Differential effects of phenolic extracts from red-fleshed apple peels and flesh induced G1 cell cycle arrest and apoptosis in human breast cancer MDA-MB-231 cells.

Red-fleshedapples are preferredbecause of their high content of phenolics and antioxidants in peel and pulp. Herein, we evaluated the mechanisms of apple peel polyphenolic extracts (APP) and apple flesh polyphenolic extracts (AFP) from the new red-fleshed apple in inhibiting cell proliferation and inducing apoptosis on human breast cancer MDA-MB-231 cells. The antiproliferative activities were determined by the CCK8 assay. The expression of proteins was determined using Western blot. We found that the content of polyphenols and flavonoids in APP was significantly higher than that in AFP, and 14 main phenolic compounds in APP and AFP were quantified using UPLC-MS/MS techniques. Besides, the significant inhibition effects of APP and AFP were achieved through Akt pathway by inducing apoptosis (significantly upregulating reactive oxygen species [ROS] levels, and downregulating expression of pAkt, pBad, Bcl-2, promoting Cytochrome c release, activating Cle-Caspase 9, and inducing expressions of Cle-Caspase 3 and Cle-PARP), and inducing G0/G1 cell cycle arrest (increased expressions of p-p53 and p21 and decreased expressions of PCNA and Cyclin D1). And the inhibition effect of APP was stronger than that of AFP. These results suggest that AFP and APP may be excellent sources of natural chemicals for treating triple-negative breast cancer MDA-MB-231 cells. PRACTICAL APPLICATION: The effects of antiproliferation of phenolic extracts from red-fleshed apple peels and flesh on human breast cancer MDA-MB-231 cells were evaluated. The data may clarify the functional parts of red-fleshed apple and provide some basis for scientific researchers and consumers to recognize and exploit red-fleshed apple.

Involvement of astrocytes activation in orofacial hyperalgesia induced by experimental tooth movement.

OBJECTIVE: The study aimed to investigate the involvement of astrocytes in the medullary dorsal horn (MDH) in the orofacial hyperalgesia induced by experimental tooth movement (ETM) and related mechanism. MATERIALS AND METHODS: Experimental tooth movement was produced with nickel-titanium alloy closed-coil spring fixed between the left maxillary first molar and the left upper incisor. Fluorocitrate was administrated through medullary subarachnoid at 3 days after ETM. Pressure pain threshold (PPT) in masseter cutaneous area was measured. The expression of glial fibrillary acidic protein (GFAP) and c-Fos in MDH was measured using immunofluoroscence staining. The expression of interleukin-1ß (IL-1ß) and phosphorylated N-methyl-D-aspartic acid (NMDA) receptor subunit NR1 (p-NR1) was measured with Western blotting. RESULTS: Experimental tooth movement-induced orofacial hyperalgesia from 1 to 9 days as the PPT was significantly reduced (P < .05). Immunofluoroscence staining showed that the expression of c-Fos in MDH was dramatically upregulated at 1 day and 3 days after ETM, while GFAP expression with both immunofluoroscence staining and Western blotting was significantly enhanced at 3 days and 7 days after ETM. Western blotting analysis indicated that the expression of IL-1ß and p-NR1 in MDH was significantly enhanced at 3 days after ETM. Furthermore, we found that fluorocitrate administration at 3 days after ETM could markedly suppress the expression of c-Fos, GFAP, IL-1ß and p-NR1 and attenuate the reduction of PPT induced by ETM. CONCLUSION: Astrocyte activation in MDH is involved in the mechanical hyperalgesia, and the subsequent upregulated IL-1ß and overexpression of p-NR1 may participate in this process.

A four-year prospective study of self-assembling nano-modified dental implants in patients with type 2 diabetes mellitus.

BACKGROUND/PURPOSE: Dental implantation has become an efficient and important method of replacing lost teeth. However, the success rate of dental-implant treatment in diabetics is higher than patients without diabetes. The aim of this study was to prospectively evaluate long-term marginal bone loss (MBL) and the stability of a self-assembling nano-modified implant in patients with type 2 diabetes mellitus compared with a conventional implant. MATERIALS AND METHODS: Twenty-five patients with type 2 diabetes were recruited for this study. Through a random selection process, one site in each patient received a conventional implant and the other site received a nano-modified implant. The implant stability quotient was measured using resonance frequency analysis (RFA), and MBL was measured using panoramic radiography from uncovering to four-year follow-up. RESULTS: No significant difference in implant stability quotient was found between the two groups (P > 0.05), except for the time at implant insertion (P < 0.05). MBL in the nano-modified implant group exhibited a decreasing change compared with the conventional implant group, between the uncovering and the loading stage (P < 0.05), while there was no significant difference in other stages (P > 0.05). CONCLUSION: There was potentially increased implant stability and diminished MBL around the self-assembling nano-modified implant in the uncovering-loading stage of early osseointegration in patients with type 2 diabetes.

