1.Development trajectories of clustered health risk behaviors and their association with self esteem and loneliness among junior high school students
XU Tao, LIU Lü ; hao, CHEN Jiajia, WANG Yongsheng, ZHANG Tiancheng, YANG Xuejing
Chinese Journal of School Health 2026;47(2):189-193
Objective:
To analyze the developmental trajectories of clustered health risk behaviors and their association with self-esteem and lonelinesss among junior high school students, so as to provide a reference for formulating comprehensive prevention and control measures of health risk behaviors among adolescents.
Methods:
In October 2023, 1 165 first year junior high school students from two schools of Jishou City in Hunan Province were selected by convenient sampling method for three follow up surveys (T1:October 2023; T2:April 2024; T3:October 2024). The Adolescent Health Risk Behavior Questionnaire, Rosenberg Self esteem Scale and Loneliness Scale were used to assess health risk behaviors, self esteem and loneliness, respectively. Latent growth curve modeling and latent growth mixture modeling were applied to analyze the developmental trajectories of clustered health risk behaviors among junior high school students. Logistic regression was used to analyze the association of the developmental trajectories of clustered health risk behaviors with self esteem and loneliness among junior high school students.
Results:
The overall developmental trajectories among junior high school students showed a declining trend (intercept=0.15, slope=-1.65, both P <0.05), with three heterogeneous categories:low risk improvement group ( n =862, 74.0%), moderate risk stable group ( n =260, 22.3%), and high risk deterioration group ( n =43, 3.7%). After adjusting the status of the left behind individuals,using the low risk improvement group as the reference category in multinomial Logistic regression analysis, results indicated that higher loneliness scores among junior high school students increased the risks of belonging to the moderate risk stable group ( OR=1.02, 95%CI =1.00- 1.04 ) and the high risk deterioration group ( OR=1.04, 95%CI =1.00-1.08), while higher self esteem scores reduced the risks of belonging to the moderate risk stable group ( OR=0.93, 95%CI =0.91-0.96) and the high risk deterioration group ( OR=0.88, 95%CI =0.83-0.94) (all P <0.05).
Conclusions
The overall trend of clustered health risk behaviors among junior high school students gradually improves, and the self esteem and loneliness are significant correlative factors. Targeted intervention measures should be developed for the junior high school students, with a focus on enhancing their self esteem and alleviating loneliness.
2.Association of outdoor activity time and sleep duration with screening myopia in primary school students
LIU Xiaoling, LI Tingting, CAO Caiyun, YANG Feng, TAO Shuman, WU Xiaoyan, XU Shaojun, TAO Fangbiao
Chinese Journal of School Health 2026;47(2):279-282
Objective:
To explore the association of outdoor activity time and sleep duration with screening myopia in primary school students, so as to provide strategies for myopia prevention.
Methods:
Through a convenience sampling method, a survey was conducted among 4 248 primary school students aged 7-13 years from three primary schools in Xihu District, Nanchang City, Jiangxi Province from May to July, 2023. The average daily outdoor activity time and sleep duration on both weekdays and weekends were investigated in primary school students by using a self designed questionnaire. Uncorrected visual acuity tests and non cycloplegic autorefraction were measured by professional optometrists. Inter group comparisons were conducted using the Chi square test. Logistic regression model was used to analyze the association of outdoor activity time and sleep duration with screening myopia.
Results:
The detection rate of screening myopia in primary school students was 33.6%, with the rate in boys (32.0%) lower than that in girls (35.3%), and the difference was statistically significant ( χ 2=5.11, P =0.02). The analysis results of Logistic regression showed that after adjusting for factors such as gender, grade and parental education level, both average daily outdoor activity time <2 h on both weekdays and weekends ( OR =1.27, 95% CI =1.11-1.46) and sleep duration <10 h ( OR =1.17, 95% CI =1.01- 1.35 ), as well as their combined effect ( OR =1.57, 95% CI =1.25-1.98), were associated with an increased risk of screening myopia in primary school students(all P <0.05). Subgroup analysis results indicated that compared to boys ( OR =1.46, 95% CI = 1.07 -1.99), girls( OR =1.73, 95% CI =1.22-2.44) with insufficient outdoor activity time and sleep duration had a higher risk of screening myopia(both P <0.05).
