Preparation, Optimization, and Characterization of Bovine Bone Gelatin/Sodium Carboxymethyl Cellulose Nanoemulsion Containing Thymol.

The aim of this study is to produce a biodegradable food packaging material that reduces environmental pollution and protects food safety. The effects of total solids content, substrate ratio, polyphenol content, and magnetic stirring time on bovine bone gelatin/sodium carboxymethylcellulose nanoemulsion (BBG/SCMC-NE) were investigated using particle size, PDI, turbidity, rheological properties, and zeta potential as evaluation indexes. The micro, structural, antioxidant, encapsulation, and release properties were characterized after deriving its optimal preparation process. The results showed that the nanoemulsion was optimally prepared with a total solids content of 2%, a substrate ratio of 9:1, a polyphenol content of 0.2%, and a magnetic stirring time of 60 min. SEM showed that the nanoemulsion showed a dense and uniform reticulated structure. FTIR and XRD results showed that covalent cross-linking of proteins and polysaccharides altered the structure of gelatin molecular chains to a more compact form but did not change its semi-crystalline structure. DSC showed that the 9:1 BBG/SCMC-NE had a higher thermal denaturation temperature and greater thermal stability, and its DPPH scavenging rate could reach 79.25% and encapsulation rate up to 90.88%, with excellent slow-release performance. The results of the study provide basic guidance for the preparation of stable active food packaging with excellent properties.

LINC00958/miR-627 signal axis regulates the proliferation, migration, and invasion of thyroid papillary carcinoma cells by TRIM44.

Papillary thyroid cancer (PTC) has attracted much attention due to its high morbidity and severe metastasis. Long noncoding RNA ENST00000504230 (LncRNA ENST00000504230, known as LINC00958) was overexpressed in many cancers and associated with cancer development. However, its underlying mechanism in PTC remains unclear. PTC tissues and corresponding adjacent tissues were collected for measuring the expression of LINC00958 and miR-627. MiR-627 and TRIM44 expressions were measured in in vitro cultured PTC cell lines (B-cpap and IHH4 cells) transfected with sh-LINC00958 or miR-627 mimic using RT-qPCR and western blot. Cell proliferation, migration, and invasion were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) and Transwell assays, respectively. Dual-luciferase reporter assay was performed to evaluate the target association between miR-627 and TRIM44. LINC00958 was up-regulated in PTC tissues and cells, while the expression of miR-627 was lowly expressed. Knockdown of LINC00958 inhibited the proliferation, migration, and invasion by elevating miR-627 expression in PTC cells. TRIM44 was confirmed as a target of miR-627. Overexpression of miR-627 in PTC inhibited the proliferation, migration, and invasion by down-regulating the expression of TRIM44. LINC00958 promoted proliferation, migration, and invasion in PTC by down-regulating miR-627 and activating TRIM44, indicating the potential therapeutic effect of LINC00958 on PTC.

Dihydroartemisinin induces pyroptosis by promoting the AIM2/caspase-3/DFNA5 axis in breast cancer cells.

BACKGROUND: Breast cancer is a complex disease. Recent research has examined the anticancer effects of dihydroartemisinin (DHA) on breast cancer. However, the molecular mechanism of the antitumour effect of DHA is unclear. METHODS: MCF-7 and MDA-MB-231 cell lines were used for in vitro research. BALB/c nude mice were used to establish breast cancer xenografts. The mRNA and protein levels were analysed by qRT-PCR and western blotting, respectively. Flow cytometry was performed to examine cell apoptosis. ELISA kits were used to evaluate the production of interleukin-1ß (IL-1ß) and IL-18. LDH and ATP release were individually measured with the corresponding kits. A colony formation assay was used to examine the proliferation of breast cancer cells. RESULTS: DHA inhibited proliferation and induced pyroptosis in breast cancer cells. Mechanistically, DHA activated the expression of absent in melanoma 2 (AIM2), caspase-3 and gasdermin E (DFNA5). In addition, AIM2 promoted DFNA5 expression by activating caspase-3. Knockdown of AIM2 and DFNA5 significantly enhanced breast cancer cell resistance to DHA. In vivo experiments showed that the tumorigenicity of breast cancer cells was significantly suppressed by DHA. Moreover, the AIM2/caspase-3/DFNA5 axis was activated by DHA and then induced pyroptosis. CONCLUSIONS: Our findings indicate that DHA inhibits tumorigenesis by inducing pyroptosis in breast cancer cells, highlighting a promising therapeutic strategy for breast cancer.

Combinational Treatment of Doxorubicin With Neoadjuvant Docetaxel for Different Subtypes of Patients With Breast Cancer.

