Overexpression of Long Non-Coding RNA MEG3 Inhibits Proliferation of Hepatocellular Carcinoma Huh7 Cells via Negative Modulation of miRNA-664.

Evidence is accumulating that long non-coding RNAs (lncRNAs) are involved in human tumorigenesis and dysregulated in many cancers, including hepatocellular carcinoma (HCC). Because lncRNAs can regulate essential pathways that contribute to tumor initiation and progression with their tissue specificity, lncRNAs are valuable biomarkers and therapeutic targets. Maternally expressed gene 3 (MEG3) is a lncRNA overexpressed in HCC cells that inhibits HCC progression, however, the mechanism remains largely unknown. Recently, a novel regulatory mechanism has been proposed in which RNAs can cross-talk with each other via competing for shared microRNAs (miRNAs). The proposed competitive endogenous RNAs could mediate the bioavailability of miRNAs on their targets, thus imposing another level of post-transcriptional regulation. In the current study, we demonstrated that MEG3 is down-regulated in HCC tissues. MEG3 over-expression imposes another level of post-transcriptional regulation, whereas MEG3 overexpression increase the expression of the miR-664 target gene, ADH4, through competitive "sponging" miR-664. In addition, NF-κB may affect transcription of MEG3 by directly binding to the promoter region. Our data revealed that NF-κB may affect the transcript of MEG3. MEG3 overexpression inhibited the proliferation of HCC cells, at least in part by affecting miR-664mediated regulation of ADH4. Together, these results suggest that MEG3 is a suppressor of tumor which acts in part through "sponging" miR-664. J. Cell. Biochem. 118: 3713-3721, 2017. © 2017 Wiley Periodicals, Inc.

Identification of keystone taxa in rhizosphere microbial communities using different methods and their effects on compounds of the host Cinnamomum migao.

Exploring keystone taxa affecting microbial community stability and host function is crucial for understanding ecosystem functions. However, identifying keystone taxa from humongous microbial communities remains challenging. We collected 344 rhizosphere and bulk soil samples from the endangered plant C. migao for 2 years consecutively. Used high-throughput sequencing 16S rDNA and ITS to obtain the composition of bacterial and fungal communities. We explored keystone taxa and the applicability and limitations of five methods (SPEC-OCCU, Zi-Pi, Subnetwork, Betweenness, and Module), as well as the impact of microbial community domain, time series, and rhizosphere boundary on the identification of keystone taxa in the communities. Our results showed that the five methods, identified abundant keystone taxa in rhizosphere and bulk soil microbial communities. However, the keystone taxa shared by the rhizosphere and bulk soil microbial communities over time decreased rapidly decrease in the five methods. Among five methods on the identification of keystone taxa in the rhizosphere community, Module identified 113 taxa, SPEC-OCCU identified 17 taxa, Betweenness identified 3 taxa, Subnetwork identified 3 taxa, and Zi-Pi identified 4 taxa. The keystone taxa are mainly conditionally rare taxa, and their ecological functions include chemoheterotrophy, aerobic chemoheterotrophy, nitrate reduction, and anaerobic photoautotrophy. The results of the random forest model and structural equation model predict that keystone taxa Mortierella and Ellin6513 may have an effects on the accumulation of 1, 4, 7, - Cycloundecatriene, 1, 5, 9, 9-tetramethyl-, Z, Z, Z-, beta-copaene, bicyclogermacrene, 1,8-Cineole in C. migao fruits, but their effects still need further evidence. Our study evidence an unstable microbial community in the bulk soil, and the definition of microbial boundary and ecologically functional affected the identification of keystone taxa in the community. Subnetwork and Module are more in line with the definition of keystone taxa in microbial ecosystems in terms of maintaining community stability and hosting function.

Seed Dormancy Release and Germination Requirements of Cinnamomum migao, an Endangered and Rare Woody Plant in Southwest China.

