Disruption of Zfh3 abolishes mulberry-specific monophagy in silkworm larvae.

Feeding behavior is critical for insect survival and fitness. Most researchers have explored the molecular basis of feeding behaviors by identifying and elucidating the function of olfactory receptors (ORs) and gustatory receptors (GRs). Other types of genes, such as transcription factors, have rarely been investigated, and little is known about their potential roles. The silkworm (Bombyx mori) is a well-studied monophagic insect which primarily feeds on mulberry leaves, but the genetic basis of its monophagy is still not understood. In this report, we focused on a transcription factor encoded by the Zfh3 gene, which is highly expressed in the silkworm central and peripheral nervous systems, including brain, antenna, and maxilla. To investigate its function, Zfh3 was abrogated using clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated protein 9 (Cas9) mutagenesis. Since Zfh3 knockout homozygotes are not viable, we studied feeding behavior in heterozygotes, and found that disruption of Zfh3 affects both gustation and olfaction. Mutant larvae lose preference for mulberry leaves, acquire the ability to consume an expanded range of diets, and exhibit improved adaptation to the M0 artificial diet, which contains no mulberry leaves. These results provide the first demonstration that a transcription factor modulates feeding behaviors in an insect.

Double-layer hollow mesoporous silica nanoparticles for ultrasound-guided photodynamic treatment.

Cervical carcinoma persists as a major global public health burden. While conventional therapeutic modalities inevitably cause ablation of adjacent non-tumorous tissues, photodynamic therapy (PDT) offers a targeted cytotoxic strategy through a photosensitizing agent (PS). However, the hydrophobicity and lack of selective accumulation of promising PS compounds such as zinc(II) phthalocyanine (ZnPc) impedes their clinical translation as standalone agents. The present study sought to incorporate ZnPc within double-layer hollow mesoporous silica nanoparticles (DHMSN) as nanocarriers to enhance aqueous dispersibility and tumor specificity. Owing to their compartmentalized design, the hollow mesoporous silica nanoparticles (HMSN) demonstrated enhanced ultrasonic imaging contrast. Combined with the vaporization of the perfluorocarbon perfluoropentane (PFP), the HMSN-encapsulated ZnPc enabled real-time ultrasound monitoring of PDT treatment.In vivo, the innate thermal energy induced vaporization of the DHMSN-carried PFP to significantly amplify ultrasound signals from the tumor site. Results demonstrated biocompatibility, efficient PFP microbubble generation, and robust photocatalytic activity. Collectively, this investigation establishes ultrasound-guided PDT utilizing multi-layer HMSN as a targeted therapeutic strategy for cervical malignancies with mitigated toxicity.

Super silkworm cocoons constructed by multi-silkworm larvae: Promising composites with dense structures and excellent mechanical properties.

A normal silkworm cocoon (NSC) with a unique nonwoven structure is usually spun by a single silkworm larva. Notably, there is a special Bombyx mori genetic resource that many (three or more) mature larvae tend to collectively spin into one cocoon, which was named "multi-silkworm cocoon" ("MSC"). However, the MSCs display loose structure and poor mechanical properties which limits their further application. In this study, a series of hybrid silkworm cocoons (HMSCs) are obtained by hybridizing "MSC" with a selected commercial silkworm strain successfully. The morphology, microstructures, and mechanical properties of cocoons constructed by one to three silkworm larvae were characterized and compared. The results indicated that about 48.3 % of silkworm larvae could create double and triple cocoons in the F1 generation of the silkworm hybrid, displaying robust fiber networks and dense structures. The mechanical characteristics of the HMSCs, including the tensile, peeling, compression, and needle penetration resistance properties, exceeded those of MSCs, showing significant application potential for high-performance bio-composites. This study provides a practical approach for obtaining silkworm cocoons with controllable structures and mechanical properties to develop and fabricate natural composite and biomimetic materials.

Enhanced anti-glioma efficacy of biodegradable periodic mesoporous organosilica nanoparticles through target delivery of chemotherapeutics.

