Organoids have become an important technological platform in cancer research,but simulating the primary tumor tissue structure and function still presents problems.The development of gene-editing technology,especially when combined with tumor organoids,provides a new approach for accurately and comprehensively simulating the in vivo characteristics of tumor models.Introducing specific gene mutations or correcting mutations in tumor organoids through gene-editing technology can allow detailed analysis of the mechanisms of tumor initiation and progression,as well as exploring potential therapeutic targets,accelerating the drug-screening process,and providing new insights for personalized cancer treatment.This article reviews the formation of tumor organoids and the technical aspects of gene-editing strategies,emphasizing their unique applications and prospects in tumor organoids.We also propose that accurately simulating the in vivo microenvironment,promoting the standardization and stability of organoid gene-editing technology,and optimizing the efficiency of gene editing can accelerate the application of organoids in precision medicine research.

Organoids have become an important technological platform in cancer research,but simulating the primary tumor tissue structure and function still presents problems.The development of gene-editing technology,especially when combined with tumor organoids,provides a new approach for accurately and comprehensively simulating the in vivo characteristics of tumor models.Introducing specific gene mutations or correcting mutations in tumor organoids through gene-editing technology can allow detailed analysis of the mechanisms of tumor initiation and progression,as well as exploring potential therapeutic targets,accelerating the drug-screening process,and providing new insights for personalized cancer treatment.This article reviews the formation of tumor organoids and the technical aspects of gene-editing strategies,emphasizing their unique applications and prospects in tumor organoids.We also propose that accurately simulating the in vivo microenvironment,promoting the standardization and stability of organoid gene-editing technology,and optimizing the efficiency of gene editing can accelerate the application of organoids in precision medicine research.

Pancreatic ductal adenocarcinoma(PDAC)is a common type of pancreatic cancer that is insidious,develops rapidly,and is highly malignant.Traditional treatment strategies are ineffective for PDAC because of its rich extracellular matrix(ECM).Cancer-associated fibroblast(CAF)are the most important component of the ECM,and interact with other immune components in the tumor microenvironment(TME)by secreting numerous effector molecules to form an immunosuppressive TME,which may then allow cancer cells to evade immune system surveillance,promote tumor growth,invasion,and metastasis,and induce ECM remodeling and drug resistance.This review summarizes research progress on the application of targeted CAF in PDAC immunotherapy.We focus on exploring research strategies that promote the transition of TME from an immunosuppressive to an immune-activated state through depleting CAF,inhibiting effector molecules secreted by CAF,reprogramming CAF,and limiting CAF-induced ECM remodeling.This review aims to support the production of more effective therapeutic strategies and provide new method for the immunotherapy of PDAC.

Pancreatic ductal adenocarcinoma(PDAC)is a common type of pancreatic cancer that is insidious,develops rapidly,and is highly malignant.Traditional treatment strategies are ineffective for PDAC because of its rich extracellular matrix(ECM).Cancer-associated fibroblast(CAF)are the most important component of the ECM,and interact with other immune components in the tumor microenvironment(TME)by secreting numerous effector molecules to form an immunosuppressive TME,which may then allow cancer cells to evade immune system surveillance,promote tumor growth,invasion,and metastasis,and induce ECM remodeling and drug resistance.This review summarizes research progress on the application of targeted CAF in PDAC immunotherapy.We focus on exploring research strategies that promote the transition of TME from an immunosuppressive to an immune-activated state through depleting CAF,inhibiting effector molecules secreted by CAF,reprogramming CAF,and limiting CAF-induced ECM remodeling.This review aims to support the production of more effective therapeutic strategies and provide new method for the immunotherapy of PDAC.

Objective:To summarize the clinical and genetic features of 7 patients with a mild form of Geleophysic dysplasia type 2(GD2)/Acromicric dysplasia(AD) induced by fibrillin 1(FBN1) gene mutation from one Chinese family.Methods:A Chinese pedigree of mild GD2/AD treated at the Pediatric Endocrinology Department at the First Affiliated Hospital of Sun Yat-sen University between August 2017 and May 2022 was collected. Whole-exome genetic sequencing of the FBN1 gene were performed to establish the diagnosis. Additionally, a literature review was further conducted.Results:In this family, among 13 individuals spanning three generations, there were 7 affected cases, including 1 adult female, 1 adult male, and 5 children. All individuals exhibited postnatal growth failure, severe disproportionate short stature, and lacked typical facial features. Exome sequencing and Sanger sequencing confirmed the presence of a heterozygous missense mutation c. 5099A>G(p.Tyr1700Cys) in exon 42 of the FBNI gene in 6 affected individuals(Ⅱ-1, Ⅲ-1 to Ⅲ-5), which was identified as a pathogenic mutation. This mutation was previously reported in a Chinese classical achondroplasia(AD) family. Based on comprehensive genetic analysis, clinical features, and multisystem evaluation, 3 cases were diagnosed with mild type 2 growth hormone deficiency(GD2), and 4 cases were diagnosed with mild AD. Recombinant human growth hormone(rhGH; 1.1-1.4 IU·kg -1·week -1) was applied to all the 5 children, and additional gonadotropin releasing hormone analogue(GnRHa) was administered to the 2 girls in late puberty, resulting in certain growth-promoting effect. Conclusions:The c. 5099A>G(p.Tyr1700Cys) mutation not only leads to the classical type of achondroplasia(AD) as reported in the literature but also causes the non-classical GD2 or AD(mild GD2/AD). Further research is warranted to investigate the long-term therapeutic effects of rhGH treatment.

