Objective: To investigate the current status of iron metabolism among apheresis donors in Guangzhou and analyze the improvement effects of health education on iron deficiency in frequent apheresis donors. Methods: Using a generalized linear mixed model (GLMM), a 180-day follow-up was conducted on 261 eligible apheresis donors at the Guangzhou Blood Center from January to July 2024. Hemoglobin (Hb), serum ferritin (SF), unsaturated iron-binding capacity (UIBC), total iron-binding capacity (TIBC), and transferrin saturation (TS) were selected as outcome variables. The effects of gender, age group, and number of donations within 180 days on these outcomes were analyzed and modeled. A general linear model (GLM) with repeated measures was applied to 55 donors who received health education interventions, comparing changes in Hb and iron metabolism-related indicators before and after follow-up and health education. Results: No significant difference in Hb levels was observed between first-time and regular apheresis donors, but SF levels were significantly higher in first-time donors (F=6.195, P<0.05). The GLMM revealed that female donors exhibited more significant reductions in Hb (T=-12.546) and SF (T=-5.829)(P<0.05 for both). Age group showed no interactive effects on Hb or SF changes. While number of donations within 180 days had no interactive effect on Hb, SF levels significantly decreased with increased number of donations (using ≥9 donations as the reference group; P<0.05 for all groups). After health education, Hb levels remained unchanged, but SF increased compared to pre-intervention levels (mean difference: -18.571, P<0.05), though a declining trend persisted compared to baseline (mean difference from baseline to post-intervention: 23.068,P<0.05). Conclusion: Female and number of donations are primary factors contributing to SF reduction in apheresis donors. Health education interventions promote SF recovery. Extending donation intervals and reinforcing iron deficiency-related health education may improve iron status in donors.

BACKGROUND: Regulatory dendritic cell (DCreg) subset exhibits a unique capacity for inducing immune tolerance among the variety subsets of dendritic cells (DCs) within gut-associated lymphoid tissues (GALTs). Fms-like tyrosine kinase 3 ligand (FLT3L) is involved in the differentiation of DCregs and the subsequent expansion of regulatory T-cells (Tregs) mediated by DCregs, though the precise mechanism remains poorly understood. This study aimed to explore the expansion mechanism of Treg induced by DCreg and the role of FLT3L in this process. METHODS: DCregs were distinguished from other DC subsets isolated from GALTs of BALB/c mice through a mixed lymphocyte reaction assay. The functions and mechanisms by which FLT3L promoted Treg expansion via DCregs were investigated in vitro through co-culture experiments involving DCregs and either CD4 + CD25 - T-cells or CD4 + CD25 + T-cells. Additionally, an in vivo experiment was conducted using a dextran sulfate sodium (DSS)-induced colitis model in mice. RESULTS: CD103 + CD11b + DC exhibited DCreg-like functionality and was identified as DCreg for subsequent investigation. Analysis of Foxp3 + Treg percentages within a co-culture system of CD4 + CD25 - T-cells and DCregs, with or without FLT3L, demonstrated the involvement of the FLT3/FLT3L axis in driving the differentiation of precursor T-cells into Foxp3 + Tregs induced by DCregs. Cell migration and co-culture assays revealed that the FLT3/FLT3L axis enhanced DCreg migration toward Tregs via the Rho pathway. Additionally, it was observed that DCregs could promote Treg proliferation through the Notch pathway, as inhibition of Notch signaling by DAPT (N-[N-(3,5-difluorophenacetyl)-l-alanyl]-S-phenylglycine t-butyl ester) suppressed Treg expansion within the co-culture system of DCregs and CD4 + T-cells or CD4 + CD25 + T-cells. Furthermore, the FLT3/FLT3L axis influenced JAG1 expression in DCregs, indirectly modulating Treg expansion. In vivo experiments further established that FLT3L promoted DCreg expansion and restored Treg balance in DSS-induced colitis models, thereby ameliorating colitis symptoms in mice. CONCLUSION The FLT3/FLT3L axis is integral to the maintenance of DCreg function in Treg expansion.
