Immunotherapy has emerged as a revolutionary approach to treat immune-related diseases. Dendritic cells (DCs) play a pivotal role in orchestrating immune responses, making them an attractive target for immunotherapeutic interventions. Modulation of gene expression in DCs using genome editing techniques, such as the CRISPR-Cas system, is important for regulating DC functions. However, the precise delivery of CRISPR-based therapies to DCs has posed a significant challenge. While lipid nanoparticles (LNPs) have been extensively studied for gene editing in tumor cells, their potential application in DCs has remained relatively unexplored. This study investigates the important role of cholesterol in regulating the efficiency of BAMEA-O16B lipid-assisted nanoparticles (BLANs) as carriers of CRISPR/Cas9 for gene editing in DCs. Remarkably, BLANs with low cholesterol density exhibit exceptional mRNA uptake, improved endosomal escape, and efficient single-guide RNA release capabilities. Administration of BLAN_mCas9/gPD-L1 results in substantial PD-L1 gene knockout in conventional dendritic cells (cDCs), accompanied by heightened cDC1 activation, T cell stimulation, and significant suppression of tumor growth. The study underscores the pivotal role of cholesterol density within LNPs, revealing potent influence on gene editing efficacy within DCs. This strategy holds immense promise for the field of cancer immunotherapy, offering a novel avenue for treating immune-related diseases.

Tricellulin, a key tricellular tight junction (TJ) protein, is essential for maintaining the barrier integrity of acinar epithelia against macromolecular passage in salivary glands. This study aims to explore the role and regulatory mechanism of tricellulin in the development of salivary gland hypofunction in Sjögren's syndrome (SS). Employing a multifaceted approach involving patient biopsies, non-obese diabetic (NOD) mice as a SS model, salivary gland acinar cell-specific tricellulin conditional knockout (Tric^CKO) mice, and IFN-γ-stimulated salivary gland epithelial cells, we investigated the role of tricellulin in SS-related hyposalivation. Our data revealed diminished levels of tricellulin in salivary glands of SS patients. Similarly, NOD mice displayed a reduction in tricellulin expression from the onset of the disease, concomitant with hyposecretion and an increase in salivary albumin content. Consistent with these findings, Tric^CKO mice exhibited both hyposecretion and leakage of macromolecular tracers when compared to control animals. Mechanistically, the JAK/STAT1/miR-145 axis was identified as mediating the IFN-γ-induced downregulation of tricellulin. Treatment with AT1001, a TJ sealer, ameliorated epithelial barrier dysfunction, restored tricellulin expression, and consequently alleviated hyposalivation in NOD mice. Importantly, treatment with miR-145 antagomir to specifically recover the expression of tricellulin in NOD mice significantly alleviated hyposalivation and macromolecular leakage. Collectively, we identified that tricellulin deficiency in salivary glands contributed to hyposalivation in SS. Our findings highlight tricellulin as a potential therapeutic target for hyposecretion, particularly in the context of reinforcing epithelial barrier function through preventing leakage of macromolecules in salivary glands.
Sjogren's Syndrome/complications* ; Animals ; Xerostomia/etiology* ; Mice ; Mice, Inbred NOD ; MARVEL Domain Containing 2 Protein/metabolism* ; Humans ; Mice, Knockout ; Disease Models, Animal ; Interferon-gamma ; Salivary Glands/metabolism* ; Tight Junctions/metabolism* ; MicroRNAs/metabolism* ; Female

