Network pharmacology-based approach to investigate the mechanisms of Zingiber officinale Roscoe in the treatment of neurodegenerative diseases.

Neurodegenerative diseases (NDDs) are chronic neurological disorders associated with cognitive or motor dysfunction. As a common spice, Zingiber officinale Roscoe has been used as a medicine to treat a variety of NDDs. However, at the molecular level, the mechanisms of Z. officinale in treating of NDDs have not been deeply investigated. In this study, network pharmacology method, molecular docking, Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis were used to predict the mechanisms of Z. officinale in the treatment of NDDs. After a series of biological information analyses, five core targets were obtained, including heme oxygenase 1 (HMOX1), acetylcholinesterase (AChE), nitric oxide synthase (NOS), catechol-O-methyl-transferase (COMT), and metabotropic glutamate receptor 5 (mGluR5). Compounds 75, 68, 46, 67, 69, 49, 66, 50, 34, and 64 were identified as the main components of Z. officinale in the treatment of NDDs. The crucial pathways mainly include neuroactive ligand-receptor signaling pathways, cyclic adenosine monophosphate signaling pathways, dopamine synaptic signaling pathways, and so on. Besides, in vitro experiments by AChE inhibitory activities assay and neuroprotective activities against H2 O2 -induced injury in human neuroblastoma SH-SY5Y cells validated the reliability of the results of network analysis. PRACTICAL APPLICATIONS: Zingiber officinale Roscoe is widely used as a traditional spice and herbal medicine. It contains a number of active ingredients, which have shown activities on anti-neurodegenerative diseases (NDDs). In this paper, the potential mechanism of Z. officinale in the treatment of NDDs is explored through network pharmacology, and it was verified by in vitro experiments. The mechanism was not only clarified at the system level but also proved to be effective at the biological level. The results can be used as a reference for Z. officinale in the treating of NDDs.

ROS/p53/miR­335­5p/Sp1 axis modulates the migration and epithelial to mesenchymal transition of JEG­3 cells.

Differential expression of microRNA (miR)­335­5p, a key tumor suppressor, has been detected in pre­eclampsia (PE) placentas. However, the role of miR­335­5p in the pathogenesis of PE and the factor modulating its aberrant expression remain unknown. The present study used JEG­3 cells in vitro to investigate these mechanisms. The role of miR­335­5p in proliferation, apoptosis and migration of JEG­3 cells was investigated using MTT, Annexin V­FITC/PI, Transwell migration and wound healing assays, respectively. miR­335­5p expression levels were analyzed using reverse transcription­quantitative PCR. The expression levels of E­cadherin, N­cadherin, Snail, specificity protein 1 (Sp1) and p53 were assessed using western blot analysis. Cell viability analysis was performed using the Cell Counting Kit­8 assay. The intracellular reactive oxygen species (ROS) levels were detected using a 2,7­dichlorodihydrofluorescein diacetate assay. The present results suggested that miR­335­5p did not affect the proliferation or apoptotic rate of JEG­3 cells. Overexpression of miR­335­5p significantly inhibited the migration of JEG­3 cells, decreased the expression levels of Sp1, N­cadherin and Snail, and increased E­cadherin expression. Sp1 silencing produced similar results in JEG­3 cells. H2O2 significantly increased the intracellular ROS levels and miR­335­5p expression, whereas N­acetyl­cysteine pretreatment prior to H2O2 treatment reversed the increases in miR­335­5p expression. Knockdown of p53 significantly decreased the expression levels of miR­335­5p in JEG­3 cells and in H2O2­treated cells. The present results suggested that miR­335­5p expression levels in trophoblast cells could be increased by ROS in a p53­dependent manner, leading to the downregulation of Sp1 and subsequent inhibition of epithelial to mesenchymal transition and cell migration. The present results may provide novel evidence on the etiology of PE.

Ubiquitination-Related MdBT Scaffold Proteins Target a bHLH Transcription Factor for Iron Homeostasis.

