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 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: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 evaluate the effect of preemptive analgesia with ibuprofen on postoperative pain following single posterior tooth implantation, aiming to provide a clinical reference for its application.Methods:A multicenter, randomized, double-blind, placebo-controlled parallel-group trial was conducted. A total of 82 participants were included in the trial, meeting the eligibility criteria from April 2022 to April 2024 at the Capital Medical University School of Stomatology (40 cases), Beijing TianTan Hospital, Capital Medical University (22 cases), Beijing Chao-Yang Hospital, Capital Medical University (20 cases). Participants were randomly assigned in a 1∶1 ratio to either the ibuprofen group or the control group, with each group comprising 41 individuals. Participants in the ibuprofen group received 300 mg of sustained-release ibuprofen capsules orally 15 min before surgery, while the control group received a placebo. Both groups received the same postoperative analgesic regimen for 3 days. Pain scores were assessed using the numerical rating scale at 30 min, 4 h, 6 h, 8 h, 24 h, 48 h, and 72 h postoperatively, and the additional use of analgesic medication was recorded from days 4 to 6 postoperatively.Results:A total of 82 participants were initially enrolled in the study, with 7 dropouts (4 from the control group and 3 from the ibuprofen group), resulting in 75 participants (37 in the control group and 38 in the ibuprofen group) completing the trial. There were no reports of adverse events such as nausea or vomiting among the participants. The ibuprofen group exhibited significantly lower pain scores at 4 h, 6 h and 8 h [1.0 (0.0, 2.0), 1.0 (0.0, 2.0), 1.5 (0.0, 3.0) ] postoperatively compared to the control group 4 h, 6 h and 8 h [2.0 (1.0, 3.0), 3.0 (1.5, 4.0), 2.0 (1.0, 4.0)] ( Z=-1.99, P=0.047; Z=-3.01, P=0.003; Z=-2.10, P=0.036). The proportions of patients requiring additional analgesic medication between days 4 and 6 post-surgery were 18.4% (7/38) in the ibuprofen group and 27.0% (10/37) in the control group, with no significant difference (χ 2=0.79, P=0.373). The median additional medication usage postoperatively was [0.0 (0.0, 0.0) pills] in the ibuprofen group and [0.0 (0.0, 1.0) pills] in the control group, with no significant difference ( Z=-0.78, P=0.439). Conclusions:Preemptive analgesia with ibuprofen effectively reduces postoperative pain following tooth implantation, representing a safe and effective perioperative pain management strategy.

Objective:To systematically review the prevalence, influencing factors of financial toxicity and coping strategies in cancer patients.Methods:The studies on financial toxicity among cancer patients were searched in PubMed, Embase, The Cochrane Library, Web of Science, CINAHL, WangFang Data, CNKI, Sinomed, VIP from inception to June 2022. Stata14.0 software was used to analyze the prevalence of financial toxicity and coping strategies with a single group meta-analysis. Descriptive analysis was used to analyze the influencing factors.Results:A total of 25 papers including 2 939 cancer patients were included. Meta-analysis results showed that the prevalence of financial toxicity in cancer patients was 47% (95% CI 39%-55%). The influencing factors of financial toxicity included sociodemographic factors, disease related factors, medical insurance related factors and psychological factors. Conclusions:The high incidence of financial toxicity in cancer patients is associated with a variety of factors. Health care professionals should pay attention to early screening of cancer patients, dynamically collect relevant data such as patient expenditure items, continuously assess patients' economic status, and make relevant preventive efforts.

