Integrated Analysis of ECT2 and COL17A1 as Potential Biomarkers for Pancreatic Cancer.

Background: Pancreatic cancer (PC) is a malignant tumor of the digestive tract. It presents with atypical clinical symptoms and lacks specific diagnostic indicators. This study is aimed at exploring the potential biomarkers of PC. Methods: TCGA database pancreatic cancer dataset was normalized and used to identify differentially expressed genes (DEGs). Survival, independent prognostic, and clinical correlation analyses were performed on DEGs to screen for key genes. DNA methylation, mutation, and copy number variation (CNV) analyses were used to analyze genetic variants in key genes. GSEA was performed to explore the functional enrichment of the key genes. Based on the expression of key genes, construction of a competing endogenous RNA (ceRNA) network, analysis of the tumor microenvironment (TME), and prediction of chemotherapeutic drug sensitivity were performed. Furthermore, the GEO database was used to validate the reliability of key genes. Results: Two key genes (ECT2 and COL17A1) were identified, which were highly expressed in PC. The mRNA expression of ECT2 and COL17A1 was associated with DNA methylation and CNV. The cell cycle, proteasome, and pathways in cancer were enriched in the high-COL17A1 and ECT2 groups. The TME results showed that immune scores were decreased in the high-ECT2 group. CeRNA network results showed that there were eleven miRNAs were involved in the regulation of ECT2 and COL17A1. Moreover, pRRophetic analysis showed that 20 chemotherapeutic drugs were associated with ECT2 and COL17A1 expression. Conclusions: Collectively, ECT2 and COL17A1 may be potential biomarkers for PC, providing a new direction for clinical diagnosis and treatment.

Allicin enhances the radiosensitivity of colorectal cancer cells via inhibition of NF-κB signaling pathway.

Radioresistance is an important factor affecting the radiotherapy effect of colorectal cancer (CRC). Allicin is a versatile sulfur-containing organic compound extracted from garlic (Allium sativum L.), which has many pharmacological effects. However, the effect of allicin on the sensitivity of CRC radiotherapy has not been confirmed. The present study is to observe the radiosensitivity effects of allicin and to explore its mechanism in CRC radiotherapy. The proliferation inhibition effects of allicin combined with X-ray radiotherapy in HCT116 cells were measured by growth curve of cell and colony formation assays. The cell apoptosis was detected by Hoechst 33258 nucleus staining assay. The migration ability of cells was detected by Transwell chamber migration assay. The animal model of CRC was established in BALB/c mice via transplantation of CT26 cell, and the radiosensitization effect of allicin on CRC was detected in vivo. The mRNA expressions of NF-κB, IKKß, and IκBα were analyzed by reverse transcription-polymerase chain reaction (RT-PCR). The protein expressions of NF-κB, p-NF-κB, IKKß, p-IKKß, IκBα, and p-IκBα were detected by western blotting. Our results showed that allicin improves the sensitivity of X-ray radiotherapy in CRC, and its mechanism may be associated with inhibition of NF-κB signaling pathway. These findings suggest that allicin may be used as a potential sensitizer for tumor radiotherapy in the clinic.

Composite scaffolds loaded with bone mesenchymal stem cells promote the repair of radial bone defects in rabbit model.

This study aimed to investigate the efficacy of three-dimensional scaffolds of silk fibroin/chitosan/nano-hydroxyapatite (SF/CS/nHA) and bone marrow derived mesenchymal stem cells (BMSCs) on the repair of long segmental bone defects in rabbits. BMSCs were cultured with SF/CS/nHA in vitro, and cell proliferation, alkaline phosphatase activity and Ca2+ content were examined. A 15mm segmental defect in the radius was generated in 12 New Zealand White rabbits, which were divided randomly into three groups (n=4): experimental group with SF/CS/nHA scaffold of induced BMSCs; control group with SF/CS/nHA scaffold; and blank group without any materials. Postoperatively at 12 weeks, osteogenesis effect and the degradation and absorption of SF/CS/nHA were evaluated by X-ray, hematoxylin eosin staining, and scanning electron microscopy. In vitro, SF/CS/nHA scaffolds exhibited good biocompatibility and no toxicity. SF/CS/nHA promoted adhesion, growth, and calcium nodule formation of BMSCs compared to control (P<0.05). In vivo, we observed gradual new bone formation and bone defect gradually recovered at 12 weeks in experimental and control group, but more new bone was formed in experimental group (P<0.05). In blank group, limited bone formation was observed and bone defect was obvious. In conclusion, SF/CS/nHA scaffolds loaded with BMSCs achieve high efficacy to repair segmental defect in the radius.

