This study examined the potential biomedical properties of a bark extract from Pinus massoniana Lamb (PMBE). Its antioxidant activity, superoxide anion radical scavenging and free radical scavenging activities were examined. The antioxidant activity in a linoleic acid emulsion increased with increasing amounts of PMBE, with 100, 300 and 500 microg PMBE inhibiting 64.7%, 68.6% and 76.6% of peroxidation, respectively. Similarly, the radical scavenging activity by PMBE increased in a dose-dependent manner, suggesting that there may be a correlation between the antioxidant and scavenging activities. In addition, the effect of PMBE on human hepatocellular carcinoma BEL-7402 cells and normal liver L-02 cells was investigated in vitro. PMBE appeared to inhibit selectively the growth of BEL-7402 and slightly promoted the growth of L-02. Taken together, the results indicate that PMBE is a natural antioxidant that could potentially be used as a food supplement or as a candidate precursor substance for new anticancer therapeutics.
The ~24-amino-acid leucine-rich tandem repeat motif (PXXXXXLXXLXXLXLSXNXLXGXI) of carrot antifreeze protein comprises most of the processed protein and should contribute at least partly to the ice-binding site. Structural predictions using publicly available online sources indicated that the theoretical three-dimensional model of this plant protein includes a 10-loop β-helix containing the ~24-amino-acid tandem repeat. This theoretical model indicated that conservative asparagine residues create putative ice-binding sites with surface complementarity to the 1010 prism plane of ice. We used site-specific mutagenesis to test the importance of these residues, and observed a distinct loss of thermal hysteresis activity when conservative asparagines were replaced with valine or glutamine, whereas a large increase in thermal hysteresis was observed when phenylalanine or threonine residues were replaced with asparagine, putatively resulting in the formation of an ice-binding site. These results confirmed that the ice-binding site of carrot antifreeze protein consists of conservative asparagine residues in each β-loop. We also found that its thermal hysteresis activity is directly correlated with the length of its asparagine-rich binding site, and hence with the size of its ice-binding face.
Network science have constantly been in the focus of research for the last decade, with considerable advances in the controllability of their structural. However, much less effort has been devoted to study that how to improve the controllability of complex networks. In this paper, a new algorithm is proposed to improve the controllability of complex networks by rewiring links regularly which transforms the network structure. Then it is demonstrated that our algorithm is very effective after numerical simulation experiment on typical network models (Erd\"os-R\'enyi and scale-free network). We find that our algorithm is mainly determined by the average degree and positive correlation of in-degree and out-degree of network and it has nothing to do with the network size. Furthermore, we analyze and discuss the correlation between controllability of complex networks and degree distribution index: power-law exponent and heterogeneity
In chloroplasts, thioredoxin (TRX) isoforms and NADPH-dependent thioredoxin reductase C (NTRC) act as redox regulatory factors involved in multiple plastid biogenesis and metabolic processes. To date, less is known about the functional coordination between TRXs and NTRC in chlorophyll biosynthesis. In this study, we aimed to explore the potential functions of TRX m and NTRC in the regulation of the tetrapyrrole biosynthesis (TBS) pathway. Silencing of three genes, TRX m1, TRX m2, and TRX m4 (TRX ms), led to pale-green leaves, a significantly reduced 5-aminolevulinic acid (ALA)-synthesizing capacity, and reduced accumulation of chlorophyll and its metabolic intermediates in Arabidopsis (Arabidopsis thaliana). The contents of ALA dehydratase, protoporphyrinogen IX oxidase, the I subunit of Mg-chelatase, Mg-protoporphyrin IX methyltransferase (CHLM), and NADPH-protochlorophyllide oxidoreductase were decreased in triple TRX m-silenced seedlings compared with the wild type, although the transcript levels of the corresponding genes were not altered significantly. Protein-protein interaction analyses revealed a physical interaction between the TRX m isoforms and CHLM. 4-Acetoamido-4-maleimidylstilbene-2,2-disulfonate labeling showed the regulatory impact of TRX ms on the CHLM redox status. Since CHLM also is regulated by NTRC (Richter et al., 2013), we assessed the concurrent functions of TRX m and NTRC in the control of CHLM. Combined deficiencies of three TRX m isoforms and NTRC led to a cumulative decrease in leaf pigmentation, TBS intermediate contents, ALA synthesis rate, and CHLM activity. We discuss the coordinated roles of TRX m and NTRC in the redox control of CHLM stability with its corollary activity in the TBS pathway.
