Pulmonary delivery of mucus-traversing PF127-modified silk fibroin nanoparticles loading with quercetin for lung cancer therapy

The mucosal barrier remains a major barrier in the pulmonary drug delivery system,as mucociliary clearance in the airway accelerates the removal of inhaled nanoparticles(NPs).Herein,we designed and developed the inhalable Pluronic F127-modified silk fibroin NPs loading with quercetin(marked as QR-SF(PF127)NPs),aiming to solve the airway mucus barrier and improve the cancer therapeutic effect of QR.The PF127 coating on the SF NPs could attenuate the interaction between NPs and mucin proteins,thus facilitating the diffusion of SF(PF127)NPs in the mucus layer.The QR-SF(PF127)NPs had particle sizes of approximately 200 nm with negatively charged surfaces and showed constant drug release properties.Fluorescence recovery after photobleaching(FRAP)assay and transepithelial transport test showed that QR-SF(PF127)NPs exhibited superior mucus-penetrating ability in artificial mucus and monolayer Calu-3 cell model.Notably,a large amount of QR-SF(PF127)NPs distributed uniformly in the mice airway section,indicating the good retention of NPs in the respiratory tract.Themicemelanoma lungmetastasismodel was established,and the therapeutic effect of QR-SF(PF127)NPs was significantly improved in vivo.PF127-modified SF NPs may be a promising strategy to attenuate the interaction with mucin proteins and enhancemucus penetration efficiency in the pulmonary drug delivery system.

Growing Intact Membrane by Tuning Carbon Down to Ultrasmall 0.37 nm Microporous Structure for Confining Dissolution of Polysulfides Toward High-Performance Sodium–Sulfur Batteries

Room temperature sodium–sulfur(Na–S)batteries are severely hampered by dissolution of polysulfides into electrolytes.Herein,a facile approach is used to tune a biomass-derived carbon down to an ultrasmall 0.37 nm microporous structure for the first time as a cathode in sodium–sulfur batteries.This produced an intact uniform Na2S membrane to greatly confine the dissolution of polysulfides while realizing a direct solid phase conversion for complete reduction of sulfur to Na2S,which delivers a sulfur loading of 1 mg cm−2(50 wt.%),an excellent rate capacity(933 mAh g^(−1)@0.1 A g^(−1)and 410 mAh g^(−1)@2Ag^(−1)),long cycle performance(0.036%per cycle decay at 1 A g^(−1)after 1500 cycles),and a high energy density for 373 Wh kg^(−1)(0.1 A g^(−1))based on whole electrode weight(active sulfur loading+carbon),ranking the best among all reported plain carbon cathode-based room temperature sodium–sulfur batteries in terms of the cycle life and rate capacity.It is proposed that the solid Na2S produced in the ultrasmall pores(0.37 nm)can be squeezed out to grow an intact membrane on the electrode surface covering the outlet of the pores and greatly depressing the dissolution effect of polysulfides for the long cycle life.This work provides a green chemistry to recycle wastes for sustainable energies and sheds light on design of a unique pore structure to effectively block the dissolution of polysulfides for high-performance sodium–sulfur batteries.

Gas-propelled nanomotors alleviate colitis through the regulation of intestinal immunoenvironment-hematopexis-microbiota circuits

The progression of ulcerative colitis(UC)is associated with immunologic derangement,intestinal hemorrhage,and microbiota imbalance.While traditional medications mainly focus on mitigating inflammation,it remains challenging to address multiple symptoms.Here,a versatile gas-propelled nanomotor was constructed by mild fusion of post-ultrasonic CaO_(2) nanospheres with Cu_(2)O nanoblocks.The resulting CaO_(2)–Cu_(2)O possessed a desirable diameter(291.3 nm)and a uniform size distribution.It could be efficiently internalized by colonic epithelial cells and macrophages,scavenge intracellular reactive oxygen/nitrogen species,and alleviate immune reactions by pro-polarizing macrophages to the anti-inflammatory M2 phenotype.This nanomotor was found to penetrate through the mucus barrier and accumulate in the colitis mucosa due to the driving force of the generated oxygen bubbles.Rectal administration of CaO_(2)–Cu_(2)O could stanch the bleeding,repair the disrupted colonic epithelial layer,and reduce the inflammatory responses through its interaction with the genes relevant to blood coagulation,anti-oxidation,wound healing,and anti-inflammation.Impressively,it restored intestinal microbiota balance by elevating the proportions of beneficial bacteria(e.g.,Odoribacter and Bifidobacterium)and decreasing the abundances of harmful bacteria(e.g.,Prevotellaceae and Helicobacter).Our gas-driven CaO_(2)–Cu_(2)O offers a promising therapeutic platform for robust treatment of UC via the rectal route.

