A novel dilated contextual attention module for breast cancer mitosis cell detection.

Background and object: Mitotic count (MC) is a critical histological parameter for accurately assessing the degree of invasiveness in breast cancer, holding significant clinical value for cancer treatment and prognosis. However, accurately identifying mitotic cells poses a challenge due to their morphological and size diversity. Objective: We propose a novel end-to-end deep-learning method for identifying mitotic cells in breast cancer pathological images, with the aim of enhancing the performance of recognizing mitotic cells. Methods: We introduced the Dilated Cascading Network (DilCasNet) composed of detection and classification stages. To enhance the model's ability to capture distant feature dependencies in mitotic cells, we devised a novel Dilated Contextual Attention Module (DiCoA) that utilizes sparse global attention during the detection. For reclassifying mitotic cell areas localized in the detection stage, we integrate the EfficientNet-B7 and VGG16 pre-trained models (InPreMo) in the classification step. Results: Based on the canine mammary carcinoma (CMC) mitosis dataset, DilCasNet demonstrates superior overall performance compared to the benchmark model. The specific metrics of the model's performance are as follows: F1 score of 82.9%, Precision of 82.6%, and Recall of 83.2%. With the incorporation of the DiCoA attention module, the model exhibited an improvement of over 3.5% in the F1 during the detection stage. Conclusion: The DilCasNet achieved a favorable detection performance of mitotic cells in breast cancer and provides a solution for detecting mitotic cells in pathological images of other cancers.

Immunologic barriers in liver transplantation: a single-cell analysis of the role of mesenchymal stem cells.

Background: This study aimed to analyze the biomarkers that may reliably indicate rejection or tolerance and the mechanism that underlie the induction and maintenance of liver transplantation (LT) tolerance related to immunosuppressant or mesenchymal stem cells (MSCs). Methods: LT models of Lewis-Lewis and F344-Lewis rats were established. Lewis-Lewis rats model served as a control (Syn). F344-Lewis rats were treated with immunosuppressant alone (Allo+IS) or in combination with MSCs (Allo+IS+MSCs). Intrahepatic cell composition particularly immune cells was compared between the groups by single-cell sequencing. Analysis of subclusters, KEGG pathway analysis, and pseudotime trajectory analysis were performed to explore the potential immunoregulatory mechanisms of immunosuppressant alone or combined with MSCs. Results: Immunosuppressants alone or combined with MSCs increases the liver tolerance, to a certain extent. Single-cell sequencing identified intrahepatic cell composition signature, including cell subpopulations of B cells, cholangiocytes, endothelial cells, erythrocytes, hepatic stellate cells, hepatocytes, mononuclear phagocytes, neutrophils, T cells, and plasmacytoid dendritic cells. Immunosuppressant particularly its combination with MSCs altered the landscape of intrahepatic cells in transplanted livers, as well as gene expression patterns in immune cells. MSCs may be included in the differentiation of T cells, classical monocytes, and non-classical monocytes. Conclusion: These findings provided novel insights for better understanding the heterogeneity and biological functions of intrahepatic immune cells after LT treated by IS alone or in combination with MSCs. The identified markers of immune cells may serve as the immunotherapeutic targets for MSC treatment of liver transplant rejection.

Aerosol Inhalation of Chimpanzee Adenovirus Vectors (ChAd68) Expressing Ancestral or Omicron BA.1 Stabilized Pre-Fusion Spike Glycoproteins Protects Non-Human Primates against SARS-CoV-2 Infection.

