This study established the mouse podocyte clone-5 (MPC5) with Smad3 knockout and studied the effect of transforming growth factor-beta 1 (TGF-β1) on the dedifferentiation of the MPC5 cells with Smad3 knockout, aiming to provide a cell tool for studying the role of Smad3 in mouse podocytes. The single-guide RNA (sgRNA) sequence targeting Smad3 was designed according to the principles of CRISPR/Cas9 design. The pX458-Smad3 vector was constructed and introduced into competent cells, and then the vector was extracted and used to transfect MPC5 cells. The successfully transfected cells were sorted by a flow cytometer. After single-cell clone expansion, PCR amplification of sequences adjacent to the edition site of Smad3 and sequencing were performed to identify potential cells with gene knockout. Western blotting was employed to verify the knockout efficiency of Smad3. Finally, the effect of Smad3 knockout on TGF-β1-induced dedifferentiation of MPC5 cells was analyzed by reverse transcription-polymerase chain reacting (RT-PCR), Western blotting, and the immunofluorescence method. The sgRNA was designed to target the fifth exon of Smad3. EGFP expression was observed 24 h after transfection of the pX458-Smad3 plasmid into MPC5 cells, with the transfection efficiency of 0.1% as determined by flow cytometry. From the transfected cells, 21 cell clones were obtained through flow cytometric sorting and single-cell clone expansion. PCR amplification and sequencing of the region around the sgRNA target site in Smad3 identified two cell clones with biallelic frameshift mutations. Western blotting results confirmed the absence of Smad3 expression in these clones, indicating successful establishment of the MPC5 cell line with Smad3 knockout. In normal MPC5 cells, TGF-β1 stimulation promoted the expression of fibrosis-related genes fibronectin and Col1a1 (collagen I) and inhibited the expression of the podocyte marker proteins synaptopodin and podocin, which suggested epithelial-mesenchymal transition and podocyte injury. However, in the two MPC5 cell lines with Smad3 knockout, TGF-β1-induced expression of epithelial-mesenchymal transition markers was significantly suppressed. The MPC5 cell lines with Smad3 knockout that were constructed by CRISPR/Cas9 provide a valuable cell model for functional studies of Smad3 protein and highlight the critical role of Smad3 in cell dedifferentiation.
Animals ; Smad3 Protein/genetics* ; CRISPR-Cas Systems/genetics* ; Mice ; Podocytes/metabolism* ; Transforming Growth Factor beta1/pharmacology* ; Cell Line ; Gene Knockout Techniques ; RNA, Guide, CRISPR-Cas Systems/genetics*

Objective To analyze the international research hotspots and development trends of social work intervention in chronic disease management,providing reference for domestic research on social work intervention in chronic disease manage-ment.Methods The research literature was retrieve on the core collection of Web of Science,articles published from the estab-lishment of the database from 2000 to 2024.CiteSpace were used to present the keyword co-occurrence,keyword clustering,au-thor collaboration network,and institutional collaboration network of social work intervention in chronic disease management.Results The post-documentation situation showed a fluctuating upward trend as a whole.The keywords that appeared frequently were"social support""management""quality of life""chronic disease"and"Care".There was multi-agency and multi-author collaboration.Conclusion Internationally,social work intervention in chronic disease management focuses on social support and quality of life.The research cooperation system in related fields is diversified,and social workers play an active role in the inter-disciplinary teams for chronic disease management.

Objective To analyze the international research hotspots and development trends of social work intervention in chronic disease management,providing reference for domestic research on social work intervention in chronic disease manage-ment.Methods The research literature was retrieve on the core collection of Web of Science,articles published from the estab-lishment of the database from 2000 to 2024.CiteSpace were used to present the keyword co-occurrence,keyword clustering,au-thor collaboration network,and institutional collaboration network of social work intervention in chronic disease management.Results The post-documentation situation showed a fluctuating upward trend as a whole.The keywords that appeared frequently were"social support""management""quality of life""chronic disease"and"Care".There was multi-agency and multi-author collaboration.Conclusion Internationally,social work intervention in chronic disease management focuses on social support and quality of life.The research cooperation system in related fields is diversified,and social workers play an active role in the inter-disciplinary teams for chronic disease management.

