Objective To analyze the composition and function of residual cells in pre-transplantation human thymic slices by single-cell transcriptomics sequencing(scRNA-seq),and established a quality assessment method for thymic slices based on the expression levels of molecular markers in the culture supernatant.Methods The discarded thymus from 18 patients with congenital heart disease undergoing surgical treatment in Department of Cardiothoracic Surgery of Children's Hospital Affiliated to Chongqing Medical University from May 2023 to January 2024 were collected and prepared into thymic slices.After the slices were cultured in vitro for 14 d,scRNA-seq was employed to identify the residual cell types,and gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)enrichment analysis was performed to analyze the biological function of the residual cells.Then based on the literature concerning thymic slice culture,the molecular markers indicating thymocyte function were screened out.ELISA was applied to detect the changes in protein levels of molecular markers in the supernatant.Receiver operating characteristic(ROC)curve was plotted and assess the value of the molecular markers in the supernatant in evaluating the quality of thymic slices with area under the curve(AUC).Then,the qualified and unqualified thymic slices determined by our obtained molecular markers were transplanted subcutaneously into male nude mice(6～8 weeks old,weighing 14～17 g),respectively,and the male nude mice without transplantation of the thymic slices served as control group.Flow cytometry and histologic analysis were utilized to observe the immune reconstitution after transplantation.Results ① scRNA-seq identified 11 cell types in thymic slices,dominated with epithelial cells,fibroblasts,and T cells.GO and KEGG enrichment analysis showed that epithelial cells were involved in enrichment entries related to chemotaxis,epithelial cell development,cell matrix adhesion and tight junction;fibroblasts were involved in enrichment entries related to extracellular matrix,epithelial cell proliferation,negative regulation of cell migration,and regulation of actin cytoskeleton;T cells were mainly related to T cell differentiation,regulation of T cell activation,T cell apoptosis,and T cell receptor signaling.② Molecular markers,CCL19,CCL21,CXCL12,CXCL16,IL16 and SELL were identified to indicate thymocyte function.Compared with the levels of the first day,the protein secretions of CCL19,CCL21,CXCL12 and CXCL16 were significantly increased during in vitro culture(P<0.05),while the protein secretions of IL16 and L-selectin(protein form of SELL)were significantly decreased(P<0.05).The combined predictor Pre1 from subset of cytokines(IL16 and L-selectin)had the highest value in the quality assessment of thymic slices after 1 d of culture(AUC=0.883),and the combined predictor Pre2 from subset of cytokines(CCL19,CCL21,CXCL12 and CXCL16)had the highest value in the quality assessment after 14 d of culture(AUC=0.948).③ Transplantation in nude mice indicated that the qualified thymic slices could develop to thymus structure in vivo,and effectively increase the proportion of T cells in peripheral blood(P<0.01),while the unqualified thymic slices could not obtain the reconstitution of T cell development.Conclusion The main residual component cells in thymic slices are epithelial cells,fibroblasts and T cells.IL16 and L-selectin can be used as potential indicators to determine the quality of donor thymic samples.CCL19,CCL21,CXCL12 and CXCL16 can effectively evaluate the quality of thymic slices before transplantation.

Echocardiography is an important imaging technique in mithal valve transcatheter edge-to-edge repair (TEER). During the operation, mitral leaflets capture and clamping have the highest requirements for ultrasound image quality and should be performed under the guidance of high-quality commissural view. However, standard commissural view cannot be obtained in some patients due to cardiac enlargement, transposition or limited esophageal acoustic window. In this condition, the optimal view can be obtained by Multivue technology. This paper reports a male patient aged 67 years with successful mitral valve TEER under the real-time guidance of Multivue technology, and summarizes the key points of this technology.

Pancreatic cancer is highly aggressive and often diagnosed at an advanced stage,leaving most patients ineligible for radical resection.This study retrospectively analyzed four patients with locally advanced or advanced pancreatic cancer to evaluate the clinical efficacy and safety of high-intensity focused ultrasound(HIFU)ablation combined with chemotherapy as a neoadjuvant and conversion therapy.All cases were reviewed and individualized treatment plans were formulated through a multidisciplinary team evaluation.All patients received HIFU plus gemcitabine and nab-paclitaxel chemotherapy,with assessments of tumor volume,vascular involvement,surgical conversion,symptom relief,and adverse events.Three patients achieved marked tumor shrinkage and reduction of vascular invasion,enabling successful R0 resection without recurrence during follow-up.The remaining patient achieved disease stability,significant pain relief,and maintained good quality of life under repeated HIFU therapy.All treatments were well tolerated,and no severe adverse reactions occurred.The combination of HIFU and chemotherapy demonstrated synergistic local and systemic effects,effectively achieving tumor downstaging,improving resectability,and alleviating symptoms.As a safe,noninvasive,and repeatable therapeutic approach,this strategy offers a promising option for patients with advanced pancreatic cancer.Further large-scale prospective studies are warranted to validate its long-term efficacy and elucidate underlying mechanisms.

