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 investigate the frequency and related factors of ineffective triggering (IT) and double triggering (DT) in patients with acute brain injury undergoing invasive mechanical ventilation. METHODS: A retrospective cohort study was conducted using data from a single-center observational trial. Patients with acute brain injury [traumatic brain injury, stroke, and post-craniotomy for brain tumors] undergoing mechanical ventilation in the intensive care unit (ICU) of Beijing Tiantan Hospital, Capital Medical University between June 2017 and July 2019 were retrospectively analyzed. Demographic and clinical data were collected. Respiratory parameters and waveforms during the first 3 days of mechanical ventilation were recorded, with 15-minute waveform segments collected 4 times daily. Airway occlusion pressure (P_0.1) was measured via end-expiratory hold at the end of each recording. IT and DT were identified based on airway pressure, flow, and esophageal pressure waveforms, and the ineffective triggering index (ITI) and DT incidence were calculated. Multivariate Logistic regression was used to identify factors associated with IT and DT. RESULTS: A total of 94 patients with acute brain injury were ultimately enrolled, including 19 cases of traumatic brain injury (20.2%), 39 cases of stroke (41.5%), and 36 cases of post-craniotomy for brain tumor (38.3%). Supratentorial injury was observed in 49 patients (52.1%), while infratentorial injury was identified in 45 patients (47.9%). A total of 94 patients with 1 018 datasets were analyzed; 684 (67.2%) datasets were on pressure support ventilation (PSV), and 334 (32.8%) were on mandatory ventilation. IT was detected in 810 (79.6%) datasets, with a median incidence of 2.1% (0.3%, 12.0%). Datasets demonstrating IT were characterized by lower P_0.1, higher tidal volume (VT), reduced respiratory rate (RR), and decreased minute ventilation (MV) compared to those without IT. The proportion of datasets exhibiting IT was higher during PSV than in mandatory ventilation [83.8% (573/684) vs. 71.0% (237/334), P < 0.05], while, the prevalence of ITI ≥ 10% was lower [23.8% (163/684) vs. 33.5% (112/334), P < 0.05]. DT was detected in 305 datasets (30%), with a median incidence of 0.6% (0.4%, 1.3%). Datasets exhibiting DT were characterized by higher VT, reduced RR, and lower pressure support levels. The incidence of DT was lower in PSV compared to mandatory ventilation modes [0% (0%, 0.3%) vs. 0% (0%, 0.5%), P < 0.05]. The post-craniotomy for brain tumors group exhibited higher ITI, lower RR, reduced MV, and a greater proportion of infratentorial lesions, compared to the TBI group. The infratentorial lesion group demonstrated higher ITI and incidence of DT compared to the supratentorial lesion group [ITI: 3.1% (0.7%, 17.8%) vs. 1.5% (0%, 8.3%), incidence of DT: 0% (0%, 0.5%) vs. 0% (0%, 0%), both P < 0.05]. After adjusting for confounding factors through multivariate logistic regression analysis, infratentorial lesion [odds ratio (OR) = 2.029, 95% confidence interval (95%CI) was 1.465-2.811, P < 0.001], lower P_0.1 (OR = 0.714, 95%CI was 0.616-0.827, P < 0.001), and mandatory ventilation (OR = 1.613, 95%CI was 1.164-2.236, P = 0.004) were independently associated with IT. Additionally, infratentorial lesion (OR = 1.618, 95%CI was 1.213-2.157, P = 0.001), large tidal volume (OR = 1.222, 95%CI was 1.137-1.314, P < 0.001), lower pressure support levels (OR = 0.876, 95%CI was 0.829-0.925, P < 0.001), and mandatory ventilation (OR = 2.750, 95%CI was 1.983-3.814, P < 0.001) were independently associated with DT. CONCLUSION IT and DT were common in patients with acute brain injury. Infratentorial lesions and mandatory ventilation were independently associated with both IT and DT.
