[Exogenous Spermidine Regulates Ryegrass Root System Response to Cd Stress and Its Transcriptome Analysis].

The application of exogenous growth-regulating substances is an effective technique to enhance plant stress tolerance. Here, a hydroponic experiment was conducted to investigate the effects of exogenous basal application of 0.1 mmol·L-1 spermidine (Spd) on both the physiology and molecular biology of ryegrass root systems under varying degrees (0, 5, and 10 mg·L-1) of cadmium (Cd) stress using ryegrass as the test plants. The results of physiological studies revealed that Cd stress significantly reduced the physiological functions of the ryegrass root system, whereas the addition of Spd effectively alleviated the negative effects caused by Cd. The most significant effect was on the root soluble protein content, which increased by 90.91% and 158.35% compared with 5 mg·L-1and 10 mg·L-1 Cd alone. Spd also inhibited the accumulation of oxidative stress products malondialdehyde (MDA) and hydrogen peroxide (H2O2) by increasing the ascorbic acid (ASA) and glutathione (GSH) content and peroxidase (POD) activity, whereas the effects on root activity and superoxide dismutase (SOD) activity were not significant. The results of molecular biology studies demonstrated that 10 mg·L-1 Cd stress caused differential expression of a large number of genes in ryegrass roots, and the number of differentially expressed genes, differential significance, and differential multiplicity were significantly reduced after the application of exogenous Spd. The most significant part of the GO enrichment analysis shifted from responding to organic cyclic compounds and aldehyde/ketone group transferase activity to responding to trivalent iron ions and 2'-deoxymugineic-acid 2'-dioxygenase activity. Single gene expression heat map analysis revealed that exogenous Spd upregulated the expression of genes encoding zinc-iron transporter protein and 2'-deoxymugineic-acid 2'-dioxygenase, which improved the uptake and utilization of iron by the root system. In conclusion, the application of certain concentrations of Spd could effectively regulate the response of ryegrass roots to Cd stress, enhance its tolerance physiology, and mitigate the toxic effects of Cd.

Evaluating the potential of nanofiltration membranes for removing ammonium, nitrate, and nitrite in drinking water sources.

Advanced drinking water treatment process using nanofiltration (NF) membranes has gained attention recently because it removes many challenging constituents in contaminated surface waters, such as dissolved organics and heavy metals. However, much literature has reported high variations and uncertainties of NF membranes for removing nitrogen compounds in the contaminated water-ammonium (NH4+), nitrates (NO3-), and nitrites (NO2-). This study aimed to identify the ability of commercial NF membranes to remove NH4+, NO2-, and NO3- and clarify the mechanisms underlying their transport through NF membranes. This was examined by evaluating their rejection by three commercial NF membranes using artificial and actual river waters under various conditions (variable permeate flux, temperature, pH, and ionic strength). Ammonium commonly showed the highest removal among the three nitrogen compounds, followed by nitrites and nitrates. Interestingly, ammonium removal varied considerably from 6% to 86%, depending on the membrane type and operating conditions. The results indicated that the selected nitrogen compounds (NH4+, NO2-, and NO3-) could be highly rejected depending on the clearance between their hydrated radius and the membrane's pore walls. Further, the rejection of the lowest molecular-weight nitrogen compound (NH4+) could be higher than NO2- and NO3- due to its highest energy barrier and larger hydrated radius. This study suggests that compliance with the drinking water regulations of NH4+, NO2-, and NO3- can be reliably achieved by selecting appropriate membrane types and predicting the range of their removal under various feed water quality and operating conditions.

Ionizable drug delivery systems for efficient and selective gene therapy.

