A chimeric porcine reproductive and respiratory syndrome virus 1 strain containing synthetic ORF2-6 genes can trigger T follicular helper cell and heterologous neutralizing antibody responses and confer enhanced cross-protection.

The prevalence of porcine reproductive and respiratory syndrome virus 1 (PRRSV1) isolates has continued to increase in Chinese swine herds in recent years. However, no effective control strategy is available for PRRSV1 infection in China. In this study, we generated the first infectious cDNA clone (rHLJB1) of a Chinese PRRSV1 isolate and subsequently used it as a backbone to construct an ORF2-6 chimeric virus (ORF2-6-CON). This virus contained a synthesized consensus sequence of the PRRSV1 ORF2-6 gene encoding all the envelope proteins. The ORF2-6 consensus sequence shared > 90% nucleotide similarity with four representative strains (Amervac, BJEU06-1, HKEU16 and NMEU09-1) of PRRSV1 in China. ORF2-6-CON had replication efficacy similar to that of the backbone rHLJB1 virus in primary alveolar macrophages (PAMs) and exhibited cell tropism in Marc-145 cells. Piglet inoculation and challenge studies indicated that ORF2-6-CON is not pathogenic to piglets and can induce enhanced cross-protection against a heterologous SD1291 isolate. Notably, ORF2-6-CON inoculation induced higher levels of heterologous neutralizing antibodies (nAbs) against SD1291 than rHLJB1 inoculation, which was concurrent with a higher percentage of T follicular helper (Tfh) cells in tracheobronchial lymph nodes (TBLNs), providing the first clue that porcine Tfh cells are correlated with heterologous PRRSV nAb responses. The number of SD1291-strain-specific IFNγ-secreting cells was similar in ORF2-6-CON-inoculated and rHLJB1-inoculated pigs. Overall, our findings support that the Marc-145-adapted ORF2-6-CON can trigger Tfh cell and heterologous nAb responses to confer improved cross-protection and may serve as a candidate strain for the development of a cross-protective PRRSV1 vaccine.

Xylem plasticity of root, stem, and branch in Cunninghamia lanceolata under drought stress: implications for whole-plant hydraulic integrity.

Introduction: A better understanding of xylem hydraulic characteristics in trees is critical to elucidate the mechanisms of forest decline and tree mortality from water deficit. As well as temperate forests and forests growing in arid regions, subtropical and tropical forests are also predicted to experience an increased frequency and intensity of climate change-induced drought in the near future. Methods: In this study, 1-year-old Cunninghamia lanceolata seedlings (a typical subtropical species in southern China) were selected for a continuous controlled drought pot experiment of 45 days duration. The experimental treatments were non-drought (control), light drought, moderate drought and severe drought stress, which were 80%, 60%, 50%, and 40%, respectively of soil field maximum moisture capacity. Results: The hydraulic conductivity, specific conductivity and water potential of roots, stems, and branches of C. lanceolata all decreased with the prolonging of drought in the different drought intensities. The relative decrease in these hydraulic values were greater in roots than in stems and branches, indicating that roots are more sensitive to drought. Root tracheid diameters normally reduce to ensure security of water transport with prolonged drought, whilst the tracheid diameters of stems and branches expand initially to ensure water transport and then decrease to reduce the risk of embolism with continuing drought duration. The pit membrane diameter of roots, stems and branches generally increased to different extents during the 15-45 days drought duration, which is conducive to enhanced radial water transport ability. The tracheid density and pit density of stems generally decreased during drought stress, which decreased water transport efficiency and increased embolism occurrence. Correlation analysis indicated that anatomical plasticity greatly influenced the hydraulic properties, whilst the relationships varied among different organs. In roots, tracheid diameter decreased and tracheid density increased to enhance water transport security; stems and branches may increase tracheid diameter and pit membrane diameter to increase hydraulic conductivity ability, but may increase the occurrence of xylem embolism. Discussion: In summary, under drought stress, the xylem anatomical characteristics of C. lanceolata organs were highly plastic to regulate water transport vertically and radially to maintain the trade-off between hydraulic conductivity efficiency and safety.

Efficacy of Electronic Cigarettes vs Varenicline and Nicotine Chewing Gum as an Aid to Stop Smoking: A Randomized Clinical Trial.