Objective:To explore the clinical efficacy of membrane induction technique in the treatment of chronic bone infection after internal fixation of intertrochanteric fracture in adults.Methods:The data were retrospectively analyzed of the 15 adult patients who had been treated for chronic bone infection after internal fixation of intertrochanteric fracture at Department of Traumatic Orthopaedics Ⅱ, Gansu Provincial People's Hospital from May 2019 to May 2022. There were 13 males and 2 females with an age of (55.8±2.6) years. By the Cierny-Mader anatomical classification, 6 cases were type Ⅲ and 9 cases type Ⅳ; by the physiological classification, 10 cases were type A and 5 cases type B. Course of bone infection was (22.1±8.1) months. All patients were treated by membrane induction technique in 2 stages. In the first stage, antibiotic bone cement was implanted after debridement into the defects [(4.50±0.54) cm in length] followed by temporary fixation of broken ends of the fracture. After 6 to 8 weeks when infection was controlled, the second stage repair and reconstruction of bone defects was carried out. Infection control, complications, bone healing time, and Samantha X-ray bone healing score, visual analogue scale (VAS) at standing, Sanders hip score and Paley score for bone defect healing at the last follow-up were recorded.Results:The 15 patients were followed up for (27.5±2.5) months. Infection recurred after the first-stage operation in 2 cases, but was effectively controlled after debridement and follow-up observed no recurrence. Follow-up showed no such complications as donor site infection, deformation or collapse of the bone reconstructed, limb shortening, re-fracture or internal fixation failure in the 15 patients who achieved bony union after (10.3±2.1) months. At the last follow-up, the Samantha X-ray score was 6.0(5.0, 6.0) points and VAS at standing 1.0(1.0, 3.0) point for the 15 patients; by Sanders hip score, the hip function was rated as excellent in 10 cases and as fine in 5 cases; by Paley score for bone defect healing, the efficacy was rated as excellent in 7 cases, as good in 2 cases and as fair in 6 cases.Conclusion:In the treatment of chronic bone infection after internal fixation of intertrochanteric fracture in adults, membrane induction technique can effectively control infection and improve the hip function of the affected limb, leading to good clinical efficacy.

Objective:To explore the clinical efficacy of membrane induction technique in the treatment of chronic bone infection after internal fixation of intertrochanteric fracture in adults.Methods:The data were retrospectively analyzed of the 15 adult patients who had been treated for chronic bone infection after internal fixation of intertrochanteric fracture at Department of Traumatic Orthopaedics Ⅱ, Gansu Provincial People's Hospital from May 2019 to May 2022. There were 13 males and 2 females with an age of (55.8±2.6) years. By the Cierny-Mader anatomical classification, 6 cases were type Ⅲ and 9 cases type Ⅳ; by the physiological classification, 10 cases were type A and 5 cases type B. Course of bone infection was (22.1±8.1) months. All patients were treated by membrane induction technique in 2 stages. In the first stage, antibiotic bone cement was implanted after debridement into the defects [(4.50±0.54) cm in length] followed by temporary fixation of broken ends of the fracture. After 6 to 8 weeks when infection was controlled, the second stage repair and reconstruction of bone defects was carried out. Infection control, complications, bone healing time, and Samantha X-ray bone healing score, visual analogue scale (VAS) at standing, Sanders hip score and Paley score for bone defect healing at the last follow-up were recorded.Results:The 15 patients were followed up for (27.5±2.5) months. Infection recurred after the first-stage operation in 2 cases, but was effectively controlled after debridement and follow-up observed no recurrence. Follow-up showed no such complications as donor site infection, deformation or collapse of the bone reconstructed, limb shortening, re-fracture or internal fixation failure in the 15 patients who achieved bony union after (10.3±2.1) months. At the last follow-up, the Samantha X-ray score was 6.0(5.0, 6.0) points and VAS at standing 1.0(1.0, 3.0) point for the 15 patients; by Sanders hip score, the hip function was rated as excellent in 10 cases and as fine in 5 cases; by Paley score for bone defect healing, the efficacy was rated as excellent in 7 cases, as good in 2 cases and as fair in 6 cases.Conclusion:In the treatment of chronic bone infection after internal fixation of intertrochanteric fracture in adults, membrane induction technique can effectively control infection and improve the hip function of the affected limb, leading to good clinical efficacy.