Glioma is the most common malignant tumor of the brain and enhancing the efficacy of chemotherapy in glioma is critical for improving patients' prognosis. In this study, a glioma-targeting drug delivery system is constructed using biodegradable periodic mesoporous organosilica nanoparticles (PMO) that are modified with lactoferrin (Lf) ligands. The obtained PMO is doped with thioether groups and can be degraded in the high concentration of glutathione in tumor cells. The surface area and pore volume of PMO are 772 cm2/g and 0.98 cm3/g, respectively and the loading capacity of doxorubicin (Dox) is as high as 20%. The results of the confocal laser scanning microscope show that the uptake of PMO-Lf@Dox by C6 cells is higher than PMO@Dox. The quantitative analysis of the flow cytometer further demonstrates that more PMO-Lf@Dox enter C6 cells, indicating that the modification of lactoferrin can significantly increase the uptake of C6 cells. Finally, the therapeutic efficacy results show that Lf-modified PMO enhances the inhibitory effect of Dox on C6 cells when incubated for 24 h and 72 h. In summary, this lactoferrin receptor-mediated PMO drug carrier with biodegradability in glutathione in tumor cells can be used to enhance drug delivery into glioma without long-term accumulation in vivo. In this study, a glioma-targeting drug delivery system is constructed using periodic mesoporous organosilica nanoparticles (PMO) that modified with lactoferrin (Lf) ligands. This lactoferrin receptor-mediated PMO drug carrier can be used to enhance drug delivery into brain glioma.

Interpretable XGBoost-SHAP Model Predicts Nanoparticles Delivery Efficiency Based on Tumor Genomic Mutations and Nanoparticle Properties.

Understanding the complex interaction between nanoparticles (NPs) and tumors in vivo and how it dominates the delivery efficiency of NPs is critical for the translation of nanomedicine. Herein, we proposed an interpretable XGBoost-SHAP model by integrating the information on NPs physicochemical properties and tumor genomic profile to predict the delivery efficiency. The correlation coefficients were 0.66, 0.75, and 0.54 for the prediction of maximum delivery efficiency, delivery efficiency at 24 and 168 h postinjection for test sets. The analysis of the feature importance revealed that the tumor genomic mutations and their interaction with NPs properties played important roles in the delivery of NPs. The biological pathways of the NP-delivery-related genes were further explored through gene ontology enrichment analysis. Our work provides a pipeline to predict and explain the delivery efficiency of NPs to heterogeneous tumors and highlights the power of simultaneously using omics data and interpretable machine learning algorithms for discovering interactions between NPs and individual tumors, which is important for the development of personalized precision nanomedicine.

Establishment of a nomogram for predicting functional constipation among children in China: Using the Rome IV criteria.

BACKGROUND AND STUDY AIMS: Childhood functional constipation (FC) is gradually becoming an emerging public health problem. This study aimed to develop a personalized nomogram for the prediction of incident FC among Chinese children, and the diagnosis of FC was based on the Rome IV criteria. PATIENTS AND METHODS: This cross-sectional study was conducted from Nov. 2020 to Jan. 2021 among children residing in Anhui province, China. An electronic questionnaire regarding the general demographic and clinical characteristics of all children was completed by their primary caregivers. The multivariate logistic regression analysis was applied to identify risk factors for FC. Moreover, a nomogram was constructed for FC based on the risk factors identified from the multivariate analysis. RESULTS: In this study, a total of 901 electronic questionnaires were collected, of which 832 (92.3%) questionnaires were properly completed and included in the final analysis. The prevalence of FC among Chinese children was 11.3% based on the Rome IV criteria. After controlling for potential confounding factors, the multivariate logistic regression analysis showed that inadequate sleep, picky eating, and positive family history of FC were identified as key risk factors of FC. The area under the receiver operating characteristic curve of the nomogram was 0.694 (95 %CI: 0.6412-0.7459). Further, a calibration curve drawn illustrated that the predicted probabilities reasonably approximately the actual prevalence of FC in this population. CONCLUSION: Inadequate sleep, picky eating, and positive family history of FC were identified as risk factors of FC. An easy-to-use nomogram was constructed based on these three significant factors. Besides, this nomogram was validated to have acceptable discrimination and calibration capabilities. Hence, this nomogram may enable clinical professionals to predict the risk of FC among Chinese children and further provide optimized disease prevention and intervention for this population.

Theranostic mesoporous platinum nanoplatform delivers halofuginone to remodel extracellular matrix of breast cancer without systematic toxicity.

