AIM:To investigate the effect of dyes,Remazol BrOrange yellow(RBY)and erythrosine(ERY),on the outcomes of immunoblotting analysis when used for staining the concentrate gel in sodium dodecyl sulfate-polyacryl-amide gel electrophoresis(SDS-PAGE).METHODS:Polyacrylamide gels were divided into five groups:the control group(prepared according to the conventional kit protocol),the RBY-stained group with a final concentration of 0.08 g/L,the RBY-stained group with a final concentration of 0.16 g/L,the ERY-stained group with a final concentration of 0.08 g/L,and the ERY-stained group with a final concentration of 0.8 g/L.Gels were prepared and subjected to electro-phoresis,followed by coomassie brilliant blue staining to visualize protein bands.Subsequently,proteins were transferred to PVDF membranes,which were then blocked,incubated with primary and secondary antibodies,washed,and finally ex-posed for imaging to observe the target protein vinculin bands.RESULTS:Compared with the unstained concentrate gel,the loading wells of the RBY or ERY pre-stained concentrate gel were more clearly visible.Analysis of the gels stained with coomassie brilliant blue after electrophoresis and marker visualization showed no significant different in protein elec-trophoretic mobility between prestained and unstained gels.Comparative analysis of the immunoblotting also indicated that the detection of protein samples transferred to PVDF membranes was unaffected.CONCLUSION:Prestaining concen-trate gels with RBY or ERY can enhance the efficiency of gel-based electrophoresis and immunoblotting analysis.

This article is to evaluate the clinical outcomes and health economics of minimally invasive surgery for hypertensive intracerebral hemorrhage(HICH).This review systematically compares clinical efficacy and economic value of three minimally invasive techniques:small bone window microsurgery,neuroendoscopic surgery and stereotactic drainage based on 12 randomized controlled trials(RCTs),7 cohort studies,and 8 economic evaluations 2019-2024.Cost-effectiveness analysis(CEA)and cost-utility analysis(CUA)were employed to assess resource utilization and health outcomes.Minimally invasive approaches overall outperform conventional craniotomy.Stereotactic surgery achieves the shortest hospitalization(5-8 days)and lowest direct costs;neuroendoscopy significantly improves quality of life(quality-adjusted life years(QALYs);and small bone window surgery offers the best postoperative stability.It is recommended to choose the surgical method based on patient characteristics and to optimize healthcare resource allocation through medical insurance payment reform and technology promotion.

AIM:To investigate the effect of dyes,Remazol BrOrange yellow(RBY)and erythrosine(ERY),on the outcomes of immunoblotting analysis when used for staining the concentrate gel in sodium dodecyl sulfate-polyacryl-amide gel electrophoresis(SDS-PAGE).METHODS:Polyacrylamide gels were divided into five groups:the control group(prepared according to the conventional kit protocol),the RBY-stained group with a final concentration of 0.08 g/L,the RBY-stained group with a final concentration of 0.16 g/L,the ERY-stained group with a final concentration of 0.08 g/L,and the ERY-stained group with a final concentration of 0.8 g/L.Gels were prepared and subjected to electro-phoresis,followed by coomassie brilliant blue staining to visualize protein bands.Subsequently,proteins were transferred to PVDF membranes,which were then blocked,incubated with primary and secondary antibodies,washed,and finally ex-posed for imaging to observe the target protein vinculin bands.RESULTS:Compared with the unstained concentrate gel,the loading wells of the RBY or ERY pre-stained concentrate gel were more clearly visible.Analysis of the gels stained with coomassie brilliant blue after electrophoresis and marker visualization showed no significant different in protein elec-trophoretic mobility between prestained and unstained gels.Comparative analysis of the immunoblotting also indicated that the detection of protein samples transferred to PVDF membranes was unaffected.CONCLUSION:Prestaining concen-trate gels with RBY or ERY can enhance the efficiency of gel-based electrophoresis and immunoblotting analysis.