Pancreatic cancer is highly aggressive and often diagnosed at an advanced stage,leaving most patients ineligible for radical resection.This study retrospectively analyzed four patients with locally advanced or advanced pancreatic cancer to evaluate the clinical efficacy and safety of high-intensity focused ultrasound(HIFU)ablation combined with chemotherapy as a neoadjuvant and conversion therapy.All cases were reviewed and individualized treatment plans were formulated through a multidisciplinary team evaluation.All patients received HIFU plus gemcitabine and nab-paclitaxel chemotherapy,with assessments of tumor volume,vascular involvement,surgical conversion,symptom relief,and adverse events.Three patients achieved marked tumor shrinkage and reduction of vascular invasion,enabling successful R0 resection without recurrence during follow-up.The remaining patient achieved disease stability,significant pain relief,and maintained good quality of life under repeated HIFU therapy.All treatments were well tolerated,and no severe adverse reactions occurred.The combination of HIFU and chemotherapy demonstrated synergistic local and systemic effects,effectively achieving tumor downstaging,improving resectability,and alleviating symptoms.As a safe,noninvasive,and repeatable therapeutic approach,this strategy offers a promising option for patients with advanced pancreatic cancer.Further large-scale prospective studies are warranted to validate its long-term efficacy and elucidate underlying mechanisms.

Objectives:To evaluate the valvular and cardiac function,cardiac reverse remodeling at 6-month after transcatheter edge-to-edge repair(TEER)for patients with functional and degenerative mitral valve regurgitation,and summarize the experience of echocardiography application. Methods:The clinical data of 93 patients with moderate to severe mitral regurgitation(MR)treated with TEER and completed 6-month follow-up in Yunnan Fuwai Cardiovascular Hospital from July 2022 to February 2023 were retrospectively analyzed.Patients were divided into functional mitral regurgitation(FMR)and degenerative mitral regurgitation(DMR)groups according to MR etiology.The valve characteristic parameters,as well as valvular function,chamber volume and cardiac functional parameters before and at 6 months after operation were compared.The key points of echocardiography application were summarized. Results:Among all patients,71 were FMR and 22 were DMR.There were differences in valve structure between the two groups.Mitral TEER were successfully accomplished and all patients completed 6-month follow-up.The key points of echocardiography application included:valve structure analysis,atrial septal puncture location,device delivery process monitoring and image optimization during clamping process.The mitral regurgitation grade and NYHA grade were significantly improved in all patients at 6 months after TEER(P<0.05),and the mean mitral valve pressure gradient was higher than that before operation(P<0.05).Left ventricular end-diastolic volume(LVEDV),left ventricular end-systolic volume(LVESV)and left atrial volume index in FMR group were significantly decreased(P<0.05),while left ventricular and left atrial volume in DMR group remained unchanged(P>0.05).There were no significant changes in left ventricular ejection fraction and left ventricular global strain in both groups during the observation period(P>0.05).The changes of LVEDV and LVESV before and after operation were more significant in FMR group than those in DMR group(P<0.05). Conclusions:Mitral TEER can reduce the degree of regurgitation and improve cardiac function in the early postoperative period for moderate and severe MR patients with different etiologies.There are differences in preoperative valve structure and postoperative cardiac reverse remodeling between FMR and DMR patients.Echocardiography is an important imaging technique for the evaluation and monitoring process before,during and post mitral TEER.

[Objective]To evaluate the knowledge,attitude and practice(KAP)of medication use and safety among the patients with acquired immune deficiency syndrome(AIDS)in a 3A-grade hospital of Guangzhou city,and to provide scientific basis for AIDS prevention and treatment.[Methods]A questionnaire survey was conducted among AIDS patients aged 18 years and above in our hospital to investigate their KAP regarding medication use and safety.[Results]A total of 549 questionnaires were collected,of which 503 were valid,with an effective recovery rate of 91.6%.The average scores of KAP were(14.58±8.49),(25.21±6.92)and(47.58±3.33),respectively,with the scoring rates of 36.46%,63.02%,and 95.16%,respectively.There were statistically significant differences(P＜0.05)in knowledge scores among people with different ages,education levels and occupations.Multiple linear regression showed that education level and medical insurance status had most significant impact on knowledge scores(P<0.05).Significant differences were found in attitude scores among people with different education levels(P<0.05),as well as in practice scores among people with different occupations(P<0.05).Multiple linear regression revealed that age,occupation,knowledge score and attitude score had a significant impact on practice scores(P<0.05).Patients expected to receive pharmaceutical care services from the pharmacists via face-to-face communication,network platform and telephone consultation on medication knowledge such as adverse drug reactions and response measures,drug-drug interactions,missed medication and response measures,medication adherence measures,etc.[Conclusions]AIDS patients in this hospital have a good awareness of medication safety,but their knowledge of medication use needs improvement.Some bad habits may affect their compliance,resulting in safety hazards.Therefore,there is an urgent demand for pharmaceutical care services related to rational drug use.