Importance: Electronic cigarettes (ECs) are often used by smokers as an aid to stopping smoking, but evidence is limited regarding their efficacy compared with nicotine replacement therapy (NRT), and no evidence is available on how their efficacy compares with that of varenicline. Objective: To evaluate whether ECs are superior to NRT and noninferior to varenicline in helping smokers quit. Design, Setting, and Participants: This was a randomized clinical trial conducted at 7 sites in China and including participants who were smoking at least 10 cigarettes per day and motivated to quit, not using stop-smoking medications or EC, and willing to use any of the study products. Participants were first recruited in May 2021, and data analysis was conducted in December 2022. Interventions: A cartridge-based EC (30 mg/mL nicotine salt for 2 weeks and 50 mg/mL after that), varenicline (0.5 mg, once a day for 3 days; 0.5 mg, twice a day for 4 days; and 1 mg, twice a day, after that), and 2 mg (for smokers of ≤20 cigarettes per day) or 4 mg (>20 cigarettes per day) nicotine chewing gum, all provided for 12 weeks and accompanied by minimal behavioral support (an invitation to join a self-help internet forum). Main Outcomes and Measures: The primary outcome was sustained abstinence from smoking at 6 months as validated by an expired-air carbon monoxide reading (<8 parts per million). Participants lost to follow-up were included as nonabstainers. Results: Of 1068 participants, 357 (33.5%) were female, and the mean (SD) age was 33.9 (3.1) years. A total of 409 (38.3%), 409 (38.3%), and 250 (23.4%) participants were randomized to the EC, varenicline, and NRT arms, respectively. The 6-month biochemically validated abstinence rates were 15.7% (n = 64), 14.2% (n = 58), and 8.8% (n = 22) in the EC, varenicline, and NRT study arms, respectively. The quit rate in the EC arm was noninferior to the varenicline arm (absolute risk reduction, 1.47%; 95% CI, -1.41% to 4.34%) and higher than in the NRT arm (odds ratio, 1.92; 95% CI, 1.15-3.21). Treatment adherence was similar in all study arms during the initial 3 months, but 257 participants (62.8%) in the EC arm were still using ECs at 6 months, with no further use in the 2 other study arms. The most common adverse reactions were throat irritation (32 [7.8%]) and mouth irritation (28 [6.9%]) in the EC arm, nausea (36 [8.8%]) in the varenicline arm, and throat irritation (20 [8.0%]) and mouth irritation (22 [8.8%]) in the NRT arm. No serious adverse events were recorded. Conclusions and Relevance: The results of this randomized clinical trial found that when all treatments were provided with minimal behavior support, the efficacy of EC was noninferior to varenicline and superior to nicotine chewing gum. Trial Registration: Chinese Clinical Trial Registry: ChiCTR2100048156.

Oscillation of type IV pili regulated by the circadian clock in cyanobacterium Synechococcus elongatus PCC7942.

Type IV pili (TFP) are known to be functionally related to cell motilities and natural transformation in many bacteria. However, the molecular and ecological functions of the TFP have rarely been reported for photosynthetic cyanobacteria. Here, by labeling pili in model cyanobacterium Synechococcus elongatus PCC 7942 (Syn7942), we have quantitatively characterized the TFP and its driven twitching motility in situ at the single-cell level. We found an oscillating pattern of TFP in accordance with the light and dark periods during light-dark cycles, which is correlated positively to the oscillating pattern of the natural transformation efficiency. We further showed that the internal circadian clock plays an important role in regulating the oscillating pattern of TFP, which is also supported by evidences at the molecular level by tracking the expression of 16 TFP-related genes. This study adds a detailed picture toward the gap between TFP and its relations to circadian regulations in Syn7942.

Alleviative Effect of Lactoferrin Interventions Against the Hepatotoxicity Induced by Titanium Dioxide Nanoparticles.

