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1.
Zhongguo Zhen Jiu ; 40(10): 1081-4, 2020 Oct 12.
Artigo em Chinês | MEDLINE | ID: mdl-33068350

RESUMO

OBJECTIVE: To evaluate the clinical effect of acupoint injection of houttuynia cordata as the accessory treatment on dry eyes of convalescent herpes simplex keratitis (HSK). METHODS: A total of 60 patients with dry eyes of convalescent HSK were randomized into an observation group and a control group, 30 cases in each one. In the control group, the artificial tears and anti-inflammatory drugs were combined in treatment. In the observation group, on the base of the treatment as the control group, the acupoint injection of houttuynia cordata at Neiqiuhou (Extra) was combined, 3 mL each time, once a day. After consecutive 3 injections, the injection was adjusted to be once every two days, consecutively for 3 times. The treatment for 6 times was as one course and one course of treatment was required. Separately, before treatment and in 7, 15 and 30 days after treatment, the changes of the scores of visual analogue scale (VAS), theresults of SchirmerⅠtest (SⅠT), the tear break-up time (BUT) and the score of corneal fluorescein staining (CFS) were observed and analyzed in the patients of the two groups. RESULTS: In 7, 15 and 30 days after treatment, VAS scores and CFS scores were all reduced as compared with those before treatment in the patients of the two groups (P<0.05), and the scores of VAS and CFS in the observation group were lower than those in the control group (P<0.05). In 7, 15 and 30 days after treatment, the values of SⅠT and BUT were all increased as compared with those before treatment in the patients of the two groups (P<0.05), and the values in the observation group were higher than the control group in 15 and 30 days after treatment (P<0.05). CONCLUSION: Acupoint injection of houttuynia cordata promotes corneal epithelial recovery, reduces the discomfort symptoms as well as increases tear secretion and the stability of tear film in dry eyes of convalescent herpes simplex keratitis.

2.
Carbohydr Polym ; 250: 116917, 2020 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-33049889

RESUMO

The present research was designed to fabricate a novel pH-responsive microgel based on chitosan/sodium alginate (CS/ALG) to encapsulate the polyphenols from the shells of Juglans regia L. (JRP-Microgel), and investigate the physicochemical properties of JRP-Microgel as well as analyze the release characteristic and phenolic composition variation under simulated gastrointestinal digestion in vitro. The results showed that JRP-Microgel belonged to semi-crystalline substances, and presented regular spherical or subspherical shape with neat edge and uniform particle size. JRP-Microgel possessed better physicochemical stabilities compared with unencapsulated JRP. Furthermore, JRP-Microgel exhibited excellent sustained release characteristic under simulated gastrointestinal digestion. Nevertheless, during different digestive phase, the phenolic composition of microgel was significantly different from the one of untreated JRP, suggesting that JRP-Microgel could prevent the degradation of JRP induced by gastrointestinal digestion. In conclusion, above results provide the experimental basis for the application of microgel encapsulation to improve the bioavailability of phenolic compounds in human body.

