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.

Collagen is a major structural protein in the matrix of animal cells and the most widely distributed and abundant functional protein in mammals. Collagen’s good biocompatibility, biodegradability and biological activity make it a very valuable biomaterial. According to the source of collagen, it can be broadly categorized into two types: one is animal collagen; the other is recombinant collagen. Animal collagen is mainly extracted and purified from animal connective tissues by chemical methods, such as acid, alkali and enzyme methods, etc. Recombinant collagen refers to collagen produced by gene splicing technology, where the amino acid sequence is first designed and improved according to one’s own needs, and the gene sequence of improved recombinant collagen is highly consistent with that of human beings, and then the designed gene sequence is cloned into the appropriate vector, and then transferred to the appropriate expression vector. The designed gene sequence is cloned into a suitable vector, and then transferred to a suitable expression system for full expression, and finally the target protein is obtained by extraction and purification technology. Recombinant collagen has excellent histocompatibility and water solubility, can be directly absorbed by the human body and participate in the construction of collagen, remodeling of the extracellular matrix, cell growth, wound healing and site filling, etc., which has demonstrated significant effects, and has become the focus of the development of modern biomedical materials. This paper firstly elaborates the structure, type, and tissue distribution of human collagen, as well as the associated genetic diseases of different types of collagen, then introduces the specific process of producing animal source collagen and recombinant collagen, explains the advantages of recombinant collagen production method, and then introduces the various systems of expressing recombinant collagen, as well as their advantages and disadvantages, and finally briefly introduces the application of animal collagen, focusing on the use of animal collagen in the development of biopharmaceutical materials. In terms of application, it focuses on the use of animal disease models exploring the application effects of recombinant collagen in wound hemostasis, wound repair, corneal therapy, female pelvic floor dysfunction (FPFD), vaginal atrophy (VA) and vaginal dryness, thin endometritis (TE), chronic endometritis (CE), bone tissue regeneration in vivo, cardiovascular diseases, breast cancer (BC) and anti-aging. The mechanism of action of recombinant collagen in the treatment of FPFD and CE was introduced, and the clinical application and curative effect of recombinant collagen in skin burn, skin wound, dermatitis, acne and menopausal urogenital syndrome (GSM) were summarized. From the exploratory studies and clinical applications, it is evident that recombinant collagen has demonstrated surprising effects in the treatment of all types of diseases, such as reducing inflammation, promoting cell proliferation, migration and adhesion, increasing collagen deposition, and remodeling the extracellular matrix. At the end of the review, the challenges faced by recombinant collagen are summarized: to develop new recombinant collagen types and dosage forms, to explore the mechanism of action of recombinant collagen, and to provide an outlook for the future development and application of recombinant collagen.

In this work,a fluorescent probe PIN was synthesized using indole-3-carbohydrazide and pyrenecarboxaldehyde as raw materials.PIN showed weak fluorescence emission in aqueous solution with acetonitrile volume fraction of 70％.However,when Cu2+was added to this aqueous solution of PIN,a new fluorescence emission peak appeared at 495 nm,and the intensity of this peak gradually increased with the increase of concentration of Cu2+,and also caused a significant change in the fluorescence color of the solution.In contrast,the addition of 15 kinds of other common metal ions did not cause such change.The detection limit of PIN for Cu2+was 78.7 nmol/L,which was much lower than the maximum permitting level of Cu2+in drinking water in hygienic standard for drinking water in China.Therefore,PIN was a highly selective and sensitive fluorescence-enhanced probe for Cu2+.Meanwhile,the addition of Cu2+could also cause a new absorption peak at 440 nm in the ultraviolet-visible absorption spectrum of the aqueous solution of PIN,and meanwhile the colorless PIN solution changed into yellow,exhibiting the performance of PIN as a colorimetric probe for Cu2+.By fitting with the Levenberg-Marquardt algorithm equation,the binding ratio of PIN to Cu2+was 2:1,and the binding constant was 3.42×1012 L2/mol2.In addition,the binding mode of PIN with Cu2+was explored by using proton nuclear magnetic resonance(1H NMR)titration experiments and density functional theory simulations.The results showed that the addition of Cu2+could cause the aggregation of PIN molecules to form excimers,thus showing highly selective recognition.Finally,PIN was made into a simple test strip,which could achieve rapid and convenient fluorescence detection of Cu2+in actual water samples.