This article is to evaluate the clinical outcomes and health economics of minimally invasive surgery for hypertensive intracerebral hemorrhage(HICH).This review systematically compares clinical efficacy and economic value of three minimally invasive techniques:small bone window microsurgery,neuroendoscopic surgery and stereotactic drainage based on 12 randomized controlled trials(RCTs),7 cohort studies,and 8 economic evaluations 2019-2024.Cost-effectiveness analysis(CEA)and cost-utility analysis(CUA)were employed to assess resource utilization and health outcomes.Minimally invasive approaches overall outperform conventional craniotomy.Stereotactic surgery achieves the shortest hospitalization(5-8 days)and lowest direct costs;neuroendoscopy significantly improves quality of life(quality-adjusted life years(QALYs);and small bone window surgery offers the best postoperative stability.It is recommended to choose the surgical method based on patient characteristics and to optimize healthcare resource allocation through medical insurance payment reform and technology promotion.

Objective:To investigate the clinical characteristics and prognostic factors of malignant peritoneal mesothelioma (MPeM).Methods:A retrospective case series study was conducted. Clinical and follow-up data of 73 MPeM patients who received pemetrexed and cisplatin (AP regimen)-based treatment at the Cancer Hospital of the Chinese Academy of Medical Sciences from January 2004 to December 2022 were collected. The primary endpoints were overall survival (OS) and progression-free survival (PFS). Kaplan-Meier method was used to perform the survival analysis; univariate and multivariate analyses were performed to identify the influencing factors of prognosis by log-rank test and Cox proportional hazards model.Results:In 73 MPeM patients, there were 33 males and 40 females, with a median age of 57 years old (range: 20-76 years old). Among them, 41 patients (56.2%) were aged ≥55 years old, 5 patients (6.8%) had a history of asbestos exposure, 45 patients (61.6%) presented with ascites, and 32 patients (43.8%) had distant metastasis, 70 patients (95.9%) were epithelioid subtype, 38 patients (52.1%) underwent surgery, and 3 patients (4.1%) received radiotherapy. The median OS time of all patients was 30 months (95% CI: 25-50 months), and the median PFS time was 8 months (95% CI: 6-14 months). Univariate analysis results showed that the differences in OS and PFS between patients with different ages (<55 years old vs. ≥55 years old: the median OS time not reached vs. 25 months, P < 0.001; the median PFS time 13 months vs. 7 months, P = 0.046) and Eastern Cooperative Oncology Group (ECOG) score (1 point vs. 2 points: the median OS time 37 months vs. 21 months, P < 0.001; the median PFS time 14 months vs. 4 months, P = 0.004) were statistically significant. There were statistically significant differences in OS among patients with different status of surgery (with vs. without: the median OS time 37 months vs. 24 months, P = 0.020), history of asbestos exposure (with vs. without: the median OS time 32 months vs. 18 months, P = 0.002) and distant metastasis (with vs. without: the median OS time 58 months vs. 20 months, P < 0.001). Multivariate analysis results showed that ECOG score of 2 points ( HR = 5.04, 95% CI: 1.29-19.73, P = 0.020) and distant metastasis ( HR = 4.26, 95% CI: 1.77-10.24, P = 0.001) were independent risk factors for OS of patients. Conclusions:Most MPeM patients are female, the epithelioid subtype is predominant, and the overall prognosis is poor. However, patients aged <55 years old, without history of asbestos exposure, with a good general condition, with surgery, or without distant metastasis have relatively good prognosis.

Objective To investigate the effect of miR-2110 on the biological behaviors,such as cell proliferation,apoptosis,and metastasis,of lung adenocarcinoma(LUAD)cells by means of cell and animal experiments.Methods Bioinformatics websites,including ENCORI,TargetScan,miRTarBase,and Tarbase,were used to analyze the changes in the expression of miR-2110 in LUAD samples and to predict miR-2110 target.LUAD tissue samples and cells were collected and the changes in the expression of miR-2110 were verified through PCR technology.CCK-8 assay,clonogenic assay,Transwell assay,and flow cytometry were conducted to analyze alterations in the functions of LUAD cells.In addition,10 BALB/c female nude mice aged 6 to 8 weeks were randomly divided into 2 groups,and the effect of miR-2110 on LUAD was investigated by in vivo experiments.Results miR-2110 was significantly decreased in LUAD tissues and cells compared with the normal lung tissues.miR-2110 overexpression inhibited the proliferation and metastasis of LUAD cells and promoted the apoptosis of tumor cells(P＜0.05).Bioinformatics prediction and dual luciferase reporter gene assay results confirmed that miR-2110 could target and bind to CDT1.In addition,overexpression of CDT1 gene reversed the proliferation,metastasis,and apoptosis of miR-2110 compared with the miR-2110 overexpression group(P＜0.05).Nude mice in vivo experiments showed that miR-2110 overexpression significantly decreased the expression of Ki67,a tumor proliferation index,and vimentin and MMP9,two metastasis indices,compared with the control group.Conclusion miR-2110 can inhibit proliferation and metastasis of LUAD by targeting CDT1,providing a new rationale for the treatment of LUAD.