Seed dormancy is a complex adaptive trait of plants that are influenced by several physiological and environmental factors. The endangered plant Cinnamomum migao is also known to exhibit seed dormancy and low germination, which may influence its regeneration; however, these characteristics remain unexplored. To our knowledge, this study is the first to examine the type of dormancy and improve the germination percentage of C. migao seeds. We evaluated the structure and characteristics of the embryo and endocarp of C. migao seeds as well as the effects of endogenous inhibitors. Furthermore, we assessed the effects of light, stratification, alternating temperature, and gibberellic acid 3 (GA3) on the dormancy release of these seeds. The embryo was well developed the endocarp was water-permeable, and no obvious mechanical hindrance to germination was observed. However, the endocarp and embryo contained phenols and other germination inhibitors. The seed extracts of C. migao delayed the germination of cabbage and ryegrass seeds, which indicates the presence of endogenous inhibitors. These findings suggest that C. migao seeds exhibit physiological dormancy. Light and an alternating temperature (15/20°C) did not influence germination. However, GA3 pretreatment, alternating temperatures, and warm stratification relieved dormancy. GA3 pretreatment combined with the 15°C stratification treatment was most effective in rapidly releasing the C. migao seed dormancy. Our findings may facilitate the storage and conservation of this endangered plant, which is currently underrepresented in ex situ collections.

Simulated atmospheric nitrogen deposition inhibited the leaf litter decomposition of Cinnamomum migao H. W. Li in Southwest China.

Atmospheric nitrogen (N) deposition could affect various ecological processes in forest ecosystems, including plant litter decomposition and nutrient cycling. However, the mechanism of underlying litter decomposition and nutrient cycling of Cinnamomum migao under N deposition remains unclear. Therefore, we conducted a simulated N deposition experiment including four onsite treatments to assess the effects of N input on C. migao leaf litter decomposition, nutrient release, and soil enzyme activity. The results showed that simulated N deposition significantly increased the amount of total residual mass and lignin and cellulose, decreased the decomposition rate, and suppressed net nutrient release. N input increased C, N, and P ratios as decomposition progressed, and the proportion of mass remaining was positively correlated with the proportions of lignin and cellulose remaining at the later stage of decomposition. The differences in soil enzyme activity were primarily due to enzyme type and sampling time. We conclude that simulated N deposition significantly suppressed the leaf litter decomposition of C. migao by mainly altering the chemical properties and suppressing the decomposition of the organic matter in leaf litter. Lignin might have played an important role in the loss of leaf litter biomass at the later stage of decomposition.

Clonal integration affects growth, photosynthetic efficiency and biomass allocation, but not the competitive ability, of the alien invasive Alternanthera philoxeroides under severe stress.

BACKGROUND AND AIMS: Many notorious alien invasive plants are clonal, but little is known about some roles and aspects of clonal integration. Here, the hypothesis is tested that clonal integration affects growth, photosynthetic efficiency, biomass allocation and competitive ability of the exotic invasive weed Alternanthera philoxeroides (Amaranthaceae). METHODS: The apical parts of Alternanthera were grown either with or without the lawn grass Schedonorus phoenix (tall fescue) and their stolon connections to the basal parts grown without competitors were either severed or left intact. KEY RESULTS: Competition greatly reduced the maximum quantum yield of photosystem II (F(v)/F(m)) and growth (biomass, number of ramets and leaves, total stolon length and total leaf area) of the apical Alternanthera, but not the biomass of S. phoenix. Stolon connections significantly increased F(v)/F(m) and growth of Alternanthera. However, such effects on growth were smaller with than without competition and stolon connections did not alter the relative neighbour effect of Alternanthera. Stolon connections increased Alternanthera's biomass allocation to roots without competition, but decreased it with competition. CONCLUSIONS: Clonal integration contributed little to Alternanthera's competitive ability, but was very important for Alternanthera to explore open space. The results suggest that the invasiveness of Alternanthera may be closely related to clonal integration.

UNBS5162 inhibits colon cancer growth via suppression of PI3K/Akt signaling pathway.

