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1.
Plant Dis ; : PDIS05191033RE, 2020 Feb 13.
Artigo em Inglês | MEDLINE | ID: mdl-32053476

RESUMO

Pectobacterium carotovorum, a causal agent of vegetable soft rot, contains three valid subspecies: P. carotovorum subsp. carotovorum (Pcc), P. carotovorum subsp. brasiliensis (Pcb), and P. carotovorum subsp. odoriferum (Pco). Using 16S rDNA sequencing and genus-specific PCR, we identified 72 P. carotovorum strains from Chinese cabbage, bok choy, and celery and assessed their pathogenicity on Chinese cabbage petioles and potato tubers. Based on phylogenetic analysis of pmrA sequences and confirmation by subspecies-specific PCR, the strains were divided into 18 Pcc, 29 Pco, and 25 Pcb. Several characteristic features were also assessed and supported the distinctiveness of the Pco strains. All P. carotovorum strains caused soft rot symptoms on Chinese cabbage and potato, but the Pco strains exhibited the greatest severity. We developed a conventional PCR and a quantitative PCR (qPCR) assay for the identification of Pco based on its specific srlE gene encoding sorbitol-specific phosphotransferase. These two methods could specifically amplify the expected products of 674 and 108 bp, respectively, from all of the Pco strains. The assays demonstrated high sensitivity and could detect as little as 1 and 100 pg/µl of bacterial genomic DNA, respectively. Both assays could also detect the pathogens directly from plant tissues infected with as little as 2.5 × 10-2 CFU/mg of Pco, even before external symptoms appeared. These assays constitute effective tools for disease diagnosis and the rapid identification of soft rot pathogens.

2.
Int J Biol Macromol ; 149: 951-961, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-32018010

RESUMO

Phauda flammans Walker belongs to Phaudidae (Lepidoptera), which is a holometabolous and leaf-eating pest that harms trees. So far, there is no mitochondrial (mt) genome reported of Phaudidae. Herein, we sequenced and annotated the complete mt genome of P. flammans representing the first mt genome of Phaudidae and predicted the secondary structures of its RNAs in this study. This mt genome is 15470 bp long consisting of 13 protein coding genes (PCGs), 22 tRNAs, 2 rRNAs and the control region, which are usually conserved in insects. Most PCGs used the standard ATN start codons and complete TAA/TAG termination codons. Almost all of tRNA genes exhibited cloverleaf secondary structures except that the dihydorouridine (DHU) arm of tRNASer(AGN) was absent. The phylogenetic analyses using both Bayesian inference (BI) and maximum likelihood (ML) methods all supported that Phaudidae was a single family being the sister group to Zygaenidae. More mt genomes are needed to better understand the phylogenetic relationships within Zygaenoidea in the future.

3.
J Immunol ; 2020 Feb 07.
Artigo em Inglês | MEDLINE | ID: mdl-32034060

RESUMO

MHC class II (MHC-II) molecules play a crucial role in cellular and humoral immunity by forming peptide-MHC-II (pMHC-II) complexes. The three-dimensional structures of pMHC-II complexes have been well resolved in humans and mice. However, there is no structural information for pMHC-II complexes in nonmammals. In chickens, there are two closely related and highly polymorphic ß-chains and one monomorphic α-chain, and the mechanism by which one monomorphic α-chain combines with two polymorphic ß-chains to form a functional heterodimer remains unknown. In this study, we report the crystal structure of a chicken pMHC-II complex (pBL2*019:01) at 1.9-Å resolution as the first nonmammalian structure of a pMHC-II complex. The structure reveals an increase in hydrogen bonding between the α and ß main chains at the central interface that is introduced by the insertion of four residues in the α-chain. The residues in the ß-chain that form hydrogen bonds with the α-chain are conserved among all ß alleles. These structural characteristics explain the phenomenon of only one BLA allele without sequence variation pairing with highly diverse BLB alleles from two loci in the genome. Additionally, the characteristics of the peptide in the peptide-binding groove were confirmed. These results provide a new understanding of the pairing mechanism of the α- and ß-chains in a pMHC-II complex and establish a structural principle to design epitope-related vaccines for the prevention of chicken diseases.

