Background: In acute and chronic renal inflammatory diseases, the activation of inflammatory cells is involved in the defect of erythropoietin (EPO) production. Ras guanine nucleotide-releasing protein-4 (RasGRP4) promotes renal inflammatory injury in type 2 diabetes mellitus (T2DM). Our study aimed to investigate the role and mechanism of RasGRP4 in the production of renal EPO in diabetes. Methods: The degree of tissue injury was observed by pathological staining. Inflammatory cell infiltration was analyzed by immunohistochemical staining. Serum EPO levels were detected by enzyme-linked immunosorbent assay, and EPO production and renal interstitial fibrosis were analyzed by immunofluorescence. Quantitative real-time polymerase chain reaction and Western blotting were used to detect the expression of key inflammatory factors and the activation of signaling pathways. In vitro, the interaction between peripheral blood mononuclear cells (PBMCs) and C3H10T1/2 cells was investigated via cell coculture experiments. Results: RasGRP4 decreased the expression of hypoxia-inducible factor 2-alpha (HIF2A) via the ubiquitination–proteasome degradation pathway and promoted myofibroblastic transformation by activating critical inflammatory pathways, consequently reducing the production of EPO in T2DM mice. Conclusion RasGRP4 participates in the production of renal EPO in diabetic mice by affecting the secretion of proinflammatory cytokines in PBMCs, degrading HIF2A, and promoting the myofibroblastic transformation of C3H10T1/2 cells.

Background: In acute and chronic renal inflammatory diseases, the activation of inflammatory cells is involved in the defect of erythropoietin (EPO) production. Ras guanine nucleotide-releasing protein-4 (RasGRP4) promotes renal inflammatory injury in type 2 diabetes mellitus (T2DM). Our study aimed to investigate the role and mechanism of RasGRP4 in the production of renal EPO in diabetes. Methods: The degree of tissue injury was observed by pathological staining. Inflammatory cell infiltration was analyzed by immunohistochemical staining. Serum EPO levels were detected by enzyme-linked immunosorbent assay, and EPO production and renal interstitial fibrosis were analyzed by immunofluorescence. Quantitative real-time polymerase chain reaction and Western blotting were used to detect the expression of key inflammatory factors and the activation of signaling pathways. In vitro, the interaction between peripheral blood mononuclear cells (PBMCs) and C3H10T1/2 cells was investigated via cell coculture experiments. Results: RasGRP4 decreased the expression of hypoxia-inducible factor 2-alpha (HIF2A) via the ubiquitination–proteasome degradation pathway and promoted myofibroblastic transformation by activating critical inflammatory pathways, consequently reducing the production of EPO in T2DM mice. Conclusion RasGRP4 participates in the production of renal EPO in diabetic mice by affecting the secretion of proinflammatory cytokines in PBMCs, degrading HIF2A, and promoting the myofibroblastic transformation of C3H10T1/2 cells.

Background: In acute and chronic renal inflammatory diseases, the activation of inflammatory cells is involved in the defect of erythropoietin (EPO) production. Ras guanine nucleotide-releasing protein-4 (RasGRP4) promotes renal inflammatory injury in type 2 diabetes mellitus (T2DM). Our study aimed to investigate the role and mechanism of RasGRP4 in the production of renal EPO in diabetes. Methods: The degree of tissue injury was observed by pathological staining. Inflammatory cell infiltration was analyzed by immunohistochemical staining. Serum EPO levels were detected by enzyme-linked immunosorbent assay, and EPO production and renal interstitial fibrosis were analyzed by immunofluorescence. Quantitative real-time polymerase chain reaction and Western blotting were used to detect the expression of key inflammatory factors and the activation of signaling pathways. In vitro, the interaction between peripheral blood mononuclear cells (PBMCs) and C3H10T1/2 cells was investigated via cell coculture experiments. Results: RasGRP4 decreased the expression of hypoxia-inducible factor 2-alpha (HIF2A) via the ubiquitination–proteasome degradation pathway and promoted myofibroblastic transformation by activating critical inflammatory pathways, consequently reducing the production of EPO in T2DM mice. Conclusion RasGRP4 participates in the production of renal EPO in diabetic mice by affecting the secretion of proinflammatory cytokines in PBMCs, degrading HIF2A, and promoting the myofibroblastic transformation of C3H10T1/2 cells.