Animals ; T-Lymphocytes, Regulatory/immunology* ; Dendritic Cells/immunology* ; Mice ; Mice, Inbred BALB C ; Membrane Proteins/metabolism* ; Receptors, Notch/metabolism* ; Lymphoid Tissue/metabolism* ; Signal Transduction/physiology* ; Coculture Techniques ; Flow Cytometry

OBJECTIVE: To investigate the Wnt signaling pathway and miRNAs mechanism of extracts of Plastrum Testudinis (PT) in the treatment of osteoporosis (OP). METHODS: Thirty female Sprague Dawley rats were randomly divided into 5 groups by random number table method, including sham group, ovariectomized group (OVX), ovariectomized groups treated with high-, medium-, and low-dose PT (160, 80, 40 mg/kg per day, respectively), with 6 rats in each group. Except for the sham group, the other rats underwent bilateral ovariectomy to simulate OP and received PT by oral gavage for 10 consecutive weeks. After treatment, bone mineral density was measured by dual-energy X-ray absorptiometry; bone microstructure was analyzed by micro-computed tomography and hematoxylin and eosin staining; and the expressions of osteogenic differentiation-related factors were detected by immunochemistry, Western blot, and quantitative polymerase chain reaction. In addition, Dickkopf-1 (Dkk-1) was used to inhibit the Wnt signaling pathway in bone marrow mesenchymal stem cells (BMSCs) and miRNA overexpression was used to evaluate the effect of miR-214 on the osteogenic differentiation of BMSCs. Subsequently, PT extract was used to rescue the effects of Dkk-1 and miR-214, and its impacts on the osteogenic differentiation-related factors of BMSCs were evaluated. RESULTS: PT-M and PT-L significantly reduced the weight gain in OVX rats (P<0.05). PT also regulated the bone mass and bone microarchitecture of the femur in OVX rats, and increased the expressions of bone formation-related factors including alkaline phosphatase, bone morphogenetic protein type 2, collagen type I alpha 1, and runt-related transcription factor 2 when compared with the OVX group (P<0.05 or P<0.01). Meanwhile, different doses of PT significantly rescued the inhibition of Wnt signaling pathway-related factors in OVX rats, and increased the mRNA or protein expressions of Wnt3a, β-catenin, glycogen synthase kinase-3β, and low-density lipoprotein receptor-related protein 5 (P<0.05 or P<0.01). PT stimulated the osteogenic differentiation of BMSCs inhibited by Dkk-1 and activated the Wnt signaling pathway. In addition, the expression of miR-214 was decreased in OVX rats (P<0.01), and it was negatively correlated with the osteogenic differentiation of BMSCs (P<0.01). MiR-214 mimic inhibited Wnt signaling pathway in BMSCs (P<0.05 or P<0.01). Conversely, PT effectively counteracted the effect of miR-214 mimic, thereby activating the Wnt signaling pathway and stimulating osteogenic differentiation in BMSCs (P<0.05 or P<0.01). CONCLUSION PT stimulates bone formation in OVX rats through β-catenin-mediated Wnt signaling pathway, which may be related to inhibiting miR-214 in BMSCs.