Current COVID-19 vaccines are effective countermeasures to control the SARS-CoV-2 virus pandemic by inducing systemic immune responses through intramuscular injection. However, respiratory mucosal immunization will be needed to elicit local sterilizing immunity to prevent virus replication in the nasopharynx, shedding, and transmission. In this study, we first compared the immunoprotective ability of a chimpanzee replication-deficient adenovirus-vectored COVID-19 vaccine expressing a stabilized pre-fusion spike glycoprotein from the ancestral SARS-CoV-2 strain Wuhan-Hu-1 (BV-AdCoV-1) administered through either aerosol inhalation, intranasal spray, or intramuscular injection in cynomolgus monkeys and rhesus macaques. Compared with intranasal administration, aerosol inhalation of BV-AdCoV-1 elicited stronger humoral and mucosal immunity that conferred excellent protection against SARS-CoV-2 infection in rhesus macaques. Importantly, aerosol inhalation induced immunity comparable to that obtained by intramuscular injection, although at a significantly lower dose. Furthermore, to address the problem of immune escape variants, we evaluated the merits of heterologous boosting with an adenovirus-based Omicron BA.1 vaccine (C68-COA04). Boosting rhesus macaques vaccinated with two doses of BV-AdCoV-1 with either the homologous or the heterologous C68-COA04 vector resulted in cross-neutralizing immunity against WT, Delta, and Omicron subvariants, including BA.4/5 stronger than that obtained by administering a bivalent BV-AdCoV-1/C68-COA04 vaccine. These results demonstrate that the administration of BV-AdCoV-1 or C68-COA04 via aerosol inhalation is a promising approach to prevent SARS-CoV-2 infection and transmission and curtail the pandemic spread.

Cu(I) complexes with aggregation-induced emission for enhanced photodynamic antibacterial application.

This communication reports the design of aggregation-induced emission (AIE)-featured PEG-condensed Cu(I)-p-MBA aggregates (PCuA). Benefiting from the AIE trait and intrinsic antibacterial property of Cu species, the as-developed PCuA exhibits enhanced photodynamic antibacterial activities against broad-spectrum bacteria, providing a paradigm in the design of novel antibacterial agents.

Achieving high intracellular trehalose in hRBCs by reversible membrane perturbation of maltopyranosides with synergistic membrane protection of macromolecular protectants.

Trehalose is considered as a biocompatible cryoprotectant for solvent-free cryopreservation of cells, but the difficulty of the current trehalose delivery platforms to human red blood cells (hRBCs) limits its wide applications. Due to cell injuries caused by incubation at 37 °C and low intracellular loading efficiency, development of novel methods to facilitate trehalose entry in hRBCs is essential. Herein, a reversible membrane perturbation and synergistic membrane stabilization system based on maltopyranosides and macromolecular protectants was constructed, demonstrating the ability of efficient trehalose loading in hRBCs at 4 °C. Results of confocal laser scanning microscopy exhibited that the intracellular loading with the assistance of maltopyranosides was a reversible process, while the membrane protective effect of macromolecular protectants on trehalose loading in hRBCs was necessary. It was suggested that introduction of 30 mM poly(vinyl pyrrolidone) 8000 combined with 1 mM dodecyl-ß-D-maltopyranoside and 0.8 M trehalose could increase the intracellular trehalose to 84.0 ± 11.3 mM in hRBCs, whereas poly(ethylene glycol), dextran, human serum albumin or hydroxyethyl starch had a weak effect. All the macromolecular protectants could promote the cryosurvival of hRBCs, exhibiting membrane stabilization, and incubation and followed by cryopreservation did not change the basic functions and normal morphology of hRBCs substantially. This study provided an alternative strategy for glycerol-free cryopreservation of cells and the delivery of membrane-impermeable cargos.

Membrane stabilization versus perturbation by aromatic monoamine-modified γ-PGA for cryopreservation of human RBCs with high intracellular trehalose.

