Tetrodotoxin (TTX) is a potent neurotoxin with therapeutic potential, particularly in the fields of analgesia, cancer pain treatment, and drug addiction therapy. However, its high toxicity and lack of antidotes limit its clinical application. This study evaluated the acute toxicity and sub-acute toxicity of TTX via intramuscular injection in Sprague–Dawley (SD) rats. The acute toxicity test employed the Bliss method to determine the median lethal dose (LD50), with 10 rats per group (sex-balanced). Rats received a single intramuscular injection at doses of 8.2–20.0 μg/kg. The sub-acute toxicity study was conducted through daily intramuscular injections at doses of 1.5, 3.0, and 6.0 μg/kg/day for 28 consecutive days. Parameters, including body weight, food consumption, hematology, serum biochemistry, urinalysis, and histopathology, were assessed. Acute toxicity was characterized by squinting, reduced spontaneous activity, convulsions, hind limb rigidity, limb weakness, and mortality. In the sub-acute toxicity study, no toxicity-related changes associated with TTX treatment were observed in any dose group compared to controls, including in food consumption, hematology, urinalysis, and histopathological examination. Although statistically significant differences in body weight, serum biochemical, and organ weights were noted at certain time points in individual dose groups, these were not dose-dependent and were of minimal magnitude. The LD50 of TTX administered via a single intramuscular injection in SD rats was 13.1 μg/kg. In the sub-acute toxicity study, no adverse effects were observed up to the highest tested dose (6.0 μg/kg/day), which was therefore considered the no observed adverse effect level (NOAEL) under the conditions.

Cuproptosis, a novel approach utilizing copper carriers to trigger programmed cell death, exhibits promise for enhancing traditional therapies and activating robust adaptive immune responses. However, the uncontrolled release of Cu ions risks triggering cuproptosis in healthy tissues, potentially causing irreversible damage. To address this, we report on the use of a Cu-MOF (copper metal-organic framework) protective layer to regulate the biodegradation of copper-based nanomaterials. In situ formation of Cu-MOF on Cu2O nanocubes not only stabilizes the material under physiological conditions but also enhances its sonodynamic therapy (SDT) capabilities by establishing a Z-Scheme heterojunction. Upon SDT activation, the targeted Cu ion release at the tumor site triggers a cascade of reactions, generating reactive oxygen species (ROS) via Fenton-like processes and depleting glutathione (GSH). This ROS surge, combined with effective cuproptosis, modulates the immunosuppressive tumor microenvironment, inducing immunogenic cell death to eliminate primary tumors and inhibit metastasis. This study offers a new paradigm for the controlled integration of SDT, chemodynamic therapy (CDT), cuproptosis, and immunotherapy, achieving precise tumor-targeted treatment via controlled copper nanomaterial degradation.

Okadaic acid (OA), a diarrhetic shellfish toxin, causes significant intestinal and systemic toxicity. Currently, there are no approved therapeutics specifically targeting OA-induced diarrhea. Using high-throughput screening of 2131 natural compounds, we identified triptolide as a compound conferring protection against OA-induced apoptosis. In mice exposed to OA, triptolide administration reduced intestinal permeability and ameliorated mucosal injury. In HUVECs, triptolide (0.1-3 μM) dose-dependently increased cell viability and suppressed apoptotic activity. Biotin-based labeling identified the hydroxyl group as the essential active site of triptolide. RNA sequencing analysis showed that triptolide inhibited the OA-induced upregulation of endoplasmic reticulum (ER) protein synthesis-related genes. Furthermore, triptolide prevented the OA-induced decrease in the p-eIF-2α/eIF-2α ratio, thereby attenuating ER stress. These results demonstrate that triptolide mitigates OA-induced toxicity by inhibiting ER stress-mediated endothelial cell apoptosis and maintaining intestinal barrier integrity, highlighting its potential as a therapeutic agent against OA-related poisoning.

Progress has been made in generating spinal cord and trunk derivatives from neuromesodermal progenitors (NMPs). However, maintaining the self-renewal of NMPs in vitro remains a challenge. In this study, we developed a cocktail of small molecules and growth factors that induces human embryonic stem cells to produce self-renewing NMPs (srNMPs) under chemically defined conditions. These srNMPs maintain the state of neuromesodermal progenitors in prolonged culture and have the potential to generate mesodermal cells and neurons, even at the single-cell level. Additionally, suspended srNMP aggregates can spontaneously differentiate into all tissue types of early embryonic trunks. Furthermore, transplanted srNMP-derived muscle satellite cells or progenitors of motor neurons were integrated into skeletal muscle or the spinal cord, respectively, and contributed to regeneration in mouse models. In summary, srNMPs hold great promise for applications in developmental biology and as renewable cell sources for cell therapy for trunk and spinal cord injuries.