Conclusions
There is a negative correlation of outdoor activity time and sleep duration with screening myopia in primary school students. Outdoor activity time and extending sleep duration should be increased to reduce the risk of myopia in primary school students.
3.Evaluation of the application effectiveness and optimization strategies of confidential unit exclusion in Zhengzhou
Dan LIU ; Hongwei MA ; Tao WEN ; Yonglei LYU ; Mengru JI ; Ge SONG ; Huanyu LIU ; Mengdi FAN
Chinese Journal of Blood Transfusion 2026;39(3):379-383
Objective: To evaluate the practical effectiveness of confidential unit exclusion (CUE) in ensuring blood safety in Zhengzhou, analyze its application characteristics and existing problems, and provide a basis for optimizing blood safety management strategies. Methods: A retrospective analysis was conducted on CUE data handled by Henan Red Cross Blood Center from January 2019 to December 2024. Parameters such as the number of cases, demographic characteristics, reasons for exclusion, and time of report were statistically analyzed and compared with those of non-CUE. Results: From 2019 to 2024, the CUE reporting rate in Zhengzhou was 0.002 6% (40/1 547 666). CUE donors were predominantly male (65.00%, 26/40), aged 18-34 years (47.50%, 19/40), had college degree orabove (50.00%, 20/40), and were employees of enterprises or public institutions (32.50%, 13/40). Among the 40 CUE blood units, only one was reactive for anti-TP, while all others were qualified. The main reasons for CUE were recent vaccination (32.50%, 13/40), medical conditions unsuitable for donation (27.50%, 11/40), and high-risk sexual behavior (17.50%, 7/40). A total of 70.00% of reports occurred within 24 hours after donation, during which none of the corresponding blood units had been released; all units reported after more than 7 days had already been issued for clinical use, with no adverse transfusion reactions reported upon follow-up. Conclusion: The confidential unit exclusion program has played an active role in establishing a supplementary information feedback channel for blood donors. The procedure can be optimized by strengthening interactive communication and confirmation before donation, improving the accuracy of donors' self-assessment, and expanding convenient and rapid information-based reporting channels.
4.Effect of mesaconate combined with LR on prolonging the golden treatment time window and its resuscitation efficacy for hemorrhagic shock rats under high-altitude conditions
Yuanqun ZHOU ; Xinming XIANG ; Xingnan OUYANG ; Jie ZHANG ; Qinghui LI ; Liangming LIU ; Tao LI
China Pharmacy 2026;37(6):720-726
OBJECTIVE To observe the effects of endogenous metabolite mesaconate combined with Sodium lactate Ringer’s injection (LR) on prolonging the golden treatment time window and its resuscitation efficacy in rats with hemorrhagic shock under high-altitude conditions. METHODS Rats were divided into the shock group, LR group, and 5, 20, 50 mg/kg mesaconate+LR groups, with 20 rats in each group, to investigate the effect of additional use of mesaconate on the golden treatment time window. After establishing a model of uncontrolled hemorrhagic shock under high-altitude conditions in all groups by housing in a hypobaric hypoxia chamber combined with splenic artery transection, rats in the shock group received no resuscitation, while rats in the LR group and mesaconate+LR groups underwent low-pressure resuscitation with LR or mesaconate combined with LR. Blood pressure control, fluid infusion volume, blood loss rate and survival status were observed in each group. Rats were further divided into the normal group, shock group and mesaconate (50 mg/kg)+LR group, with 10 or 20 rats in each group, to evaluate the resuscitation effects after extending the golden treatment time window by additionally using mesaconate. Except for the normal group, the other groups underwent the same procedure to establish an uncontrolled hemorrhagic shock model under high-altitude conditions. Rats in the shock group received no resuscitation. In the mesaconate+LR group, after 3 h of low-pressure resuscitation, bleeding control was performed by ligation of the spleen artery, and the infusion volume and blood loss rate were recorded; subsequently, the rats received LR resuscitation with twice the volume of blood loss. Then, blood gas indicators of the mesaconate+LR group were measured at different time points. Survival rates, indicators related to sublingual microcirculatory perfusion, liver and kidney blood flow, indicators related to the function of vital organs, and lung and brain water