蚕肠道好氧微生物菌群的研究

以夏芳蚕品种为材料 ,对其不同蚕期、不同龄期间肠道好氧微生物种群构成及其变化情况进行了系统的研究。结果表明 :春蚕肠道好氧菌数量较多 ,以肠杆菌、微球菌科菌和芽胞杆菌科菌为主 ;秋蚕肠道好氧菌只有少量微球菌科菌和芽胞杆菌科菌存在 ;人工饲料育肠道好氧菌数量较少 。

Multiple Interval Mapping for Whole Cocoon Weight and Related Economically Important Traits QTL in Silkworm(Bombyx mori)

A backcrossed population （BC1） derived from a cross between C100 and Dazao was obtained. The quantitative trait loci （QTLs） of the economically important traits for whole cocoon weight, cocoon shell weight, ratio of cocoon shell and weight of pupae, etc., were analyzed for the first time using the multiple interval mapping software WinQTLCart2.0. In total 40 QTLs were detected and contributed to 21 groups based on the constructed linkage map. According to the mapping results, 2, 2, 3, and 2 major QTLs explained over 20% of total phenotypic variations, whereas four QTLs, namely qCW-19, qSW-2, qCSR-4, and qPW-23, explained more than 30% of total phenotypic variations for whole cocoon weight, cocoon shell weight, ratio of cocoon shell and weight of pupae, respectively. Correlated traits QTLs often share the same location. Furthermore, most of the detected QTLs were closed to one-side marker. By using the very closed markers, positive QTLs can be aggregated, which can form a basis for molecular marker-assisted selection and breeding.

A secreted phospholipase A_(2) (BmsPLA_(2)) regulates melanization of immunity through BmDDC in the silkworm Bombyx mori

Insect immune-associated phospholipase A_(2) (PLA_(2)) is an important target of pathogen invasion. Melanization, an effective defense response, has significant correlations with other immune responses to coordinate immune attack against invaders. However, the effect of PLA_(2) on melanization has not yet been reported in insects or other arthropods. In this work, we cloned a PLA_(2) gene (BmsPLA_(2)), and its protein had characteristic features of secreted PLA_(2) (sPLA_(2)). After injection of bacteria, BmsPLA_(2) expression and sPLA_(2) activity in hemolymph significantly increased. BmsPLA_(2) fluorescence was transferred from the cytoplasm to the cell membranes of circulating hemocytes. These results indicated that BmsPLA_(2) was related to hemolymph immunity in silkworms. Interestingly, reducing BmsPLA_(2) by RNA interference decreased melanosis (melanistic hemocytes) levels in vivo and in vitro, while BmsPLA_(2) overexpression had the opposite effect. The larval survival and melanization rate in the hemocoel both slowed depending on the PLA_(2) inhibitor dosage. These results demonstrated that BmsPLA_(2) plays a role in melanization during the immune process of silkworms. Surprisingly, the level of BmDDC matched the degree of melanization in various observations. BmDDC expression showed a significant increase, with the peak occurring later than that of BmsPLA_(2) after injection of bacteria, implying that BmsPLA_(2) was activated prior to BmDDC. Moreover, the alteration of BmsPLA_(2) by RNA interference or overexpression led to altered BmDDC levels. These results suggested that BmsPLA_(2) regulates the melanization response in silkworms through BmDDC. Our study proposes a new regulatory mechanism of the melanization response and new directions for understanding the complex immune networks of insects.

