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.

Objective: Scutellarin in Shuganning injection has potential allergenicity and is prone to causing anaphylactoid reactions. There is an urgent need to establish a simple and rapid chromatographic quantitative method to achieve the limit control of scutellarin in Shuganning injection.
Methods: For scutellarin, HPLC method was used in the quantitative analysis using a Shim-pack GIS C₁₈ (2.1 mm×150 mm, 5 μm) column. The mobile phase was a blend of 0.1% trifluoroacetic acid in water and acetonitrile, and gradient elution progress was applied for the whole process. The flow rate was 0.3 mL·min^-1 and the column temperature was maintained at 37 ℃. The detecting wavelength was 320 nm.
Results: The linear regression equation was y=51 456x-27 352, the correlation coefficient r=0.999 8, namely the calibration curve was linear in the range of 1.02-102.00 μg·mL^-1; the average recovery was 103.76%; the detection and quantification limit for determining scutellarin was 0.16 μg·mL^-1 and 0.57 μg·mL^-1, respectively; the injection sample was stable at room temperature for 48 h.
Conclusion: This method is easy to operate, reproducible and has high accuracy, thus can be used for quality control of Shuganning injection.

Objective:To explore the influence of value orientation brief therapy (VBT) on anxiety and depression symptoms, social function, coping style and self-acceptance level of major depressive disorder adolescents with anxious distress.Methods:From June 2021 to June 2022, seventy adolescent major depressive disorder patients with anxious distress were included in the study, who were randomly divided into study group(35 people, 31 people completed)and control group (35 people, 30 people completed). The study group was given routine treatment combined with VBT, while the control group was given routine treatment only. Before and after treatment, Hamilton anxiety scale(HAMA), Hamilton depression scale (HAMD), social disability screening schedule (SDSS), coping style questionnaire(CSQ) and self-acceptance questionnaire (SAQ) were used to evaluate the two groups, and SPSS 26.0 software was used to statistically analyze the data of the two groups. Paired sample t-test was used for intra-group comparison, and independent sample t-test was used for inter-group comparison. Results:After 6 weeks of treatment, the scores of HAMA((6.03±3.58) vs (14.03±7.06), t=5.55, P<0.01), HAMD((8.77±5.52 ) vs (16.50±7.59), t=4.56, P<0.01)and SDSS((4.23±1.50) vs (6.63±0.96), t=7.43, P<0.01)in the study group were significantly lower than those in the control group, the differences were statistically significant. The scores of self-acceptance((19.23±1.33) vs (13.47±1.46), t=-16.12, P<0.01)and self-evaluation ((19.87±2.87) vs (12.77±1.68), t=-11.75, P<0.01) in the SAQ scale and the scores of problem-solving((8.71±2.30) vs (6.23±3.45), t=3.31, P<0.05) and rationalization ((6.20±3.11) vs (4.67±2.43), t=2.13, P<0.05) in the CSQ questionnaire were significantly higher than those in the control group, the differences were statistically significant. The total effective rate of the study group(90.3%(28/31) vs 66.7%(20/30), χ2=5.09, P<0.05) was significantly higher than that of the control group, the difference was statistically significant. Conclusion:The effect of routine treatment combined with VBT is better, which can effectively improve anxiety and depression symptoms, social function and coping style, and enhance self-acceptance and self-evaluation in adolescent major depressive disorder patients, which is worthy of clinical application.

In recent years,organs-on-chips(OOCs)technology has gradually matured and been recognized by regulatory agencies,playing a key role in the construction of pathophysiological models,pharmacodynamic screening,toxicological evaluation and other fields,providing guidance for the research and development of innovative drugs of Chinese materia medica and the clinical application of classic prescriptions.This article reviewed the application progress of OOCs technology in the research of Chinese materia medica from four dimensions:pharmacodynamic and toxicological evaluation,quality control,active component analysis and innovative drug research and development,in order to provide a reference for the precise application of this technology in the field of Chinese materia medica.