The enriched collagens in the extracellular matrix (ECM) of breast cancer substantially impede drug delivery. Halofuginone (HF), a potent antifibrotic agent, was effective to deplete the collagens and remodel the ECM by inhibiting the TGFß pathway. However, the application of HF was hindered by its strong liver toxicity. Herein, mesoporous platinum (mPt) nanoparticles were constructed to load HF as theranostic nanoplatforms. mPt had a uniform spherical structure with a diameter of 79.83 ± 6.97 nm and an average pore diameter of 20 nm and exhibited good photothermal conversion efficiency of 62.4%. The obtained HF-loaded nanoplatform (PEG@mPt-HF) showed enhanced cytotoxicity through the combination of photothermal therapy and the anti-TGFß effect induced by HF. The animal imaging and histochemical assays confirmed the PEG@mPt-HF could efficiently deliver HF to tumors (monitored by CT) and remodel the ECM by TGFß pathway inhibition, which resulted in increased anti-cancer efficacy. Importantly, the liver toxicity observed in HF-treated mice was negligible in those treated by PEG@mPt-HF. Overall, this study designed a theranostic nanoplatform to remodel the ECM with remarkably reduced systematic toxicity and enhance the therapeutic efficacy through combination treatment.

The value of DCE- MRI of the breast as a diagnostic tool in assessing amorphous calcifications in screening mammography.

Purpose: To evaluate the diagnostic performance of dynamic contrast-enhanced magnetic resonance imaging in differentiating benign and malignant amorphous calcifications. Methods: This study included 193 female patients with 197 suspicious amorphous calcifications detected on screening mammography. The patients' demographics, clinical follow-up, imaging, and pathology outcomes were reviewed, and sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of DCE-MRI were calculated. Results: Of 197 lesions (193 patients) included in the study, 50 (25.4%) were histologically proved to be malignant. DCE-MRI based on breast imaging report and diagnosis system (BI-RADS) had a sensitivity of 94.4%, specificity of 85.7%, PPV of 69.1%, and NPV of 97.7% for the detection of malignant amorphous calcifications. Notably, diagnosis solely based on the presence or absence of DCE-MRI enhancement showed the same sensitivity but significantly decreased specificity (44.8%, p < 0.001) and PPV (44.8%, p < 0.001). In patients with a minimal or mild degree of background parenchymal enhancement (BPE), the sensitivity, specificity, PPV, and NPV increased to 100%, 90.6%, 78.6%, and 100%, respectively. However, in patients with a moderate degree of BPE, MRI resulted in three false negatives of ductal carcinoma in situ (DCIS). Overall, the addition of DCE-MRI detected all invasive lesions and could decrease unnecessary biopsy by 65.5%. Conclusion: DCE-MRI based on BI-RADS has the potential to improve the diagnosis of suspicious amorphous calcifications and avoid unnecessary biopsy, especially for those with low-degree BPE.

Tumor-targeting semiconducting polymer nanoparticles: efficient adjuvant photothermal therapy using ultra-low laser power inhibits recurrences after breast-conserving surgery.

The need for adjuvant therapy to inhibit local recurrence after breast-conserving surgery with minimal side effects is great. Adjuvant photothermal therapy (aPTT) has the potential to replace radiotherapy and eliminates its inherent damage to healthy tissues. Herein, we functionalized semiconducting polymer nanoparticles (SPNs) with cRGD-peptide and silicon 2,3-naphthalocyanine bis(trihexylsilyloxide) (NIR775) to target breast cancer and perform aPTT under an ultra-low laser power (0.2 W cm-2) after breast-conserving surgery (BCS). The synthesized RGD-SPNNIR775 showed an excellent photothermal conversion efficiency and biocompatibility and was demonstrated to accumulate in tumors specifically. The BCS could be performed with confidence under the guidance of preoperative and postoperative fluorescence imaging. Notably, the aPTT completely inhibited the local recurrence after the BCS without compromising the cosmetic effect of the BCS. These results indicate the prospect of RGD-SPNNIR775 as a theranostic nanoplatform for efficient aPTT using an ultra-low laser power to control recurrence after BCS.

Cross-cultural adaptation and validation of the geriatric 8 screening tool in Chinese hospitalized older adults with cancer.

OBJECTIVES: To translate, cross-culturally adapt, and validate the Geriatric 8 (G8) questionnaire in Chinese hospitalized older adults with cancer. METHODS: The Chinese version of the G8 (C-G8) was produced following Brislin's guidelines. The psychometric properties of the C-G8 were evaluated among 296 eligible patients. RESULTS: The content validity index of the C-G8 was 0.8∼1 at the item level and 0.975 at the scale level. The C-G8 identified more frail individuals among these older (>75 years) participants compared to their younger (65∼75 years) counterparts (frailty prevalence: 87.1% vs. 70.9%, P=0.010). The convergent validity of the C-G8 was tested by correlating it with the FRAIL scale (r=-0.592, P<0.001). The C-G8 had a lower internal consistency (Cronbach's α coefficient=0.501) but higher test-retest reliability and inter-rater reliability (intraclass correlation coefficient=0.913 and 0.993, respectively, P<0.001). CONCLUSIONS: The C-G8 questionnaire presented acceptable validity and reliability and could be used in Chinese hospitalized older adults with cancer.

