Human keratinocyte growth factor-1 (hKGF-1), a member of the fibroblast growth factor (FGF) family, plays crucial roles in organ development, cell proliferation, wound healing, and tissue repair, representing one of the most effective and specific growth factors for skin repair. However, obtaining recombinant hKGF-1 remains challenging due to its universally low expression efficiency in vitro. This study employs the Bombyx mori baculovirus expression system to establish a technological platform that utilizes the economically important insect Bombyx mori as a bioreactor for high-efficiency and low-cost expression and production of recombinant human keratinocyte growth factor 1 (hKGF-1) protein, ultimately achieving high-level expression of hKGF-1 in Bombyx mori ovary cell line (BmN). In this study, we optimized the hKGF-1 sequence based on the codon preference of baculovirus. By fusing hKGF-1 with polyhedrin (highly expressed in this system) and adding extra promoters and enhancers, we significantly improved the expreesion level of hKGF-1 in Bombyx mori cells. The results demonstrated that the aforementioned strategies significantly enhanced the expression level of hKGF-1 in Bombyx mori cells. SDS-PAGE and Western blotting results revealed that the highest hKGF-1 expression (accounting for 8.7% of total cellular protein) was achieved when the Polh promoter was combined in tandem with the P6.9 promoter and hKGF-1 was fused with a 15-residue polyhedrin fragment for co-expression. The optimal harvest time was determined to be 120 h post transfection. This study achieved the efficient expression of hKGF-1 in Bombyx mori cells, establishing an ideal technological platform for the industrial utilization of recombinant hKGF-1. The developed methodology not only provides valuable technical references for the production of other growth factors and complex proteins, but also demonstrates significant implications for employing silkworms as bioreactors for recombinant human protein expression.
Bombyx/metabolism* ; Animals ; Baculoviridae/metabolism* ; Humans ; Fibroblast Growth Factor 7/biosynthesis* ; Recombinant Proteins/genetics* ; Cell Line ; Genetic Vectors/genetics*

Objective:To investigate the metabolic pathways and potential molecular targets associated with dapagliflozin in the treatment of type 2 diabetes mellitus.Methods:Plasma samples from patients with type 2 diabetes mellitus were collected before and after 12 months of dapagliflozin treatment and analyzed using UPLC-VION IMS Q-Tof-based metabolomics and timsTOF Pro2 diaPASEF-based proteomics. Multivariate statistical analyses were performed to identify significant differences pre- and post-treatment. Correlation analysis was then conducted to assess relationships between differentially expressed metabolites and proteins closely associated with type 2 diabetes mellitus. Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analyses were used to construct metabolic pathway maps and predict therapeutic targets.Results:After 12 months of dapagliflozin treatment, 162 differential metabolites were identified, with 59 upregulated and 103 downregulated. A total of 440 differentially expressed proteins were detected, of which 272 were upregulated and 168 were downregulated. The main classes of differential metabolites included sphingolipids, glycerophospholipids, and glycosphingolipids. Key differentially expressed proteins included importin subunit alpha-11, synemin, Janus kinase 1, and far upstream element-binding protein 2. Correlation analysis revealed 98 shared enriched pathways between differential metabolites and proteins, involving neurotrophin signaling, chemokine signaling, and B cell receptor signaling pathways. Metabolic pathway analysis suggested that dapagliflozin might regulate insulin secretion by modulating glucose-dependent insulinotropic polypeptide, calmodulin-dependent protein kinase, and diacylglycerol levels.Conclusion:Dapagliflozin may exert therapeutic effects in type 2 diabetes mellitus through multiple mechanisms, including the modulation of metabolic and proteomic profiles, participation in key cellular signaling pathways, and regulation of insulin secretion.

Oxidative stress due to aberrant metabolism is considered as a crucial contributor to diabetes and its complications. Hyperglycemia and hyperlipemia boost excessive reactive oxygen species generation by elevated mitochondrial respiration, increased nicotinamide adenine dinucleotide phosphate oxidase activity, and enhanced pro-oxidative processes, including protein kinase C pathways, hexosamine, polyol, and advanced glycation endproducts, which exacerbate oxidative stress. Oxidative stress plays a significant role in the onset of diabetes and its associated complications by impairing insulin production, increasing insulin resistance, maintaining hyperglycemic memory, and inducing systemic inflammation. A more profound comprehension of the molecular processes that link oxidative stress to diabetes is crucial to new preventive and therapeutic strategies. Therefore, this review discusses the mechanisms underlying how oxidative stress contributes to diabetes mellitus and its complications. We also summarize the current approaches for prevention and treatment by targeting the oxidative stress pathways in diabetes.
Oxidative Stress/physiology* ; Humans ; Diabetes Mellitus/physiopathology* ; Diabetes Complications/metabolism* ; Reactive Oxygen Species/metabolism* ; Glycation End Products, Advanced/metabolism* ; Animals