3.
Sleep Breath ; 2020 Oct 12.
Artigo em Inglês | MEDLINE | ID: mdl-33044690

RESUMO

PURPOSE: This study aimed to examine sleep quality and risk factors associated with sleep disturbance in patients undergoing total joint arthroplasty (TJA) managed with an enhanced recovery protocol from preoperatively until 12 weeks postoperatively in order to provide suggestions for improving the prevention and treatment of sleep disturbance in these patients. METHODS: The Chinese versions of the Pittsburgh Sleep Quality Index (CPSQI) and Epworth Sleepiness Scale (CESS) were used to evaluate sleep quality. Pain was measured by a visual analog scale (VAS). Neuropsychological status was also assessed, using the Center for Epidemiological Studies Depression Scale (CESD) and Zung Self-Rating Anxiety Scale (ZSAS). RESULTS: Of 107 patients (66% women), the prevalence of sleep disturbance was as high as 60% preoperatively. CPSQI score significantly increased postoperatively from baseline to 2 weeks before decreasing rapidly to a lower level than preoperative baseline. CESS scores of postoperative weeks 1 and 2 increased significantly compared with baseline and decreased significantly after 4 weeks, and a positive correlation was found between CESS and CPSQI score. VAS score significantly decreased from baseline over all time points in the postoperative period. A good sleeper group was more likely to suffer from severe sleep disturbance after surgery than a poor sleeper group with a bigger increase of CPSQI score. Multivariate logistic regression analysis indicated that pain and anxiety were risk factors associated with postoperative sleep disturbance. CONCLUSIONS: Sleep disturbance is highly prevalent among patients undergoing TJA. More attention and multimodal approaches with well-controlled pain, mental counseling, and possibly preemptive use of sleep medication may improve sleep quality in TJA patients to promote recovery.

4.
Plants (Basel) ; 9(10)2020 Oct 06.
Artigo em Inglês | MEDLINE | ID: mdl-33036187

RESUMO

Water shortages have an important impact on the photosynthetic capacity of Phragmites australis. However, this impact has not been adequately studied from the perspective of photosynthesis. An in-depth study of the photosynthetic process can help in better understanding the impact of water shortages on the photosynthetic capacity of P. australis, especially on the microscale. The aim of this study is to explore the photosynthetic adaptation strategies to environmental changes in saline‒alkaline wetlands. The light response curves and CO2 response curves of P. australis in five habitats (hygrophilous, xerophytic, psammophytic, abandoned farmland, paddy field drainage) in saline‒alkaline wetlands were measured at different stages of their life history, and we used a nonrectangular hyperbolic model to fit the data. It was concluded that P. australis utilized coping strategies that differed between the growing and breeding seasons. P. australis in abandoned farmland during the growing season had the highest apparent quantum efficiency (AQE) and photosynthetic utilization efficiency for weak light because of the dark environment. The dark respiration rate of P. australis in the drainage area of paddy fields was the lowest, and it had the highest values for photorespiration rate, maximum photosynthetic rate (Pmax), photosynthetic capacity (Pa), biomass, maximum carboxylation rate (Vcmax), and maximum electron transfer rate (Jmax). The light insensitivity of P. australis increased with the transition from growing to breeding season, and the dark respiration rate also showed a downward trend. Moreover, Vcmax and Jmax would decline when Pmax and Pa showed a declining trend, and vice versa. In other words, Vcmax and Jmax could explain changes in the photosynthetic capacity to some extent. These findings contribute to providing insights that Vcmax and Jmax can directly reflect the variation in photosynthetic capacity of P. australis under water shortages in saline‒alkaline wetlands and in other parts of world where there are problems with similarly harmful environmental conditions.

5.
Nat Commun ; 11(1): 4947, 2020 10 02.
Artigo em Inglês | MEDLINE | ID: mdl-33009392

RESUMO

Pseudomonas syringae is a Gram-negative and model pathogenic bacterium that causes plant diseases worldwide. Here, we set out to identify binding motifs for all 301 annotated transcription factors (TFs) of P. syringae using HT-SELEX. We successfully identify binding motifs for 100 TFs. We map functional interactions between the TFs and their targets in virulence-associated pathways, and validate many of these interactions and functions using additional methods such as ChIP-seq, electrophoretic mobility shift assay (EMSA), RT-qPCR, and reporter assays. Our work identifies 25 virulence-associated master regulators, 14 of which had not been characterized as TFs before.