The current study was designed to investigate the alleviative effect of lactoferrin interventions against the hepatotoxicity induced by titanium dioxide nanoparticles (TiO2-NPs). Thirty male Wistar rats were divided into six groups with 5 rats in each group. The first and second groups were intragastrically administered normal saline and TiO2-NPs (100 mg/kg body weight) as the negative control (NC) and TiO2-NP groups. The third, fourth, and fifth groups were intragastrically administered lactoferrin at concentrations of 100, 200, and 400 mg/kg body weight in addition to TiO2-NPs (100 mg/kg body weight). The sixth group was intragastrically administered Fuzheng Huayu (FZHY) capsules at a concentration of 4.6 g/kg body weight in addition to TiO2-NPs (100 mg/kg body weight) as the positive control group. After treatment for 4 weeks, the concentrations of lactoferrin were optimized based on the liver index and function results. Subsequently, the alleviative effects of lactoferrin interventions against TiO2-NP-induced hepatotoxicity in rat liver tissues, including the effects on histological damage, oxidative stress-related damage, inflammation, fibrosis, DNA damage, apoptosis, and gene expression, were investigated using histopathological, biochemical, and transcriptomic assays. The results showed that 200 mg/kg lactoferrin interventions for 4 weeks not only ameliorated the liver dysfunction and histopathological damage caused by TiO2-NP exposure but also inhibited the oxidative stress-related damage, inflammation, fibrosis, DNA damage, and apoptosis in the liver tissues of TiO2-NP-exposed rats. The transcriptomic results confirmed that the alleviative effect of lactoferrin interventions against the TiO2-NP exposure-induced hepatotoxicity was related to the activation of the PI3K/AKT signaling pathway.

Genetically encoded zinc-binding collagen-like protein hybrid hydrogels for wound repair.

Incorporating zinc oxide nanoparticles (ZnOnps) into collagen is a promising strategy for fabricating biomaterials with excellent antibacterial activity, but modifications are necessary due to the low zinc binding affinity of native collagen, which can cause disturbances to the functions of both ZnOnps and collagen and result in heterogeneous effects. To address this issue, we have developed a genetically encoded zinc-binding collagen-like protein, Zn-eCLP3, which was genetically modified by Scl2 collagen-like protein. Our study found that Zn-eCLP3 has a binding affinity for zinc that is 3-fold higher than that of commercialized type I collagen, as determined by isothermal titration calorimetry (ITC). Using ZnOnps-coordinated Zn-eCLP3 protein and xanthan gum, we prepared a hydrogel that showed significantly stronger antibacterial activity compared to a collagen hydrogel prepared in the same manner. In vitro cytocompatibility tests were conducted to assess the potential of the Zn-eCLP3 hydrogel for wound repair applications. In vivo experiments, which involved an S. aureus-infected mouse trauma model, showed that the application of the Zn-eCLP3 hydrogel resulted in rapid wound regeneration and increased expression of collagen-1α and cytokeratin-14. Our study highlights the potential of Zn-eCLP3 and the hybrid hydrogel for further studies and applications in wound repair.

Regulation of species metabolism in synthetic community systems by environmental pH oscillations.

Constructing a synthetic community system helps scientist understand the complex interactions among species in a community and its environment. Herein, a two-species community is constructed with species A (artificial cells encapsulating pH-responsive molecules and sucrose) and species B (Saccharomyces cerevisiae), which causes the environment to exhibit pH oscillation behaviour due to the generation and dissipation of CO2. In addition, a three-species community is constructed with species A' (artificial cells containing sucrose and G6P), species B, and species C (artificial cells containing NAD+ and G6PDH). The solution pH oscillation regulates the periodical release of G6P from species A'; G6P then enters species C to promote the metabolic reaction that converts NAD+ to NADH. The location of species A' and B determines the metabolism behaviour in species C in the spatially coded three-species communities with CA'B, CBA', and A'CB patterns. The proposed synthetic community system provides a foundation to construct a more complicated microecosystem.

The Dunning-Kruger effect: subjective health perceptions on smoking behavior among older Chinese adults.

BACKGROUND: The intrinsic damage and external hazards of smoking are major risk factors for poorer health and are recognized as a global health issue of concern in geriatric health. This study aims to assess the Dunning-Kruger effect through the influence of subjective health perceptions on smoking behavior in older adults. METHODS: This study used data from the 2018 Chinese Longitudinal Healthy Longevity Survey (N = 9,683) provided by the Center for Healthy Aging and Development Studies at Peking University. A binary logistic model was used to examine whether the Dunning-Kruger effect affects smoking behavior in older adults, and a linear probability model was used as a commentary baseline model for logistic regression to prevent measurement bias. In addition, a mediating analysis was used to examine the mechanisms through which the Dunning-Kruger effect occurs. RESULTS: Older adults often overestimated their current health status and underestimated the health risks of smoking, causing the Dunning-Kruger effect to arise from their inadequate self-perceived health (i.e., older adults are more likely to smoke when they have better self-rated health or when hypertension, cardiopathy, stroke, and diabetes have little or no impact on their daily lives). These observations can be explained by the older adults' subjective health perceptions arising from their ingenuous understanding of their health, which indirectly influences their smoking behavior to some extent. CONCLUSION: Older adults' self-perceived health was associated with smoking behavior. Public health institutions should improve older adults' health perceptions so that they objectively understand their own health status.

