OBJECTIVE: To analyze the effectiveness of single three-dimensional (3D)-printed microporous titanium prostheses and flap combined prostheses implantation in the treatment of large segmental infectious bone defects in limbs. METHODS: A retrospective analysis was conducted on the clinical data of 76 patients with large segmental infectious bone defects in limbs who were treated between January 2019 and February 2024 and met the selection criteria. Among them, 51 were male and 25 were female, with an age of (47.7±9.4) years. Of the 76 patients, 51 had no soft tissue defects (single prostheses group), while 25 had associated soft tissue defects (flap combined group). The single prostheses group included 28 cases of tibial bone defects, 11 cases of femoral defects, 5 cases of humeral defects, 4 cases of radial bone defects, and 3 cases of metacarpal, or carpal bone defects, with bone defect length ranging from 3.5 to 28.0 cm. The flap combined group included 3 cases of extensive dorsum of foot soft tissue defects combined with large segmental metatarsal bone defects, 19 cases of lower leg soft tissue defects combined with large segmental tibial bone defects, and 3 cases of hand and forearm soft tissue defects combined with metacarpal, carpal, or radial bone defects, with bone defect length ranging from 3.8 to 32.0 cm and soft tissue defect areas ranging from 8 cm×5 cm to 33 cm×10 cm. In the first stage, vancomycin-loaded bone cement was used to control infection, and flap repair was performed in the flap combined group. In the second stage, 3D-printed microporous titanium prostheses were implanted. Postoperative assessments were performed to evaluate infection control and bone integration, and pain release was evaluated using the visual analogue scale (VAS) score. RESULTS: All patients were followed up postoperatively, with an average follow-up time of (35.2±13.4) months. In the 61 lower limb injury patients, the time of standing, walk with crutches, and fully bear weight were (2.2±0.6), (3.9±1.1), and (5.4±1.1) months, respectively. The VAS score at 1 year postoperatively was significantly lower than preoperative one ( t=-10.678, P<0.001). At 1 year postoperatively, 69 patients (90.8%) showed no complication such as infection, fracture, prosthesis displacement, or breakage, and X-ray films indicated good integration at the prosthesis-bone interface. According to the Paley scoring system for the healing of infectious bone defects, the results were excellent in 37 cases, good in 29 cases, fair in 3 cases, and poor in 7 cases. In the single prostheses group, during the follow-up, there was 1 case each of femoral prostheses fracture, femoral infection, and tibial infection, with a treatment success rate of 94.1% (48/51). In lower limb injury patients, the time of fully bear weight was (5.0±1.0) months. In the flap combined group, during the follow-up, 1 case of tibial fixation prostheses screw fracture occurred, along with 2 cases of recurrent foot infection in diabetic patients and 1 case of tibial infection. The treatment success rate was 84.0% (21/25). The time of fully bear weight in lower limb injury patients was (5.8±1.2) months. The overall infection eradication rate for all patients was 93.4% (71/76). CONCLUSION The use of 3D-printed microporous titanium prostheses, either alone or in combination with flaps, for the treatment of large segmental infectious bone defects in the limbs results in good effectiveness with a low incidence of complications. It is a feasible strategy for the reconstruction of infectious bone defects.
Humans ; Male ; Female ; Middle Aged ; Printing, Three-Dimensional ; Titanium ; Retrospective Studies ; Surgical Flaps ; Adult ; Prosthesis Implantation/methods* ; Plastic Surgery Procedures/methods* ; Treatment Outcome ; Prostheses and Implants ; Bone Diseases, Infectious/surgery* ; Extremities/surgery* ; Prosthesis Design