AIM: The aim of this study is to characterize the effect of chemotherapy drug doxorubicin with neoadjuvant drug docetaxel for different molecular subtypes. METHODS: A total of 83 patients with late-stage breast cancer were chosen to undergo treatment and compared to these patients to the combinational treatment to identify the molecular characteristics that can predict the responses. RESULTS: Total response rate is 81.9% (68/83 patients). Among them, 7 patients show pathological complete response of 8.4%, 12 patients show clinical complete response of 14.5%, 49 patients show partial response of 59%, and 15 patients show stable disease of 18.1%. The comparison among different subtypes of breast cancer, including luminal A, luminal B, basal-like, and ERBB2+ subtypes, did not show statistical significant differences to the treatment of combinational treatment for the complete response rate, including pathological complete response and clinical complete response. Comparing with luminal A and luminal B subtypes, the ERBB2+ and basal-like subtypes have better complete response and response rate rates. The disease-free survival rate and overall survival rate at 29 months after treatment did not show statistical significant differences among different subtypes of patients with breast cancer. CONCLUSION: The molecular subtypes of breast cancer can predict responses to the combinational treatment of doxorubicin with docetaxel, and ERBB2+ and basal-like subtypes have better response rate and complete response rate. There is correlation of estrogen receptor and KI-67 level changes with response rate as well, where KI-67 high patients are more sensitive to the treatment.

Pinitol suppresses TNF-α-induced chondrocyte senescence.

Osteoarthritis (OA) is a highly prevalent joint disorder that is tightly correlated with age. As the body ages, cell replication and function decline until homeostasis can no longer be maintained. This process involves cellular senescence as well as replicative senescence. Telomere length, cell cycle arrest, expression of p16 and p53, and the release of senescence-associated ß-galactosidase (SA-ß-Gal) are all markers of cell senescence. In OA joints, chondrocytes undergo cellular senescence prematurely, thereby ceasing to synthesize and maintain cartilage tissue. Upregulation of proinflammatory cytokines, such as tumor necrosis factor-α (TNF-α), and oxidative stress induced by overproduction of reactive oxygen species (ROS) are key events in the pathogenesis of OA. In the present study, we investigated the effects of pinitol, a naturally occurring compound, on the effects of TNF-α on chondrocyte senescence and cell cycle arrest. We found that pinitol has a favorable safety profile in terms of cell viability. Pinitol significantly inhibited cellular senescence and cell cycle arrest in the G0/G1 phase induced by TNF-α. We also found that pinitol could inhibit TNF-α-induced increased telomerase activity and expression of p16 and p53. Importantly, we found that the effects of pinitol may be mediated through rescue of Nrf2 signaling, which is recognized as a key protective factor in OA. This finding was verified through a Nrf2 silencing experiment using Nrf2 siRNA. Together, our findings reveal the potential of pinitol as a safe therapeutic option for the prevention of OA-associated chondrocyte senescence and oxidative stress.

Molecular characterization of three GIBBERELLIN-INSENSITIVE DWARF1 homologous genes in hexaploid wheat.

GIBBERELLIN-INSENSITIVE DWARF1 (GID1) functions as a gibberellin (GA) receptor and is a key component in the GA signaling pathway. In this paper, three TaGID1 genes, orthologous to rice OsGID1 (the first identified GA receptor GID1 gene), were isolated from hexaploid wheat using homology cloning. Like OsGID1, the three homologous TaGID1 genes consisted of two exons and one intron. Physical location analyses using nullisomic-tetrasomic and deletion lines derived from the wheat cultivar Chinese Spring revealed that the three homologous TaGID1 genes reside in the chromosome bins 1AL3-0.61-1, 1BL1-0.47-0.69, and 1DL2-0.41-1. Accordingly, they were named TaGID1-A1, TaGID1-B1, and TaGID1-D1, respectively. The expression patterns of the three TaGID1 genes were determined by real-time PCR in various wheat tissues at the heading stage, including flag leaves, young spikes, peduncles, and the third and fourth internodes. The three TaGID1 genes had similar transcript patterns, and all exhibited greater expression in flag leaves than in the other tissues. Moreover, they were all down-regulated after treatment with exogenous gibberellic acid (GA(3)) in young seedlings, suggesting a feedback regulation of TaGID1 in wheat. Yeast two-hybrid assays demonstrated strong interactions between each putative TaGID1 and the wheat DELLA proteins RHT-A1, RHT-B1, and RHT-D1 in the presence of GA(3), and weak interactions in the absence of GA(3) in yeast cells. Furthermore, over-expression of each TaGID1 gene in the Arabidopsis double mutant gid1a/1c partially rescued the dwarf phenotype. These results suggest that the three TaGID1 homologous genes are all functional GA receptor genes in wheat.

Alkaline inulinase production by a newly isolated bacterium Marinimicrobium sp. LS-A18 and inulin hydrolysis by the enzyme.

To date, all of microbial inulinases reported showed optimal activity at pH values ranging from 3.5 to 7.0. A bacterial strain, Marinimicrobium sp. LS-A18, showing high extracellular inulinolytic activity was isolated from a marine solar saltern of the Yellow Sea in China. Maximum enzyme activity was obtained at 55°C and pH 9.0, respectively. The inulinase activity was induced by inulin, but not by the other carbon sources employed. Under the optimal medium and culture condition, the highest inulinase activity, 14.6 U/ml, was obtained after 96 h of incubation at shake flask level. The optimal medium for inulinase production was MHI medium containing 4% inulin, 1% peptone and 5% NaCl, while the optimal culture condition for inulinase production were pH 7.5, temperature 37°C, agitation speed 210 rpm, medium volume 40 ml in 250 ml shake flask, and incubation time 96 h. A large amount of monosaccharides was released after inulin hydrolysis by the inulinase from strain LS-A18. This is the first report on alkaline inulinase production from microorganism.