Genetically encoded in situ gelation redox-responsive collagen-like protein hydrogel for accelerating diabetic wound healing.

Genetically encoded collagen-like protein-based hydrogels have demonstrated remarkable efficacy in promoting the healing process in diabetic patients. However, the current methods for preparing these hydrogels pose significant challenges due to harsh reaction conditions and the reliance on chemical crosslinkers. In this study, we present a genetically encoded approach that allows for the creation of protein hydrogels without the need for chemical additives. Our design involves the genetic encoding of paired-cysteine residues at the C- and N-terminals of a meticulously engineered collagen-like recombination protein. The protein-based hydrogel undergoes a gel-sol transition in response to redox stimulation, achieving a gel-sol transition. We provide evidence that the co-incubation of the protein hydrogel with 3T3 cells not only enhances cell viability but also promotes cell migration. Moreover, the application of the protein hydrogel significantly accelerates the healing of diabetic wounds by upregulating the expression of collagen-1α (COL-1α) and Cytokeratin 14 (CK-14), while simultaneously reducing oxidant stress in the wound microenvironment. Our study highlights a straightforward strategy for the preparation of redox-responsive protein hydrogels, removing the need for additional chemical agents. Importantly, our findings underscore the potential of this hydrogel system for effectively treating diabetic wounds, offering a promising avenue for future therapeutic applications.

Efficacy of a porcine reproductive and respiratory syndrome virus 1 (PRRSV-1) natural recombinant against a heterologous PRRSV-1 isolate both clustered within the subgroup of BJEU06-1-like isolates.

Porcine reproductive and respiratory syndrome virus 1 (PRRSV-1) has been prevalent in more than 20 provinces of China. However, no PRRSV-1-specific vaccine is commercially available in China. To evaluate the feasibility of using a low virulent PRRSV-1 isolate against potential outbreaks caused by virulent Chinese PRRSV-1 isolates, here we evaluated the efficacy of a low virulent PRRSV-1 HLJB1 strain isolated in 2014 as live vaccine against a virulent PRRSV-1 SD1291 strain isolated in 2022. Genome-based phylogenetic analysis showed that both HLJB1 and SD1291 were grouped within BJEU06-1-like isolates. However, they shared only 85.27% genomic similarity. Piglet inoculation and challenge study showed that HLJB1 inoculation could reduce viremia but did not significantly alleviate clinical signs and tissue lesions. Virus neutralization test indicated that HLJB1 inoculation could induce homologous neutralizing antibodies (NAbs) but no heterologous NAbs at 42 dpi. In addition, flow cytometric analyses showed that no memory T follicular helper (Tfh) cells against SD1291 and SD1291-specific IFN-γ secreting cells were induced by HLJB1 pre-inoculation. These results supported that HLJB1 inoculation only provides partial cross-protection against SD1291 infection even though they are clustered within the same PRRSV-1 subgroup, which is closely related to the failure in conferring cross-protective adaptive immune responses.

Effects of Drought Stress on Non-Structural Carbohydrates in Different Organs of Cunninghamia lanceolata.

The Chinese fir Cunninghamia lanceolata (Lamb.) Hook. is an important timber conifer species in China. Much has been studied about Chinese fir, but the distribution of non-structural carbohydrates (NSCs) among different organs (needles, branch, stem, and roots) under drought stress remains poorly understood. In this study, we used one-year-old C. lanceolata plantlets to evaluate the effects of simulated drought under four water regimes, i.e., adequate water or control, light drought, moderate drought, and severe drought stress corresponding to 80%, 60%, 50%, and 40%, respectively of soil field maximum capacity on various NSCs in the needles, branch, stem and roots. The degree and duration of drought stress had significant effects on fructose, glucose, sucrose, soluble sugar, starch, and NSC content in various organs (p < 0.05). Fructose content increased in stem xylem, stem phloem, and leaves. Glucose and sucrose content declined in stem and branch xylem under light drought stress and moderate drought stress, and increased under severe drought stress conditions. Soluble sugars content declined, and starch content increased in leaf and branch phloem, but the latter could not compensate for soluble sugar consumption in the whole plant, and therefore, total NSCs decreased. Correlation analysis showed that a significant positive correlation existed in the soluble sugar content between leaves and roots, and between xylem and phloem in the stems and branches. Chinese fir appears to have different NSCs distribution strategies in response to drought stress, viz., allocating more soluble sugars to fine roots and increasing starch content in the needles, as well as ensuring osmosis to prevent xylem embolism. Our study may broaden the understanding of the various mechanisms that Chinese fir and other plants have to enhance their tolerance to drought stress.