OBJECTIVE To investigate the stability of 5-fluorouracil （5-FU） in human blood and to establish a standardized clinical sampling and delivery procedure for therapeutic drug monitoring （TDM） of 5-FU. METHODS The EDTA-anticoagulated whole blood was used as the matrix to prepare stability assessment samples of 5-FU at both low （200 ng/mL） and high （5 000 ng/mL） concentrations （with groups without stabilizer and with 1% volume ratio of stabilizer）. The stability assessment samples were placed under room temperature （[ 25±2） ℃] and refrigerated （2-8 ℃） conditions， with sampling at 0， 0.5， 1， 2， 4， 7， and 24 h. After vortexing and centrifugation， the upper plasma layer was collected； proteins were precipitated using methanol， and the concentration of 5-FU in plasma was determined by liquid chromatography-tandem mass spectrometry. Based on the whole blood stability results， clinical sampling and delivery procedures were established. RESULTS The concentration of 5-FU in blank whole blood samples without stabilizers was significantly lower than that in samples with stabilizers （P＜0.05）. However， varying volumes （10， 25， 50 μL） of stabilizers had no significant effect on the measured concentrations of 5-FU in stability assessment samples with low and high concentrations （P＞0.05）. Without the addition of a stabilizer， low- and high-concentration 5-FU whole blood samples remained stable at room temperature for 0.5 h and 1 h， respectively， and under refrigeration for 2 h and 7 h， respectively. After the addition of a 1% stabilizer， the whole blood samples remained stable for up to 24 h under both room temperature and refrigerated conditions. Based on these findings， the following procedure was established： after collection， whole blood samples could be temporarily stored at room temperature （≤0.5 h） or at 4　℃ （≤2 h）， and transported at 2-8　℃. Upon delivery to the laboratory， a 1% volume ratio of stabilizer must be added immediately， followed by centrifugation within 24 h. The resulting plasma should be stored at －20 ℃ . CONCLUSIONS 5-FU in whole blood exhibits poor stability at room temperature. Refrigeration at 2-8　℃ slightly improves stability ， but degradation still occurs rapidly. Adding a stabilizer at a 1% volume ratio significantly prolongs the refrigerated storage time. The established sampling and transport procedure for 5-FU TDM innovatively introduces the stabilizer addition step at the laboratory sample reception stage （rather than immediately after blood draw）. This approach ensures analytical quality while offering greater adaptability to real-world clinical sampling conditions， significantly improving practical feasibility.

Loss-of-function variants in CSDE1 have been strongly linked to neuropsychiatric disorders, yet the precise role of CSDE1 in neurogenesis remains elusive. In this study, we demonstrate that knockout of Csde1 during cortical development in mice results in impaired neural progenitor proliferation, leading to abnormal cortical lamination and embryonic lethality. Transcriptomic analysis revealed that Csde1 upregulates the transcription of genes involved in the cell cycle network. Applying a dual thymidine-labelling approach, we further revealed prolonged cell cycle durations of neuronal progenitors in Csde1-knockout mice, with a notable extension of the G1 phase. Intersection with CLIP-seq data demonstrated that Csde1 binds to the 3' untranslated region (UTR) of mRNA transcripts encoding cell cycle genes. Particularly, we uncovered that Csde1 directly binds to the 3' UTR of mRNA transcripts encoding Cdk6, a pivotal gene in regulating the transition from the G1 to S phases of the cell cycle, thereby maintaining its stability. Collectively, this study elucidates Csde1 as a novel regulator of Cdk6, sheds new light on its critical roles in orchestrating brain development, and underscores how mutations in Csde1 may contribute to the pathogenesis of neuropsychiatric disorders.
Animals ; Neurogenesis/genetics* ; Cell Cycle/genetics* ; Mice, Knockout ; Mice ; Neural Stem Cells/metabolism* ; DNA-Binding Proteins/metabolism* ; Cyclin-Dependent Kinase 6/genetics* ; Cell Proliferation ; 3' Untranslated Regions ; Cerebral Cortex/embryology* ; RNA-Binding Proteins ; Mice, Inbred C57BL

Non-small cell lung cancer (NSCLC) ranks the first among malignant tumors in China and even in the whole world. In recent years, the developement of genetic testing technology, particularly tumor gene sequencing, has provided a solid basis for the clinical molecular diagnosis of NSCLC, which has greatly increased the chances of patients benefiting from targeted therapy or immunotherapy, and ultimately extending their survival. Standardizing the use of tumor gene sequencing is crucial for the precision medicine in NSCLC. This paper discusses the normalization of tumor gene sequencing technology in the clinical molecular diagnostic pathway of NSCLC, which may promote the standardized use of tumor gene sequencing technology in targeted therapy or immunotherapy drug selection, toxicity and side effect prediction, efficacy monitoring, recurrence and prognosis evaluation of NSCLC patients. This article discusses the standardization of the application of tumor gene sequencing technology in the clinical molecular diagnosis pathway of NSCLC. Additionally, it offers a foundation for the uniform use of tumor gene sequencing technology in other solid tumors.