6.
Cancer Lett ; 495: 200-210, 2020 Sep 12.
Artigo em Inglês | MEDLINE | ID: mdl-32931885

RESUMO

In addition to their hypoglycemic effect, sodium-glucose cotransporter 2 (SGLT2) inhibitors have many other benefits. In the present study, we examine the anticancer effect of the SGLT2 inhibitor empagliflozin using cervical carcinoma models. In vivo antitumor activities of empagliflozin were observed in a nude mouse model. Empagliflozin intervention and downregulation of Sonic Hedgehog Signaling Molecule (Shh) inhibited the migration and promoted the apoptosis of cervical cancer cells in nude mice. Compared with the control group, the empagliflozin treatment group had an increased level of AMP-activated protein kinase (AMPK) and decreased levels of Forkhead Box A1 (FOXA1) and SHH in tumor tissue. In vitro experiments also showed that empagliflozin (50 µM) inhibited the migration of cervical cancer cells and induced their apoptosis by activating the AMPK/FOXA1 pathway and inhibiting the expression of SHH. Kaplan-Meier survival analysis was used to determine the relationship between SHH expression and total survival time. The results showed that in cervical cancer patients, high SHH expression resulted in unfavorable overall survival. The downregulation of SHH with small interfering RNA (siRNA) inhibited the migration and invasion and promoted the apoptosis of HeLa cells. These findings show that empagliflozin has a potential therapeutic effect on cervical cancer. This effect was related to the activation of the AMPK pathway and the inhibition of SHH expression.

7.
Mol Med Rep ; 22(5): 3635-3644, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32901841

RESUMO

Long non­coding RNAs (lncRNAs) and microRNAs (miRs) have been reported to regulate disease progression in numerous types of disease, including retinoblastoma (Rb). Therefore, the present study aimed to investigate the effects of the lncRNA FEZ family zinc finger 1 antisense RNA 1 (FEZF1­AS1) on Rb and to determine its possible mechanism of action. Reverse transcription­quantitative PCR and western blot analysis were conducted to detect the gene or protein expression. Cell Counting Kit­8, wound healing and transwell invasion assays were performed to estimate the capabilities of cell viability, invasion and migration. The potential association between FEZF1­AS1 and miR­1236­3p in Y79 cells was measured via dual­luciferase reporter assay. The results of the present study revealed that the levels of FEZF1­AS1 were significantly upregulated in different Rb cell lines, with the most prominent upregulation observed in Y79 cells. In addition, the cell viability, invasive and migratory abilities, and the ability to undergo epithelial­mesenchymal transition (EMT), were significantly inhibited following the transfection of short hairpin RNA (shRNA)­FEZF1­AS1 into Y79 cells. Further experimental validation confirmed that miR­1236­3p may be a direct target of FEZF1­AS1. Notably, the miR­1236­3p inhibitor was discovered to reverse the inhibitory effects of shRNA­FEZF1­AS1 on cell viability, invasion, migration and EMT. In conclusion, the findings of the present study suggested that lncRNA­FEZF1­AS1 may promote the viability, migration, invasion and EMT of Rb cells by modulating miR­1236­3p.

8.
J Bone Miner Metab ; 2020 Sep 02.
Artigo em Inglês | MEDLINE | ID: mdl-32876727

RESUMO

INTRODUCTION: Bone turnover markers (BTMs) can be used to monitor bone metabolism, while the actual clinical changing in hip fracture had not been certified to evaluate the changes of BTMs during the healing process after surgery of elderly hip fractures; and to get the effects of operation type, gender, serum 25(OH)D level, and age on bone turnover markers. MATERIALS AND METHODS: A total of 100 elderly cases with hip fracture were selected, including 74 females and 26 males, and the patients were followed to 180-230 days after surgery. Serum levels of N-propeptide of type 1 collagen (P1NP), C-terminal crosslinking telopeptides of type 1 collagen (CTX), Osteocalcin (OC), and 25 hydroxy vitamin D (25OHD) were investigated. Bone mineral density (BMD) was measured with dual-energy X-ray absorptiometry (DXA). RESULTS: (1) P1NP and CTX showed peak time at 30-60 days after operation, while OC keep going even at 180-230 days; P1NP showed less than 4 times elevation during healing, CTX and OC only had less than 2 times rise. (2) Female had higher serum CTX and OC than male, intramedullary nailing for intertrochanteric fracture patients had higher P1NP than hip replacement for femoral neck fracture patients, and both the degrees of increase were less than 50%. (3) Serum average 25(OH)D level had no effect on BTMs during the fracture healing; different from the young old (65-84 years), serum OC level of eldest older patients(≥ 85 years) decreased early in the process of fracture healing. CONCLUSIONS: BTMs reached the peak level in 30-60 days after surgery, P1NP showed less than 4 times elevation, and CTX and OC had less than 2 times rise. It was not necessary to take gender into account when observing P1NP, and it was not necessary to take fracture and operation type into account when observing CTX and OC.