Pressure-induced nonmonotonic cross-over of steady relaxation dynamics in a metallic glass.

Relaxation dynamics, as a key to understand glass formation and glassy properties, remains an elusive and challenging issue in condensed matter physics. In this work, in situ high-pressure synchrotron high-energy X-ray photon correlation spectroscopy has been developed to probe the atomic-scale relaxation dynamics of a cerium-based metallic glass during compression. Although the sample density continuously increases, the collective atomic motion initially slows down as generally expected and then counterintuitively accelerates with further compression (density increase), showing an unusual nonmonotonic pressure-induced steady relaxation dynamics cross-over at ~3 GPa. Furthermore, by combining in situ high-pressure synchrotron X-ray diffraction, the relaxation dynamics anomaly is evidenced to closely correlate with the dramatic changes in local atomic structures during compression, rather than monotonically scaling with either sample density or overall stress level. These findings could provide insight into relaxation dynamics and their relationship with local atomic structures of glasses.

Nephroprotective Effects of Selenium Nanoparticles Against Sodium Arsenite-Induced Damages.

Introduction: The potential effects of selenium nanoparticles (SeNPs) administration on arsenic exposure-mediated nephrotoxicity by alleviating fibrosis, inflammation, oxidative stress-related damage, and apoptosis remains more detailed investigations. Methods: After the synthesis of selenium nanoparticles (SeNPs) by sodium selenite (Na2SeO3) through a versatile and green procedure, the biosafety of SeNPs was assessed by assaying renal functions and inflammation in mice. Subsequently, nephroprotective effects of SeNPs against sodium arsenite (NaAsO2)-induced damages were confirmed by biochemical, molecular, and histopathological assays, including renal function, histological lesion, fibrosis, inflammation, oxidative stress-related damage, and apoptosis in mice renal tissues and renal tubular duct epithelial cells (HK2 cells). Results: The excellent biocompatibility and safety of SeNPs prepared in this study were confirmed by the non-significant differences in the renal functions and inflammation levels in mice between the negative control (NC) and 1 mg/kg SeNPs groups (p>0.05). The results of biochemical, molecular, and histopathological assays confirmed that daily administration of 1 mg/kg SeNPs for 4 weeks not only ameliorated renal dysfunctions and injuries caused by NaAsO2 exposure but also inhibited the fibrosis, inflammation, oxidative stress-related damage, and apoptosis in the renal tissues of NaAsO2-exposed mice. In addition, altered viability, inflammation, oxidative stress-related damage, and apoptosis in the NaAsO2-exposed HK2 cells were effectively reversed after 100 µg/mL SeNPs supplementation. Conclusion: Our findings authentically confirmed the biosafety and nephroprotective effects of SeNPs against NaAsO2 exposure-induced damages by alleviating inflammation, oxidative stress-related damage, and apoptosis.

Orthonasal and retronasal odor identification in patients with parosmia.

OBJECTIVE: To compare retronasal and orthonasal perception in parosmic COVID-19 patients, in order to determine whether COVID-19 has a differential effect on these functions. METHODS: Using the Sniffin Sticks test battery orthonasal function was examined for odor threshold, discrimination and identification. Retronasal function was assessed using 20 tasteless aromatized powders. Gustatory function was measured using the Taste Strips test. RESULTS: This study included 177 patients (127 women, 50 men; mean age 45 years), of whom 127 (72%) were hyposmic and 50 (28%) normosmic. Compared to patients without parosmia, parosmic patients performed worse in odor identification for both orthonasal (F = 4.94, p = 0.03) and retronasal tests (F = 11.95, p < 0.01). However, an interaction effect between route of odor identification (orthonasal or retronasal) and parosmia status was found (F = 4.67, p = 0.03): patients with parosmia had relatively lower retronasal scores than patients without parosmia. CONCLUSION: Our results suggest that COVID-19 may affect the olfactory mucosa differently along the anterior-posterior axis, thereby possibly contributing to the pathophysiology of parosmia. Patients with parosmia also exhibit a higher degree of impairment when odors are presented through the retronasal route during eating and drinking.

