Down-regulation of microRNA-203a suppresses IL-1ß-induced inflammation and cartilage degradation in human chondrocytes through Smad3 signaling.

Osteoarthritis (OA) is a chronic and prevalent degenerative musculoskeletal disorder, which is characterized by articular cartilage degradation and joint inflammation. MicroRNA-203a (miR-203a) has been shown to be involved in multiple pathological processes during OA, but little is known about its function in chondrocyte extracellular matrix (ECM) degradation. In the present study, we aimed to elucidate the effects of miR-203a on articular cartilage degradation and joint inflammation. We observed that the miR-203a level was significantly up-regulated in OA tissues and in an in vitro model of OA, respectively. Inhibition of miR-203a significantly alleviated the interleukin (IL)-1ß-induced inflammatory response and ECM degradation in chondrocytes. Moreover, mothers against decapentaplegic homolog 3 (Smad3), a key factor in maintaining chondrocyte homeostasis, was identified as a putative target of miR-203a in chondrocytes. More importantly, inhibition of Smad3 impaired the inhibitory effects of the miR-203a on IL-1ß-induced inflammatory response and ECM degradation. Collectively, these results demonstrated that miR-203a may contribute to articular cartilage degradation of OA by targeting Smad3, suggesting a novel therapeutic target for the treatment of OA.

MiR-125b Inhibits LPS-Induced Inflammatory Injury via Targeting MIP-1α in Chondrogenic Cell ATDC5.

BACKGROUND/AIMS: Chondrocyte apoptosis is largely responsible for cartilage degeneration in osteoarthritis (OA). MicroRNAs (miRNAs) play an important role in chondrogenesis and cartilage remodeling. This study explored the effect of miR-125b on inflammatory injury in chondrogenic cells. METHODS: LPS was used to simulate inflammatory injury in murine chondrogenic ATDC5 cell lines. Targeting effect of miR-125b on MIP-1α 3'UTR was assessed by dual luciferase activity assay. Regulatory effect of miR-125b on MIP-1α expression and the potential regulatory mechanism on inflammatory injury were assessed by Western blot. RESULTS: miR-125b expression was decreased in LPS-induced ATDC5 cells and overexpression of miR-125b inhibited LPS-induced cell viability decline, the rise of apoptosis and inflammatory factors' productions. MIP-1α expression was negatively related to miR-125b, and miR-125b directly targeted with 3'UTR of MIP-1α. Knockdown of miR-125b promoted LPS-induced inflammatory response via upregulation of MIP-1α. miR-125b expression in LPS-induced ATDC5 cells was negatively related with activations of NF-κB and JNK signaling pathways. Overexpression of miR-125b inhibited LPS-induced inflammation injury via suppressing MIP-1α expression and inhibiting activations of NF-κB and JNK signaling pathways. CONCLUSION: miR-125b could play an important role in inflammatory injury of chondrogenic cells and miR-125b affected inflammatory injury of ATDC5 cells via regulating expression of MIP-1α and regulating NF-κB and JNK signaling pathways.

Vulnerability of photosynthesis and photosystem I in Jerusalem artichoke (Helianthus tuberosus L.) exposed to waterlogging.

Jerusalem artichoke (Helianthus tuberosus L.) is an important energy crop for utilizing coastal marginal land. This study was to investigate waterlogging tolerance of Jerusalem artichoke through photosynthetic diagnose with emphasis on photosystem II (PSII) and photosystem I (PSI) performance. Potted plants were subjected to severe (liquid level 5 cm above vermiculite surface) and moderate (liquid level 5 cm below vermiculite surface) waterlogging for 9 days. Large decreased photosynthetic rate suggested photosynthesis vulnerability upon waterlogging. After 7 days of severe waterlogging, PSII and PSI photoinhibition arose, indicated by significant decrease in the maximal photochemical efficiency of PSII (Fv/Fm) and PSI (△MR/MR0), and PSI seemed more vulnerable because of greater decrease in △MR/MR0 than Fv/Fm. In line with decreased △MR/MR0 and unchanged Fv/Fm after 9 days of moderate waterlogging, the amount of PSI reaction center protein rather than PSII reaction center protein was lowered, confirming greater PSI vulnerability. According to positive correlation between △MR/MR0 and efficiency that an electron moves beyond primary quinone and negative correlation between △MR/MR0 and PSII excitation pressure, PSI inactivation elevated PSII excitation pressure by depressing electron transport at PSII acceptor side. Thus, PSI vulnerability induced PSII photoinhibition and endangered the stability of whole photosynthetic apparatus under waterlogging. In agreement with photosystems photoinhibition, elevated H2O2 concentration and lipid peroxidation in the leaves corroborated waterlogging-induced oxidative stress. In conclusion, Jerusalem artichoke is a waterlogging sensitive species in terms of photosynthesis and PSI vulnerability. Consistently, tuber yield was tremendously reduced by waterlogging, confirming waterlogging sensitivity of Jerusalem artichoke.

