Plant virus-based expression systems are an alternative expression platform for the production of clinically and industrially useful recombinant proteins. Nonetheless, due to a lack of viral vector ...with the commercial potentials, it is urgent to design and develop new, versatile, and efficient plant virus vectors. The genome of
Tomato bushy stunt virus
(TBSV) offers an attractive alternative to being modified as a vector for producing heterologous proteins in plants. Here, we developed a set of novel fusion and non-fusion TBSV-CP replacement vectors, which provide more flexible and efficient tools for expressing proteins of interest in plants. An alternative tobacco plant,
Nicotiana excelsiana
, was used in this study as a host for newly constructed TBSV vectors because the unwanted necrotic effects were reported on the commonly used
Nicotiana benthamiana
host associated with expression of TBSV-encoded P19 protein. The data showed that TBSV vectors caused a symptomless infection and overexpressed reporter gene in
N. excelsiana
leaves, demonstrating that
N. excelsiana
is an ideal host plant for TBSV-mediated heterologous gene expression
.
Moreover, a TBSV non-fusion vector, dAUG, shows the similar accumulation level of reporter proteins to that of TMV- and PVX-based vectors in side-by-side comparison and provides more flexible aspects than the previously developed TBSV vectors. Collectively, our newly developed TBSV expression system adds a new member to the family of plant viral expression vectors and meanwhile offers a flexible and highly effective approach for producing proteins of interest in plants.
Key points
•
The TBSV-based transient expression system has been significantly improved.
•
The necrotic effects caused by viral P19 protein were avoided by the usage of N. excelsiana as a host plant.
•
The expression level of the non-fusion vector was similar to the most effective virus vectors reported so far.
Toll-like receptor 4 (TLR4) is an essential sensor related to tumorigenesis, and overexpression of TLR4 in human tumors often correlates with poor prognosis. Atractylenolide-I (AT-I), a novel ...TLR4-antagonizing agent, is a major bioactive component from
. Emerging evidence suggests that AT-I exerts anti-tumor effects on various cancers such as colorectal cancer, bladder cancer and melanoma. Nevertheless, the effects of AT-I on mammary tumorigenesis remain unclear.
In order to ascertain the correlation of TLR4/NF-κB pathway with breast cancer, the expression of TLR4 and NF-κB in normal breast tissues and cancer tissues with different TNM-stages was detected by human tissue microarray and immunohistochemistry technology. The effects of AT-I on tumorigenesis were investigated by cell viability, colony formation, apoptosis, migration and invasion assays in two breast cancer cells (MCF-7 and MDA-MB-231), and N-Nitroso-N-methylurea induced rat breast cancer models were developed to evaluate the anti-tumor effects of AT-I
. The possible underlying mechanisms were further explored by western blot and ELISA assays after a series of LPS treatment and TLR4 knockdown experiments.
We found that TLR4 and NF-κB were significantly up-regulated in breast cancer tissues, and was correlated with advanced TNM-stages. AT-I could inhibit TLR4 mediated NF-κB signaling pathway and decrease NF-κB-regulated cytokines in breast cancer cells, thus inhibiting cell proliferation, migration and invasion, and inducing apoptosis of breast cancer cells. Furthermore, AT-I could inhibit N-Nitroso-N-methylurea-induced rat mammary tumor progression through TLR4/NF-κB pathway.
Our findings demonstrated that TLR4 and NF-κB were over expressed in breast cancer, and AT-I could suppress tumorigenesis of breast cancer via inhibiting TLR4-mediated NF-κB signaling pathway.
In hidden Markov chain (HMC) models, widely used for target tracking, the process noise and measurement noise are in general assumed to be independent and Gaussian for mathematical simplicity. ...However, the independence and Gaussian assumptions do not always hold in practice. For instance, in a typical radar tracking application, the measurement noise is correlated over time as the sampling frequency of a radar is generally much higher than the bandwidth of the measurement noise. In addition, target maneuvers and measurement outliers imply that the process noise and measurement noise are non-Gaussian. To solve this problem, we resort to triplet Markov chain (TMC) models to describe stochastic systems with correlated noise and derive a new filter under the maximum correntropy criterion to deal with non-Gaussian noise. By stacking the state vector, measurement vector, and auxiliary vector into a triplet state vector, the TMC model can capture the complete dynamics of stochastic systems, which may be subjected to potential parameter uncertainty, non-stationarity, or error sources. Correntropy is used to measure the similarity of two random variables; unlike the commonly used minimum mean square error criterion, which uses only second-order statistics, correntropy uses second-order and higher-order information, and is more suitable for systems in the presence of non-Gaussian noise, particularly some heavy-tailed noise disturbances. Furthermore, to reduce the influence of round-off errors, a square-root implementation of the new filter is provided using QR decomposition. Instead of the full covariance matrices, corresponding Cholesky factors are recursively calculated in the square-root filtering algorithm. This is more numerically stable for ill-conditioned problems compared to the conventional filter. Finally, the effectiveness of the proposed algorithms is illustrated via three numerical examples.