OBJECTIVE To confirm the therapeutic effect of Jintiange capsules on osteoarthritis(OA) and the potential mechanism of synovial mesenchymal stem cell exosomes(SMSC-Exos) and articular chondrocytes(ACs) in the treatment of OA based on high-throughput sequencing technology. METHODS Type Ⅱ collagenase-induced OA rats were used for efficacy verification through general behavioral observation, bipedal balance difference experiment, mechanical foot reflex threshold, Micro-CT observation, and Safranin O-Fast Green staining. SMSCs and ACs were cultured in suitable concentration of drug-containing serum, and mRNA sequencing was performed on ACs in the control, model, and Jintiange capsules groups, as well as miRNA sequencing on SMSC-Exos. Differential expressed mRNAs and miRNAs were screened and target genes were predicted. The common differential expressed genes between SMSC and ACs were obtained by intersecting the differential expressed genes, and a miRNA-mRNA regulatory network was constructed using Cytoscape software. The expression trend analysis of common differential expressed genes was conducted, as well as the correlation analysis between differential expressed gene mRNA and miRNA, Micro-CT efficacy indicators, and differential expressed gene mRNA. RESULTS Under the pathological state of OA, the expression of miRNA-23a-3p, miRNA-342-3p, miRNA-146b-5p, miRNA-501-3p, and miRNA-214-3p were down-regulated, while miRNA-222-3p, miRNA-30e-3p, miRNA-676-3p, and miRNA-192-5p were up-regulated (P<0.05). The expressions of these miRNAs were significantly reversed after intervention with drug-containing serum of Jintiange capsules. There was a certain correlation between Micro-CT efficacy indicators, mRNA and miRNA. CONCLUSION Jintiange capsule has obvious efficacy in the treatment of OA, and its mechanism may be related to the promotion of SMSC-Exos targeting ACs to transport miRNA and then regulate Serpinb10, Ntn1, Il1b, Tgm2, Megf10, Il11, Cd40, Slc15a3, Pou2f2 and other genes.

