BACKGROUND:The inflammatory response of microglia is closely related to neuronal survival,regeneration,and functional recovery after spinal cord injury.Peroxiredoxin 1 is not only involved in the regulation of oxidative stress,but also has an important effect on cell proliferation,apoptosis,and inflammatory response.OBJECTIVE:To investigate the role and mechanism of peroxiredoxin 1 in the inflammatory response of microglia following spinal cord injury.METHODS:(1)Twelve female C57BL/6 mice were randomly divided into sham-operated(n=6)and spinal cord injury(n=6)groups.The sham-operated group was not modeled and acute spinal cord injury models were constructed in the spinal cord injury group using the modified Allen's method.Spinal cord tissue at the injured site was taken at 7 days after modeling and transcriptome sequencing was performed to identify differentially expressed genes.The expression of peroxiredoxin 1 in spinal cord tissues was verified using western blot and RT-qPCR.(2)Mouse microglia BV2 were divided into two groups:the control group was stimulated with lipopolysaccharide for 6 hours,and in the knockout group,lipopolysaccharide stimulation was applied for 6 hours at 24 hours after peroxiredoxin 1 was knocked down in the cells.RT-qPCR was performed to detect mRNA expression of peroxiredoxin 1,inflammatory factors(interleukin 1β,interleukin 6,inducible nitric oxide synthase,tumor necrosis factor α,C-C motif chemokine ligand 2,and C-X-C motif chemokine ligand 2),and western blot was performed to detect the expression of peroxiredoxin 1,inducible nitric oxide synthase,and reactive oxygen/mitogen-activated protein kinase signaling pathway proteins.Mouse microglia BV2 were treated in two groups:the control group was stimulated by hydrogen peroxide for 4 hours,and the knockout group was stimulated by hydrogen peroxide for 4 hours at 24 hours after knockdown of peroxiredoxin 1.The level of reactive oxygen species was detected by 2,7-dichlorodihydrofluorescein diacetate probe.RESULTS AND CONCLUSION:(1)Results from transcriptome sequencing,western blot and RT-qPCR confirmed that peroxiredoxin 1 expression levels in mouse spinal cord tissues were significantly higher in the spinal cord injury group than the sham-operated group(P＜0.05).(2)Peroxiredoxin 1 knockdown in microglial cells led to decreased expression of peroxiredoxin 1 mRNA and protein(P＜0.05),increased mRNA expression of interleukin 1β,interleukin 6,inducible nitric oxide synthase,tumor necrosis factor α,C-C motif chemokine ligand 2,and C-X-C motif chemokine ligand 2(P＜0.05),increased protein expression of inducible nitric oxide synthase,P-P38,P-JNK and P-ERK proteins(P＜0.05),and increased level of reactive oxygen species(P＜0.05).To conclude,peroxiredoxin 1 regulates microglial inflammation by targeting the reactive oxygen species/mitogen-activated protein kinase signaling pathway.