4.
Int J Biol Macromol ; 2020 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-32035148

RESUMO

OBJECTIVE: Recombinant collagen (rCOL)-hyaluronic acid (HA) composite scaffolds were prepared to thoroughly investigate their biological properties. METHODS: The rCOL and HA composite scaffolds were formulated via lyophilization. The scaffolds were characterized for various materials properties, including porosity, surface modification, and degradation rates. Biological properties such as in vitro cytotoxicity, cell adhesion, proliferation and migration effects were also evaluated. RESULTS: The water absorption, mechanical strength, degradation resistance and thermal stability of the prepared rCOL-HA composites were improved over that of the control studied. Scanning electron microscopy (SEM) revealed that the composites formed a three-dimensional network structure with uniform pore distribution. The cytotoxicity of the composites was minimal (grade I) and the material showed strong adhesion and proliferation effects when grown with mouse fibroblasts, particularly the composite material of rCOL (5% HA) group (P < 0.05). CONCLUSION: The rCOL-HA composite prepared via lyophilization after cross-linking is characterized by high porosity, high water absorption, and good interaction between the material and cells, as well as good biodegradability. Compared with rCOL materials, rCOL-HA has increased mechanical strength, water absorption and thermal stability. The biocompatibility and fibroblast proliferation of rCOL-HA have excellent biological performance, providing a new material for wound healing applications.

5.
J Clin Lab Anal ; : e23224, 2020 Jan 31.
Artigo em Inglês | MEDLINE | ID: mdl-32004399

RESUMO

Thromboelastography (TEG) as a global coagulation test has been continuously developed for many decades in either research or clinical practice. The versatility of TEG test leads to difficulty in standardization and result interpretation. Reference intervals (RIs) of TEG may be one of the most controversial factors that influence its wide applications. RIs establishment with the traditional method is time-consuming and laborious as well as beyond general laboratory capability. Indirect method using stored data and with statistical calculation and small cost is emerging as an alternative approach for RIs determination. Gender, age, or both affect RIs and must be taken into account before RIs estimation. The present study retrospectively collected a total of 930 TEG results as subjects and established RIs with indirect method for Kaolin-activated TEG, including the parameters of R, K, αAngle, MA, and CI. Furthermore, gender, age, and gender-dependent age subsets analyses were performed to determine their effects on RIs of TEG. In this study, we found that TEG parameters showed more hypercoagulability in female than male, most of the measured TEG variables were significantly associated with aging, but only in male statistical significance was found among different age stratification and 60-year-old could be considered as cutting point to differentiate coagulation ability in male. In addition, RIs of TEG were estimated by indirect method suitably and verified to be valid in our study. Finally, the RIs of TEG by indirect method were basically significantly different to the RIs recommended by manufacturer, but the consistent percentage is relatively high in the most of measured parameters. In conclusion, it is suggestive that the indirect method for RIs establishment is feasible, but relevant factors, such as gender and age, specifically gender-dependent age effect, should be considered before RIs determinations.