Objective China is among the 30 countries with a high burden of tuberculosis(TB)worldwide,and TB remains a public health concern.Kashgar Prefecture in the southern Xinjiang Autonomous Region is considered as one of the highest TB burden regions in China.However,molecular epidemiological studies of Kashgar are lacking. Methods A population-based retrospective study was conducted using whole-genome sequencing(WGS)to determine the characteristics of drug resistance and the transmission patterns. Results A total of 1,668 isolates collected in 2020 were classified into lineages 2(46.0%),3(27.5%),and 4(26.5%).The drug resistance rates revealed by WGS showed that the top three drugs in terms of the resistance rate were isoniazid(7.4%,124/1,668),streptomycin(6.0%,100/1,668),and rifampicin(3.3%,55/1,668).The rate of rifampicin resistance was 1.8%(23/1,290)in the new cases and 9.4%(32/340)in the previously treated cases.Known resistance mutations were detected more frequently in lineage 2 strains than in lineage 3 or 4 strains,respectively:18.6%vs.8.7 or 9%,P<0.001.The estimated proportion of recent transmissions was 25.9%(432/1,668).Multivariate logistic analyses indicated that sex,age,occupation,lineage,and drug resistance were the risk factors for recent transmission.Despite the low rate of drug resistance,drug-resistant strains had a higher risk of recent transmission than the susceptible strains(adjusted odds ratio,1.414;95%CI,1.023-1.954;P = 0.036).Among all patients with drug-resistant tuberculosis(DR-TB),78.4%(171/218)were attributed to the transmission of DR-TB strains. Conclusion Our results suggest that drug-resistant strains are more transmissible than susceptible strains and that transmission is the major driving force of the current DR-TB epidemic in Kashgar.

OBJECTIVE: To examine the association of handgrip strength with aortic stenosis incidence among adults aged 60 years and older. METHODS: We conducted a cohort study using the UK Biobank data to assess the relationship between handgrip strength and incident aortic stenosis in individuals aged 60 years and older. Handgrip strength was measured using a Jamar J00105 hydraulic hand dynamometer. Adjusted Cox proportional hazards regression models were conducted to assess the association between handgrip strength and incident aortic stenosis. RESULTS: We included 157,097 UK Biobank participants (78,151 women and 78,946 men) in our study, with mean age of 64 ± 2.9 years. During a median follow-up of 8.1 (7.4-8.8) years, 1543 (1.0%) participants developed incident aortic stenosis. Compared with those with the lowest handgrip strength (tertile 1), the adjusted hazard ratios (95% confidence interval) of incident aortic stenosis in the middle (tertile 2) and the highest (tertile 3) were 0.86 (0.77-0.97) and 0.76 (0.67-0.87), respectively. CONCLUSIONS Higher handgrip strength was associated with lower risk of developing aortic stenosis in older adults. Future studies warrant preventive strategies for older adults with lower handgrip strength.

Aim To explore the potential mechanism of action of Codonopsis Poria in the treatment of alco-holic liver disease(ALD).Methods TCMSP and Swiss Target Prediction were used to obtain the active ingredients and targets of Codonopsis Poria;OMMI,DisGeNET and GeneCards databases were used to obtain the targets of ALD;STRING database was used to construct the PPI network;and Bioconductor soft-ware was used to analyze the enrichment of GO and KEGG pathways.Cytoscape 3.7.1 software was used to construct the drug-component-target-disease network of Codonopsis Poria for ALD treatment,and key targets were screened for molecular docking;the effects of Codonopsis Poria on ALD rats were verified by experi-ments.Results The removal of duplicate targets ob-tained 36 chemical components and 529 potential ac-tion targets.GO enrichment analysis:2 245 biological processes,74 cellular components,125 molecular functions.KEGG enrichment analysis:159 signaling pathways,mainly involving PI3K-Akt,MAPK,AGE-RAGE signaling pathways.Molecular docking showed that AKT1,MMP9 and other targets may be the key targets of Codonopsis Poria in the treatment of ALD.Experiments showed that Codonopsis Poria could im-prove the inflammation level of hepatocytes in ALD rats and reduce the levels of TC,TG,AST,ALT and GGT in ALD rats,PCR assay concluded that Codonopsis Po-ria could reduce the expression of PI3 K and AKT,and electron microscopy results showed that Codonopsis Po-ria could affect the autophagy of cells.Conclusions It is initially revealed that Codonopsis Poria may atten-uate inflammatory cell infiltration by affecting the ex-pression of AKT,TNF and MAPK,and it is hypothe-sized that Codonopsis Poria may affect autophagy through the PI3K-Akt signaling pathway,thus treating ALD,which is initially verified by PCR assay to pro-vide a basis for in-depth explanation of the molecular mechanism of Codonopsis Poria medicinal pairs in the treatment of ALD.

