Protective effects of sodium ferulate on flap transplantation via anti-inflammatory modulation and oxidative stress inhibition.

Ischemia-reperfusion injury (IRI) has brought attention to flap failure in reconstructive surgery. To improve the prognosis of skin transplantation, we performed experimental IRI by surgical obstruction of blood flow and used sodium ferulate (SF) to prevent IRI in rats. After SF treatment, the morphological and histological changes of the skin flaps were observed by H&E and Masson's trichrome staining. We also detected the expression levels of COX-1, HO-1, and Ki67 by immunohistochemical and western blot analysis. Moreover, enzyme-linked immunosorbent assay was used to identify the content of tumor necrosis factor (TNF)-α, myeloperoxidase (MPO), malondialdehyde (MDA), and nitric oxide (NO) in peripheral blood and skin tissue. Compared with the model group, SF treatment significantly improved the recovered flap area (%) and promoted collagen synthesis. Cyclooxygenase-2 (COX-2) expression was significantly inhibited by heme oxygenase-1 (HO-1) induction after SF treatment. Furthermore, SF significantly inhibited the levels of TNF-α in peripheral blood, MPO and MDA in the skin tissue, and the increased synthesis of NO. Our results showed the protective effects of SF on IRI after flap transplantation and we believe that the protective effects of SF was closely related to the alleviation of the inflammatory response and the inhibition of the oxidative stress injury.

Protective effects of sodium ferulate on flap transplantation via anti-inflammatory modulation and oxidative stress inhibition

Ischemia-reperfusion injury (IRI) has brought attention to flap failure in reconstructive surgery. To improve the prognosis of skin transplantation, we performed experimental IRI by surgical obstruction of blood flow and used sodium ferulate (SF) to prevent IRI in rats. After SF treatment, the morphological and histological changes of the skin flaps were observed by H&E and Masson's trichrome staining. We also detected the expression levels of COX-1, HO-1, and Ki67 by immunohistochemical and western blot analysis. Moreover, enzyme-linked immunosorbent assay was used to identify the content of tumor necrosis factor (TNF)-α, myeloperoxidase (MPO), malondialdehyde (MDA), and nitric oxide (NO) in peripheral blood and skin tissue. Compared with the model group, SF treatment significantly improved the recovered flap area (%) and promoted collagen synthesis. Cyclooxygenase-2 (COX-2) expression was significantly inhibited by heme oxygenase-1 (HO-1) induction after SF treatment. Furthermore, SF significantly inhibited the levels of TNF-α in peripheral blood, MPO and MDA in the skin tissue, and the increased synthesis of NO. Our results showed the protective effects of SF on IRI after flap transplantation and we believe that the protective effects of SF was closely related to the alleviation of the inflammatory response and the inhibition of the oxidative stress injury.

Deep RNA sequencing elucidates microRNA-regulated molecular pathways in ischemic cardiomyopathy and nonischemic cardiomyopathy.

Deregulation of cardiac miRNA gene-regulatory networks is a feature of different heart diseases, including ischemic (ICM) and nonischemic (NICM) cardiomyopathy. Here, based on the paired miRNA and mRNA expression profiles in ICM and NICM, we identified the differentially expressed miRNAs and mRNAs and the expression signatures distinguishing ICM/NICM from control samples. Furthermore, we constructed a functional miRNA network for each disease. Analysis of the topological features of these networks revealed that the Wnt signaling pathway and cell cycle (de)regulation play critical roles in the development of ICM and NICM. In addition, comparison of the miRNA and mRNA functional profiles revealed that their expression patterns in ICM and NICM differ. These findings revealed hundreds of novel heart-failure-related miRNAs with important regulatory functions. In summary, RNA-seq-based transcriptome profiling in the failing human heart revealed a complex transcriptional regulation associated with the disease. The newly uncovered importance of miRNAs in disease pathogenesis highlights their value as potential diagnostic and therapeutic targets.

A new method for estimating the number of non-differentially expressed genes.