Root rot, caused by several pathogens, is one of the most devastating diseases on
Astragalus membranaceus
, and can lead to serious yield loss.
Fusarium acuminatum
and
F. solani
are known to be major ...pathogens causing root rot of
A. membranaceus
in Shanxi Province, China. It is well-known that accurate identification of the pathogen and early diagnosis of the symptoms play a significantly important role to control this disease in the early or mid-term stage. Here, two new loop-mediated isothermal amplification (LAMP) assays were developed to detect
F. acuminatum
and
F. solani
based on the partial translation elongation factor-1α (
TEF-1α
) gene region. The LAMP products were successfully assessed by visual assessment using SYBR Green I, as well as electrophoresis on 2.0% agarose gel. The LAMP reactions for
F. acuminatum
and
F. solani
were performed under the optimized conditions of 63 °C and 62 °C for 60 min, respectively. The detection limit of LAMP assay was 100 fg/μL for
F. acuminatum
and 1 pg/μL for
F. solani
. Additionally, the two LAMP assays are highly specific to target
Fusarium
species, and could differentiate
F. acuminatum
and
F. solani
from other
Fusarium
species. In the field, the suspected diseased samples were used to verify the feasibility of the developed assays. As a result, the two rapid and specific LAMP assays could be applied for direct detection of
F. acuminatum
and
F. solani
on
A. membranaceus
, and precise diagnosis of
A. membranaceus
root rot caused by
F. acuminatum
and
F. solani
.
Codeine, a prodrug used as an opioid agonist, is metabolized to the active product morphine by CYP2D6. This study aimed to establish physiologically based pharmacokinetic (PBPK) models of codeine and ...morphine and explore the influence of CYP2D6 genetic polymorphisms on the pharmacokinetics of codeine and morphine.
An initial PBPK modeling of codeine in healthy adults was established using PK-Sim
software and subsequently extrapolated to CYP2D6 phenotype-related PBPK modeling based on the turnover frequency (K
) of CYP2D6 for different phenotype populations (UM, EM, IM, and PM). The mean fold error (MFE) and geometric mean fold error (GMFE) methods were used to compare the differences between the predicted and observed values of the pharmacokinetic parameters to evaluate the accuracy of PBPK modeling. The validated models were then used to support dose safety for different CYP2D6 phenotypes.
The developed and validated CYP2D6 phenotype-related PBPK model successfully predicted codeine and morphine dispositions in different CYP2D6 phenotypes. Compared with EMs, the predicted AUC
value of morphine was 98.6% lower in PMs, 60.84% lower in IMs, and 73.43% higher in UMs. Morphine plasma exposure in IMs administered 80 mg of codeine was roughly comparable to that in EMs administered 30 mg of codeine. CYP2D6 UMs may start dose titration to achieve an optimal individual regimen and avoid a single dose of over 20 mg. Codeine should not be used in PMs for pain relief, considering its insufficient efficacy.
PBPK modeling can be applied to explore the dosing safety of codeine and can be helpful in predicting the effect of CYP2D6 genetic polymorphisms on drug-drug interactions (DDIs) with codeine in the future.
Display omitted
•Nrf2-MRP2/4 pathway is essential for Tan IIA to prevent APAP-induced hepatotoxicity.•Nrf2 directly regulates MRP2/4 via binding to AREs in their promoters.•HOTAIR performs an ...epigenetic suppression on Nrf2-MRP2/4 pathway.•Tan IIA activates Nrf2-MRP2/4 pathway via downregulating HOTAIR.