Objective To investigate the effect of aspirin on oxygen glucose deprivation/reoxygen-ation(OGD/R)-induced injury in mouse hippocampal neuron HT22 cells by regulating ferropto-sis.Methods HT22 cells were randomly divided into control group,model group,low-,medium-and high-dose groups(n=3).Cellular OGD/R injury model was established in the other 4 groups except the control group.Aspirin of 100,200 and 400 μg/ml was used to treat the cells from the above 3 treatment groups,respectively.Cell viability was detected by CCK-8 assay.The contents of TNF-α,IL-1β and IL-6 were detected by ELISA.The contents of SOD,catalase,glutathione,reac-tive oxygen species(ROS),lactate dehydrogenase(LDH),Fe2+and MDA were detected by the corresponding reagent kits.Western blot analysis was performed to determine the expression of solute carrier family 7 members 11(SLC7A11),glutathione peroxidase 4(GPX4)and Acyl-CoA synthase long chain member 4(ACSL4).Results The model group had significantly lower cell vi-ability than the control group(0.49±0.07 vs 1.00±0.12,P＜0.01),but the viability of the low-,medium-and high-dose groups were higher than that of the model group(0.72±0.10 vs 0.49±0.07,P＜0.05;0.87±0.10 vs 0.49±0.07,P＜0.01;0.93±0.07 vs 0.49±0.07,P＜0.01).Compared with the control group,the contents of TNF-α,IL-1β,IL-6,ROS,LDH,Fe2 and MDA and the protein expression of ACSL4 were significantly increased,while the contents of SOD,catalase,glutathione and protein levels of SLC7A11 and GPX4 were obviously decreased in the model group(P＜0.01).Compared with model group,aspirin treatment reversed all above indicators no matter its doses(P＜0.05,P＜0.01).Conclusion Aspirin alleviates OGD/R-induced neuronal in-jury through regulating ferroptosis in mouse neuron HT22 cells in a dose-dependent manner.

BACKGROUND: Pneumonia-like primary pulmonary lymphoma (PPL) was commonly misdiagnosed as infectious pneumonia, leading to delayed treatment. The purpose of this study was to establish a computed tomography (CT)-based radiomics model to differentiate pneumonia-like PPL from infectious pneumonia. METHODS: In this retrospective study, 79 patients with pneumonia-like PPL and 176 patients with infectious pneumonia from 12 medical centers were enrolled. Patients from center 1 to center 7 were assigned to the training or validation cohort, and the remaining patients from other centers were used as the external test cohort. Radiomics features were extracted from CT images. A three-step procedure was applied for radiomics feature selection and radiomics signature building, including the inter- and intra-class correlation coefficients (ICCs), a one-way analysis of variance (ANOVA), and least absolute shrinkage and selection operator (LASSO). Univariate and multivariate analyses were used to identify the significant clinicoradiological variables and construct a clinical factor model. Two radiologists reviewed the CT images for the external test set. Performance of the radiomics model, clinical factor model, and each radiologist were assessed by receiver operating characteristic, and area under the curve (AUC) was compared. RESULTS: A total of 144 patients (44 with pneumonia-like PPL and 100 infectious pneumonia) were in the training cohort, 38 patients (12 with pneumonia-like PPL and 26 infectious pneumonia) were in the validation cohort, and 73 patients (23 with pneumonia-like PPL and 50 infectious pneumonia) were in the external test cohort. Twenty-three radiomics features were selected to build the radiomics model, which yielded AUCs of 0.95 (95% confidence interval [CI]: 0.94-0.99), 0.93 (95% CI: 0.85-0.98), and 0.94 (95% CI: 0.87-0.99) in the training, validation, and external test cohort, respectively. The AUCs for the two readers and clinical factor model were 0.74 (95% CI: 0.63-0.83), 0.72 (95% CI: 0.62-0.82), and 0.73 (95% CI: 0.62-0.84) in the external test cohort, respectively. The radiomics model outperformed both the readers' interpretation and clinical factor model ( P <0.05). CONCLUSIONS The CT-based radiomics model may provide an effective and non-invasive tool to differentiate pneumonia-like PPL from infectious pneumonia, which might provide assistance for clinicians in tailoring precise therapy.
Humans ; Retrospective Studies ; Pneumonia/diagnostic imaging* ; Analysis of Variance ; Tomography, X-Ray Computed ; Lymphoma/diagnostic imaging*