9.
Small ; : e2003834, 2020 Sep 02.
Artigo em Inglês | MEDLINE | ID: mdl-32877017

RESUMO

Cell adhesion of nanosystems is significant for efficient cellular uptake and drug delivery in cancer therapy. Herein, a near-infrared (NIR) light-driven biomimetic nanomotor is reported to achieve the improved cell adhesion and cellular uptake for synergistic photothermal and chemotherapy of breast cancer. The nanomotor is composed of carbon@silica (C@SiO2 ) with semi-yolk@spiky-shell structure, loaded with the anticancer drug doxorubicin (DOX) and camouflaged with MCF-7 breast cancer cell membrane (i.e., mC@SiO2 @DOX). Such biomimetic mC@SiO2 @DOX nanomotors display efficient self-thermophoretic propulsion due to a thermal gradient generated by asymmetrically spatial distribution. Moreover, the MCF-7 cancer cell membrane coating can remarkably reduce the bioadhesion of nanomotors in biological medium and exhibit highly specific self-recognition of the source cell line. The combination of effective propulsion and homologous targeting dramatically improves cell adhesion and the resultant cellular uptake efficiency in vitro from 26.2% to 67.5%. Therefore, the biomimetic mC@SiO2 @DOX displays excellent synergistic photothermal and chemotherapy with over 91% MCF-7 cell growth inhibition rate. Such smart design of the fuel-free, NIR light-powered biomimetic nanomotor may pave the way for the application of self-propelled nanomotors in biomedicine.

10.
BMC Plant Biol ; 20(1): 406, 2020 Sep 02.
Artigo em Inglês | MEDLINE | ID: mdl-32878602

RESUMO

BACKGROUND: Oat (Avena sativa L.) is a recognized health-food, and the contributions of its different candidate A-genome progenitor species remain inconclusive. Here, we report chloroplast genome sequences of eleven Avena species, to examine the plastome evolutionary dynamics and analyze phylogenetic relationships between oat and its congeneric wild related species. RESULTS: The chloroplast genomes of eleven Avena species (size range of 135,889-135,998 bp) share quadripartite structure, comprising of a large single copy (LSC; 80,014-80,132 bp), a small single copy (SSC; 12,575-12,679 bp) and a pair of inverted repeats (IRs; 21,603-21,614 bp). The plastomes contain 131 genes including 84 protein-coding genes, eight ribosomal RNAs and 39 transfer RNAs. The nucleotide sequence diversities (Pi values) range from 0.0036 (rps19) to 0.0093 (rpl32) for ten most polymorphic genes and from 0.0084 (psbH-petB) to 0.0240 (petG-trnW-CCA) for ten most polymorphic intergenic regions. Gene selective pressure analysis shows that all protein-coding genes have been under purifying selection. The adjacent position relationships between tandem repeats, insertions/deletions and single nucleotide polymorphisms support the evolutionary importance of tandem repeats in causing plastome mutations in Avena. Phylogenomic analyses, based on the complete plastome sequences and the LSC intermolecular recombination sequences, support the monophyly of Avena with two clades in the genus. CONCLUSIONS: Diversification of Avena plastomes is explained by the presence of highly diverse genes and intergenic regions, LSC intermolecular recombination, and the co-occurrence of tandem repeat and indels or single nucleotide polymorphisms. The study demonstrates that the A-genome diploid-polyploid lineage maintains two subclades derived from different maternal ancestors, with A. longiglumis as the first diverging species in clade I. These genome resources will be helpful in elucidating the chloroplast genome structure, understanding the evolutionary dynamics at genus Avena and family Poaceae levels, and are potentially useful to exploit plastome variation in making hybrids for plant breeding.