BACKGROUND:As tissue engineering brings new hope to the worldwide problem of articular cartilage repair,the construction of light-curing 3D printed hydrogel scaffolds with biomimetic composition is of great significance for cartilage tissue engineering. OBJECTIVE:To construct a biomimetic methacryloylated hyaluronic acid/acellular Wharton's jelly composite hydrogel scaffold by digital light processing 3D printing technology,and to evaluate its biocompatibility. METHODS:Wharton's jelly was isolated and extracted from human umbilical cord,then decellulated,freeze-dried,ground into powder,and dissolved in PBS to prepare 50 g/L acellular Wharton's jelly solution.Methylallylated hyaluronic acid was prepared,lyophilized and dissolved in PBS to prepare 50 g/L methylallylated hyaluronic acid solution.Acellular Wharton's jelly solution was mixed with methacrylyacylated hyaluronic acid solution at a volume ratio of 1:1,and was used as bio-ink after adding photoinitiator.Methylacrylylated hyaluronic acid hydrogel scaffolds(labeled as HAMA hydrogel scaffolds)and methylacrylylated hyaluronic acid/acellular Wharton's jelly gel scaffolds(labeled as HAMA/WJ hydrogel scaffolds)were prepared by digital light processing 3D printing technology,and the microstructure,swelling performance,biocompatibility,and cartilage differentiation performance of the scaffolds were characterized. RESULTS AND CONCLUSION:(1)Under scanning electron microscope,the two groups of scaffolds showed a three-dimensional network structure,and the fiber connection of HAMA/WJ hydrogel scaffold was more uniform.Both groups achieved swelling equilibrium within 10 hours,and the equilibrium swelling ratio of HAMA/WJ hydrogel scaffold was lower than that of HAMA hydrogel scaffold(P＜0.05).(2)CCK-8 assay showed that HAMA/WJ hydrogel scaffold could promote the proliferation of bone marrow mesenchymal stem cells compared with HAMA hydrogel scaffold.Dead/live staining showed that bone marrow mesenchymal stem cells grew well on the two groups of scaffolds,and the cells on the HAMA/WJ hydrogel scaffolds were evenly distributed and more cells were found.Phalloidine staining showed better adhesion and spread of bone marrow mesenchymal stem cells in HAMA/WJ hydrogel scaffold than in HAMA.(3)Bone marrow mesenchymal stem cells were inoculated into the two groups for chondrogenic induction culture.The results of qRT-PCR showed that the mRNA expressions of agglutinoglycan,SOX9 and type Ⅱ collagen in the HAMA/WJ hydrogel scaffold group were higher than those in the HAMA hydrogel scaffold group(P＜0.05,P＜0.01).(4)These findings indicate that the digital light processing 3D bioprinting HAMA/WJ hydrogel scaffold can promote the proliferation,adhesion,and chondrogenic differentiation of bone marrow mesenchymal stem cells.

Objective:To investigate the clinical characteristics and prognostic factors of malignant peritoneal mesothelioma (MPeM).Methods:A retrospective case series study was conducted. Clinical and follow-up data of 73 MPeM patients who received pemetrexed and cisplatin (AP regimen)-based treatment at the Cancer Hospital of the Chinese Academy of Medical Sciences from January 2004 to December 2022 were collected. The primary endpoints were overall survival (OS) and progression-free survival (PFS). Kaplan-Meier method was used to perform the survival analysis; univariate and multivariate analyses were performed to identify the influencing factors of prognosis by log-rank test and Cox proportional hazards model.Results:In 73 MPeM patients, there were 33 males and 40 females, with a median age of 57 years old (range: 20-76 years old). Among them, 41 patients (56.2%) were aged ≥55 years old, 5 patients (6.8%) had a history of asbestos exposure, 45 patients (61.6%) presented with ascites, and 32 patients (43.8%) had distant metastasis, 70 patients (95.9%) were epithelioid subtype, 38 patients (52.1%) underwent surgery, and 3 patients (4.1%) received radiotherapy. The median OS time of all patients was 30 months (95% CI: 25-50 months), and the median PFS time was 8 months (95% CI: 6-14 months). Univariate analysis results showed that the differences in OS and PFS between patients with different ages (<55 years old vs. ≥55 years old: the median OS time not reached vs. 25 months, P < 0.001; the median PFS time 13 months vs. 7 months, P = 0.046) and Eastern Cooperative Oncology Group (ECOG) score (1 point vs. 2 points: the median OS time 37 months vs. 21 months, P < 0.001; the median PFS time 14 months vs. 4 months, P = 0.004) were statistically significant. There were statistically significant differences in OS among patients with different status of surgery (with vs. without: the median OS time 37 months vs. 24 months, P = 0.020), history of asbestos exposure (with vs. without: the median OS time 32 months vs. 18 months, P = 0.002) and distant metastasis (with vs. without: the median OS time 58 months vs. 20 months, P < 0.001). Multivariate analysis results showed that ECOG score of 2 points ( HR = 5.04, 95% CI: 1.29-19.73, P = 0.020) and distant metastasis ( HR = 4.26, 95% CI: 1.77-10.24, P = 0.001) were independent risk factors for OS of patients. Conclusions:Most MPeM patients are female, the epithelioid subtype is predominant, and the overall prognosis is poor. However, patients aged <55 years old, without history of asbestos exposure, with a good general condition, with surgery, or without distant metastasis have relatively good prognosis.