Objective To explore the relationship between preoperative coronary angiography and postoperative acute kidney injury (AKI) in cardiac surgery. Methods The clinical data of patients who underwent coronary angiography within 30 days before cardiac surgery in the First Affiliated Hospital of Xi’an Jiaotong University from January 2015 to April 2019 were retrospectively analyzed. Univariate analysis and multivariate logistic regression analyses were used to explore the relationship between the interval from preoperative coronary angiography to cardiac surgery and postoperative AKI. Results Finally 1 112 patients were collected, including 700 males and 412 females, with a median age of 61 (55, 66) years. The incidence of postoperative AKI was 40.8% (454/1 112), of which grade 2-3 AKI accounted for 11.9%. Multivariate analysis showed that age (OR=1.049, 95%CI 1.022-1.077, P<0.001), body mass index (OR=1.065, 95%CI 1.010-1.123, P=0.020) and time interval between preoperative coronary angiography and cardiac surgery within 24 hours (OR=1.625, 95%CI 1.116-2.364, P=0.011) were independent predictors of postoperative AKI. Patients who underwent coronary angiography within 24 hours before surgery had a 10.6% higher incidence of postoperative AKI compared to those who underwent angiography ≥24 hours before surgery (P=0.004). Patients who underwent valve surgery with or without coronary artery bypass grafting (CABG) had a higher risk of AKI than those who only underwent CABG. The in-hospital stay of patients who developed AKI was 2 days longer than those without AKI. However, undergoing coronary angiography within 24 hours before cardiac surgery did not prolong the length of ICU stay or hospital stay, nor did it increase the risk of death or renal failure after the operation. Conclusion Undergoing coronary angiography within 24 hours before cardiac surgery increases the risk of postoperative AKI.

Objective:To identify risk variables for prolonged postoperative length of stay (PPOLOS) in hip fracture patients by machine learning algorithms and construct Nomogram models.Methods:A retrospective case-control study was conducted to select 248 patients with hip fracture diagnosed and treated in Yingtan 184th Hospital from June 2019 to June 2023 by convenient sampling method. Two machine learning algorithms were used (least absolute shrinkage and selection operator, LASSO and support vector machine-Recursive Feature Elimination, SVM-RFE) to screen PPOLOS risk variables. Construct a Nomogram model to predict the risk of PPOLOS in patients with hip fracture based on intersection risk variables. The model was validated using internal data sets.Results:Among the 248 patients with hip fracture, there were 79 males and 169 females, aged (64.49 ± 8.02). The mean postoperative length of hospital stay of 248 patients was (7.98 ± 5.68) d, and the median was 7 d. LASSO algorithm identifies 7 risk variables, the SVM-RFE algorithm identified 8 risk variables. Intersectional risk variables were age, body mass index (BMI), Charlson comorbidity index (CCI), type of surgery, and central granulocytocyte ratio (NLR). Multivariate Logistic regression analysis(intersection risk variables) showed that age [ OR=1.649(1.235-2.202)], BMI [1.603(1.204-2.134)], CCI [ OR=1.670(1.236-2.258)], type of surgery [ OR=1.620(1.209-2.170), 1.699(1.243-2.321)], and NLR [ OR=3.258(2.299-4.617)] were independently associated with the risk of PPOLOS (all P<0.05). The results showed that the conformity index of the Nomogram model was 0.865 (95% CI=0.768-0.945). The area under the curve was 0.852 (95% CI=0.748-0.962). When the risk threshold was >0.08, it could provide significant clinical net benefit. The clinical impact curve showed effective identification of PPOLOS patients in high-risk groups. Conclusions:This Nomogram model can guide medical staff to make clinical diagnosis and treatment decisions as soon as possible to avoid risks, allocate medical resources rationally, and improve nursing quality.

