BACKGROUND: Chronic kidney disease (CKD) is associated with common pathophysiological processes, such as inflammation and fibrosis, in both the heart and the kidney. However, the underlying molecular mechanisms that drive these processes are not yet fully understood. Therefore, this study focused on the molecular mechanism of heart and kidney injury in CKD. METHODS: We generated an microRNA (miR)-26a knockout (KO) mouse model to investigate the role of miR-26a in angiotensin (Ang)-II-induced cardiac and renal injury. We performed Ang-II modeling in wild type (WT) mice and miR-26a KO mice, with six mice in each group. In addition, Ang-II-treated AC16 cells and HK2 cells were used as in vitro models of cardiac and renal injury in the context of CKD. Histological staining, immunohistochemistry, quantitative real-time polymerase chain reaction (PCR), and Western blotting were applied to study the regulation of miR-26a on Ang-II-induced cardiac and renal injury. Immunofluorescence reporter assays were used to detect downstream genes of miR-26a, and immunoprecipitation was employed to identify the interacting protein of LIM and senescent cell antigen-like domain 1 (LIMS1). We also used an adeno-associated virus (AAV) to supplement LIMS1 and explored the specific regulatory mechanism of miR-26a on Ang-II-induced cardiac and renal injury. Dunnett's multiple comparison and t -test were used to analyze the data. RESULTS: Compared with the control mice, miR-26a expression was significantly downregulated in both the kidney and the heart after Ang-II infusion. Our study identified LIMS1 as a novel target gene of miR-26a in both heart and kidney tissues. Downregulation of miR-26a activated the LIMS1/integrin-linked kinase (ILK) signaling pathway in the heart and kidney, which represents a common molecular mechanism underlying inflammation and fibrosis in heart and kidney tissues during CKD. Furthermore, knockout of miR-26a worsened inflammation and fibrosis in the heart and kidney by inhibiting the LIMS1/ILK signaling pathway; on the contrary, supplementation with exogenous miR-26a reversed all these changes. CONCLUSIONS Our findings suggest that miR-26a could be a promising therapeutic target for the treatment of cardiorenal injury in CKD. This is attributed to its ability to regulate the LIMS1/ILK signaling pathway, which represents a common molecular mechanism in both heart and kidney tissues.
Animals ; MicroRNAs/metabolism* ; Angiotensin II/toxicity* ; Mice ; Renal Insufficiency, Chronic/chemically induced* ; Mice, Knockout ; Disease Models, Animal ; Male ; Signal Transduction/genetics* ; LIM Domain Proteins/genetics* ; Mice, Inbred C57BL ; Cell Line ; Humans

[Objective] To establish a cryopreservation protocol for small-volume (≤1 mL) red blood cells (RBCs) and to evaluate the reactivity and stability of blood group antigens after cryopreservation, so as to explore its potential application in immunohematology reference laboratories. [Methods] Small-volume RBCs were cryopreserved for 120 days, followed by thawing and deglycerolization to restore the RBC components. The quality of the RBCs was assessed. Serum antibodies were serially diluted and reacted with RBCs before and after cryopreservation, and agglutination scores were recorded to quantitatively evaluate the reactivity and stability of blood group antigens such as Rh, Duffy, Lewis, Kidd, M, and H. Flow cytometry was used to analyze the percentage and mean fluorescence intensity of ABO antigen expression on RBCs before and after cryopreservation to assess the usability of cryopreserved RBCs in flow immunophenotyping and blood group subtype studies. [Results] The hemolysis rate of thawed and deglycerolized RBCs was (0.27±0.10)%, with a supernatant free hemoglobin level of (0.52±0.14) g/L, and the RBC recovery rate was (69.12±7.91)%. The direct antiglobulin test (DAT) was negative for all thawed RBCs. There was no difference in the reactivity of blood group antigens before and after cryopreservation, and no difference in the percentage and mean fluorescence intensity of A and B antigen expression on RBCs before and after cryopreservation. [Conclusion] The small-volume RBC cryopreservation protocol can be applied to immunohematology analysis in reference laboratories and is expected to be widely used in blood group identification, antibody screening, identification, and blood group-related research.