Control of the false discovery rate is a statistical method that is widely used when identifying differentially expressed genes in high-throughput sequencing assays. It is often calculated using an adaptive linear step-up procedure in which the number of non-differentially expressed genes should be estimated accurately. In this paper, we discuss the estimation of this parameter and point out defects in the original estimation method. We also propose a new estimation method and provide the error estimation. We compared the estimation results from the two methods in a simulation study that produced a mean, standard deviation, range, and root mean square error. The results revealed that there was little difference in the mean between the two methods, but the standard deviation, range, and root mean square error obtained using the new method were much smaller than those produced by the original method, which indicates that the new method is more accurate and robust. Furthermore, we used real microarray data to verify the conclusion. Finally we provide a suggestion when analyzing differentially expressed genes using statistical methods.

Platelet parameters and expression of platelet membrane glycoprotein in childhood acute lymphoblastic leukemia.

Platelet activation and functional changes were investigated in acute lymphoblastic leukemia (ALL) to provide the basis for early diagnosis and evaluation of curative effect. Platelet parameters, immature platelet fraction (IPF%), immature platelet count (IPC), P-selectin (CD62p) (PAC-1) expression were detected in ALL, ALL-complete remission-induced (CR1), and normal groups with an automatic blood cell analyzer and flow cytometer. CD62p and PAC-1 were higher in the ALL group before adenosine-5-diphosphate (ADP) activation than in the normal group (P < 0.05); PAC-1 expression was higher and lower in the ALL-CR1 group than in normal and ALL groups (P < 0.05), respectively. CD62p and PAC-1 expression was lower in the ALL group than in the normal group after ADP activation (P < 0.05); PAC-1 expression was lower and higher in the ALL-CR1 group than in normal and ALL groups, respectively (P < 0.05). Platelet count (PLT), mean platelet volume (MPV), platelet hematocrit (PCT), and platelet distribution width (PDW) were lower in the ALL group than in the ALL-CR1 group (P < 0.05). PLT, MPV, and PCT did not differ between the ALL-CR1 group and the normal group (P > 0.05). PDW did not differ statistically among all groups (P > 0.05). IPF% and IPC values were higher and lower in the ALL group than in normal and ALL-CR1 groups (P < 0.05), respectively. These did not differ significantly between the normal group and the ALL-CR1 group (P > 0.05). Therefore, ALL patients demonstrate platelet activation and platelet dysfunction; platelet parameters and membrane glycoprotein expression can be used to evaluate the effect of ALL.

Effect of CYP1A1 and GSTM1 genetic polymorphisms on bone tumor susceptibility.

Tumor gene polymorphisms are often associated with individual susceptibility to genetic diseases. Cytochrome P4501A1 (CYP1A1) and glutathione S-transferase mu 1 (GSTM1) gene polymorphisms are closely related to the susceptibility of the body to chemical carcinogens in the environment. Therefore, we explored the relationship between CYP1A1 and GSTM1 gene polymorphisms and susceptibility to bone tumors. Multiplex-polymerase chain reaction (PCR), allelic-specific PCR, and PCR-restriction fragment length polymorphism techniques were used to analyze CYP1A1 and GSTM1 gene polymorphisms in 52 bone tumor patients and 100 healthy subjects. The allelic variation frequency of the CYP1A1 gene at exon 7 (Ile 462 Val) in bone tumor patients was 0.462, which was significantly higher than that in the normal controls (0.223). The frequency of the absence of the GSTM1 homozygous genotype in the patients (0.65) was also markedly higher than that in the control group (0.41). Subjects with CYP1A1 Val/Val homozygous mutations and absence of the GSTM1 homozygous genotype were at markedly increased risk of developing bone tumors [ORs 4.15 (95%CI: 1.268-13.30) and 2.35 (95%CI: 1.15-4.85), respectively]. The OR for the combined effect of the CYP1A1 and GSTM1 gene polymorphisms was 8.55 (95%CI: 1.75-41.50). CYP1A1 and GSTM1 polymorphisms are genetic risk factors in patients with bone tumors, and the allelic variation of these genes increases the risk of bone tumor occurrence.

Effect of siRNA targeting HER2/neu on the proliferation and viability of prostate cancer PC-3M cells.