Bioinspired Hemostatic Strategy via Pulse Ejections for Severe Bleeding Wounds

Efficient hemostasis during emergency trauma with massive bleeding remains a critical challenge in prehospital settings.Thus,multiple hemostatic strategies are critical for treating large bleeding wounds.In this study,inspired by bombardier beetles to eject toxic spray for defense,a shape-memory aerogel with an aligned microchannel structure was proposed,employing thrombin-carrying microparticles loaded as a built-in engine to generate pulse ejections for enhanced drug permeation.Bioinspired aerogels,after contact with blood,can rapidly expand inside the wound,offering robust physical barrier blocking,sealing the bleeding wound,and generating a spontaneous local chemical reaction causing an explosivelike generation of CO_(2) microbubbles,which provide propulsion thrust to accelerate burst ejection from arrays of microchannels for deeper and faster drug diffusion.The ejection behavior,drug release kinetics,and permeation capacity were evaluated using a theoretical model and experimentally demonstrated.This novel aerogel showed remarkable hemostatic performance in severely bleeding wounds in a swine model and demonstrated good degradability and biocompatibility,displaying great potential for clinical application in humans.

Mulberry Biomass-Derived Nanomedicines Mitigate Colitis through Improved Inflamed Mucosa Accumulation and Intestinal Microenvironment Modulation

The therapeutic outcomes of conventional oral medications against ulcerative colitis(UC)are restricted by inefficient drug delivery to the colitis mucosa and weak capacity to modulate the inflammatory microenvironment.Herein,a fluorinated pluronic(FP127)was synthesized and employed to functionalize the surface of mulberry leaf-derived nanoparticles(MLNs)loading with resveratrol nanocrystals(RNs).The obtained FP127@RN-MLNs possessed exosome-like morphologies,desirable particle sizes(around 171.4 nm),and negatively charged surfaces(−14.8 mV).The introduction of FP127 to RN-MLNs greatly improved their stability in the colon and promoted their mucus infiltration and mucosal penetration capacities due to the unique fluorine effect.

Analysis of cytochrome P450 genes in silkworm genome (Bombyx mori)

We have searched the Bombyx mori genome for members of the major enzyme family, the Cytochrome P450s, which carry out multiple reactions to enable organisms to rid themselves of foreign compounds. As a result, 86 putative P450s were discovered in silkworm genome, which are thought to belong to 32 subfamilies. A comparative genomic analysis with Drosophila melanogaster reveals that the two insects have some similar P450 distribution pat-terns but still have some obvious differences. Especially, the diverse distribution exists in 7 p450 subfamilies, which are CYP4A, CYP4C, CYP4D, CYP6A, CYP6AE, CYP6B and CYP9A. Fur-thermore, we collected expression sequence tag (EST) evidence for 49 putative P450s genes, which are expressed at the transcriptional level and more likely to be true P450s.

Chromosome-level Genomes Reveal the Genetic Basis of Descending Dysploidy and Sex Determination in Morus Plants

Multiple plant lineages have independently evolved sex chromosomes and variable karyotypes to maintain their sessile lifestyles through constant biological innovation.Morus notabilis,a dioecious mulberry species,has the fewest chromosomes among Morus spp.,but the genetic basis of sex determination and karyotype evolution in this species has not been identified.In this study,three high-quality genome assemblies were generated for Morus spp.[including dioecious M.notabilis(male and female)and Morus yunnanensis(female)]with genome sizes of 301–329 Mb and were grouped into six pseudochromosomes.Using a combination of genomic approaches,we found that the putative ancestral karyotype of Morus species was close to 14 protochromosomes,and that several chromosome fusion events resulted in descending dysploidy(2n=2x=12).We also characterized a~6.2-Mb sex-determining region on chromosome 3.Four potential male-specific genes,a partially duplicated DNA helicase gene(named MSDH)and three Ty3_Gypsy long terminal repeat retrotransposons(named MSTG1/2/3),were identified in the Y-linked area and considered to be strong candidate genes for sex determination or differentiation.Population genomic analysis showed that Guangdong accessions in China were genetically similar to Japanese accessions of mulberry.In addition,genomic areas containing selective sweeps that distinguish domesticated mulberry from wild populations in terms of flowering and disease resistance were identified.Our study provides an important genetic resource for sex identification research and molecular breeding in mulberry.

