Design and preliminary evaluation of a multi-channel multi-energy point integrated Wolter microscope.

In inertial confinement fusion, high-precision x-ray imaging is crucial for understanding the states of matter under extreme conditions. To observe the target asymmetry during compression, multiple imaging spots with varying energy responses are necessary. However, integrating multiple imaging channels in a grazing incidence x-ray imaging scheme is challenging, and there is an urgent need for effective combination of multi-color and high-throughput diagnostics. This study presents a design method for a multi-channel integrated Wolter microscope with high spatial resolution, wide-band response, and high throughput. The basic optical configuration, adjustment method, and multi-channel integration scheme are discussed in detail. A 10 keV-class three-channel integrated Wolter microscope is proposed for verification, with an estimated spatial resolution better than 4.0 µm in a field of view of ±500µm. The peak response efficiencies for each imaging channel are calculated as 5.2×10-5 s r, 8.6×10-5 s r, and 2.2×10-4 s r, respectively.

Basic principles and optical system design of 17.48 keV high-throughput modified Wolter x-ray microscope.

High-precision x-ray imaging diagnostics of hotspot at the stagnation stage are essential for regulating implosion asymmetry and retrieving physical implosion parameters. With regard to 10-20 keV energy band imaging, existing diagnostic instruments such as Kirkpatrick-Baez microscopes and pinhole cameras are insufficient in terms of spatial resolution and collection efficiency. The situation is even worse when high-speed, time-resolved imaging diagnostics are performed by coupling framing cameras or line-of-sight imagers. This article presents the basic principles and optical system design of a 17.48 keV modified Wolter x-ray microscope, to resolve the problems encountered in high-energy imaging diagnostics. The proposed optical configuration offers a better spatial resolution, greater depth of field, and preliminary compliance with the requirements of high precision optical processing techniques. The spatial resolution is better than 1 µm in a field range ±150 µm, and is better than 3 µm in a total field of view ∼408 µm in diameter. The geometric solid angle is calculated as 3.0 × 10-5 sr and is estimated to be 1.2 × 10-6 sr, considering the reflectivity of the double mirrors. The proposed microscope is expected to effectively improve spatial resolution and signal-to-noise ratio for high-energy imaging diagnostics.

Design and fabrication of r-hirudin loaded dissolving microneedle patch for minimally invasive and long-term treatment of thromboembolic disease.

Cardiovascular disease is the leading cause of global mortality, with anticoagulant therapy being the main prevention and treatment strategy. Recombinant hirudin (r-hirudin) is a direct thrombin inhibitor that can potentially prevent thrombosis via subcutaneous (SC) and intravenous (IV) administration, but there is a risk of haemorrhage via SC and IV. Thus, microneedle (MN) provides painless and sanitary alternatives to syringes and oral administration. However, the current technological process for the micro mould is complicated and expensive. The micro mould obtained via three-dimensional (3D) printing is expected to save time and cost, as well as provide a diverse range of MNs. Therefore, we explored a method for MNs array model production based on 3D printing and translate it to micro mould that can be used for fabrication of dissolving MNs patch. The results show that r-hirudin-loaded and hyaluronic acid (HA)-based MNs can achieve transdermal drug delivery and exhibit significant potential in the prevention of thromboembolic disease without bleeding in animal models. These results indicate that based on 3D printing technology, MNs combined with r-hirudin are expected to achieve diverse customizable MNs and thus realize personalized transdermal anticoagulant delivery for minimally invasive and long-term treatment of thrombotic disease.

T Lymphocyte Infiltration in Association with IDO1 Expression in Resected Lung Adenocarcinoma and Normal Adjacent Lung Tissues.

BACKGROUND: Indoleamine 2,3-dioxygenase 1 (IDO1) catalyzes the first step of tryptophan catabolism in the kynurenine (Kyn) pathway. IDO1 downregulates natural killer cell receptors, and by mechanism, tumor cells escape immune surveillance. METHODS: IDO1 protein and mRNA were assessed by immunohistochemistry, immunoblotting, and PCR in the 68 resected lung adenocarcinomas at stages I-III as well as adjacent normal lung tissues. Infiltration of CD3, CD8, and CD4 lymphocytes in the tumor and adjacent normal lung tissues was assessed by immunohistochemical staining. RESULTS: IDO1 protein and mRNA were detected in various stages of lung adenocarcinoma with highest expression at stage III. In contrast, biomarkers of T cell subset, CD3, CD4, and CD8, were highly expressed in the normal lung tissues and stage I adenocarcinoma tissues but significantly reduced in the stage II and III tumor tissues. CONCLUSIONS: The current study demonstrated that the higher level of IDO1 expression in the lung adenocarcinoma was, the less infiltration of T lymphocytes was found in the tumors. Findings of this study indicated that IDO1 may contribute to the reduction of T lymphocyte infiltration into the lung adenocarcinoma.