content were observed in all groups. RESULTS LR infusion alone could effectively maintain mean arterial pressure (MAP) within 50-60 mmHg for approximately 1 h. The administration of mesaconate combined with LR during hypotensive resuscitation could maintain MAP within 50-60 mmHg for over 3 h, with significantly reduced fluid infusion volume and blood loss rate in 50 mg/kg mesaconate+LR group, compared to the LR group ( P <0.05). In the LR group, rats maintained low pressure for up to 1 hour with a survival rate of 52.94%, and no rats survived beyond 2 h. In the 5, 20 and 50 mg/kg mesaconate+LR groups, rats maintained low pressure for up to 1 h with a survival rate exceeding 80%; in the 20 and 50 mg/kg mesaconate+LR groups, rats maintained low pressure for up to 3 h with a survival rate exceeding 70%. After complete resuscitation with mesaconate combined with LR, the 72 h survival rate of rats was 43.75%, and significant improvements in blood gas parameters were observed compared to the end of the shock phase ( P <0.05). Compared to the shock group, the mesaconate+LR group showed significant recovery in sublingual microcirculatory indicators, and liver/kidney blood flow after complete resuscitation ( P <0.05), with significant reductions in heart, liver and kidney function-related indicators and lung water content ( P <0.05). CONCLUSIONS Mesaconate combined with LR significantly extends the golden treatment time window for hemorrhagic shock in rats under high-altitude conditions, improves blood gas parameters, sublingual microcirculatory perfusion, and liver/kidney blood flow, mitigates vital organ impairment and pulmonary edema, and increases the survival rate of shocked rats.
5.Preparation of Triptolide-Chuanxiong Rhizoma Extract Ethanol Transfersomes and Analysis on Its in Vitro Anti-inflammatory Mechanism
Ling TAO ; Zhiyan WAN ; Yidan LIU ; Zhe LI ; Zhenzhong ZANG ; Weifeng ZHU ; Yongmei GUAN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):210-218
ObjectiveTo prepare triptolide-Chuanxiong Rhizoma extract ethanol transfersomes(TP-CX@TESs), conduct its quality evaluation, and investigate its in vitro anti-inflammatory efficacy and the underlying mechanisms. MethodsTP-CX@TESs was prepared via the ultrasonic injection method. With encapsulation efficiency and particle size as evaluation indicators, Box-Behnken design-response surface methodology(BBD-RSM) was employed to optimize the formulation process. The TP-CX@TESs prepared under the optimal process was characterized and evaluated for in vitro transdermal performance. A lipopolysaccharide(LPS)-induced RAW264.7 cell inflammation model was established. After 24 h of drug intervention, the levels of inflammatory factors such as nitric oxide(NO), interleukin-6(IL-6), and tumor necrosis factor-α(TNF-α) in the cell supernatant were detected. Western blot was used to determine the protein expression levels of Janus kinase 2(JAK2), signal transducer and activator of transcription 3(STAT3), and α7 nicotinic acetylcholine receptor(α7nAChR), and real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) was applied to measure the mRNA expression levels of JAK2, STAT3, the encoding gene of α7nAChR(CHRNA7), and nuclear transcription factor-κB(NF-κB). ResultsResults of BBD-RSM showed that the optimal formulation for preparing TP-CX@TESs was as follows:egg yolk lecithin content of 2.3%, ethanol volume fraction of 30%, and ratio of polysorbate-80 to egg yolk lecithin of 2∶5. Microscopic characterization revealed that TP-CX@TESs exhibited a spherical-like structure with a particle size of (105.60±3.85) nm, a polydispersity index of 0.19±0.03, and a Zeta potential of (-15.89±0.98) mV. The encapsulation efficiencies of triptolide, ferulic acid, and ligustilide were (76.88±4.40)%, (78.84±4.40)%, and (65.88±0.06)%, respectively. Both in vitro release and transdermal penetration of triptolide, ferulic acid, and ligustilide in TP-CX@TESs all followed the first-order kinetic model, showing a certain sustained-release property. Experimental results in RAW264.7 cells indicated that TP-CX@TESs significantly inhibited the release of NO, TNF-α, and IL-6(P<0.01), remarkably upregulated the protein expression levels of STAT3 and α7nAChR(P<0.01), increased the mRNA expression level of CHRNA7, and significantly downregulated the mRNA expression level of NF-κB(P<0.05, P<0.01). ConclusionThe optimized formulation process of TP-CX@TESs is simple and feasible, along with favorable in vitro release property, good transdermal permeability, and excellent in vitro anti-inflammatory activity, the mechanism is related to the inhibition of NF-κB.