11.
Sheng Wu Gong Cheng Xue Bao ; 36(8): 1546-1555, 2020 Aug 25.
Artigo em Chinês | MEDLINE | ID: mdl-32924353

RESUMO

Sucrose phosphorylase (SPase) gene from Leuconostoc mesenteroides ATCC 12291 was synthesised after codon optimization, and inserted into pET-28a plasmid to generate pET-28a-spase. The recombinant strain Escherichia coli BL21 (DE3)/pET-28a-spase was induced for Spase expression. The recombinant protein Spase was purified and characterized. The specific enzyme activity of SPase was 213.98 U/mg, the purification ratio was 1.47-fold, and the enzyme activity recovery rate was 87.80%. The optimal temperature and the optimal pH of the SPase were identified to be 45 °C and 6.5 respectively, and Km, Vmax and kcat of the SPase for sucrose was 128.8 mmol/L, 2.167 µmol/(mL·min), and 39 237.86 min-1. The recombinant SPase was used for α-arbutin production from hydroquinone and the reaction process was evaluated. The optimal conditions for synthesis of α-arbutin by SPase were 40 g/L hydroquinone, 5:1 molar ratio of sucrose and hydroquinone, and 250 U/mL recombinant SPase at pH 7.0 and 30 °C for 24 h in the dark, and then 500 U/mL glucoamylase was added at 40°C for 2.5 h. Under the optimized process, the yield of α-arbutin reached 98 g/L, and the hydroquinone conversion rate was close to 99%. In summary, the recombinant SPase was cloned and characterized, and its application for α-arbutin production was feasible.

12.
Artigo em Inglês | MEDLINE | ID: mdl-32962860

RESUMO

Phosphate transporters (PHTs) are well-known for their roles in phosphate uptake in plants. However, their actions in imparting plant growth in plants are still not so clear. In our previous study, we observed that maize PHT1 gene ZmPt9 plays a significant role in phosphate uptake. In this study, we further characterized ZmPt9 in response to low phosphate condition through ZmPt9 promoter inductive analysis by GUS staining and quantification. To elucidate the function of ZmPt9, we generated overexpression plant in Arabidopsis. ZmPt9 overexpressing Arabidopsis plants conferred small leaves and early flowering compared with the wild-type plants. In addition, ZmPt9 can complement the late flowering phenotype of Arabidopsis mutant pht1;2. The qRT-PCR analysis revealed that overexpression of ZmPt9 in Arabidopsis changed expression levels of some flowering-related genes. Further expressed detection of hormone related genes revealed that GA and auxin maybe the main determinant for growth influences of ZmPt9. In conclusion, these results suggest that apart from phosphate transport activity, ZmPt9 can be further exploited for improving crops growth.

13.
Int J Biol Macromol ; 2020 Sep 14.
Artigo em Inglês | MEDLINE | ID: mdl-32941904

RESUMO

Understanding the structure and properties of lignin has important practical significance for its further applications. In this case, eucalyptus was fractionated with 88% formic acid at 101 °C for different durations, and the removal efficiency as well as the chemical structure of lignin at various stages were comparatively analyzed. The obtained data indicated that with increasing reaction time, lignin was continuously removed and the process could be divided into three stages. The lignin dissolution rate was fast first and then slow, and the molecular weight of the dissolved lignin increased with time. The lignin structure was condensed and the molecular weight increased with prolonged of reaction time. Structural analysis indicated that the ß-O-4' structure was largely destroyed, the G-type lignin dissolved early, and the degradation of the S-type lignin became more intensive with increasing reaction time. This is of great help for reaction control as well as the further processing of lignin byproducts.