To observe clinical effects of acupressure combined with activating blood and dissipating blood stasis prescription for the treatment of early swelling of calcaneal fractures.From June 2016 to April 2018, 78 patients with calcaneus were divided into observation group and control group. There were 39 patients in observation group, including 36 males and 3 females aged from 28 to 61 years old with an average of(41.7±13.3) years old; 20 patients were caused by falling down, 10 patients were caused by traffic accident and 9 patients were caused by other injuries; The time from injury to medical treatment ranged from 0.5 to 10.5 h with an average of(4.6±3.2) h; 15 patients were type II, 17 patients were type III and 7 patients were type IV according to Sanders classification; the patients began to be pressed the three points of "Xuehai(SP 10)" "Sanyinjiao(SP 6)" "Zusanli(ST 36)" for 10 min from the day of hospital admission to the day of operation, 3 times a day, and taken prescription of activating blood and dissipating blood stasis prescription orally for 5 days, one dose per day and twice a day. There were 39 patients in control group, including 34 males and 5 females aged from 26 to 62 years old with an average of (43.3±12.3) years old; 19 patients were caused by falling down, 12 patients were caused by traffic accident and 8 patients were caused by other injuries; the time from injury to medical treatment ranged from 1 to 11 h with an average of (4.4±3.7) h; 15 patients were typeII, 18 patients were type III and 6 patients were type IV according to Sanders classification; patients were given 20% mannitol intravenously for 5 days, twice a day, 125 ml each time from hospital admission. During the 5 days of admission, the degree of swelling of the affected ankle was measured and calculated by water spillover method every day. The result which was measured and calculated on the first day of admission without treatment was the degree of swelling before treatment, compared degree of swelling before operation and 1st, 2nd, 3rd and 4th day after treatment; the time needed for the first positive skin fold sign in the affected ankle and the number of cases of tension blisters between two groups before operation were observed.The swelling of the affected ankle on the1st, 2nd, 3rd and 4th day after treatment in the observation group separately were(12.67±0.82)%, (11.87±0.88)%, (10.65±0.92)%, (9.47±0.96)%, and lower than control group which separately were(13.31±0.98)%, (13.51±0.84)%, (12.22±0.94)%, (11.38±1.01)%; The time for the first appearance of the skin fold sign was (6.41±1.74) days in the observation group was earlier than that of the control group (8.15±2.01) days; There was no significant difference in occurrence of tension blisters between observation group(3 patients) and control group(9 patients).Acupressure combined with activating blood and dissipating blood stasis prescription for the treatment of early swelling of calcaneal fractures could accelerate the decline of swelling of the affected ankle, shorten the time of the first skin fold sign, reduce the incidence of tension blisters in the ankle, shorten the waiting time of operation, and the surgical treatment as soon as possible.
Abstract Background Plant protoplasts, a proven physiological and versatile cell system, are widely used in high-throughput analysis and functional characterization of genes. Green protoplasts have been successfully used in investigations of plant signal transduction pathways related to hormones, metabolites and environmental challenges. In rice, protoplasts are commonly prepared from suspension cultured cells or etiolated seedlings, but only a few studies have explored the use of protoplasts from rice green tissue. Results Here, we report a simplified method for isolating protoplasts from normally cultivated young rice green tissue without the need for unnecessary chemicals and a vacuum device. Transfections of the generated protoplasts with plasmids of a wide range of sizes (4.5-13 kb) and co-transfections with multiple plasmids achieved impressively high efficiencies and allowed evaluations by 1) protein immunoblotting analysis, 2) subcellular localization assays, and 3) protein-protein interaction analysis by bimolecular fluorescence complementation (BiFC) and firefly luciferase complementation (FLC). Importantly, the rice green tissue protoplasts were photosynthetically active and sensitive to the retrograde plastid signaling inducer norflurazon (NF). Transient expression of the GFP-tagged light-related transcription factor OsGLK1 markedly upregulated transcript levels of the endogeneous photosynthetic genes OsLhcb1 , OsLhcp , GADPH and RbcS , which were reduced to some extent by NF treatment in the rice green tissue protoplasts. Conclusions We show here a simplified and highly efficient transient gene expression system using photosynthetically active rice green tissue protoplasts and its broad applications in protein immunoblot, localization and protein-protein interaction assays. These rice green tissue protoplasts will be particularly useful in studies of light/chloroplast-related processes.
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