High-resolution silkworm pan-genome provides genetic insights into artificial selection and ecological adaptation.

The silkworm Bombyx mori is an important economic insect for producing silk, the "queen of fabrics". The currently available genomes limit the understanding of its genetic diversity and the discovery of valuable alleles for breeding. Here, we deeply re-sequence 1,078 silkworms and assemble long-read genomes for 545 representatives. We construct a high-resolution pan-genome dataset representing almost the entire genomic content in the silkworm. We find that the silkworm population harbors a high density of genomic variants and identify 7308 new genes, 4260 (22%) core genes, and 3,432,266 non-redundant structure variations (SVs). We reveal hundreds of genes and SVs that may contribute to the artificial selection (domestication and breeding) of silkworm. Further, we focus on four genes responsible, respectively, for two economic (silk yield and silk fineness) and two ecologically adaptive traits (egg diapause and aposematic coloration). Taken together, our population-scale genomic resources will promote functional genomics studies and breeding improvement for silkworm.

Simultaneous glutamine metabolism and PD-L1 inhibition to enhance suppression of triple-negative breast cancer.

Blockade of programmed cell death 1 ligand (PD-L1) has been used to treat triple-negative breast cancer (TNBC), and various strategies are under investigation to improve the treatment response rate. Inhibition of glutamine metabolism can reduce the massive consumption of glutamine by tumor cells and meet the demand for glutamine by lymphocytes in tumors, thereby improving the anti-tumor effect on the PD-L1 blockade therapy. Here, molybdenum disulfide (MoS2) was employed to simultaneously deliver anti-PDL1 antibody (aPDL1) and V9302 to boost the anti-tumor immune response in TNBC cells. The characterization results show that MoS2 has a dispersed lamellar structure with a size of about 181 nm and a size of 232 nm after poly (L-lysine) (PLL) modification, with high stability and biocompatibility. The loading capacity of aPDL1 and V9302 are 3.84% and 24.76%, respectively. V9302 loaded MoS2 (MoS2-V9302) can effectively kill 4T1 cells and significantly reduce glutamine uptake of tumor cells. It slightly increases CD8+ cells in the tumor and promotes CD8+ cells from the tumor edge into the tumor core. In vivo studies demonstrate that the combination of aPDL1 and V9302 (MoS2-aPDL1-V9302) can strongly inhibit the growth of TNBC 4T1 tumors. Interestingly, after the treatment of MoS2-aPDL1-V9302, glutamine levels in tumor interstitial fluid increased. Subsequently, subtypes of cytotoxic T cells (CD8+) in the tumors were analyzed according to two markers of T cell activation, CD69, and CD25, and the results reveal a marked increase in the proportion of activated T cells. The levels of cytokines in the corresponding tumor interstitial fluid are also significantly increased. Additionally, during the treatment, the body weights of the mice remain stable, the main indicators of liver and kidney function in the blood do not increase significantly, and there are no obvious lesions in the main organs, indicating low systemic toxicity. In conclusion, our study provides new insights into glutamine metabolism in the tumor microenvironment affects immune checkpoint blockade therapy in TNBC, and highlights the potential clinical implications of combining glutamine metabolism inhibition with immune checkpoint blockade in the treatment of TNBC.

Periodic mesoporous organosilica coupled with chlorin e6 and catalase for enhanced photodynamic therapy to treat triple-negative breast cancer.

Photodynamic therapy (PDT) has become a promising treatment option for highly aggressive triple-negative breast cancer (TNBC); however, hypoxia limits the efficacy of PDT and promotes tumour aggression. In this work, we first constructed a multifunctional yolk-shell structured nanoplatform consisting of periodic mesoporous organosilica (PMO) coupled with chlorin e6 (Ce6) and catalase (Catalase) (Yolk-Shell PMO-Ce6@Catalase) for enhanced PDT against TNBC. This nanoplatform has an organic-inorganic hybrid skeleton structure, a uniform size and good stability and biocompatibility. In vitro experiments showed that the nanoplatform has a good ability to generate singlet oxygen. Catalase can convert H2O2 into O2, increasing the concentration of oxygen around the cells and overcoming the problem of hypoxia in the tumour, which enhances the effects of PDT. The in vivo experimental results showed that PDT with the Yolk-Shell PMO-Ce6@Catalase nanoplatform, compared with free Ce6 and Yolk-Shell PMO-Ce6 PDT, can significantly inhibit tumour growth, revealing the most extensive cellular apoptosis and necrosis in the tumour area in this treatment group. Additionally, the histopathological results showed that PDT did not cause significant side effects to the major organs. Therefore, we believe that this Yolk-Shell PMO-Ce6@Catalase nanoplatform has excellent clinical potential for PDT against TNBC.

