activities against HCT116, Mia-PaCa2, U87-MG tumor cells lines. Particularly, compounds 14c and 14h exhibited better activity with IC50 values of 1 x 10(-8) mol x L(-1) against U87-MG and HCT116 respectively. The structure-activity relationship of compounds was also discussed preliminarily.

[Establishment and application of screening methods for non-agonist PPARγ ligand].

In-vitro assay methods were established to evaluate transactivation and binding activity of compounds on peroxisome proliferator-activated receptor y (PPARγ). Firstly, plasmids were constructed for transactivation assay of PPARγ response element (PPRE) triggered reporter gene expression, and for cell-based binding activity assay of the chimeric receptor, which was fused with PPARγ ligand binding domain (LBD) and yeast transcriptional activator Gal4. Secondly, by using PPARy competitive binding assay based on time resolved-fluorescence resonance energy transfer (TR-FRET), affinities of compounds and drugs to PPARγ were evaluated. In application of these above methods, the PPARγ activating potency and characteristics of different compounds were evaluated, and a novel benzeneselfonamide derivative, ZLJ01, was found to have comparable binding activity and affinity with the well-known PPARy agonist, but lack of PPRE mediated transactivation activity. In preliminary study on in-vitro hypoglycemic activity, ZLJ1 was found to promote insulin-stimulated glucose uptake by liver cells. Therefore, we believe that combining transactivation and binding activity as well as affinity evaluation, the system could be used to screen non-agonist PPARγ ligand as anovel PPARγ modulator

Andrographolide sensitizes KRAS-mutant colorectal cancer cells to cetuximab by inhibiting the EGFR/AKT and PDGFRß/AKT signaling pathways.

BACKGROUND: Cetuximab, an inhibitor targeting EGFR, is widely applied in clinical management of colorectal cancer (CRC). Nevertheless, drug resistance induced by KRAS-mutations limits cetuximab's anti-cancer effectiveness. Furthermore, the persistent activation of EGFR-independent AKT is another significant factor in cetuximab resistance. Nevertheless, the mechanism that EGFR-independent AKT drives cetuximab resistance remains unclear. Thus, highlighting the need to optimize therapies to overcome cetuximab resistance and also to explore the underlying mechanism. PURPOSE: This work aimed to investigate whether and how andrographolide enhance the therapeutic efficacy of cetuximab in KRAS-mutant CRC cells by modulating AKT. METHODS: The viabilities of CRC cell lines were analyzed by CCK-8. The intracellular proteins phosphorylation levels were investigated by Human Phospho-kinase Antibody Array analysis. Knockdown and transfection of PDGFRß were used to evaluate the role of andrographolide on PDGFRß. The western blotting was used to investigate Wnt/ß-catenin pathways, PI3K/AKT, and EMT in KRAS-mutant CRC cells. The animal models including subcutaneous tumor and lung metastasis were performed to assess tumor response to therapy in vivo. RESULTS: Andrographolide was demonstrated to decrease the expression of PI3K and AKT through targeting PDGFRß and EGFR, and it enhanced cetuximab effect on KRAS-mutant CRC cells by this mechanism. Meanwhile, andrographolide helped cetuximab to inhibit Wnt/ß-catenin, CRC cell migration and reduced Vimentin expression, while increasing that of E-cadherin. Lastly, co-treatment with cetuximab and andrographolide reduced the growth of KRAS-mutant tumors and pulmonary metastases in vivo. CONCLUSIONS: Our findings suggest that andrographolide can overcome the KRAS-mutant CRC cells' resistance to cetuximab through inhibiting the EGFR/PI3K/AKT and PDGFRß /AKT signaling pathways. This research provided a possible theory that andrographolide sensitizes KRAS-mutant tumor to EGFR TKI.

Zygomatic surface marker-assisted surgical navigation: a new computer-assisted navigation method for accurate treatment of delayed zygomatic fractures.

PURPOSE: To describe a new method of zygomatic surface marker navigation to treat delayed unilateral zygomatic fractures. PATIENTS AND METHODS: The computed tomography (CT) data for 6 patients were obtained before surgery and imported into the surgical planning software. After 3-dimensional (3D) construction and segmentation, 3D cylindrical-shaped objects in stereolithographic format were placed in position and merged with the data from the fractured segments to mark the area for surface reduction. Data from the unaffected side were used to guide the reduction data for the segments with markers. During surgery, the surface markers were marked by drilling holes in the fractured bones in a process guided by the surgical navigation plan established before osteotomy. The segments were then reduced to the predetermined places using the positions of the hole markers as guides. 3D image comparisons and axial CT measurements were used to evaluate navigation accuracy and bone symmetry. RESULTS: Six patients with unilateral delayed zygomatic fractures were treated using this approach. The mean deviation between the postoperative 3D images and the reduction navigation plan for the 6 patients was +1.24 mm and -1.4 mm. The mean width deviation between the affected and unaffected sides was 1.28 mm, and the mean eminence deviation was 1.22 mm. All patients were followed up for at least 3 months and experienced no obvious complications. CONCLUSIONS: Zygomatic surface marker-assisted surgical navigation can simplify the navigation planning for surgery and avoid the complex protocols needed to create the surgical templates. The navigation accuracy was acceptable, and all 6 patients obtained good facial symmetry.

