In this paper, a novel pH and redox dual-sensitive nanocarrier loaded with curcumin (Cur) and anticancer polypeptide (AP) was developed for dual targeting mitochondrial and CD44 receptor. The amphiphilic block copolymer was prepared by triphenylphosphonium (TPP)/oligomeric hyaluronic acid (oHA)/disulfide-menthone 1,2-glycerol ketal (SM), hereinafter referred to as TPP-oHSM. The TPP targeted the mitochondria, pH/redox dual-sensitive SM served as a hydrophobic part, and the CD44 receptor targeting oHA worked as a hydrophilic part. The chemical structure of the TPP-oHSM was identified using 1H NMR and FTIR technologies. Cur and AP were loaded into the TPP-oHSM micelles by self-assembly and denoted as C/A@TM. The C/A@TM prepared in this study exhibited an approximately spherical structure, with a mean diameter of 191.3 ± 3.1 nm and a negative zeta potential of -26.10 ± 0.45 mV. The in vitro release study and cellular uptake test revealed that the C/A@TM targeted the mitochondria and CD44 receptor, as well as it showed sensitivity towards pH and redox. In addition, the C/A@TM demonstrated satisfactory cytotoxic effects against MDA-MB-231 cells and MCF-7 cells. Finally, the in vivo application of the C/A@TM showed excellent therapeutic effects. The C/A@TM developed in this study exhibited promising multifunctional properties as a co-delivery carrier of polypeptide and chemical drug for an effective clinical therapy for cancer.
BACKGROUND:Circadian disruption is a potential cancer risk factor in humans. However, the role of the clock gene, cryptochrome 2 (CRY2), in osteosarcoma (OS) is still not clear. MATERIAL AND METHODS:To evaluate the potential role of CRY2 in HOS osteosarcoma cells, CRY2-silenced cell lines were established. Furthermore, we investigated the effect of CRY2 knockdown on HOS cells by CCK-8, colony formation, migration assay, and flow cytometry, in vitro. RESULTS:CRY2 knockdown promoted HOS OS cell proliferation and migration. We used a cell cycle assay to show that CRY2 knockdown increased the S phase cell population and reduced the G1 phase cell population. Western blot analyses showed that CRY2 knockdown decreased P53 expression and increased expression of c-myc and cyclin D1. Simultaneously, CRY2 knockdown increased the phosphorylation of extracellular signal-regulated kinase (ERK) 1/2, but did not change the phosphorylation of c-Jun N terminal kinase (JNK) and P38. CRY2 knockdown also increased the expression of matrix metalloproteinase (MMP)-2 and β-catenin, and increased OS cell proliferation and migration by inducing cell cycle progression and promoting mitogen-activated protein kinase (MAPK) and Wnt/β-catenin signaling pathways. Although it has previously been unclear whether the expression of CRY2 affects the expression of other clock genes in the clock gene network, our results show that knockdown of CRY2 significantly increased the mRNA expression of CRY1, Period (PER) 1, PER2, BMAL1, and CLOCK. CONCLUSIONS:Our results suggest that CRY2 may be an anti-oncogene in OS, whose functions involve both downstream genes and other circadian genes.