Hybridized double-shell periodic mesoporous organosilica nanotheranostics for ultrasound imaging guided photothermal therapy.

Hybridized periodic mesoporous organosilica (PMO) nanoparticles are expected to provide a multifunctional theranostic platform for precision medicine by combining the advantages of different organic and inorganic components. In this work, double-shell-structured PMO nanotheranostics composed of ethane- and thioether-bridged organosilica shells were synthesized. Gold colloids were generated in situ by the thioether groups on the inner shell. The obtained double-shell PMO@Au (DSPA) has uniform size, large surface areas, ordered mesochannels and photothermal conversion capability. After being encapsulated with perfluorohexacene (PFH), DSPA-PFH produced a strong ultrasound signal upon laser irradiation due to the phase transit of PFH during hyperthermia. DSPA-PFH showed enhanced photothermal therapeutic efficacy, great ultrasound contrast, and minimal toxicity both in vitro and in vivo. These results demonstrated the distribution of different organosilica could be delicately adjusted in hybridized PMO nanoparticles. Furthermore, it showed the potential of using hybridized PMO nanoparticles as a theranostic platform for biomedical applications by combining unique characteristics of different organosilica through rational design.

Disrupting stromal barriers to enhance photothermal-chemo therapy using a halofuginone-loaded Janus mesoporous nanoplatform.

Dense tumor stroma is the physiological barrier in drug delivery that prevents anticancer drugs from entering the tumor, thereby seriously limiting the drugs' therapeutic effect. In this study, a Janus nanoplatform consisting of periodic mesoporous organosilica-coated platinum nanoplatforms (JPMO-Pt) and anti-stroma drug halofuginone (HF) (denoted as JPMO-Pt-HF), was developed to deplete the tumor stroma and synergistically treat breast cancer in BALB/c mice. The prepared JPMO-Pt had a uniform size of 245 nm, a good dispersion, an excellent in vitro and in vivo biocompatibility, and a high loading capacity for HF (up to 50 µg/mg). The antitumor experiments showed that the survival rate of 4 T1 cells exhibited an obvious downward trend when the cells were incubated with the JPMO-Pt-HF and irradiated with 808 nm laser. Moreover, the cell survival rate was only about 10% at 48 h when the HF concentration was 2.0 µg/mL. Notably, JPMO-Pt-HF under irradiation had an excellent synergistic therapeutic effect on tumor cells. In vivo antitumor experiment further showed that the JPMO-Pt-HF, in combination with laser irradiation, could minimize tumor growth, showing significantly better effects than those observed for the case of monotherapy involving photothermal therapy (PTT) (152 vs. 670 mm3, p < 0.0001) and HF (152 vs. 419 mm3, p = 0.0208). In addition, immunohistochemistry of tumor tissues indicated that JPMO-Pt-HF obviously reduced the relative collagen and α-smooth muscle actin (α-SMA) area fraction. Taken together, this research designs a new platform that not only possesses the ability to degrade the tumor matrix but also combines PTT and chemotherapeutic effects, and holds promise for effective tumor treatment.

Chlorin e6-Biotin Conjugates for Tumor-Targeting Photodynamic Therapy.

To improve the tumor-targeting efficacy of photodynamic therapy, biotin was conjugated with chlorin e6 to develop a new tumor-targeting photosensitizer, Ce6-biotin. The Ce6-biotin had good water solubility and low aggregation. The singlet-oxygen generation rate of Ce6-biotin was slightly increased compared to Ce6. Flow cytometry and confocal laser scanning microscopy results confirmed Ce6-biotin had higher binding affinity toward biotin-receptor-positive HeLa human cervical carcinoma cells than its precursor, Ce6. Due to the BR-targeting ability of Ce6-biotin, it exhibited stronger cytotoxicity to HeLa cells upon laser irradiation. The IC50 against HeLa cells of Ce6-biotin and Ce6 were 1.28 µM and 2.31 µM, respectively. Furthermore, both Ce6-biotin and Ce6 showed minimal dark toxicity. The selectively enhanced therapeutic efficacy and low dark toxicity suggest that Ce6-biotin is a promising PS for BR-positive-tumor-targeting photodynamic therapy.