Gene therapy has shown great potential to treat various diseases by repairing the abnormal gene function. However, a great challenge in bringing the nucleic acid formulations to the market is the safe and effective delivery to the specific tissues and cells. To be excited, the development of ionizable drug delivery systems (IDDSs) has promoted a great breakthrough as evidenced by the approval of the BNT162b2 vaccine for prevention of coronavirus disease 2019 (COVID-19) in 2021. Compared with conventional cationic gene vectors, IDDSs can decrease the toxicity of carriers to cell membranes, and increase cellular uptake and endosomal escape of nucleic acids by their unique pH-responsive structures. Despite the progress, there remain necessary requirements for designing more efficient IDDSs for precise gene therapy. Herein, we systematically classify the IDDSs and summarize the characteristics and advantages of IDDSs in order to explore the underlying design mechanisms. The delivery mechanisms and therapeutic applications of IDDSs are comprehensively reviewed for the delivery of pDNA and four kinds of RNA. In particular, organ selecting considerations and high-throughput screening are highlighted to explore efficiently multifunctional ionizable nanomaterials with superior gene delivery capacity. We anticipate providing references for researchers to rationally design more efficient and accurate targeted gene delivery systems in the future, and indicate ideas for developing next generation gene vectors.

Microenvironment-driven sequential ferroptosis, photodynamic therapy, and chemotherapy for targeted breast cancer therapy by a cancer-cell-membrane-coated nanoscale metal-organic framework.

Designing and developing nanomedicine based on the tumor microenvironment (TME) for effective cancer treatment is highly desirable. In this work, polyvinyl pyrrolidone (PVP) dispersed nanoscale metal-organic framework (NMOF) of Fe-TCPP (TCPP = tetrakis (4-carboxyphenyl) porphyrin) loaded with hypoxia-activable prodrug tirapazamine (TPZ) and coated by the cancer cell membrane (CM) is constructed (the formed nanocomposite denoted as PFTT@CM). Due to the functionalization with the homologous cancer cell membrane, PFTT@CM is camouflaged to evade the immune clearance and preferentially accumulates at the tumor site. Once internalized by cancer cells, PFTT@CM is activated by the TME through redox reaction and Fenton reaction between Fe3+ in nano-platform and endogenous glutathione (GSH) and hydrogen peroxide (H2O2) to promote GSH exhausting as well as •OH and O2 production, which triggers ferroptosis and dramatically enhances photodynamic therapy (PDT) efficacy. Subsequently, the PDT process mediated by TCPP and light would consume oxygen and aggravate tumor hypoxia to further activate the prodrug TPZ for cancer chemotherapy. As a consequence, the TME-driven PFTT@CM nano-platform not only demonstrated its TME modulation ability but also showed a sequential synergistic therapy, which eventually inhibited the cancer cell proliferation. This multimodal nano-platform is expected to shed light on the design of TME-activatable reaction to reinforce the synergistic therapeutic outcome and facilitate the development of effective cancer nanomedicine.

MRCKß links Dasm1 to actin rearrangements to promote dendrite development.

Proper dendrite morphogenesis and synapse formation are essential for neuronal development and function. Dasm1, a member of the immunoglobulin superfamily, is known to promote dendrite outgrowth and excitatory synapse maturation in vitro. However, the in vivo function of Dasm1 in neuronal development and the underlying mechanisms are not well understood. To learn more, Dasm1 knockout mice were constructed and employed to confirm that Dasm1 regulates dendrite arborization and spine formation in vivo. We performed a yeast two-hybrid screen using Dasm1, revealing MRCKß as a putative partner; additional lines of evidence confirmed this interaction and identified cytoplasmic proline-rich region (823-947 aa) of Dasm1 and MRCKß self-activated kinase domain (CC1, 410-744 aa) as necessary and sufficient for binding. Using co-immunoprecipitation assay, autophosphorylation assay, and BS3 cross-linking assay, we show that Dasm1 binding triggers a change in MRCKß's conformation and subsequent dimerization, resulting in autophosphorylation and activation. Activated MRCKß in turn phosphorylates a class 2 regulatory myosin light chain, which leads to enhanced actin rearrangement, causing the dendrite outgrowth and spine formation observed before. Removal of Dasm1 in mice leads to behavioral abnormalities. Together, these results reveal a crucial molecular pathway mediating cell surface and intracellular signaling communication to regulate actin dynamics and neuronal development in the mammalian brain.