Animals ; MicroRNAs/genetics* ; Female ; Rats, Sprague-Dawley ; Wnt Signaling Pathway/genetics* ; Osteogenesis/genetics* ; Mesenchymal Stem Cells/cytology* ; Cell Differentiation/drug effects* ; Bone Density/drug effects* ; Ovariectomy ; Osteoporosis/drug therapy* ; beta Catenin/metabolism* ; Rats ; Intercellular Signaling Peptides and Proteins/metabolism* ; Drugs, Chinese Herbal/pharmacology*

OBJECTIVE: To investigate the predictive value of oxygenation index (PaO₂/FiO₂) at intensive care unit (ICU) admission on 30-day mortality in patients with sepsis. METHODS: A retrospective study was conducted. Patients with sepsis who were hospitalized in the ICU of the Affiliated Hospital of Jining Medical University from April 2015 to October 2023 were enrolled. The demographic information, comorbidities, sites of infection, vital signs and laboratory test indicators at the time of admission to the ICU, disease severity scores within 24 hours of admission to the ICU, treatment process and prognostic indicators were collected. According to the PaO₂/FiO₂ at ICU admission, patients were divided into Q1 group (PaO₂/FiO₂ of 4.1-16.4 cmHg, 1 cmHg ≈ 1.33 kPa), Q2 group (PaO₂/FiO₂ of 16.5-22.6 cmHg), Q3 group (PaO₂/FiO₂ of 22.7-32.9 cmHg), and Q4 group (PaO₂/FiO₂ of 33.0-94.8 cmHg). Differences in the indicators across the four groups were compared. Multifactorial Cox regression analysis was used to assess the relationship between PaO₂/FiO₂ and 30-day mortality of patients with sepsis. The predictive value of PaO₂/FiO₂, sequential organ failure assessment (SOFA) and acute physiology and chronic health evaluation II (APACHE II) on 30-day prognosis of patients with sepsis was analyzed by receiver operator characteristic curve (ROC curve). RESULTS: A total of 1 711 patients with sepsis were enrolled, including 428 patients in Q1 group, 424 patients in Q2 group, 425 patients in Q3 group, and 434 patients in Q4 group. 622 patients died at 30-day, the overall 30-day mortality was 36.35%. There were statistically significant differences in age, body mass index (BMI), history of smoking, history of alcohol consumption, admission heart rate, respiratory rate, APACHE II score, SOFA score, Glasgow coma score (GCS), site of infection, Combined chronic obstructive pulmonary disease (COPD), blood lactic acid (Lac), prothrombin time (PT), albumin (Alb), total bilirubin (TBil), pH, proportion of mechanical ventilation, duration of mechanical ventilation, proportion of vasoactive medication used, and maximal concentration, length of ICU stay, hospital stay, incidence of acute kidney injury, in-hospital mortality, 30-day mortality among the four groups. Multivariate Cox regression analysis showed that after adjusting for confounding factors, for every 1 cmHg increase in PaO₂/FiO₂ at ICU admission, the 30-day mortality risk decreased by 2% [hazard ratio (HR) = 0.98, 95% confidence interval (95%CI) was 0.98-0.99, P < 0.001]. The 30-day mortality risk in the Q4 group was reduced compared with the Q1 group by 41% (HR = 0.59, 95%CI was 0.46-0.76, P < 0.001). The fitted curve showed that a curvilinear relationship between PaO₂/FiO₂ and 30-day mortality after adjustment for confounders. In the inflection point analysis, for every 1 cmHg increase in PaO₂/FiO₂ at PaO₂/FiO₂ < 28.55 cmHg, the risk of 30-day death in sepsis patients was reduced by 5% (HR = 0.95, 95%CI was 0.94-0.97, P < 0.001); when PaO₂/FiO₂ ≥ 28.55 cmHg, there was no statistically significant association between PaO2/FiO2 and the increase in the risk of 30-day death in sepsis (HR = 1.01, 95%CI was 0.99-1.02, P = 0.512). ROC curve analysis showed that the area under the curve (AUC) for the prediction of 30-day mortality by admission PaO₂/FiO₂ in ICU sepsis patients was 0.650, which was lower than the predictive ability of the SOFA score (AUC = 0.698) and APACHE II score (AUC = 0.723). CONCLUSION In patients with sepsis, PaO₂/FiO₂ at ICU admission is strongly associated with 30-day mortality risk, alerting healthcare professionals to pay attention to patients with low PaO₂/FiO₂ for timely interventions.