Background::Erythropoietin is a glycoprotein that mainly regulates erythropoiesis. In patients with chronic renal failure with anemia, darbepoetin alfa can stimulate erythropoiesis, correct anemia, and maintain hemoglobin levels. This study was designed to demonstrate the efficacy and safety of darbepoetin alfa injections as being not inferior to epoetin alfa injections (Recombinant Human Erythropoietin injection, rHuEPO) when maintaining hemoglobin (Hb) levels within the target range (10.0-12.0 g/dL) for the treatment of renal anemia.Methods::Ninety-five patients were enrolled in this study from April 15, 2013 to April 10, 2014 at 25 sites. In this study, patients ( n = 95) aged 18-70 years were randomized into a once per week intravenous darbepoetin alfa group ( n = 56) and a twice or three times per week intravenous epoetin alfa group ( n = 39) for 28 weeks, who had anemia with hemoglobin levels between 6 g/dL and 10 g/dL due to chronic kidney disease (CKD) and were undergoing hemodialysis or hemofiltration with ESA-naive (erythropoiesis stimulating agent-naive). The primary efficacy profile was the mean Hb level (the non-inferiority margin was -1.0 g/dL, week 21-28); the secondary efficacy profiles were the Hb increase rate (week 0-4), the target Hb achievement cumulative rate and time, the change trends of the Hb levels, and the target Hb maintenance ratio. Adverse events (AEs) were observed and compared, and the efficacy and safety were analyzed between the two treatment groups. Additionally, the frequencies of dose adjustments between the darbepoetin alfa and epoetin alfa groups were compared during the treatment period. SAS? software version 9.2 was used to perform all statistical analyses. Descriptive statistics were used for all efficacy, safety, and demographic variable analyses, including for the primary efficacy indicators. Results::The mean Hb level was 11.3 g/dL in the darbepoetin alfa group and 10.7 g/dL in the epoetin alfa group, respectively; the difference of the lower limits of the 95% confidence intervals (CI) between the two groups was 0.1 g/dL (>-1.0 g/dL), and non-inferiority was proven; the Hb levels started to increase in the first four weeks at a similar increase rate; no obvious differences were observed between the groups in the target Hb achievement cumulative rates, and the Hb levels as well as the target Hb level maintenance rate changed over time. The incidence of AEs was 62.5% in the darbepoetin alfa group and 76.9% in the epoetin alfa group. All the adverse events observed in the study were those commonly associated with hemodialysis.Conclusion::Darbepoetin alfa intravenously once per week can effectively increase Hb levels and maintain the target Hb levels well, which makes it not inferior to epoetin alfa intravenously twice or three times per week. Darbepoetin alfa shows an efficacy and safety comparable to epoetin alfa for the treatment of renal anemia.

Background::Erythropoietin is a glycoprotein that mainly regulates erythropoiesis. In patients with chronic renal failure with anemia, darbepoetin alfa can stimulate erythropoiesis, correct anemia, and maintain hemoglobin levels. This study was designed to demonstrate the efficacy and safety of darbepoetin alfa injections as being not inferior to epoetin alfa injections (Recombinant Human Erythropoietin injection, rHuEPO) when maintaining hemoglobin (Hb) levels within the target range (10.0-12.0 g/dL) for the treatment of renal anemia.Methods::Ninety-five patients were enrolled in this study from April 15, 2013 to April 10, 2014 at 25 sites. In this study, patients ( n = 95) aged 18-70 years were randomized into a once per week intravenous darbepoetin alfa group ( n = 56) and a twice or three times per week intravenous epoetin alfa group ( n = 39) for 28 weeks, who had anemia with hemoglobin levels between 6 g/dL and 10 g/dL due to chronic kidney disease (CKD) and were undergoing hemodialysis or hemofiltration with ESA-naive (erythropoiesis stimulating agent-naive). The primary efficacy profile was the mean Hb level (the non-inferiority margin was -1.0 g/dL, week 21-28); the secondary efficacy profiles were the Hb increase rate (week 0-4), the target Hb achievement cumulative rate and time, the change trends of the Hb levels, and the target Hb maintenance ratio. Adverse events (AEs) were observed and compared, and the efficacy and safety were analyzed between the two treatment groups. Additionally, the frequencies of dose adjustments between the darbepoetin alfa and epoetin alfa groups were compared during the treatment period. SAS? software version 9.2 was used to perform all statistical analyses. Descriptive statistics were used for all efficacy, safety, and demographic variable analyses, including for the primary efficacy indicators. Results::The mean Hb level was 11.3 g/dL in the darbepoetin alfa group and 10.7 g/dL in the epoetin alfa group, respectively; the difference of the lower limits of the 95% confidence intervals (CI) between the two groups was 0.1 g/dL (>-1.0 g/dL), and non-inferiority was proven; the Hb levels started to increase in the first four weeks at a similar increase rate; no obvious differences were observed between the groups in the target Hb achievement cumulative rates, and the Hb levels as well as the target Hb level maintenance rate changed over time. The incidence of AEs was 62.5% in the darbepoetin alfa group and 76.9% in the epoetin alfa group. All the adverse events observed in the study were those commonly associated with hemodialysis.Conclusion::Darbepoetin alfa intravenously once per week can effectively increase Hb levels and maintain the target Hb levels well, which makes it not inferior to epoetin alfa intravenously twice or three times per week. Darbepoetin alfa shows an efficacy and safety comparable to epoetin alfa for the treatment of renal anemia.

