Silencing PCMT1 enhances the sensitivity of breast cancer cells to paclitaxel through the PI3K/Akt/STMN1 pathway.

This study aimed to investigate whether silencing Protein L-isoaspartate (D-aspartate) O-methyltransferase (PCMT1) expression can enhance the sensitivity of breast cancer cells to paclitaxel and its possible mechanism. Tumor tissues and adjacent histologically normal tissues were collected from patients with breast cancer admitted to our hospital. Human normal breast epithelial cells MCF10A, human breast cancer cells MCF-7, and paclitaxel-resistant breast cancer cells MCF-7/PR were purchased. MCF-7/PR cells were further grouped into negative control (NC) group, si-PCMT1 group (transfected with si-PCMT1), 740Y-P group (treated with 740Y-P, an activator of phosphatidylinositol 3-kinase (PI3K)/ v-Akt Murine Thymoma Viral Oncogene (AKT) signaling pathway), and si-PCMT1 + 740Y-P group (transfected with si-PCMT1 and then treated with 740Y-P). The expression level of PCMT1 in tissues and cells was detected by quantitative real-time polymerase chain reaction (qRT-PCR). Western blot analysis was used to detect the protein expression level of PCMT1 in tissues and cells as well as the protein level of p-PI3K, PI3K, p-Akt, Akt, and Stathmin1 (STMN1) in cells. 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide (MTT) and colony formation assays were used to determine cell viability, scratch assay was used to assess the migration ability of cells, and Transwell assay was used to assess the invasion ability of cells. The expression of PCMT1 was remarkably up-regulated in breast cancer tissues and MCF-7/PR cells. Silencing PCMT1 expression significantly inhibited the proliferation, migration, and invasion of MCF-7/PR cells, and alleviated the resistance of cancer cells to paclitaxel. Additionally, silencing PCMT1 expression also inhibited the activation of PI3K/Akt/STMN1 pathway in MCF-7/PR cells, while activating PI3K/Akt/STMN1 pathway significantly reversed the effect of silencing PCMT1 expression on MCF-7/PR cells. PCMT1 is highly expressed in breast cancer tissues and MCF-7/PR cells, and silencing PCMT1 expression can not only inhibit the development of breast cancer but also enhance paclitaxel sensitivity. Its mechanism of action may be achieved by inhibiting PI3K/Akt/STMN1 signaling.

Formate and CO2 Enable Reductive Carboxylation of Imines: Synthesis of Unnatural α-Amino Acids.

Herein, a photocatalytic umpolung strategy for reductive carboxylation of imines for the synthesis of α-amino acids was disclosed. Carbon dioxide radical anion (CO2•-) generated from formate is the key single electron reductant in the reactions. An unprecedentedly broad substrate scope of imines with excellent reaction yields was obtained with carbon dioxide (CO2) and formate salt as carbon sources.

Nrf2/PHB2 alleviates mitochondrial damage and protects against Staphylococcus aureus-induced acute lung injury.

Staphylococcus aureus (SA) is a major cause of sepsis, leading to acute lung injury (ALI) characterized by inflammation and oxidative stress. However, the role of the Nrf2/PHB2 pathway in SA-induced ALI (SA-ALI) remains unclear. In this study, serum samples were collected from SA-sepsis patients, and a SA-ALI mouse model was established by grouping WT and Nrf2-/- mice after 6 h of intraperitoneal injection. A cell model simulating SA-ALI was developed using lipoteichoic acid (LTA) treatment. The results showed reduced serum Nrf2 levels in SA-sepsis patients, negatively correlated with the severity of ALI. In SA-ALI mice, downregulation of Nrf2 impaired mitochondrial function and exacerbated inflammation-induced ALI. Moreover, PHB2 translocation from mitochondria to the cytoplasm was observed in SA-ALI. The p-Nrf2/total-Nrf2 ratio increased in A549 cells with LTA concentration and treatment duration. Nrf2 overexpression in LTA-treated A549 cells elevated PHB2 content on the inner mitochondrial membrane, preserving genomic integrity, reducing oxidative stress, and inhibiting excessive mitochondrial division. Bioinformatic analysis and dual-luciferase reporter assay confirmed direct binding of Nrf2 to the PHB2 promoter, resulting in increased PHB2 expression. In conclusion, Nrf2 plays a role in alleviating SA-ALI by directly regulating PHB2 transcription and maintaining mitochondrial function in lung cells.