A novel oncotherapy strategy: Direct thrombin inhibitors suppress progression, dissemination and spontaneous metastasis in non-small cell lung cancer.

BACKGROUND AND PURPOSE: Cancer cachexia and cancer-associated thrombosis are potentially fatal outcomes of advanced cancer. Nevertheless, thrombin expression in non-small cell lung cancer (NSCLC) primary tumour tissues and the association between prognosis of NSCLC patients remain largely unknown. EXPERIMENTAL APPROACH: Clinical pathological analysis was performed to determine the relationship between thrombin and tumour progression. Effects of r-hirudin and direct thrombin inhibitor peptide (DTIP) on cancer progression were evaluated. Western blotting, immunohistochemistry, and immunofluorescence were used to explore the inhibition mechanism of r-hirudin and DTIP. The therapeutic effect of the combination of DTIP and chemotherapy was determined. KEY RESULTS: Thrombin expression in NSCLC tissues was closely related to clinicopathological features and the prognosis of patients. Thrombin deficiency inhibited tumour progression. The novel thrombin inhibitors, r-hirudin and DTIP, inhibited cell invasion and metastasis in vitro. They inhibited tumour growth and metastasis in orthotopic lung cancer model, inhibited cell invasion, and prolonged survival after injection of tumour cells via the tail vein. They also inhibited angiogenesis and spontaneous metastases from subcutaneously inoculated tumours. The promotion by thrombin of invasion and metastasis was abolished in PAR-1-deficient NSCLC cells. r-hirudin and DTIP inhibited tumour progression through the thrombin-PAR-1-mediated RhoA and NF-κB signalling cascades via inhibiting MMP9 and IL6 expression. DTIP potentiated chemotherapy-induced growth and metastatic inhibition and inhibited chemotherapy-induced resistance in mice. CONCLUSIONS AND IMPLICATIONS: Thrombin makes a substantial contribution, together with PAR-1, to NSCLC malignancy. The anti-coagulants, r-hirudin and DTIP, could be used in anti-tumour therapy and a combination of DTIP and chemotherapy might improve therapeutic effects.

Orexin dual receptor antagonists, zolpidem, zopiclone, eszopiclone, and cognitive research: A comprehensive dose-response meta-analysis.

Background: About one-third of adults have trouble sleeping, ranging from occasional difficulty to chronic insomnia, along with difficulty maintaining sleep. Many studies reported that the long-term use of hypnotics can cause brain dysfunction and damage cognition. Objective: The objective of the study is to evaluate whether low, medium, and high doses of orexin dual receptor antagonists (DORA), zopiclone (ZOP), eszopiclone (ESZ), and zolpidem (ZST) can impair cognition. Methods: From the beginning through September 20, 2022, PubMed, Embase, Scopus, the Cochrane Library, and Google Scholar were searched. Randomized controlled trials (RCTs) assessing the therapeutic effects of DORA, eszopiclone, and zopiclone for sleep and cognitive function were included. The primary outcomes were indices related to the cognitive profile, including memory, alertness, execution and control function, and attention and orientation. The secondary outcomes were indices related to sleep and adverse events. The standard mean difference (SMD) was generated for continuous variables. Certain data were captured from figures by GetData 2.26 and analyzed using RStudio 4.2. Results: Finally, a total of 8,702 subjects were included in 29 studies. Compared with the placebo, the DSST (Digit Symbol Substitution Test) scores of low, medium, and high doses of DORA were SMD = 0.77; 95% CI: 0.33-1.20; SMD = 1.58; 95% CI: 1.11-2.05; and SMD = 0.85; 95% CI: 0.33-1.36, respectively. The DSST scores of zolpidem at low, medium, and high doses were SMD = -0.39; 95% CI: 0.85-0.07; SMD = -0.88, 95% CI: -2.34-0.58; and SMD = -0.12, 95% CI: -0.85-0.60, respectively. Zopiclone's DSST scale score was SMD = -0.18; 95% CI: -0.54-0.18. In addition, the total sleep time (TST) of low, medium, and high doses of DORA was SMD = 0.28, 95% CI: -0.15-0.70; SMD = 1.36, 95% CI: 0.87-1.86; and SMD = 2.59, 95% CI: 1.89-3.30, respectively. The TST of zolpidem with low, medium, and high doses was SMD = 1.01, 95% CI: 0.18-1.83; SMD = 1.94, 95% CI: 0.46-3.43; and SMD = 1.71, 95% CI: 0.86-2.56, respectively. The TST of low, medium, and high doses of eszopiclone was relatively SMD = 2.03, 95% CI: -0.21-4.27; SMD = 2.38, 95% CI: 1.35-3.42; and SMD = 1.71, 95% CI: 0.60-2.82. Zopiclone's TST was SMD = 2.47, 95% CI: 1.36-3.58. Conclusion: We recommend DORA as the best intervention for insomnia because it is highly effective in inducing and maintaining sleep without impairing cognition. Although zolpidem has a more pronounced effect on maintaining sleep, it is best to reduce its use because of its side effects. Eszopiclone and zopiclone improved sleep quality, but their safety in cognition remains to be verified.