6.
Aging (Albany NY) ; 122020 Feb 11.
Artigo em Inglês | MEDLINE | ID: mdl-32045883

RESUMO

Atherosclerosis is one of the most common clinical cardiovascular disorders. Accumulating evidence indicates that lncRNAs exert critical functions in atherosclerosis; however, their functional roles and regulatory mechanisms remain unclear. In this study, we induced atherosclerotic plaques in three rabbit carotid arteries through an atherogenic diet and balloon injury; three age-matched rabbits were fed normal chow and served as controls. We thoroughly investigated the RNA (mRNA, lncRNA and miRNA) expression profiles in atherosclerotic rabbit carotid models with deep RNA sequencing. We identified several significantly differentially expressed RNAs. The corresponding lncRNA-miRNA-mRNA network was constructed, and the significantly dysregulated network was selected. Furthermore, Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses indicated that the mRNAs in the network were involved in leukocyte activation, cell proliferation, cell adhesion molecules and cytokine-cytokine receptor interaction. After rigorous screening, we obtained a differentially expressed lncRNA-miRNA-mRNA interaction network associated with atherosclerosis. In the network, XLOC_054118 and XLOC_030217 upregulate the CHI3L1, SOAT, CTSB and CAPG genes by competitively binding to the miRNA ocu-miR-96-5p. XLOC_062719 and XLOC_063297 upregulate CTSS, CTSB and EDNRA genes by competitively binding to the miRNA ocu-miR-185-5p.

7.
Trials ; 21(1): 170, 2020 Feb 11.
Artigo em Inglês | MEDLINE | ID: mdl-32046760

RESUMO

BACKGROUND: Necrotizing enterocolitis (NEC) is the leading cause of death among preterm infants born at < 30 weeks' gestation. The incidence of NEC is reduced when infants are fed human milk. However, in many neonatal intensive care units (NICUs), it is standard practice to freeze and/or pasteurize human milk, which deactivates bioactive components that may offer additional protective benefits. Indeed, our pilot study showed that one feed of fresh mother's own milk per day was safe, feasible, and can reduce morbidity in preterm infants. To further evaluate the benefits of fresh human milk in the NICU, a randomized controlled trial is needed. METHODS: Our prospective multicenter, double-blinded, randomized, controlled trial will include infants born at < 30 weeks' gestation and admitted to one of 29 tertiary NICUs in China. Infants in the intervention (fresh human milk) group (n = 1549) will receive at least two feeds of fresh human milk (i.e., within 4 h of expression) per day from the time of enrollment until 32 weeks' corrected age or discharge to home. Infants in the control group (n = 1549) will receive previously frozen human milk following the current standard protocols. Following informed consent, enrolled infants will be randomly allocated to the control or fresh human milk groups. The primary outcome is the composite outcome mortality or NEC ≥ stage 2 at 32 weeks' corrected age, and the secondary outcomes are mortality, NEC ≥ stage 2, NEC needing surgery, late-onset sepsis, retinopathy of prematurity (ROP), bronchopulmonary dysplasia (BPD), weight gain, change in weight, increase in length, increase in head circumference, time to full enteral feeds, and finally, the number and type of critical incident reports, including feeding errors. DISCUSSION: Our double-blinded, randomized, controlled trial aims to examine whether fresh human milk can improve infant outcomes. The results of this study will impact both Chinese and international medical practice and feeding policy for preterm infants. In addition, data from our study will inform changes in health policy in NICUs across China, such that mothers are encouraged to enter the NICU and express fresh milk for their infants. TRIAL REGISTRATION: Chinese Clinical Trial Registry; #ChiCTR1900020577; registered January 1, 2019; http://www.chictr.org.cn/showprojen.aspx?proj=34276.

