Expression and Function of VEGFRs in Normal Epidermis and Psoriatic Lesional Keratinocytes.

The study aimed to explore the expression and function of VEGFRs in normal epidermis and keratinocytes of psoriatic lesions. In this study, the expression and role of VEGFRs in keratinocytes were examined using examples from psoriatic and healthy individuals. The experiment was completed by immunofluorescence analysis, reverse transcription polymerase chain reaction, Western blot, and real-time quantitative RT-PCR after the skin of nonlesional, adjacent, and lesional skin was excised. Observations indicated that in non-lesional psoriatic areas and adjacent lesional areas of the skin of psoriasis patients, the fluorescent signals of VEGFR-1 and VEGFR-2 were strongly labelled with keratinocytes, and in psoriatic lesions, keratinocytes were present throughout the entire thickness of the epidermis, with the exception of the stratum corneum. The distribution of VEGFR-3 in psoriatic nonlesional and adjacent lesional skin was consistent with that in normal epidermis, whereas all layers of the epidermis of psoriatic lesions expressed VEGFR-3. The mRNA expression levels of VEGFR-1,2,3 steadily increased from the normal epidermis to the psoriatic nonlesional, adjacent lesional, and perilesional areas, with the lesional epidermis' keratinocytes exhibiting the greatest levels of mRNA expression. Ca ions upregulate VEGFR-1,2,3 mRNA and protein expression in keratinocytes of nonlesional areas of psoriasis. VEGFRs protein expression and cortical IOD values of psoriatic and normal population cells showed a positive correlation. Hence, in comparison to normal epidermal keratinocytes, psoriatic lesional regions' keratinocytes considerably enhanced their expression of VEGFR-1,2,3 mRNA and protein. The overexpression of VEGFR-1,2,3 in psoriatic lesions may be encouraged by VEGF and Ca þ ions.

Preparation of Dihydromyricetin-Loaded Self-Emulsifying Drug Delivery System and Its Anti-Alcoholism Effect.

Intraperitoneal injection of dihydromyricetin (DMY) has shown promising potential in the treatment of alcoholism. However, its therapeutic effect is limited due to its low solubility, poor stability, and high gut-liver first-pass metabolism, resulting in very low oral bioavailability. In this study, we developed a DMY-loaded self-emulsifying drug delivery system (DMY-SEDDS) to enhance the oral bioavailability and anti-alcoholism effect of DMY. DMY-SEDDS improved the oral absorption of DMY by facilitating lymphatic transport. The area under the concentration-time curve (AUC) of DMY in the DMY-SEDDS group was 4.13-fold higher than in the DMY suspension group. Furthermore, treatment with DMY-SEDDS significantly enhanced the activities of alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) in the liver of mice (p < 0.05). Interestingly, DMY-SEDDS also increased ADH activity in the stomach of mice with alcoholism (p < 0.01), thereby enhancing ethanol metabolism in the gastrointestinal tract and reducing ethanol absorption into the bloodstream. As a result, the blood alcohol concentration of mice with alcoholism was significantly decreased after DMY-SEDDS treatment (p < 0.01). In the acute alcoholism mice model, compared to saline treatment, DMY-SEDDS prolonged the onset of LORR (loss of righting reflex) (p < 0.05) and significantly shortened the duration of LORR (p < 0.01). Additionally, DMY-SEDDS treatment significantly reduced gastric injury in acute alcoholism mice. Collectively, these findings demonstrate the potential of DMY-SEDDS as a treatment in the treatment of alcoholism.

Pirh2 restricts influenza A virus replication by modulating short-chain ubiquitination of its nucleoprotein.