Objective:To compare the effects of remimazolam and propofol sedation on pulmonary ventilation in patients undergoing fiberoptic bronchoscopy.Methods:In this randomized controlled trial, 90 American Society of Anesthesiologists Physical Status classification Ⅰ to Ⅲ patients, aged 18-70 yr, with a body mass index of 18-30 kg/m 2, scheduled for elective fiberoptic bronchoscopy and/or treatment under sedation at the Endoscopy Center of the Fourth Affiliated Hospital of Soochow University between November 2021 and December 2022, were divided into 2 groups ( n=45 each) using a random number table: remimazolam group (group R) and propofol group (group P). After intravenous injection of sufentanil 0.1 μg/kg, an initial dose of remimazolam 0.075 mg/kg and propofol 0.9 mg/kg was intravenously injected in group R and group P, respectively. When necessary, additional single doses of remimazolam 0.025 mg/kg or propofol 0.3 mg/kg were administered until the bispectral index value reached 60–80 and the Modified Observer′s Assessment of Alertness/Sedation score was ≤3 (successful sedation). Electrical impedance tomography was conducted immediately after entering the operating room, immediately after successful sedation, immediately after completion of bronchoscopy the procedure, and immediately upon awakening to calculate the tidal impedance variation, center of ventilation, and global inhomogeneity index. The time to successful sedation, emergence time, the lowest SpO 2 during sedation, and occurrence of adverse events such as respiratory depression, injection pain, hypotension, hypertension, tachycardia, bradycardia and dizziness were recorded. Results:Compared with group P, the tidal impedance variation was significantly increased immediately after the procedure, the global inhomogeneity index was decreased immediately after the procedure and upon awakening, the emergence time was prolonged, the lowest SpO 2 was elevated, and the incidence of hypotension and injection pain was decreased in group R ( P<0.05). Conclusions:Compared with propofol sedation, remimazolam sedation has a smaller impact on pulmonary ventilation in patients undergoing fiberoptic bronchoscopy.

[This corrects the article DOI: 10.1016/j.apsb.2022.03.002.].

Antibiotic adjuvants offer a promising strategy for restoring antibiotic sensitivity, expanding antibacterial spectra, and reducing required dosages. Previously, compound 15 was identified as a potential adjuvant for Polymyxin B (PB) against multidrug-resistant (MDR) Pseudomonas aeruginosa DK2; however, its clinical utility was hindered by high cytotoxicity, uncertain in vivo efficacy, and an unclear synergetic mechanism. To address these challenges, we synthesized and evaluated a series of novel benzamide derivatives, with A22 emerging as a particularly promising candidate. A22 demonstrated potent synergistic activity to PB, minimal cytotoxicity, improved water solubility, and broad-spectrum synergism of polymyxins against various clinically isolated MDR Gram-negative strains. In vivo studies using Caenorhabditis elegans and mouse models further confirmed the efficacy of A22. Moreover, A22 effectively suppressed the development of PB resistance in Pseudomonas aeruginosa DK2. Mechanistic investigations revealed that A22 enhances polymyxins activity by inducing reactive oxygen species production, reducing ATP levels, increasing NOX activity, and inhibiting biofilm formation, leading to bacterial death. These findings position A22 as a highly promising candidate for the development of polymyxin adjuvants, offering a robust approach to combating MDR Gram-negative bacterial infections.

The development of Traditional Chinese Medicine(TCM)medical alliances plays a pivotal role in enhancing grassroots TCM service capabilities and meeting public demand for TCM healthcare.However,challenges persist in establishing these alliances,including insufficient Party leadership at primary TCM institutions and deficiencies in clinical services,talent de-velopment,and emergency care capacity.This study examines innovative Party building approaches in public hospitals within the new era context,analyzing practical cases of alliance development.Our findings demonstrate that integrating Party building into the governance structure of medical alliances not only strengthens Party leadership at primary TCM institutions but also significant-ly promotes TCM service development.Systematic analysis of case hospital practices reveals several key insights.Firstly,strengthening top-level design through Party committee leadership is crucial.Secondly,addressing the most pressing public healthcare concerns with genuine commitment forms the foundation.Thirdly,deep integration of Party building with core medical services represents the essential approach.Lastly,policy-responsive innovation based on consolidated achievements serves as the key driver.

