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.

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.

ObjectiveTo establish a castrated male mouse model and to preliminarily investigate the effects of testosterone replacement therapy （TRT） on behavior， serum indices， and histopathological changes in castrated mice， as well as to explore the role of androgens in cognitive function. MethodsForty 6-month-old male C57/BL6J mice were randomly divided into sham operation group， castration group， testosterone propionate （0.5，1.0 mg/kg） treated group， with 10 mice in each group. Following castration and subcutaneous administration of testosterone propionate at different doses （0.5 and 1.0 mg/kg） for TRT， learning and memory abilities were assessed using the Morris water maze （MWM） test and the passive avoidance test. Serum testosterone and serum brain-derived neurotrophic factor （BDNF） levels were measured by ELISA， and histopathological changes in the hippocampus were examined using hematoxylin-eosin （HE） staining. ResultsRoutine observations： there were no statistically significant differences in body weight among groups at any time point. MWM test： compared with castration group， sham operation group and testosterone propionate-treated groups （0.5， 1.0 mg/kg） showed significantly reduced escape latency on days 4 and 5 （P0.05）， while the number of platform crossings and the time spent in the target quadrant significantly increased （P0.05）. Passive avoidance test： the number of passive avoidance errors significantly decreased in sham operation group and testosterone propionate （1.0 mg/kg）-treated group （P0.05）， and the passive avoidance latency was significantly prolonged in sham-operated group and testosterone propionate-treated groups （0.5， 1.0 mg/kg） （P0.05）. Serum testosterone and serum BDNF assays： serum testosterone levels and serum BDNF concentrations significantly increased in sham operation group and testosterone propionate-treated groups （0.5， 1.0 mg/kg） （P0.01）. HE staining： compared with sham operation group， neuronal density in all hippocampal subregions was slightly reduced in castration group； in the testosterone propionate （0.5 mg/kg）-treated group， neuronal arrangement in the CA1 and CA3 regions was improved and apoptotic cells were reduced compared with castration group； in testosterone propionate （1.0 mg/kg）-treated group， the pyramidal cell layer in the CA3 region was more compactly arranged， with fewer apoptotic cells than in castration group. ConclusionTRT improves learning and memory performance in castration male mice， potentially through modulation of hippocampal BDNF signaling pathways.

ObjectiveTo construct a nomogram prediction model for non-valvular persistent atrial fibrillation （PeAF） patients with left ventricular hypertrophy （LVH） ， followed by prognostic analysis through follow-up. MethodsThis study retrospectively enrolled 949 patients with newly diagnosed and hospitalized non-valvular PeAF. Among them， 403 patients presented with LVH. The cohort was randomly stratified into a training set （n=665） and a validation set （n=284）. Univariate and multivariate Logistic regression analyses were employed to identify independent risk factors for PeAF complicated by LVH. A nomogram prediction model was subsequently constructed and evaluated for discriminative ability， calibration， and clinical utility using receiver operating characteristic （ROC） curve analysis， calibration plots， and decision curve analysis （DCA）. ResultsSeven independent risk factors were ultimately identified and included in the prediction model： female sex， hypertension， diabetes， red blood cell distribution width-SD （RDW-SD）， body mass index （BMI）， left atrial diameter （LAD）， and left ventricular ejection fraction （LVEF）. The area under the ROC curve （AUC） in the training set was 0.862 （95% CI： 0.834-0.890）， and in the validation set， it was 0.870 （95% CI： 0.829-0.911）， demonstrating excellent predictive performance. ConclusionIndependent risk factors for LVH in PeAF patients include female， hypertension， diabetes， RDW-SD， BMI， LAD， and LVEF. The prediction model built based on this can help early identification of PeAF patients with high risk of LVH. At the same time， the incidence of major adverse cardiovascular events （MACE） is higher in PeAF patients with LVH. Patients with atrial fibrillation combined with LVH may benefit from catheter ablation.

