ABSTRACT
Sweet potato is an important food crop that can also be used as an industrial raw material. Sucrose is the main form of long-distance carbohydrate transport in plants, and sucrose transporter (SUT) regulates the transmembrane transport and distribution of sucrose during plant growth and metabolism. Moreover, SUT plays a key role in phloem mediated source-to-sink sucrose transport and physiological activities, supplying sucrose for the sink tissues. In this study, the full-length cDNA sequences of IbSUT62788 and IbSUT81616 were obtained by rapid amplification of cDNA ends (RACE) cloning according to the transcripts of the two SUT coding genes which were differentially expressed in sweet potato storage roots with different starch properties. Phylogenetic analysis was performed to clarify the classification of IbSUT62788 and IbSUT81616. The subcellular localization of IbSUT62788 and IbSUT81616 was determined by transient expression in Nicotiana benthamiana. The function of IbSUT62788 and IbSUT81616 in sucrose and hexose absorption and transport was identified using yeast functional complementarity system. The expression pattern of IbSUT62788 and IbSUT81616 in sweet potato organs were analyzed by real-time fluorescence quantitative PCR (RT-qPCR). Arabidopsis plants heterologous expressing IbSUT62788 and IbSUT81616 genes were obtained using floral dip method. The differences in starch and sugar contents between transgenic and wild-type Arabidopsis were compared. The results showed IbSUT62788 and IbSUT81616 encoded SUT proteins with a length of 505 and 521 amino acids, respectively, and both proteins belonged to the SUT1 subfamily. IbSUT62788 and IbSUT81616 were located in the cell membrane and were able to transport sucrose, glucose and fructose in the yeast system. In addition, IbSUT62788 was also able to transport mannose. The expression of IbSUT62788 was higher in leaves, lateral branches and main stems, and the expression of IbSUT81616 was higher in lateral branches, stems and storage roots. After IbSUT62788 and IbSUT81616 were heterologously expressed in Arabidopsis, the plants grew normally, but the biomass increased. The heterologous expression of IbSUT62788 increased the soluble sugar content, leaf size and 1 000-seed weight of Arabidopsis plants. Heterologous expression of IbSUT81616 increased starch accumulation in leaves and root tips and 1 000-seed weight of seeds, but decreased soluble sugar content. The results obtained in this study showed that IbSUT62788 and IbSUT81616 might be important genes regulating sucrose and sugar content traits in sweet potato. They might carry out physiological functions on cell membrane, such as transmembrane transport of sucrose, sucrose into and out of sink tissue, as well as transport and unloading of sucrose into phloem. The changes in traits result from their heterologous expression in Arabidopsis indicates their potential in improving the yield of other plants or crops. The results obtained in this study provide important information for revealing the functions of IbSUT62788 and IbSUT81616 in starch and glucose metabolism and formation mechanism of important quality traits in sweet potato.
Subject(s)
Ipomoea batatas/metabolism , Arabidopsis/metabolism , Sucrose/metabolism , Saccharomyces cerevisiae/metabolism , DNA, Complementary , Phylogeny , Plants, Genetically Modified/genetics , Membrane Transport Proteins/metabolism , Starch/metabolism , Plant Proteins/metabolism , Gene Expression Regulation, PlantABSTRACT
Chemotherapy is among the limited choices approved for the treatment of hepatocellular carcinoma (HCC) at intermediate and advanced stages. Preferential and prolonged drug exposure in diseased sites is required to maximize the therapeutic index of the drug. Here, we report an injectable supramolecular peptide hydrogel as an intraperitoneal depot for localized and sustained release of triptolide for the treatment of orthotopic HCC. We chose peptide amphiphile C-GNNQQNYKD-OH-based nanofibers as gelators and carriers for triptolide. Sustained triptolide release from the hydrogel was achieved over 14 days , with higher accumulation in and cytotoxicity against human HCC Bel-7402 in comparison with L-02 fetal hepatocytes. After intraperitoneal injection, the hydrogel showed prolonged retention over 13 days and preferential accumulation in the liver, realizing HCC growth inhibition by 99.7 ± 0.1% and animal median survival extension from 19 to 43 days, without causing noticeable pathological changes in the major organs. These results demonstrate that injectable peptide hydrogel can be a potential carrier for localized chemotherapy of HCC.
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Objective@#To investigate the effects of emodin on lipopolysaccharide (LPS)-induced inflammation and damage in HK-2 cells.@*Methods@#The HK-2 cells were divided into blank group, emodin group, LPS group, and LPS+emodin group. The emodin group was treated with culture medium containing 0.5 μg/ml emodin, the LPS group was treated with culture medium containing 10 μg/ml LPS, the LPS+emodin group was treated with culture medium containing both 0.5 μg/ml emodin and 10 μg/ml LPS, and the blank group was cultured with fresh medium. Twelve hours later, HK-2 cells from each group were harvested for the analysis. The mRNA levels of oligonucleotide receptor protein 3 (NLRP3), IL-1β, IL-18, TNF-α, and neutrophil gelatinase-associated lipocalin (NGAL) in each group were determined by RT-PCR.@*Results@#Compared to the blank group, the mRNA levels of NLRP3 (2.74 ± 0.38 vs. 1.00 ± 0.14), IL-1β (2.40 ± 0.33 vs. 1.00 ± 0.19), and IL-18 (3.00 ± 0.42 vs. 1.00 ± 0.07), NGAL (31.73 ± 3.41 vs. 1.00 ± 0.07), and TNF-α (9.73±1.60 vs. 1.00 ± 0.11) significantly increased by LPS stimulation (P<0.05). Compared to the LPS group, the mRNA expression of NLRP3 (1.98 ± 0.24 vs. 2.74 ± 0.39), IL-1β (1.54 ± 0.24 vs. 2.40 ± 0.33), and IL-18 (2.09 ± 0.53 vs. 3.00 ± 0.42), NGAL (16.76 ± 1.72 vs. 31.73 ± 3.41) and TNF-α (9.73 ± 1.60 vs. 4.65 ± 1.09) in HK-2 cells of LPS+emodin group significantly decreased (P<0.05).@*Conclusions@#The emodin protects HK-2 cells from LPS-induced damage and inflammation by inhibiting NLRP3 inflammasome activation.