AIM:This study aims to investigate the effects of Feixin decoction(FXD)on pyroptosis of pulmo-nary artery smooth muscle cells(PASMCs)by modulating nuclear factor κB(NF-κB)/nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)pathway,and to explore how FXD attenuates hypoxic pulmonary hypertension(HPH)in mice.METHODS:A mouse model of HPH was established using the Sugen 5416 combined hypoxia(SuHx)method.Sixty C57BL/6 mice were randomly divided into 6 groups:control group,SuHx group,sildenafil group,and low-,medium-and high-dose FXD groups,with 10 mice in each group.Five weeks after treatment,echocardiographic pa-rameters,including pulmonary artery acceleration time(PAT),pulmonary artery ejection time(PET),right ventricular anterior wall thickness at diastole(RVAWd)and tricuspid annular plane systolic excursion(TAPSE),were measured.Right ventricular systolic pressure(RVSP)was assessed via right heart catheterization.Right ventricular hypertrophy in-dex(RVHI)was determined by weighing the hearts.Histological examination using HE staining was conducted to observe pathological changes in small pulmonary arteries and the right ventricle,while Masson staining was used to assess fibrosis in the right ventricular wall.Immunofluorescence staining was used to detect co-localized expression of α-smooth muscle actin(α-SMA)with NLRP3,N-terminal fragment of gasdermin D(N-GSDMD)and caspase-1 in the pulmonary arteries.Western blot analysis was conducted to measure the protein levels of NF-κB,p-NF-κB,NLRP3,apoptosis-associated speck-like protein containing a caspase recruitment domain(ASC),N-GSDMD,interleukin(IL)-1β,IL-18 and cleaved caspase-1 in lung tissues.Transmission electron microscopy was employed to observe the ultrastructure of PASMCs.RE-SULTS:Compared with control group,the mice in SuHx group exhibited elevated RVSP and RVHI(P＜0.01),de-creased right heart function(P＜0.01),increased right ventricular wall fibrosis,and pulmonary vascular remodeling(P＜0.01).There was also increased co-localized expression of α-SMA with NLRP3,N-GSDMD and caspase-1 in small pul-monary arteries(P＜0.01),as well as elevated levels of p-NF-κB,NLRP3,ASC,N-GSDMD,IL-1β,IL-18 and cleaved caspase-1 in lung tissues(P＜0.01),indicating induced pyroptosis of PASMCs.Compared with SuHx group,FXD treat-ment significantly reduced RVSP and RVHI,improved right ventricular function,and attenuated right ventricular wall fi-brosis and pulmonary vascular remodeling(P＜0.05 or P＜0.01).Treatment with FXD also decreased the co-localized ex-pression of α-SMA with NLRP3,N-GSDMD and caspase-1 in small pulmonary arteries(P＜0.05 or P＜0.01),and down-regulated the protein expression of p-NF-κB,NLRP3,ASC,N-GSDMD,IL-1β,IL-18 and cleaved caspase-1 in lung tis-sues(P＜0.05 or P＜0.01),thereby attenuating the pyroptosis of PASMCs.CONCLUSION:FXD attenuates pulmonary vascular remodeling and right ventricular dysfunction in a mouse model of HPH.This effect may be attributed to its inhibi-tion of NF-κB/NLRP3 pathway,which subsequently reduces the pyroptosis of PASMCs.

Objective:To explore the impact of group interpersonal psychotherapy (IPT-G) on depressive symptoms and social cognitive function of adolescents with depression.Methods:From June 2021 to December 2023, a total of 75 adolescent inpatients diagnosed with depression were recruited from Hangzhou Seventh People’s Hospital. They were divided into intervention group and control group according to the general information matching principle.Finally, 30 patients in each group completed the study.The patients in control group received antidepressant medication, psychiatric care, and general recreational activities.While the patients in intervention group received IPT-G based on the treatment of control group. Children's depression inventory (CDI), interpersonal trust scale (ITS), self-esteem scale (SES) and the adolescent cognitive style questionnaire (ACSQ) were adopted to assess the depressive symptoms and social cognitive function of adolescent inpatients before and after the intervention and at 3 and 6 months of follow-up, respectively. SPSS 25.0 software was used for statistical analysis. Chi-square test, independent sample t-test and repeated measure ANOVA were used for inter or intra group comparisons. Results:1.Comparison of depression: The time and group interaction effect of CDI scores between the two groups was significant ( F=6.405, P<0.01). Inter group comparison showed that the CDI scores of the intervention group at 3 and 6 months (12.10±5.20, 7.93±2.98) were lower than those of the control group(18.13±7.28, 15.77±5.52) (both P<0.05), and the scores of both groups at the end of the intervention, 3 months after the intervention, and 6 months after the intervention were lower than that before intervention (all P<0.05).2.Comparison of social cognitive function: (1)There were significant time and group interaction effect of ITS score and SES score( F=5.871, 6.594, both P<0.01).Inter group comparison showed that the ITS score(78.97±7.63, 83.03±7.42) and the SES score(28.00±4.00, 30.30±3.21) of intervention group at 3 months and 6 months after intervention were all higher than those of control group(ITS: (71.70±12.29, 73.90±12.79); SES: (24.37±5.08, 25.80±4.10)(all P<0.05).Intra group comparison showed that the ITS and the SES scores of the intervention group at 3 months and 6 months after intervention were all higher than before intervention( P<0.05).The ITS score of the control group at 6 months after intervention was higher than before intervention( P<0.05).The SES scores of the control group at three time points after intervention were all higher than those before intervention(all P<0.05).(2)Comparison of ACSQ scores: There were significant time and group interaction effects for the scores of academic stability, academic specificity, interpersonal introversion and extroversion, and interpersonal stability, interpersonal specificity( F=5.414, 9.294, 3.440, 8.231, 10.669, all P<0.05). Inter group comparison showed that the academic stability, academic specificity, interpersonal stability, and interpersonal specificity scores of the intervention group were lower than those of the control group at 3 and 6 months after intervention (all P<0.05).Intra group comparison showed that the academic stability, academic specificity, interpersonal introversion and extroversion, interpersonal stability, and interpersonal specificity scores of the intervention group at 3 months and 6 months after the intervention were lower than those before the intervention (all P<0.05).After 3 and 6 months of intervention, the academic stability, academic specificity, interpersonal introversion and extroversion, interpersonal stability, and interpersonal specificity scores of the control group were all lower than those before intervention (all P<0.05). Conclusion:IPT-G can not only alleviate depressive symtoms, but also improve social cognitive function.