Phytochemical profiles, antioxidant, and antiproliferative activities of red-fleshed apple as affected by in vitro digestion.

The aim of this study was to characterize the phenolic profiles in the extracts and digesta (after in vitro digestion) of different red-fleshed apple fruit parts and to assess the effects of digestion on the in vitro antioxidant capacity and antiproliferative activity. The main polyphenols were identified by UPLC-MS/MS and HPLC. Our results indicate that the digesta had less total phenolics, flavonoids, and anthocyanins, but more free phenolic acids, than the extracts. An analysis of the in vitro antioxidant capacity (including ABTS radical scavenging activity, DPPH radical-scavenging capacity, ferric reducing antioxidant power [FRAP], and cellular antioxidant activity [CAA]) revealed that the digestion decreased the ABTS, DPPH, and FRAP values, but increased the CAA values, relative to the corresponding values for extracts. These results suggest that the digestion improved the effectiveness of the phenolic substances. Moreover, our findings imply that the digestion promoted the antiproliferative activity of red-fleshed apple peels and flesh relative to the extracts. Future in vivo investigations are warranted based on the results of the current study. PRACTICAL APPLICATION: The effects of an in vitro digestion on the phenolic compounds as well as the antioxidative and antiproliferative activities of red-fleshed apple were evaluated. The resulting data may clarify the bioavailability of the polyphenols in red-fleshed apple and enable scientists and consumers to exploit natural polyphenols.

Phytochemical profiles, antioxidant, and antiproliferative activities of four red-fleshed apple varieties in China.

Red-fleshed apples are preferred because of their high content of phenolics and antioxidants. In this study, the phenolic characteristics, antioxidant properties, and antihuman cancer cell properties of the four hybrids of Malus sieversii f. niedzwetzkyana (Ledeb.) M. Roem were analyzed. In addition, the antioxidant and anti-proliferation properties of these apples were measured. Compared to "Fuji" apples, the red-fleshed apples were rich in phenolic and flavonoid chemicals, ranging from 1.5- to 2.6-fold and 1.4- to 2.4-fold, respectively. In all antioxidant methods (DPPH radical-scavenging capacity, ABTS radical scavenging activity, ferric reducing antioxidant power, and cell antioxidant capacity), "A38" obtained the highest antioxidant value, whereas "Fuji" got the lowest antioxidant value. The IC50 values ranged from 33.44 ("A38") to 73.36 mg/mL ("Fuji") for MCF-7 and 20.94 ("A38") to 39.39 mg/mL ("Fuji") for MAD-MB-231. The red-fleshed "A38" and "Meihong" exhibited higher antioxidant and antiproliferative activities in vitro because of the higher levels of phenolics, and the higher potential for development and utilization value. PRACTICAL APPLICATION: The phenolic compounds, antioxidant activity, and antiproliferative activity in vitro of four red-fleshed apple cultivars and one white-fleshed apple cultivar were compared in this study. This information should assist to give a reasonable evaluation for scientists to breed new cultivars with high phenolics and to exploit the natural polyphenol.

XPro1595 ameliorates bone cancer pain in rats via inhibiting p38-mediated glial cell activation and neuroinflammation in the spinal dorsal horn.

Bone cancer pain (BCP) profoundly compromises the life quality of patients with bone metastases. Severe side effects of the drugs which were widely used and effective in the various stages of this condition results in a huge challenge for BCP treatment. Here, we investigated the antinociceptive effects of XPro1595, a soluble tumor necrosis factor (solTNF) inhibitor with considerable immunoregulatory efficacy, on BCP, as well as the underlying mechanisms within the spinal dorsal horn (SDH). Walker 256 mammary gland carcinoma cells were intratibially inoculated to induce BCP. Intrathecal administration of XPro1595 alleviated bone cancer-induced chronic pain in a dose-dependent manner, with an ED50 of 9.69 mg/kg. Bone cancer resulted in the activation of astrocytes and microglia in the SDH through the upregulation of mitogen-activated protein kinase (MAPK) pathways, which was accompanied by an over-expression of pro-inflammatory cytokines, including TNF-α, IL-1ß, and IL-6. XPro1595suppressed bone cancer-evoked glial activation and the consequent neuroinflammation. These inhibitory effects of XPro1595 were, at least partially, mediated by a reduction in the phosphorylation of p38 MAPK in spinal glial cells. In conclusion, inhibition of spinal glia by XPro1595 may have utility in the treatment of bone cancer-induced neuroinflammation, and our results further implicate XPro1595 as a new promising therapeutic agent for BCP.