Application of silk fibroin/chitosan/nano-hydroxyapatite composite scaffold in the repair of rabbit radial bone defect.

Silk fibroin (SF), chitosan (CS) and nano-hydroxyapatite (nHA) possess excellent biocompatibility, thus, these were used to construct a SF/CS/nHA composite scaffold. Previously published results identified that this material exhibited satisfactory physical and chemical properties, and therefore qualified as a repair material in bone tissue engineering. The aim of the present study was to investigate the capacity and mechanism of this composite scaffold in repairing bone defects. In total, 45 New Zealand white rabbits were used to model defect in the right radial bone. A radial bone defect was induced, and rabbits were divided into the following treatment groups (n=15 in each): Group A, in which the SF/CS/nHA scaffold was implanted; group B, in which the SF/CS scaffold was implanted; and group C, in which rabbits did not receive subsequent treatment. X-ray scanning, specimen observation and histopathological examination were implemented at 1, 2, 3 and 4 months after modeling, in order to evaluate the osteogenic capacity and mechanism. At 1 month after modeling, the bone density shadow in the X-ray scan was darker in group A as compared with that in group B. Observation of the pathological specimens indicated that normal bone tissues partially replaced the scaffold. At 2 months, the bone density shadow of group A was similar to normal bone tissues, and normal tissue began to replace the scaffold. At 3-4 months after modeling, the X-ray scan and histopathological observation indicated that the normal bone tissues completely replaced the scaffold in group A, with an unobstructed marrow cavity. However, the bone mass of group B was lower in comparison with that of group A. The bone defect induced in group C was filled with fibrous connective tissues. Therefore, it was concluded that the SF/CS/nHA composite scaffold may be a promising material for bone tissue engineering.

Preparation of a biphase composite scaffold and its application in tissue engineering for femoral osteochondral defects in rabbits.

PURPOSE: Three-dimensional bioactive scaffolds are useful tools for stem cell implant in tissue-engineering. For chondral and subchondral repair, the chondroinductive and osteoinductive property of a scaffold is a major challenge. The scaffolds that aim to osteogenic differentiation have been well studied. However, cartilage cells can hardly be induced for osteogenesis, and monophase scaffolds cannot ideally repair both cartilage and subchondral defects at the same time. METHODS: We developed a novel biphase composite scaffold and observe its application osteochondral defects. We combined the advantages of silk-fibroin/chitosan (SF/CS) scaffold in chondrogenic differentiation and the silk-fibroin/chitosan/nano-hydroxyapatite (SF/CS/nHA) scaffold in osteogenic differentiation and bone regeneration, and synthesized a SF/CS-SF/CS/nHA scaffold, which contained both the chondrocytic phase (SF/CS) and the osteoblastic phase (SF/CS/nHA). RESULTS: The biphase scaffold exhibited a porosity ratio around 90% and a water absorption ratio about 822%. A similar degradation property to traditional monophase scaffolds was observed. Bone mesenchymal stem cells (BMSCs) showed a good proliferation on this scaffold. Expression of two types of collagen was inducable for BMSCs on the scaffold. Neoformative extracellular matrix integrated with the scaffold was observed by the scanning electron microscope. When implanted in the lesion site in the rabbit femur with cartilage injury, mixing and filling function were exerted by the cell-scaffold constructs (CSCs). Micro-CT scanning revealed both chondral and subchondral layers were repaired. Moreover, type I and II collagens were both expressed in the implanted CSCs. CONCLUSIONS: Chondral and subchondral repair can be achieved using the biphase scaffold implant that permits both chondrogenesis and osteogenesis from BMSCs. This approach has the potential to be clinically used for tissue engineering implantation.

Tanshinone IIA induces intrinsic apoptosis in osteosarcoma cells both in vivo and in vitro associated with mitochondrial dysfunction.