Modification-bioremediation of copper, lead, and cadmium-contaminated soil by combined ryegrass (Lolium multiflorum Lam.) and Pseudomonas aeruginosa treatment.

The principal objective of this study was to investigate the strengthened remediation effect and relevant mechanism of P. aeruginosa on ryegrass (Lolium multiflorum Lam.) for soil contaminated by Cu-Pb-Cd compound heavy metals. The results showed that the complex heavy metals' contamination had remarkable inhibiting effect on the growth of plants (P < 0.01), and the biomass of ryegrass's stem and leaves declined by 28.2%, while that of roots decreased by 34.7% after 45 days. The inoculation of P. aeruginosa promoted the growth of ryegrass in polluted soil, in which the biomass recovered to the same level of that in normal plant; the activity of both catalase and urease in the soil also increased strikingly (by 29.3% and 75.7%, respectively); the ratio of residual heavy metals in the soil decreased, while the acid extractable heavy metals increased notably. Therefore, the absorption and accumulation of ryegrass to the heavy metals in soil were improved to some extent; the bioconcentration factor of Cu, Pb, and Cd in ryegrass increased by 35.9%, 55.6%, and 283.5%, respectively. The exterior microorganism allowed the accumulation of Cu, Pb, and Cd in shoots of ryegrass increasing remarkably, while in roots, only the accumulation of Pb increased by 16.3%, and that of both Cu and Cd decreased. Besides, in the P. aeruginosa-inoculated system, the transfer factor of Cu and Cd in plants increased strikingly, while that of Pb decreased.

Synergistic Chemotherapy for Breast Cancer and Breast Cancer Brain Metastases via Paclitaxel-Loaded Oleanolic Acid Nanoparticles.

Breast cancer is the most common type of cancer in women. About 12% of all women in the United States will be diagnosed with breast cancer over their lifetimes. At the same time, incidences of brain metastases (BMs) are increasing and represent an emerging health threat. However, there is no effective chemotherapy for breast cancer brain metastases (BCBMs), which is largely due to lack of efficient delivery of antitumor drugs or drug combinations to the brain. In this study, oleanolic acid (OA), a natural pentacyclic triterpenoid compound with excellent antitumor activity, was found to form nanoparticles (NPs) and efficiently penetrate the brain for BCBMs treatment. On the basis of these findings, we developed a synergistic combinatorial chemotherapeutic regimen by formulating paclitaxel (PTX) into OA NPs and demonstrated that the resulting PTX-OA NPs effectively inhibited primary breast cancer and BCBMs in mouse xenografts. Collectively, this study introduces a new direction to treat primary breast cancer and BCBMs through noninvasive combination chemotherapy.

Synergistic Platinum(II) Prodrug Nanoparticles for Enhanced Breast Cancer Therapy.

Chemotherapy still accounts for a large proportion of the treatments of tumors, but the drug resistance and side effects caused by long-term chemotherapy should not be underestimated. In this work, the drug combination strategy has been widely developed to overcome the side effects brought by the use of single drugs and improve the therapeutic effect. However, in clinical applications, the co-delivery of drugs is very difficult, and different in vivo kinetics due to different drug properties will lead to a decrease in efficacy. Thus, the design of novel antitumor therapeutic agents, including new platinum agents, represents an area in need of urgent attention. Our investigation implies a promising strategy for the design of a platinum prodrug to enhance the treatment of breast cancer. A dual-drug delivery nanoparticle was developed for enhanced treatment of breast cancer based on a two-into-one co-delivery strategy. Through the synergistic effect of released cisplatin hydrate and tolfenamic acid (COX-2 inhibitor) from the coordination prodrug, the tumor growth is significantly suppressed, and the survival time is greatly extended in breast tumor-bearing mice.

