Electrohydrodynamic drying of citrus (Citrus sinensis L.) peel: Comparative evaluation on the physiochemical quality and volatile profiles.

Based on the concept of circular economy, citrus peel was considered a valuable source of bioactive compounds for high-value foods. Electrohydrodynamic (EHD) drying is a novel technology appropriated for the dehydration of heat-sensitive products such as citrus peel. In current work, EHD drying of citrus peel was performed based on alternating current (AC) or direct current (DC) sources at various voltage levels (9, 18, 27, 36, and 45 kV). The effect of EHD on drying characteristics, water retention capacity, enzyme inactivation, phytochemical contents (phenolic compounds and carotenoids), and volatile compounds of citrus peel were evaluated and compared. Results showed that the drying time in the AC electric field was shorter compared to DC electric field at the same applied voltages due to the polarization layer formed by unipolar charges. The applied voltage determined electric field strength as well as the degree of tissue collapse and cell membrane rupture. EHD elucidated the transformation and degradation of phytochemicals including phenolic compounds, carotenoids, and volatile composition in proportion to the applied voltage. The findings indicate that EHD drying with AC improves drying behaviors, inactivates enzymes, and retains the phytochemical properties of citrus peel.

[Availability Changes in Different Exogenous Selenium Fertilizers in Soil and Their Effects on Selenium Accumulation in Wheat].

In order to understand the differences in the uptake and accumulation of several common exogenous selenium fertilizers by crops, a wheat pot experiment was conducted to study the availability changes in different selenium fertilizers (potassium selenate, potassium selenite, EDTA-chelated selenium, selenium powder, fly ash, and selenium-enriched straw) in soil and their effects on wheat growth and selenium uptake and distribution. The results showed that the change in availability of different exogenous selenium types in soil was different. During the whole growth period of wheat, the soil available selenium proportion of selenate, selenite, and EDTA-chelated selenium treatment was significantly higher than that of the control (CK), respectively, but there was no significant difference between the other treatments and the CK treatment. In the early stage of wheat growth, the soil available selenium proportion of selenate, selenite, and selenium powder treatment decreased gradually and tended to be stable in the later growth stage of wheat; however, the soil available selenium proportion of other exogenous selenium treatments showed a dynamic change of decreasing in the early period and increasing in the late period. The available selenium content in soil significantly affected the selenium uptake by wheat, and there was a significant positive correlation between them. Selenate application significantly increased the grain and leaf biomass of wheat, but other selenium fertilizers had no significant effect on wheat growth. The accumulation capacity of different exogenous selenium fertilizers for wheat followed the order of selenate>selenite, EDTA-chelated selenium>selenium powder, fly ash, and selenium-enriched straw. There was no significant difference between the selenium powder, fly ash, and selenium-enriched straw treatments and the CK treatment. Selenium was more easily transferred to and accumulated in the stems and leaves of wheat after the application of selenate, whereas selenium was more easily transferred to and accumulated in grains after the application of selenite and EDTA-chelated selenium.

Ameliorative effect of silymarin on the quality of frozen-thawed boar spermatozoa.

Although Silymarin (SMN) has powerful antioxidant properties, little is known about its effects on the quality of frozen-thawed boar sperm. The present study aimed to evaluate the influences of SMN added to the thawing extender on boar sperm parameters essential for fertilization. The frozen-thawed semen was diluted in a Modena thawing extender supplemented with different concentrations of SMN (0, 5, 10, 20 and 50 µM respectively), and then the changes in quality parameters, antioxidant capacity, mitochondrial function and in vitro fertilization (IVF) capability of frozen-thawed sperm were assessed. Here we demonstrated that the motility, plasma membrane integrity and acrosomal integrity of frozen-thawed sperm improved efficiently by SMN (p < .05). In antioxidant parameters evaluation, the tROS level and MDA content of frozen-thawed spermatozoa were reduced in the 20 µM SMN group, while the T-AOC activity significantly increased (p < .05), indicating that the supplementation with SMN can promote the antioxidant capacity of frozen-thawed boar sperm. Besides, we also discovered that the addition of SMN significantly upregulated ATP content and enhanced the mitochondrial activity of sperm. More interestingly, SMN promoted the activities of mitochondrial respiratory chain complexes (MRCC) I, II, III and IV in frozen-thawed sperm significantly. Functionally, the higher penetration rate and increased total efficiency of fertilization were observed in the 20 µM SMN group. In summary, supplementation with SMN in the thawing medium ameliorates the quality of frozen-thawed boar sperm by enhancing mitochondrial respiratory capacity, producing large amounts of ATP and regulating ROS formation.