Photothermal therapy (PTT) of nanomaterials is an emerging novel therapeutic strategy for breast cancer. However, there exists an urgent need for appropriate strategies to enhance the antitumor efficacy of PTT and minimize damage to surrounding normal tissues. Piezo1 might be a promising novel photothermal therapeutic target for breast cancer. This study aims to explore the potential role of Piezo1 activation in the hyperthermia therapy of breast cancer cells and investigate the underlying mechanisms. Results showed that the specific agonist of Piezo1 ion channel (Yoda1) aggravated the cell death of breast cancer cells triggered by heat stress in vitro. Reactive oxygen species (ROS) production was significantly increased following heat stress, and Yoda1 exacerbated the rise in ROS release. GSK2795039, an inhibitor of NADPH oxidase 2 (NOX2), reversed the Yoda1-mediated aggravation of cellular injury and ROS generation after heat stress. The in vivo experiments demonstrate the well photothermal conversion efficiency of TiCN under the 1,064 nm laser irradiation, and Yoda1 increases the sensitivity of breast tumors to PTT in the presence of TiCN. Our study reveals that Piezo1 activation might serve as a photothermal sensitizer for PTT, which may develop as a promising therapeutic strategy for breast cancer.

CoSnO 3 nanocubes were successfully synthesized to achieve complete tumor eradication through mild NIR-II photothermally enhanced nanocatalytic cancer therapy.

Multiwalled carbon nanotubes (MWCNTs) have numerous applications in the field of carbon nanomaterials. However, the associated toxicity concerns have increased significantly because of their widespread use. The inhalation of MWCNTs can lead to nanoparticle deposition in the lung tissue, causing inflammation and health risks. In this study, celastrol, a natural plant medicine with potent anti-inflammatory properties, effectively reduced the number of inflammatory cells, including white blood cells, neutrophils, and lymphocytes, and levels of inflammatory cytokines, such as IL-1β, IL-6, and TNF-α, in mice lungs exposed to MWCNTs. Moreover, celastrol inhibited the activation of the NF-κB-signaling pathway. This study confirmed these findings by demonstrating comparable reductions in inflammation upon exposure to MWCNTs in mice with the deletion of NF-κB (P50-/-). These results indicate the utility of celastrol as a promising pharmacological agent for preventing MWCNT-induced lung tissue inflammation.

By observing the possible toxic reactions and deaths of titanium dioxide nanoparticles given by oral gavage to Sprague–Dawley (SD) rats at 2, 10, and 50 mg/kg once a day for 94 consecutive days due to accumulation, as well as the recovery of the drug for 30 days after stopping the administration of the drug, we identified the possible target organs of the toxic reactions and determined the dose of its non-toxic reaction in SD rats, in order to provide reference information for evaluating the clinical use of titanium dioxide nanoparticles and the safety of the drug. The results showed that there was no significant abnormality in body weight gain or food intake in all groups of animals during the administration period or the recovery period after drug withdrawal. Hematology, blood biochemistry, and urine indicators did not exceed the normal range, and there was no obvious toxicity. Meanwhile, when analyzed together with the histopathological results, the vital organs such as the heart, liver, lungs, kidneys, spleen, and brain did not show any obvious lesions related to the test substance. No obvious local toxicity lesions were observed in the buccal region of the mouth or the mucous membrane of the tongue at the site of drug administration. In conclusion, the non-toxic dose of titanium dioxide nanoparticles by oral gavage for 94 days in SD rats under the conditions of this test was initially determined to be 50 mg/kg/day.

Reactive oxygen species (ROS)-mediated sonodynamic therapy (SDT) holds increasing potential in treating deep-seated tumor owing to the high tissue-penetration depth. However, the inevitable accumulation of sonosensitizers in normal tissues not only make it difficult to realize the in situ SDT, but also induces sonodynamic effects in normal tissues. Herein, this work reports the passivation and selective activation strategies for the sonodynamic and near-infrared (NIR) imaging performances of an intelligent antitumor theranostic platform termed Cu-IR783 nanoparticles (NPs). Owing to the ruptured coordination bond between IR783 with Cu ions by responding to tumor microenvironment (TME), the selective activation of IR783 only occurred in tumor tissues to achieve the visualized in-situ SDT. The tumor-specific released Cu ions not only realized the cascade amplification of ROS generation through Cu+-mediated Fenton-like reaction, but also triggered cuproptosis through Cu+-induced DLAT oligomerization and mitochondrial dysfunction. More importantly, the immunosuppressive TME can be reversed by the greatly enhanced ROS levels and high-efficiency cuproptosis, ultimately inducing immunogenic cell death that promotes robust systemic immune responses for the eradication of primary tumors and suppression of distant tumors. This work provides a distinct paradigm of the integration of SDT, CDT, and cuproptosis in a controlled manner to achieve visualized in-situ antitumor therapy.