ObjectiveTo investigate the possible mechanism of Pibai Yucuo Formula （枇柏愈痤方， PYF） in treating acne from the perspective of skin barrier damage. MethodsThirty-two mice were randomly divided into blank group， model group， minocycline group， and PYF group， with 8 mice in each group. Except for the blank group， mice were induced by intradermal injection of Cutibacterium acnes （C.acnes） combined with topical application of artificial sebum to establish acne model. The blank group and model group received intragastric administration of 0.2 ml of distilled water， while the PYF group received intragastric administration of 22.75 g/（kg·d）of PYF， and the minocycline group received 0.013 g/（kg·d）of minocycline suspension， all once daily for 5 consecutive days. On day 0 and day 6 of the experiment， the body weight of mice in each group was recorded， and the absolute value of the body weight difference during the experiment was calculated. Skin conditions were assessed with multifunctional skin imaging system on the 2nd， 4th and 6th day of the experiment. Skin barrier function indicators including transepidermal water loss （TEWL）， and the water content of the stratum corneum and epidermis on day 0， 2， 4 and 6 of the experiment. Optical coherence tomography （OCT） was used to observe stratum corneum and skin thickness on the 1st， 3rd and 5th day of the experiment. Hematoxylin-eosin （HE） staining was performed to observe histopathological changes， while ELISA was used to detect interleukin-17A （IL-17A） levels， and immunofluorescence staining was used to assess skin barrier-related proteins filaggrin （FLG） and loricrin （LOR） levels of skin lesions on day 6 of the experiment. ResultsCompared to the blank group， the model group showed a decrease in body weight on day 6， and an increase in the absolute value of the difference in body weight before and after the experiment （P＜0.05）. On day 4 and 6， TEWL values increased， while water content in the skin stratum corneum and epidermis decreased （P＜0.05）， accompanied by elevated IL-17A level and reduced immunofluorescence intensity of FLG and LOR proteins （P＜0.05）. The model group mice showed papules or pustules at the skin modeling site with progressively worsening desquamation under multifunctional skin imaging system. OCT revealed focal epidermal protrusions， blurred epidermal-dermal boundaries， and disorganized structural layers. HE staining showed significant epidermal hyperkeratosis and incomplete keratinization in the skin， with keratin plug formation in hair follicles and glandular lumens， thickened stratum corneum， hyperplasia of the stratum spinosum， as well as dense dermal inflammatory cell infiltration， and capillary dilation. Compared to the model group， both the minocycline group and the PYF group showed a reduced difference in body weight before and after experiment （P＜0.05）. On day 4 and 6， the TEWL value decreased， and water content of the skin stratum corneum increased （P＜0.05）； on day 6， the IL-17A level in the skin lesions decreased and immunofluorescence intensity of FLG and LOR proteins increased （P＜0.05）. On day 4 and 6， the severity of the skin lesions and range of redness and swelling were lighter than those in the model group， with reverted epidermal thickness， smoother surface and clearer epidermis-dermis boundary. HE staining showed that the degree of skin keratinization was reduced， and the inflammatory infiltration and vascular dilation in the dermis were improved compared to the model group. The PYF group showed better results than the minocycline group in reducing TEWL value on day 4 （P＜0.05）. ConclusionPYF may improve inflammation and skin barrier damage by downregulating IL-17A levels in lesion tissue and increasing skin barrier-related proteins， which could be one of the potential mechanism of action on acne.

Objective:To analyze and summarize the clinical features and short-medium term follow-up results of children with multisystem inflammatory syndrome（MIS-C）following coronavirus infection.Methods:The data of six children with MIS-C admitted to the Intensive Care Unit of Shanghai Children's Medical Center from January to March 2023 were retrospectively analyzed.Results:All six cases were in shock，requiring vasoactive drugs，and one case required invasive mechanical ventilation.All the six patients had multiple organ function injury and increased inflammation indicators.After admission，they received organ support，glucocorticoids and gamma globulin treatment.Two patients were treated with biological agents.Both organ function and inflammation indicators showed significantly improvement after therapy.Six patients had mild coronary artery widening.All patients had good prognosis following short-medium term follow-up.Conclusion:Children with severe MIS-C may suffer life-threatening hemodynamic instability.Timely assessment，active anti-inflammatory and organ support therapy can obtain favorable prognosis.

Objective To explore the method for grading the degree of uterine adhesion in a rat model.Methods A rat model of uterine adhesion was established using the double-injury method.Paraffin sections were observed using HE staining and Masson staining to compare morphological changes in the uterus,endometrial thickness,gland and vessel counts,uterine cavity area,and adhesion severity.Rat sections were classified into three grades based on uterine cavity area for comparative analysis.Results The average uterine cavity area and uterine cavity area/endometrial layer area were smaller in rats in the model group compared with the blank group(P＜0.01).The uterine cavity area/endometrial layer area ratio was categorized into grades Ⅰ,Ⅱ,and Ⅲ,with a significant difference among the grades(P＜0.05,P＜0.01).Conclusions The uterine cavity area/endometrial layer area ratio may reflect the grading difference in the degree of uterine adhesion in rats with uterine adhesions.This ratio may thus be used as a grading-evaluation criterion in the rat model of uterine adhesion,with implications for diagnostic grading in this model.