Objective To explore the effect of nicotinamide transmethylase on intracellular reactive oxygen species(ROS)in iron death of hepatocellular carcinoma cells and its mechanism.Methods Methyl nicotinamide(MNA)expression in cells was detected using a tandem liquid chromatography-mass spectrometry.The average fluorescence intensity of ROS and lipid peroxidation was measured using a flow cytometer.Western blot was used to detect changes in the expression of human liver cancer cells(SK-Hep-1,Hep3B).Forty patients with primary hepatocellular carcinoma who received treatment in our hospital from March 2019 to February 2020 were selected as the study subjects,and their adjacent tissue samples and liver cancer tissue samples were collected.Immunohistochemical methods were used to detect the levels of nicotinamide N-methyltransferase(NNMT)and ROS in adjacent and liver cancer tissues.CCK-8 method was used to detect the survival activity of cells with different iron concentrations.Results The MNA levels in the liver cancer tissue group were higher than those in the adjacent tissue group(P<0.05).Compared with the adjacent tissue group,the average fluorescence intensity expression of ROS in the liver cancer tissue group increased,while the average fluorescence intensity expression of lipid peroxidation decreased(P<0.05).Compared with the adjacent tissue group,the expression levels of SK Hep-1 and Hep3B cells in the liver cancer tissue group increased(P<0.05).Compared with the control group,NNMT groups 2,10,20,and 25 μmol/L The cell survival activity level increased(P<0.05);Compared with the NNMT group,the iron inhibition group had different iron concentrations(2μmol/L,10μmol/L,20μmol/L,25μmol/L.The expression of cell viability decreased(P<0.05).Conclusion ROS mediated by nicotinamide methyltransferase can be guided to produce ROS and energy disorders,leading to increased tumor cell death.

Diabetic cardiomyopathy(DCM)is a unique myocardial disease caused by diabetes mellitus,which can increase the risk of heart failure and death,and is one of the main causes of death of diabetes mellitus patients worldwide.Although the research on the pathogenesis of DCM has made great progress,it has not yet been fully clarified.Many studies have shown that long noncoding RNAs(lncRNAs)can interact with microRNAs(miRNAs)as competitive endogenous RNAs(ceRNAs),participate in the regulation of gene expression,and then affect the development of DCM.This article gives an overview of lncRNAs and its biological functions as well as ceRNA hypothesis,and focuses on the role of lncRNAs as ceRNAs in regulating the occurrence and development of DCM.

Objective:To systematically evaluate the psychological experience of financial toxicity in breast cancer patients.Methods:A computer search was conducted in Web of Science, PubMed, Cochrane Library, Embase, CINAHL, China National Knowledge Infrastructure, China Biology Medicine disc, Wanfang, and VIP for qualitative studies on the psychological experience of financial toxicity among breast cancer patients up to April 15, 2023. The Joanna Briggs Institute's quality assessment criteria for qualitative research were used for literature quality evaluation, and an aggregative integration method was applied for data analysis.Results:Ten studies were included, from which 56 distinct themes were extracted. These themes were consolidated into 11 new categories, forming three integrated results: multidimensional negative experiences in coping with financial toxicity, needs and expectations in dealing with financial toxicity, and strategies for dealing with financial toxicity.Conclusions:Breast cancer patients face varying degrees of financial toxicity, negatively impacting their physical and mental health. Healthcare professionals should pay close attention to the characteristics and needs of patients coping with financial toxicity. Continuous assessment of their financial status and implementation of comprehensive intervention strategies and measures through multiple channels and approaches are needed to help reduce the issues of financial toxicity.