Pd(II) and Zn(II) based complexes with Schiff base ligands: synthesis, characterization, luminescence, and antibacterial and catalytic activities.

Two new metal complexes involving Schiff base ligands, namely, [Pd(L1)2] (1) and [Zn(L2)2] (2), [HL1: 2,4-dibromo-6-((E)-(mesitylimino)methyl)phenol and HL2: 2-((E)-(2,6-diisopropylphenylimino)methyl)-4,6-dibromophenol], have been solvothermally synthesized and characterized by elemental analysis, IR-spectroscopy, thermogravimetric analysis, powder X-ray diffraction, and single-crystal X-ray diffraction. Both 1 and 2 are mononuclear cyclometalated complexes with square planar and tetrahedral coordination geometry, respectively. 1 and 2 display photoluminescence in the solid state at 298 K (fluorescence lifetimes τ = 5.521 µs at 508 nm for 1; τ = 3.697 µs at 506 nm for 2). These Schiff base ligands and their metal complexes have been screened for antibacterial activity against several bacteria strains, and the results are compared with the activity of penicillin. Moreover, the Suzuki reaction of 4-bromoanisole with phenylboronic acid by 1 has also been studied.

[Salicylic acid derivatives as simplified and novel GK small molecule activators].

Glucokinase (GK) is a new target for the treatment of type II diabetes mellitus (T2DM). In order to find a structure-simplified small molecule GK activator, 19 salicylic acid derivatives were designed and synthesized based on new lead compound (1). Experimental results showed that the potency of compound 8h is superior to control RO-28-0450 in GK activation.

[Histone deacetylase inhibits the growth and migration of human osteosarcoma cells].

OBJECTIVE: To explore the effects of histone deacetylase inhibitors (HDACIs) on human osteosarcoma in vitro and in vivo. METHODS: HDACI suberoylanilide hydroxamic acid (SAHA) and sodium butyrate (SB) were employed to examine the effects on human osteosarcoma in vitro and in vivo. The in vitro effects of HDACI SAHA and SB were evaluated in SaOS2 and U2OS human osteosarcoma cell lines. Cell growth, cell cycle progression and histone acetylation were examined by methyl tolyl sulfide (MTS), flow cytometry and Western blot respectively.In addition, SAHA or SB was administered for 4 weeks in a murine xenograph model for assessing the in vitro effects. RESULTS: MTS assay revealed that SAHA and SB significantly suppressed the growth of SaOS2 and U2OS cells in a concentration-dependent manner.Western blot analysis indicated that the levels of acetylated H3 increased after HDACI treatment.Flow cytometry showed that SAHA arrested the cell cycle in G1 and G2/M phase, while SB arrested the cell cycle in G2/M phase. The tumor growth of murine xenograph model with SaOS2 was inhibited by SAHA and SB compared with vehicle control. CONCLUSION: HDACI SAHA and SB significantly inhibit the growth of human osteosarcoma cells and induce cell cycle arrest. Meanwhile, the tumor inhibitory effects were also validated in a murine xenograft model.

Numerical simulation of mechanical tests on a living skin using anisotropic hyperelastic law.

The skin is a living tissue that behaves in a hyperelastic and anisotropic way. A constitutive law called HGO-Yeoh is proposed to model the skin by improving the classical HGO constitutive law. This model is implemented in a finite element code FER "Finite Element Research" to benefit from its tools, including the bipotential contact method, a very efficient function coupling contact and friction. Identifying the skin-related material parameters is done through an optimisation procedure using analytic and experimental data. A tensile test is simulated using the codes FER and ANSYS. Then, the results are compared with the experimental data. Finally, a simulation of an indentation test using a bipotential contact law is done.

[Design, synthesis and antitumor activity of sorafenib analogues containing 2-picolinylhydrazide moiety].

A novel series of sorafenib analogs containing 2-picolinyl hydrazide moiety were designed and synthesized. In vitro, most of synthesized compounds have antiproliferation activity on MDA-MB-231, ACHN, HepG2, Mia-PaCa-2 and SW1990 cell lines tested by MTT assay. It is worth noting that the antitumor activities of compounds 2c, 2d and 2f are more potent than that of sorafenib on pancreatic cancer cells Mia-PaCa-2 and SW1990, and the activities of compounds 3f and 3g are 2-3 times than that of sorafenib on human hepatocellular carcinoma HepG2 cell line.

3-Methyl-benzene-1,2-diamine.

The title compound, C7H10N2, was synthesized from 2-methyl-6-nitro-aniline by a reduction reaction. In the crystal, molecules are linked via N-H⋯N hydrogen bonds, forming two-dimensional networks lying parallel to (100). These networks are stabilized by C-H⋯π and N-H⋯π inter-actions.

2,6-Dichloro-N-(4-methyl-phen-yl)benzamide.