6.Preparation of Triptolide-Chuanxiong Rhizoma Extract Ethanol Transfersomes and Analysis on Its in Vitro Anti-inflammatory Mechanism
Ling TAO ; Zhiyan WAN ; Yidan LIU ; Zhe LI ; Zhenzhong ZANG ; Weifeng ZHU ; Yongmei GUAN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):210-218
ObjectiveTo prepare triptolide-Chuanxiong Rhizoma extract ethanol transfersomes(TP-CX@TESs), conduct its quality evaluation, and investigate its in vitro anti-inflammatory efficacy and the underlying mechanisms. MethodsTP-CX@TESs was prepared via the ultrasonic injection method. With encapsulation efficiency and particle size as evaluation indicators, Box-Behnken design-response surface methodology(BBD-RSM) was employed to optimize the formulation process. The TP-CX@TESs prepared under the optimal process was characterized and evaluated for in vitro transdermal performance. A lipopolysaccharide(LPS)-induced RAW264.7 cell inflammation model was established. After 24 h of drug intervention, the levels of inflammatory factors such as nitric oxide(NO), interleukin-6(IL-6), and tumor necrosis factor-α(TNF-α) in the cell supernatant were detected. Western blot was used to determine the protein expression levels of Janus kinase 2(JAK2), signal transducer and activator of transcription 3(STAT3), and α7 nicotinic acetylcholine receptor(α7nAChR), and real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) was applied to measure the mRNA expression levels of JAK2, STAT3, the encoding gene of α7nAChR(CHRNA7), and nuclear transcription factor-κB(NF-κB). ResultsResults of BBD-RSM showed that the optimal formulation for preparing TP-CX@TESs was as follows:egg yolk lecithin content of 2.3%, ethanol volume fraction of 30%, and ratio of polysorbate-80 to egg yolk lecithin of 2∶5. Microscopic characterization revealed that TP-CX@TESs exhibited a spherical-like structure with a particle size of (105.60±3.85) nm, a polydispersity index of 0.19±0.03, and a Zeta potential of (-15.89±0.98) mV. The encapsulation efficiencies of triptolide, ferulic acid, and ligustilide were (76.88±4.40)%, (78.84±4.40)%, and (65.88±0.06)%, respectively. Both in vitro release and transdermal penetration of triptolide, ferulic acid, and ligustilide in TP-CX@TESs all followed the first-order kinetic model, showing a certain sustained-release property. Experimental results in RAW264.7 cells indicated that TP-CX@TESs significantly inhibited the release of NO, TNF-α, and IL-6(P<0.01), remarkably upregulated the protein expression levels of STAT3 and α7nAChR(P<0.01), increased the mRNA expression level of CHRNA7, and significantly downregulated the mRNA expression level of NF-κB(P<0.05, P<0.01). ConclusionThe optimized formulation process of TP-CX@TESs is simple and feasible, along with favorable in vitro release property, good transdermal permeability, and excellent in vitro anti-inflammatory activity, the mechanism is related to the inhibition of NF-κB.
7.Advances in molecular genetic research on Myelodysplastic syndrome.