Fabrication of Sulfated Silk Fibroin‑Based Blend Nanofbrous Membranes for Lysozyme Adsorption

In this work,the silk fbroin/cellulose blend nanofbrous membranes modifed with sodium-3-sulfobenzoate(S-SCBNM)were fabricated by electrospinning technique for lysozyme adsorption.The morphology and structure of membranes was observed.The efect of sulfate contents,pH values and concentration of lysozyme on adsorption performance were studied,respectively.The reuse capacity was evaluated.The results showed that the resultant S-SCBNM exhibited integrated properties of ultrathin fber diameter(148 nm),fast adsorption equilibrium,excellent adsorption performance(636 mg g−1)and good reversibility.We envision that this new class of green and natural blend membranes is particularly promising for the development of proteins separation.

Microcluster colloidosomes for hemostat delivery into complex wounds:A platform inspired by the attack action of torpedoes

Complex yet lethal wounds with uncontrollable bleeding hinder conventional hemostats from clotting blood at the source or deep sites of injury vasculature,thereby causing massive blood loss and significantly increased mortality.Inspired by the attack action of torpedoes,we synthesized microcluster(MC)colloidosomes equipped with magnetic-mediated navigation and“blast”systems to deliver hemostats into the cavity of vase-type wounds.CaCO_(3)/Fe_(2)O_(3)(CF)microparticles functionalized with Arg-Gly-Asp(RGD)modified polyelectrolyte multilayers were co-assembled with oppositely charged zwitterionic carbon dots(CDs)to form MC colloidosomes,which were loaded with thrombin and protonated tranexamic acid(TXA-NH_(3)^(+)).The composite microparticles moved against blood flow under magnetic mediation and simultaneously disassembled for the burst release of thrombin stimulated by TXA-NH_(3)^(+).The CO_(2) bubbles generated during disassembly produced a“blast”that propelled thrombin into the wound cavity.Severe bleeding in a vase-type hemorrhage model in the rabbit liver was rapidly controlled within~60 s.Furthermore,in vivo subcutaneous muscle and liver implantation models demonstrated excellent biodegradability of MC colloidosomes.This study is the first to propose a novel strategy based on the principle of torpedoes for transporting hemostats into vase-type wounds to achieve rapid hemostasis,creating a new paradigm for combating trauma treatment.

Erythrocyte membrane-camouflaged nanoworms with on-demand antibiotic release for eradicating biofilms using near-infrared irradiation

The increase in the number of resistant bacteria caused by the abuse of antibiotics and the emergence of biofilms significantly reduce the effectiveness of antibiotics.Bacterial infections are detrimental to our life and health.To reduce the abuse of antibiotics and treat biofilm-related bacterial infections,a biomimetic nano-antibacterial system,RBCM-NW-G namely,that controls the release of antibiotics through near infrared was prepared.The hollow porous structure and the high surface activity of nanoworms are used to realize antibiotic loading,and then,biomimetics are applied with red blood cell membranes(RBCM).RBCM-NW-G,which retains the performance of RBCM,shows enhanced permeability and retention effects.Fluorescence imaging in mice showed the effective accumulation of RBCM-NW-G at the site of infection.In addition,the biomimetic nanoparticles showed a longer blood circulation time and good biocompatibility.Anti-biofilm test results showed damage to biofilms due to a photothermal effect and a highly efficient antibacterial performance under the synergy of the photothermal effect,silver iron,and antibiotics.Finally,by constructing a mouse infection model,the great potential of RBCM-NW-G in the treatment of in vivo infections was confirmed.

Integrin β2 and β3:Two plasmatocyte markers deepen our understanding of the development of plasmatocytes in the silkworm Bombyx mori