Background The health effects of forest therapy have been widely recognized, while the previous studies mostly focused on a single activity mode of forest walks. The effects of different types of forest therapy activities remain unclear. Objective To explore the effects of short-term forest therapy on cardiopulmonary health, psychological health, and sleep quality, and the health effects of different types of forest therapy activities, aiming to provide population empirical study data for the development of forest therapy. Methods A self-control study was conducted in a national forest park in suburb of Beijing from August to September 2018. A total of 31 healthy college students were recruited as the study subjects, with a total forest stay for 3 days and 2 nights. During the period of study, each subject practiced walking therapy, sitting therapy with five senses experience (sitting therapy thereafter), and handmade work therapy, successively. Each type of forest therapy lasted about 2 h. Changes of blood pressure, oxygen saturation (SpO₂), lung function, and fractional exhaled nitric oxide (FeNO) were estimated by measuring corresponding indicators before and after the forest therapy. Psychological health and sleep quality were assessed by Profile of Mood States and Pittsburgh Sleep Quality Index respectively at the same time. Mixed effects models were used to analyze the changes of these health indicators. The health effects of different types of forest therapy activities were further analyzed. Results The average age and body mass index of subjects in this study were (24.5±2.6) years and (20.7±1.7) kg·m⁻², respectively. After a short-term forest therapy, the selected indicators of cardiopulmonary health, psychological health, and sleep quality of subjects were all improved. In particular, the pulse pressure (PP) and FeNO decreased by 3.02 mmHg and 1.10 ppb, respectively, while the SpO₂ and peak expiratory flow (PEF) increased by 0.65% and 0.50 L·s⁻¹, respectively, and the negative emotion and global sleep quality also presented significant positive changes (all P<0.05). Furthermore, different therapy activities presented differential effects in the health indicators. Walking therapy significantly improved pulmonary function, SpO₂, and confusion (CON) emotion, in which the SpO₂, forced expiratory volume in the first second (FEV₁), and forced vital capacity (FVC) increased by 0.48%, 0.14 L, and 0.12 L, respectively, and the score of CON decreased by 0.97 (all P<0.05). Sitting therapy significantly reduced blood pressure and tension (TEN) emotion of subjects, including a decrease of the systolic blood pressure (4.45 mmHg), PP (4.19 mmHg), and the score of TEN (0.84) (all P<0.05). The diastolic blood pressure (DBP) increased slightly after handmade work therapy (ΔDBP=2.44 mmHg, P=0.016), but there were no significant changes in other indicators. Conclusion Short-term forest therapy could significantly improve cardiopulmonary health, psychological health, and sleep quality of young healthy individuals, and different types of forest therapy activities may have differential health effects.