BACKGROUND:The inflammatory response of microglia is closely related to neuronal survival,regeneration,and functional recovery after spinal cord injury.Peroxiredoxin 1 is not only involved in the regulation of oxidative stress,but also has an important effect on cell proliferation,apoptosis,and inflammatory response.OBJECTIVE:To investigate the role and mechanism of peroxiredoxin 1 in the inflammatory response of microglia following spinal cord injury.METHODS:(1)Twelve female C57BL/6 mice were randomly divided into sham-operated(n=6)and spinal cord injury(n=6)groups.The sham-operated group was not modeled and acute spinal cord injury models were constructed in the spinal cord injury group using the modified Allen's method.Spinal cord tissue at the injured site was taken at 7 days after modeling and transcriptome sequencing was performed to identify differentially expressed genes.The expression of peroxiredoxin 1 in spinal cord tissues was verified using western blot and RT-qPCR.(2)Mouse microglia BV2 were divided into two groups:the control group was stimulated with lipopolysaccharide for 6 hours,and in the knockout group,lipopolysaccharide stimulation was applied for 6 hours at 24 hours after peroxiredoxin 1 was knocked down in the cells.RT-qPCR was performed to detect mRNA expression of peroxiredoxin 1,inflammatory factors(interleukin 1β,interleukin 6,inducible nitric oxide synthase,tumor necrosis factor α,C-C motif chemokine ligand 2,and C-X-C motif chemokine ligand 2),and western blot was performed to detect the expression of peroxiredoxin 1,inducible nitric oxide synthase,and reactive oxygen/mitogen-activated protein kinase signaling pathway proteins.Mouse microglia BV2 were treated in two groups:the control group was stimulated by hydrogen peroxide for 4 hours,and the knockout group was stimulated by hydrogen peroxide for 4 hours at 24 hours after knockdown of peroxiredoxin 1.The level of reactive oxygen species was detected by 2,7-dichlorodihydrofluorescein diacetate probe.RESULTS AND CONCLUSION:(1)Results from transcriptome sequencing,western blot and RT-qPCR confirmed that peroxiredoxin 1 expression levels in mouse spinal cord tissues were significantly higher in the spinal cord injury group than the sham-operated group(P＜0.05).(2)Peroxiredoxin 1 knockdown in microglial cells led to decreased expression of peroxiredoxin 1 mRNA and protein(P＜0.05),increased mRNA expression of interleukin 1β,interleukin 6,inducible nitric oxide synthase,tumor necrosis factor α,C-C motif chemokine ligand 2,and C-X-C motif chemokine ligand 2(P＜0.05),increased protein expression of inducible nitric oxide synthase,P-P38,P-JNK and P-ERK proteins(P＜0.05),and increased level of reactive oxygen species(P＜0.05).To conclude,peroxiredoxin 1 regulates microglial inflammation by targeting the reactive oxygen species/mitogen-activated protein kinase signaling pathway.

Food-derived bioactive peptides (FBPs), particularly those with ten or fewer amino acid residues and a molecular weight below 1300 Da, have gained increasing attention for their safe, diverse structures and specific biological activities. The development of FBP-based functional foods and potential medications depends on understanding their structure‒activity relationships (SARs), stability, and bioavailability properties. In this review, we provide an in-depth overview of the roles of FBPs in treating various diseases, including Alzheimer's disease, hypertension, type 2 diabetes mellitus, liver diseases, and inflammatory bowel diseases, based on the literature from July 2017 to Mar. 2023. Subsequently, attention is directed toward elucidating the associations between the bioactivities and structural characteristics (e.g., molecular weight and the presence of specific amino acids within sequences and compositions) of FBPs. We also discuss in silico approaches for FBP screening and their limitations. Finally, we summarize recent advancements in formulation techniques to improve the bioavailability of FBPs in the food industry, thereby contributing to healthcare applications.
Humans ; Peptides/therapeutic use* ; Structure-Activity Relationship ; Functional Food ; Diabetes Mellitus, Type 2/drug therapy* ; Biological Availability ; Alzheimer Disease/drug therapy* ; Inflammatory Bowel Diseases/drug therapy* ; Hypertension/drug therapy* ; Liver Diseases/drug therapy* ; Bioactive Peptides, Dietary

Objective: To construct a Family with sequence similarity 50 member A(FAM50A) gene knockout mouse insulinoma pancreatic β-cell line Beta-TC-6 using CRISPR/Cas9 gene editing technology and to prepare polyclonal antibodies specifically recognizing FAM50A. Methods: Two guide RNAs(sgRNAs) targeting the FAM50A gene were designed,and a recombinant plasmid expressing blue fluorescent protein(BFP) was constructed for gene knockout.The successfully constructed plasmid was transfected into Beta-TC-6 cells,and BFP-positive single cells were isolated for clonal expansion.The expanded monoclonal cell lines were genotyped by Sanger sequencing,and FAM50A protein expression was assessed by Western blot.Purified human recombinant FAM50A protein was used to immunize New Zealand rabbits for the preparation of a polyclonal antibody.The specificity of the prepared antibody was then validated using the successfully established FAM50A knockout cell line. Results: A monoclonal cell line with a successful knockout of the FAM50A gene was identified.Sanger sequencing confirmed base deletions at the target site.Western blot analysis showed a complete absence of FAM50A protein expression in this cell line.The prepared polyclonal antibody successfully recognized endogenous murine FAM50A protein in wild-type Beta-TC-6 cells and in hTERT-RPE1 cells overexpressing human FAM50A-GFP fusion protein,while no signal was detected in the FAM50A knockout cells. Conclusion This study successfully established a FAM50A gene knockout Beta-TC-6 cell model and generated a FAM50A polyclonal antibody,providing powerful tools for future research.