Gastrodia elata and parishin ameliorate aging induced 'leaky gut' in mice: Correlation with gut microbiota.

BACKGROUND: The aging-induced decrease in intestinal barrier function contributes to many age-related diseases. Studies on preventive measures for "leaky gut" may help improve the quality of life of geriatric patients. The potent anti-aging effect of Gastrodia elata and parishin, which is one of its active ingredients, has been reported previously. However, their effects on the gut remain elusive, and the effect of parishin on mammals has not been studied. METHODS: We used quantitative RT-PCR, western blotting, immunohistochemical analysis, and 16S rRNA sequencing to investigate the effect of G. elata and parishin on the intestinal barrier function of D-Gal-induced aging mice. RESULTS: G. elata and parishin prevented the decrease in tight junction protein (TJP) expression and morphological changes, modulated the composition of fecal microbiota to a healthier state, and reversed the translocation of microbial toxins and systemic inflammation. The correlation analyses showed that TJP expression and systemic inflammation were significantly positively or negatively correlated with the composition of fecal microbiota after G. elata and parishin administration. Additionally, TJP expression was also correlated with systemic inflammation. Moreover, G. elata and parishin administration reversed the decreased or increased expression of aging-related biomarkers, such as FOXO3a, SIRT1, CASPASE3 and P21, in the gut. CONCLUSIONS: These results suggested that G. elata and parishin could prevent gut aging and ameliorate the "leaky gut" of aged mice and that the underlying mechanism is related to the mutual correlations among barrier function, fecal microbiota composition, and inflammation.

A Clinical Application Study of Nanostructured Substrates for the Detection of Circulating Tumor Cells.

BACKGROUND: Circulating tumor cell (CTC) isolation methods based on nanostructured substrates can be used to isolate tumor cells from peripheral blood. This study aimed to validate the clinical application of our method and determine the appropriate diagnostic critical value. METHODS: AFM was used to detect the surface roughness of nanostructured substrates. Cell lines and blood samples were used to verify CTC isolation methods. The ROC curve and AUC were used to evaluate the diagnostic value of CTC numbers. RESULTS: First, AFM, cell binding yields, and tumor cell detection rate from blood showed that NS has a potential for cell adsorption. Then, the CTC detection method was verified by using cell lines and blood samples. The number of CTCs in patients with cancers or metastases were significantly greater than those of patients without cancers. Then, the ROC curves and AUC showed that this method had a medium diagnostic value. CONCLUSIONS: Isolating CTCs based on nanostructured substrates was appropriate for the clinical diagnosis of tumors, and samples with more than 1.5 CTCs/1 mL blood could be identified as CTC-positive.

Cryptoporus volvatus polysaccharides attenuate LPS-induced expression of pro-inflammatory factors via the TLR2 signaling pathway in human alveolar epithelial cells.

CONTEXT: Cryptoporus volvatus (Peck) Hubb grows wild in China, and its fruiting bodies have been used traditionally to treat asthma and bronchitis. OBJECTIVES: This study evaluates the anti-inflammatory effect of Cryptoporus polysaccharides (CP) extracted from fruiting bodies of C. volvatus on lipopolysaccharide (LPS)-induced pro-inflammatory factors and the signaling pathways involved in human alveolar epithelial cells. MATERIALS AND METHODS: To evaluate the effects of CP on LPS-induced pro-inflammatory factors, A549 cells were pre-incubated with CP 1, 10, and 100 µg/ml for 1 h and then stimulated with LPS 10 µg/ml for 24 h. The expression of pro-inflammatory factors monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), Toll-like receptor 2 (TLR2), and phosphorylation of ERK1/2, p38, and NF-κB p65 were measured by q-PCR, ELISA, and western blotting. RESULTS: CP decreased LPS-induced mRNA expression of MCP-1, TNF-α, and IL-1ß (IC50 = 83.3, 85.2, and 91.6 µg/ml, respectively) and their correspondent protein expression (IC50 = 88.6, 76.4, and 81.6 µg/ml, respectively). Investigation of potential mechanisms indicated that CP 100 µg/ml reduced LPS-induced expression of TLR2 mRNA (66.9%, p < 0.01) and protein (63.2%, p < 0.01) that was a result of the decreased pro-inflammatory factors. LPS induction increased the expression of TLR2 and the phosphorylation of p38 and ERK1/2, NF-kB p65 concomitantly. CP 100 µg/ml inhibited the LPS-induced phosphorylation of the signaling proteins (p < 0.05). CONCLUSIONS: This suggests that CP pretreatment down-regulates LPS-mediated inflammation in lung epithelial cells. This study further confirmed that CP is a potential anti-inflammatory drug for the treatment of airway inflammatory diseases.