Gamma Knife Radiosurgery for Cavernous Malformations of Basal Ganglia and Thalamus: A Retrospective Study of 53 Patients.

INTRODUCTION: Gamma Knife radiosurgery (GKRS) has been used to treat cavernous malformations (CMs) located in basal ganglia and thalamus. However, previous reports are limited by small patient population. METHODS: We retrospectively reviewed the clinical and radiological data of 53 patients with CMs of basal ganglia and thalamus who underwent GKRS at West China Hospital between May 2009 and July 2018. All patients suffered at least once bleeding before GKRS. The mean volume of these lesions was 1.77 cm3, and the mean marginal dose was 13.2 Gy. After treatment, patients were followed to determine the change in symptom and hemorrhage event. RESULTS: The mean follow-up period was 52.1 months (6.2-104.3 months). The calculated annual hemorrhage rate (AHR) was 48.5% prior to GKRS and 3.0% after treatment (p < 0.001). The Kaplan-Meier analysis revealed that 2-, 3-, and 5-year hemorrhage-free survival were 88, 80.9, and 80.9%, respectively. Preexisting symptoms were resolved in 11 patients, improved in 14, and stable in 5. Only 2 patients (3.8%) developed new neurological deficit. CONCLUSION: Our study suggests that AHR after GKRS was comparable to the recorded AHR of natural history (3.1-4.1%) in previous studies. GKRS is a safe and effective treatment modality for CMs of basal ganglia and thalamus. Considering the relative insufficient understanding of natural history of CMs, future study warrants longer follow-up.

Genome-wide identification and characterization of ALTERNATIVE OXIDASE genes and their response under abiotic stresses in Camellia sinensis (L.) O. Kuntze.

MAIN CONCLUSION: Four typical ALTERNATIVE OXIDASE genes have been identified in tea plants, and their sequence features and gene expression profiles have provided useful information for further studies on function and regulation. Alternative oxidase (AOX) is a terminal oxidase located in the respiratory electron transport chain. AOX catalyzes the oxidation of quinol and the reduction of oxygen into water. In this study, a genome-wide search and subsequent DNA cloning were performed to identify and characterize AOX genes in tea plant (Camellia sinensis (L.) O. Kuntze cv. Longjing43). Our results showed that tea plant possesses four AOX genes, i.e., CsAOX1a, CsAOX1d, CsAOX2a and CsAOX2b. Gene structure and protein sequence analyses revealed that all CsAOXs share a four-exon/three-intron structure with highly conserved regions and amino acid residues, which are necessary for AOX secondary structures, catalytic activities and post-translational regulations. All CsAOX were shown to localize in mitochondria using the green fluorescent protein (GFP)-targeting assay. Both CsAOX1a and CsAOX1d were induced by cold, salt and drought stresses, and with different expression patterns in young and mature leaves. Reactive oxygen species (ROS) accumulated strongly after 72 and 96 h cold treatments in both young and mature leaves, while the polyphenol and total catechin decreased significantly only in mature leaves. In comparison to AtAOX1a in Arabidopsis thaliana, CsAOX1a lost almost all of the stress-responsive cis-acting regulatory elements in its promoter region (1500 bp upstream), but possesses a flavonoid biosynthesis-related MBSII cis-acting regulatory element. These results suggest a link between CsAOX1a function and the metabolism of some secondary metabolites in tea plant. Our studies provide a basis for the further elucidation of the biological function and regulation of the AOX pathway in tea plants.

Transcriptome sequencing dissection of the mechanisms underlying differential cold sensitivity in young and mature leaves of the tea plant (Camellia sinensis).