Important Roles and Formation of Atmospheric Organosulfates in Marine Organic Aerosols: Influence of Phytoplankton Emissions and Anthropogenic Pollutants.

Organosulfates (OSs) could be potentially important compounds in marine organic aerosols, while their formation in marine atmospheres is far from clear due to a lack of cruise observations. In this work, shipboard atmospheric observations were conducted over the Yellow Sea and Bohai Sea to investigate the abundance and formation of biogenic isoprene/monoterpene-OSs in marine aerosols. The quantified OSs and NOSs accounted for 0.04-6.9% of marine organic aerosols and were 0.07-2.2% of the non-sea-salt (nss) sulfate in terms of sulfur content. Isoprene-related (nitrooxy-)OSs occupied 27-87% of the total quantified OSs, following the abundance order of summer > autumn > spring or winter. This order was driven by the marine phytoplankton biomass and sea surface temperature (SST), which controlled the seawater and atmospheric isoprene concentration levels. Under the severe impacts of anthropogenic pollutants from the East Asia continent in winter, monoterpene nitrooxy-OSs, generated with NOx involved in, increased to 34.4 ± 35.5 ng/m3 and contributed 68% of the quantified (nitrooxy-)OSs. Our results highlight the notable roles of biogenic OSs in marine organic aerosols over regions with high biological activity and high SST. The formation of biogenic OSs and their roles in altering marine aerosol properties calls for elaboration through cruise observations in different marine environments.

Mineral Nutrient Uptake, Accumulation, and Distribution in Cunninghamia lanceolata in Response to Drought Stress.

Mineral accumulation in plants under drought stress is essential for drought tolerance. The distribution, survival, and growth of Chinese fir (Cunninghamia lanceolata (Lamb.) Hook.), an evergreen conifer, can be affected by climate change, particularly seasonal precipitation and drought. Hence, we designed a drought pot experiment, using 1-year-old Chinese fir plantlets, to evaluate drought effects under simulated mild drought, moderate drought, and severe drought, which corresponds to 60%, 50%, and 40% of soil field maximum moisture capacity, respectively. A treatment of 80% of soil field maximum moisture capacity was used as control. Effects of drought stress on mineral uptake, accumulation, and distribution in Chinese fir organs were determined under different drought stress regimes for 0-45 days. Severe drought stress significantly increased phosphorous (P) and potassium (K) uptake at 15, 30 and 45 days, respectively, within fine (diameter < 2 mm), moderate (diameter 2-5 mm), and large (diameter 5-10 mm) roots. Drought stress decreased magnesium (Mg) and manganese (Mn) uptake by fine roots and increased iron (Fe) uptake in fine and moderate roots but decreased Fe uptake in large roots. Severe drought stress increased P, K, calcium (Ca), Fe, sodium (Na), and aluminum (Al) accumulation in leaves after 45 days and increased Mg and Mn accumulation after 15 days. In stems, severe drought stress increased P, K, Ca, Fe, and Al in the phloem, and P, K, Mg, Na, and Al in the xylem. In branches, P, K, Ca, Fe, and Al concentrations increased in the phloem, and P, Mg, and Mn concentrations increased in the xylem under severe drought stress. Taken together, plants develop strategies to alleviate the adverse effects of drought stress, such as promoting the accumulation of P and K in most organs, regulating minerals concentration in the phloem and xylem, to prevent the occurrence of xylem embolism. The important roles of minerals in response to drought stress should be further evaluated.

[A cervical cancer tissue-derived decellularized extracellular matrix scaffold for cervical cancer tissue reconstruction in vitro].