A Cell Membrane-Targeting Self-Delivery Chimeric Peptide for Enhanced Photodynamic Therapy and In Situ Therapeutic Feedback.

Nowadays, cell membrane-targeted therapy, which owns high antitumor efficacy by avoiding cell barriers, has received great attention. Here, a cell membrane-targeted self-delivery theranostic chimeric peptide CMP-PpIX is designed for simultaneously targeted photodynamic therapy (PDT) of tumor and real-time therapeutic feedback. Self-assembled CMP-PpIX nanoparticles can effectively accumulate in tumor by enhanced permeability and retention effect without additional vector. And this chimeric peptide CMP-PpIX has low background fluorescence, which is due to its relatively high intramolecular Förster resonance energy transfer (FRET) quenching efficiency between 5(6)-carboxyfluorescein (FAM) and 4-(dimethylaminoazo)-benzene-4-carboxylic acid (Dabcyl). More importantly, CMP-PpIX can be anchored on the tumor cell membrane for more than 8 h. Under irradiation, reactive oxygen species produced by CMP-PpIX directly damage cell membrane and rapidly induce apoptosis, which significantly improve the efficacy of PDT in vitro and in vivo. Then, peptide sequence Asp-Glu-Val-Asp (DEVD) is subsequently cleaved by activated caspase-3 and activated caspase-7, which separates the FAM and Dabcyl and terminates the FRET process. Therefore, fluorescence of FAM is recovered to monitor the expression of activated caspase-3 in vitro and in vivo to feedback real-time PDT therapeutic efficacy. In general, a novel cell membrane-targeted self-delivery theranostic chimeric peptide offers new promise for effective imaging-guided PDT.

Synthesis of Bioreducible Polycations with Controlled Topologies.

Bioreducible polycations, which possess disulfide linkages in the backbone, have emerged as promising nucleic acid delivery carriers due to their high stability in extracellular physiological condition and bioreduction-triggered release of the genetic material. Further benefits of bioreducible polycations include decreased cytotoxicity due to intracellular reducing environment in the cytoplasm that contains high levels of reducing molecules such as glutathione. Here, we describe the synthesis of bioreducible polycations with emphasis on methods to control their topology.

Safety and satisfaction of myopic small-incision lenticule extraction combined with monovision.

BACKGROUND: To investigate the safety and optical quality of small-incision lenticule extraction (SMILE) combined with monovision, and patient satisfaction with the procedure. METHODS: The present study assessed a non-random case series involving 60 eyes of 30 patients (mean age 45.53 ± 3.20 years [range 41 to 52 years]) treated bilaterally using the VisuMax 500 system (Carl Zeiss Meditec, Jena, Germany) between January and July 2016. The target refraction was plano for the distance eye, and between - 0.5 and - 1.75 diopters (D) for the near eye. Visual acuity, refraction errors, ocular aberrations, and satisfaction questionnaire scores were calculated 1 year after surgery. RESULTS: All surgeries were uneventful, with a mean safety index of 1.03 and 1.04 in dominant and nondominant eyes, respectively. Binocular uncorrected distance visual acuity of all patients was ≥20/32, while binocular uncorrected near visual acuity was ≥20/40 1 year postoperatively. Higher-order aberration (0.45 ± 0.14, 0.51 ± 0.15 µm), spherical (0.18 ± 0.15, 0.21 ± 0.14 µm) and coma aberration (0.31 ± 0.16, 0.27 ± 0.17 µm) were identical between dominant and nondominant eyes after surgery. The overall satisfaction rate was 86.7% (26/30), with large contributions from age (OR = 1.76 95% CI: 1.03-2.53; P = 0.036). Binocular uncorrected distance visual acuity was related to preoperative spherical diopter (r = - 0.500; P = 0.005). CONCLUSIONS: Monovision appears to be a safe and effective option for myopia patients with presbyopia who are considering the SMILE procedure. Patients with younger age were more satisfied with the procedure.