14.
J Chromatogr Sci ; 2020 Aug 31.
Artigo em Inglês | MEDLINE | ID: mdl-32869056

RESUMO

A new capillary electrophoresis method was applied to chiral separation of three amino acids, including D,L-tryptophan, D,L-tyrosine and D,L-phenylalanine. The chiral resolution was attained in an untreated fused-sillica capillary using a dual chiral selector, which was made up of Cu(II)-L-histidine complex and ß-cyclodextrin (CD). The cardinal factors influencing its separation efficiency, such as chiral selectors, buffer pH and applied voltage, were optimized. Best results were acquired by using a buffer consisting of 10 mmol/L Cu(II), 13 mmol/L L-histidine, 8 mmol/L ß-CD, 5 mmol/L phosphate adjusted to pH 5.0 and 15 kV applied voltage. All enantiomers were entirely resolved within 20 min with high resolutions of 3.6~6.1. The analysis method was verified through the determination of D,L-tryptophan in terms of linearity, precision and accuracy. And the robustness of this method was proved. The Limit of Detection and Limit of Quantification for both enantiomers were 2.5 and 5 µg/mL, respectively. The method was perfectly applied to the determination of the enantiomeric purity of L-tryptophan. Furthermore, the interaction between Cu(II)-L-histidine complex and ß-CD was also studied using Ultraviolet-visible and 1H NMR spectroscopy to explain the synergistic effect involved. The results illustrated that Cu(II)-L-histidine complex and ß-CD played a synergistic role in the enantiomeric separation of chiral drugs, with good prospects for application.

15.
J Clin Lab Anal ; : e23549, 2020 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-32869917

RESUMO

BACKGROUND: As one of the most prevalent diagnostic indicators of diabetic kidney disease (DKD), albumin-to-creatinine ratio (ACR) shows considerably limited predictive power in clinical application. We analyzed microarray expression profiling of urine to seek for differentially expressed miRNAs for potential biomarkers of DKD. METHODS: Urine samples from type 2 diabetes mellitus (T2DM) patients with (30 mg/g < ACR < 300 mg/g, DKD group) or without DKD (ACR < 30 mg/g, DM group) were collected for miRNA microarray analysis. The differentially expressed miRNAs were screened by bioinformatics analysis and validated by quantitative real-time PCR. Target genes of differentially expressed miRNAs were predicted in miRDB, Targetscan, and microRNA.org databases. We also conducted the Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathways analysis to explore for potential mechanisms in DKD. RESULTS: Nine miRNAs were down-regulated and seventeen miRNAs were up-regulated in DKD group, compared to DM group. The levels of miR-3137 and miR-4270 in DKD group were 0.670 ± 0.505 and 2.116 ± 1.762 times than those in DM group, respectively, showing great significance. A total of 1076 target genes were simultaneously predicted by miRDB, Targetscan, and microRNA.org databases. According to the GO analysis results, disorders of endomembrane system may be one of the major pathological changes in DKD. In addition, Rap 1 signaling pathway is also altered obviously in DKD, discovered by the KEGG analysis. CONCLUSION: MiR-3137 and miR-4270 show the potential for urinary biomarkers of DKD. The pathological changes of DKD may be related to disorders of endomembrane system and alternation of Rap1 signaling pathway.