Insect hemocytes play important biological roles at developmental stages,metamorphosis,and innate immunity.As one of the most abundant cell types,plasmatocytes can participate in various innate immune responses,especially in encapsulation and node formation.Here,2 molecular markers of plasmatocytes,consisting of integrinβ2 andβ3,were identified and used to understand the development of plasmatocytes.Plasmatocytes are widely distributed in the hematopoietic system,including circulating hemolymph and hematopoietic organs(HPOs).HPOs constantly release plasmatocytes with high proliferative activity in vitro;removal of HPOs leads to a dramatic reduction in the circulating plasmatocytes,and the remaining plasmatocytes gradually lose their ability to proliferate in vivo.Our results demonstrated that the release of plasmatocytes from HPOs is regulated by insulin-mediated signals and their downstream pathways,including PI3K/Akt and MAPK/Erk signals.The insulin/PI3K/Akt signaling pathway can significantly irritate the hematopoiesis,and its inhibitor LY294002 could inhibit the hemocytes discharged from HPOs.While the insulin/MAPK/Erk signaling pathway plays a negative regulatory role,inhibiting its activity with U0126 can markedly promote the discharge of plasmatocytes from HPOs.Our results indicate that the circulating plasmatocytes are mainly generated and discharged by HPOs.This process is co-regulated by the PI3K/Akt and MAPK/Erk signals in an antagonistic manner to adjust the dynamic balance of the hemocytes.These findings can enhance our understanding of insect hematopoiesis.

Bmelo12,an elongase of very long-chain fatty acids gene,regulates silk yield in Bombyx mori

In silkworm,cocoon shell weight(CSW),an index of silk yield,is a quantitative trait(Nagaraju and Goldsmith,2002),and multiple quantitative trait loci(QTLs)located on different chromosomes have been found(Zhan et al.,2009;Zhang et al.,2010;Li et al.,2015;Fang et al.,2020).

Gemini Dressing with Both Super‑hydrophilicity and ‑hydrophobicity Pursuing Isolation of Blood Cells for Hemostasis and Wound Healing

Achieving efficient hemostasis and wound management is vital to preserve life and restore health in case of extensive hemor-rhagic skin damage.Here,we develop a filter pump-like hierarchical porous-structure(HPS)dressing based on a non-woven substrate,konjac glucomannan(KGM)aerogel,and bi-functional microporous starch(BMS).The KGM aerogel intercalates into the non-woven network structure,forming a hydrophilic frame to stimulate the plasma permeation toward the interior in synergy with the hydrophilic pores of the BMS.The BMS surface forms a hydrophobic matrix that fills the spaces of the KGM hydrophilic frame,contributing to the isolation and aggregation of blood cells on the surface of the HPS dressing to establish rapid hemostasis.Animal model experiments suggest reliable HPS dressing hemostatic capacity,as it is able to stop ear artery and liver bleeding within 97.6±15.2 s and 67.8±5.4 s,respectively.Furthermore,the dressings exhibit antibacterial properties and enabled wound healing within 2 weeks.In vitro hemolysis and cytotoxicity tests also confirm the biocompatibility of HPS dressings.This novel“two-in-one”hemostatic dressing facilitates tissue repair of bleeding wounds over the entire recovery period,thereby providing a convenient strategy for wound management.

Natural exosome-like nanovesicles from edible tea flowers suppress metastatic breast cancer via ROS generation and microbiota modulation

Although several artificial nanotherapeutics have been approved for practical treatment of metastatic breast cancer,their inefficient therapeutic outcomes,serious adverse effects,and high cost of mass production remain crucial challenges.Herein,we developed an alternative strategy to specifically trigger apoptosis of breast tumors and inhibit their lung metastasis by using natural nanovehicles from tea flowers(TFENs).These nanovehicles had desirable particle sizes(131 nm),exosome-like morphology,and negative zeta potentials.Furthermore,TFENs were found to contain large amounts of polyphenols,flavonoids,functional proteins,and lipids.Cell experiments revealed that TFENs showed strong cytotoxicities against cancer cells due to the stimulation of reactive oxygen species(ROS)amplification.The increased intracellular ROS amounts could not only trigger mitochondrial damage,but also arrest cell cycle,resulting in the in vitro anti-proliferation,anti-migration,and anti-invasion activities against breast cancer cells.Further mice investigations demonstrated that TFENs after intravenous(i.v.)injection or oral administration could accumulate in breast tumors and lung metastatic sites,inhibit the growth and metastasis of breast cancer,and modulate gut microbiota.This study brings new insights to the green production of natural exosome-like nanoplatform for the inhibition of breast cancer and its lung metastasis via i.v.and oral routes.