Objective To explore the effect of lung strain on breathing mechanics in dogs with acute respiratory distress syndrome (ARDS).Methods Twenty-four healthy male Beagle dogs were recruited to reproduce medium ARDS models with venous injection of 0.18 mL/kg oleic acid, and they were randomly assigned to five groups with random numbers table method. In lung protective ventilation (LPV) group (n = 4), the ventilation was supported for 24 hours with tidal volume (VT) at 6-8 mL/kg, and in lung strain 1.0, 1.5, 2.0, 2.5 groups (S1.0, S1.5, S2.0, S2.5 groups), the VT was calculated from lung strain, the volume recruitment by positive end expiratory pressure (VPEEP) and functional residual capacity (FRC). Five groups were given mechanical ventilation for 24 hours or until reaching the end point of the experiment [when the dosage of norepinephrine was higher than 1.4μg·kg-1·min-1, the blood pressure was still lower than 60 mmHg (1 mmHg = 0.133 kPa) for more than 30 minutes, which was regarded as the end point of the experiment]. Static lung compliance (Cst), airway plateau pressure (Pplat) and lung stress during the experiment were recorded. Linear regression analysis was used to fit the regression equations of lung strain and Cst descending rate,Pplat and lung stress for analyzing their relationships.Results The VT of group LPV was (7.1±0.5) mL/kg. With the increase of lung strain, VT was gradually increased. VT of group S1.0 [(7.3±1.8) mL/kg] was similar to group LPV. VT of groups S1.5, S2.0, S2.5 was significantly higher than that of group LPV (mL/kg: 13.3±5.5, 18.7±5.4, 20.1±7.4 vs. 7.1±0.5, allP < 0.05). Moreover, under the same lung strain, the difference in VT among individuals was large. The Cst of each group was decreased significantly at the end of the experiment as compared with that before model reproduction. With the increase of lung strain, the rate of Cst descending was increased, Cst dropped more significantly in groups S2.0 and S2.5 than that in groups S1.0 and S1.5 [(48.0±15.0)％, (54.4±9.5)％ vs. (25.9±13.7)％, (38.6±8.1)％, all P < 0.05]. Pplat and pulmonary stress at model reproduction in all groups were significantly higher than those before model reproduction, and they increased with the prolongation of ventilation time. Pplat and lung stress at 4 hours of ventilation in group S1.5 were significantly higher than those in group LPV [Pplat (cmH2O, 1 cmH2O = 0.098 kPa):26.2±2.3 vs. 20.2±4.2, lung stress (cmH2O): 20.5±2.0 vs. 16.6±2.5, bothP < 0.05], and they increased with lung strain increasing till to the end of experiment. It was shown by correlation analysis that lung strain was positively related with Cst descending rate, Pplat and lung stress at 4 hours of ventilation (rvalue was 0.716, 0.660, 0.539, respectively, allP < 0.05), which indicated a strong linear correlation. It was shown by fitting linear regression analysis that when lung strain increased by 1, Cst descending rate increased by 19.0％ [95％ confidence interval (95％CI) = 14.6-23.3, P = 0.000], Pplat increased by 10.8 cmH2O (95％CI = 7.9-13.7,P = 0.002), and the lung stress increased by 7.4 cmH2O (95％CI = 4.7-10.2,P = 0.002).Conclusion In animal ARDS models, the larger the lung strain, the higher the Pplat and lung stress during mechanical ventilation, VT originated for lung strain 2.0 and 2.5 may further reduce Cst in ARDS models, when lung strain over 1.5, Pplat and lung stress increased significantly, which exceeded the safe range of LPV (35 cmH2O and 25 cmH2O, respectively), and further aggravated ventilator induced lung injury (VILI).

Objective To study the effect of TLR4 activation with LPS on BMP9-induced osteogenic differentiation of immortalized mouse embryonic fibroblasts ( iMEFs).Methods The activation of TLR4/NF-κB signaling path-way was detected by ICC.iMEFs were treated with LPS,BAY11-7082,Adnovirus GFP and BMP9.The early osteo-genic differentiation capability of iMEFs was detected by ALP staining and quantitative assay .The later osteogenic differentiation capability was detected by alizarin red S staining .The expression of later osteogenic differentiation marker gene OCN and OPN were detected by PCR and Western blot .The change of p-Smad1/5/8 was detected by Western blot.The expression of Runx2 and Dlx5 were detected by PCR and Western blot .Results LPS can effec-tively stimulate TLR4/NF-κB signaling pathway .TLR4 activation inhibited BMP 9-induced osteogenic differentiation . BMP9-induced osteogenic differentiation related gene and Smad 1/5/8 signaling activation were inhibited by TLR4 activation .The inhibition effect was partly reversed by BAY 11-7082 ( P<0.05 ) .Conclusions TLR4 activation with LPS can inhibit BMP9-induced osteogenic differentiation of iMEFs cells via NF-κB signaling pathway .

AIM:To investigate the effect of bone morphogenetic proteins 9 (BMP9) on the migration and in-vasion abilities of human lung squamous-cell carcinoma NCI-H520 cells and its mechanism.METHODS:The expression of BMP9 at mRNA and protein levels in the NCI-H520 cells and human bronchial epithelial ( HBE) cells was detected by RT-PCR and Western blot.The NCI-H520 cells were transfected with the recombinant adenovirus AdBMP9 and the expres-sion of BMP9 at mRNA and protein levels was validated by RT-PCR and Western blot.The migration and invasion abilities of the NCI-H520 cells were determined by wound-healing and Transwell assays.The mRNA and protein levels of the migra-tion-related factor matrix metalloproteinase 2 (MMP2) were detected by RT-PCR and Western blot.The level of phospho-rylated Smad1/5 (p-Smad1/5) was detected by Western blot.Meanwhile, NCI-H520 cells were treated with BMP specific antagonist AdNoggin and AdBMP9.The level of p-Smad1/5 and the cell migration ability were measured by Western blot, wound-healing and Transwell assays.RESULTS:The expression of BMP9 at mRNA and protein levels was lower in NCI-H520 cells than that in HBE cells.After AdBMP9 was stably transfected into the NCI-H520 cells, the expression of BMP9 at mRNA and protein levels was significantly up-regulated, cell migration and invasion abilities were significantly de-creased, and the mRNA and protein levels of MMP2 were decreased.Meanwhile, the level of p-Smad1/5 was increased. Noggin reversed BMP9-caused the increase in p-Smad1/5 and the decrease in cell migration ability.CONCLUSION:O-ver-expression of BMP9 inhibits the migration and invasion abilities of lung squamous-cell carcinoma NCI-H520 cells.The activation of BMP-Smad signaling pathway may be involved in this inhibitory process.