Objective　This study aims to investigate and compare the clinical characteristics and prognostic factors among primary liver cancer (PLC) patients who had hepatitis B virus (HBV)-induced cirrhosis associated with liver cancer, alcoholic cirrhosis associated with liver cancer, or both HBV and alcoholic cirrhosis associated with liver cancer. 　　　Methods　Inpatients diagnosed with PLC admitted to the First Affiliated Hospital of Fujian Medical University between January 2010 and September 2020 were enrolled and divided into three groups based on the etiology. The follow-up period ends in October 2024. Survival analyses were performed using Kaplan-Meier curves, univariate analysis, and multivariate Cox regression. Results　During the study period, 45 cases of alcoholic cirrhosis associated liver cancer (ALD group), and 71 cases of hepatitis B combined with alcoholic cirrhosis associated liver cancer (HBV+ALD group) were enrolled. At the same time, 73 patients with hepatitis B cirrhosis associated liver cancer (HBV group) during the same period were randomly selected with a ratio of about 1∶1.5, totaling 189 cases. And 183 (96.8%) of the patients were male and 6 (3.2%) were female. The age was (55.93±10.20) years. 109 deaths (57.7%) were recorded. The median survival times were 12 months for the entire cohort, 55 months for HBV group, 36 months for ALD group and 11 months for HBV+ALD group. And the 10-year death rate was 42.5% in HBV group, compared to 66.7% in ALD group and 67.6% in HBV+ALD group. In this study, 93 patients chose either the surgical resection or the radiofrequency ablation as their treatments. The recurrence rate was 69.9%, the median recurrence time was 8 months and the median overall survival time was 39 months. Univariate Cox regression identified that etiology of HBV and ALD, alpha-fetoprotein (AFP)>1 200 ng/mL, Child-Pugh class B and C, Barcelona Clinic Liver Cancer (BCLC) stages of C and D and curative therapies such as surgery and radiofrequency ablation were significantly correlated with overall survival (all P<0.05). Multivariate Cox regression revealed that patients with both HBV and ALD (HR=1.750，95%CI: 1.107-2.765，P=0.017), AFP>1 200 ng/mL (HR=1.649，95%CI: 1.060-2.564，P=0.027), and BCLC stages of C and D (HR=3.404，95%CI: 2.254-5.142，P<0.001) were independent risk factors of mortality in PLC patients with cirrhosis. Conclusions　Among HBV, ALD and HBV+ALD groups, the HBV+ALD group had the shortest median survival time and the highest overall mortality rate, suggesting that alcohol consumption and HBV infection may accelerate the progression of PLC with cirrhosis and worsen its prognosis. HBV infection combined with alcoholic consumption, AFP>1 200 ng/mL, and BCLC stages of C and D were independent risk factors for mortality in PLC patients with cirrhosis.

Objective·To compare the safety and short-term outcomes of robot-assisted versus laparoscopic-assisted proximal gastrectomy combined with double-flap esophagogastrostomy in the treatment of early upper gastric cancer.Methods·A retrospective cohort study was conducted to analyze the clinical and pathological data of 31 early gastric cancer patients who underwent proximal gastrectomy combined with double-flap esophagogastrostomy for gastrointestinal reconstruction at the Department of Gastrointestinal Surgery,Renji Hospital,Shanghai Jiao Tong University School of Medicine,from September 2023 to March 2024.Based on the surgical approach,patients were divided into the robot-assisted surgery group(robotic group,20 cases)and the laparoscope-assisted surgery group(laparoscopic group,11 cases).General clinical data,intraoperative conditions,and postoperative recovery between the two groups were compared.At the 6-month postoperative follow-up,upper gastrointestinal radiography and esophagogastroscopy were performed to assess anastomotic stricture and gastroesophageal reflux disease.Additionally,the gastric cancer-specific module of the European Organization for Research and Treatment of Cancer(EORTC),Quality of Life Questionnaire-Stomach 22(QLQ-STO22),was used to evaluate the patients' quality of life.Results·The general data of the two groups,including gender,age,preoperative comorbidities,American Society of Anesthesiologists(ASA)classification,Siewert classification,and pathological staging of tumors,showed no statistically significant differences(all P>0.05).All patients successfully underwent the procedure without conversion to open surgery.The time for gastroesophageal anastomosis was significantly shorter in the robotic group compared to the laparoscopic group[(31.09±8.23)min vs(43.73±8.83)min,P<0.001],while there were no statistically significant differences in other intraoperative and postoperative parameters,including operative time,intraoperative blood loss,number of lymph nodes removed,duration of gastric tube placement,time to start a liquid diet,length of postoperative hospital stay,and incidence of postoperative complications(all P>0.05).At the 6-month postoperative follow-up,30 patients completed the follow-up,with one patient lost to follow-up in the robotic group.Upper gastrointestinal radiography and esophagogastroscopy results showed that only one patient in the laparoscopic group developed an anastomotic stricture,while one patient in the robotic group developed grade A and one developed grade B gastroesophageal reflux disease(GERD).In addition,one patient in the laparoscopic group also developed grade B GERD.The incidences of GERD and anastomotic stricture showed no statistically significant differences between the two groups(both P>0.05).EORTC QLQ-STO22 results indicated that the robotic group had significantly lower scores in the dimensions of dysphagia,gastroesophageal reflux,and dietary restrictions,as well as in the total score,compared to the laparoscopic group(all P<0.05).Conclusion·Robot-assisted proximal gastrectomy combined with double-flap esophagogastrostomy is safe and feasible.It shortens anastomosis time and offers potential advantages in postoperative functional recovery and quality of life improvement.