BACKGROUND: Unicompartmental knee arthroplasty (UKA) began in the early 1970s, which was once negated due to high postoperative repair rate. The methods for early unicompartment osteoarthritis of the knee include UKA, total knee arthroplasty, high tibial osteotomy and fibular truncation. A large number of retrospective studies and literatures have pointed that UKA is effective for unicompartment osteoarthritis of the knee and holds unique advantages.OBJECTIVE: To review the status and research progress of UKA for anteromedial compartment osteoarthritis of the knee in view of indications, contraindications, operation curve, operation process and main technical points and clinical efficacy.METHODS: The first author retrieved the databases of PubMed and CNKI from March 2006 to September 2016 using the keywords of unicompartmental knee arthroplasty, knee, osteoarthritis, clinical application in English and Chinese,respectively. A total of 95 literatures were searched, and 40 eligible articles were included in accordance with the inclusion criteria.RESULTS AND CONCLUSION: Only unilateral lesion (the degenerative cartilage surface of tibiofemoral joint) is replaced in UKA to treat early unicompartment osteoarthritis of the knee, which exhibits less trauma, rapid recovery, few complications, and normal postoperative proprioception of joint and high patient acceptance. Because of its narrow surgical indications, UKA application has been restricted compared with total knee arthroplasty. Indeed, total knee arthroplasty is matureand, and its long-term curative effect is clear. In contrast, UKA is carried out late in China, has not been popularized, and the long-term clinical efficacy remains to be verified further. But if we can accurately grasp the operation indications, choose appropriate patients, make careful preoperative preparation, and master mature skills, the clinical effect will be satisfactory. With the development of prosthesis, equipment design and operation technology, UKA will be prevailed in the treatment of anteromedial compartment osteoarthritis of the knee.

Objective To investigate the effect of high-dose creatine phosphate (CP) on myocardial ischemia-reperfusion (I/R) injury in patients undergoing cardiac valve replacement. Methods Two hundred and forty-six ASA Ⅱ or Ⅲ patients aged 42-71 yr weighing 45-80 kg undergoing mitral-aortic valve replacement were randomly assigned into 2 groups: control group ( n = 122) and CP group ( n = 124). CP 10 g in 100 ml normal saline (NS) was infused over 60 min starting from the beginning of operation in group CP. In control group NS 100 ml was infused instead of CP. Blood samples were collected before anesthesia and on 1st and 5th postoperative day for determination of serum CK, CK-MB and LDH activity and cTnI concentration. The number of patients receiving dopamine and adrenaline at the time of return of spontaneous heart beat and 12, 24 and 48 h after operation was recorded. The incidence of arrhythmia (auricular fibrillation, ventricular arrhythmia), myocardial infarct and left ventricular ejection fraction (LVEF) were also compared between the 2 groups. Results Serum CK, CK-MB and LDH activity and cTnI concentration on 1st and 5th postoperative day, the doses of dopamine and adrenalin, the incidence of arrhythmia and myocardial infarct were significantly lower in group CP than in control group. The postoperative LVEF and the incidence of spontaneous recovery of spontaneous heart-beat were significantly higher in group CP than in control group. Conclusion Pretreatment with high dose CP can protect myocardium against I/R injury in patients undergoing cardiac valve replacement under CPB.