Aim To clarify the mechanism by which Qishen Yixin Granules improved microcirculation vas-cular endothelial dysfunction(VED)in mice,through activating the Nrf2/HO-1 signaling pathway to regulate oxidative stress.Methods C57 mice were randomly divided into six groups:blank group,model group,pos-itive drug group,and low-,medium-,and high-dose groups of Qishen Yixin Granules.The VED model was established by long-term infusion of Ang Ⅱ combined with a high-fat diet.Each treatment group received the corresponding drug intervention.After four weeks of drug intervention,cardiac function was assessed by echocardiography.Carstairs staining was used to ob-serve the formation of microthrombi in myocardial tis-sue.The micro vascular ischemia was evaluated by Hei-denhain staining.The ultrastructure of endothelial cells was observed by electron microscopy.The levels of EMPs,ROS,NO,ET-1,TF,TM,VWF,and TXA2 in serum were measured by ELISA.The expression levels of MDA,SOD,and GSH-Px in mouse heart tissue were determined by chemical methods.Cardiac microvascu-lar density and the expression of Nrf2,Keap1,and HO-1 proteins were detected by Immunohistochemical stai-ning.The protein expressions of Keap1,cytoplasmic Nrf2,nuclear Nrf2,and HO-1 in myocardial tissue were detected by Western blot.Results Qishen Yixin Granules could effectively improve the cardiac function of mice,alleviate the damage of endothelial cells and endothelial function.They could up-regulate serum NO levels and the activities of antioxidant enzymes SOD and GSH-Px,while down-regulating the expression of ROS and vascular inflammatory injury factors such as ET-1,VWF,TXA2,TF,TM,and EMPs.Qishen Yixin Granules also increased the positive counts of CD34,Nrf2,and HO-1,as well as microvessel density.Fur-thermore,they inhibited the expression of MDA,Keap1,and cytoplasmic Nrf2 protein in myocardial tis-sue,while increasing the expression of nuclear proteins HO-1 and Nrf2.Conclusions Qishen Yixin Granules may inhibit oxidative stress and inflammatory response by regulating the Nrf2/HO-1 signaling pathway,thereby improving vascular endothelial damage and cardiac function in VED mice.

Objective:To construct an esophageal language rehabilitation program for patients after total laryngectomy (TL) based on the behavioral change wheel theory and exploring its application effects.Methods:The multidisciplinary team constructed the first draft of the esophageal language rehabilitation program for patients after TL based on the behavioral change wheel theory, combined with a literature review, and used an expert meeting to revise the rehabilitation program to form the final draft of the program. Adopting experimental research, 35 patients after TL who attended the nursing outpatient clinic of Beijing Tongren Hospital, Capital Medical University from February 2023 to May 2023 were selected as the control group by convenience sampling method, and 35 patients after TL who attended the nursing outpatient clinic from June 2023 to September 2023 were selected as the experimental group. The experimental group applied the TL postoperative patients′ esophageal language rehabilitation program based on the behavioral change wheel theory on the basis of conventional nursing measures, and the control group received conventional nursing care. The status of language rehabilitation training, quality of life, social behavioral status, and anxiety and depression status before intervention, 1, 3, 6 months after intervention were compared between the two groups.Results:Both groups of patients completed the study. There were 29 males and 6 females in the control group, with an age of (54.63 ± 10.44) years old. There were 34 males and 1 female in the experimental group, with an age of (55.17 ± 10.67) years old. There was no statistically significant difference in the language rehabilitation training status, quality of life, social behavior status, and anxiety and depression status between the two groups before intervention (all P>0.05). The total scores of speech rehabilitation training in the experimental group at 1, 3, and 6 months after intervention were (32.80 ± 2.49), (39.80 ± 2.75), (51.91 ± 4.20) points, respectively, which were higher than those in the control group (23.40 ± 3.42), (24.40 ± 3.42), (25.80 ± 3.42) points, and the differences were statistically significant ( t=14.53, 23.44, 32.70, all P<0.05). The total scores of quality of life in the experimental group at 1, 3, and 6 months after intervention were (98.91 ± 8.49), (134.66 ± 11.31), (157.97 ± 13.97) points, respectively, which were higher than those in the control group (67.06 ± 7.64), (72.16 ± 7.64), (99.46 ± 8.09) points, and the differences were statistically significant ( t=17.53, 30.16, 21.45, all P<0.05). The scores for social occasion diet and language comprehension in the experimental group at 1, 3, and 6 months after intervention were (40.41 ± 13.94), (40.43 ± 24.08), (40.60 ± 18.56), and (43.71 ± 12.26), (47.40 ± 17.09), (52.50 ± 13.82), respectively, which were higher than those in the control group (30.59 ± 15.98), (30.57 ± 18.28), (27.21 ± 15.27), and (27.29 ± 15.13), (23.60 ± 14.78), (19.50 ± 12.78), and the differences were statistically significant ( t values were -6.88-2.16, all P<0.05). The total scores of anxiety and depression in the experimental group at 1, 3, and 6 months after intervention were (23.74 ± 2.73), (14.89 ± 3.89), (12.11 ± 3.14) points, respectively, which were lower than those in the control group (32.63 ± 1.85), (30.63 ± 1.85), (24.80 ± 2.75) points, and the differences were statistically significant (t=-19.55, -27.10, -17.97, all P<0.05). Conclusions:The esophageal language rehabilitation program for patients after TL based on the behavioral change wheel theory in this study is scientific, feasible, and can improve the patients′ esophageal language expression, quality of life, anxiety and depression, and social behavioral status, which can provide a reference for clinical care.