Objective·To compare the safety and short-term outcomes of robot-assisted versus laparoscopic-assisted proximal gastrectomy combined with double-flap esophagogastrostomy in the treatment of early upper gastric cancer.Methods·A retrospective cohort study was conducted to analyze the clinical and pathological data of 31 early gastric cancer patients who underwent proximal gastrectomy combined with double-flap esophagogastrostomy for gastrointestinal reconstruction at the Department of Gastrointestinal Surgery,Renji Hospital,Shanghai Jiao Tong University School of Medicine,from September 2023 to March 2024.Based on the surgical approach,patients were divided into the robot-assisted surgery group(robotic group,20 cases)and the laparoscope-assisted surgery group(laparoscopic group,11 cases).General clinical data,intraoperative conditions,and postoperative recovery between the two groups were compared.At the 6-month postoperative follow-up,upper gastrointestinal radiography and esophagogastroscopy were performed to assess anastomotic stricture and gastroesophageal reflux disease.Additionally,the gastric cancer-specific module of the European Organization for Research and Treatment of Cancer(EORTC),Quality of Life Questionnaire-Stomach 22(QLQ-STO22),was used to evaluate the patients' quality of life.Results·The general data of the two groups,including gender,age,preoperative comorbidities,American Society of Anesthesiologists(ASA)classification,Siewert classification,and pathological staging of tumors,showed no statistically significant differences(all P>0.05).All patients successfully underwent the procedure without conversion to open surgery.The time for gastroesophageal anastomosis was significantly shorter in the robotic group compared to the laparoscopic group[(31.09±8.23)min vs(43.73±8.83)min,P<0.001],while there were no statistically significant differences in other intraoperative and postoperative parameters,including operative time,intraoperative blood loss,number of lymph nodes removed,duration of gastric tube placement,time to start a liquid diet,length of postoperative hospital stay,and incidence of postoperative complications(all P>0.05).At the 6-month postoperative follow-up,30 patients completed the follow-up,with one patient lost to follow-up in the robotic group.Upper gastrointestinal radiography and esophagogastroscopy results showed that only one patient in the laparoscopic group developed an anastomotic stricture,while one patient in the robotic group developed grade A and one developed grade B gastroesophageal reflux disease(GERD).In addition,one patient in the laparoscopic group also developed grade B GERD.The incidences of GERD and anastomotic stricture showed no statistically significant differences between the two groups(both P>0.05).EORTC QLQ-STO22 results indicated that the robotic group had significantly lower scores in the dimensions of dysphagia,gastroesophageal reflux,and dietary restrictions,as well as in the total score,compared to the laparoscopic group(all P<0.05).Conclusion·Robot-assisted proximal gastrectomy combined with double-flap esophagogastrostomy is safe and feasible.It shortens anastomosis time and offers potential advantages in postoperative functional recovery and quality of life improvement.

Objective:To investigate the metabolic pathways and potential molecular targets associated with dapagliflozin in the treatment of type 2 diabetes mellitus.Methods:Plasma samples from patients with type 2 diabetes mellitus were collected before and after 12 months of dapagliflozin treatment and analyzed using UPLC-VION IMS Q-Tof-based metabolomics and timsTOF Pro2 diaPASEF-based proteomics. Multivariate statistical analyses were performed to identify significant differences pre- and post-treatment. Correlation analysis was then conducted to assess relationships between differentially expressed metabolites and proteins closely associated with type 2 diabetes mellitus. Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analyses were used to construct metabolic pathway maps and predict therapeutic targets.Results:After 12 months of dapagliflozin treatment, 162 differential metabolites were identified, with 59 upregulated and 103 downregulated. A total of 440 differentially expressed proteins were detected, of which 272 were upregulated and 168 were downregulated. The main classes of differential metabolites included sphingolipids, glycerophospholipids, and glycosphingolipids. Key differentially expressed proteins included importin subunit alpha-11, synemin, Janus kinase 1, and far upstream element-binding protein 2. Correlation analysis revealed 98 shared enriched pathways between differential metabolites and proteins, involving neurotrophin signaling, chemokine signaling, and B cell receptor signaling pathways. Metabolic pathway analysis suggested that dapagliflozin might regulate insulin secretion by modulating glucose-dependent insulinotropic polypeptide, calmodulin-dependent protein kinase, and diacylglycerol levels.Conclusion:Dapagliflozin may exert therapeutic effects in type 2 diabetes mellitus through multiple mechanisms, including the modulation of metabolic and proteomic profiles, participation in key cellular signaling pathways, and regulation of insulin secretion.