As a nonreducing disaccharide, trehalose can be used as a biocompatible cryoprotectant for solvent-free cell cryopreservation, but the membrane-impermeability limits its cryoprotective efficiency. Herein, a series of aromatic monoamines with a 1-4 methylene spacer were grafted onto γ-poly(glutamic acid) (γ-PGA) for promoting intracellular trehalose uptake in human red blood cells (hRBCs) via membrane perturbation. The self-assembled nanoparticles of the obtained amphiphilic γ-PGA could be adsorbed on the cell membrane by the hydrophobic interaction to disturb the lipid arrangement and increase the membrane permeability of trehalose under hypertonic conditions. Results suggested that the intracellular trehalose could be enhanced progressively with the methylene spacer length, significantly increasing to 75.1 ± 0.7 mM by incubating hRBCs in 0.8 M trehalose containing phenylbutylamine-grafted γ-PGA at 4 °C for 24 h. Meanwhile, the other three polymers exhibited membrane stabilization in addition to improved intracellular trehalose, maintaining the membrane integrity during cryopreservation to achieve high cryosurvival. Molecular dynamics simulation further confirmed that defects could be formed by interaction of the above four amphiphilic polymers on the modeled phospholipid bilayer. It was believed that glycerol-free cryopreservation of human cells could be realized by using trehalose as the biocompatible cryoprotectant, and membrane stabilization can be a compensatory approach to membrane perturbation during impermeable biomolecule delivery.

Cryopreservation of human erythrocytes through high intracellular trehalose with membrane stabilization of maltotriose-grafted ε-poly(L-lysine).

Cryopreservation of human erythrocytes via suitable cryoprotectants is essential for transfusion during emergencies, but the conventional glycerolization method requires a tedious thawing-deglycerolization process. Alternatively, trehalose, a nonreducing disaccharide, has gained much attention as a biocompatible cryoprotectant due to its nature in living organisms capable of surviving extreme cold and desiccation. In this work, cryopreservation of human erythrocytes was realized through high intracellular trehalose enhanced by benzyl alcohol at 4 °C with membrane stabilization of maltotriose-grafted ε-poly(L-lysine). Intracellular trehalose could reach 94.2 ± 12.1 mM with slight impacts on morphology and cell functions, and the post-storage cryosurvival of human erythrocytes could achieve 96.2 ± 3.4% via membrane protection by the glycopeptide. It has been demonstrated that the functional glycopeptide performed as an extracellular cryoprotectant accompanied by high intracellular trehalose for synergistic cryopreservation of human erythrocytes in the biocompatible glycerol-free conditions. This two-step approach involving augmentation of intracellular trehalose at a hypothermic temperature and membrane stabilization of the functional glycopeptide could be an alternative way for human cell cryopreservation.

Facilitating trehalose entry into hRBCs at 4 °C by alkylated ε-poly(L-lysine) for glycerol-free cryopreservation.

Currently, glycerol is a conventional cryoprotectant of human red blood cells (hRBCs), but the time-consuming thawing and deglycerolization processes are essential before transfusion. Much of the research up to now has been conducted on the delivery of impermeable trehalose to hRBCs at 37 °C, but the cryoprotective effect of trehalose and deterioration of cells still remain challenging. Encouraged by the interaction of hydrophobic or cationic groups on cell membranes and osmotic stabilization, herein, we propose a novel cryopreservation system to facilitate trehalose entry into hRBCs at 4 °C and pH 7.4. High intracellular trehalose contents and cryosurvival of hRBCs were achieved with small function variations via the assistance of self-assembled nanoparticles of alkylated ε-poly(L-lysine) (ε-PL) along with poly(vinyl pyrrolidone) (PVP). The effect of amphipathic alkylated ε-PL with various alkyl chains and grafting ratios on membrane perturbation with protection of PVP was systematically investigated. Overall, by the combination of alkylated ε-PL and PVP, the intracellular trehalose could be enhanced to 109.7 ± 6.1 mM and subsequently hRBC cryosurvival reached 91.7 ± 5.5%, significantly higher than those containing trehalose only, 11.9 ± 1.1 mM and 50.0 ± 2.1%, respectively. It was observed that the biocompatible trehalose-loading system could benefit glycerol-free cryopreservation of hRBCs and also provide a feasible way for impermeable biomacromolecule delivery.

Transcriptomic and metabolomic analyses reveal the altitude adaptability and evolution of different-colored flowers in alpine Rhododendron species.