ABSTRACT
Objective To investigate the effects of emodin on lipopolysaccharide (LPS)-induced inflammation and damage in HK-2 cells. Methods The HK-2 cells were divided into blank group, emodin group, LPS group, and LPS+emodin group. The emodin group was treated with culture medium containing 0.5 μg/ml emodin, the LPS group was treated with culture medium containing 10 μg/ml LPS, the LPS+emodin group was treated with culture medium containing both 0.5 μg/ml emodin and 10 μg/ml LPS, and the blank group was cultured with fresh medium. Twelve hours later, HK-2 cells from each group were harvested for the analysis. The mRNA levels of oligonucleotide receptor protein 3 (NLRP3), IL-1β, IL-18, TNF-α, and neutrophil gelatinase-associated lipocalin (NGAL) in each group were determined by RT-PCR. Results Compared to the blank group, the mRNA levels of NLRP3 (2.74 ± 0.38 vs. 1.00 ± 0.14), IL-1β (2.40 ± 0.33 vs. 1.00 ± 0.19), and IL-18 (3.00 ± 0.42 vs. 1.00 ± 0.07), NGAL (31.73 ± 3.41 vs. 1.00 ± 0.07), and TNF-α (9.73±1.60 vs. 1.00 ± 0.11) significantly increased by LPS stimulation (P<0.05). Compared to the LPS group, the mRNA expression of NLRP3 (1.98 ± 0.24 vs. 2.74 ± 0.39), IL-1β (1.54 ± 0.24 vs. 2.40 ± 0.33), and IL-18 (2.09 ± 0.53 vs. 3.00 ± 0.42), NGAL (16.76 ± 1.72 vs. 31.73 ± 3.41) and TNF-α (9.73 ± 1.60 vs. 4.65 ± 1.09) in HK-2 cells of LPS+emodin group significantly decreased (P<0.05). Conclusions The emodin protects HK-2 cells from LPS-induced damage and inflammation by inhibiting NLRP3 inflammasome activation.
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3D printing technology has unique advantages and broad application prospect in biomedicine field. In recent years, cell printing, tissue printing, and organ printing technologies h have appeared successively, and drug printing and medical device printing have been realized. For complex surgical cases, surgeons and researchers have explored a surgical program that involves 3D printing technology, and completed many clinical applications including case discussions, surgical simulations and implantation surgeries. These efforts promoted the application and development of 3D printing technology in medical field. The purpose of this paper is to describe the application and research status of 3D printing technology in medical field from the aspects of medical teaching, orthopedic surgery, stomatology, bioprinting, drug printing, medical device manufacturing, etc., and put forward the prospects for future development.
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Objective To study the generating algorithm of personalized external fixation model based on STL file, so as to realize the fabrication of personalized external fixation through 3D scanning, model generation and 3D printing. Compared with the traditional plaster fixation, the external fixation obtained by the proposed method has the advantages of good adhesion to the external surface of the limb, good gas permeability and light weight. Methods In order to generate a personalized external fixation model, the key generating algorithms of the model were studied. The point cloud file of the residual limb was obtained by a 3D scanner, and then was converted into an STL format file. The triangular patches were processed by cutting, offsetting, triangulating and hollowing to create an external fixation model with good gas permeability and conformable surface that is fully fitted with the body surface. Finally, the external fixation was fabricated by 3D printing. Results Using the improved point offset algorithm, the problem of severe distortion of the point offset at the plane and sharp corners was solved. Using the tangent sort method, the points on the contour ring were sorted clockwise to obtain a triangular profile. The hollowing of the model was achieved by using the typical three-dimensional "cutting tool" mathematical model and the STL file space intersection method. Conclusions The improved model generating algorithm was validated based on a wrist case, and the personalized external fixed fixation model with conformable surface was obtained. It is proved that the proposed model generating algorithm is effective and practical.
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Objective:To observe the changes of metabolic intensity value of points on the face before and after needling bilateral Hegu (LI 4) in healthy people and provide scientific basis for association between Hegu (LI 4) and face/mouth. Methods:A total of 45 healthy college students were selected in this study. Using medical thermography and Pennes bio-heat transfer model, the infrared thermograph images on the face before and after needling bilateral Hegu (LI 4) were collected to observe the distribution of metabolic intensity value on the face before acupuncture and changes in these values after needling bilateral Hegu (LI 4). Results:Before acupuncture, Cuanzhu (BL 2) had the maximal metabolic intensity value. Its mean value was (0.71±0.23) W. Quanliao (SI 18) had the minimal metabolic intensity value. There were no left-right statistical significances in metabolic intensity values. After needling bilateral Hegu (LI 4), the metabolic intensity values of most points on the face were increased. Kouheliao (LI 19) obtained the maximal increase: 0.35 W on average; and Yangbai (GB 14) obtained the minimal increase: 0.08 W on average. Conclusion:Points on both sides in healthy people have good symmetry in metabolic intensity value. After needling bilateral Hegu (LI 4), the metabolic intensity values of points on the face were increased, especially points around the lips, which accords with the pathway of the Large Intestine Meridian on the head and face. This provided some scientific foundation for the association between Hegu (LI 4) and face/mouth.