Etiological research is necessary for understanding the occurrence and epidemiological patterns of diseases, and is also a prerequisite for the diagnosis, prevention and treatment of clinical diseases. Mendelian randomization(MR), a method of research that combines genetics and epidemiology, has the advantage of exploring the causal relationship between exposure and disease genetically as well as avoiding confounding factors and reverse causation. Thus, it has been extensively utilized in the etiological study of diseases. This paper reviews the implementation of MR in the research of ocular diseases and provides ideas and approaches for the investigation of related mechanisms as well as the development of intervention strategies.

Anti-motion artifact is one of the most important properties of ambulatory ECG monitoring equipment. At present, there is a lack of standardized means to test the performance of anti-motion artifact. ECG simulator and special conductive leather are used to build the simulator, it is used to simulate human skin, to generate ECG signal input for the ECG monitoring equipment attached to it. The mechanical arm and fixed support are used to build a motion simulation system to fix the conductive leather. The mechanical arm is programmed to simulate various motion states of the human body, so that the ECG monitoring equipment can produce corresponding motion artifacts. The collected ECG signals are read wirelessly, observed, analyzed and compared, and the anti-motion artifact performance of ECG monitoring equipment is evaluated. The test results show that by artificially creating the small difference between the two groups of ambulatory ECG monitoring equipment, the system can accurately test the interference signals introduced under the conditions of controlled movement such as tension and torsion, and compare the advantages and disadvantages. The research shows that the test system can provide convenient and accurate verification means for the research of optimizing anti-motion interference.
Humans ; Artifacts ; Signal Processing, Computer-Assisted ; Electrocardiography, Ambulatory/methods* ; Electrocardiography ; Motion

Objective:To evaluate a self-designed intelligent robot-assisted minimally invasive reduction system in the reduction of unstable pelvic fractures by a cadaveric anatomic study.Methods:Ten unembalmed cadavers (7 male and 3 female ones) were used in this study. In each cadaveric specimen an unstable pelvic fracture was created in accordance with clinical case models (3 cases of type B1, 4 cases of type B2 and 3 cases of type C1 by the Tile classification). A self-designed intelligent robot-assisted minimally invasive reduction system was used to assist the reduction in the cadaveric models. Intraoperative registration and navigation time, autonomous reduction time, total operation time and reduction error were measured.Results:Effective reduction was completed in 10 bone models with the assistance of our self-designed intelligent robot-assisted minimally invasive reduction system. The time for intraoperative registration and navigation averaged 47.4 min (from 32 to 74 min), the autonomous reduction time 73.9 min (from 48 to 96 min), and the total operation time 121.3 min (from 83 to 170 min). The reduction error averaged 2.02 mm (from 1.67 to 2.62 mm), and the reduction results met the clinical requirements.Conclusion:Our self-designed intelligent robot-assisted minimally invasive reduction system is a new clinical solution for unstable pelvic fractures, showing advantages of agreement with clinical operative procedures, high reduction accuracy and operational feasibility, and reduced radiation exposure compared to a conventional operation.