Humans ; Sepsis/mortality* ; Intensive Care Units ; Retrospective Studies ; Prognosis ; Hospital Mortality ; Oxygen ; Male ; Predictive Value of Tests ; Female ; Middle Aged ; Aged

Background: The function of CD69 expressed on T cells in chronic hepatitis B (CHB) remains unclear. We aimed to elucidate the roles of CD69 on T cells in the disease process and in antiviral therapy for CHB. Methods: We enrolled 335 treatment-naive patients with CHB and 93 patients with CHB on antiviral therapy. CD69, antiviral cytokine production by T cells, T-helper (Th) cells, and inhibitory molecules of T cells were measured using flow cytometry, and clinical-virological characteristics were examined dynamically during antiviral therapy. Results: CD69 expression on CD3+, CD4+, and CD8+ T cells was the lowest in the immune-active phase and was negatively correlated with liver transaminase activity, fibrosis features, inflammatory cytokine production by T cells, and Th-cell frequencies but positively with inhibitory molecules on T cells. CD69 expression on CD3+, CD4+, and CD8+ T cells decreased after 48 weeks of antiviral therapy, and patients with hepatitis B e antigen (HBeAg) seroconversion in week 48 showed lower CD69 expression on T cells at baseline and week 48. The area under the ROC curve of CD69 expression on T cells at baseline for predicting HBeAg seroconversion in week 48 was 0.870, the sensitivity was 0.909, and the specificity was 0.714 (P = 0.002). Conclusions CD69 negatively regulates T-cell immunity during CHB, and its expression decreases with antiviral therapy. CD69 expression predicts HBeAg seroconversion in week 48. CD69 may play an important negative role in regulating T cells and affect the efficacy of antiviral therapy.

Objective To explore the possible therapeutic mechanism of Gastrodin Injection combined with ganglioside in the adjuvant treatment of spinal cord injury(SCI)based on NOD-like receptor thermal protein domain associated protein 3(NLRP3)signaling pathway.Methods A total of 108 patients with SCI were randomly divided into an observation group and a control group,with 54 cases in each group.Both groups were given routine rehabilitation training.Additionally,the control group was treated with intravenous drip of ganglioside,and the observation group was treated with intravenous drip of Gastrodin Injection combined with ganglioside.A course of treatment covered 30 days and the two groups were treated for 2 continuous courses.The time for the recovery of muscle strength,time to leave a sickbed for walking and time to stay in hospital for observation were compared between the two groups.The American Spinal Injury Association(ASIA)score and the serum levels of brain-derived neurotrophic factor(BDNF),central nervous specific protein(S-100β),interleukin(IL)-1β,IL-18,NLRP3,cystein-containing aspartate specific protease 1(Caspase-1)and apoptosis-associated speck-like protein containing a CARD(ASC)in the two groups were observed before and after treatment.After treatment,the clinical efficacy and safety of the two groups were evaluated.Results(1)After two courses of treatment,the total effective rate of the observation group was 83.33%(45/54),and that of the control group was 66.67%(36/54).The intergroup comparison(tested by chi-square test)showed that the curative effect of the observation group was significantly superior to that of the control group(P＜0.05).(2)After treatment,the time for the recovery of muscle strength,time to leave a sickbed for walking and time to stay in hospital for observation in the observation group were significantly shorter than those in the control group(P＜0.01).(3)After one and 2 courses of treatment,the ASIA scores of pain sense,motor sense and tactile sensation in the two groups were significantly higher than those before treatment(P＜0.05),and the scores after 2 courses of treatment were higher than those after one course of treatment(P＜0.05).The intergroup comparison showed that the increase of ASIA scores of pain sense,motor sense and tactile sensation in the observation group after one and 2 courses of treatment was significantly superior to that in the control group(P＜0.05).(4)After one and 2 courses of treatment,the serum BDNF level in the two groups was higher than that before treatment(P＜0.05)and the levels of NLRP3,ACS,Caspase-1 protein and serum IL-1β,IL-18 and S-100β were lower than those before treatment(P＜0.05),and the changes in the levels of above indicators in the two groups after 2 courses of treatment were more obvious than those after one course of treatment(P＜0.05).The intergroup comparison showed that the increase of serum BDNF level and the decrease of the levels of protein NLRP3,ACS,Caspase-1 and serum IL-1β,IL-18 and S-100β in the observation group were significantly superior to those in the control group(P＜0.05).(5)During the treatment,no drug-related adverse reactions occurred in the two groups.Conclusion Gastrodin Injection combined with ganglioside exerts certain effect for the treatment of SCI patients.The combined therapy is effective on relieving the neuroinflammatory response and promoting the recovery of neurological function by regulating the NLRP3 inflammasomes,and its curative effect is superior to that of ganglioside alone.