8.
Endocr Pract ; 2020 Jan 22.
Artigo em Inglês | MEDLINE | ID: mdl-31968188

RESUMO

Background: Previous studies have reported an association between iron deficiency (ID) and increased thyroid peroxidase antibody (TPO-ab) during early pregnancy. The objective of this study was to explore the relationship between ID and thyroid dysfunction, as well as thyroid autoantibodies, during the second trimester of pregnancy. Methods: A total of 1592 pregnant women (13-28 weeks gestation) were enrolled in this cross-sectional study. According to serum ferritin (SF) concentrations, they were divided into ID (SF <20 µg/L) or non-ID (SF ≥20 µg/L) groups. Logistic regression analysis was used to evaluate the association between ID and subclinical hypothyroidism (thyroid-stimulating hormone [TSH] >4.0 mIU/L and free thyroxine [FT4] within the reference range) and thyroid autoimmunity. Results: The prevalence of ID was 23.43% (373/1592). Compared with the non-ID group, the ID group had lower FT4 levels (13.94 [8.91-29.82] vs 14.63 [8.22-47.24] pmol/L, p<0.001]) and higher TSH levels (1.85 [0.01-7.84] vs 1.69 [0.01-10.2] mIU/L, p<0.05). Logistic regression analysis confirmed ID as a risk factor for increased thyroglobulin antibody (TG-ab) (odds ratio 1.974; 95% confidence interval 1.065, 3.657; p<0.05), but not for subclinical hypothyroidism or increased TPO-ab. Conclusions: ID is associated with increased TG-ab during the second trimester of pregnancy.

9.
Endocr Pract ; 2020 Jan 22.
Artigo em Inglês | MEDLINE | ID: mdl-31968197

RESUMO

Objective: Type 2 diabetes mellitus (T2DM) is a risk factor for nonalcoholic fatty liver disease (NAFLD). The aim of this study was to investigate the effect of T2DM on nonalcoholic steatohepatitis (NASH) and advanced fibrosis. Methods: A total of 221NAFLD patients who had undergone a liver biopsy were included in this study. Subjects were divided into a non-T2DM group and a T2DM group based on glycemic control. NASH was diagnosed by the joint presence of steatosis, ballooning and lobular inflammation. The steatosis, activity and fibrosis score (SAF) and NAFLD activity score (NAS) were used to evaluate the severity of NAFLD. The severity of liver fibrosis was evaluated based on the fibrosis stage. Results: The total percentagesof NASH and advanced fibrosis in this study were 95.0% and 50.2%, respectively. The percentages of NASH and advanced fibrosis in NAFLD patients with T2DM were 96.1% and56.5%, respectively, which were higher than those in the non-T2DM group. SAF (especially activity and fibrosis stage) and NAS (especially ballooning) were higher in NAFLD patients with T2DM than in NAFLD patients without T2DM. Glycemic control and insulin resistance were positively associated with SAF, NAS, and fibrosis stage. Additionally, T2DM elevated the risk of a high NAS and advanced fibrosis. Conclusion: T2DM increases the risk of serious NASH and advanced fibrosis in patients with NAFLD. Liver biopsy can be performed in NAFLD patients with T2DM to confirm the stage of NAFLD. Screening of NASH and advanced fibrosis in NAFLD patients with T2DM is needed.

10.
Food Chem ; 310: 125854, 2020 Apr 25.
Artigo em Inglês | MEDLINE | ID: mdl-31784067

RESUMO

Zeaxanthin dipalmitate (ZDP) is a major non-saponified carotenoid in fully ripe fruits of Lycium barbarum L. In the present study, response surface methodology was used to optimize the ultrasonic-assisted extraction (UAE) conditions of carotenoids from the fruits of L. barbarum, and the optimal extraction conditions were determined as follows: ultrasonic power of 360 W, ultrasonic time of 40 min and the ratio of extraction solvent to sample of 30 mL/g. An actual value of ZDP content of 5.40 mg/g and short extraction time indicated the efficiency of UAE. Furthermore, a promising high-speed counter-current chromatography (HSCCC) method was established for the purification of ZDP from the fruits of L. barbarum. With a developed two-phase solvent system composed of n-hexane/dichloromethane/acetonitrile (10/3/7, v/v/v), ZDP with a purity of higher than 95% was successfully isolated from the crude extract. This is the first report on the purification of ZDP by using HSCCC.