Objective: : To investigate the efficacy and safety of the posterior endoscopic cervical foraminotomy (PECF) using ultrasonic osteotome for the treatment of cervical osseous foraminal stenosis，focusing on introduction of the advantages of ultrasonic osteotome in partial pediculectomy and ventral osteophyte resection in PECF. Methods: : Nineteen patients with cervical osseous foraminal stenosis who underwent PECF using ultrasonic osteotome in our institution between April 2018 and April 2021 were enrolled in this study. All the patients were followed up more than 12 months. The patients’ medical data, as well as pre- and postoperative radiologic findings were thoroughly investigated. The visual analogue score (VAS), Japanese Orthopaedic Association (JOA) score, cervical dysfunction index (Neck disability index, NDI), and modified MacNab criteria were used to assess the surgical efficacy. Results: : All the patients were successfully treated with PECF using ultrasonic osteotome. The pre- and postoperative VAS, NDI, and JOA scores were significantly improved (p<0.05). According to the modified MacNab criteria, 17 patients were assessed as “excellent”, two patients were assessed as “good” at the last follow-up. There was no dura tear, nerve root damage, incision infection, neck deformity, or other complications. Conclusion : Adequate nerve root decompression can be accomplished successfully with the help of ultrasonic osteotome in PECF, which has the advantage of reducing the probability of damage to the nerve root and dura mater, in addition to the original merits of endoscopic surgery.

To explore the predictive value of preoperative serum CYFRA 21-1 in colorectal cancer (CRC) resection patients. In this retrospective study, 456 patients with CRC who received surgical treatment in the Department of General Surgery, Affiliated Hospital of Nantong University from January 2016 to February 2018 were analyzed. Preoperative CYFRA 21-1, CEA, CA19-9 and pathological data of the study subjects were collected. Determine the cut-off value of CYFRA 21-1 based on the X-tile. Chi-square test or Fisher exact probability test were used to compare clinicopathological features in different CYFRA 21-1 level groups. Univariate and multivariate regression analysis of factors affecting 5-year overall survival (OS) and disease-free survival (DFS). Kaplan-Meier survival curves were used to analyze 5-year differences in OS and DFS in CRC patients with different levels of CYFRA 21-1, CEA and CA19-9. Receiver operating characteristic(ROC) was adopted. ROC curves were used to analyze the prognostic efficacy of CYFRA21-1 for CRC, and nomogram maps were used to predict 1, 3, and 5-year survival rates. The results showed that the optimal cut-off values of serum CYFRA 21-1, CEA and CA19-9 were 4.9 ng/ml, 29.2 ng/ml and 72.8 U/ml, respectively. Different gender, tumor size, location, degree of differentiation, depth of invasion, lymph node metastasis and tumor node metastasis (TNM) classification stage were significantly different between the two groups with high and low CYFRA 21-1, the P-values were 0.018,<0.001,<0.001,<0.001, 0.002, 0.001, 0.003, respectively. CYFRA 21-1 (≥4.9 ng/ml) was an independent risk factor for 5-year OS (HR: 4.008, 95%CI: 2.309-6.958, P<0.001) and DFS (HR: 3.75, 95%CI: 2.227-6.314, P<0.001) in CRC patients. CYFRA 21-1 predicts a 5-year AUC of 0.725 and 0.720 for OS and DFS, respectively, and 0.804 and 0.827 for the combination of CEA and CA19-9. Based on the results of multivariate Cox regression analysis, nomogram graphs of OS and DFS were established, the C-indexes were 0.799 and 0.803, respectively. In conclusion, preoperative serum CYFRA 21-1 level may be an independent risk factor affecting the prognosis of patients with colorectal cancer. The prognostic model established by CYFRA 21-1 combined with CEA, CA19-9 and TNM stages may provide references for the prevention of CRC recurrence and clinical decision-making.
Humans ; Colorectal Neoplasms/pathology* ; CA-19-9 Antigen ; Retrospective Studies ; Neoplasm Staging ; Neoplasm Recurrence, Local/pathology* ; Biomarkers, Tumor