Tanshinone IIA (Tan IIA), an active component in S. miltiorrhiza, has been reported to have excellent antioxidant and detoxifying activity. Here, we prove that Tan IIA attenuates acetaminophen-induced hepatotoxicity from a pharmacokinetic perspective. Compared with acetaminophen (APAP, 200 mg/kg) treated mice, Tan IIA pretreatment (30 mg/kg/d) not only reduced the plasma level of the toxic metabolite N-acetyl-p-benzoquinone imine (NAPQI) but also increased its bile level. After Tan IIA pretreatment, significant induction of nuclear factor E2-related factor 2 (Nrf2), multidrug resistance-associated protein 2 (Mrp2), and multidrug resistance-associated protein 4 (Mrp4) mRNA and protein expression was detected in Nrf2+/+ mouse liver, however, much lower increase of Mrp2 and Mrp4 mRNA and protein expression was observed in Nrf2−/− mouse liver. Luciferase reporter and chromatin immunoprecipitation assays demonstrated that Nrf2 bounds to antioxidant responsive elements (AREs) of the MRP2 and MRP4 promoter, thus regulating the expression of MRP2 and MRP4. in vitro experiments revealed that Tan IIA increase Nrf2, MRP2, and MRP4 expression through a mechanism of inhibiting the expression of HOX transcript antisense RNA (HOTAIR) which belongs to long non-coding RNAs. Collectively, the present results demonstrated that Tan IIA could protect against APAP-induced hepatotoxicity by altering the pharmacokinetic characteristics of APAP and its metabolites via HOTAIR-Nrf2-MRP2/4 signaling pathway, and HOTAIR plays a pivotal role in the MRP2 and MRP4 expression regulated by Nrf2.
To systematically investigate the effects of two methods used for laser-assisted hatching (LAH) on clinical outcomes after day 4 (D4) on frozen-embryo-transfer (FET) cycles. Data from 11471 infertile ...patients who underwent FET cycles between January 2014 and October 2018 was retrospectively analyzed. The 1410 patients who met the inclusion criteria were further categorized into two groups based on the hatching procedure used: the thinning laser-assisted hatching group (T-LAH, 716 patients), and the drilling laser-assisted hatching group (D-LAH, 694 patients). The baseline characteristics of the patients were consistent between the two groups. However, the rates of implantation and clinical pregnancy were significantly higher in the T-LAH group compared to the D-LAH group (32.73% vs. 29.09%,
P
< 0.01, and 50.98% vs. 43.95%,
P
< 0.01). The proportion of live birth was also higher in the T-LAH group, but the difference was insignificant (39.11% vs. 36.89%,
P
> 0.05). Moreover, there were no significant differences in rates of miscarriages, multiple pregnancies, ectopic pregnancies, preterm births, and congenital disabilities between the two groups. Nonetheless, significantly higher rates of implantation and pregnancy were reported in the T-LAH group compared to the D-LAH group among patients aged <35 years, patients with at least one previously failed cycle, and patients with an endometrial thickness of 8–10 mm. T-LAH is superior to D-LAH in improving clinical implantation and pregnancy outcomes in D4 FET, particularly in patients aged <35 years with at least one previously failed cycle or an endometrial thickness of 8–10 mm. The findings of this study provide theoretical support for clinical individualized diagnosis and treatment of patients with infertility.
Cardiovascular disease (CVD) is a serious public health risk, and prevention and treatment efforts are urgently needed. Effective preventive and therapeutic programs for cardiovascular disease are ...still lacking, as the causes of CVD are varied and may be the result of a multifactorial combination. Mitophagy is a form of cell-selective autophagy, and there is increasing evidence that mitophagy is involved in cardioprotective processes. Recently, many studies have shown that FUN14 domain-containing protein 1 (FUNDC1) levels and phosphorylation status are highly associated with many diseases, including heart disease. Here, we review the structure and functions of FUNDC1 and the path-ways of its mediated mitophagy, and show that mitophagy can be effectively activated by dephosphorylation of Ser13 and Tyr18 sites, phosphorylation of Ser17 site and ubiquitination of Lys119 site in FUNDC1. By effectively activating or inhibiting excessive mitophagy, the quality of mitochondria can be effectively controlled. The main reason is that, on the one hand, improper clearance of mitochondria and accumulation of damaged mitochondria are avoided, and on the other hand, excessive mitophagy causing apoptosis is avoided, both serving to protect the heart. In addition, we explore the possible mechanisms by which FUNDC1-mediated mitophagy is involved in exercise preconditioning (EP) for cardioprotection. Finally, we also point out unresolved issues in FUNDC1 and its mediated mitophagy and give directions where further research may be needed.