Cap morphology after small-incision lenticule extraction and its effects on intraocular scattering.

AIM: To investigate cap morphology after small-incision lenticule extraction (SMILE) and its effects on intraocular scattering. METHODS: Sixty-five eyes of 33 patients undergoing SMILE were enrolled. In addition to regular evaluation, Fourier-domain optical coherence tomography was used to investigate cap thickness at 1d, 1wk, 1 and 3mo postoperatively. The optical quality including modulation transfer function cutoff frequency, Strehl ratio, Optical Quality Analysis System (OQAS) values, and objective scattering index (OSI), were evaluated using OQAS™. RESULTS: Cap thickness decreased from 1d to 1wk (P<0.001), but remained higher than intended thickness of 120 µm after 3mo (P<0.001). Cap thickness in central area was thinner than that of in the paracentral and peripheral areas (P<0.0001). Total number of microdistortions decreased from 1d to 3mo (P<0.0001). Pearson analysis revealed a weak correlation between OSI and standard deviation of cap thickness at 1d and 1mo, as well as between range of cap thickness and OSI at 1mo. No correlation was found between microdistortion and OSI, but a negative correlation existed between microdistortion and range at 1d and 1moafter surgery. CONCLUSION: The corneal cap tends to be more accurate and regular with time lapse. Better cap morphology tends to contribute less intraocular scattering in the eyes undergoing SMILE.

Refractive Regression and Changes in Central Corneal Thickness Three Years after Laser-Assisted Subepithelial Keratectomy for High Myopia in Eyes with Thin Corneas: A Retrospective Study.

This retrospective study aimed to explore refractive regression and central corneal thickness (CCT) following laser-assisted subepithelial keratectomy (LASEK) performed for the correction of high myopia in eyes with thin corneas. Forty patients (19 male, 21 female; mean age, 28.5 years) representing 76 treated eyes were included. The mean optical zone was 6.10 ± 0.32 mm, and the mean ablation depth was 121.53 ± 15.48 µm. CCT was significantly greater three years after surgery than at three months after surgery (425.66 ± 15.44 vs. 385.20 ± 12.81, respectively; p<0.001). The mean change in CCT from three months to three years was 40.46 ± 14.02 µm. The SE at three years was greater than that at three months (p<0.001). Although there was refractive regression, these data suggest that LASEK may have utility in the correction of high myopia in eyes with thin corneas.

Self-assembling Janus dendritic polymer for gene delivery with low cytotoxicity and high gene transfection efficiency.

Polycations have high DNA condensing ability, low immunogenicity, and great adaptability, which make them promising for gene delivery. However, low transfection efficiency and inevitable toxicity are challenges of cationic polymers. In this study, we prepared a novel Janus dendritic polymer via a one-step Michael addition reaction of branched low-molecular-weight polyethylenimine (PEI, 1800 Da) and dendritic polythioether. The amphiphilic dendritic polymer can self-assemble into stable nanomicelles with high surface charge potential (+91.8 mV). The obtained nanomicelles can be used as a gene delivery vector, which exhibits a higher gene transfection efficiency and much lower cytotoxicity as compared with gold standard PEI (branched, 25 kDa).

Utilization of High-Fructose Corn Syrup for Biomass Production Containing High Levels of Docosahexaenoic Acid by a Newly Isolated Aurantiochytrium sp. YLH70.