11.
J Exp Bot ; 71(3): 919-933, 2020 Jan 23.
Artigo em Inglês | MEDLINE | ID: mdl-31641755

RESUMO

The molecular mechanisms underlying seed dormancy and germination are not fully understood. Here, we show that Arabidopsis thaliana SEED DORMANCY 4-LIKE (AtSdr4L) is a novel specific regulator of dormancy and germination. AtSdr4L encodes a protein with an unknown biochemical function that is localized in the nucleus and is expressed specifically in seeds. Loss of function of AtSdr4L results in increased seed dormancy. The germination of freshly harvested seeds of the Atsdr4l mutant is insensitive to gibberellin (GA). After-ripened mutant seeds are hypersensitive to the GA biosynthesis-inhibitor paclobutrazol but show unaltered sensitivity to abscisic acid. Several GA biosynthesis genes and GA-regulated cell wall remodeling genes are down-regulated in the mutant in both dormant and after-ripened seeds. These results suggest that the Atsdr4l mutation causes both decreased GA biosynthesis and reduced responses. In addition, a genetic analysis indicated that AtSdr4L is epistatic to DELAY OF GERMINATION1 (DOG1) for dormancy and acts upstream of RGA-LIKE 2 (RGL2) in the GA pathway. We propose that AtSdr4L regulates seed dormancy and germination by mediating both the DOG1 and GA pathways.

12.
Phytopathology ; 110(1): 187-193, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31516080

RESUMO

Potyviral helper component protease (HC-Pro), as a major determinant of symptom expression in susceptible plants, is a likely target candidate in the production of attenuated strains for cross-protection. In this study, single or double mutations of Lys (K) to Glu (E) in the Lys-Ile-Thr-Cys motif and Arg (R) to Ile (I) in the Phe-Arg-Asn-Lys motif of the HC-Pro from the severe papaya leaf distortion mosaic virus strain DF (PLDMV-DF) reduced symptom expression and virus accumulation in infected papaya (Carica papaya) plants. The papaya plants infected with the attenuated double mutant of PLDMV-EI presented as symptomless. PLDMV-EI provided effective protection against PLDMV-DF infection in three papaya cultivars and had no effect on plant growth and development. Our result showed that PLDMV-EI is a promising mild strain for the practical use of cross-protection in the field.


Assuntos
Motivos de Aminoácidos , Carica , Peptídeo Hidrolases , Potyvirus , Motivos de Aminoácidos/genética , Carica/virologia , Mutação/genética , Peptídeo Hidrolases/genética , Potyvirus/enzimologia , Potyvirus/genética
13.
Int J Radiat Oncol Biol Phys ; 106(2): 282-290, 2020 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-31669564

RESUMO

PURPOSE: Our purpose was to compare toxicity and biochemical control in postprostatectomy patients treated with conventional (66 Gy) or dose-intensified (72 Gy) radiation therapy. METHODS AND MATERIALS: Patients who had stage pT3-4, positive surgical margins, or rising prostate-specific antigen ≥ 0.2 ng/mL after radical prostatectomy were randomly assigned to receive either 66 Gy in 33 fractions or 72 Gy in 36 fractions. A primary endpoint was to assess the difference in biochemical progression-free survival (bPFS) between these 2 cohorts, and secondary endpoints were to assess differences in genitourinary (GU), gastrointestinal (GI), and hematologic toxicities between these 2 cohorts. bPFS was estimated by the Kaplan-Meier method and toxicities were compared using the χ2 test. RESULTS: Between September 2011 and November 2016, 144 patients were enrolled: 71 patients to the 66 Gy cohort and 73 patients to the 72 Gy cohort. The median follow-up time was 48.5 months (range, 14-79 months). There was no difference in 4-year bPFS between the 66 Gy and 72 Gy cohorts (75.9% vs 82.6%; P = .299). However, in patients with a higher Gleason score (8-10), the 72 Gy cohort had statistically significant improvement in bPFS compared with the 66 Gy cohort (79.7% vs 55.7%; P = .049). Toxicity analysis showed no difference in ≥2 acute or late GI or GU toxicities between these 2 cohorts. A total of 48 patients were scored as urinary incontinence before radiation therapy, of which 39 (81.3%) reported incontinence recovery or stable at 1-year follow-up, and only 9 (18.8%) patients reported worsening. There was no difference between the 2 cohorts in urinary incontinence either at baseline or at 1-year follow-up. CONCLUSIONS: Dose escalation (72 Gy) demonstrated no improvement in 4-year bPFS compared with the 66 Gy regimen. However, the dose escalation was not associated with greater acute or late GU or GI toxicities and did not increase urinary incontinence.