Colon cancer is a common cause of cancer-related death worldwide. However, the underlying mechanism of tumor progression of colon cancer remains far from being elucidated. In the present study, we report the role of UNBS5162 in colon cancer. UNBS5162 is a naphthalimide that can intercalate into DNA and suppress the expression level of CXCL chemokines. Here, we investigated its effect on cell proliferation, mobility and apoptosis in HCT116 cells, and explored the underlying mechanism. A CCK8 assay revealed that UNBS5162 can block the proliferation of colon cancer cells. Base on a Transwell assay, we showed that cell migration and invasion ability of HCT116 cells are inhibited by UNBS5162. In addition, Annexin V-FITC/PI assay and Western blot analysis were performed to detect whether UNBS5162 could induce cell apoptosis. The results indicated that UNBS5162 increases the number of apoptotic cells remarkably. Furthermore, Western blot analysis demonstrated that UNBS5162 down-regulates the expression level of Bcl2, and up-regulates that of Bax as well as the level of activated Caspase-3. Moreover, we examined the impact of UNBS5162 on PI3K/Akt signaling pathway. UNBS5162 substantially inhibited the phosphorylation of Akt and its downstream effector mTOR, and reduced the expression of p-70. Taken together, these results suggest that UNBS5162 should be considered as a potent therapeutic anticancer agent that targets the PI3K/AKT signaling pathway.

Bioinformatic analysis and prediction of the function and regulatory network of long non-coding RNAs in hepatocellular carcinoma.

Computational analysis and bioinformatics have significantly advanced the ability of researchers to process and analyze biological data. Molecular data from human and model organisms may facilitate drug target validation and identification of biomarkers with increased predictive accuracy. The aim of the present study was to investigate the function of long non-coding RNAs (lncRNAs) in hepatocellular carcinoma (HCC) using online databases, and to predict their regulatory mechanism. HCC-associated lncRNAs, their downstream transcription factors and microRNAs (miRNAs/miRs), as well as the HCC-associated target genes, were identified using online databases. HCC-associated lncRNAs, including HOX antisense intergenic RNA (HOTAIR) and metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) were selected based on established databases of lncRNAs. The interaction between the HCC-associated lncRNAs and miRNAs (hsa-miR-1, hsa-miR-20a-5p) was predicted using starBase2.0. Signal transducer and activator of transcription 1, hepatocyte nuclear factor 4α (HNF4A), octamer-binding transcription factor 4, Nanog homeobox (NANOG), caudal type homeobox 2 (CDX2), DEAD-box helicase 5, brahma-related gene 1, MYC-associated factor X and MYC proto-oncogene, bHLH transcription factor have been identified as the transcription factors for HOTAIR and MALAT1 using ChIPBase. Additionally, CDX2, HNF4A, NANOG, ETS transcription factor, Jun proto-oncogene and forkhead box protein A1 were identified as the transcription factors for hsa-miR-1 and hsa-miR-20a-5p. CDX2, HNF4A and NANOG were the transcriptional factors in common between the lncRNAs and miRNAs. Cyclin D1, E2F transcription factor 1, epithelial growth factor receptor, MYC, MET proto-oncogene, receptor tyrosine kinase and vascular endothelial growth factor A were identified as target genes for the HCC progression, two of which were also the target genes of hsa-miR-1 and hsa-miR-20a-5p using the miRwalk and OncoDN. HCC databases. Additionally, these target genes may be involved in biological functions, including the regulation of cell growth, cell cycle progression and mitosis, and in disease progression, as demonstrated using DAVID clustering analysis. The present study aimed to predict a regulatory network of lncRNAs in HCC progression using bioinformatics analysis.

[Clinical observation on effect of electric acupuncture at Sishencong in treating insomnia].

OBJECTIVE: To evaluate the clinical therapeutic effect of electric acupuncture (EA) at Sishencong (EX-HN 1) on insomnia. METHODS: Two hundred and seventy-six patients were randomly assigned to 2 groups, 138 in each group, the EA group treated with EA at Sishencong, and the control group with oral administration of Tianmeng Capsule. The treatment course for both groups was 3 weeks. The quality and related parameters of sleep before and after treatment were evaluated with a multi-channel sleep detector. RESULTS: After treatment, the quality of sleep was improved in both groups (P < 0.05), as compared with before treatment, the difference in related parameters was significant respectively (P<0.05 or P <0.01), however, the improvement in the EA group was superior to that in the control group (P < 0.01). CONCLUSION: EA at Sishencong has obvious effect on insomnia.

Biomechanics of anterior plating failure in treating distractive flexion injury in the caudal subaxial cervical spine.