Influenza A viruses (IAVs) rely on viral ribonucleoprotein (vRNP) complexes to control transcription and replication. Each vRNP consists of one viral genomic RNA segment associated with multiple nucleoproteins (NP) and a trimeric IAV RNA polymerase complex. Previous studies showed that post-translational modifications of vRNP components, such as NP, by host factors would in turn affect the IAV life cycle or modulate host anti-viral response. In this study, we found host E3 ubiquitin ligase Pirh2 interacted with NP and mediated short-chain ubiquitination of NP at lysine 351, which suppressed NP-PB2 interaction and vRNP formation. In addition, we showed that knockdown of Pirh2 promoted IAV replication, whereas overexpression of Pirh2 inhibited IAV replication. However, Pirh2-ΔRING lacking E3 ligase activity failed to inhibit IAV infection. Moreover, we showed that Pirh2 had no effect on the replication of a rescued virus, WSN-K351R, carrying lysine-to-arginine substitution at residue 351. Interestingly, by analyzing human and avian IAVs from 2011 to 2020 in influenza research databases, we found that 99.18% of 26 977 human IAVs encode lysine, but 95.3% of 9956 avian IAVs encode arginine at residue 351 of NP protein. Consistently, knockdown of Pirh2 failed to promote propagation of two avian-like influenza viruses, H9N2-W1 and H9N2-C1, which naturally encode arginine at residue 351 of NP. Taken together, we demonstrated that Pirh2 is a host factor restricting IAV infection by modulating short-chain ubiquitination of NP. Meanwhile, it is noteworthy that residue 351 of NP targeted by Pirh2 may associate with the evasion of human anti-viral response against avian-like influenza viruses.

Self-Assembled Micelles Improve the Oral Bioavailability of Dihydromyricetin and Anti-Acute Alcoholism Activity.

Dihydromyricetin (DMY) is highly effective in counteracting acute alcohol intoxication. However, its poor aqueous solubility and permeability lead to the low oral bioavailability and limit its clinic application. The aim of this work is to use Solutol®HS15 (HS 15) as surfactant to develop novel micelle to enhance the oral bioavailability of DMY by improving its solubility and permeability. The DMY-loaded Solutol®HS15 micelles (DMY-Ms) were prepared by the thin-film hydration method. The particle size of DMY-Ms was 13.97 ± 0.82 nm with an acceptable polydispersity index of 0.197 ± 0.015. Upon entrapped in micelles, the solubility of DMY in water was increased more than 25-fold. The DMY-Ms had better sustained release property than that of pure DMY. In single-pass intestinal perfusion models, the absorption rate constant (Ka) and permeability coefficient (Papp) of DMY-Ms were 5.5-fold and 3.0-fold than that of pure DMY, respectively. The relative bioavailability of the DMY-Ms (AUC0-∞) was 205% compared with that of pure DMY (AUC0-∞), indicating potential for clinical application. After administering DMY-Ms, there was much lower blood alcohol level and shorter duration of the loss of righting relax (LORR) in drunk animals compared with that treated by pure DMY. In addition, the oral administration of DMY-Ms greatly reduced oxidative stress, and significantly defended liver and gastric mucosa from alcoholic damages in mice with alcohol-induced tissue injury. Taken together, HS 15-based micelle system greatly improves the bioavailability of DMY and represents a promising strategy for the management of acute alcoholism. Graphical abstract.

Predicting the Animal Susceptibility and Therapeutic Drugs to SARS-CoV-2 Based on Spike Glycoprotein Combined With ACE2.

Recently, a few animals have been frequently reported to have been diagnosed with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Whether they are SARS-CoV-2 intermediate hosts is worthy of great attention. The interaction of SARS-CoV-2 spike protein and its acceptor protein ACE2 is an important issue in determining viral host range and cross-species infection, while the binding capacity of Spike protein to ACE2 of different species is unknown. Here, we used the atomic structure model of SARS-CoV-2 and human ACE2 to assess the receptor utilization capacity of ACE2s from 10 kinds of animals. Results show that chimpanzees, domestic cats and cattles are more susceptible to infection by SARS-CoV-2. Cats in particular, such as pet cats and stray cats, interact very closely with humans, implying the necessity to carefully evaluate the risk of cats during the current COVID-19 pandemic. Furthermore, based on ACE2(cats)-SARS-CoV-2-RBD model, through high-throughput screening methods using a pool of 30,000 small molecules, eight compounds were selected for binding free energy calculations. All the eight compounds can effectively interfere with the binding of ACE2 and Spike protein, especially Nelfinavir, providing drug candidates for the treatment and prevention of SARS-CoV-2, suggesting further assessment of the anti-SARS-CoV-2 activity of these compounds in cell culture. Although we only reported the results of the simulation, and more laboratory and epidemiological investigation are required. Like cats are a risk factor, we can further detect SARS-CoV-2 according to the susceptibility of different animals, find the potential host of infection, and completely cut off the living space of the virus. Especially, cats could be a choice of animal model for screening antiviral drugs or vaccine candidates against SARS-CoV-2.