14.
Funct Integr Genomics ; 20(1): 103-115, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31392586

RESUMO

Atherosclerosis, a multifactorial and chronic immune inflammatory disorder, is the main cause of multiple cardiovascular diseases. Researchers recently reported that lncRNAs may exert important functions in the progression of atherosclerosis (AS). Some studies found that lncRNAs can act as ceRNAs to communicate with each other by the competition of common miRNA response elements. However, lncRNA-associated ceRNA network in terms of atherosclerosis is limited. In present study, we pioneered to construct and systematically analyze the lncRNA-mRNA network and reveal its potential roles in carotid atherosclerotic rabbit models. Atherosclerosis was induced in rabbits (n = 3) carotid arteries via a high-fat diet and balloon injury, while age-matched rabbits (n = 3) were treated with normal chow as controls. RNA-seq analysis was conducted on rabbits carotid arteries (n = 6) with or without plaque formation. Based on the ceRNA mechanism, a ternary interaction network including lncRNA, mRNA, and miRNA was generated and an AS-related lncRNA-mRNA network (ASLMN) was extracted. Furthermore, we analyzed the properties of ASLMN and discovered that six lncRNAs (MSTRG.10603.16, 5258.4, 12799.3, 5352.1, 12022.1, and 12250.4) were highly related to AS through topological analysis. GO and KEGG enrichment analysis indicated that lncRNA MSTRG.5258.4 may downregulate inducible co-stimulator to perform a downregulated role in AS through T cell receptor signaling pathway and downregulate THBS1 to conduct a upregulated function in AS through ECM-receptor interaction pathway. Finally, our results elucidated the important function of lncRNAs in the origination and progression of AS. We provided an ASLMN of atherosclerosis development in carotid arteries of rabbits and probable targets which may lay the foundation for future research of clinical applications.

15.
Artigo em Inglês | MEDLINE | ID: mdl-31824417

RESUMO

Background and aim: Granulocyte-colony-stimulating factor (G-CSF) is highly beneficial as a general treatment for anti-thyroid drug (ATD)-induced agranulocytosis. This meta-analysis aimed to assess the clinical effects of G-CSF and non-G-CSF on recovery duration in patients with ATD-induced agranulocytosis by analyzing the overall clinical outcomes. Methods: The PubMed, Embase, Ovid, Cochrane, Google Scholar, China National Knowledge Infrastructure (CNKI) databases were searched for published studies from 1900 to 2018. No language restriction was implemented. Results: This meta-analysis included 10 published retrospective studies and one prospective study. Data were obtained from 11 trials (474 patients: 247 with G-CSF and 227 with non-G-CSF treatment). Compared with the non-G-CSF group, the G-CSF group presented shorter recovery duration [weighted mean difference (WMD) = -3.04 days, 95% confidence interval (95% CI): -4.38 to -1.69 (Z = 4.43 P = 0.000)]. However, the recovery duration varied across regions and recovery criteria. Asian patients achieved significant clinical outcomes [WMD = -3.16 days (95% CI: -4.58 to -1.74, P = 0.000)] compared with European and South American patients [WMD = -2.19 days (95% CI: -7.38 to 3.01, P = 0.409)]. Also, according to various recovery criteria, a duration of granulocyte count increase of more than 1.5 or 1.0 × 109/L [WMD = -3.50 days (95% CI: -4.82 to -2.18, P = 0.000)] revealed a better treatment effect. Conclusion: G-CSF can significantly shorten the recovery duration in patients with ATD-induced agranulocytosis.