Dye-loaded mesoporous polydopamine nanoparticles for multimodal tumor theranostics with enhanced immunogenic cell death.

BACKGROUND: Tumor phototherapy especially photodynamic therapy (PDT) or photothermal therapy (PTT), has been considered as an attractive strategy to elicit significant immunogenic cell death (ICD) at an optimal tumor retention of PDT/PTT agents. Heptamethine cyanine dye (IR-780), a promising PDT/PTT agent, which can be used for near-infrared (NIR) fluorescence/photoacoustic (PA) imaging guided tumor phototherapy, however, the strong hydrophobicity, short circulation time, and potential toxicity in vivo hinder its biomedical applications. To address this challenge, we developed mesoporous polydopamine nanoparticles (MPDA) with excellent biocompatibility, PTT efficacy, and PA imaging ability, facilitating an efficient loading and protection of hydrophobic IR-780. RESULTS: The IR-780 loaded MPDA (IR-780@MPDA) exhibited high loading capacity of IR-780 (49.7 wt%), good physiological solubility and stability, and reduced toxicity. In vivo NIR fluorescence and PA imaging revealed high tumor accumulation of IR-780@MPDA. Furthermore, the combined PDT/PTT of IR-780@MPDA could induce ICD, triggered immunotherapeutic response to breast tumor by the activation of cytotoxic T cells, resulting in significant suppression of tumor growth in vivo. CONCLUSION: This study demonstrated that the as-developed compact and biocompatible platform could induce combined PDT/PTT and accelerate immune activation via excellent tumor accumulation ability, offering multimodal tumor theranostics with negligible systemic toxicity.

GANDA: A deep generative adversarial network conditionally generates intratumoral nanoparticles distribution pixels-to-pixels.

Intratumoral nanoparticles (NPs) distribution is critical for the success of nanomedicine in imaging and treatment, but computational models to describe the NPs distribution remain unavailable due to the complex tumor-nano interactions. Here, we develop a Generative Adversarial Network for Distribution Analysis (GANDA) to describe and conditionally generates the intratumoral quantum dots (QDs) distribution after i.v. injection. This deep generative model is trained automatically by 27,775 patches of tumor vessels and cell nuclei decomposed from whole-slide images of 4 T1 breast cancer sections. The GANDA model can conditionally generate images of intratumoral QDs distribution under the constraint of given tumor vessels and cell nuclei channels with the same spatial resolution (pixels-to-pixels), minimal loss (mean squared error, MSE = 1.871) and excellent reliability (intraclass correlation, ICC = 0.94). Quantitative analysis of QDs extravasation distance (ICC = 0.95) and subarea distribution (ICC = 0.99) is allowed on the generated images without knowing the real QDs distribution. We believe this deep generative model may provide opportunities to investigate how influencing factors affect NPs distribution in individual tumors and guide nanomedicine optimization for molecular imaging and personalized treatment.

Self-assembled small messenger RNA nanospheres for efficient therapeutic apoptin expression and synergistic Gene-Chemotherapy of breast cancer.

In this work, small self-assembled messenger RNA nanospheres (mRNA-NSs) were successfully prepared by rolling circle transcription on a constructed apoptin plasmid. The self-assembled mRNA-NSs have a uniform diameter of approximately 65 nm, good dispersity in solution, and efficient therapeutic apoptin expression ability. In addition, the mRNA-NSs have a high loading capacity of 8.2% for the antitumor drug doxorubicin (Dox), which can effectively deliver the loaded Dox into 4 T1 cells. Cellular experiments show that Dox-loaded self-assembled messenger RNA nanospheres (mRNA-NSs@Dox) can reduce the viability of 4 T1 breast cancer cells by significantly upregulating Bax protein, thereby inducing the activation of Caspase 3 in 4 T1 cells. In vivo experiments show that mRNA-NSs@Dox can effectively increase the necrosis of tumor tissue, reduce the expression of Ki67, and exhibit a synergistic gene-chemotherapy effect in breast cancer-bearing mice. Taken together, this study successfully prepared self-assembled apoptin messenger RNA nanospheres (mRNA-NSs), which can improve the expression of the therapeutic protein apoptin and exhibit excellent synergistic antitumor effects after loading Dox, providing new ideas for the gene treatment and chemotherapy of breast cancer.