BACKGROUND: Operative level is a potential biomechanical risk factor for construct failure during anterior fixation for distractive flexion injury. No biomechanical study of this concept has been reported, although it is important in clinical management. METHODS: To explore the mechanism of this concept, a previously validated three-dimensional C2-T1 finite element model was modified to simulate surgical procedure via the anterior approach for treating single-level distractive flexion injury, from C2-C3 to C7-T1. Four loading conditions were used including no-compression, follower load, axial load, and combined load. Construct stability at the operative level was assessed. FINDINGS: Under these loading conditions with the head's weight simulated, segmental stability decreases when the operative level shifts cephalocaudally, especially at C6-C7 and C7-T1, the stress of screw-bone interface increases cephalocaudally, and in the same operative level, the caudal screws always carries more load than the cephalad ones. All these predicted results are consistent with failure patterns observed in clinical reports. In the contrast, under other loading conditions without the weight of head, no obvious segmental divergence was predicted. INTERPRETATION: This study supports that the biomechanical mechanism of this phenomenon includes eccentric load from head weight during sagittal movements and difference of moment arms. Our study suggests that anterior fixation is not recommended for treating distractive flexion injury at the caudal segments of the subaxial cervical spine, especially at C6-C7 and C7-T1, because of the intrinsic instability in these segments. Combined posterior rigid fixation with anterior fixation should be considered for these segments.

Stochastic modelling of infectious diseases for heterogeneous populations.

BACKGROUND: Infectious diseases such as SARS and H1N1 can significantly impact people's lives and cause severe social and economic damages. Recent outbreaks have stressed the urgency of effective research on the dynamics of infectious disease spread. However, it is difficult to predict when and where outbreaks may emerge and how infectious diseases spread because many factors affect their transmission, and some of them may be unknown. METHODS: One feasible means to promptly detect an outbreak and track the progress of disease spread is to implement surveillance systems in regional or national health and medical centres. The accumulated surveillance data, including temporal, spatial, clinical, and demographic information can provide valuable information that can be exploited to better understand and model the dynamics of infectious disease spread. The aim of this work is to develop and empirically evaluate a stochastic model that allows the investigation of transmission patterns of infectious diseases in heterogeneous populations. RESULTS: We test the proposed model on simulation data and apply it to the surveillance data from the 2009 H1N1 pandemic in Hong Kong. In the simulation experiment, our model achieves high accuracy in parameter estimation (less than 10.0 % mean absolute percentage error). In terms of the forward prediction of case incidence, the mean absolute percentage errors are 17.3 % for the simulation experiment and 20.0 % for the experiment on the real surveillance data. CONCLUSION: We propose a stochastic model to study the dynamics of infectious disease spread in heterogeneous populations from temporal-spatial surveillance data. The proposed model is evaluated using both simulated data and the real data from the 2009 H1N1 epidemic in Hong Kong and achieves acceptable prediction accuracy. We believe that our model can provide valuable insights for public health authorities to predict the effect of disease spread and analyse its underlying factors and to guide new control efforts.

Resection or degeneration of uncovertebral joints altered the segmental kinematics and load-sharing pattern of subaxial cervical spine: A biomechanical investigation using a C2-T1 finite element model.

The uncovertebral joint (UJ) is an important load-bearing structure in the subaxial cervical spine (SCS) and the medial wall of the intervertebral foramen (IVF). To investigate the UJ׳s role in load distribution and transmission under physiological loading, we developed and validated a detailed finite element model (C2-T1). Based on the initial model, two additional models were modified to simulate surgical resection and degeneration of UJs, to evaluate their influence on SCS kinematics and load distribution. The three models were subjected to 2Nm pure moment (flexion, extension, lateral bending, and axial rotation). Foraminal narrowing and potential nerve compression were evaluated. In the initial model, contact forces provided by the UJ were apparent in lateral bending and axial rotation. In axial rotation, the UJs and contralateral facet joints participated in joint activity, implying a possible restraint/counterbalance mechanism of these two joints. Peak vertebral stress was observed in the pedicle of vertebrae and was higher in the uncovertebral region than in the facet region. Resection of uncinate processes led to an apparent range of motion increase in lateral bending and axial rotation, while sagittal kinematics is influenced slightly. The load on other structures was slightly increased, but in axial rotation, resection of UJs changed the load distribution pattern. Degeneration of UJs significantly increased SCS stiffness and shielded other load-bearing structures. Peak IVF narrowing, but no nerve compression, was observed in axial rotation of the resection model. Thus, resection did not induce apparent secondary foraminal stenosis when other structures were still functional.