16.
Funct Plant Biol ; 2019 Dec 09.
Artigo em Inglês | MEDLINE | ID: mdl-31813413

RESUMO

In this study we analysed the expression patterns of TaLr35PR2 and confirmed its role in Lr35-mediated adult resistance to leaf rust fungus. ß-1,3-glucanase, a pathogenesis-related protein, has a critical function in plant defence response against fungal pathogens. We previously described the full-length gene TaLr35PR2, which encodes a protein exhibiting amino acid and structural similarity to ß-1,3-glucanase, in the wheat near-isogenic line TcLr35 (GenBank accession number DQ294235.1). This work aimed to further assess TaLr35PR2 expression patterns and function in Lr35-mediated adult resistance to Puccinia triticina. Immunoblot was performed to demonstrate that TaLr35PR2 expression was triggered early by P. triticina, with expression levels markedly elevated in incompatible interaction compared with those in compatible one. Additionally, TaLr35PR2 accumulation steadily increased and overtly peaked after challenge with P. triticina through the various developmental stages of TcLr35 wheat, and remaining at similar levels after mock inoculation. Furthermore, TaLr35PR2 expression was significantly reduced in barley stripe mosaic virus (BSMV)-induced gene knockdown plants, in which pathological assessment revealed that TaLr35PR2-silenced plants was obviously susceptible to leaf rust fungus compared with wild-type TcLr35, indicating that Lr35-mediated resistance to leaf rust was diminished. These findings strongly suggest that TaLr35PR2 is involved in Lr35-mediated wheat defence against the leaf rust pathogen.

17.
J Virol Methods ; : 113795, 2019 Dec 03.
Artigo em Inglês | MEDLINE | ID: mdl-31809783

RESUMO

Two reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assays were developed for the detection of areca palm necrotic ringspot virus (ANRSV) and areca palm necrotic spindle-spot virus (ANSSV), respectively. These two emerging viruses both induce necrotic symptoms in areca palms. The coat protein (CP) gene of ANRSV and the 9 K gene of ANSSV were used to design the respective RT-LAMP primers for the assays. Each set of four primers designed for each of these viruses was found to be highly specific in the detection of the respective targeted virus. The optimal incubation conditions for the RT-LAMP assays were 63 °C for 40 min for ANRSV and at 61 °C for 40 min for ANSSV. The sensitivity of the RT-LAMP method for each of these viruses was 10-fold greater than that of the corresponding conventional reverse-transcription polymerase chain reaction (RT-PCR). The RT-LAMP assays may be useful for the rapid early detection of ANSSV and ANRSV in commercial areca palm production.

18.
Rapid Commun Mass Spectrom ; : e8715, 2019 Dec 30.
Artigo em Inglês | MEDLINE | ID: mdl-31886926

RESUMO

RATIONALE: Macleaya microcarpa (Maxim.) Fedde is a genus of Macleaya belonging to papaveraceae family. Benzylisoquinoline alkaloids are considered the main bioactive constituents of M. microcarpa. METHODS: Using HPLC-Q/TOF-MS/MS we identified the benzylisoquinoline alkaloids in the aerial parts of M. microcarpa at early flowering stage. Target profiling and identification of benzylisoquinoline alkloids in the extracted samples from fresh aerial parts of M. microcarpa were exclusively based on a personal accurate mass database of known compounds combined with the mass spectral fragmentation behavior of Macleaya alkaloids. RESULTS: Ninety-seven alkaloids including seven benzyltetrahydroisoquinolines, one aporphine, nine tetraprotoberberines, three protoberberines, two N-methyltetrahydroprotoberberines, four protopines, forty-seven dihydrobenzophenanthridines, and twenty-four benzophenanthridines were identified from the fresh aerial parts of M. microcarpa, and seventy-seven of them were detected for the first time in M. microcarpa. In addition, some of the screened alkaloids were related to the biosynthetic pathways of sanguinarine and chelerythrine. CONCLUSIONS: The integrated method is sensitive and reliable for screening and identifying trace or ultra trace isoquinoline alkaloids and contributed to a better understanding of benzylisoquinoline alkaloids in fresh aerial parts of M. microcarpa.