To compare cumulative live birth rate (LBR) between progestin-primed ovarian stimulation (PPOS) and GnRH antagonist protocols of preimplantation genetic testing (PGT) cycles in different populations.
...This was a retrospective cohort study. A total of 865 patients were enrolled and separate analyses were performed for three populations: 498 patients with predicted normal ovarian response (NOR), 285 patients with PCOS, and 82 patients with predicted poor ovarian response (POR). The primary outcome was cumulative LBR for one oocyte retrieval cycle. The results of response to ovarian stimulation were also investigated, including numbers of oocytes retrieved, MII oocytes, 2PN, blastocysts, good-quality blastocysts, and usable blastocysts after biopsy, as well as rates of oocyte yield, blastocyst formation, good-quality blastocysts, and moderate or severe OHSS. Univariable and multivariable logistic regression analyses were used to identify potential confounders that may be independently associated with cumulative live birth.
In NOR, the cumulative LBR of PPOS protocol was significantly lower than that of GnRH antagonists (28.4% vs. 40.7%;
=0.004). In multivariable analysis, the PPOS protocol was negatively associated with cumulative LBR (adjusted OR=0.556; 95% CI, 0.377-0.822) compared to GnRH antagonists after adjusting for potential confounders. The number and ratio of good-quality blastocysts were significantly reduced in PPOS protocol compared to GnRH antagonists (2.82 ± 2.83 vs. 3.20 ± 2.79;
=0.032 and 63.9% vs. 68.5%;
=0.021), while numbers of oocytes, MII oocytes and 2PN did not show any significant difference between GnRH antagonist and PPOS protocols. PCOS patients had similar outcomes as NOR. The cumulative LBR of PPOS group appeared to be lower than that of GnRH antagonists (37.4% vs. 46.1%;
=0.151), but not significantly. Meanwhile, the proportion of good-quality blastocysts in PPOS protocol was also lower compared to GnRH antagonists (63.5% vs. 68.9%;
=0.014). In patients with POR, the cumulative LBR of PPOS protocol was comparable to that of GnRH antagonists (19.2% vs. 16.7%;
=0.772). There was no statistical difference in the number and rate of good-quality blastocysts between the two protocols in POR, while the proportion of good-quality blastocysts appeared to be higher in PPOS group compared to GnRH antagonists (66.7% vs. 56.3%;
=0.182). In addition, the number of usable blastocysts after biopsy was comparable between the two protocols in three populations.
The cumulative LBR of PPOS protocol in PGT cycles is lower than that of GnRH antagonists in NOR. In patients with PCOS, the cumulative LBR of PPOS protocol appears to be lower than that of GnRH antagonists, albeit lacking statistical difference, whereas in patients with diminished ovarian reserve, the two protocols were comparable. Our findings suggest the need for caution when choosing PPOS protocol to achieve live births, especially for normal and high ovarian responders.
Does aquaporin 3 (AQP3) affect the migration and invasion of human extravillous trophoblast (HTR8/Svneo) cells?
A lentivirus infection system was used to construct stable cell lines with either AQP3 ...knockdown or overexpression. RT-PCR and western blotting were used to verify the efficiencies of AQP3 knockdown or overexpression in HTR8/Svneo cells at mRNA and protein levels, respectively. Cell Counting Kit-8 and flow cytometry assays were used to detect the influence of AQP3 knockdown or overexpression on proliferation and apoptosis of HTR8/Svneo cells. In addition, wound healing and Transwell invasion assays were used to detect the effects of AQP3 knockdown or overexpression on migration and invasion capabilities of HTR8/Svneo cells. An Agilent gene chip was used to screen for significant differentially expressed genes after AQP3 knockdown. Finally, mechanisms by which AQP3 influences the migration and invasion of HTR8/Svneo cells were explored using bioinformatic analysis.
Compared with controls, migration and invasion capabilities of HTR8/Svneo cells were significantly reduced after AQP3 knockdown, and significantly increased after AQP3 overexpression. Subsequent bioinformatic analysis of gene chip expression profiles indicated downregulation of genes related to adhesion such as PDGF-B, as well as signaling pathways (such as PIK3/AKT, NF-κB, and TNF) after AQP3 knockdown.
AQP3 could significantly promote migration and invasion capabilities of human extravillous trophoblasts, it may mediate embryo invasion and adhesion to endometrium by regulating PDGF-B, PIK3/AKT signaling pathways, although this requires further verification.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
PDF