Quantitative observation of monochromatic X-rays emitted from implosion hotspot in high spatial resolution in inertial confinement fusion.

In inertial confinement fusion, quantitative and high-spatial resolution ([Formula: see text]m) measurements of the X-rays self-emitted by the hotspot are critical for studying the physical processes of the implosion stagnation stage. Herein, the 8 ± 0.39-keV monochromatic X-ray distribution from the entire hotspot is quantitatively observed in 5-[Formula: see text]m spatial resolution using a Kirkpatrick-Baez microscope, with impacts from the responses of the diagnosis system removed, for the first time, in implosion experiments at the 100 kJ laser facility in China. Two-dimensional calculations along with 2.5% P2 drive asymmetry and 0.3 ablator self-emission are congruent with the experimental results, especially for the photon number distribution, hotspot profile, and neutron yield. Theoretical calculations enabled a better understanding of the experimental results. Furthermore, the origins of the 17.81% contour profile of the deuterium-deuterium hotspot and the accurate Gaussian source approximation of the core emission area in the implosion capsule are clarified in detail. This work is significant for quantitatively exploring the physical conditions of the hotspot and updating the theoretical model of capsule implosion.

Thrombin is a therapeutic target for non-small-cell lung cancer to inhibit vasculogenic mimicry formation.

Tumor cells transform into endothelial cells by epithelial-to-mesenchymal transition, which is characterized by vasculogenic mimicry (VM). VM not only accelerates tumor progression but also increases drug-induced resistance. However, very little is currently known about the molecular determinants that enable VM. Targeting VM might bring a new breakthrough in cancer treatment. Thrombin is the key enzyme of the blood coagulation system and could contribute to tumor progression. Nevertheless, the association between thrombin and VM formation remains largely unknown. We found that VM was associated with the overall survival of non-small-cell lung cancer (NSCLC) patients, and that thrombin expression was closely related to VM formation. This research revealed that thrombin induced VM formation via PAR-1-mediated NF-κB signaling cascades. The novel thrombin inhibitors r-hirudin and DTIP inhibited VM formation and spontaneous metastases in subcutaneous tumors. Clinical pathological analysis confirmed that NSCLC patients with thrombin-positive/PAR-1-high expression had the poorest prognosis and were the most likely to form VM. The promotional activity of thrombin in VM formation and tumor metastasis was abolished in PAR-1-deficient NSCLC cells. The EGFR inhibitor gefitinib had no effect on VM and increased VEGF expression in tumors. The combination therapy of DTIP and gefitinib achieved a better therapeutic effect than either agent alone. This study is the first to illustrate that thrombin substantially contributes, together with PAR-1, to VM formation and to illustrate that VM might be a target of r-hirudin and DTIP to suppress tumor progression. The anticoagulants r-hirudin and DTIP could be employed for antitumor therapy. Combination therapy with DTIP with an EGFR inhibitor might achieve superior therapeutic effects.

RGD-hirudin-based low molecular weight peptide prevents blood coagulation via subcutaneous injection.