The aim of this study was to investigate the effect of a small interfering RNA (siRNA) targeting human epidermal growth factor receptor 2 (HER2/neu) on the proliferation and viability of prostate cancer PC-3M cells. Chemically synthesized siRNA targeting HER2/neu was transfected into PC-3M cells by using liposomes, and cells transfected with empty liposomes, a negative siRNA sequence, or nothing (untransfected) were used as controls. mRNA and protein levels of HER2/neu were detected using reverse transcription-polymerase chain reaction and western blot, respectively. The inhibitory action of HER2/neu siRNA on the in vitro growth of PC-3M cells was assessed by the cholecystokinin 8 assay and apoptosis was detected using flow cytometry. Cells transfected with HER2/neu siRNA showed decreased mRNA and protein levels of HER2/neu compared to control groups (P < 0.05). The survival rate of PC-3M cells decreased significantly after transfection with HER2/neu siRNA compared to that of untransfected cells (55.39 ± 1.60 and 81.42 ± 0.80%, respectively; P < 0.05). The apoptosis rate in cells transfected with HER2/neu siRNA was quite high (45.60 ± 0.70%) compared to that of blank control, empty liposome, and negative siRNA sequence groups (P < 0.05). In conclusion, siRNA targeting HER2/neu inhibits HER2/neu expression in PC-3M cells, resulting in an inhibition in proliferation and an induction of apoptosis.

Proliferation and differentiation of human osteoblasts from a type 2 diabetic patient in vitro.

We investigated the proliferation and differentiation potential of human osteoblasts from a type 2 diabetic patient in vitro. Human osteoblasts were obtained from a healthy subject and a type 2 diabetic patient and were cultured in vitro using the tissue explant adherent method. Differences in cell morphology were observed under a phase contrast microscope. The osteogenic differentiation capacity was evaluated by alizarin red staining, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, alkaline phosphatase (ALP) staining, and detection of bone Gla-protein (BGP) and Col-1. Expression of Runx-2 and Col-1 was detected using RT-PCR and western blot. Our data indicated that alveolar bone osteoblasts from the type 2 diabetic patient exhibited poorer growth, smaller calcium nodule formation, slower proliferation, and lower ALP, BGP, and Col-1 concentrations in the cell culture supernatant, as compared to control cells (P < 0.05). Combined, our study indicated that alveolar bone osteoblasts from a type 2 diabetic patient exhibited slower proliferation and decreased differentiation, as compared to healthy control, when cultured in vitro.

Quantative trait loci of seed traits for soybean in multiple environments.

Seed length and seed width are an important factor to the soybean yield. So the quantitative trait loci (QTL) location for seed length and seed width could assistant the breeding of soybean. In this study, the QTL underlying seed length and seed width were studied. A recombinant inbred line population of soybeans derived from a cross between the American semi-draft cultivars Charleston and Dongnong 594 were used in 7 environments. The quantitative trait loci underlying seed length, seed width, and seed length/seed width were analyzed by the method of composite interval mapping. Then, the epistatic effects and the QTL-environment (QE) interaction effects were also analyzed. Some valuable QTL sites found had great effect to the seed trait. Results showed that 7 QTLs underlying seed length were identified mainly on linkage groups D1a, C2, B1, A1, G, and A2. For the seed width, 7 QTLs were identified on linkage groups D1a and O. Two QTLs of seed length/seed width were identified on linkage groups D1b and C2. No QE interaction was found for QTLs of seed length and seed width in 7 environments. QTLs of seed length/seed width on linkage groups A1 and I had a QE interaction in 7 environments. Seven pairs of QTLs were identified that affected additive x additive epistatic effect of seed length, seed width, and seed length/seed width, which occurred among 8 linkage groups. These results supply a good foundation for molecular assistant breeding for soybean seed trait.

Fas/FasL in the immune pathogenesis of severe aplastic anemia.