Objective:To investigate the clinical efficacy of endovascular interventional therapy for spontaneous isolated superior mesenteric artery dissection(SISMAD).Methods:The retrospective cohort study was conducted. The clinical data of 87 patients with SISMAD who were admitted to The First Affiliated Hospital of Army Medical University from March 2012 to March 2023 were collected. There were 80 males and 7 femals, aged 54(49,61)years. Of 87 patients, 55 cases undergoing conservative therapy were allocated into conservative therapy group and 32 cases under-going endovascular interventional therapy were allocated into endovascular interventional therapy group. Observation indicators: (1) clinical characteristics; (2) treatment; (3) follow-up. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the t test. Measurement data with skewed distribution were represented as M( Q1, Q3), and comparison between groups was conducted using the rank sum test. Count data were represented as absolute numbers and comparison between groups was conducted using the chi-square test or Fisher exact probability. Results:(1) clinical characteristics. There were significant differences in the cases with symptoms, percentage of neutrophils between the conservative therapy group and the endovascular interventional therapy group ( P<0.05). There was no significant difference in the proportion of Yun classification between the two groups ( P>0.05). (2)Treatment. There were significant differences in the complete vascular remodeling, duration of hospital stay, and total expenses between the conservative therapy group and the endovascular interventional therapy group ( χ2=23.752, t=-4.213, -16.421, P<0.05). There were 34 patients in the conservative therapy group and 24 patients in the endovascular interventional therapy group with relieved abdominal pain, respectively, showing no significant difference between the two groups ( P>0.05). For symptomatic patients in the conservative therapy group, symptoms including abdominal pain, nausea, vomiting, diarrhea, hematochezia were relieved or disappeared, and no intestinal ischemia or rupture occurred. For patients in the endovascular interventional therapy group, 30 cases were implanted stents, the operation time was 115(86,155)minutes, volume of intraoperative blood loss was 5(5,10)mL, dose of contrast media was (200±51)mL. There were 23, 8 and 1 cases with the contrast medium as Iodoxanol, Ioprosamide, Iodohexanol, respectively. About the surgical methods, 14 patients received single bare stent implantation, 3 cases received bare stent-assisted coil embolization, 10 cases received multiple bare stent implantation, 3 cases received covered stent implantation, 2 cases received angiography alone. A total of 39 self-expandable bare metal stents and 3 self-expandable covered stents were implanted. The diameter and length of the stents were (6.5±1.0)mm and (69±23)mm, respectively. Two asymptomatic patients had failure in endovascular interventional therapy and underwent superior mesenteric artery angiography. For the endovascular interventional therapy group, 92.3%(24/26) of patients were relieved abdominal pain and 2 patients with abdominal pain were improved after symptomatic treatment. (3) Follow-up. All the 87 patients were followed up for 12(4,24)months, without recurrent abdominal pain or secondary intervention. During the follow-up, 82 patients underwent computed tomography angiography or ultrasonography, and 5 patients had no available results. There was no SISMAD related death or superior mesenteric artery rupture. Eight patients in the conservative therapy group achieved complete vascular remodeling, versus 21 cases in the endovascular interventional therapy group, showing a significant difference between the two groups ( χ2=23.752, P<0.05). Conclusions:Compared with conservative therapy, patients undergoing endovascular interventional therapy for SISMAD has loner hospital stay, higher total costs, higher complete vascular remodeling rate. There is no recurrent abdominal pain in two methods.