MiR-1246 Promotes LPS-Induced Inflammatory Injury in Chondrogenic Cells ATDC5 by Targeting HNF4γ.

BACKGROUND/AIMS: Osteoarthritis (OA) is a common inflammatory joint disease. miRNAs are associated with OA and functionally implicated in the pathogenesis of the disease. In the present study, we investigated the role of miR-1246 in the lipopolysaccharide (LPS)-induced inflammatory injury of ATDC5 cells. METHODS: ATDC5 cells were cultured and treated with LPS in a series of concentration (0, 1, 5, and 10 µg/ml) for 5 h. The cells were transfected with miR-1246-mimic, inhibitor, si-HNF4γ or negative control, then were assessed for cell viability using CCK8 assay, apoptosis by flow-cytometry and expressions of miR-1246 and pro-inflammatory cytokines by qRT-PCR and western blot analysis. RESULTS: Cell viability was significantly reduced and cell apoptosis was added in ATDC5 cells injured with LPS at the dosage of 5 and 10 µg/ml. Relative mRNA expressions of pro-inflammatory cytokines (IL-1ß, IL-6, IL-8 and TNF-α) were significantly increased. miR-1246 was up-regulated in ATDC5 cells treated with LPS. Moreover, miR-1246 overexpression aggravated LPS-induced decrease in cell viability, increase in apoptosis and overproduction of pro-inflammatory factors. mRNA and protein expressions of HNF4γ were significantly suppressed in cells transfected with miR-124-mimic. Further, miR-1246 knockdown alleviated LPS-induced inflammatory injury by up-regulating the expression of HNF4γ and activation of PI3K/AKT and JAK/STAT pathways. CONCLUSIONS: Suppression of miR-1246 alleviated LPS-induced inflammatory injury in chondrogenic ADTC5 cells by up-regulation of HNF4γ and activation of PI3K/AKT and JAK/STAT pathways. The findings of this study will provide a novel viewpoint regarding miR-1246 target for clinical.

Long non-coding RNA HOXA11-AS functions as a competing endogenous RNA to regulate ROCK1 expression by sponging miR-124-3p in osteosarcoma.

Long non-coding RNAs (lncRNAs) have been strongly associated with various types of cancer. this study was to explore the critical role of lncRNA HOXA11-AS in osteosarcoma (OS) progression. Briefly, we should that the expression of HOXA11-AS was upregulated in OS tissues and cell lines. The high expression of HOXA11-AS was associated with advanced clinical stage, distant metastasis and poor overall survival of OS. In addition, We found that HOXA11-AS silencing suppressed OS cells proliferation, invasion and induced cell arrest in G0/G1 phase. Furthermore, our data showed that HOXA11-AS acts as an endogenous sponge by directly binding miR-124-3p, and decreasing the expression of miR-124-3p. Moreover, we found that HOXA11-AS may regulate tumor progression by affecting miR-124-3p targets, and ROCK1 expression. To conclude, our study helps to elucidate the effectiveness of HOXA11-AS promotion on OS cell proliferation and metastasis. A better understanding of interaction mechanism between HOXA11-AS-miR-124-3p-ROCK1 signaling axis may be a step forward in the development of new therapeutic strategies for the treatment of OS.

Salinity Stress Is Beneficial to the Accumulation of Chlorogenic Acids in Honeysuckle (Lonicera japonica Thunb.).

Honeysuckle (Lonicera japonica Thunb.) is a traditional medicinal plant in China that is particularly rich in chlorogenic acids, which are phenolic compounds with various medicinal properties. This study aimed to examine the effects of salinity stress on accumulation of chlorogenic acids in honeysuckle, through hydroponic experiments and field trials, and to examine the mechanisms underlying the effects. NaCl stress stimulated the transcription of genes encoding key enzymes in the synthesis of chlorogenic acids in leaves; accordingly, the concentrations of chlorogenic acids in leaves were significantly increased under NaCl stress, as was antioxidant activity. Specifically, the total concentration of leaf chlorogenic acids was increased by 145.74 and 50.34% after 30 days of 150 and 300 mM NaCl stress, respectively. Similarly, the concentrations of chlorogenic acids were higher in the leaves of plants in saline, compared with non-saline, plots, with increases in total concentrations of chlorogenic acids of 56.05 and 105.29% in October 2014 and 2015, respectively. Despite leaf biomass reduction, absolute amounts of chlorogenic acids per plant and phenylalanine ammonia-lyase (PAL) activity were significantly increased by soil salinity, confirming that the accumulation of chlorogenic acids in leaves was a result of stimulation of their synthesis under salinity stress. Soil salinity also led to elevated chlorogenic acid concentrations in honeysuckle flower buds, with significant increases in total chlorogenic acids concentration of 22.42 and 25.14% in May 2014 and 2015, respectively. Consistent with biomass reduction, the absolute amounts of chlorogenic acid per plant declined in flower buds of plants exposed to elevated soil salinity, with no significant change in PAL activity. Thus, salinity-induced chlorogenic acid accumulation in flower buds depended on an amplification effect of growth reduction. In conclusion, salinity stress improved the medicinal quality of honeysuckle by promoting accumulation of chlorogenic acids, however, the mechanisms underlying this process were not consistent in flower buds and leaves. Honeysuckle appears to be a promising plant for cultivation in saline land. Our study deepens knowledge of medicinal plant ecology and may provide a guide for developing saline agriculture.