Multi-responsive nanotheranostics with enhanced tumor penetration and oxygen self-producing capacities for multimodal synergistic cancer therapy

Incorporation of multiple functions into one nanoplatform can improve cancer diagnostic efficacy and enhance anti-cancer outcomes. Here, we constructed doxorubicin(DOX)-loaded silk fibroinbased nanoparticles(NPs) with surface functionalization by photosensitizer(N770). The obtained nanotheranostics(N770-DOX@NPs) had desirable particle size(157 nm) and negative surface charge(-25 m V). These NPs presented excellent oxygen-generating capacity and responded to a quadruple of stimuli(acidic solution, reactive oxygen species, glutathione, and hyperthermia). Surface functionalization of DOX@NPs with N770 could endow them with active internalization by cancerous cell lines, but not by normal cells. Furthermore, the intracellular NPs were found to be preferentially retained in mitochondria, which were also efficient for near-infrared(NIR) fluorescence imaging, photothermal imaging,and photoacoustic imaging. Meanwhile, DOX could spontaneously accumulate in the nucleus. Importantly, a mouse test group treated with N770-DOX@NPs plus NIR irradiation achieved the best tumorretardation effect among all treatment groups based on tumor-bearing mouse models and a patientderived xenograft model, demonstrating the unprecedented therapeutic effects of trimodal imagingguided mitochondrial phototherapy(photothermal therapy and photodynamic therapy) and chemotherapy.Therefore, the present study brings new insight into the exploitation of an easy-to-use, versatile, and robust nanoplatform for programmable targeting, imaging, and applying synergistic therapy to tumors.

Bombyx mori U-shaped regulates the melanization cascade and immune response via binding with the Lozenge protein

Zinc finger protein,an important transcription factor,regulates gene expression associated with various physiological and pathological processes.U-shaped,belong to the Friend of GATA(FOG)transcription factor,plays a crucial role in hematopoiesis by interacting with the GATA transcription factor as a co-factor.However,little is known about its functions in insects.In the present study,a U-shaped cDNA was identified and characterized from the silkworm Bombyx mori and its potential roles in innate immunity investigated.The predicted silkworm U-shaped amino acid sequence contained a classical nuclear localization signal(NLS)motif“GESSPKRRRR”at position 45CU459,and arginine residues at position 456 and 478 are the critical sites of the NLS.U-shaped mRNA was detected in all tested tissues of the B.mori;however,the highest levels were found in the hemocytes.U-shaped mRNA expression levels were upregulated in the hemocyte after the Escherichia coli and Staphylococcus aureus challenge.Furthermore,U-shaped knockdown significantly reduced the melanization process and suppressed the expression of melanization-associated genes,including PPO1,PPO2,PPAE and BAEE.In addition,U-shaped interacts with Lozenge protein to regulate the innate immune response of the insect.Our results revealed that U-shaped binds directly to Lozenge protein to modulate the melanization process and innate immune responses in silkworm.

Magnetic field-mediated Janus particles with sustained driving capability for severe bleeding control in perforating and inflected wounds

Severe bleeding in perforating and inflected wounds with forky cavities or fine voids encountered during prehospital treatments and surgical procedures is a complex challenge.Therefore,we present a novel hemostatic strategy based on magnetic field-mediated guidance.The biphasic Janus magnetic particle(MSS@Fe2O3-T)comprised aggregates ofα-Fe2O3 nanoparticles(Fe_(2)O_(3) NPs)as the motion actuator,negatively modified microporous starch(MSS)as the base hemostatic substrate,and thrombin as the loaded hemostatic drug.Before application,the particles were first wrapped using NaHCO_(3) and then doped with protonated tranexamic acid(TXA-NH_(3)^(+)),which ensured their high self-dispersibility in liquids.During application,the particles promptly self-diffused in blood by bubble propulsion and travelled to deep bleeding sites against reverse rushing blood flow under magnetic guidance.In vivo tests confirmed the superior hemostatic performance of the particles in perforating and inflected wounds(“V”-shaped femoral artery and“J”-shaped liver bleeding models).The present strategy,for the first time,extends the range of magnetically guided drug carriers to address the challenges in the hemorrhage control of perforating and inflected wounds.