Thromboembolic disease is a common cardio-cerebral vascular disease that threatens human life and health. Thrombin not only affects the exogenous coagulation pathway, but also the endogenous pathway. Thus, it becomes one of the most important targets of anticoagulant drugs. RGD-hirudin is an anticoagulant drug targeting thrombin, but it can only be administered intravenously. We designed a low molecular weight peptide based on RGD-hirudin that could prevent blood clots. We first used NMR to identify the key amino acid residues of RGD-hirudin that interacted with thrombin. Then, we designed a novel direct thrombin inhibitor peptide (DTIP) based on the structure and function of RGD-hirudin using homology modeling. Molecular docking showed that the targeting and binding of DTIP with thrombin were similar to those of RGD-hirudin, suggesting DTIP interacted directly with thrombin. The active amino acids of DTIP were identified by alanine scanning, and mutants were successfully constructed. In blood clotting time tests in vitro, we found that aPTT, PT, and TT in the rat plasma added with DTIP were greatly prolonged than in that added with the mutants. Subcutaneous injection of DTIP in rats also could significantly prolong the clotting time. Thrombelastography analysis revealed that DTIP significantly delayed blood coagulation. Bio-layer interferometry study showed that there were no significant differences between DTIP and the mutants in thrombin affinity constants, suggesting that it might bind to other sites of thrombin rather than to its active center. Our results demonstrate that DTIP with low molecular weight can prevent thrombosis via subcutaneous injection.

RhTyrRS (Y341A), a novel human tyrosyl-tRNA synthetase mutant, stimulates thrombopoiesis through activation of the VEGF-R II/NF-κB pathway.

BACGROUND AND PURPOSE: Tumor chemotherapy and radiotherapy induces hematopoietic cell damage, resulting in thrombocytopenia. Conventional platelet transfusion strategies or drug therapies are used to treat thrombocytopenia. However, these therapies may result in a several side effects, including heightened susceptibility to infectious diseases and the formation of anti-TPO-antibodies. Therefore, a more secure strategy should be explored to overcome and compensate for the shortcomings of conventional strategies. EXPERIMENTAL APPROACH: Effects of rhTyrRS(Y341A) on the expression of VCAM-1 on the surface of HUVECs were determined by analysing mRNA expression, promoter activity, protein expression. The molecular mechanisms of the effects of rhTyrRS(Y341A) on the expression of VCAM-1 on the surface of HUVECs were investigated by determining the activation of VEGF-R II/NF-κB pathway. KEY RESULTS: Our results provide evidence that rhTyrRS (Y341A) activates NF-κB to upregulate VCAM-1 in a VEGF-R II/NF-κB pathway-dependent, resulting in megakaryocyte adhering to PVECs to induce platelet production. CONCLUSIONS: This study suggested that rhTyrRS (Y341A), a novel human tyrosyl-tRNA synthetase mutation, increased the platelet count under normal conditions. Further more, we confirmed that an NF-κB-mediated mechanism is involved in rhTyrRS (Y341A)-induced thrombopoiesis, which involves its interaction with VEGF-R II.

Development of a polar-view Kirkpatrick-Baez X-ray microscope for implosion asymmetry studies.

The development of a polar-view Kirkpatrick-Baez microscope, fielded in the upper polar zone of the Shenguang-III laser fusion facility, is presented. With this microscope, the resolving power of polar-direction X-ray imaging diagnostics is improved, to the 3 ~5 µm scale. The microscope is designed for implosion asymmetry studies, with response energy points at 1.2 keV, 3.5 keV, and 8 keV. A biperiodic multilayer scheme is adopted to accommodate multiple implosion stages. We present the overall optical system design, target aiming scheme, characteristic composite imaging diagnostic experiments and initial results. The inertial-driven quasi-one-dimensional spherical implosions were observed from orthogonal directions with a convergence ratio of ~14.4. Fine features of the stagnating hot spot core are also resolved.

Note: New method for high-space-resolving hotspot electron temperature measurements on Shenguang-III prototype.

High-space-resolving information of hotspot electron temperature is a foundation for further research on physical processes of implosion in inertial confinement fusion. This work proposed a novel high-space-resolving electron temperature detector, which is based on the bremsstrahlung radiation mechanism of the implosion hotspot and uses two-channel Kirkpatrick-Baez microscopes. In this novel detector, an optical quasi-coaxis method was used to eliminate the strong impact of the view field difference on the high space resolution and correctness of the electron temperature diagnosis, and a compound KB microscope method was proposed to reduce the number of spherical reflectors and save space.