16.
Talanta ; 219: 121346, 2020 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-32887075

RESUMO

N-glycan modification is reported to be important in regulating the structure and function of immunoglobulins in mammals. While, the study on teleost immunoglobulin glycosylation is still limitted. In this study, we constructed a TNP-antigen driven model, and detected the site-specific N-glycans of PBS-immunized and TNP-specific Oreochromis niloticus serum IgM through 18O-labeling and nanoLC-MS/MS. These methods are widely used for peptide enrichment and protein modification identification, but rarely used in detecting the level of N-glycosylation in teleost Igs that driven by specific antigen. The results revealed that there are four N-glycosylation sites in O.niloticus IgM heavy chain, namely, the Asn-315 site in the CH2 domain, the Asn-338 site in the CH3 domain, and the Asn-509 and Asn-551 sites in the CH4 domain, All of the four residues were efficiently N-glycosylated. After immunized with TNP-antigen, the signal strength of oligomannose in the TNP-specific IgM in primary mass spectrometry was significantly higher than that in the PBS-immunized IgM. Notably, the TNP-specific IgM had an Asn-509 site fully occupied with oligomannose, while only a small amount of oligomannose was found in the PBS-immunized IgM of this site. N-glycans in other sites were mainly complex-type with a low content of fucosylation and sialylated. The oligomannose in TNP-specific IgM was further verified to be essential for the binding of IgM and MBL. These results demonstrated that the TNP-antigen induced the site-specific oligomannose modification of O.niloticus IgM heavy chain, and played an important role in the interaction of IgM and MBL, which provided insights into the evolutionary understanding of the IgM oligomannose modification and function.

17.
Nat Protoc ; 2020 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-32978602

RESUMO

Pseudotyped viruses are useful virological tools because of their safety and versatility. On the basis of a vesicular stomatitis virus (VSV) pseudotyped virus production system, we developed a pseudotyped virus-based neutralization assay against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in biosafety level 2 facilities. Compared with the binding antibody test, the neutralization assay could discriminate the protective agents from the antibody family. This protocol includes production and titration of the SARS-CoV-2 S pseudotyped virus and the neutralization assay based on it. Various types of samples targeting virus attachment and entry could be evaluated for their potency, including serum samples derived from animals and humans, monoclonal antibodies and fusion inhibitors (peptides or small molecules). If the pseudotyped virus stock has been prepared in advance, it will take 2 days to get the potency data for the candidate samples. Experience in handling cells is needed before implementing this protocol.

18.
Fish Shellfish Immunol ; 106: 609-620, 2020 Aug 12.
Artigo em Inglês | MEDLINE | ID: mdl-32795594

RESUMO

Edwardsiellosis, an extremely harmful disease can be caused by Edwardsiella tarda, severely restricts the development of turbot (Scophthalmus maximus) farming worldwide, especially in China. This study aimed to establish an effective and feasible prophylaxis by feeding chitosan-alginate coated egg yolk immunoglobulin (IgY) against E. tarda 2CDM001 infections in the process of turbot farming. Enzyme-linked immunosorbent assays proved that the obtained specific IgY could specifically target E. tarda 2CDM001 and five other E. tarda isolates (1a5p, Hz-s, 1a1s, fs-a1 and 58p8). In-vitro, the bacteriostatic effects of specific IgY showed dose dependencies at concentrations ranging from 1 to 10 mg/mL. Moreover, E. tarda 2CDM001 incubated with 10 mg/mL specific IgY could induce the destruction of cell wall structures and significantly decrease the bacterial surface hydrophobicity (p < 0.05). In this study, turbots were challenged with 107 CFU E. tarda 2CDM001 after seven days of continuous feeding with basal diets containing microencapsulated IgYs. Survival rates of the 5%, 3% and 1% microencapsulated specific IgY groups were 63.3%, 56.7% and 20% on the tenth day post infection, respectively, while the turbots in the positive control and non-specific IgY groups all died within ten days. Oral administration of basal diets containing 5% microencapsulated specific IgY significantly reduced IL-1ß, IL-8, TNF-α and C3 transcript levels in the head kidney and spleen of turbots compared with the positive and non-specific IgY groups at 24 h after E. tarda 2CDM001 challenging (p < 0.05). Pathological increase of leukocytes in the specific IgY group was significantly lower than that in the positive control and non-specific IgY groups (p < 0.05), decreasing slowly after 24 h of infection and showing a recovery trend. Erythrocyte counts and hemoglobin concentrations of turbots in positive and non-specific IgY groups showed a marked decrease compared with the negative and specific groups at 96 h after E. tarda 2CDM001 infection (p < 0.05). These results suggest that passive immunity via feeding microencapsulated specific IgY could be used as a valuable preventative in turbot against E. tarda 2CDM001 infections.

