miR-940 promotes spinal cord injury recovery by inhibiting TLR4/NF-κB pathway-mediated inflammation.

OBJECTIVE: The aim of this study was to investigate the effects of miR-940 and Toll-like receptor 4/Nuclear Factor κB (TLR4/NF-κB) pathways on inflammatory responses and spinal cord injury (SCI). MATERIALS AND METHODS: This study first established a model of spinal cord injury in mice. The grip force measurement was used to detect the recovery of the forelimb, left forelimb and right forelimb of SCI mice. The quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was used to detect the expression of miR-940 and macrophage receptor TLR4 in SCI mice. In addition, the protein levels of TLR4 and inducible nitric oxide synthase (iNOS) in SCI mice were detected by Western blot. MiR-940 mimic was injected into the injured area of SCI mice to explore the effect of miR-940 overexpression on TLR4 and myeloperoxidase (MPO) expression as well as the protein levels of TLR4, P65 and iNOS. Furthermore, the grip strength of SCI mice with double forelimb, left forelimb and right forelimb was detected by the grip force test after miR-940 overexpression. RESULTS: Compared with the sham-operated mice, the grip strength of the forelimb, left forelimb, and right forelimb of the SCI group showed significant obstacles. Meanwhile, the expression of miR-940 was remarkably decreased in SCI mice along with significant elevation of the inflammatory response-related factors including TRL4 and iNOS. Then we injected SCI mice with miR-940 mimics into the spinal cord injury area and found that miR-940 overexpression decreased the expression levels of TLR4 and MPO. At the same time, the overexpression of miR-940 markedly decreased the protein levels of TLR4, P65, and iNOS in SCI mice. In addition, miR-940 overexpression improved the grip strength of the left and right forepaws and the simultaneous grip strength of the two claws of the SCI mice than those of the simple injury group. CONCLUSIONS: High expression of miR-940 can promote the recovery of spinal cord injury by downregulating the TLR4/NF-κB signaling pathway and inhibiting inflammation.

Optical determination of spin diffusion length and interfacial spin mixing conductance in epitaxial Pd/Fe bilayers.

We present a detailed study of the photoinduced magnetization precession and magnetic relaxation in epitaxial Pd/Fe films on MgO(0 0 1) substrates via all-optical pump-probe techniques. We explicitly formulate the correlation between the Gilbert damping and the effective damping in the optical approach. Furthermore, a non-local Gilbert damping induced by spin pumping is demonstrated self-consistently by the Gilbert damping dependence on Pd thickness and the [Formula: see text] relationship of damping with Fe thickness [Formula: see text]. The non-local Gilbert damping enables the determination of spin diffusion length [Formula: see text] in Pd and interfacial spin mixing conductance [Formula: see text]. Our work paves the way toward the optical determination of spin transport parameters.

MicroRNA-23c inhibits articular cartilage damage recovery by regulating MSCs differentiation to chondrocytes via reducing FGF2.

OBJECTIVE: The aim of the study was to explore the role of microRNA-23c in the differentiation of marrow stromal cells (MSCs) to chondrocytes and its potential mechanism. MATERIALS AND METHODS: MSCs were first isolated from rat bone marrow for cell culture. Surface antigens of MSCs (CD29 and CD34) were identified by flow cytometry. MSCs were induced for chondrogenic differentiation in MCDM (Mesenchymal Stem Cell Chondrogenic Differentiation Medium) for 0, 3, and 7 days, respectively, followed by detection of RUNX2, microRNA-23c and FGF2 expressions by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Alcian blue staining was performed to access proteoglycan deposition in MSCs transfected with microRNA-23c mimics or inhibitor. Western blot was conducted to detect the protein expressions of ACAN and COL2A1 in MSCs. The binding condition between microRNA-23c and FGF2 was verified by dual-luciferase reporter gene assay. Finally, MSCs were co-transfected with microRNA-23c mimics and FGF2 overexpression plasmid for rescue experiments. RESULTS: On the fourth day of MSCs isolation, MSCs were in an elongated shape. Flow cytometry results showed positive expression of CD29 and negative expression of CD34, which were consistent with MSCs phenotype. QRT-PCR data elucidated that the mRNA levels of RUNX2 and FGF2 gradually increased, whereas microRNA-23c expression decreased with the prolongation of chondrogenic differentiation. Transfection of microRNA-23c mimics in MSCs remarkably elevated microRNA-23c expression. Alcian blue staining showed that microRNA-23c overexpression results in less proteoglycan deposition in MSCs than that of controls. Both mRNA and protein expressions of ACAN and COL2A1 decreased after microRNA-23c overexpression. Dual-luciferase reporter gene assay confirmed that FGF2 binds to microRNA-23c. Further Western blot results demonstrated that FGF2 expression is negatively regulated by microRNA-23c. FGF2 overexpression reversed the inhibitory effects of microRNA-23c on proteoglycan deposition, as well as expressions of ACAN and COL2A1. CONCLUSIONS: MicroRNA-23c expression decreases during chondrogenic differentiation of MSCs, which inhibits MSCs differentiation to chondrocytes by inhibiting FGF2.