Background: In acute and chronic renal inflammatory diseases, the activation of inflammatory cells is involved in the defect of erythropoietin (EPO) production. Ras guanine nucleotide-releasing protein-4 (RasGRP4) promotes renal inflammatory injury in type 2 diabetes mellitus (T2DM). Our study aimed to investigate the role and mechanism of RasGRP4 in the production of renal EPO in diabetes. Methods: The degree of tissue injury was observed by pathological staining. Inflammatory cell infiltration was analyzed by immunohistochemical staining. Serum EPO levels were detected by enzyme-linked immunosorbent assay, and EPO production and renal interstitial fibrosis were analyzed by immunofluorescence. Quantitative real-time polymerase chain reaction and Western blotting were used to detect the expression of key inflammatory factors and the activation of signaling pathways. In vitro, the interaction between peripheral blood mononuclear cells (PBMCs) and C3H10T1/2 cells was investigated via cell coculture experiments. Results: RasGRP4 decreased the expression of hypoxia-inducible factor 2-alpha (HIF2A) via the ubiquitination–proteasome degradation pathway and promoted myofibroblastic transformation by activating critical inflammatory pathways, consequently reducing the production of EPO in T2DM mice. Conclusion RasGRP4 participates in the production of renal EPO in diabetic mice by affecting the secretion of proinflammatory cytokines in PBMCs, degrading HIF2A, and promoting the myofibroblastic transformation of C3H10T1/2 cells.

Objective: To investigate the precise detection methods for weak partial D type 15 and evaluate their implications for blood transfusion safety, along with the development of corresponding strategies. Methods: A combination of serological methods, including the microplate method, indirect antiglobulin tube method, and microcolumn gel card method, was employed to identify RhD-negative and RhD variant samples. RhD-negative samples were screened for the presence of RHD genes using whole-blood direct PCR amplification. Subsequently, RhD variant samples and RhD-negative samples containing RHD genes underwent full-coding-region sequencing of the RHD gene to confirm their genotypes. The genotyping results were further correlated with the serological test findings for comprehensive analysis. Results: Among 615 549 first-time healthy blood donors, 3 401 samples with an RhD-negative phenotype and 156 samples with RhD variant were identified. Of the 3 401 RhD-negative samples, 1 054 were found to harbor RHD genes. Gene sequencing analysis of the 156 RhD variants and the 1 054 serological negative samples revealed that 89 samples contained the RHD 15 (c. 845G>A) allele. Conclusion: The integration of serological testing methods and genotyping technologies for the precise determination of RhD blood type plays a critical role in ensuring the safety and compatibility of blood transfusions.

Objective To study the impact of a Western diet-type feed on biological indicators and histopathology in APOE-/-mice.Methods Forty-eight female and 48 male APOE--mice,and 48 female and 48 male C57BL/6J mice were divided into eight breeding groups:APOE-/-breeding feed group,APOE-/-Western dietary feed group,C57BL/6J breeding feed group,and C57BL/6J Western dietary feed group(24 male and 24 female mice per group).Mice were fed the respective diets from 3 weeks until the end of the experiment at 20 weeks.After the experiment,serum was collected for measurement of biochemical indicators.Aortas were removed for oil red O staining and gross examination and the aorta root was paraffin sectioned and stained with hematoxylin and eosin.Results A Western diet did not significantly increase body weight in APOE-/-mice,but did significantly improve the blood lipid index and total cholesterol,low-density lipoprotein,and high-density lipoprotein levels,and promoted the formation of atherosclerotic plaques.Male mice were suitable for modeling gross aortic plaques while female mice were suitable for modeling aortic arch root plaques.Conclusions A Western diet can promote atherosclerosis in APOE-/-mice,increase the aortic plaque area ratio,shorten modeling time,and improve modeling uniformity.

Objective:To investigate the risk factors for aneurysm rupture and post-intervention complications in intracranial arterial stenosis patients with intracranial aneurysms.Methods:A retrospective analysis was performed; 238 intracranial arterial stenosis patients with intracranial aneurysms (306 intracranial aneurysms) admitted to Cerebrovascular Disease Department, Neurosurgery Center, Zhujiang Hospital, Southern Medical University from January 2018 to August 2022 were chosen. Ruptured group and unruptured group were divided according to the rupture of intracranial aneurysms. Additionally, 139 patients who underwent interventional therapy and had complete follow-up data were divided into 2 groups according to occurrence of post-intervention complications. Univariate and multivariate Logistic regression analyses were used to identify the risk factors for aneurysm rupture and post-intervention complications.Results:(1) Of 238 patients, 269 unruptured aneurysms and 37 ruptured aneurysms were noted. Univariate regression analysis showed that significant difference was noted between the ruptured group and unruptured group in female ratio, aneurysm distribution, proportion of irregular shaped aneurysms, percentages of patients with increased white blood cell count, neutrophil count, total cholesterol and D-2 polymer, and percentage of patients with decreased blood lymphocyte count ( P<0.05). Multivariate Logistic regression analysis showed that irregular shaped aneurysms ( OR=12.393, 95% CI: 4.114-37.332, P<0.001), elevated neutrophil count ( OR=18.753, 95% CI: 6.555-53.648, P<0.001), and increased D-2 polymer ( OR=4.410, 95% CI: 1.758-11.065, P=0.002) were independent risk factors for aneurysm rupture in intracranial arterial stenosis patients with intracranial aneurysms. (2) Of the 139 patients, 57 had complications and 82 had no complications. Univariate regression analysis showed that the proportion of patients with hypertension history, distribution of arterial stenosis, and proportion of patients with elevated blood D-2 polymer were significantly different between patients with and without complications ( P<0.05); while multivariate Logistic regression analysis did not identify these 3 indexes as independent risk factors for post-intervention complications ( P>0.05). Conclusion:Patients with irregular shaped aneurysms, elevated blood neutrophil count and D-2 polymer trend to have aneurysm rupture; hypertension history, arterial stenosis, and elevated D-2 polymer have impact on postoperative complications in intracranial arterial stenosis patients with intracranial aneurysms.