Iron (Fe) homeostasis is crucial for plant growth and development. A network of basic helix-loop-helix (bHLH) transcription factors positively regulates Fe uptake during iron deficiency. However, their up-regulation or overexpression leads to Fe overload and reactive oxygen species generation, thereby damaging the plants. Here, we found that two BTB/TAZ proteins, MdBT1 and MdBT2, interact with the MbHLH104 protein in apple. In addition, the function of MdBT2 was characterized as a regulator of MdbHLH104 degradation via ubiquitination and the 26S proteasome pathway, thereby controlling the activity of plasma membrane H+-ATPases and the acquisition of iron. Furthermore, MdBT2 interacted with MdCUL3 proteins, which were required for the MdBT2-mediated ubiquitination modification of MdbHLH104 and its degradation. In sum, our findings demonstrate that MdBT proteins interact with MdCUL3 to bridge the formation of the MdBTsMdCUL3 complex, which negatively modulates the degradation of the MdbHLH104 protein in response to changes in Fe status to maintain iron homeostasis in plants.

The enhancement of tolerance to salt and cold stresses by modifying the redox state and salicylic acid content via the cytosolic malate dehydrogenase gene in transgenic apple plants.

In this study, we characterized the role of an apple cytosolic malate dehydrogenase gene (MdcyMDH) in the tolerance to salt and cold stresses and investigated its regulation mechanism in stress tolerance. The MdcyMDH transcript was induced by mild cold and salt treatments, and MdcyMDH-overexpressing apple plants possessed improved cold and salt tolerance compared to wild-type (WT) plants. A digital gene expression tag profiling analysis revealed that MdcyMDH overexpression largely altered some biological processes, including hormone signal transduction, photosynthesis, citrate cycle and oxidation-reduction. Further experiments verified that MdcyMDH overexpression modified the mitochondrial and chloroplast metabolisms and elevated the level of reducing power, primarily caused by increased ascorbate and glutathione, as well as the increased ratios of ascorbate/dehydroascorbate and glutathione/glutathione disulphide, under normal and especially stress conditions. Concurrently, the transgenic plants produced a high H2 O2 content, but a low O2·- production rate was observed compared to the WT plants. On the other hand, the transgenic plants accumulated more free and total salicylic acid (SA) than the WT plants under normal and stress conditions. Taken together, MdcyMDH conferred the transgenic apple plants a higher stress tolerance by producing more reductive redox states and increasing the SA level; MdcyMDH could serve as a target gene to genetically engineer salt- and cold-tolerant trees.

Overexpression of MdbHLH104 gene enhances the tolerance to iron deficiency in apple.

Fe deficiency is a widespread nutritional disorder in plants. The basic helix-loop-helix (bHLH) transcription factors (TFs), especially Ib subgroup bHLH TFs which are involved in iron uptake, have been identified. In this study, an IVc subgroup bHLH TF MdbHLH104 was identified and characterized as a key component in the response to Fe deficiency in apple. The overexpression of the MdbHLH104 gene noticeably increased the H(+) -ATPase activity under iron limitation conditions and the tolerance to Fe deficiency in transgenic apple plants and calli. Further investigation showed that MdbHLH104 proteins bonded directly to the promoter of the MdAHA8 gene, thereby positively regulating its expression, the plasma membrane (PM) H(+) -ATPase activity and Fe uptake. Similarly, MdbHLH104 directly modulated the expression of three Fe-responsive bHLH genes, MdbHLH38, MdbHLH39 and MdPYE. In addition, MdbHLH104 interacted with 5 other IVc subgroup bHLH proteins to coregulate the expression of the MdAHA8 gene, the activity of PM H(+) -ATPase and the content of Fe in apple calli. Therefore, MdbHLH104 acts together with other apple bHLH TFs to regulate Fe uptake by modulating the expression of the MdAHA8 gene and the activity of PM H(+) -ATPase in apple.

The 5-hydroxytryptamine transporter is functional in human coronary artery smooth muscle cells proliferation and is regulated by Interleukin-1 beta.

Abnormal human coronary artery smooth muscle cells (hCASMCs) proliferation and migration are key factors in coronary artery restenosis after percutaneous coronary intervention. Platelets release 5-hydroxytryptamine (5-HT), which is a strong mitogen for pulmonary artery smooth muscle cells proliferation and migration. Here, we investigated the effects of 5-HT and role of 5-HT transporter (5-HTT) on hCASMCs proliferation and migration. The 5-HT (10(-6)-10(-5) mol/l) significantly increased hCASMCs proliferation and migration, and these effects were inhibited by fluoxetine (10(-5) mol/l) and citalopram (10(-6) mol/l), two 5-HTT blocker. Overexpression in hCASMCs enhanced 5-HT induced cells proliferation and migration. The 5-HTT and interleukin-1 beta (IL-1ß) expression were increased in rat balloon injury carotid arteries. Treatment with IL-1ß (10 ng/ml, 3d) upregulates 5-HTT expression in hCASMCs and increased 5-HT induced currents in Human Embryonic Kidney 293-5-HTT cells.