In the title compound, C(14)H(11)Cl(2)NO, the two benzene rings are non-coplanar [dihedral angle = 60.9 (3)°]. In the crystal, an amide N-H⋯O hydrogen bond links the mol-ecules into chains which extend along (001).

[A comparison of efficacies between transjugular intrahepatic portosystemic shunt versus portoazygos devascularization in the treatment of portal hypertension with variceal bleeding].

OBJECTIVE: To compare the efficacies between transjugular intrahepatic portosystemic shunt (TIPS) and portoazygos devascularization (PAD) in the treatment of portal hypertension with variceal bleeding. METHODS: From December 1993 to December 2010, 309 patients with portal hypertension and variceal bleeding were admitted. According to their general conditions and Child-Pugh grades, they were assigned to undergo TIPS (group A, n = 235) or PAD (group B, n = 74). Before operation, compared with the PAD group, the TIPS group possessed worse liver functions, more severe ascites and a greater frequency of bleeding. After operation, the therapeutic efficacies and changes of portal hemodynamics, recurrent variceal bleeding, post-operative encephalopathy and long-term survival were evaluated between two groups. RESULTS: The postoperative portal pressure in the TIPS group ((42.6 ± 7.0) vs (26.3 ± 4.1) cm H2O) decreased much more than that in the PAD group ((38.7 ± 5.2) vs (33.5 ± 5.8) cm H2O, P < 0.01). The rebleeding rates during early postoperation were 0.85% (2/235) and 6.76% (5/74) in TIPS and PAD groups respectively, the occurring rates of hepatic encephalopathy 4.68% (11/235) and 4.05% (3/74) and the rates of operative mortality 1.70% (4/235) and 6.76% (5/74) respectively. Survival rates of 1, 3, 5 and 10 years were 98.30% (231/235) vs 92.24% (69/74), 92.41% (146/158) vs 88.06% (59/67), 80.77% (84/104) vs 79.25% (42/53), 51.43% (36/79) vs 51.85% (14/27) in TIPS and PAD groups respectively. CONCLUSIONS: As compared with PAD, TIPS offers the such advantages as less trauma, wider indication, faster hemostasis and satisfactory therapeutic efficacies. Especially for the emergency treatment of a patient with massive variceal bleeding and Child-Pugh C grade liver function, TIPS is a better option than PAD.

Enhanced segmentation of gastrointestinal polyps from capsule endoscopy images with artifacts using ensemble learning.

BACKGROUND: Endoscopy artifacts are widespread in real capsule endoscopy (CE) images but not in high-quality standard datasets. AIM: To improve the segmentation performance of polyps from CE images with artifacts based on ensemble learning. METHODS: We collected 277 polyp images with CE artifacts from 5760 h of videos from 480 patients at Guangzhou First People's Hospital from January 2016 to December 2019. Two public high-quality standard external datasets were retrieved and used for the comparison experiments. For each dataset, we randomly segmented the data into training, validation, and testing sets for model training, selection, and testing. We compared the performance of the base models and the ensemble model in segmenting polyps from images with artifacts. RESULTS: The performance of the semantic segmentation model was affected by artifacts in the sample images, which also affected the results of polyp detection by CE using a single model. The evaluation based on real datasets with artifacts and standard datasets showed that the ensemble model of all state-of-the-art models performed better than the best corresponding base learner on the real dataset with artifacts. Compared with the corresponding optimal base learners, the intersection over union (IoU) and dice of the ensemble learning model increased to different degrees, ranging from 0.08% to 7.01% and 0.61% to 4.93%, respectively. Moreover, in the standard datasets without artifacts, most of the ensemble models were slightly better than the base learner, as demonstrated by the IoU and dice increases ranging from -0.28% to 1.20% and -0.61% to 0.76%, respectively. CONCLUSION: Ensemble learning can improve the segmentation accuracy of polyps from CE images with artifacts. Our results demonstrated an improvement in the detection rate of polyps with interference from artifacts.

Bis(4-amino-2-chloro-phen-yl) disulfide.

The title compound, C(12)H(10)Cl(2)N(2)S(2), features an S-S bond [2.0671 (16) Å] that bridges two 4-amino-2-chloro-phenyl rings with a C-S-S-C torsion angle of -84.2 (2)°. The two benzene rings are twisted with respect to each other at a dihedral angle of 39.9 (2)°. Inter-molecular N-H⋯S hydrogen bonding is present in the crystal structure.

Bis(3,5-dimeth-oxy-phen-yl)phosphinic acid.

In the crystal structure of the title compound, C(16)H(19)O(6)P, inter-molecular O-H⋯O inter-actions link the mol-ecules into chains parallel to the b axis. These chains are linked by C-H⋯π and π-π inter-actions [centroid-centroid distance = 3.7307 (29) Å] into a three-dimensional network. The dihedral angle between the benzene rings is 73.5 (1)°. The C and O atoms of all four methoxy groups lie very close to the mean planes of their attached rings; the C atoms are 0.055 (2)-0.1038 (1) Šout of the mean plane of the attached rings.