Comparative pharmacokinetic study of PEGylated gemcitabine and gemcitabine in rats by LC-MS/MS coupled with pre-column derivatization and MSALL technique.

Gemcitabine is a small molecular antitumor compound used to treat many types of solid tumors. The clinical application of gemcitabine is limited by its short biological half-life, rapid metabolism and poor tumor tissue targeting. The covalent attachment of polyethylene glycol to gemcitabine is a promising technique to overcome these limitations. After PEGylation, PEGylated gemcitabine could be metabolized into gemcitabine and its metabolites in vivo. Due to the scale effect of PEGylated gemcitabine, the DMPK process of the original drug is greatly changed. Therefore, understanding the pharmacokinetic behavior of PEGylated gemcitabine, gemcitabine and the metabolite dFdU in vivo is really important to clarify the antitumoral activity of these compounds. It would also guide the development of other PEGylated drugs. Due to the complex structure and diverse physiochemical property of PEG, direct quantification analysis of PEGylated gemcitabine presented many challenges in terms of assay sensitivity, selectivity, and robustness. In this article, a data-independent acquisition method, MSALL-based approach using electrospray ionization (ESI) coupled quadrupole time of flight mass spectrometry (MS), was utilized for the determination of PEGylated gemcitabine in rat plasma. The technique consists of a Q1 mass window through all the precursor ions, fragmenting and recording all product ions. PEGylated gemcitabine underwent dissociation in collision cell to generate a series of PEG related ions at m/z 89.0604, 133.0868, 177.1129 of 2, 3, 4 repeating ethylene oxide subunits and PEGylated gemcitabine related ions at m/z 112.0514. PEGylated gemcitabine was detected by the high resolution extracted ions based on the specific compound. For gemcitabine and dFdU, the study used derivatization of these high polarity compounds with dansyl chloride to improve their chromatographic retention. This paper describes comparative pharmacokinetic study of PEGylated gemcitabine and gemcitabine in rats by LC-MS/MS coupled with pre-column derivatization and MSALL technique. The results show that PEGylation could reduce the drug clearance of the conjugated compounds and increase the drug plasma half-life. After administration of PEGylated gemcitabine, the exposure of the free gemcitabine in vivo is lower than administration of gemcitabine, which means that PEGylated gemcitabine possesses lower toxicity compared with gemcitabine.

Parents' perceptions of diagnostic genetic testing for children with inherited retinal disease in China.

BACKGROUND: In this study, we aim to investigate the awareness of, attitudes toward, and experiences with diagnostic genetic testing among parents of children suspected of having inherited retinal disease (IRDs) in China. METHODS: Semistructured, face-to-face, and in-depth interviews were carried out with parents of children with suspected IRDs in this qualitative study. Inductive content analysis was used for data processing. RESULTS: Forty-six parents participated in our interviews, and 47.8% of them supported genetic testing for following four main reasons: to help in making informed reproductive health decisions, to prepare for novel potential treatment, to identify the underlying causes of IRDs, and to satisfy curiosity about the heredity of IRDs. Among them, 19.6% were opposed to the testing for four main reasons, namely lack of therapeutic benefit, difficulty in affording the testing cost, doubt in the accuracy of clinical diagnosis, and the presence of concerns about the limitations of genetic testing. 47.8% of the parents expressed concerns that the genetic findings might lead to potential psychological stress. CONCLUSION: In this study, we showed that nearly half of the parents supported genetic testing mainly for family planning, and a fifth of the parents were opposed to the testing mainly for lack of therapeutic benefit. Moreover, half of the parents expressed concern that a positive genetic result may create potential psychological burden to the parents and children.

Ultra-low dose lung CT perfusion regularized by a previous scan.