[miR-30a-5p promotes the apoptosis of chondrocytes in patients with osteoarthritis by targeting protein kinase B].

Objective: To investigate the expression of miR-30a-5p in cartilage of osteoarthritis patients, and to explore its mechanism of chondrocyte apoptosis. Methods: From May 2015 to December 2016, tissue specimen of 289 patients with osteoarthritis was collected in Department of Orthopedics, Changzhou traditional Chinese Medicine Hospital Affiliated to Nanjing University of Chinese Medicine.The expression of miR-30a-5p and protein kinase B(Akt) mRNA in cartilage of different patients was detected by qPCR.The apoptosis of chondrocytes was detected by Tunel method.The expression of related proteins in tissues and cells was detected by immunoblotting, and apoptosis and cell cycle were detected by flow cytometry. Results: The expression of miR-30a-5p in OA patients was significantly higher than control patients (P<0.05), but Aktwas positively related[(3.64±0.95)vs(1.03±0.31), P<0.05]. The expression of miR-30a-5p in cartilage of OA patients was negatively correlated with Akt mRNA expression (r=0.729 3, P<0.001), but it had a positive correlation with the apoptotic rate (r=0.847 5, P<0.001). miR-30a-5p targets negative regulation of Akt gene expression in SW1353 cells, and the expression of p-Akt, IkB-α, p-IkB-α, p65, p-p65 and mTOR and p-mTOR were significantly down-regulated by miR-30a-5p (P<0.05). Compared with normal SW1353 cells, the apoptosis rate of SW1353 cells which was transfected with miR-30a-5p-mimics increased by 9.65 times, G0/G1 phase cells increased by 1.37 times, S phase cells decreased by 60.94%, G2/M phase cells decreased 19.53%. Conclusion: miR-30a-5p is highly expressed in cartilage of osteoarthritis patients, and its high expression can block chondrocytes in G0/G1 phase by targeting Akt gene, and induce apoptosis of chondrocytes.

First Report of Fusarium tricinctum Causing Stem and Root Rot on Lanzhou Lily (Lilium davidii var. unicolor) in China.

Lanzhou lily (Lilium davidii var. unicolor Cotton) is an important bulb edible crop which mostly distributes in middle area of Gansu Province in China (2). Recently, plants of Lanzhou lily developed symptoms of severe wilting. In early autumn of 2012 to 2013, a survey of Lanzhou lily disease was carried out in Yuanjiawan, Caoyuan, Xiguoyuan, and Hutan villages of Lanzhou City and Xuding and Guanshan villages of Linxia Prefecture. Disease symptoms included stem and root rot, vessels showed a brown to dark brown discoloration, plus a progressive yellowing and wilting of leaves from the base. Small pieces of symptomatic leaves, stems, and roots were surface disinfected with 75% ethanol for 30 s, 3% sodium hypochlorite for 5 min, and then washed three times in sterile distilled water. The tissues were placed on Martin Agar at 25°C for 7 days. Three isolates were consistently isolated from diseased tissues and all isolates with morphology similar to Fusarium spp. Isolates were transferred to potato dextrose agar (PDA) and carnation leaf agar (CLA) and incubated at 25°C in darkness. These isolates grew rapidly on PDA and formed abundant dense aerial mycelium, initially white, that became deep pink with age and formed red pigments in the medium. On CLA, macroconidia with 3 to 5 septa were abundant, relatively slender, and curved to lunate. Microconidia were abundant, oval and 0 to 1 septa. Chlamydospores were globose with a smooth outer wall in chains. The rDNA internal transcribed spacer (ITS) region comprising ITS1, ITS2, and 5.8S rDNA was amplified using primers ITS-1 and ITS-4 (3) and sequenced. On the basis of a comparison of 563 bp, all the three isolates had the identical sequence (GenBank Accession No. KF728675). BLASTn analysis of the sequence showed 100% match with the ITS sequences of those F. tricinctum sequences in GenBank (Accession Nos. FJ233196, AY188923, and JF776663). Pathogenicity test was performed by transplanting 2-month-old tissue culture seedlings to plastic pots in a sterile mixture of vermiculite and torf substrate at 1:3 (v/v). Seedlings were inoculated with 6 ml of the conidial suspension (104 conidia/ml) on the roots of plant in each pot, three plants per pot, and three replicates for each treatment. Seedlings treated with sterile water served as controls. The seedlings were placed in a plant growth chamber maintained at 22 ± 3°C, relative humidity >70%, 16 h light per day, and irrigated with sterile water. After 4 weeks, inoculated plants exhibited wilting foliage that with symptoms similar to those observed in the field, while the control plants remained healthy. F. tricinctum was re-isolated from all inoculated plants. The disease has been reported previously in ornamental lily in China (1). However, to the best of our knowledge, this is the first report of F. tricinctum causing wilt on edible Lanzhou lily in China and the disease must be taken into consideration of current disease management. This work supported by NSFC No. 31370447 and Hundred Talents Program of CAS "Molecular mechanism of biological control on plant diseases." References: (1) Y. Y. Li et al. Plant Dis. 97:993, 2013. (2) R. Y. Wang et al. Virol. J. 7:34, 2010. (3) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.