Lilium is the dried bulb of Lilium lancifolium Thunb, Lilium brownii F.E. Brown var.viridulum Baker or Lilium pumilum DC., which belongs to liliaceous plants and has been recorded in China pharmacopoeia 1 .Lilium is a perennial herb and main ornamental plant.In traditional medicine, lilium is well known to treat chronic cough, palpitation, insomnia, etc.Chemical constituents in lilium were begun to study since 1980's.The bulb of lily is rich in nutrient content, such as protein, starch, fat, vitamin B1, B2 and many trace elements.In addition, there are many active matter in the bulb of lily, therefore, lily have extensive pharmacological activity.Besides, steroidal saponins from the bulbs of lilium longiflorum have antitumour-promoter activity 2 and trace colchicine in lily can inhibit the growth and proliferation of cancer cell 3 and have anticancer pharmacological activity.Recently, the researchers gradually pay attention to secondary metabolites in lilium, which can be divided into five groups: glycoside 4-11 , phenolic glycerides, steroidal saponin 12,13 , alkaloid 14 and flavonoids 15 .Compared to the studies on soluble constitutions, the volatile components of lilium have received little attention.Only several volatile components of lilium have been reported using gas chromatography-mass spectrometry [16][17][18][19][20] .GC-MS analysis is a popular and effective method in the qualitative and quantitative analysis of volatile components.However, using only GC-MS, it has not been possible to identify all volatile components.Isolation and identification of volatile components need further work.In addition, the appropriate
Cholesterol metabolism is closely related to the occurrence and development of osteoporosis, but the exact mechanism remains unclear. Niemann-Pick C1-like 1 (NPC1L1) is one of the key cholesterol transporter proteins, however, there are few reports on the functions of NPC1L1 besides regulating cholesterol transport, let alone bone homeostasis. Our preliminary research indicated that NPC1L1 may play a negative regulatory role in osteogenic differentiation. In this study, using in vitro osteogenic differentiation experiment and mouse osteoporosis model, we showed here that NPC1L1 expression was downregulated during osteogenesis, and NPC1L1 knockdown significantly enhanced osteogenic differentiation ability of osteoblasts and delayed progress of osteoporosis. Mechanistically, through RNA sequencing, NPC1L1 was found regulate cholesterol metabolism rather than transportation. It increased 27- Hydroxycholesterol (27-OHC) level through activating 27-hydroxylase (Cyp27a1), resulting in 27-OHC accumulation in osteoblasts and inhibition of osteogenesis. Moreover, C/EBPα was proved mediated NPC1L1 promotes production of 27-OHC by Cyp27a1. These findings reveal that NPC1L1 inhibits osteogenesis and promotes osteoporosis via regulate cholesterol metabolism.
The many circadian clock genes build up a network structure that controls physiological processes, such as the sleep cycle, metabolism, and hormone secretion. Cryptochrome 1 (CRY1), as one of the critical circadian proteins, is closely related to bone formation. However, the regulatory function of CRY1 in osteogenic differentiation remains unclear. In this study, we investigated the role of CRY1 in regulating proliferation and osteoblast differentiation in C3H10 and C2C12 cells after silencing Cry1 using short hairpin RNA interference. In vitro experiments confirmed that the expression level of CRY1 gradually increased during the osteogenic differentiation process, and Cry1 knockdown inhibited the proliferation and differentiation of osteoblastic cells. In addition, Cry1 knockdown inhibited the phosphorylation of AKT kinase (AKT) and extracellular signal-regulated kinase (ERK), which suppressed the phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K)-AKT and mitogen-activated protein kinase (MAPK)-ERK signaling pathways. Taken together, these findings show that CRY1 regulates the proliferation and differentiation of osteoblastic cells in an AKT and ERK-dependent manner.
Abstract Femoral head necrosis is responsible for severe pain and its incidence is increasing. Abnormal adipogenic differentiation and fat cell hypertrophy of bone marrow mesenchymal stem cells increase intramedullary cavity pressure, leading to osteonecrosis. By analyzing gene expression before and after adipogenic differentiation, we found that Microfibril-Associated Protein 5 (MFAP5) is significantly down-regulated in adipogenesis whilst the mechanism of MFAP5 in regulating the differentiation of bone marrow mesenchymal stem cells is unknown. The purpose of this study was to clarify the role of MAFP5 in adipogenesis and therefore provide a theoretical basis for future therapeutic options of osteonecrosis. By knockdown or overexpression of MFAP5 in C3H10 and 3T3-L1 cells, we found that MFAP5 was significantly down-regulated as a key regulator of adipogenic differentiation, and identified the underlying downstream molecular mechanism. MFAP5 directly bound to and inhibited the expression of Staphylococcal Nuclease And Tudor Domain Containing 1, an essential coactivator of PPARγ, exerting an important regulatory role in adipogenesis.
Horizontal cased crossing is the main construction method that is often applied in pipeline projects.In this paper,construction technologies and difficulties in horizontal cased crossing projects were discussed and summarized.And engineering safety,engineering quality control and calculation methods of the parameters were introduced.Besides,the construction method and steps of horizontal cased crossing projects were discussed as well as new construction technology used in the project.
Assuring enough fuel for vehicles is an unavoidable issue.The solution is to develop new fuel.With the strengthening of environmental protection consciousness,the traditional fossil fuel will be replaced by new style fuel,non-fossil fuel.Alcohol fuel,biological diesel oil and fuel battery are all good substitutes.In the paper,present situation and developing tendency of above three kinds of substitutes were analyzed.
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