Objective:To utilize routinely available clinical parameters to uncover the clinical features of different clusters in patients with chronic rhinosinusitis with nasal polyp (CRSwNP) through unsupervised clustering analysis.Methods:The clinical data from 155 CRSwNP patients undergoing nasal endoscopic surgery at Renmin Hospital of Wuhan University from 2021 to 2023 were prospectively collected, including 112 males and 43 females, aged from 7 to 87 years. Unsupervised clustering analysis was conducted using various clinical parameters, including age, gender, smoking and drinking history, local eosinophil (EOS) and neutrophil (NEU) counts, comorbid allergic rhinitis (AR), comorbid asthma, recurrence status, serum-specific IgE, total IgE, cytokine levels, peripheral blood EOS count and percentage, Lund-Mackay CT score, the ratio of CT scores for the maxillary sinus and ethmoid sinus (E/M ratio), visual analogue scale (VAS) score, Lund-Kennedy endoscopic score, and other common clinical indicators to elucidate the clinical characteristics of each cluster. Statistical analysis was conducted using GraphPad Prism 9.5 software.Results:Hierarchical clustering analysis identified four main clusters (Cluster A1-A4), showcasing distinct characteristics such as mild nasal polyps with higher peripheral blood cytokines levels, nasal polyps accompanied by allergies and asthma, a subtype of nasal polyps with high recurrence rates dominated by neutrophils, and nasal polyps with high eosinophil levels. Further subset clustering revealed two clusters of mild polyps (Cluster B1-B2) featuring high cytokine expression and comorbid AR; and two clusters of severe polyps (Cluster B3-B4) presented with severe symptoms, high Lund-Mackay CT score, and high Lund-Kennedy endoscopic score. Variations between Cluster B3 and B4 included symptom complexity, the degree of eosinophil infiltration, and the probability of comorbid asthma. Further clustering analysis for eosinophilic nasal polyps revealed a cluster characterized by highly neutrophilic infiltration and recurrent nasal polyps. The comprehensive analysis of multi-index correlations demonstrated valuable insights into the relationships between common clinical parameters of nasal polyps, providing valuable information for a deeper understanding of the pathogenesis of CRSwNP.Conclusion:The clustering analysis in this study categorizes CRSwNP patients into different clusters based on clinical features and disease outcomes, providing a new perspective for more precise clinical treatment strategies.

Background::Pulmonary arterial hypertension (PAH) is characterized by excessive proliferation of small pulmonary arterial vascular smooth muscle cells (PASMCs), endothelial dysfunction, and extracellular matrix remodeling. G protein-coupled receptor kinase 2 (GRK2) plays an important role in the maintenance of vascular tone and blood flow. However, the role of GRK2 in the pathogenesis of PAH is unknown.Methods::GRK2 levels were detected in lung tissues from healthy people and PAH patients. C57BL/6 mice, vascular smooth muscle cell-specific Grk2-knockout mice ( Grk2?SM22), and littermate controls ( Grk2flox/flox) were grouped into control and hypoxia mice ( n = 8). Pulmonary hypertension (PH) was induced by exposure to chronic hypoxia (10%) combined with injection of the SU5416 (cHx/SU). The expression levels of GRK2 and Yes-associated protein (YAP) in pulmonary arteries and PASMCs were detected by Western blotting and immunofluorescence staining. The mRNA expression levels of Grk2 and Yes-associated protein ( YAP) in PASMCs were quantified with real-time polymerase chain reaction (RT-PCR). Wound-healing assay, 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide (MTT) assay, and 5-Ethynyl-2′-deoxyuridine (EdU) staining were performed to evaluate the proliferation and migration of PASMCs. Meanwhile, the interaction among proteins was detected by immunoprecipitation assays. Results::The expression levels of GRK2 were upregulated in the pulmonary arteries of patients with PAH and the lungs of PH mice. Moreover, cHx/SU-induced PH was attenuated in Grk2?SM22 mice compared with littermate controls. The amelioration of PH in Grk2?SM22 mice was accompanied by reduced pulmonary vascular remodeling. In vitro study further confirmed that GRK2 knock-down significantly altered hypoxia-induced PASMCs proliferation and migration, whereas this effect was severely intensified by overexpression of GRK2. We also identified that GRK2 promoted YAP expression and nuclear translocation in PASMCs, resulting in excessive PASMCs proliferation and migration. Furthermore, GRK2 is stabilized by inhibiting phosphorylating GRK2 on Tyr86 and subsequently activating ubiquitylation under hypoxic conditions. Conclusion::Our findings suggest that GRK2 plays a critical role in the pathogenesis of PAH, via regulating YAP expression and nuclear translocation. Therefore, GRK2 serves as a novel therapeutic target for PAH treatment.