ObjectiveTo study the effect of Bazi Bushen capsules on delaying the aging process of naturally aging mice and its mechanism. MethodThe mice were randomly divided into four groups according to their body weight， namely， aging group， low-dose Bazi Bushen capsules group （1 g·kg^-1）， high-dose Bazi Bushen capsules group （2 g·kg^-1）， and rapamycin group （0.002 g·kg^-1）. The debilitating signs were detected by behavioral tests and the weakness index was measured. The percentages of spleen T and B lymphocytes， effector T cells （TE）， memory T cells （TM）， naive T cells （TN）， helper T cells （Th）， cytotoxic T cells （Tc） ，Th1 cells， Th2 cells， and regulatory T cells （Treg） were determined by flow cytometry. Cell proliferation and the cell counting kit-8 （CCK-8） assay was used to detect the proliferation of lymphocytes in mice. The hematoxylin-eosin （HE） staining was used to observe the histopathological changes in the mouse spleen. The expression of cyclin-dependent kinase inhibitor 2A （p16） and cyclin-dependent kinase inhibitor 1A （p21） was detected by immunohistochemistry （IHC）. The mRNA expression of senescence-related proteins p16 and p21 was determined by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）， and the inflammatory cytokines， including tumor necrosis factor-α （TNF-α）， interferon-γ （IFN-γ）， interleukin （IL）-1β， IL-2， IL-4， IL-10， and IL-12 p70， in peripheral blood of mice were detected by Luminex. ResultAs compared with the aging group， mice in the Bazi Bushen capsules and rapamycin groups showed significantly improved debilitating signs and reduced weakness index scores （P<0.05， P<0.01）， increased proportions of T cells， TN cells， Tc cells， Th2 cells， and Treg cells in the spleen， decreased proportions of TE cells， TM cells， Th cells， Th1 cells （P<0.05， P<0.01）， and increased proliferation of splenic lymphocytes （P<0.05， P<0.01）. In the Bazi Bushen capsules and rapamycin groups， clear structure of the red and white marrow marginal zone was observed in the spleen of mice， the area of the white marrow was increased， and the area of the red marrow was correspondingly decreased. The protein and mRNA expression of aging-related proteins p16 and p21 in the spleen was decreased （P<0.01）， the levels of serum pro-inflammatory cytokines IL-1β， IL-2， IL-12 p70， IFN-γ， and TNF-α levels were decreased， and the levels of IL-4 and IL-10 were increased （P<0.05， P<0.01） in the Bazi Bushen capsules and rapamycin groups as compared with the aging group. ConclusionBazi Bushen capsules have the effect of regulating the debilitating signs of natural aging mice， regulating the immune homeostasis and inflammation level of the body， and reducing cell aging.