19.
Antonie Van Leeuwenhoek ; 113(11): 1573-1585, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32815093

RESUMO

Colletotrichum gloeosporioides is a main cause of rubber anthracnose, which results in a huge loss for the natural rubber industry. In this study, an actinomycete strain QY-3 was isolated and had good antagonistic activity against C. gloeosporioides with an inhibition rate of 86.6%. Strain QY-3 was identified as Streptomyces deccanensis preliminarily. Millet medium was selected as the optimal fermentation broth for antifungal metabolites production by S. deccanensis QY-3. The culture filtrate extract (CFE) from the millet broth of S. deccanensis QY-3 exhibits broad-spectrum antifungal activity against plant pathogenic fungi, and its EC50 inhibiting the mycelial growth of C. gloeosporioides is 6.3 µg/mL. The CFE has good thermal and pH stabilities, and it can break the hyphae and inhibit the conidial germination of C. gloeosporioides. 100 µg/mL of CFE had an obvious control effect on rubber anthracnose, and the control efficacy was 63.7% on 5 days after inoculation. Two compounds with inhibitory effects on C. gloeosporioides, anthranilic acid and sangivamycin, were isolated from the CFE. The MICs of both compounds against C. gloeosporioides were 29.3 and 23.0 µg/mL respectively. In conclusion, the CFE from S. deccanensis QY-3 has great potential to be a promising fungicide for rubber anthracnose.

20.
Metabolism ; 111: 154334, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32777444

RESUMO

BACKGROUND AND PURPOSE: Excessive mitochondrial fission was observed in diabetic kidney disease (DKD). Phosphoglycerate mutase family member 5 (PGAM5) plays an important role in mitochondrial fission by dephosphorylating the dynamin-related protein 1 at Ser637 (DRP1S637). Whether PGAM5 participates in the mitochondrial fission in diabetic renal tubular injury is unknown. Clinical trials have observed encouraging effect of Sodium-glucose cotransporter 2 (SGLT2) inhibitors on DKD though the underling mechanisms remain unclear. EXPERIMENTAL APPROACH: We used KK-Ay mice as diabetic model and Empagliflozin (Empa) were administrated by oral gavage. The mitochondrial fission and the expressions of phosphorylated AMP-activated protein kinase (p-AMPK), specificityprotein1 (SP1), PGAM5 and DRP1S637 were tested. We also examined these changes in HK2 cells that cultured in normal glucose (NG), high glucose (HG) and high glucose+Empa (HG + Empa) environment. Then we verified our deduction using AMPK activator (5-aminoimidazole-4-carboximide Riboside, AICAR), inhibitor (Compound C), si-SP1 and si-PGAM5. Lastly, we testified the interaction between SP1 and the PGAM5promotor by CHIP assay. KEY RESULTS: The mitochondrial fission and the expression of SP1, PGAM5 increased and the expression of p-AMPK, DRP1S637 decreased in diabetic or HG environment. These changes were all reversed in Empa or AICAR treated groups. These reversal effects of Empa could be diminished by Compound C. Either si-SP1 or si-PGAM5 could alleviate the mitochondrial fission without affection on AMPK phosphorylation. Finally, the CHIP assay confirmed the interaction between SP1 and the PGAM5 promotor. CONCLUSIONS AND IMPLICATIONS: The PGAM5 aggravated the development of diabetic renal tubular injury and the Empa could improve the DKD by alleviating mitochondrial fission via AMPK/SP1/PGAM5 pathway.

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