Gorham-Stout disease(GSD)is a sporadic chronic disease characterized by progressive bone dissolution,absorption,and disappearance along with lymphatic vessel infiltration in bone-marrow cavities.Although the osteolytic mechanism of GSD has been widely studied,the cause of lymphatic hyperplasia in GSD is rarely investigated.In this study,by comparing the RNA expression profile of osteoclasts(OCs)with that of OC precursors(OCPs)by RNA sequencing,we identified a new factor,semaphorin 3A(Sema3A),which is an osteoprotective factor involved in the lymphatic expansion of GSD.Compared to OCPs,OCs enhanced the growth,migration,and tube formation of lymphatic endothelial cells(LECs),in which the expression of Sema3A is low compared to that in OCPs.In the presence of recombinant Sema3A,the growth,migration,and tube formation of LECs were inhibited,further confirming the inhibitory effect of Sema3A on LECs in vitro.Using an LEC-induced GSD mouse model,the effect of Sema3A was examined by injecting lentivirus-expressing Sema3A into the tibiae in vivo.We found that the overexpression of Sema3A in tibiae suppressed the expansion of LECs and alleviated bone loss,whereas the injection of lentivirus expressing Sema3A short hairpin RNA(shRNA)into the tibiae caused GSD-like phenotypes.Histological staining further demonstrated that OCs decreased and osteocalcin increased after Sema3A lentiviral treatment,compared with the control.Based on the above results,we propose that reduced Sema3A in OCs is one of the mechanisms contributing to the pathogeneses of GSD and that expressing Sema3A represents a new approach for the treatment of GSD.

AIM: To investigate the effect of electroacupuncture on the expression of matrix metalloproteinase-3(MMP-3), tissue inhibitor of metalloproteinase-3(TIMP-3)and collagen type III alpha 1(Col3α1)in the retina of guinea pigs with lens-induced myopia.METHODS:A total of 80 guinea pigs were randomly divided into the normal control(NC)group, negative lens-induced myopia(LIM)group, electro-acupuncture(EA)group, and sham group, with 20 guinea pigs in each group. The NC group did not receive any intervention, the LIM, EA and the sham groups were all wearing -6.0 D lens in the right eye and no lens in the left eye. The EA group was given electroacupuncture stimulation at Hegu Point and temple, and guinea pigs in the sham group were given intervention at false points. The refraction was measured by optometry, the axial length was measured by A-ultrasound, and the changes in retinal tissue structure were observed by HE staining. Quantitative polymerase chain reaction(Q-PCR)and western blot(WB)were used to detect the expression of MMP-3, TIMP-3, and Col3α1 at mRNA and protein levels before modeling and at 2 and 4 wk after modeling. RESULTS:The axial length of the negative LIM group was significantly increased compared with the NC group at 2 and 4 wk after modeling(all P<0.05), and the diopter was significantly decreased(all P<0.05); the axial length of the eye in the EA group was decreased compared with the negative LIM group(all P<0.05), and the diopter was increased(all P<0.05). HE staining showed that the boundaries of retinal tissue in the NC group were obvious and arranged regularly. The retinal thickness, the thickness of the inner and outer nuclear layers, and the number of cells were reduced and irregularly arranged in the negative LIM group. The overall structure of the retina in the group EA was relatively perfect, the arrangement was more regular, and the morphological structure of each layer of the tissue did not appear obvious abnormalities. Q-PCR and WB detection results showed that the mRNA and protein relative expression levels of MMP-3, TIMP-3, and Col3α1 in the negative LIM group were significantly higher than those in the NC group(all P<0.05); the expression levels of the EA group was significantly decreased after intervention compared with the negative LIM group(all P<0.05).CONCLUSION: Electroacupuncture can delay axial growth in negative LIM guinea pigs, and downregulate negative LIM induced expression of MMP-3, TIMP-3, and Col3α1 in the retina.