OBJECTIVE: The prepare decellularized extracellular matrix (ECM) scaffold materials derived from human cervical carcinoma tissues for 3D culture of cervical carcinoma cells. METHODS: Fresh human cervical carcinoma tissues were treated with sodium lauryl ether sulfate (SLES) solution to prepare decellularized ECM scaffolds. The scaffolds were examined for ECM microstructure and residual contents of key ECM components (collagen, glycosaminoglycan, and elastin) and genetic materials by pathological staining and biochemical content analysis. In vitro 3D culture models were established by injecting cultured cervical cancer cells into the prepared ECM scaffolds. The cells in the recellularized scaffolds were compared with those in a conventional 2D culture system for cell behaviors including migration, proliferation and epithelial-mesenchymal transition (EMT) wsing HE staining, immunohistochemical staining and molecular biological technology analysis. Resistance to 5-fluorouracil (5-Fu) of the cells in the two culture systems was tested by analyzing the cell apoptosis rates via flow cytometry. RESULTS: SLES treatment effectively removed cells and genetic materials from human cervical carcinoma tissues but well preserved the microenvironment structure and biological activity of ECM. Compared with the 2D culture system, the 3D culture models significantly promoted proliferation, migration, EMT and 5-Fu resistance of human cervical cancer cells. CONCLUSION: The decellularized ECM scaffolds prepared using human cervical carcinoma tissues provide the basis for construction of in vitro 3D culture models for human cervical cancer cells.

Glutathione and selenium nanoparticles have a synergistic protective effect during cryopreservation of bull semen.

Introduction: In the present study, the synergistic protective effect of co-supplementation of glutathione (GSH) with selenium nanoparticles (SeNPs) on the cryopreservation efficiency of bull semen was analyzed. Methods: After collection, the ejaculates of Holstein bulls were subsequently diluted with a Tris extender buffer supplemented with different concentrations of SeNPs (0, 1, 2, and 4 µg/ml), followed by semen equilibration at 4°C and assessment of sperm viability and motility. Subsequently, the ejaculates of Holstein bulls were pooled, split into four equal groups, and diluted with a Tris extender buffer supplemented with basic extender (negative control group, NC group), 2 µg/ml SeNPs (SeNPs group), 4 mM GSH (GSH group), and 4 mM GSH plus 2 µg/ml SeNPs (GSH + SeNPs group). After cryopreservation, motility, viability, mitochondrial activity, plasma membrane integrity, acrosome integrity, concentration of malondialdehyde (MDA), superoxide dismutase (SOD), and catalase (CAT), and ability of frozen-thawed sperm cells to support in vitro embryonic development were evaluated. Results and discussion: No side effect of SeNPs concentrations applied in the current study on the motility and viability of equilibrated bull spermatozoa was found. Meanwhile, supplementation of SeNPs significantly promoted the motility and viability of equilibrated bull spermatozoa. Furthermore, the co-supplementation of GSH with SeNPs effectively protected bull spermatozoa from cryoinjury as expressed by promoting semen motility, viability, mitochondrial activity, plasma membrane integrity, and acrosome integrity. Finally, the enhanced antioxidant capacity and embryonic development potential in the frozen-thawed bull spermatozoa cryopreserved by co-supplementation of GSH with SeNPs further confirmed the synergistic protective effect of co-supplementation of GSH with SeNPs on the cryopreservation of bull semen.

Probing FeSi, a d-electron topological Kondo insulator candidate, with magnetic field, pressure, and microwaves.

Recently, evidence for a conducting surface state (CSS) below 19 K was reported for the correlated d-electron small gap semiconductor FeSi. In the work reported herein, the CSS and the bulk phase of FeSi were probed via electrical resistivity ρ measurements as a function of temperature T, magnetic field B to 60 T, and pressure P to 7.6 GPa, and by means of a magnetic field-modulated microwave spectroscopy (MFMMS) technique. The properties of FeSi were also compared with those of the Kondo insulator SmB6 to address the question of whether FeSi is a d-electron analogue of an f-electron Kondo insulator and, in addition, a "topological Kondo insulator" (TKI). The overall behavior of the magnetoresistance of FeSi at temperatures above and below the onset temperature TS = 19 K of the CSS is similar to that of SmB6. The two energy gaps, inferred from the ρ(T) data in the semiconducting regime, increase with pressure up to about 7 GPa, followed by a drop which coincides with a sharp suppression of TS. Several studies of ρ(T) under pressure on SmB6 reveal behavior similar to that of FeSi in which the two energy gaps vanish at a critical pressure near the pressure at which TS vanishes, although the energy gaps in SmB6 initially decrease with pressure, whereas in FeSi they increase with pressure. The MFMMS measurements showed a sharp feature at TS ≈ 19 K for FeSi, which could be due to ferromagnetic ordering of the CSS. However, no such feature was observed at TS ≈ 4.5 K for SmB6.