DNAJA1 is a member of type I DnaJ proteins, which is essential for spermatogenesis and male fertility. However, its expression pattern in the testes and its impact on spermatogenesis remains unclear. Our study aimed to elucidate the mechanism of action of DNAJA1. We employed DNAJA1 knockout mice in this study. Western blotting and immunofluorescence analysis were conducted to determine the protein abundance of DNAJA1 in testes at various developmental stages. Our results revealed that DNAJA1 is predominantly expressed in the testes, and its knockout leads to complete infertility in male mice. We observed that DNAJA1 protein levels increased on postnatal days 14, 21, and 28, peaking on postnatal day 35 in mice. Immunofluorescence staining indicated that DNAJA1 expression varies across different stages of the spermatogenesis cycle. Additionally, DNAJA1 was absent in epididymal sperm. In early- and mid-stage tubules, DNAJA1 protein distribution was co-localized with residual bodies in elongating spermatids. Furthermore, we found that DNAJA1 knockout significantly reduced protein polyubiquitination in the testis. Analysis of the GEO database showed that DNAJA1 levels were significantly decreased in semen samples from subjects with teratozoospermia, asthenozoospermia, and impaired spermatogenesis. Our findings suggest that DNAJA1 is an essential protein for spermatogenesis, and its deletion reduces protein polyubiquitination in the testis, ultimately resulting in infertility and spermatogenesis defects.

Endothelial barrier disruption plays a key role in the pathophysiology of heat stroke (HS). Knockout of DNAJA1 (DNAJA1‑KO) is thought to be protective against HS based on a genome‑wide CRISPR‑Cas9 screen experiment. The present study aimed to illustrate the function of DNAJA1‑KO against HS in human umbilical vein endothelial cells. DNAJA1‑KO cells were infected using a lentivirus to investigate the role of DNAJA1‑KO in HS‑induced endothelial barrier disruption. It was shown that DNAJA1‑KO could ameliorate decreased cell viability and increased cell injury, according to the results of Cell Counting Kit‑8 and lactate dehydrogenase assays. Moreover, HS‑induced endothelial cell apoptosis was inhibited by DNAJA1‑KO, as indicated by Annexin V‑FITC/PI staining and cleaved‑caspase‑3 expression using flow cytometry and western blotting, respectively. Furthermore, the endothelial barrier function, as measured by transepithelial electrical resistance and FITC‑Dextran, was sustained during HS. DNAJA1‑KO was not found to have a significant effect on the expression and distribution of cell junction proteins under normal conditions without HS. However, DNAJA1‑KO could effectively protect the HS‑induced decrease in the expression and distribution of cell junction proteins, including zonula occludens‑1, claudin‑5, junctional adhesion molecule A and occludin. A total of 4,394 proteins were identified using proteomic analysis, of which 102 differentially expressed proteins (DEPs) were activated in HS‑induced wild‑type cells and inhibited by DNAJA1‑KO. DEPs were investigated by enrichment analysis, which demonstrated significant enrichment in the 'calcium signaling pathway' and associations with vascular‑barrier regulation. Furthermore, the 'myosin light‑chain kinase (MLCK)‑MLC signaling pathway' was proven to be activated by HS and inhibited by DNAJA1‑KO, as expected. Moreover, DNAJA1‑KO mice and a HS mouse model were established to demonstrate the protective effects on endothelial barrier in vivo. In conclusion, the results of the present study suggested that DNAJA1‑KO alleviates HS‑induced endothelial barrier disruption by improving thermal tolerance and suppressing the MLCK‑MLC signaling pathway.