RATIONALE AND OBJECTIVES: A previous scan-regularized reconstruction (PSRR) method was proposed to reduce radiation dose and applied to lung perfusion studies. Normal and ultra-low-dose lung computed tomographic perfusion studies were compared in terms of the estimation accuracy of pulmonary functional parameters. MATERIALS AND METHODS: A sequence of sheep lung scans were performed in three prone, anesthetized sheep at normal and ultra-low doses. A scan protocol was developed for the ultra-low-dose studies with electrocardiographic gating: time point 1 for a normal x-ray dose scan (100 kV, 150 mAs) and time points 2 to 21 for low-dose scans (80 kV, 17 mAs). A nonlinear diffusion-based post-filtering method was applied to the difference images between the low-dose images and the high-quality reference image. The final images at 20 time points were generated by fusing the reference image with the filtered difference images. RESULTS: The power spectra of perfusion images and coherences in the normal scans showed a great improvement in image quality of the ultra-low-dose scans with PSRR relative to those without RSRR. The gamma variate fitting and the repeatability of the measurements of the mean transit time demonstrated that the key parameters of lung functions can be reliably accessed using PSRR. The variability of the ultra-low-dose scan results obtained using PSRR was not substantially different from that between two normal-dose scans. CONCLUSIONS: This study demonstrates that an approximate 90% reduction in radiation dose is achievable using PSRR without compromising quantitative computed tomographic measurements of regional lung function.

Digital tomosynthesis aided by low-resolution exact computed tomography.

Tomosynthesis reconstructs 3-dimensional images of an object from a significantly fewer number of projections as compared with that required by computed tomography (CT). A major problem with tomosynthesis is image artifacts associated with the data incompleteness. In this article, we propose a hybrid tomosynthesis approach to achieve higher image quality as compared with competing methods. In this approach, a low-resolution CT scan is followed by a high-resolution tomosynthesis scan. Then, both scans are combined to reconstruct images. To evaluate the image quality of the proposed method, we design a new breast phantom for numerical simulation and physical experiments. The results show that images obtained by our approach are clearly better than those obtained without such a CT scan.

Local ROI Reconstruction via Generalized FBP and BPF Algorithms along More Flexible Curves.

We study the local region-of-interest (ROI) reconstruction problem, also referred to as the local CT problem. Our scheme includes two steps: (a) the local truncated normal-dose projections are extended to global dataset by combining a few global low-dose projections; (b) the ROI are reconstructed by either the generalized filtered backprojection (FBP) or backprojection-filtration (BPF) algorithms. The simulation results show that both the FBP and BPF algorithms can reconstruct satisfactory results with image quality in the ROI comparable to that of the corresponding global CT reconstruction.

Exact BPF and FBP algorithms for nonstandard saddle curves.

A hot topic in cone-beam CT research is exact cone-beam reconstruction from a general scanning trajectory. Particularly, a nonstandard saddle curve attracts attention, as this construct allows the continuous periodic scanning of a volume-of-interest (VOI). Here we evaluate two algorithms for reconstruction from data collected along a nonstandard saddle curve, which are in the filtered backprojection (FBP) and backprojection filtration (BPF) formats, respectively. Both the algorithms are implemented in a chord-based coordinate system. Then, a rebinning procedure is utilized to transform the reconstructed results into the natural coordinate system. The simulation results demonstrate that the FBP algorithm produces better image quality than the BPF algorithm, while both the algorithms exhibit similar noise characteristics.

Computed tomography simulation with superquadrics.

Accurate and efficient simulation of an x-ray transform for representative structures plays an important role in research and development of x-ray CT, for the evaluation and improvement of CT image reconstruction algorithms, in particular. Superquadrics are a family of three-dimensional objects, which can be used to model a variety of anatomical structures. In this paper, we propose an algorithm for the computation of x-ray transforms for superellipsoids and tori with a monochromatic x-ray. Their usefulness is demonstrated by projection and reconstruction of a superquadric-based thorax phantom. Our work indicates that superquadric modeling provides a more realistic visualization than quadratic modeling, and a faster computation than spline methods.

A differentiable Shepp-Logan phantom and its applications in exact cone-beam CT.

Recently, several exact cone-beam reconstruction algorithms, such as the generalized filtered-backprojection (FBP) and backprojection-filtration (BPF) methods, have been developed to solve the long object problem. Although the well-known 3D Shepp-Logan phantom (SLP) is often used to validate these algorithms, it is deficient due to the discontinuity of the SLP. In this paper, we first construct a differentiable polynomial function to approximate the unit rectangular function on [-1, 1]. Then, we use this function to obtain a differentiable ellipsoid phantom, whose x-ray transform is differentiable for any smooth scanning trajectory. Finally, we propose a differentiable Shepp-Logan phantom (DSLP) for numerical simulation of the exact cone-beam CT algorithms. Our numerical simulation shows that the reconstructed DSLP has a better image quality than the reconstructed SLP, and is complementary to the traditional SLP for evaluation of the exact cone-beam CT algorithms.

A general exact reconstruction for cone-beam CT via backprojection-filtration.