Apple Polyphenols Decrease Atherosclerosis and Hepatic Steatosis in ApoE-/- Mice through the ROS/MAPK/NF-κB Pathway.

In this study, we examined the effects of apple polyphenols (APs) on hyperlipidemia, atherosclerosis, hepatic steatosis and endothelial function and investigated the potential mechanisms. ApoE(-/-) mice were fed a western-type diet and orally treated with APs (100 mg/kg) or atorvastatin (10 mg/kg) for 12 weeks. Hyperlipidemia and atherosclerosis in the aortic sinuses and, and hepatic lipidosis were measured. The treatment with APs or atorvastatin induced a remarkable reduction in the atherosclerotic lesions and hepatic steatosis and decreased the levels of low-density lipoprotein, triglyceride, CCL-2 and VCAM-1 levels in the plasma. Conversely, the APs significantly increased the plasma levels of high-density lipoprotein (HDL) cholesterol and markedly up-regulated the glutathione peroxidase (GPx), catalase (CAT) and superoxide dismutase (SOD) levels in liver tissues. Moreover, the APs treatment modulated lipid metabolism by up-regulating the transcription of associated hepatic genes including PPARα, while down-regulating the transcription of SCAP and its downstream genes associated with lipid synthesis in the liver. Histological assessment showed that the APs treatment also reduced the macrophage infiltration in the aortic root plaque and the inflammatory cells infiltrations to the liver tissues. Moreover, we confirmed that the APs treatment greatly reduced the ox-LDL-induced endothelial dysfunction and monocyte adhesion to rat aortic endothelial cells (RAECs). Mechanistically, the APs treatment suppressed the ROS/MAPK/NF-κB signaling pathway, and consequently, reduced CCL-2, ICAM-1 and VCAM-1 expression. Our results suggest that the APs are a beneficial nutritional supplement for the attenuation of atherosclerosis.

Pre-expanded thoracodorsal artery perforator-based flaps for repair of severe scarring in cervicofacial regions.

BACKGROUND: Reconstruction of cervicofacial scarring continues to present challenges for surgical treatment. Here we present our clinical experience in repairing cervicofacial scarring using pre-expanded thoracodorsal artery perforator flaps. METHODS: From January 2007 to December 2012, 15 patients were treated for severe cervicofacial scarring. In the first surgical stage, expanders were implanted subcutaneously in the zone nourished by thoracodorsal artery perforators. The expansion generally took 3 to 6 months. In the second surgical stage, the cervicofacial cicatricial contracture was released and the secondary defect was covered with local flaps. The remaining wound was covered by the free thoracodorsal artery perforator expanded flap, which was anastomosed to the facial vascular bundle. The donor site was closed directly in all the patients. RESULTS: The postoperative follow-up time ranged from 1 to 5 years. The deformities were corrected, all flaps survived completely and none were bulky. The maximum length of the flaps was 32 cm (mean, 22.4 ± 4.2 cm), and the maximum width was 17 cm (mean, 14.4 ± 2.2 cm). All patients exhibited recovery of neck movement, and there was no recurrence of neck contracture. CONCLUSION: The pre-expanded thoracodorsal artery perforator flap is an ideal method for reconstruction of severe cervicofacial cicatricial contracture.

[Repair of tissue defects with free composite anterolateral femoral fascia lata perforator tissue flaps].