[Growth response of Broussonetia papyrifera seedlings to VA mycorrhizal fungi inoculation].

In an experiment with single inoculation (SI) and co-inoculation (CI) of three VA mycorrhizal fungi, i. e., Glomus mosseea (GM), Glomus versiforme (GV) and Glomus diaphanum (GD), the growth response of Broussonetia papyrifera seedlings in limestone area was studied. The results showed that after 3 months of growth, the aboveground-, underground-, and total biomass were increased significantly by the inoculation, being 2.49-8.19 times as much as the control. Treatment CI had the highest biomass, but the leaf number had little difference with the control. In SI, GD inoculation had the greatest effect; and CI was more effective than SI. The ground diameter, height, and total leaf area in CI were 1.5, 2.2, and 6.0 times as much as those in CK, respectively, and the root/shoot ratio in CI was the highest (0.446). There existed an interactive selection between host plants and VA mycorrhizal fungi.

[Evaluation of therapeutic effects of synthetic auricular point therapy for treatment of common migraine at the attack stage].

OBJECTIVE: To observe and evaluate the short-term and long-term curative effects of the synthetic auricular point therapy on the common migraine in attack stage. METHODS: With the study method of randomized controlled trial, 108 cases of common migraine at the attack stage were randomly assigned to a treatment group of 72 cases treated with synthetic auricular point therapy (blood-letting on the ear back, point injection of own blood, pricking ear point), and a control group of 36 cases treated with Western medicine. Their short-term and long-term curative effects were observed and evaluated. RESULTS: The total short-term effective rate and the total long-term effective rate were respectively 98.6% and 79. 6% in the treatment group with less adverse effects, and with a very significant difference as compared with the control group (P < 0.01). CONCLUSION: The synthetic auricular point therapy has obvious short-term and long-term curative effects on common migraine at the attack stage with safety.

[Effect of acupuncture at Sishencong (EX-HN 1) on sleeping in the patient of insomnia].

OBJECTIVE: To assess therapeutic effect of acupuncture at Sishencong (EX-HN 1) on insomnia. METHODS: Sixty-five cases of insomnia were treated with acupuncture at Sishencong (EX-HN 1) and their quality and compositions of sleeping before and after treatment were evaluated with a multichannel sleep detector. RESULTS: After treatment, the quality of sleeping increased significantly (P < 0.05), and the sleeping rate increased from (61.95 +/- 22.90)% before treatment to (79.27 +/- 11.05)% after treatment (P < 0.05). CONCLUSION: Acupuncture at Sishencong (EX-HN 1) has obvious therapeutic effect on insomnia.

[Expression of CXCR4 and its effect on the biological behavior of multiple myeloma cells].

OBJECTIVE: To investigate the effect of stromal cell derived factor-1alpha(SDF-1alpha) expression and its receptor CXCR4 on the biological behavior of multiple myeloma (MM) cells and on the expression of soluble intercellular adhesion molecule 1 (ICAM-1). METHODS: FACS analysis was used to study the expression of ICAM-1 (CD(54)) and CXCR4 on the surface of MM cells. Chemotaxis assay through transwell bore polycaronate and ELISA assay were employed to monitor the soluble ICAM-1 level. RESULTS: (1) Fresh MM cells expressed variable levels of functional CXCR4 [(50.4 +/- 27.3)%], which was correlated with the in vitro ability of transwell migration of MM cells [(23.6 +/- 17.2)%, P < 0.01]. (2) SDF-1alpha could up-regulate the expression of ICAM-1 on MM cells. Furthermore, the serum level of sICAM-1 was correlated with the expression of CXCR4 on MM cells. CONCLUSION: SDF-1alpha/CXCR4 plays an important role on the biological behavior of MM cells via mediating the effect of adhesion molecules.