Tanshinone IIA (Tan IIA), a phytochemical derived from the roots of Salvia miltiorrhiza, has been shown to inhibit growth and induce apoptosis in various cancer cells. The association of its inhibitory effect on the primary malignant bone tumor, osteosarcoma, with mitochondrial dysfunction remains unclear. This study aimed to investigate the anti-proliferative effects of Tan IIA on human osteosarcoma 143B cells both in vitro and in vivo. Administration of Tan IIA to NOD-SCID mice implanted with 143B cells led to significant inhibition of tumor development. The inhibition of proliferation, migration, and invasion was observed in 143B cells treated with Tan IIA. The tumor proliferation markers, Ki67 and PCNA, were suppressed and apoptosis by TUNEL assay was activated respectively. Apoptosis in the Tan IIA-treated 143B cells and xerograft mice was associated with the activation of caspase cascade via the modulation of Bcl-2 family. The CD31 was inhibited in Tan IIA-treated xenografts to indicate anti-neovasculization. Tan IIA administration resulted in a significant decrease in the mitochondrial fusion proteins, Mfn1/2 and Opa1, as well as an increase in the fission protein Drp1. We concluded that mitochondrial dysfunction associated with dynamic change was involved in apoptosis and anti-angiogenesis elicited by Tan IIA.

Phyllanthus urinaria's Inhibition of Human Osteosarcoma Xenografts Growth in Mice is Associated with Modulation of Mitochondrial Fission/Fusion Machinery.

Osteosarcoma is an aggressive bone cancer arising from primitive transformed cells of mesenchymal origin to form malignant osteoid. Phyllanthus urinaria [Formula: see text]P. urinaria[Formula: see text] is a widely used folk medicine in cancer treatment, however the mechanism of P. urinaria inhibited human osteosarcoma is unclear. The present study was aimed at investigating the antitumoral effects of an aqueous P. urinaria on human osteosarcoma in vivo and the related underlying mechanisms, mainly focusing on mitochondrial dynamic dysfunction. Our results showed that oral administration of P. urinaria to mice led to significant inhibition of tumor development without substantial changes to body weight or major organs. Histological examinations with H&E, Giemsa, and Masson trichrome stains confirmed inhibition of tumor growth by the P. urinaria treatment. Immunohistochemical staining of proliferation markers antigen KI-67 (Ki67) and proliferating cell nuclear antigen (PCNA), as well as a terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay demonstrated a decrease of tumor proliferation and an increase of apoptosis, which was associated with the modulation of B-cell lymphoma 2 (Bcl-2) family activating the caspase cascade in the P. urinaria-treated mice. The neovascularization marker cluster of differentiation 31 (CD31) was inhibited in P. urinaria-treated xenografts, implicating the potential anti-angiogenic effect of P. urinaria. P. urinaria treatment resulted in a significant decrease in the mitochondrial fusion proteins, including mitofusin 1/2 (Mfn1/2) and optic atrophy type 1 (Opa1), as well as an increase in the fission protein dynamin-related protein 1 (Drp1). The results of this study suggest mitochondrial dysfunction is associated with dynamic change that is involved in the apoptosis and anti-angiogenesis elicited by P. urinaria.

Association of toll-like receptor 9 gene polymorphism in Chinese patients with systemic lupus erythematosus in Taiwan.

The purpose of this study was to determine whether toll-like receptor 9 (TLR9) gene polymorphisms were markers of susceptibility to or severity of systemic lupus erythematosus (SLE) in Taiwanese patients. The study included 211 healthy individuals and 167 Chinese patients with SLE. Polymorphisms of TLR9 [rs2066807 and rs187084 (-1486 T/C)] were typed from genomic DNA. The genotypes, allelic frequencies, and carriage rates were compared between patients with SLE and control subjects. The relationship between allelic frequencies and clinical manifestations of 167 patients with SLE was evaluated. There was no statistically significant difference in TLR9 (rs2066807) gene polymorphism, allelic frequency, and carriage rate between the SLE and control groups. However, for the genotype of TLR9 -1486 T/C (rs187084) polymorphism, there was a statistically significant difference between the SLE and the control groups (P < 0.001, χ(2) = 15.9). Moreover, there was a significant association between the two groups in allelic frequency and carriage rate of the T allele (P < 0.001, χ(2) = 18.5 and P < 0.01, χ(2) = 8.06, respectively). We did not detect any association between the TLR9 genotype and the clinical or laboratory profiles in patients with SLE. The results suggest that the TLR9 -1486 T/C (rs187084), but not TLR9 (rs2066807), polymorphism is related to SLE in Taiwanese patients.