Objective To explore the effects of different types of drinking water on the growth and fecal flora of mice.Methods Specific pathogen-free NIH mice were randomly divided into five groups,32 mice each group,with half males and half females in each group.The group were given either purified water(control group),acidified water,alkalized water,weakly acidic water or solid water.Diet and body weight were monitored continuously for 20 days.After the experiment,animal fecal samples were collected,and the V3-V4 region was amplified with bacterial 16S rDNA universal primers.An Illumina Miseq high-throughput sequencing platform was used for high-throughput sequencing,and microbial community,α diversity and β diversity were analyzed by bioinformatics method.Results The body weight of female mice given different pH values of weakly acidic water was higher,while the weight of the other groups was lower,than that of the control group(P＞0.05).The body weight of male mice in the acidified water group was higher,while that of other groups was lower,than that of control group,but there was no statistical difference between the groups(P＞0.05).The body weights of male and female mice in the solid water group were lower than those in the control group(P＜0.05).The food and water intake of the female animals in the alkaline water group and the water intake of female animals in the solid water group were lower than those of the other groups.OTU clustering analysis showed that the data volume of the sequencing was reasonable,and the fecal flora species of NIH mice were divided into five phyla,among which Bacteroides and Firmicutes were dominant.Unclassified Pseudopurpuromonas,Lactobacillus and Alistipes were the main genera.There were differences in fecal flora abundance and diversity among the mice given the five drinking water types.α analysis showed that the acidified water group had the highest flora abundance and diversity,while the solid water group had the lowest flora diversity.β analysis showed that the fecal flora composition in the solid water group was the closest to that of the control group,followed by the alkalized water group,acidified water group and weakly acidic water group.Conclusions Through an exploration of the effects of consuming different forms of water,this study revealed that solid water consumption had the greatest effects on body weight,feed intake,water consumption,and fecal flora of mice.The abundance and diversity of fecal flora in mice were affected by different pH values of drinking water,especially acidified water.

Objective To investigate the serological characteristics and molecular mechanism of an individual with Am phenotype.Methods The sample with ABO blood group discrepancy was confirmed by serological techniques.The full cod-ing and flanking regions of the ABO gene including intron 1 transcription factor binding site were identified through direct se-quencing of PCR-amplified products.PCR products of exon 6-7 were validated to isolate the ABO gene haplotypes by clo-ning and sequencing individual colonies.Bioinformatics software was used to analyze the structure of the mutant protein.Re-sults The serologic characteristics of ABO blood typing showed the rare Am phenotype.The c.467C/T and c.912C/A heter-ozygous sites in exon 7 were identified by direct sequencing analysis.Further TA cloning and sequencing revealed that the patient carried an ABO*O.01.01 allele and a novel ABO*A allele.The new allele sequence had one nucleotide alteration(C＞A)at position 912 on the background of the ABO*A1.02 allele.The new allele sequence has been included in the Gen-Bank database with the entry number JX489776.The c.912C＞A mutation was predicted to be"probably damaging"and"deleterious"by PolyPhen2 and PROVEAN algorithms,respectively.The free energy change(ΔΔG)value predicted it to have a destabilizing effect on the GTA protein.Meanwhile,modeling of the 3D structure predicted that the p.S304R amino acid substitution may alter the hydrogen bond of the GTA protein.Conclusion The p.S304R substitution of α-1,3-N-acetylgalactosaminyltransferase gene may reduce the antigen expression owing to a greatly destabilizing effect on the structure and function of the GTA protein.

With the continuous advancement of modern research on tongue diagnosis in traditional Chinese medicine,tongue diagnostic equipment plays an undeniable role in promoting the objectification of tongue diagnosis.Traditional Chinese medicine tongue diagnosis is only the subjective judgment of clinical doctors on tongue color,coating,etc.,while digital tongue diagnostic equipment and digital image processing technology can visually display tongue color,coating,etc.through digital representation,obtain the color distribution of the tongue body,saturation,brightness,and other parameter values of tongue color,objectively reflecting physiological and pathological changes in the body.The article briefly introduces the main working principles,technical parameters,image segmentation processing methods,and tongue image analysis techniques of digital tongue diagnostic equipment.It focuses on summarizing the research status of digital tongue diagnostic equipment in promoting the objectification of traditional Chinese medicine tongue diagnosis in some common diseases,and looks forward to the future application and development of digital tongue diagnostic equipment.