Objective To investigate the therapeutic effect and underlying mechanism of Qishishenshu Capsule on renal fibrosis in mice with early diabetic nephropathy(DN).Methods A DN mouse model was established by multiple injections of streptozotocin.The mice were randomly divided into a normal group(NC),model group(DN),and Qishi group(QS)(0.9 g/(kg·d)),with eight mice in each group.Mice were gavaged continuously for 4 weeks,and fasting blood glucose(FBG)was measured weekly.Four weeks later,urinary albumin/creatinine(UACR),serum creatinine,and blood urea nitrogen were measured.Hematoxylin-eosin,periodicacid-Schiff,and Sirius red staining were used to analyze renal pathological changes.Real-time fluorescence quantitative reverse-transcription polymerase chain reaction was used to detect the mRNA levels of fibronectin(FN),collagen type Ⅰ alpha 1(Col1a1),and α-smooth muscle actin(α-SMA).Immunohistochemistry and Western blot were performed to detect FN,collagen type Ⅰ(Collagen Ⅰ),collagen typeⅢ(Collagen Ⅲ),α-SMA,Podocin,Nephrin,and transforming growth factor-β1/SMAD family member2/3(TGF-β1/Smad2/3)pathway-related proteins.Results Compared with mice in the NC group,those in the DN group showed significantly higher levels of FBG and UACR(P＜0.001),and mesangial hyperplasia,basement membrane thickening,and collagen deposition in the renal tissue.The mRNA levels of FN,Col1a1,and α-SMA were increased(P＜0.05).Protein levels of Podocin and Nephrin were decreased(P＜0.05).The levels of FN,Collagen I,Collagen Ⅲ,α-SMA,and TGF-β1/Smad2/3 pathway proteins were increased(P＜0.05).Compared with the DN group,the QS group's level of UACR was decreased(P＜0.05),their renal pathological injury was alleviated,and mRNA levels of FN,Collagen Ⅰ,andα-SMA were attenuated(P＜0.05);whereas their protein levels of Podocin and Nephrin were elevated(P＜0.05).The levels of FN,Collagen Ⅰ,Collagen Ⅲ,α-SMA,and TGF-β1/Smad2/3 pathway proteins were also decreased(P＜0.05).Conclusions Qishishenshu Capsule improved renal fibrosis in DN mice,probably through the inhibition of the TGF-β1/Smad2/3 signaling pathway.

Objective:This study proposed a novel technique for aortic root reconstruction and evaluated its safety and effectiveness.Methods:Acute type A aortic dissection (ATAAD) patients who underwent Sun' s procedure from January 2020 to June 2022 in the First Affiliated Hospital of Xi'an Jiaotong University were retrospectively enrolled. These patients were divided into three groups according to their surgical procedures of aortic root: aortic root reinforcement combined with vascular grafts built-in and intermittent anastomosis technique (XJ-procedure) group, continuous suture and pledgeted interrupted reinforcement group and Bentall group. The perioperative data and postoperative follow-up results of the three groups were compared to analyze the efficacy of XJ-procedure in the root treatment of ATAAD.Results:A total of 509 patients were enrolled in this study, including 288 patients in the XJ-procedure group, 132 patients in the continuous suture and pledgeted interrupted reinforcement group and 89 patients in the Bentall group. The 30-day mortality of all patients was 7.5% (38/509), and were 7.3%, 8.3% and 6.7% in the XJ-procedure group, continuous suture and pledgeted interrupted reinforcement group and Bentall group, respectively. The operation time of the XJ-procedure group [355 (320, 380) min] was significantly lower than the other two groups. The cardiopulmonary bypass time [154 (140, 169) min] and the aortic cross-clamping time [80 (72, 89) min] of XJ-procedure group were lower than the Bentall group [166 (147, 184) min and 86 (77, 96) min]. The aortic root bleeding that required secondary cardiopulmonary bypass did not occur in the XJ-procedure group. There was no significant difference in early postoperative complications among the three groups. The incidence of residual aortic root dissection before discharge, in 3 and 6 months after operation was lower in the XJ-procedure group compared to the continuous suture and pledgeted interrupted reinforcement group ( P<0.001). Conclusion:The XJ-procedure technique is a safe, simple and mastered method for the aortic root treatment of ATAAD, which shows an accurate early clinical effect.

Objective:This study proposed a novel technique for aortic root reconstruction and evaluated its safety and effectiveness.Methods:Acute type A aortic dissection (ATAAD) patients who underwent Sun' s procedure from January 2020 to June 2022 in the First Affiliated Hospital of Xi'an Jiaotong University were retrospectively enrolled. These patients were divided into three groups according to their surgical procedures of aortic root: aortic root reinforcement combined with vascular grafts built-in and intermittent anastomosis technique (XJ-procedure) group, continuous suture and pledgeted interrupted reinforcement group and Bentall group. The perioperative data and postoperative follow-up results of the three groups were compared to analyze the efficacy of XJ-procedure in the root treatment of ATAAD.Results:A total of 509 patients were enrolled in this study, including 288 patients in the XJ-procedure group, 132 patients in the continuous suture and pledgeted interrupted reinforcement group and 89 patients in the Bentall group. The 30-day mortality of all patients was 7.5% (38/509), and were 7.3%, 8.3% and 6.7% in the XJ-procedure group, continuous suture and pledgeted interrupted reinforcement group and Bentall group, respectively. The operation time of the XJ-procedure group [355 (320, 380) min] was significantly lower than the other two groups. The cardiopulmonary bypass time [154 (140, 169) min] and the aortic cross-clamping time [80 (72, 89) min] of XJ-procedure group were lower than the Bentall group [166 (147, 184) min and 86 (77, 96) min]. The aortic root bleeding that required secondary cardiopulmonary bypass did not occur in the XJ-procedure group. There was no significant difference in early postoperative complications among the three groups. The incidence of residual aortic root dissection before discharge, in 3 and 6 months after operation was lower in the XJ-procedure group compared to the continuous suture and pledgeted interrupted reinforcement group ( P<0.001). Conclusion:The XJ-procedure technique is a safe, simple and mastered method for the aortic root treatment of ATAAD, which shows an accurate early clinical effect.