Embryonic-fetal developmental toxicity of compounded diphenhydramine hydrochloride (DH) and caffeine formulation has been studied by intragastric administration to pregnant SD rats. Between days 6 and 15 of pregnancy, SD rats in different groups received a low-, medium-, and high-dose of binary formulation containing DH and caffeine (DHC), a single-medication formulation containing DH, or a single-medication formulation containing caffeine. No animal deaths or clinical toxicity manifestations related to these treatments were observed in any test group. Three test groups treated with DHC formulation, as well as the DH group, showed no apparent drug influence on the body weight of pregnant rats. However, the use of caffeine alone had an impact on the body weight gain of pregnant rats (P < 0.05), which indicated maternal toxicity. The single use of caffeine alone had an impact on the food intake during pregnancy, and the food intake was recovered after medication discontinuation (P < 0.05). Except for the caffeine group, the three compounded formulation dose groups and the DH group exhibited no apparent embryonic toxicity (P > 0.05). Besides, the administration of caffeine to pregnant rats (144 mg/kg) caused fetal toxicity and influenced fetal sternal development (P < 0.05). The low, medium, and high doses of compounded DHC formulation and the single-medication DH formulation caused neither adverse reactions in pregnant rats nor impeded embryonic and fetal development. However, a single-medication formulation containing caffeine at a dose of 144 mg/kg caused toxicity to pregnant rats as well as to embryonic and fetal development.

目的 调查海军某队官兵对有毒有害化学物的认知及个人防护措施的掌握情况,为海军开展预防有毒有害化学物、增强自身防护的健康教育提供依据.方法 对海军某队218名官兵进行有毒有害化学物认知及个人防护措施掌握情况的问卷调查,记录结果并进行统计分析.结果 海军某队官兵能够较好地掌握毒理学基本常识,但是对常见的影响身体吸收有毒有害物质的外在因素和内在因素知之甚少.海军官兵对基本的个人防护措施有所了解,但缺乏对专业性防护措施的认知.结论 提高官兵对有毒有害化学物的认知并提升个人防护能力,有助于保障其身体健康,提高战斗力.

Multi-walled carbon nanotubes (MWCNTs) mainly induce oxidative stress through the overproduction of reactive oxygen species (ROS), which can lead to cytotoxicity. Celastrol, a plant-derived compound, can exert antioxidant effects by reducing ROS production. Our results indicated that exposure to MWCNTs decreased cell viability and increased ROS production. Nrf2 knockdown (kd) led to increased ROS production and enhanced MWCNT-induced cytotoxicity. Keap1-kd led to decreased ROS production and attenuated cytotoxicity. Treatment with celastrol significantly decreased ROS production and promoted Keap1 protein degradation through the lysosomal pathway, thereby enhancing the translocation of Nrf2 from the cytoplasm to the nucleus and increasing HO-1 expression. The in vivo results showed that celastrol could alleviate the inflammatory damage of lung tissues, increase the levels of the antioxidants, GSH and SOD, as well as promote the expression of the antioxidant protein, HO-1 in MWCNT-treated mice. Celastrol can alleviate MWCNT-induced oxidative stress through the Keap1/Nrf2/HO-1 signaling pathway.

A two-generation reproductive toxicity study was performed to evaluate the effects of cerium nitrate on the development of the parent, offspring, and third generation of Sprague–Dawley (SD) rats. A total of 240 SD rats (30 rats/sex/group) were randomly divided into four dosage groups according to body weight: 0 mg/kg, 30 mg/kg, 90 mg/kg, and 270 mg/kg. The rats were administered different dosages of cerium nitrate by oral gavage. There were no observed changes related to cerium nitrate in body weight, food consumption, sperm survival rate, motility, mating rate, conception rate, abortion rate, uterine plus fetal weight, uterine weight, corpus luteum number, implantation rate, live fetus number (rate), stillbirth number (rate), absorbed fetus number (rate), appearance, visceral, and skeletal in rats of each generation dosage group. In addition, the pathological findings showed no significant lesions associated with cerium nitrate toxicity in all tissues and organs, including reproductive organs. In conclusion, the present study showed that long-term oral gavage of cerium nitrate at 30 mg/kg, 90 mg/kg, and 270 mg/kg had no significant effect on reproduction and the developmental ability of their offspring in rats. The no-observed-adverse-effect level (NOAEL) of cerium nitrate in SD rats was higher than 270 mg/kg.

This study was designed to evaluate the subchronic toxicity of the compound of diphenhydramine hydrochloride (DH) and caffeine in Sprague-Dawley (SD) rats and beagle dogs. A total of 180 SD rats (15/sex/group) were randomly divided into the compound low-, medium- and high-dose groups (51, 102, 204 mg/kg), DH group (60 mg/kg), caffeine group (144 mg/kg) and the vehicle control group. Sixty beagle dogs (5/sex/group) were randomly divided into the compound low-, medium- and high-dose groups (male: 14.20, 28.30, 56.60 mg/kg, female: 5.66, 14.20, 28.30 mg/kg), DH group (male: 16.60 mg/kg, female: 8.30 mg/kg), caffeine group (male: 40.00 mg/kg, female: 20.00 mg/kg) and the vehicle control group. Rats and dogs were given continuous oral administration for 28 days following a 28-day recovery period. The adverse effects of the compound on rats and beagle dogs mainly included anorexia and liver function impairment. Most adverse effects induced by administration were reversible. Under the experimental conditions, the no-observed-adverse-effect level (NOAEL) of the compound of DH and caffeine was 51 mg/kg/day for SD rats and 28.30 mg/kg/day (male) and 5.66 mg/kg/day (female) for beagle dogs.