Tao WU ; Wenhui LIU ; Yang LIU ; Qiuyue WU
Chinese Journal of Medical Genetics 2026;43(4):307-311
Myelodysplastic syndrome (MDS) is a chronic hematologic disorder characterized by ineffective hematopoiesis, dysplasia of one or more cell lines with or without definite genetic changes. Its diagnosis requires a comprehensive analysis combining morphology, immunology, cytogenetics, and molecular biology findings. In recent years, the development of second-generation sequencing (NGS) has provided great assistance in exploring the molecular pathogenesis of hematological malignancies and guidance for clinical practice. Mutations of a series of gene involved in RNA splicing, DNA methylation, transcriptional regulation, signal transduction, chromatin modification and cohesin complex have been identified as important mechanisms for the development of MDS, among which some mutations have been found to play important roles in the diagnosis, treatment, and prognosis of MDS. This article has provided a comprehensive review the the common molecular genetic abnormalities involved in MDS.
Humans
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Myelodysplastic Syndromes/diagnosis*
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Mutation
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DNA Methylation
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RNA Splicing
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High-Throughput Nucleotide Sequencing
8.Surveillance of schistosomiasis in Jiangsu Province from 2012 to 2024
Wei LI ; Jianfeng ZHANG ; Liang SHI ; Tao WANG ; Yun FENG ; Lu LIU ; Kun YANG
Chinese Journal of Schistosomiasis Control 2026;38(1):8-13
Objective To evaluate the effectiveness of schistosomiasis surveillance in Jiangsu Province during the stage moving from transmission control to transmission interruption, and to analyze the current risk and challenges, so as to provide the evidence for achieving the target of schistosomiasis elimination. Methods Schistosomiasis surveillance data were collected from Jiangsu Province from 2012 to 2024, and the endemic areas, Schistosoma japonicum infections in humans and livestock, Oncomelania hupensis snail distribution and implementation of integrated interventions were descriptively analyzed. In addition, the trends in areas with snails, seroprevalence of human S. japonicum infections and numbers of advanced schistosomiasis cases were assessed using a Joinpoint regression model. Results The endemic areas of schistosomiasis continued to shrink in Jiangsu Province from 2012 to 2024, with the number of schistosomiasis-eliminated counties (cities, districts) increasing from 53 (75.71%) to 63 (96.92%), and interruption of schistosomiasis transmission was achieved across the province. A total of 4 600 300 person-times were tested for serum antibodies against S. japonicum, with 28 719 person-times positive detected; and 616 500 person-times were tested S. japonicum infections among local residents in Jiangsu Province from 2012 to 2024, with only 3 egg-positives detected, and no egg-positives found since 2017. A total of 187 600 herd-times were tested for schistosomiasis in livestock, and no S. japonicum infections were found. O. hupensis snail survey was performed covering 1 018 408.97 hm2, and a total of 35 556.35 hm2 was found with snail-infested habitats, including 174.40 hm2 of emerging snail-infested habitats. A total of 1 102 800 O. hupensis snails were identified for S. japonicum infections, and no infections were found. The areas of snail-infested habitats appeared a tendency towards a rise in Jiangsu Province from 2019 to 2023 (APC = 23.67%, P < 0.05), and the actual areas of snail-infested habitats appeared a tendency towards a decline from 2012 to 2015 (APC = −22.77%, P < 0.05), and towards a rise from 2015 to 2023 (APC = 9.76%, P < 0.01). The seroprevalence of anti-S. japonicum antibodies appeared a tendency towards a decline among residents in Jiangsu Province from 2017 to 2023 (APC = −14.92%, P < 0.01). In addition, the number of newly diagnosed advanced schistosomiasis cases appeared a tendency towards a decline from 2012 to 2024 (APC = −12.02%, P < 0.01), and the numbers of advanced schistosomiasis patients requiring treatment showed a tendency towards a decline from 2012 to 2021 (APC = −10.56%, P < 0.01) and from 2021 to 2023 (APC = −20.06%, P < 0.01). Conclusions Great progresses had been achieved in schistosomiasis control in Jiangsu Province following transmission control, and transmission interruption had been achieved; however, there are still snail-infested habitats. High-intensity surveillance and integrated control are required to be maintained to advance the achievement of the target of schistosomiasis elimination in Jiangsu Province.