Direct intensity calibration of X-ray grazing-incidence microscopes with home-lab source.

Direct intensity calibration of X-ray grazing-incidence microscopes is urgently needed in quantitative studies of X-ray emission from laser plasma sources in inertial confinement fusion. The existing calibration methods for single reflecting mirrors, crystals, gratings, filters, and X-ray detectors are not applicable for such X-ray microscopes due to the specific optical structure and the restrictions of object-image relation. This article presents a reliable and efficient method that can be performed using a divergent X-ray source and an energy dispersive Si-PIN (silicon positive-intrinsic-negative) detector in an ordinary X-ray laboratory. The transmission theory of X-ray flux in imaging diagnostics is introduced, and the quantities to be measured are defined. The calibration method is verified by a W/Si multilayer-coated Kirkpatrick-Baez microscope with a field of view of ∼95 µm at 17.48 keV. The mirror reflectance curve in the 1D coordinate is drawn with a peak value of 20.9% and an uncertainty of ∼6.0%.

Development of an x-ray eight-image Kirkpatrick-Baez diagnostic system for China's laser fusion facility.

This article presents the development of an x-ray eight-image Kirkpatrick-Baez diagnostic system to be used at China's Shenguang-III (SG-III) laser facility in aspects of the optical design, multilayers, and online/offline tests. Six pieces of concave spherical substrates are used for constituting a special optical structure. Dual-periodic tungsten/carbon (W/C) multilayers are used for high reflectivity and large angular bandwidth of ∼0.1°. The global spatial resolution is ∼5 µm in the ±100 µm range. The schemes of system installation, transport, collimation, and image acquisition at China's SG-III facility are also discussed.

A novel hirudin derivative inhibiting thrombin without bleeding for subcutaneous injection.

Currently, anticoagulants would be used to prevent thrombosis. Thrombin is an effector enzyme for haemostasis and thrombosis. We designed a direct thrombin inhibitor peptide (DTIP) using molecular simulation and homology modelling and demonstrated that the C-terminus of DTIP interacts with exosite I, and N-terminus with the activity site of thrombin, respectively. DTIP interfered with thrombin-mediated coagulation in human, rat and mouse plasma (n=10 per group) and blocked clotting in human whole blood in vitro. When administered subcutaneously, DTIP showed potent and dose-dependent extension of aPTT, PT, TT and CT in rats (n=10 per group). The antithrombotic dose of DTIP induced significantly less bleeding than bivalirudin determined by transecting distal tail assay in rats. Furthermore, DTIP reached peak blood concentration in 0.5-1 hour and did not cause increased bleeding after five days of dosing compared to dabigatran etexilate. The antithrombotic effect of DTIP was evaluated in mice using lethal pulmonary thromboembolism model and FeCl3-induced mesenteric arteriole thrombus model. DTIP (1.0 mg/kg, sc) prevented deep venous thrombosis and increased the survival rate associated with pulmonary thromboembolism from 30 % to 80 %. Intravital microscopy showed that DTIP (1.0 mg/kg, sc) decelerated mesenteric arteriole thrombosis caused by FeCl3 injury. These data establish that DTIP is a novel antithrombotic agent that could be used to prevent thrombosis without conferring an increased bleeding risk.

Development of high resolution dual-energy KBA microscope with large field of view for RT-instability diagnostics at SG-III facility.

High resolution X-ray diagnosis is a significant method for obtaining ablation-front and trajectory measurements targeting Rayleigh-Taylor (RT)-instability growth in initial confinement fusion (ICF) experiments. In this paper, a novel Kirkpatrick-Baez-type structure, as a kind of essential X-ray micro-imaging apparatus, has been developed that realizes a large field of view (FOV) and images with high resolution and energy response. Zoned multilayer coating technology is applied to the Kirkpatrick-Baez mirrors to transmit two specific quasi-monochromatic light through the same mirror and enables a compact dual-channel structure. This microscope has been assembled in the laboratory and later implemented at the Chinese SG-III laser facility. The characterization results show that this imaging system can achieve a good spatial resolution of 5 µm in a large FOV of 500 µm, while maintaining a strong monochromatic performance with bandwidth of 0.5 keV at 2.5 keV and 4.3 keV respectively.