Humans ; Respiration, Artificial/methods* ; Retrospective Studies ; Brain Injuries/therapy* ; Intensive Care Units ; Male ; Female ; Middle Aged ; Brain Injuries, Traumatic/therapy* ; Logistic Models ; Aged ; Adult

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:Researches show that a combination of platelet-rich plasma and adipose-derived stem cells can accelerate the healing of skin lesions.However,systematic evidence for the combination of the two is still lacking.The purpose of this study was to assess the efficacy of a combination of two interventions in a clinical rodent skin wound model.METHODS:We searched PubMed,Embase,Cochrane,and CNKI and selected the studies of platelet-rich plasma,adipose-derived stem cell transplantation,or their combination on skin wounds in experimental animals published until July 2023.Wound healing and wound transformation growth factor β,CD31,type Ⅰ collagen,and vascular endothelial growth factor were used as indicators.RevMan 5.3 and Stata 15.0 were used to analyze the data.RESULTS:A total of 12 studies were included,of which 8 studies used rats as experimental subjects and 4 studies used mice as experimental subjects.The experimental group was treated with platelet-rich plasma combined with adipose-stem cell transplantation,and the control group was treated with platelet-rich plasma alone.The results of meta-analysis showed that the wound healing rate of the experimental group at 3,7,and 10 days after treatment was greater than that of the control group[SMD=2.65,95％CI(1.29,4.01),Z=3.81,P=0.0001;SMD=3.38,95％CI(2.47,4.30),Z=7.24,P<0.00001;SMD=2.62,95％CI(1.50,3.73),Z=4.61,P<0.00001].The wound healing time of the experimental group was shorter than that of the control group[SMD=-2.12,95％CI(-3.5,-0.74),P=0.003].The expression of transforming growth factor β,positive rate of CD31,expression of type Ⅰ collagen,and vascular endothelial growth factor in wound of experimental group were higher than those of control group[SMD=5.65,95％CI(1.22,10.08),Z=2.50,P=0.01;SMD=2.49,95％CI(1.96,3.02),Z=9.28,P<0.00001;SMD=3.44,95％CI(0.72,6.17),Z=2.48,P=0.01;SMD=2.38,95％CI(0.97,3.79),Z=3.30,P=0.0010].CONCLUSION:Our results show that platelet-rich plasma+adipose-derived stem cells combined treatment can improve the wound healing rate,shorten the wound healing time,and at the same time increase the expression of transforming growth factor β,CD31,type Ⅰ collagen,and vascular endothelial growth factor to accelerate healing.Due to the limitations of the model,more animal testing and clinical trials are needed.

OBJECTIVE To explore the effect and mechanism of Huiyang Shengji Decoction on wound healing in mice with dia-betic yin syndrome.METHODS SPF C57BL/6 mice were selected,and 24 diabetic ulcer models were established by intraperitoneal injection of streptozotocin and skin defect method.The mice were randomly divided into model group,low-dose,medium-dose and high-dose Huiyang Shengji Decoction groups,with 6 mice in each group.Six mice were selected to establish a common wound model as the blank group.The blank group and model group were treated with distilled water by intragastric administration every day,and the Huiyang Shengji Decoction groups were treated with Huiyang Shengji Decoction by intragastric administration every day.The wounds were photographed every day,and the wound healing rate was recorded.HE staining was used to observe the growth of granulation tis-sue in the wounds.Western blot was used to detect the expression of Axl,Tyro3 and Mertk proteins related to wound efferocytosis.Im-munohistochemistry was used to detect the expression of CD11c in wound tissue.qPCR was used to detect the mRNA expression of TNF-α,IL-1β,IL-10 and TGF-β1 in the wounds.TUNEL staining was used to calculate the number of apoptotic cells(AC)in the wounds.Mouse primary bone marrow dendritic cells(BMDC)were resuspended in 1640 medium containing 10%blank serum or 5%,10%,and 20%Huiyang Shengji Decoction-containing serum.After culturing in a cell culture incubator at 37℃and 5%CO2 for 24 h,the wound endocytosis-related receptors were detected by Western blot.BMDC and AC were co-cultured in vitro,and the phag-ocytic rate of BMDC phagocytosis of AC was detected by flow cytometry.RESULTS The wound healing in each dose group of Huiyang Shengji Decoction was significantly faster than that in the model group(P<0.05).Compared with the model group,the protein expres-sion of Axl and Tyro3 in the wound tissue of mice in the high dose group of Huiyang Shengji Decoction was significantly increased(P<0.01,P<0.000 1),and there was no significant difference in the expression of Mertk protein in the wound tissue of mice in the low,medium and high dose groups(P>0.05).The Huiyang Shengji Decoction significantly inhibited the expression of IL-1β and TNF-α mRNA in the wound tissue(P<0.05),promoted the expression of TGF-β1 and IL-10 mRNA(P<0.05),and the number of AC in the wound after treatment with Huiyang Shengji Decoction was less than that in the model group(P<0.05).In the in vitro experiment,compared with the model group,the protein expression of Axl and Mertk in BMDC in the 20%Huiyang Shengji Decoction-containing serum group was significantly increased(P<0.01,P<0.001),and the protein expression of Tyro3 in the 5%,10%,and 20%Huiy-ang Shengji Decoction-containing serum groups was higher than that in the model group(P<0.01,P<0.001).The phagocytic rate of AC in the Huiyang Shengji Decoction group was higher than that in the model group(P<0.05).CONCLUSION Huiyang Shengji Decoction can enhance the efferocytosis function of dendritic cells(DC),reduce AC aggregation in the wound,promote the disappear-ance of wound inflammation and tissue repair,and accelerate the healing of skin wounds.