High-fructose corn syrup (HFCS) is an agro-source product and has been the most commonly used substitute for sugar as sweetener in food industry due to its low price and high solution property. In this study, the F55 HFCS, rich in fructose and glucose, was first tested for biomass and docosahexaenoic acid productions as a mixed carbon source by a newly isolated Aurantiochytrium sp.YLH70. After the compositions of the HFCS media were optimized, the results showed that the HFCS with additions of metal ion and vitamin at low concentrations was suitable for biomass and docosahexaenoic acid productions and the metal ion and sea salt had the most significant effects on biomass production. During the 5-l fed-batch fermentation, total HFCS containing 180 g l(-1) reducing sugar was consumed and yields of biomass, lipid, and DHA could reach 78.5, 51, and 20.1 g l(-1), respectively, at 114 h. Meanwhile, the daily productivity and the reducing sugar conversion yield for docosahexaenoic acid were up to 4.23 g l(-1)day(-1) and 0.11 g g(-1). The fatty acid profile of Aurantiochytrium sp.YLH70 showed that 46.4% of total fatty acid was docosahexaenoic acid, suggesting that Aurantiochytrium sp.YLH70 was a promising DHA producer.

Evaluating the association between pathological myopia and SNPs in RASGRF1. ACTC1 and GJD2 genes at chromosome 15q14 and 15q25 in a Chinese population.

BACKGROUND: This study investigated the association of the 27 SNPs located in RASGRF1. GJD2, and ACTC1 genes with pathological myopia in a Chinese Han population. METHODS: Myopia patients were stratified according to whether they did (n = 274) or did not (n = 131) have myopic macular degeneration (MMD). The SNPbrowser software was used to identify specific SNPs for analysis and minimal allele frequency of >20%, and a pairwise r(2) < 0.85 were genotyped using MALDI-TOF mass spectrometry. RESULTS: Before controlling for false discovery rate, the frequency of the rs1867315 C/C genotype compared with healthy controls was lower in the myopia group (p = 0.006) and in myopia patients without macular degeneration (p = 0.019). The frequency of the rs670957A/A genotype was also lower in patients without MMD compared with controls (p = 0.045). For rs2070664, the frequency of the A allele was higher in the patients with MMD compared to those without MMD (p = 0.032). After controlling for a false discovery rate of 5%, there was no significant difference in genotype and allele frequencies between these groups. CONCLUSION: In this study, there was no association of the analyzed SNPs located in RASGRF1. GJD2, and ACTC1 with pathological myopia, suggesting that SNPs included in our study have no or a limited role in causing pathologic myopia in this Chinese Han population.

Biodegradable large compound vesicles with controlled size prepared via the self-assembly of branched polymers in nanodroplet templates.

Generally, it is very difficult to control the size of large compound vesicles. Here, we introduce a novel method for the preparation of biodegradable large compound vesicles with controlled size and narrow size distribution by using aqueous nanodroplets as templates.

Hemokinin-1(4-11)-induced analgesia selectively up-regulates δ-opioid receptor expression in mice.

Our previous studies have shown that an active fragment of human tachykinins (hHK-1(4-11)) produced an opioid-independent analgesia after intracerebroventricular (i.c.v.) injection in mice, which has been markedly enhanced by a δ OR antagonist, naltrindole hydrochloride (NTI). In this study, we have further characterized the in vivo analgesia after i.c.v. injection of hHK-1(4-11) in mouse model. Our qRT-PCR results showed that the mRNA levels of several ligands and receptors (e.g. PPT-A, PPT-C, KOR, PDYN and PENK) have not changed significantly. Furthermore, neither transcription nor expression of NK1 receptor, MOR and POMC have changed noticeably. In contrast, both mRNA and protein levels of DOR have been up-regulated significantly, indicating that the enhanced expression of δ opioid receptor negatively modulates the analgesia induced by i.c.v. injection of hHK-1(4-11). Additionally, the combinatorial data from our previous and present experiments strongly suggest that the discriminable distribution sites in the central nervous system between hHK-1(4-11) and r/mHK-1 may be attributed to their discriminable analgesic effects. Altogether, our findings will not only contribute to the understanding of the complicated mechanisms regarding the nociceptive modulation of hemokinin-1 as well as its active fragments at supraspinal level, but may also lead to novel pharmacological interventions.