Overexpression of chemerin was associated with tumor angiogenesis and poor clinical outcome in squamous cell carcinoma of the oral tongue.

OBJECTIVE: The present study aimed to explore expression and clinical significance of chemerin, a newly discovered adipokine, in squamous cell carcinoma of the oral tongue (SCCOT). METHODS: mRNA expression of chemerin in 19 pairs of fresh SCCOT samples matched with peritumoral mucosa tissues was quantified by real-time quantitative transcription polymerase chain reaction (qRT-PCR). Chemerin protein expression and microvessel density (MVD) were measured by immunohistochemistry on 147 cases of primary SCCOT specimen and their corresponding peritumoral noncancerous tissues. The relationship of chemerin expression with angiogenesis, clinicopathologic parameters, and cancer-related survival of patients was evaluated. RESULTS: Both qRT-PCR and immunohistochemistry results revealed that chemerin was overexpressed in SCCOT compared with peritumoral noncancerous tissues (P < 0.01). Overexpression of chemerin in SCCOT was significantly associated with poor differentiation, lymph node metastasis, and high clinical stage (P = 0.000, 0.012, and 0.015, respectively). In addition, overexpression of chemerin was positively related to MVD in SCCOT (r = 0.671, P = 0.002). SCCOT patients with overexpressed chemerin had a shorter cancer-related survival (P = 0.027). Moreover, multivariate survival analysis indicated that chemerin was an independent prognostic factor for SCCOT patients (P = 0.016). CONCLUSION: These results demonstrated that overexpression of chemerin in SCCOT was correlated with tumor angiogenesis and poor clinical outcomes of SCCOT patients. CLINICAL RELEVANCE: Our research implied that chemerin was a novel prognostic factor for SCCOT patients, and chemerin could be a new therapeutic target for regulating tumor angiogenesis and blocking tumor progression.

Expansion of decidual CD45RO⁺ T cells with high expression of CEACAM1 in the early stage of pregnancy.

The aim of the present study was to investigate the mechanism involved in the expansion of CD45RO+ T cells in the decidual microenvironment, and in the expression of the inhibitory carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) on the surface of decidual CD45RO+ T cells. Twenty-one healthy nonpregnant females and seventeen healthy pregnant females in the first trimester were included in the study. Peripheral blood samples from nonpregnant and pregnant females, and decidual tissues from pregnant females following elective abortion, were obtained and analyzed by flow cytometry. The percentages of CD45RO+ T cells and CEACAM1-expressing CD45RO+ T cells were significantly higher in first trimester human decidua than in the peripheral blood. Conditioned medium from the coculture of monocytes and the human trophoblast HTR8/SVneo cell line (MHM) was added to the model for the generation of CD45RO+ T cells in vitro. MHM caused an increase in the percentage of CD45RO+ T cells in a monocyte chemoattractant protein-1 (MCP-1)­dependent manner and an increase in the percentage of CEACAM1-expressing CD4+CD45RO+ T cells in the model. In conclusion, our results implied that trophoblast cells and monocytes may be involved in the increase of decidual CD45RO+ T cells and the high expression of CEACAM1 on their surfaces.

Long-term effects of bone marrow-derived cells transplantation in patients with acute myocardial infarction: a meta-analysis.

BACKGROUND: The long-term effects of bone marrow-derived cells (BMC) transplantation in patients with acute myocardial infarction (AMI) have not been established. The present meta-analysis of randomized controlled trials with follow-up ≥ 2 years was performed to investigate the long-term effects of BMC therapy in patients after AMI. METHODS: Specific terms were used to conduct a systematic literature search of MEDLINE, EMBASE, the Cochrane Library and the Cochrane Central Register of Controlled Trials, and the China Biological Medicine Disk database from their inception to March 2012. A standardized protocol was used to extract information, and random effect model was used to analyze all data except major adverse events. RESULTS: Five trials comprising 510 patients were included. Compared with controls, BMC therapy significantly improved left ventricular ejection fraction (LVEF) (4.18%, 95%CI: 2.02% to 6.35%, P = 0.0002), while mildly but not significantly reduced left ventricular end-systolic volume (-4.47 ml, 95%CI: -10.92 to 1.99, P = 0.17) and left ventricular end-diastolic volume (-2.29 ml, 95%CI: -9.96 to 5.39, P = 0.56). Subgroup analysis revealed that significant improvement of LVEF induced by BMC therapy could be observed in patients with baseline LVEF ≤ 42%, but disappeared in those with baseline LVEF > 42%. There were trends in favor of BMC therapy for most major clinical adverse events, though most differences were not significant. CONCLUSIONS: Intracoronary BMC infusion in patients with AMI seems to be safe and may further improve LVEF on top of standard therapy; especially the beneficial effects could last for long term. The findings need to be validated in the future.