Prior to clinical application,reliability of percutaneous ventricular assist devices(pVAD)requires to be tested systematically.Currently,there's a lack of dedicated reliability testing equipment and methodologies for pVAD.Considering the structural and functional aspects of percutaneous ventricular assist devices,this study conducts research on pVAD reliability test engineering.Test setups,clinical conditions,failure modes,effects analysis,and evaluation models have been investigated.A highly feasible methodological approach for percutaneous ventricular assist device reliability assessment has been formed.This study offers valuable insights into standardizing their reliability evaluation in clinical settings.

Objective To investigate the association between milk fat globule-epidermal growth factor 8(MFG-E8)and Alzheimer disease(AD).Methods The Alzheimer's Disease Neuroimaging Initiative database was used,and a multiple linear regression analysis was used to investigate the association of MFG-E8 with the pathological markers of AD and brain structure and perform subgroup analyses.The mixed-effects model was used to investigate the longitudinal changes in MFG-E8 with AD pathological markers and brain structure,and a mediation analysis was used to investigate the potential association of MFG-E8 with AD pathology and brain structure.Results A total of 377 individuals were en-rolled in the study.The level of MFG-E8 was positively correlated with the concentrations of amyloid β-protein 42(Aβ42),tau protein,and phosphorylated tau protein in cerebrospinal fluid,and subgroup analyses showed that the correlation of MFG-E8 with the concentrations of AD pathological markers and lateral ventricular volume in male individuals,the elderly popula-tion,and the individuals carrying the APOE4 gene was consistent with that in the overall population.Longitudinal studies showed that during follow-up,the increase in MFG-E8 level was significantly associated with enlarged lateral ventricle volume.In addition,MFG-E8 could alter lateral ventricle volume by affecting the level of Aβ42.Conclusion MFG-E8 is associated with AD and may influence lateral ventricular volume through AD pathological markers.

Objective:To summarize the clinical characteristics in of different antibody subtypes in of patients with antisynthetase syndrome (ASS) complicated with interstitial lung disease (ILD).Methods:A retrospective analysis was conducted on 132 ASS-ILD patients at the First Affiliated Hospital of Nanjing Medical University (Jiangsu Provincial People′s Hospital), encompassing a period from December 2019 to June 2023. The data included were basic demographic information, clinical features, laboratory test results, chest computed tomography (CT) scans, and pulmonary lung function tests. Patients were categorized into distinct subtypes based on anti-aminoacyl tRNA synthetase (ARS) antibodies. Statistical analysis was performed using a t-test for comparing means between two samples with equal variance, the Mann-Whitney U test for non-normally distributed continuous data, and the chi-square ( χ2) test or Fisher′s exact test for categorical variables. Results:The most prevalent subtype of anti-synthetase antibody was anti-histidine antibody (Jo-1), accounting for 60 of 132 cases (45.5%), followed by anti-glycine-based tRNA synthetase antibody (EJ) (33/132, 2 5.0%), anti-tRNA synthase antibody (PL-7) (26/132, 19.7%), anti-alanine-based tRNA synthetase antibody (PL-12) (7.6%, 10/132), anti-isoleucine-tRNA synthase antibody (OJ) (3/132, 2.2%). The presence of anti-Ro-52 antibodies was significantly associated with rapidly progressive ILD. In patients with different subtypes of ASS-ILD, the presence of anti-Jo-1 antibodies is was positive in 28 cases (46.7%), and the combination of infection is was more common than in other groups ( χ2=0.15, P=0.047). The group with positive anti-EJ antibodies has had a significant decline in lung function, and cough is was more common in 31 cases (93.9%) than in other groups ( P<0.05); the group with positive anti-PL-12 antibodies has had a more pronounced decline in lung function than other groups ( P<0.05), and fever (7 cases, 70.0%) wais more common than in other groups ( χ2=0.02, P=0.022). Conclusion:Anti-Jo-1, Anti-PL-7, and Anti-PL-12 antibodies were are observed more frequently in patients with ILD. Furthermore, a significant deterioration in lung function was is observed in patients testing positive for anti-PL-12 and anti-EJ antibodies.