First Report of Lily Blight and Wilt Caused by Fusarium tricinctum in China.

Lily (Lilium spp.) is one of the most well-known horticultural crops, and plays an important economic role in China. In September 2011, wilted plants were observed on Lilium oriental hybrid cultivar 'Sorbonne' growing in Longde County, Ningxia Hui Autonomous Region, China. Disease symptoms included wilting, stem and root rot, brown spots of bulbs and then bulbs rotting and spalling from the basal disc, plus a progressive yellowing and defoliation of the leaves from the base. Diseased plants were sampled from fields. Small pieces of symptomatic bulbs, stems, and roots from 10 different plants were surface disinfected with 75% ethanol for 30 s, 3% sodium hypochlorite for 5 min, and then washed three times in sterilize distilled water. The tissues were placed onto Martin Agar (2) at 25°C for 7 days. Nine isolates with morphology similar to Fusarium were obtained from the diseased tissues. Isolates were transferred to potato dextrose agar (PDA) and carnation leaf agar (CLA) and incubated at 25°C. Seven were identified as Fusarium oxysporum and one was F. solani, which have been reported as pathogens of lily in China (1). The other isolate, when grown on PDA, rapidly produced dense, white aerial mycelium that became pink with age and formed red pigments in the medium. On CLA, macroconidia with three to five septate were abundant, relatively slender, and curved to lunate. Microconidia were abundant, oval or pyriform, and one to two celled. Chlamydospores were in chains with smooth exine. The rDNA internal transcribed spacer (ITS) region and a portion of the translation elongation factor 1-alpha (EF-1α) gene of the fungus were amplified, with universal primers ITS1/ITS4 and EF1/EF2 primers respectively (3) and sequenced. In addition, the ß-tubulin gene (ß-tub) of the fungus was amplified with modified primers Btu-F-F01 (5'-CAGACMGGTCAGTGCGTAA-3') and Btu-F-R01 (5'-TCTTGGGGTCGAACATCTG-3') (4). BLASTn analysis showed that the ITS sequences of the isolate (GenBank Accession No. JX989827) had 98.9% similarity with those of F. tricinctum (EF611092, JF776665, and HM776425) and the EF-1α sequences of the isolate (JX989828) had 98.1% similarity with those of F. tricinctum (EU744837 and JX397850). The ß-tub sequences of the isolate (JX989829) had 99.0% similarity with those of F. tricinctum (EU490236 and AB587077). The isolate was tested for pathogenicity. Two-month-old 'Sorbonne' seedlings were inoculated by placing 5 ml of conidial suspension (about 106 conidia per ml) over the roots of plants in each pot. Control plants were treated with sterile water in the same way. Plants were placed in a greenhouse at 22 to 25°C with a 15-h photoperiod. There were eight plants per pot and three replicates for each treatment. After 3 weeks, 87.5% of the inoculated plants exhibited browning of the root tips, root rot, and yellowing of the leaves, while control plants were symptomless. The pathogen was reisolated from the infected roots and identified as F. tricinctum, thus fulfilling Koch's postulates. To our knowledge, this is the first report of Fusarium wilt of lily caused by F. tricinctum. This information will provide guidance for the control of lily wilt disease and add information useful for the production of lilies. References: (1) C. Li and J. J. Li. Acta Phytopathol. Sin. (in Chinese) 26:192, 1995. (2) J. P. Martin. Soil Sci. 38:215, 1950. (3) K. O'Donnell et al. Proc. Nat. Acad. Sci. U. S. A. 95:2044, 1998. (4) M. Watanabe et al. BMC Evol. Biol. 11:322. 2011.

[Isolation and fermentation conditions of polyvinyl alcohol-degrading enzyme producing strain].

A bacterium D8 strain which high efficiently degrading PVA was isolated from waste water of factory. The strain possesses the abilities of completely degrading 0.5 per cent of PVA (500, 1700) included in the culture medium for four days. It was identified Pseudomonas pseudoalcaligenes. Fermentation conditions of the strain have been investigated. The suitable medium consisted of PVA 1.5% (NH4)2SO40.1%, K2HPO4 0.24%, KH2PO4 0.04%, MgSO4.7H2O 0.035%, NaCl 0.01%, FeSO4 0.001%, yeast extract 0.15%, pH 7.5. The optimal condition for enzyme production are as follows: 250 ml shake filled with 30 ml medium, 30 degrees C, 160n/min incubation period 72 h. Under such conditions enzyme activity is highest.