Transcript levels of major interleukins in relation to the clinicopathological profile of patients with tuberculous intervertebral discs and healthy controls.

OBJECTIVES: The purpose of the present study was to simultaneously examine the transcript levels of a large number of interleukins (ILs; IL-9, IL-10, IL-12, IL-13, IL-16, IL-17, IL-18, IL-26, and IL-27) and investigate their correlation with the clinicopathological profiles of patients with tuberculous intervertebral discs. METHODS: Clinical data were collected from 150 patients participating in the study from January 2013 to December 2013. mRNA expression levels in 70 tuberculous, 70 herniated, and 10 control intervertebral disc specimens were determined by real-time polymerase chain reaction. RESULTS: IL-10, IL-16, IL-17, IL-18, and IL-27 displayed stronger expression in tuberculous spinal disc tissue than in normal intervertebral disc tissue (P<0.05). Our results illustrated multiple correlations among IL-10, IL-16, IL-17, IL-18, and IL-27 mRNA expression in tuberculous samples. Smoking habits were found to have a positive correlation with IL-17 transcript levels and a negative correlation with IL-10 transcript levels (P<0.05). Pain intensity, symptom duration, C-reactive protein levels, and the erythrocyte sedimentation rate exhibited multiple correlations with the transcript levels of several ILs (P<0.05). CONCLUSIONS: The experimental data imply a double-sided effect on the activity of ILs in tuberculous spinal intervertebral discs, suggesting that they may be involved in intervertebral discs destruction. Our findings also suggest that smoking may affect the intervertebral discs destruction process of spinal tuberculosis. However, further studies are necessary to elucidate the exact role of ILs in the intervertebral discs destruction process of spinal tuberculosis.

[Vertical variability of Pinus sylvestris var. mongolica tree ring delta13C and its relationship with tree ring width in northern Daxing' an Mountains of Northeast China].

A measurement was made on the vertical direction tree ring stable carbon isotope ratio (delta13C) and tree ring width of Pinus sylvestris var. mongolica in northern Daxing' an Mountains of Northeast China, with the relationship between the vertical direction variations of the tree ring delta13C and tree ring width analyzed. In the whole ring of xylem, earlywood (EW) and bark endodermis, the delta13C all exhibited an increasing trend from the top to the base at first, with the maximum at the bottom of tree crown, and then, decreased rapidly to the minimum downward. The EW and late-wood (LW) had an increasing ratio of average tree ring width from the base to the top. The average annual sequence of the delta13C in vertical direction had an obvious reverse correspondence with the average annual sequence of tree ring width, and had a trend comparatively in line with the average annual sequence of the tree ring width ratio of EW to LW above tree crown. The variance analysis showed that there existed significant differences in the sequences of tree ring delta13C and ring width in vertical direction, and the magnitude of vertical delta13C variability was basically the same as that of the inter-annual delta13C variability. The year-to-year variation trend of the vertical delta13C sequence was approximately identical. For each sample, the delta13C sequence at the same heights was negatively correlated with the ring width sequence, but the statistical significance differed with tree height.

[Establishment of a mouse model bearing spinal metastatic human prostate cancer using fluorescence-labeled PC-3 cells].

OBJECTIVE: To establish a mouse model of spinal metastasis of human prostate cancer using fluorescence-labeled PC-3 cells to allow direct observation by in vivo imaging. METHODS: PC-3 cells were infected with a lentivirus carrying green fluorescence protein (GFP) gene. The GFP-positive cell clone was expanded and prepared into cell suspension for injection into the inferior vena cava of nude mice. The tumor growth and metastasis in the mice was directly observed using an in vivo fluorescence imaging system. The tumor-bearing mice were sacrificed after 3 months for histological examination with HE staining. RESULTS: The labeled cells showed stable GFP expression both in vitro and in vivo. One week after cell injection, green fluorescence signals were detected by the in vivo fluorescence imaging system in the lower back of the mice, and at 4 weeks, the fluorescent tumor mass increased with a bone metastasis rate of 19% (3/16). Dissection of the mice at 3 months revealed lumbar tumor infiltration in 3 mice, showing a consistent result with in vivo fluorescence imaging. CONCLUSION: The nude mouse model of spinal bone metastasis of human prostate cancer established using GFP-labeled PC-3 cells facilitates further study of bone metastasis of prostate cancer.