Objective To examine and compare the effect of personalized progressive aerobic-exercise and resistance-training prescriptions on anxiety of undergraduates.Methods This was a randomized controlled trial.Sixty-six undergraduates with anxiety were recruited and randomized into an aerobic ex-ercise(AE)group,a resistance exercise(RE)group and a control group,each of 22.The aerobic and resistance exercise groups underwent 12-week aerobic and resistance exercise respectively,while the control group only received health education.Before as well as after 4-,8-and 12-week interven-tion,and 4 weeks after the intervention,all groups were evaluated using Self-Rating Anxiety Scale(SAS)and Chinese College Students Mental Health Scale--Anxiety Subscale(CCSMHS-AS).More-over,before and 16 weeks after the intervention,all groups were assessed physical activity(PA)us-ing the International Physical Activity Questionnaire-Short Form(IPAQ-SF).Results The average SAS scores of the AE and RE groups decreased significantly from 6.95±6.32 and 56.41±5.45 before the intervention to 38.29±5.82 and 41.18±7.51 after 12-week exercise,while the average CC-SMHS-AS score decreased significantly from 20.00±5.66 and 19.41±3.70,to 13.18±4.81 and 14.32±4.16 during the same period of time(P<0.01 for all).Four weeks after the intervention,the SAS score of the AE group was significantly higher than 4 weeks earlier(49.18±11.84 vs.38.29±5.82,P<0.01),while that of the RE group increased without significant differences compared with 4 weeks earlier(42.50±9.57 vs.41.18±7.51,P>0.05),with the value of both groups significantly lower than right after the intervention(P<0.01,P<0.05).In the control group,the SAS score de-creased significantly from 55.73±5.27 before the intervention to 47.09±5.55 right after the interven-tion,and further to 46.95±9.70 4 weeks later(P<0.05),but no significant differences were ob-served in the CCSMHS-AS score(P>0.05).Meanwhile,right after the intervention,the average SAS scores of the AE and RE groups were significantly lower than the control group(P<0.01,P<0.05),without significant differences among the three groups 4 weeks after the intervention(P>0.05).The CC-SMHS-AS scores of AE group right after and 4 weeks after the intervention were significantly higher than the control group(P<0.01),but no significant differences were found in it between either the AE and RE group,or the RE and control group(P>0.05).Besides,the PA levels of the AE and RE groups 4 weeks after the intervention were significantly higher than before the intervention,while no significant changes were observed in the PA level of the control group(P>0.05).Conclusion Twelve-week personalized progressive aerobic-exercise and resistance-training prescriptions both result in a similar effect on relieving anxiety and improving spontaneous PA of college students.However,the prognosis of aerobic exercise is poorer than the other.