In this paper, we prove a generalized backprojection-filtration formula for exact cone-beam image reconstruction with an arbitrary scanning locus. Our proof is independent of the shape of the scanning locus, as long as the object is contained in a region where there is a chord through any interior point. As special cases, this generalized formula can be applied with cone-beam scanning along nonstandard spiral and saddle curves, as well as in an n-PI window setting. The algorithmic implementation and numerical results are described to support the correctness of our general claim.

A unified framework for exact cone-beam reconstruction formulas.

In this paper, we present concise proofs of several recently developed exact cone-beam reconstruction methods in the Tuy inversion framework, including both filtered-backprojection and backprojection-filtration formulas in the cases of standard spiral, nonstandard spiral, and more general scanning loci. While a similar proof of the Katsevich formula was previously reported, we present a new proof of the Zou and Pan backprojection-filtration formula. Our proof combines both odd and even data extensions so that only the cone-beam transform itself is utilized in the backprojection-filtration inversion. More importantly, our formulation is valid for general smooth scanning curves, in agreement with an earlier paper from our group [Ye, Zhao, Yu, and Wang, Proc. SPIE 5535, 293-300 (Aug. 6 2004)]. As a consequence of that proof, we obtain a new inversion formula, which is in a two-dimensional filtering backprojection format. A possibility for generalization of the Katsevich filtered-backprojection reconstruction method is also discussed from the viewpoint of this framework.

A backprojection-filtration algorithm for nonstandard spiral cone-beam CT with an n-PI-window.

For applications in bolus-chasing computed tomography (CT) angiography and electron-beam micro-CT, the backprojection-filtration (BPF) formula developed by Zou and Pan was recently generalized by Ye et al to reconstruct images from cone-beam data collected along a rather flexible scanning locus, including a nonstandard spiral. A major implication of the generalized BPF formula is that it can be applied for n-PI-window-based reconstruction in the nonstandard spiral scanning case. In this paper, we design an n-PI-window-based BPF algorithm, and report the numerical simulation results with the 3D Shepp-Logan phantom and Defrise disk phantom. The proposed BPF algorithm consists of three steps: cone-beam data differentiation, weighted backprojection and inverse Hilbert filtration. Our simulated results demonstrate the feasibility and merits of the proposed algorithm.

Design, analysis and simulation for development of the first clinical micro-CT scanner.

In this article, we propose to develop the first clinical micro-CT (CMCT) system for human temporal bone imaging in vivo. This CMCT system consists of medical CT and micro-CT scanners either as separate components or in a combination, a cross-modality registration mechanism such as a facial surface scanner, and associated software. This system integrates the strengths of state-of-the-art medical CT and micro-CT techniques to achieve a spatial resolution that is much higher than currently available for inner ear imaging at acceptable dose levels. Our design, analysis, and simulation results demonstrate that the CMCT system is feasible for inner ear imaging and other clinical applications. For example, the CMCT system has the potential to improve the safety of guiding cochlear implant electrodes within the inner ear and assist the placement of inner ear microcatheters for delivery of gene modification therapy or administration of neurotrophic factors. Imaging of microarchitectures of the cancellous bone would be also an important application.

A knowledge-based cone-beam x-ray CT algorithm for dynamic volumetric cardiac imaging.

With the introduction of spiral/helical multislice CT, medical x-ray CT began a transition into cone-beam geometry. The higher speed, thinner slice, and wider coverage with multislice/cone-beam CT indicate a great potential for dynamic volumetric imaging, with cardiac CT studies being the primary example. Existing ECG-gated cardiac CT algorithms have achieved encouraging results, but they do not utilize any time-varying anatomical information of the heart, and need major improvements to meet critical clinical needs. In this paper, we develop a knowledge-based spiral/helical multislice/cone-beam CT approach for dynamic volumetric cardiac imaging. This approach assumes the relationship between the cardiac status and the ECG signal, such as the volume of the left ventricle as a function of the cardiac phase. Our knowledge-based cardiac CT algorithm is evaluated in numerical simulation and patient studies. In the patient studies, the cardiac status is estimated initially from ECG data and subsequently refined with reconstructed images. Our results demonstrate significant image quality improvements in cardiac CT studies, giving clearly better clarity of the chamber boundaries and vascular structures. In conclusion, this approach seems promising for practical cardiac CT screening and diagnosis.