Fas/FasL protein expression of bone marrow hematopoietic cells was investigated in severe aplastic anemia (SAA) patients. Fas expression was evaluated in CD34(+), GlycoA(+), CD33(+), and CD14(+) cells labeled with monoclonal antibodies in newly diagnosed and remission SAA patients along with normal controls. FasL expression was evaluated in CD8(+) cells in the same manner. In CD34(+) cells, Fas expression was significantly higher in the newly diagnosed SAA group (46.59 ± 27.60%) than the remission (6.12 ± 3.35%; P < 0.01) and control (8.89 ± 7.28%; P < 0.01) groups. In CD14(+), CD33(+), and GlycoA(+) cells, Fas levels were significantly lower in the newly diagnosed SAA group (29.29 ± 9.23, 46.88 ± 14.30, and 15.15 ± 9.26%, respectively) than in the remission (47.23 ± 31.56, 67.22 ± 34.68, and 43.56 ± 26.85%, respectively; P < 0.05) and normal control (51.25 ± 38.36, 72.06 ± 39.88, 50.38 ± 39.88%, respectively; P < 0.05) groups. FasL expression of CD8(+) cells was significantly higher in the newly diagnosed SAA group (89.53 ± 45.68%) than the remission (56.39 ± 27.94%; P < 0.01) and control (48.63 ± 27.38%; P <0.01) groups. No significant differences were observed between the remission and control groups. FasL expression in CD8(+) T cells was significantly higher in newly diagnosed patients, and CD34(+), CD33(+), CD14(+), and GlycoA(+) cells all showed Fas antigen expression. The Fas/FasL pathway might play an important role in excessive hematopoietic cell apoptosis in SAA bone marrow. Furthermore, CD34(+) cells are likely the main targets of SAA immune injury.

Effect of bradykinin on renal mesangial cell proliferation and extracellular matrix secretion.

Recent studies have found that bradykinin (BK) plays a role in delaying glomerulosclerosis, although the mechanism of this phenomenon remains unclear. Mesangial cell proliferation (MCP) and extracellular matrix (ECM) secretion are important mechanisms for glomerulosclerosis. This study investigated the impact of BK on the platelet-derived growth factor (PDGF)-induced proliferation of mesangial cells, and evaluated its correlations with the extracellular signal-related kinase (ERK) signaling pathway. The results showed that on its own, 10-1000 mg/L BK promoted MCP and ECM secretion and induced ERK phosphorylation. However, BK administration after PDGF pre-incubation inhibited PDGF-induced MCP, ECM secretion, and ERK phosphorylation. The BK B2 receptor-specific antagonist, HOE-140, and tyrosine phosphatase inhibitor (OV) effectively blocked the function of BK. In summary, these results demonstrated that BK has a bidirectional effect on MCP and ECM secretion: when used alone, it promoted effects on these phenomena, but these effects were inhibited when combined with PDGF. This suggests that the role of BK might be achieved through inhibiting activation of the PDGF-induced ERK1/2 pathway.

Profiles of circulating endothelial cells and serum cytokines during adjuvant chemoradiation in rectal cancer patients.

INTRODUCTION: This research aimed to demonstrate the correlation of circulating endothelial cells (CECs) count and serum cytokine levels with side effects and prognosis in rectal cancer patients receiving adjuvant chemoradiation. METHODS: Eleven patients received proctectomy, chemoradiotherapy and follow-up for 4 years. Fifty-five blood samples were taken before radiation and during the course. The quantities of CECs were estimated by flow cytometry, and serological factors were measured by ELISA. RESULTS: The CEC level in patients without tumor recurrence was significantly lower than in patients with tumor recurrence (p < 0.01). The IL-6 and TGF-ß1 levels exhibited a similar profile (p < 0.01). For morbidity, the mean CEC level in patients with grade 3 diarrhea was significantly greater than patients with grades 1 (p < 0.001) and 2 diarrhea (p < 0.005). CONCLUSIONS: Levels of CECs, serum IL-6, TGF-ß1 and TNF-α during post-operative chemoradiation in rectal cancer patients might be candidate biomarkers for prognosis and morbidity (NCT00325871).

Cutaneous abscess caused by Nocardia brasiliensis: report of a case.

We report a case of cutaneous Nocardia brasiliensis infection. The patient had received radiotherapy and anti-neoplastic chemotherapy for epidermoid carcinoma of the left sphenoid sinus with bone destruction. He developed fever and an ulcer on the dorsal medial surface of the left hand after an intravenous infusion of chemotherapeutic agents in the same site 3 days earlier. Needle aspiration of the abscess disclosed polymorphonuclear leukocytes, and a partially acid-fast, gram-positive filamentous branching organism. Cultures of the aspirate grew N. brasiliensis 1 week later. The patient was treated successfully with a regimen of parenteral ceftazidime and amikacin with definite improvement 1 week later. Therapy was continued for 1 more week, and then the patient was switched to oral trimethoprim-sulfamethoxazole for 3 months with no recurrence. The diagnosis, clinical manifestations, treatment and prognosis of cutaneous abscesses cause by N. brasiliensis are discussed.