AIM:To investigate the pharmacoki-netic/pharmacodynamic(PK/PD)profile of esome-prazole for injection in critically ill patients.METH-ODS:This was a prospective,single-center,open-la-bel study,all patients received intravenous infused esomeprazole 40 mg q12h for stress ulcer prophy-laxis,treatment duration is determined by clini-cians based on patients condition.Forty critically ill patients were enrolled and were divided into single and multiple dose groups according to the timing of blood sample collection.Twenty-one patients in the single-dose group had their blood samples col-lected at 1,3,6,8,and 12 h after the first dose,and 34 patients in the multiple-dose group had their blood samples collected at 0 h before the fifth dose and 1,3,6,and 8,and 12 h after the fifth dose,of which 14 patients had their blood samples collected at both the first dose and the repeated doses.The concentration of esomeprazole was measured by HPLC-MS/MS,and PK parameters were analyzed using noncompartmental analysis.Gastric aspirates were collected for pH measure-ment in fasted patients with gastric tube before the first dose(0 h),and 1,2,4,8,12,14,16,20,24 h after the initiation of drug administration,and the percentage of time with pH≥4 was calculated.All adverse events and serious adverse events during treatment were recorded.RESULTS:Patients in the single-dose group were 67.75 years old(45-69 years)with a BMI(24.05±3.35)kg/m2,and pa-tients in the multiple-dose group were 63.35 years old(24-87 years)with a BMI(24.08±3.29)kg/m2.PK parameters after the first dose were AUC0-t(11.26±6.58)mg·h·L-1,Cmax(3.08±2.06)mg/L,CL(4.13±3.68)L/h,Vd(17.12±6.13)L,t1/2(4.80±3.06)h;PK parameters after multiple doses were AUC0-t(16.70±11.20)mg·h/L,Cmax(3.37±2.59)mg/L,CL(3.94±2.94)L/h,Vd(22.71±17.26)L,t1/2(5.23±3.34)h.Percentage of time with pH≥4 with-in 0 h-24 h after administration was 61.69％,and percentage of time with pH≥4 within 12 h-24 h was up to 100％.Esomeprazole was well tolerated by all patients with no serious adverse events.CONCLU-SION:Compared with healthy volunteers,inject-able esomeprazole showed increased Vd,decreased CL,increased drug exposure and accumulation af-ter repeated administration in critically ill patients.The drug had a favorable safety profile in critically ill patients.

Objective To establish a palmitic acid(PA)-induced model of lipotoxic bone formation inhibition in zebrafish.Methods AB strain zebrafish embryos were divided randomly into a blank control group,PA group,and simvastatin(SIM)group.Embryos in the PA and SIM groups received PA from 3 days post-fertilization(dpf),and embryos in the SIM group received SIM continuously for 4 days from 5 dpf.Establishment of the model was confirmed at 9 dpf by calcein staining,Nile red staining,triglyceride and total cholesterol content determination,and q-PCR.Results PA significantly decreased the number of vertebrae,promoted lipid accumulation,increased triglyceride and total cholesterol contents,promoted the expression of lipid-related genes PPARγ、c/EBPα,and inhibited the expression of osteogenic genes ALP and RUNX2.SIM improved the inhibitory effect of PA on bone formation in zebrafish.Conclusions PA can successfully create a lipotoxic model of bone-formation inhibition,similar to the pathological process of osteoporosis,using a simple,sensitive,and controllable method.This model can then be used for drug screening for osteoporosis and related diseases.

Objective To establish a fast,stable,and sensitive zebrafish model of osteoporosis(OP)using different method.Methods OP models were induced by iron overload or prednisolone(Pred),and bone formation and mortality were observed.The groups were divided into:Control group,model group(include FAC group and Pred group),and positive control group(AC group).Ammonium ferric citrate was used as the model drug in the iron-overload induction method.For the Pred induction models,the modeling time for the Pred-3 days post-fertilization(dpf)method was 3～9 dpf,the modeling time for the Pred-5 dpf method was 5～10 dpf,and Pred was administered from 3 dpf and removed from 7～9 dpf for the Pred withdrawal method.To compare the anti-osteoporosis(OP)effects of commonly used drugs such as Alfacalcidol(AC),Calcitriol(CA),and Alendronate(AL),it's important to select a stable and sensitive positive control drug and to further optimize different staining methods and conditions.Results There was no significant effect of ammonium ferric citrate 500 μg/mL on bone formation.Bone formation and the length of the first vertebra were significantly decreased in the Pred group induced by Pred-3 dpf compared with those in the control group(P＜0.01,P＜0.05),but zebrafish mortality was higher.There was no significant difference between the Pred-5 dpf method,but bone formation was significantly reduced in the Pred withdrawal group(P＜0.01),with no mortality.Alfacalcidol,calcitriol,and alendronate all had anti-OP effects,with CA having the most sensitive and stable anti-OP effect.Alizarin red staining showed that the optimal dye parameters were 0.02%concentration for dyeing 2 h,with washing in 0.5%KOH and glycerol under the conditions of a 3∶1 ratio for 3 h followed by a 1∶1 ratio for 14 h.The result of staining showed that calcein was more sensitive for staining bone nodes and ARS staining was more sensitive for staining the first vertebra.Conclusions The Pred withdrawal method can be used to establish a rapid,stable,and sensitive OP model in zebrafish as a reliable model for studying OP.