The purpose of this study is to develop a shark single domain antibody(SdAb)targeting a unique B cell epitope in the SARS-CoV-2 spike protein,and explore its role in the immunological detection targeting SARS-CoV-2 spike protein S.A u-nique peptide S9 was artificially synthesized based on the sequence of a unique B cell epitope of SARS-CoV-2 spike protein,then it was conjugated to the carrier protein KLH.It was used as an immunogen for subcutaneous injection into shark back and boosted according to the standard immunization protocol.Blood collected from shark tail vein and peripheral blood lymphocytes(PBL)were isolated.Total RNA was purified from PBL and transcribed to cDNA by reverse transcription.Shark vNAR frag-ments were amplified from cDNA templates and cloned into pComb3XSS vector to obtain phage library.A positive clone named T01 was obtained through screening the phage library by indirect ELISA.Then its gene was cloned into the expression vector pET-28a.The SdAb T01 was then prokaryotically expressed and purified,and its specific recognition of the SARS-CoV-2 spike protein S was indentified by Western-blot(WB),indirect ELISA and IF A.T01 binds well with peptide S9 at EC50 value of 2.050±0.064 nmol/L.The purified SdAb T01 was proven by WB to be able to selectively detect recombinant spike protein sub-unit 1(S1)of SARS-CoV-2,with no cross-reactive to recombinant spike protein subunit 1 of other six human coronavirus.It was showed by ELISA that SdAb T01 can sensitively detect the recombinant N terminal domain(NTD)of SARS-CoV-2 pro-tein.Moreover,it also specifically recognizes the spike protein of SARS-CoV-2 that was transiently expressed in transfected HEK293 cells by IFA.Therefore,a shark single domain antibody targeting a unique B cell epitope in the spike protein of SARS-CoV-2 was successfully developed,and has shown potential immunodiagnostic value by WB,ELISA and IFA.Thus,it provides an effective tool for unique antigen detection of SARS-CoV-2.

Endometriosis(EMs)is a common estrogen-depend-ent clinical disease with the pathological characteristics of malig-nant tumors,which has great impact on women's physical and mental health.In recent years,experimental exploration has re-vealed that ectopic foci are in a hypoxic environment outside the uterine cavity,and mitochondria,as the"functional factories"of the cells,play an important role in the process of planting and in-vasion,and the mitochondrial quality control system,which in-cludes mitochondrial oxidative stress,kinetics,autophagy,bio-genesis and calcium homeostasis,is a key mechanism for the e-quilibrium of the mitochondrial function.The mitochondrial quality control system,including mitochondrial oxidative stress kinetics,autophagy,biogenesis and calcium homeostasis,is a key mechanism for mitochondrial functional balance.Therefore,to clarify the role of the mitochondrial quality control system in the development of EMs with the help of rational and rigorous experi-mental and clinical studies can not only help to clarify the patho-genesis of the disease,but also explore the key targets in the prevention and treatment of the disease.Therefore,this article summarizes the research progress of mitochondrial quality control system in endometriosis,with a view to providing reference and theoretical basis for the etiology,pathogenesis and prevention strategies of EMs.

Three common faults of Siemens SOMATOM Force Dual Source CT were investigated in terms of phenomenon,cause,troubleshooting measure and maintenance experience.References were provided for treating similar faults.[Chinese Medical Equipment Journal,2025,46(9):114-117]

The composition and working principle of MAQUET ROTAFLOW ECMO system were introduced.Four cases of common failures of ROTAFLOW ECMO systems were analyzed in terms of the cause and elimination method.The methods for the maintenance and quality control of ECMO system were put forward.References were provided for engineering technicians to treat similar faults.[Chinese Medical Equipment Journal,2025,46(5):116-120]

Objective:To construct an evaluation index system for the operational management capability in tertiary public hospitals and validate its feasibility,providing a reference for accurately assessing the operational management capability of public hospitals.Methods:A literature review and Delphi method were employed to construct the index system.The Analytic Hierarchy Process was used to calculate weights,and a weighted comprehensive index method was applied to evaluate the operational management capability of sample hospitals.Results:A hierarchical indicator system was established,comprising 6 first-level indicators,16 second-level indicators,and 40 third-level indicators,covering the dimensions of resource allocation,service efficiency,economic operation,risk prevention and control,development capacity,and social benefits.Empirical results indicated that the composite score of the sample hospital decreased from 71.5 in 2019 to 53.8 in 2020 and rebounded to 57.9 by 2023.Conclusions:The empirical outcomes align well with the operational capacity as understood by the operational management personnel of the sample hospital,demonstrating that the indicator system is scientifically sound and rational,possessing a high evaluative capability.