Understanding the molecular mechanisms and evolutionary process of plant adaptation to the heterogeneous environment caused by altitude gradients in plateau mountain ecosystems can provide novel insight into species' responses to global changes. Flower color is the most conspicuous and highly diverse trait in nature. Herein, the gene expression patterns, evolutionary adaptation and metabolites changes of different-colored flowers of alpine Rhododendron L. species along altitude gradients were investigated based on a combined analysis of transcriptomics and metabolomics. Differentially expressed genes were found to be related to the biosynthesis of carbohydrates, fatty acids, amino acids and flavonoids, suggesting their important roles in the altitude adaptability of Rhododendron species. The evolution rate of high-altitude species was faster than that of low-altitude species. Genes related to DNA repair, mitogen-activated protein kinase and ABA signal transduction, and lipoic acid and propanoate metabolism were positively selected in the flowers of high-altitude Rhododendron species and those associated with carotenoid biosynthesis pathway, ABA signal transduction and ethylene signal transduction were positively selected in low-altitude species. These results indicated that the genes with differentiated expressions or functions exhibit varying evolution during the adaptive divergence of heterogeneous environment caused by altitude gradients. Flower-color variation might be attributed to the significant differences in gene expression or metabolites related to sucrose, flavonoids and carotenoids at the transcription or metabolism levels of Rhododendron species. This work suggests that Rhododendron species have multiple molecular mechanisms in their adaptation to changing environments caused by altitude gradients.

Analgesic effect of preoperative dezocine-based local anesthesia in patients undergoing inguinal hernia repair.

OBJECTIVE: This study was performed to investigate the analgesic effects of intramuscular injection of dezocine-based local anesthesia in patients undergoing inguinal hernia repair. METHODS: A total of 120 patients underwent local herniorrhaphy from July 2015 to December 2016 and were randomly divided into 2 groups: the dezocine group, in which a preoperative intramuscular injection of dezocine was administered, and the control group, in which no dezocine injection was administered. The patients' pain and comfort levels were evaluated at 30 minutes, 2 hours, 6 hours, 12 hours, and 24 hours postoperatively. RESULTS: The visual analog scale scores at 30 minutes, 2 hours, 6 hours, 12 hours, and 24 hours were significantly lower in the dezocine than control group. In the Bruggemann comfort scale evaluation, patients who received dezocine injections showed significantly greater comfort than those in the control group at 30 minutes, 2 hours, 6 hours, 12 hours, and 24 hours. No adverse reactions occurred in the dezocine group. CONCLUSIONS: Dezocine-based local anesthesia can attenuate postoperative pain and increase the comfort level during and after herniorrhaphy.

The Transformation of Hybrid Silica Nanoparticles from Solid to Hollow or Yolk-Shell Nanostructures.

Here, a facile self-templating approach is presented for synthesis of hollow and yolk-shell mesoporous silica nanoparticles (HMSNs and YMSNs) through a selective etching of hybrid silica nanoparticles. The hybrid silica nanoparticles are from the co-condensation of tetraethylorthosilicate (TEOS) and N-[3-(trimethoxysilyl)propyl]ethylenediamine (TSD) by a simple one-step process. Two kinds of products including HMSNs and YMSNs can be easily prepared only by tuning the TSD amounts in the precursor. Significantly, the transformation of hollow structure does not use any sacrificial template and surface-protective agent. The etching mechanism and formation process are systematically investigated by SEM, TEM, TG, CHN elemental analysis and Si MAS NMR spectroscopy. The results reveal that the selective etching is mainly attributed to the discrepancy in density between the outer layer and inner area of hybrid silica, where its inner section is more readily dissolved while the outer shell is robust in hydrofluoric acid (HF) aqueous solution. Specifically, the new understanding is further extended to precisely prepare multi-shelled hollow/yolk-shell silica nanoparticles.

Surfactant-assisted selective etching strategy for generation of rattle-like mesoporous silica nanoparticles.