OBJECTIVE To establish a method for simultaneous determination of atorvastatin （ATV） and its active metabolites 2-hydroxy atorvastatin acid （2-HAT）， 4-hydroxy atorvastatin acid （4-HAT） and toxic metabolite atorvastatin lactone （ALT） in rat plasma and apply it for pharmacokinetic study. METHODS LC-MS/MS method was adopted for analysis. The one-step precipitation method was used for processing plasma samples （plasma samples were pretreated by acidification to adjust pH value so as to prevent inversion of configuration）， gradient elution was used to analyze the samples， and the analysis time was 5 min. Electrospray positive ionization was adopted， and positive ion scanning was performed in multi-reaction monitoring. The m/z of quantified ion pairs of ATV and its metabolites such as 2-HAT， 4-HAT and ATL， and internal standard pitavastatin were 559.3→ 440.2， 575.2→440.3， 575.0→440.2， 540.9→448.2 and 422.2→290.0， respectively. After conducting a comprehensive methodological investigation of the analytical method， the concentrations of ATV and its metabolites 2-HAT， 4-HAT，and ATL were determined， and the pharmacokinetic parameters of ATV and its metabolites were calculated using the non- compartment model of WinNonlin 6.1. RESULTS The results of methodological validation showed that endogenous substances in blank plasma did not interfere with the determination of the components to be tested， and the standard curve had a good linear relationship； the lower limits of quantification for ATV， 2-HAT， 4-HAT and ATL were 0.5， 0.5， 0.25 and 0.063 nmol/L， respectively. The precision， accuracy， recovery， matrix effect and stability investigation were all in line with the requirements of biological analysis. Pharmacokinetic analysis showed that after intragastric administration in rats， ATV calcium metabolized rapidly， and was mainly exposed to blood circulation in the form of ATV and 2-HAT， with the lowest concentration of lactone-type metabolites. CONCLUSIONS The established method is precise， rapid and accurate for plasma concentration analysis of ATV and its active/toxic metabolites. The application of the method could help to fully elucidate the pharmacokinetic characteristics of atorvastatin calcium in rats.

Objective To extract and summarize the best evidence of manual rehabilitation therapy in patients with breast cancer related lymphedema.It aims to provide evidence-based basis for clinical nursing staff interven-tion.Methods According to the"6S evidence resource pyramid model",we systematically retrieved from UpTo-Date,BMJ best clinical practice,JBI evidence-based medicine center database of Joanna Briggs Institute in Australi-a,Embase,Cochrane Library,Pubmed,GIN,New Zealand Guidelines collaboration group,SIGN,American Cancer So-ciety,American Association for Physical Therapy,American Society for Lymphatic,American Lymphedema Website,Ontario Association of Registered Nurses of Canada,Yimaitong,CNKI,VIP,CBM,WanFang,CHINAL,Web of Science and OVID databases from establishment of database to January 2023.It mainly includes guidelines,consensuses,evi-dence summaries,clinical decision-making,systematic evaluation and primitive research.There were 2 researchers who independently evaluated the quality of the literature and extracted evidence.Results A total of 19 pieces of the literature were involved,including 4 guidelines,4 consensuses,3 system reviews,1 recommended practice,2 clinical decision-making,1 summary of evidence,4 primitive research.Finally,26 pieces of best evidence are summarized from the 5 aspects of indications,contraindications,treatment points,treatment effects and precautions of manual re-habilitation.Conclusion This study summarized the evidence of manual rehabilitation therapy in breast cancer re-lated lymphedema.It can provide practical basis for clinical nursing staff.In clinical practice,it is important to strictly evaluate whether patients have contraindications to treatment,to follow the recommended treatment plan and monitor complications during treatment.