Objective To establishing an immortal cell line of familial papillary thyroid carcinoma (FPTC), and explore a new approach for studying familial non-medullary thyroid carcinoma (FNMTC). Methods The specimen from a patient with FPTC was selected, separated, and the primary cells were cultured using DMEM/F12 medium (with TSH, T3, EGF and hydrocortisone). To inducing cell immortalization, the exogenous genes SV40T/TERT were transfected into cells by two ways. RT-PCR was used to detect the expressions of thyroid peroxidase (TPO), thyroid globulin (TG), thyroid stimulating hormone receptor (TSHR) and sodium/iodide co-transporter (NIS). Immunofluorescence method was used to detect the expressions of TPO and GPC3. In order to detect the genomic mutations, the peripheral blood DNA of the patient was extracted. The cell genome was detected. Results The FPTC cells adhered to the plate and showed an irregular polygon shape. The cells can stably grow for six months, FPTC-S (with SV40T transfected) passaged to p26, FPTC cells passaged to p23 and FPTC-ST (with SV40T/TERT transfected) passaged to p19. Both FPTC-S and FPTC-ST can stably express TPO, TG and TSHR in mRNA level. MLH1 R217C mutation existed in the peripheral blood of the patient, and BRAF V600E mutation existed in the primary cultured cells. Either the primary or the immortal cells showed MLH1 R217C mutation. Conclusion This study preliminarily established an immortal cell line of familial papillary thyroid carcinoma with MLH 1 R217C and BRAF V600E mutations. This cell line provides a research model for studying these mutations in FPTC.

Aim To investigate the protective effects of high concentration fentanyl on the brain slice injury induced by oxygen glucose deprivation(OGD).Methods Rat brain slices were made and randomly assigned to four groups:control(n=10),OGD(n=10),fentanyl 50 ?g?L~(-1)(F_(50),n=10) and fentanyl 500 ?g?L~(-1)(F_(500),n=10).Changes of the neuron injury and apoptosis were observed with TTC staining,LDH releases,TUNEL staining,immunohistochemistry and electromicroscope.In addition,changes of intracellular calcium were measured with confocal laser-scanning microscopy.Results F_(50) and F_(500) attenuated the decrease of TTC staining and the increase of LDH release induced by OGD in brain slices.Neuronal apoptosis and changes of neuronal ultrastructures were attenuated by F_(50) and F_(500).Bcl-2 and Bax protein expressions were increased after OGD.Bax protein expression was decreased by F_(50) and F_(500),while Bcl-2 protein expression was increased by F_(50)and F_(500).Intracellular calcium concentration was increased by OGD and then it was lowered by F_(50) and F_(500).The protective effects of F_(50) were more obvious than that of F_(500).Conclusions High concentrations of fentanyl have neuron protective effects against OGD injury in rat brain slices,and fentanyl 50 ?g?L~(-1) has more obvious protective effects than fentanyl 500 ?g?L~(-1).

Objective　 To investigate gene rearrangement and p53 expression i n defining the nature of angioimmunoblastic lymphadenopathy. Methods　 DNA was ext racted from paraffin-embedded tissue blocks of 44 angioimmunoblastic lymphaden o pathy (AIL) patients and analyzed with polymerase chain reaction (PCR) for IgH and TCRγ gene rearrangement. Immunohistochemistry staining was used to detect p53 protein expression. Thirty-five cases were followed-up. Results　 12 out of 44 cases(27.3%) showed TCRγ gene rearr angement and 2 (4.5%) showed IgH gene rearrangement. Rearrangement of both IgH a nd TCRγ genes were detected in 2 cases(4.5%). 14 cases (31.8%) showed p53 posit i ve expression, among which 12 showed positive rearrangement and 2 showed negative (P<0.01). Eight out of 11 patients of p os itive gene rearrangement died in one year, while only 3 patients were still aliv e at the eighteenth month of follow-up, three of 24 patients of negative gene r earrangement were found dead at the time of the one year follow-up, while the r est 21 patients were alive and the longest survival time was 96 months. Conclusions　 Gene rearrangement can define th e pathological nature of AIL. The expression of p53 is highly related to gene r earrangement, and thus an important immunological marker in research on AIL.