The tea plant originated in tropical and subtropical regions and experiences considerable challenges during cold winters and late spring frosts. After short-term chilling stress, young leaves of tea plants exhibit browning, a significant increase in electrolyte leakage and a marked decrease in the maximal photochemical efficiency of photosystem II (Fv/Fm) compared with mature leaves. To identify the mechanisms underlying the different chilling tolerance between young and mature leaves of the tea plant, we used Illumina RNA-Seq technology to analyse the transcript expression profiles of young and mature leaves exposed to temperatures of 20 °C, 4 °C, and 0 °C for 4 h. A total of 45.70-72.93 million RNA-Seq raw reads were obtained and then de novo assembled into 228,864 unigenes with an average length of 601 bp and an N50 of 867 bp. In addition, the differentially expressed unigenes were identified via Venn diagram analyses for paired comparisons of young and mature leaves. Functional classifications based on Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses revealed that the up-regulated differentially expressed genes were predominantly related to the cellular component terms of chloroplasts and cell membranes, the biological process term of oxidation-reduction process as well as the pathway terms of glutathione metabolism and photosynthesis, suggesting that these components and pathways may contribute to the cold hardiness of mature leaves. Conversely, the inhibited expression of genes related to cell membranes, carotenoid metabolism, photosynthesis, and ROS detoxification in young leaves under cold conditions might lead to the disintegration of cell membranes and oxidative damage to the photosynthetic apparatus. Further quantitative real-time PCR testing validated the reliability of our RNA-Seq results. This work provides valuable information for understanding the mechanisms underlying the cold susceptibility of young tea plant leaves and for breeding tea cultivars with superior frost resistance via the genetic manipulation of antioxidant enzymes.

[Traditional Chinese Medicine data management policy in big data environment].

As traditional data management model cannot effectively manage the massive data in traditional Chinese medicine(TCM) due to the uncertainty of data object attributes as well as the diversity and abstraction of data representation, a management strategy for TCM data based on big data technology is proposed. Based on true characteristics of TCM data, this strategy could solve the problems of the uncertainty of data object attributes in TCM information and the non-uniformity of the data representation by using modeless properties of stored objects in big data technology. Hybrid indexing mode was also used to solve the conflicts brought by different storage modes in indexing process, with powerful capabilities in query processing of massive data through efficient parallel MapReduce process. The theoretical analysis provided the management framework and its key technology, while its performance was tested on Hadoop by using several common traditional Chinese medicines and prescriptions from practical TCM data source. Result showed that this strategy can effectively solve the storage problem of TCM information, with good performance in query efficiency, completeness and robustness.

Sesquiterpene amino ether and cytotoxic phenols from Dendrobium wardianum Warner.

A new bibenzyl derivative, dendrocandin V (1) and a new sesquiterpene amino ether, wardianumine A (2), together with eleven known compounds, including phenanthrenes (denbinobin (3), 9,10-dihydro-denbinobin (4), mostatin (5), loddigesiinols A (6)), bibenzyls (moscatilin (7), 5-hydroxy-3,4'-dimethoxybibenzyl (8), 3,4-dihydroxy-5,4'-dimethoxy bibenzyl (9), dendrocandin A (10), gigantol (11), dendrocandin U (12)) and an alkaloids (dihydroshihunine, 13) were isolated from the EtOH extraction of stems of Dendrobium wardianum Warner. Isolation of the new compound 2 indicated that N,N-dimethylethanolamine as the key adduction in the synthesis of dendroxine and its analogs in Dendrobium species. The hypothetical biosynthetic pathway of 2 was then postulated. Inspired by literature and traditional usage of the herbal medicine, some compounds were sent for cytotoxic activity and the results indicated that compounds 1, 3, 4, 5 showed cytotoxic activities against five human cancer cell lines (HL-60, A-549, SMMC-7721, MCF-7, and SW-480) with IC50 from 2.33-38.48µM. Among those compounds, 3 and 4 showed cell line selectivity with strong activity comparable to DDP.

Screening for differentially expressed genes in endophytic fungus strain 39 during co-culture with herbal extract of its host Dioscorea nipponica Makino.

Strain 39 is an endophytic fungus which was isolated from Dioscorea nipponica Makino (DNM). After Strain 39 co-cultured with ethanol extract of DNM rhizomes for several days, the content of saponins in this culture mixture would be obviously increased. To analyze the mechanism of this microbial transformation, we used the differential display reverse transcription polymerase chain reaction (DDRT-PCR) method to compare the transcriptomes between Strain 39 cultured in normal PD medium and in PD medium which added ethanol extract of DNM rhizomes. We amplified 29 DDRT-PCR bands using 12 primer combinations of three anchored primers and five random primers, and six bands were re-amplified. Analysis of real-time PCR and sequence alignment showed that three clones were up-regulated in sample group: squalene epoxidase, squalene synthase, and catalase, one clone was expressed only in sample group. The possible roles and origins of the above genes were discussed, and the molecular mechanism of Strain 39 biotransformation was speculated. This study is the first report of the molecular biotransformation mechanism of saponins production by endophytic fungus of DNM.