Nowadays, rattle-like or so-called yolk-shell nanostructures have set off a new wave of research in view of their prominent features including large surface area, tunable void and flexible functional core, etc. Herein, rattle-like mesoporous silica nanoparticles (RMSNs) with a pure silica core, a hollow cavity and a mesoporous shell have been successfully fabricated via a surfactant-assisted selective etching strategy. The synthetic approach involves the preparation of solid silica spheres with three-layer different structural silica containing the inner core of pure silica, middle layer of hybrid silica and outer shell of surfactant/SiO2 composite, followed by a hydrothermal treatment in hot water. The resulting products show a distinct rattle-like structure and spherical morphology. The average diameter, the shell thickness, and the solid core size of RMSNs are about 290, 35 and 90nm, respectively. During the etching process, the surfactant with different length of alkyl chain (CnTAB, e.g. cetyltrimethylammonium bromide) in outer shell plays a decisive role for the formation of rattle-like structure. Benefiting from the residual amino groups in RMSNs, Au@RMSNs composites can be further constructed by in-situ generating Au nanoparticles into their hollow cavity, demonstrating an excellent catalytic performance for reduction of 4-nitrophenol. Additionally, RMSNs also show a strong ability for adsorption of rhodamine B.

Stability criteria for T-S fuzzy systems with interval time-varying delays and nonlinear perturbations based on geometric progression delay partitioning method.

In this paper, a novel delay partitioning method is proposed by introducing the theory of geometric progression for the stability analysis of T-S fuzzy systems with interval time-varying delays and nonlinear perturbations. Based on the common ratio α, the delay interval is unequally separated into multiple subintervals. A newly modified Lyapunov-Krasovskii functional (LKF) is established which includes triple-integral terms and augmented factors with respect to the length of every related proportional subintervals. In addition, a recently developed free-matrix-based integral inequality is employed to avoid the overabundance of the enlargement when dealing with the derivative of the LKF. This innovative development can dramatically enhance the efficiency of obtaining the maximum upper bound of the time delay. Finally, much less conservative stability criteria are presented. Numerical examples are conducted to demonstrate the significant improvements of this proposed approach.

Perturbed dynamics of discrete-time switched nonlinear systems with delays and uncertainties.

This paper addresses the dynamics of a class of discrete-time switched nonlinear systems with time-varying delays and uncertainties and subject to perturbations. It is assumed that the nominal switched nonlinear system is robustly uniformly exponentially stable. It is revealed that there exists a maximal Lipschitz constant, if perturbation satisfies a Lipschitz condition with any Lipschitz constant less than the maximum, then the perturbed system can preserve the stability property of the nominal system. In situations where the perturbations are known, it is proved that there exists an upper bound of coefficient such that the perturbed system remains exponentially stable provided that the perturbation is scaled by any coefficient bounded by the upper bound. A numerical example is provided to illustrate the proposed theoretical results.

A facile route for rapid synthesis of hollow mesoporous silica nanoparticles as pH-responsive delivery carrier.

In this paper, a facile and effective route has been developed for rapid synthesis of hollow mesoporous silica nanoparticles (HMSNs) by using tetradecyltrimethylammonium bromide (TTAB) as the porogen with the assistance of triethanolamine (TEA). The products were characterized by various techniques including TEM, SEM, BET, and FT-IR, etc. The HMSNs obtained possess spherical morphology, mesoporous channels and very high specific surface areas (1355m(2)g(-1)). According to the experimental results, a possible formation mechanism was discussed. Moreover, the ability of HMSNs as drug carrier was evaluated by selecting doxorubicin hydrochloride (DOX) as the model drug. The results indicated that HMSNs showed high loading capacity and controlled pH-responsive release behavior. Considering their unique nanostructures and porous properties, we expect the HMSNs prepared have more potential applications in various fields such as nanoreactors, cellular imaging, and biosensor.

Exploration of common bile duct for treating hepatic cystic echinococcosis.