19.
Hua Xi Kou Qiang Yi Xue Za Zhi ; 37(6): 577-582, 2019 Dec 01.
Artigo em Chinês | MEDLINE | ID: mdl-31875433

RESUMO

OBJECTIVE: To observe the effect of protein kinase D1 (PKD1) on the growth and metabolism of oral squamous cell carcinoma HSC-4 cells and related molecular mechanisms in the tumor microenvironment. METHODS: HSC-4 cell lines were transfected with shRNA plasmids. Three groups (Wild, control-shRNA, and PKD1-shRNA) were cultured under acidic or hypoxic environment for a certain time. Western blot was used to detect the expression of autophagy-related and glycolytic-related proteins. The proliferation changes were detected by CCK-8 kits. RESULTS: The PKD1-knockdown HSC-4 cell line was established. PKD1 silencing increased autophagy activity. Under hypoxic and acidic conditions, the PKD1-knockdown HSC-4 cells showed lower proliferation than the parental cells. PKD1-knockdown also decreased the expression of hypoxia induciblefactor 1α (HIF-1α) and pyruvate kinase M2 (PKM2). CONCLUSIONS: Under hypoxic and acidic conditions, PKD1 gene silencing can increase apoptotic autophagy activity. Downregulated PKD1 gene expression can reduce the glycolysis of oral squamous cell carcinoma cells and inhibit tumor cell proliferation. This study revealed the important role of PKD1 in the metabolism and growth of oral squamous cell carcinoma, making it a possible target for the treatment of oral squamous cell carcinoma.


Assuntos
Carcinoma de Células Escamosas , Neoplasias Bucais , Linhagem Celular Tumoral , Proliferação de Células , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia , Proteínas Quinases , Microambiente Tumoral
20.
Hua Xi Kou Qiang Yi Xue Za Zhi ; 37(6): 583-588, 2019 Dec 01.
Artigo em Chinês | MEDLINE | ID: mdl-31875434

RESUMO

OBJECTIVE: This study aimed to investigate the role of protein kinase D (PKD)1 in regulating the growth, apop-tosis, and drug sensitivity of the squamous carcinoma cell line SCC-25. METHODS: The SCC-25 cell line was transfected with either the control-shRNA or PKD1-shRNA plasmids. The stable transfected cells were selected, and the efficiency of PKD1 knockdown was detected by Western blot. The growth and apoptosis of SCC-25 were analyzed with a cell counting kit-8 (CCK8) and flow cytometry. The 50% inhibitory concentrations (IC50) of paclitaxel in the control and PKD1 knockdown cell lines were detected by CCK-8. The expression levels of Bax, Bcl-2, and P-gp were detected by Western blot. RESULTS: PKD1 was constitutively expressed and phosphorylated in various cancer cell lines. Inhibiting the expression of PKD1 in SCC-25 cells by RNA interference could inhibit the growth and promote the apoptosis of SCC-25 cells via downregulating Bcl-2 expression. Additionally, inhibiting PKD1 expression could downregulate the expression of P-gp, thereby decreasing both the IC50 and resistance index of paclitaxel. CONCLUSIONS: PKD1 plays an important role in regulating the biobehavior of SCC-25. It is a potential therapeutic target for oral squamous carcinoma.


Assuntos
Carcinoma de Células Escamosas , Neoplasias Bucais , Apoptose , Linhagem Celular Tumoral , Proliferação de Células , Humanos
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