9.Polypeptide-based Nanocarriers for Oral Targeted Delivery of CAR Genes to Pancreatic Cancer
Feng XIN ; Jian REN ; Zhao-Zhen LI ; Quan FANG ; Rui-Jing LIANG ; Lan-Lan LIU ; Lin-Tao CAI
Progress in Biochemistry and Biophysics 2026;53(2):431-441
ObjectivePancreatic ductal adenocarcinoma (PDAC) exhibits a limited response to current treatments due to its dense fibrotic stroma and highly immunosuppressive tumor microenvironment. In recent years, advancements in cellular immunotherapy, particularly chimeric antigen receptor macrophage (CAR-M) therapy, have offered new hope for pancreatic cancer treatment. Although CAR-M therapy demonstrates dual potential in directly killing tumor cells and remodeling the immune microenvironment, it still faces challenges such as complex in vitro preparation processes and low in vivo targeting and delivery efficiency. Therefore, developing strategies for efficient and targeted in vivo delivery of CAR genes has become crucial for overcoming current therapeutic limitations. This study aims to develop an orally administrable nano-gene delivery system for the targeted delivery of CAR genes to pancreatic tumor sites. MethodsCore nano-gene particles (PNP/pCAR) were constructed by loading plasmid DNA encoding CAR (pCAR) with cationic polypeptides (PNP). Subsequently, PNP/pCAR was surface-modified with β-glucan to prepare the targeted nanoparticles (βGlus-PNP/pCAR). The loading efficiency of PNP for pCAR was quantitatively assessed by gel retardation assay. The particle size, Zeta potential, morphology, and storage stability of PNP/pCAR were characterized using a Malvern particle size analyzer and transmission electron microscopy. At the cellular level, RAW 264.7 macrophages were selected. The cytotoxicity of PNP/pCAR was evaluated using the CCK-8 assay. The cellular uptake efficiency and lysosomal escape ability of the nanoparticles were assessed via flow cytometry and confocal microscopy. Transfection efficiency was quantitatively evaluated by detecting the expression of the reporter gene GFP using flow cytometry. At the in vivo level, an orthotopic pancreatic cancer mouse model was established. Cy7-labeled βGlus-PNP/pCAR nanoparticles were administered orally, and the fluorescence distribution in mice was dynamically monitored at 1, 2, 4, 8, and 16 h post-administration using a small animal in vivo imaging system. Forty-eight hours after oral gavage, the mice were euthanized, and pancreatic tumor tissues were collected for further analysis of intratumoral fluorescence signals using the imaging system. Additionally, βGlus-PNP/pCAR-GFP nanoparticles loaded with the reporter gene (GFP) were administered orally. Forty-eight hours post-administration, pancreatic tumor tissues were harvested to prepare frozen sections, and GFP expression was observed and analyzed under a fluorescence microscope. ResultsThe PNP carrier exhibited a high loading capacity for pCAR. The successfully prepared PNP/pCAR nanoparticles were regular spheres with a hydrodynamic diameter of approximately (120±10) nm and a Zeta potential of about +(6±1) mV. They maintained good structural stability after incubation in PBS buffer for 7 d. Cell experiments demonstrated that PNP/pCAR exhibited no significant cytotoxicity in RAW 264.7 cells while being efficiently internalized and effectively escaping lysosomal degradation. The transfection positive rate of PNP/pCAR-GFP in RAW 264.7 cells reached (25±3)%, surpassing that of Lipofectamine 2000-loaded pCAR-GFP (Lipo/pCAR-GFP), which was (20±1)%.In vivo experiments revealed that, compared to unmodified PNP/pCAR, βGlus-PNP/pCAR exhibited strongerin situ pancreatic tumor targeting ability after oral administration. Furthermore, oral administration of βGlus-PNP/pCAR-GFP resulted in significant GFP protein expression detectable within pancreatic tumor tissues. ConclusionThis study successfully constructed and validated an orally administrable, pancreatic cancer-targeting polypeptide-based nano-gene delivery system. It provides an important technological foundation in delivery systems and experimental basis for the subsequent development of in situ CAR-M-based therapeutic strategies for pancreatic cancer.