Although low‐cost nanozymes with excellent stability have demonstrated the potential to be highly beneficial for nanocatalytic therapy (NCT), their unsatisfactory catalytic activity accompanied by intricate tumor microenvironment (TME) significantly hinders the therapeutic effect of NCT. Herein, for the first time, a heterojunction (HJ)‐fabricated sonoresponsive and NIR‐II‐photoresponsive nanozyme is reported by assembling carbon dots (CDs) onto TiCN nanosheets. The narrow bandgap and mixed valences of Ti 3+ and Ti 4+ endow TiCN with the capability to generate reactive oxygen species (ROS) when exposed to ultrasound (US), as well as the dual enzyme‐like activities of peroxidase and glutathione peroxidase. Moreover, the catalytic activities and sonodynamic properties of the TiCN nanosheets are boosted by the formation of HJs owing to the increased speed of carrier transfer and the enhanced electron–hole separation. More importantly, the introduction of CDs with excellent NIR‐II photothermal properties could achieve mild hyperthermia (43 °C) and thereby further improve the NCT and sonodynamic therapy (SDT) performances of CD/TiCN. The synergetic therapeutic efficacy of CD/TiCN through mild hyperthermia‐amplified NCT and SDT could realize “three‐in‐one” multimodal oncotherapy to completely eliminate tumors without recurrence. This study opens a new avenue for exploring sonoresponsive and NIR‐II‐photoresponsive nanozymes for efficient tumor therapy based on semiconductor HJs.

Hepatocyte transplantation can be an effective treatment for patients with certain liver-based metabolic disorders and liver injuries. Hepatocytes are usually infused into the portal vein, from which hepatocytes migrate into the liver and integrate into the liver parenchyma. However, early cell loss and poor liver engraftment represent major hurdles to sustaining the recovery of diseased livers after transplantation. In the present study, we found that ROCK (Rho-associated kinase) inhibitors significantly enhanced in vivo hepatocyte engraftment. Mecha-nistic studies suggested that the isolation of hepatocytes caused substantial degradation of cell membrane proteins, including the complement inhibitor CD59, probably due to shear stress-induced endocytosis. ROCK inhibition by ripasudil, a clinically used ROCK inhibitor, can protect transplanted hepatocytes by retaining cell membrane CD59 and blocking the formation of the membrane attack complex. Knockdown of CD59 in hepatocytes eliminates ROCK inhibition-enhanced hepatocyte engraftment. Ripasudil can accelerate liver repopu-lation of fumarylacetoacetate hydrolase-deficient mice. Our work reveals a mechanism underlying hepatocyte loss after transplantation and provides immediate strategies to enhance hepatocyte engraftment by inhibiting ROCK.

PBL教学模式的应用为课程教育带去了更多契机，有助于打破传统教育理念的束缚，促进学生发展。教师通过教学教 案的提前预设，将PBL教学模式和教学内容相结合，能够更好地提高学生应用、分析问题的能力，从而为学生今后进 入社会就业打好基础。本文对PBL教学模式进行了多角度的研究，旨在为教育工作者提供参考意见。

In this study, the toxicity of ferric oxide nanoparticles (Fe2O3 NPs) administered through gavage to Sprague Dawley (SD) rats for 94 d, consecutively and the recovery after Fe2O3 NPs withdrawal for 30 d were evaluated. The vehicle control group, low-, medium-, and high-dose groups were administered with the vehicle (0.5% sodium carboxymethyl cellulose [CMC-Na]), 125, 250, and 500 mg/kg of Fe2O3 NPs, respectively, administered every morning for 94 d. There was no significant difference in the body weight, food intake, hematological, blood biochemical, and urine indices of SD rats in each administration group and the control group (P > 0.05). There was no significant difference in organ weight, organ indices, and the coefficient of the visceral brain between the SD rats in the different dosage groups and the SD rats in the vehicle control group (P > 0.05). Histopathological observations showed that there was no correlation between the pathological lesions of the organs observed in this study and the dose of Fe2O3 NPs (P > 0.05). The no-observed-adverse-effect level (NOAEL) dose of Fe2O3 NPs was initially determined to be 500 mg/kg administered to SD rats through oral gavage for 94 d, consecutively, followed by recovery after Fe2O3 NPs withdrawal for 30 d.