OBJECTIVE: To observe the clinical effects of repair of complicated tissue defects of several body parts with composite anterolateral femoral fascia lata perforator tissue flaps (fascial flap or fascial skin flap) with the aid of micro-surgery. METHODS: From February 2008 to August 2012, complicated tissue defects in 12 patients were repaired with composite anterolateral femoral fascia lata perforator tissue flaps. Two of the 12 patients suffered from a defect of scalp, skull, and dura mater as a result of resection of a malignant tumor of the scalp; 3 patients showed a defect of skin and tendo calcaneus in the heel and lower leg; 2 patients showed a defect of skin and extensor tendon in the dorsum of hands; the other 5 patients suffered from defects of skin and extensor tendon in the foot and ankle combined with exposure of bone or internal buttress plate. The size of tissue flaps ranged from 12 cm ×6 cm to 19 cm ×18 cm. The donor sites were closed by immediate suturing or skin grafting. RESULTS: All 12 tissue flaps survived. Patients were followed up for 2 to 36 months. The flaps were shown to have good appearance, texture and function. Two patients with the defect of the scalp, skull and dura mater after a resection of the malignant tumor of the scalp did not have recurrence or herniation of brain tissue. The foot-raising function in 3 patients with the defect of skin and tendo calcaneus in the heel and lower leg was recovered, and according to Arner-Lindholm criteria the result was excellent in 2 cases and good in 1 case. The extension function of fingers of 2 patients with defects of skin and extensor tendon in the dorsum of hands was good according to the evaluation criteria of Chinese Medical Association Society of Hand Surgery for tendon repair of hand. The extension function of toes of 5 patients with defects of skin and extensor tendon in the foot and ankle combined with exposure of bone or internal buttress plate was recovered and improved. CONCLUSIONS: Transplantation of composite anterolateral femoral fascia lata perforator tissue flaps with the aid of micro-surgery is an effective method in repairing the tissue defects of skull, dura mater, and the extensor tendon of hands or feet, with restoration of the extension function.

Effects of ORP150 on appearance and function of pancreatic beta cells following acute necrotizing pancreatitis.

Pancreatic beta cells produce and release insulin, which decreases the blood glucose level. Endoplasmic reticulum stress caused pancreatic beta cell dysfunction and death in acute necrotizing pancreatitis (ANP). The 150kD oxygen-regulated protein (ORP150) took part in the process of endoplasmic reticulum stress. This study investigated the effect of ORP150 on appearance and function of pancreatic beta cells in ANP. Acute necrotizing pancreatitis relied on retrograde infusion of 5% sodium taurocholate into the bile-pancreatic duct. The severity of ANP was estimated by serum amylase, secretory phospholipase A(2,) and pancreatic histopathology. The changes in appearance and function of pancreatic beta cells were detected by light and electron microscopy and the levels of serum glucose, insulin, and C-peptide. ORP150 expression was studied using western blot and immunohistochemisty assay. The expression of ORP150 mainly appeared on pancreatic beta cells and decreased gradually during the pathogenesis of ANP. The results of light and electron microscopy indicated pancreatic beta cell dysfunction and death, concomitant with elevation of serum glucose, insulin, and C-peptide in ANP. These results imply a probable role of ORP150 in the changes in appearance and function of pancreatic beta cells following acute necrotizing pancreatitis, through the pathway of endoplasmic reticulum stress.

[Fabricated expanded thoracodorsal artery perforator flap to repair cervical scar in children].

OBJECTIVE: To investigate ideal methods to repair cervical cicatricial contracture in children. METHODS: The expanders were implanted subcutaneously around the cervical scar and above the latissimus. After expansion was completed, the cervical cicatricial contracture was released and the wound was covered with local expanded flaps and free expanded prefabricated thoracodorsal artery perforator flap, leaving no injury to thoracodorsal nerves and latissimus. The wound at the donor site was closed directly. RESULTS: From July 2007 to October 2009, 10 patients were treated. All the flaps survived completely. All the wounds were repaired totally and the deformities were corrected completely. The patients were followed up for 3-30 months. When the patients grew up, the flaps enlarged simultaneously. The flaps were not bulky and had a good color match. The scar at the donor site was inconspicuous with no functional morbidity. CONCLUSION: The fabricated expanded thoracodorsal artery perforator flaps is an ideal method for severe cervical cicatricial contracture in children.