10.Polypeptide-based Nanocarriers for Oral Targeted Delivery of CAR Genes to Pancreatic Cancer
Feng XIN ; Jian REN ; Zhao-Zhen LI ; Quan FANG ; Rui-Jing LIANG ; Lan-Lan LIU ; Lin-Tao CAI
Progress in Biochemistry and Biophysics 2026;53(2):431-441
ObjectivePancreatic ductal adenocarcinoma (PDAC) exhibits a limited response to current treatments due to its dense fibrotic stroma and highly immunosuppressive tumor microenvironment. In recent years, advancements in cellular immunotherapy, particularly chimeric antigen receptor macrophage (CAR-M) therapy, have offered new hope for pancreatic cancer treatment. Although CAR-M therapy demonstrates dual potential in directly killing tumor cells and remodeling the immune microenvironment, it still faces challenges such as complex in vitro preparation processes and low in vivo targeting and delivery efficiency. Therefore, developing strategies for efficient and targeted in vivo delivery of CAR genes has become crucial for overcoming current therapeutic limitations. This study aims to develop an orally administrable nano-gene delivery system for the targeted delivery of CAR genes to pancreatic tumor sites. MethodsCore nano-gene particles (PNP/pCAR) were constructed by loading plasmid DNA encoding CAR (pCAR) with cationic polypeptides (PNP). Subsequently, PNP/pCAR was surface-modified with β-glucan to prepare the targeted nanoparticles (βGlus-PNP/pCAR). The loading efficiency of PNP for pCAR was quantitatively assessed by gel retardation assay. The particle size, Zeta potential, morphology, and storage stability of PNP/pCAR were characterized using a Malvern particle size analyzer and transmission electron microscopy. At the cellular level, RAW 264.7 macrophages were selected. The cytotoxicity of PNP/pCAR was evaluated using the CCK-8 assay. The cellular uptake efficiency and lysosomal escape ability of the nanoparticles were assessed via flow cytometry and confocal microscopy. Transfection efficiency was quantitatively evaluated by detecting the expression of the reporter gene GFP using flow cytometry. At the in vivo level, an orthotopic pancreatic cancer mouse model was established. Cy7-labeled βGlus-PNP/pCAR nanoparticles were administered orally, and the fluorescence distribution in mice was dynamically monitored at 1, 2, 4, 8, and 16 h post-administration using a small animal in vivo imaging system. Forty-eight hours after oral gavage, the mice were euthanized, and pancreatic tumor tissues were collected for further analysis of intratumoral fluorescence signals using the imaging system. Additionally, βGlus-PNP/pCAR-GFP nanoparticles loaded with the reporter gene (GFP) were administered orally. Forty-eight hours post-administration, pancreatic tumor tissues were harvested to prepare frozen sections, and GFP expression was observed and analyzed under a fluorescence microscope. ResultsThe PNP carrier exhibited a high loading capacity for pCAR. The successfully prepared PNP/pCAR nanoparticles were regular spheres with a hydrodynamic diameter of approximately (120±10) nm and a Zeta potential of about +(6±1) mV. They maintained good structural stability after incubation in PBS buffer for 7 d. Cell experiments demonstrated that PNP/pCAR exhibited no significant cytotoxicity in RAW 264.7 cells while being efficiently internalized and effectively escaping lysosomal degradation. The transfection positive rate of PNP/pCAR-GFP in RAW 264.7 cells reached (25±3)%, surpassing that of Lipofectamine 2000-loaded pCAR-GFP (Lipo/pCAR-GFP), which was (20±1)%.In vivo experiments revealed that, compared to unmodified PNP/pCAR, βGlus-PNP/pCAR exhibited strongerin situ pancreatic tumor targeting ability after oral administration. Furthermore, oral administration of βGlus-PNP/pCAR-GFP resulted in significant GFP protein expression detectable within pancreatic tumor tissues. ConclusionThis study successfully constructed and validated an orally administrable, pancreatic cancer-targeting polypeptide-based nano-gene delivery system. It provides an important technological foundation in delivery systems and experimental basis for the subsequent development of in situ CAR-M-based therapeutic strategies for pancreatic cancer.


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