Aberrant expression of chemokine receptor CCR4 in human gastric cancer contributes to tumor-induced immunosuppression.

The chemokine receptor CCR4 is preferentially expressed on certain immune cells and some hematological tumor cells, which play pivotal roles in suppression of host immune response. However, the reasons for the upmodulation of CCR4 and its immune functions in solid tumors remain unclear. Herein, we aimed to determine the expression profiles of CCR4 in gastric cancer cells and its role in regulating antitumor immunity. CCR4 expression was assessed in 63 cases of gastric carcinomas by immunohistochemistry. We found cancer cells in lymphocyte-rich carcinomas more frequently showed moderate to strong positive staining for CCR4 than those in conventional carcinomas (P = 0.041), and also found a positive relationship between expression of CCR4 and tumor necrosis factor-α (P = 0.012). Stimulation of gastric cell lines with various cytokines showed that tumor necrosis factor-α uniquely upmodulated CCR4 expression through activation of nuclear factor-κB. Additional coculture experiments showed the forced expression of CCR4 in SGC-7901 cells caused a significant reduction of γ-interferon and elevation of interleukin-10 secretion in the supernatants from cocultured SGC-7901 cells and PBMCs. In addition, granzyme A production in cancer cell-cocultured CD56(+) natural killer cells was significantly downregulated. Inhibition of the overexpressed CCR4 in cancer cells by an inhibitor of CCR4, compound 39, proved to partly restore the antitumor immunity in respect of the inverse changes in those factors. Our studies suggest that the aberrant expression of CCR4 in human gastric cancer could contribute to tumor-induced immunosuppression. Conceivably, downmodulation of CCR4 expression could be a promising immunotherapy for human gastric cancer.

The adoption of conservation tillage in China.

Conservation tillage (CT) has been recognized as an advanced agricultural technology that may reduce drought and improve the physical condition of soils worldwide. An increase in water infiltration and a reduction in water and wind erosion can be achieved through the use of no-tillage, minimum tillage, and residue cover. In China, CT research started with support from the Ministry of Agriculture (MOA), China and the Australian Centre for International Agricultural Research in 1992. By the end of 1990s, CT research had expanded across China and achieved several important results. In 1999, MOA established the Conservation Tillage Research Centre (CTRC) to lead the national CT research programs in China, and since 2002 some CT demonstration projects have been established in northern China. By the end of 2008, CT has been demonstrated in 226 national and 365 provincial demonstration counties, covering more than 3 Mha. The CTRC of the MOA has established 10 sites within those counties to monitor project results. Some sites have shown consistently that the use of CT resulted in higher yields and net incomes, reduced soil erosion, and improved soil conditions. CT has been widely accepted in China and will be further adopted over wider areas as the development and highbred of indigenous no-tillage seeders.

Dishevelled promotes axon differentiation by regulating atypical protein kinase C.

The atypical protein kinase C (aPKC) in complex with PAR3 and PAR6 is required for axon-dendrite differentiation, but the upstream factors responsible for regulating its activity are largely unknown. Here, we report that in cultured hippocampal neurons aPKC is directly regulated by Dishevelled (Dvl), an immediate downstream effector of Wnt. We found that downregulation of Dvl abrogated axon differentiation, whereas Dvl overexpression resulted in multiple axon formation. Interestingly, Dvl was associated with aPKC and this interaction resulted in aPKC stabilization and activation. Furthermore, the multiple axon formation resulting from Dvl overexpression was attenuated by expressing a dominant-negative aPKC in these neurons and overexpression of aPKC prevented the loss of axon caused by Dvl downregulation. Finally, Wnt5a, a noncanonical Wnt, activated aPKC and promoted axon differentiation. The Wnt5a effect on axon differentiation was attenuated by downregulating Dvl or inhibiting aPKC. Thus, Dvl-aPKC interaction can promote axon differentiation mediated by the PAR3-PAR6-aPKC complex.