[Rapid identification of Dendrobium plants based on near infrared diffuse reflection spectroscopy].

Near infrared diffuse reflection spectra of 15 species' 171 samples of Dendrobium combined with chemometrics statistical analysis were used to build prediction model, in order to discriminate different species of Dendrobium quickly and nondestructively. Hotelling T2 was applied to stability analysis of spectrum of 5 random drawing samples, and the results showed that the samples spectrum possessed good stability. Orthogonal test L24 (2 x 4 x 3 x 8) was designed to optimize optical path type, spectral band, derivative and smooth. The result of orthogonal test was analyzed by principal component analysis, which revealed that when 6500-4000 cm(-1) spectral band was applied, and with multiplicative scatter correction, second derivative, Norris smooth, and the number of principal components 7, the spectrum distinguishing accuracy was 100%. With the optimized condition of orthogonal test as the input value of partial least squares discriminant analysis and random drawing 123 samples as calibration set to establish the prediction model, and the rest 48 samples as prediction set were use to assess the property of the prediction model, the results indicated that the accumulating contribution rate of the first 3 principal components of the model was 99.36%, the identification of the standard deviation was +/- 0.1, and the correct recognition rate of the model was 97.92%. The results were satisfied. The method provided a new way for the rapid identification of different species of Dendrobium, and also supplied a reference for the authentication of medicinal plants.

Reconstruct large osteochondral defects of the knee with hIGF-1 gene enhanced Mosaicplasty.

OBJECTIVE: To investigate a compound technique including gene therapy, injectable tissue engineering and Mosaicplasty to reconstruct large osteochondral defect. METHODS: Plasmid vector containing hIGF-1 cDNA was created and transfected into BMSCs in vitro with FuGene6. After gene expression determination, cells were mixed with calcium alginate gel. Osteochondral defects were created on the femoral condyle of goats in a diameter of 6mm. Osteochondral plugs were harvested from the intertrochlea groove and pressed into the recipient sites in a mosaic mode. Gene modified BMSCs-scaffold complex was applied to fill the residual defects. Control groups were also set up. At 4 and 16 weeks, specimens were investigated in gross and under microscopy, electromicroscopy and MRI detection. RESULTS: hIGF-I gene was expressed effectively with the peak concentration at 34.75 ng/ml. Subchondral bone and cartilage were integrated well in gene enhanced Mosaicplasty group. The reconstructed tissue filled up the gaps between columns, which appeared better than other groups. The regenerated cartilage was integrated with neighbor tightly in regular arrange. Extracellular matrix distributed evenly and deeply stained by alcian blue. Quantitative histologic assessments showed higher score in gene enhanced Mosaicplasty group. Glycosaminoglycan assay revealed no difference between groups involving Mosaicplasty. MRI analysis demonstrated the healing process between the subchondral bone other than control groups. CONCLUSIONS: hIGF-I gene enhanced tissue engineering can modify the outcome of Mosaicplasty to reconstruct large osteochondral defects in weight-bearing region.

A novel small heat shock protein gene, vis1, contributes to pectin depolymerization and juice viscosity in tomato fruit.

We have characterized a novel small heat shock protein gene, viscosity 1 (vis1) from tomato (Lycopersicon esculentum) and provide evidence that it plays a role in pectin depolymerization and juice viscosity in ripening fruits. Expression of vis1 is negatively associated with juice viscosity in diverse tomato genotypes. vis1 exhibits DNA polymorphism among tomato genotypes, and the alleles vis1-hta (high-transcript accumulator; accession no. AY128101) and vis1-lta (low transcript accumulator; accession no. AY128102) are associated with thinner and thicker juice, respectively. Segregation of tomato lines heterogeneous for vis1 alleles indicates that vis1 influences pectin depolymerization and juice viscosity in ripening fruits. vis1 is regulated by fruit ripening and high temperature and exhibits a typical heat shock protein chaperone function when expressed in bacterial cells. We propose that VIS1 contributes to physiochemical properties of juice, including pectin depolymerization, by reducing thermal denaturation of depolymerizing enzymes during daytime elevated temperatures.