Investigation of immunogenicity of cryopreserved limbal stem cells.

AIM: To investigate changes in immunogenicity of cryopreserved limbal stem cells. METHODS: Cryopreserved limbal stem cells, fresh primary limbal stem cells and blank controls were inoculated subcutaneously in C57BL-6 mice and the percentage of CD25 cells in limbal explants was determined by flow cytometry at day 21 post inoculation. Morphological studies were performed by light and electron microscopy of limbal explant sections. RESULTS: The number of regional and systemic lymphocytes derived from cryopreserved limbal stem cells was lower than that from fresh primary limbal stem cells. CONCLUSION: Lymphocytes derived from cryopreserved limbal stem cells showed changes in immunogenicity, but the significance is unknown. The cryopreservation and thawing methods await further study.

Sustained release of insulin-like growth factor-1 from poly(lactide-co-glycolide) microspheres improves osseointegration of dental implants in type 2 diabetic rats.

Dental implantation is an effective and predictable treatment modality for replacing missing teeth and repairing maxillofacial defects. However, implants in patients with type 2 diabetes mellitus are likely to have a high failure rate and poor initial osseointegration. In the current study, we established an effective drug delivery system designed to improve osseointegration of dental implants in an animal model of type 2 diabetes. Twenty type 2 diabetic rats were divided into two groups: a group receiving recombinant rat Insulin-like Growth Factor 1 (rrIGF-1) Microsphere Therapy (MST) (10 rats) and a control group (10 rats). The rrIGF-1 was encapsulated into poly(lactide-co-glycolide) (PLGA) microspheres to produce a sustained-release effect around titanium (Ti) dental implants in the rrIGF-1 MST group. Scanning electron microscopy, confocal laser scanning microscopy, and cumulative-release studies were conducted to verify the release effect of the microspheres as well as rrIGF-1 bioactivity. Five rats from each group were sacrificed at weeks 4 and 8 post surgery, and a histological analysis was performed on the rats from both groups. Compared to the control group, rats that received rrIGF-1 by PLGA microsphere treatment were observed to have a higher bone-implant contact percentage around the Ti implants at week 4 or week 8 post surgery (P<0.05). This result clearly indicates that sustained release of rrIGF-1 through encapsulation by PLGA microspheres positively affects osseointegration of dental implants in type 2 diabetic rats.

Type 2 diabetes mellitus impairs bone healing of dental implants in GK rats.

Type 2 diabetes is an increasingly prevalent disease with oral health manifestations. In this study, titanium implants were placed in the femora of 10 type 2 diabetic and 10 age-matched normal rats. We compared the results of bone histomorphometry around the dental implants at 4 and 8 weeks postsurgery.

[Effects of moxibustion at Feishu (BL 13) and Gaohuang (BL 43) on expression of TGF-beta1 in the bleomycin -induced pulmonary fibrosis].

OBJECTIVE: To explore the mechanism of moxibustion arresting the pulmonary fibrosis and provide experimental basis for prevention and treatment of pulmonary fibrosis with acupuncture and moxibustion. METHODS: One hundred and forty SD rats were randomly assigned to 4 groups: a blank group, a model group, a moxibustion group and a prednisone group, 35 rats in each group. The 3 groups expect the blank group were injected with bleomycin via trachea to induce experimental pulmonary fibrosis model, and 7 days after modeling, they were treated with moxibustion at bilateral Feishu (BL 13) and Gaohuang (BL 43), 3 cones each point, once each day, 10 days constituting one therapeutic course with an interval of one day between courses. After 3 courses, all rats were killed and expressions of TGF-beta1mRNA were detected with PCR method. RESULTS: The content of TGF-beta1mRNA in the pulmonary tissue in the moxibustion group and the prednisone group was significantly lower than the model group (P < 0.01), and there was no significant difference between the moxibustion group and the prednisone group (P > 0. 05). CONCLUSION: Both moxibustion at Feishu (BL 13) and Gaohuang (BL 43), and prednisone treatment can significantly suppress the expression of TGF-beta1mRNA in the pulmonary tissue in the rat of bleomycin-induced pulmonary fibrosis.