[Responses of Pinus tabulaeformis forest ecosystem in North China to climate change and elevated CO2: a simulation based on BIOME-BGC model and tree-ring data].

Based on BIOME-BGC model and tree-ring data, a modeling study was conducted to estimate the dynamic changes of the net primary productivity (NPP) of Pinus tabulaeformis forest ecosystem in North China in 1952-2008, and explore the responses of the radial growth and NPP to regional climate warming as well as the dynamics of the NPP in the future climate change scenarios. The simulation results indicated the annual NPP of the P. tabulaeformis ecosystem in 1952-2008 fluctuated from 244.12 to 645.31 g C x m(-2) x a(-1), with a mean value of 418.6 g C x m(-2) x a(-1) The mean air temperature in May-June and the precipitation from previous August to current July were the main factors limiting the radial growth of P. tabulaeformis and the NPP of P. tabulaeformis ecosystem. In the study period, both the radial growth and the NPP presented a decreasing trend due to the regional warming and drying climate condition. In the future climate scenarios, the NPP would have positive responses to the increase of air temperature, precipitation, and their combination. The elevated CO2 would benefit the increase of the NPP, and the increment would be about 16.1% due to the CO2 fertilization. At both ecosystem and regional scales, the tree-ring data would be an ideal proxy to predict the ecosystem dynamic change, and could be used to validate and calibrate the process-based ecosystem models including BIOME-BGC.

[Responses of Pinus sylvestris var. mongolica radial growth to climate warming in Great Xing' an Mountins: a case study in Mangui].

Based on the theory and methodology of dendrochronology, the tree ring width chronology of Pinus sylvestris var. mongolica in Mangui of Great Xing' an Mountains was developed, and the relationships between the standardized tree ring width chronology and local climate factors (temperature and precipitation) as well as the effects of climate factors on the P. sylvestris var. mongolica radial growth were analyzed. In this region, the mean monthly temperature in April-August of current year was the main factor limiting the radial growth, and the increasing mean monthly temperature from April to August had negative effects to the radial growth. The simulation of the variations of the radial growth by the mean monthly temperature change in April-August showed that the radial growth of P. sylvestris var. mongolica would present a declining trend accompanied with the warmer and drier regional climate condition.

[Larix gmelinii tree-ring width chronology and its responses to climate change in Kuduer, Great Xing' an Mountains].

Based on the established tree-ring width chronology of Larix gmelinii in Kuduer, Great Xing' an Mountains, this paper analyzed the relationships between L. gmelinii tree-ring width chronology and related climatic variables, including air temperature, precipitation, and PDSI. In the study area, the L. gmelinii tree-ring width chronology was significantly negatively correlated with the air temperature in May and July (P<0.01), had no significant correlation with precipitation, but significantly positively correlated with the PDSI in June-August (P<0.05), suggesting that hydro-thermal variables had significant coupling effects to the radial growth of L. gmelinii in its growth season, particularly in May and July. The L. gmelinii tree-ring width chronology had significant correlations with large scale climatic regimes such as the Pacific Decadal Oscillation (PDO) in both low and high frequency variations, illustrating the significant effects of the remote oceanic climatic regimes to the local tree growth.

[Dendrochronology of Chinese pine in Mulan-Weichang, Hebei Province: a primary study].

Dendroclimatic methods were used to investigate the relationships between the growth of Chinese pine (Pinus tabulaeformis Carr.) and the climatic parameters in Mulan-Weichang of Hebei Province. The results showed that Chinese pine presented high sensitivity to climatic changes, and its earlywood width showed the highest sensitivity. There was a significant negative correlation between the tree-ring width chronology of Chinese pine and the air temperature in May-June. The precipitation and relative humidity in June had strong positive effects on the growth of earlywood, the precipitation from September to next September had significant positive effects on Chinese pine growth, and the relative humidity in winter more strongly affected the growth of latewood than of earlywood. There was a definite correlation between the tree-ring width chronology of Chinese pine and the large scale climate fluctuation. From 1951 to 2006, the increase of air temperature in study area was significant, and the sensitivity of Chinese pine to the variations of local temperature and precipitation decreased, presenting an inverse transforming trend with increasing temperature. Greater differences were observed between the reconstructed and observed data of mean temperature in May - June in a century scale, suggesting that the tree-ring growth of Chinese pine in study area had a greater fluctuation of sensitivity to the variation of climatic factors.