OBJECTIVE: To compare the clinical efficacy of exploration of common bile duct through cystic duct or T type tube in CE patients. METHODS: One hundred and fifty six patients with chronic CE received decompression of biliary tract from January 2007 to December 2012 were included. Exploration of common bile duct was performed through cystic duct (n=102) or T type tube (n=54). Psychological reactions were monitored including inconvenient position alteration due to carrying of the drainage tube, anxiety of the folding and prolapse of drainage tube, folding of the drainage tube, and any pain during the body position change. RESULTS: No significant difference was noticed in the body position alteration discomfort in both groups (P>0.05). Statistical difference was noted in the anxiety of folding and prolapse of draining tube (P<0.05), and improvement of the clinical symptoms (P<0.05). CONCLUSIONS: A higher possibility of wound, enlarged tube-carrying duration, and lower sense of discomfort, were noticed in patients underwent exploration of common bile duct through T type tube. Higher sense of comfort was reported in patients received exploration of common bile duct through cystic duct tube.

[Secondary laryngeal tuberculosis at high altitudes of Tibet].

OBJECTIVE: To improve the diagnostic and therapeutic efficiency for secondary laryngeal tuberculosis through an analysis on the clinical features of patients with this disease. METHOD: A retrospective study was made among 49 cases with laryngeal tuberculosis treated in Tibetan General Hospital of Chinese PLA, and the clinical data were carefully analyzed to summarize the clinical experience of this disease. RESULT: Of 49 patients, 24 cases had 1 year history, 11 cases had 1 to 3 years, 9 cases had 3 to 5 years, 5 cases had 5 years or more. Thirty-eight patients had the history of tuberculosis and 11 had none. Thirty-four patients had taken anti-tuberculosis drugs but none had standard therapy as demanded. All cases had mild general symptoms (mild fever, night sweats, weight loss, et al) and atypical local symptoms (hoarseness, sore throat). Therefore, 42 cases were misdiagnosed as non-specific chronic laryngitis, of which 15 cases got worse after oral administration or inhaling of steroid hormones. Seven persons were misdiagnosed as laryngeal cancer. All patients were confirmed pulmonary tuberculosis by X ray exam or CT scanning. Twelve cases had strong positive PPD tests and 2 cases were detected positive by sputum smear. All patients was treated by standard systematic and local chemical therapy against tuberculosis (inhaling of antituberculosis drugs for 1 to 2 months). All were cured but one died in a road accident, and none had recurrence after 1- to 9- year follow-up. CONCLUSION: All of those the patients with long period hoarseness and sore throat should take chest CT scan or X-ray exam for the highest incidence of pulmonary tuberculosis at high altitudes. CT scanning is the prefer for its high resolution. Pathological biopsy and diagnostic therapy should be taken to make accurate diagnosis. Usually steroid hormones should not be recommended.

[Characteristics of the PAH gene mutation in Chinese patients with phenylketonuria in Xinjiang].

OBJECTIVE: To study the characteristics of the PAH gene mutation in patients with phenylketonuria (PKU) in Xinjiang area. METHODS: The mutations in exons 3, 5, 6, 7, 11 and 12 and the flanking intronic sequence of the PAH gene were detected by PCR/SSCP analysis and direct DNA sequencing in 46 PKU patients. RESULTS: Twenty different mutations were found in 68/92 alleles (73.9%). The prevalent mutations of R243Q, EX6 96A>G, R111X, Y356X and V399V were similar to that of Northern China populations. The mutations F161S, L255S, P281L, and R413P were significantly different from that in other Chinese populations. It was the second time that E280G and A434D mutations were reported in the world, that L255S, P281L, R261Q, and I65T mutations were found in China. Thirteen different mutations were first found in Chinese Uygur, which showed a distinct ethnic characteristics. CONCLUSION: The study showed not only a distinct and conservative, but also a crossed and syncretic genetic characteristics in Xinjiang Uygur population. The results suggest that Xinjiang could be an ideal genetic resource repertoire for studying diversity of gene mutations, heterogeneity of PAH gene, human origins and migration.