MBD4 gene is associated with rheumatoid arthritis in Chinese patients in Taiwan.

This study examines whether or not MBD4 polymorphism is a marker for rheumatoid arthritis (RA) susceptibility or severity for Chinese patients in Taiwan. This study included 193 patients with RA, while 190 unrelated healthy individuals living in Central Taiwan served as controls. The relationship between MBD4 polymorphism and clinical manifestations of RA was evaluated. For the genotype and allelic frequency of MBD4-1057 polymorphism, there were no statistically significant differences between patients with RA and controls. There were significant differences in the distribution of MBD4-8666 polymorphism frequencies between patients with RA and controls [P = 0.013; P (corrected) (Pc) = 0.039]. There were also significant relationships in the distribution of MBD4-9229 polymorphism genotype between patients with RA and controls (P = 0.007; Pc = 0.021). However, we did not detect any associations for MBD4-1057, MBD4-8666 or MBD4-9229 with rheumatoid factor presence, extra-articular involvement or bone erosion in patients with RA. Results suggest that MBD4-8666 and MBD4-9229, but not MBD4-1057, gene polymorphisms are related to RA in Chinese patients in Taiwan.

p53 codon 72 proline/arginine polymorphism and autoimmune thyroid diseases.

p53 protein participates in the processes of apoptosis, which is involved in a number of immunological reactions. In order to test whether the p53 gene could be used as a genetic marker for the prediction of the development of autoimmune thyroid diseases (AITD), we screened, by using polymerase chain reaction (PCR) analysis, for the C (CCC)/G (CGC) polymorphism at the p53 codon 72 (Pro 72/Arg 72) to determine the genotypes of 107 Hashimoto's thyroiditis (HT) and 90 Graves' disease (GD) patients, and 105 normal controls. The data demonstrated that, for the genotype analysis, HT patients featured an enhanced numerical ratio for the Arg/Arg homozygous genotype (33.7%) and a diminished ratio for the Arg/Pro heterozygous genotype (41.1%) at the p53 codon 72 than was the case for normal controls (Arg/Arg: 17.1% and Arg/Pro: 61.9%; P=0.005). The odds ratio for the risk of the Arg/Arg genotype's appearance, compared with that of the Arg/Pro and Pro/Pro genotypes combined, for the HT patient group was 2.450 (95% confidence interval: 1.274-4.716). With respect to allelic analysis, we did not observe significant difference in the frequency of appearance of the Arg allelic variant and the Pro allelic variant for the p53 codon 72 when comparing the HT patient group with the control group (P=0.208). On the other hand, GD patients presented no significant difference in distribution for both genotype and allelic frequencies (P=0.344 and 0.245, respectively) when compared with normal controls. In conclusion, HT patients feature a greater ratio of arginine homozygosity at p53 codon 72 than in the case for normal subjects. The p53 codon 72 proline/arginine polymorphism may be a genetic marker to predict the increased susceptibility of development of HT.

Photophysical and electrochemical properties of new ortho-metalated complexes of rhodium(III) containing 2,2'-dipyridylketone and 2,2'-dipyridylamine. An experimental and theoretical study.

Two new ortho-metalated rhodium(III) complexes of the formula [Rh(ppy)(2)(L)](+), ppy = 2-phenylpyridine and L = 2,2'-dipyridylketone (dpk) (), 2,2'-dipyridylamine (HDPA) () have been synthesized and subjected to X-ray diffraction crystal structural, photophysical and electrochemical studies. Density functional theory calculations have also been performed to get rationalizations of the optical orbitals and redox orbitals concerning photophysical and electrochemical data. Complex exhibits the triplet ligand-to-ligand charge transfer ((3)LLCT) [pi(ppy)-pi*(dpk)] phosphorescence at 77K (520 nm) and at room temperature (555 nm), while complex shows triplet ligand centred ((3)LC) [pi-pi*(ppy)] phosphorescence only at 77K (460 nm). Both complexes and have similar irreversible oxidation potentials (+1.19 V for and +1.15 V for vs. Fc/Fc(+)). These two complexes show different characteristics in the reduction process: a reversible process occurs for at -1.31 V, while an irreversible process is observed for 2 at -1.85 V.