ObjectiveTo investigate the application of the novel inflammatory factor adsorption column CA280 combined with low-dose plasma exchange （LPE） in patients with acute-on-chronic liver failure （ACLF）. MethodsA prospective cohort study was designed， and a total of 93 ACLF patients who were admitted to The Ninth Hospital of Nanchang from June 2023 to January 2025 were enrolled and randomly divided into DPMAS+LPE group with 50 patients and CA280+LPE group with 43 patients. In addition to comprehensive medical treatment， the patients in the DPMAS+LPE group received DPMAS and LPE treatment， and those in the CA280+LPE group received CA280 and LPE treatment. The two groups were observed in terms of routine blood test results， liver function parameters， renal function markers， electrolytes， coagulation function parameters， cytokines， adverse events， and 28-day prognosis before surgery （baseline）， during surgery （DPMAS or CA280）， and after surgery （after sequential LPE treatment）. The paired t-test was used for comparison of normally distributed continuous data before and after treatment within each group， and the independent-samples t test was used for comparison between groups； the Wilcoxon signed-rank test was used for comparison of non-normally distributed continuous data before and after treatment within each group， and the Mann-Whitney U test was used for comparison between groups. The chi-square test or the Fisher’s exact test was used for comparison of categorical data between groups， and the Spearman test was used for correlation analysis. ResultsAfter CA280 treatment， the ACLF patients had significant reductions in the levels of cytokines （IL-6， IL-8， IL-10， TNF-α， and IFN-γ）， liver function parameters （ALT， AST， ALP， TBil， DBil， Alb， and glutathione reductase）， and the renal function marker urea nitrogen （all P<0.05）， and in terms of coagulation function parameters， there were significant increases in prothrombin time， activated partial thromboplastin time （APTT）， thrombin time， and international normalized ratio （INR） and significant reductions in prothrombin activity （PTA） and fibrinogen （FIB） （all P<0.05）. Compared with the DPMAS+LPE group， the CA280+LPE group showed better improvements in the serum cytokines IL-8 （Z=-2.63， P=0.009）， IL-10 （Z=-3.94， P<0.001）， and TNF-α （Z=-1.53， P=0.023）， and the two artificial liver support systems had a similar effect in improving liver function （ALT， AST， GGT， GR， TBil， and DBil） （all P >0.05）， but the CA280+LPE group showed a significantly greater reduction in Alb （Z=-2.08， P=0.037）. CA280+LPE was more effective in reducing uric acid （Z=-2.97， P=0.003）. Compared with DPMAS+LPE， CA280+LPE treatment resulted in a significant reduction in INR （Z=-4.01， P<0.001）， a significant increase in APTT （Z=-2.53， P=0.011）， and significant greater increases in PTA （Z=-6.28， P<0.001） and FIB （Z=-3.93， P<0.001）. There were no significant differences in the incidence rates of adverse reactions and the rate of improvement at discharge between the two groups （all P>0.05）. The Spearman correlation analysis showed that IL-6 was significantly correlated with WBC （r=0.22， P=0.042）， TBil （r=0.29， P=0.005）， and FIB （r=-0.33， P=0.003）； IL-8 was positively correlated with APTT （r=0.37， P<0.001） and INR （r=0.25， P=0.013）； TNF-α was significantly correlated with WBC （r=0.40， P<0.001） and TBil （r=0.34， P<0.001）. ConclusionCompared with DPMAS， CA280 combined with LPE can effectively clear proinflammatory cytokines and improve liver function in ACLF patients， but it has a certain impact on Alb and coagulation function. This regimen provides a new option for the individualized treatment of ACLF and can improve the short-term prognosis of patients， but further studies are needed to verify its long-term efficacy.

Objective To investigate the clinical features of thyroid dysfunction in lung cancer patients treated with programmed cell death receptor-1(PD-1)or programmed cell death receptor-ligand 1(PD-L1)and their value for predicting therapeutic efficacy.Methods Lung cancer patients treated with PD-1/PD-L1 inhibitors at West China Hospital,Sichuan University between March 2018 and September 2022 were retrospectively enrolled.Data concerning the medical records,therapeutic efficacy,and thyroid function indicators of the patients were retrieved from the hospital electronic medical record information system.The data were then analyzed to identify risk factors and predictive factors for immune-related adverse events(irAEs)of the thyroid.The predictive value of thyroid irAEs for treatment efficacy and prognosis was assessed.Objective response rate(ORR)was defined as the indicator for therapeutic efficacy and progression-free survival(PFS)was defined as the prognostic indicator.Results A total of 368 lung cancer patients were enrolled.Among them,31.5％(116/368)developed thyroid irAEs.According to the results of logistic regression analysis,baseline thyroid stimulating hormone(TSH)concentration and baseline positive results for thyroglobulin antibody(TGAb)and thyroid peroxidase antibody(TPOAb)were risk factors for thyroid dysfunction caused by PD-1/PD-L1 inhibitors.Among the three measures,baseline TPOAb concentration demonstrated good predictive value for thyroid irAEs,with an area under the receiver-operating characteristic(ROC)curve(AUC)of 0.745.Patients with thyroid irAEs had a longer median PFS(16.0 months vs.9.7 months,P＜0.001)and a higher ORR(55.2％vs.34.9％,P＜0.001)compared to those without thyroid irAEs.Patients with thyroid irAEs had a better ORR than those without thyroid irAEs did.It was more likely for patients with thyroid irAEs to achieve an objective response compared to those without thyroid irAEs(odds ratio[OR]=2.29;95％CI,1.46-3.60).Conclusion In lung cancer patients treated with the PD-1/PD-L1 inhibitors,the TPOAb antibody demonstrates good predictive value for thyroid irAEs.Patients who develop thyroid irAEs have better treatment outcomes and prognosis.

Objective:To investigate the differences in protein profile expression in serum samples from children with corona virus disease 2019（COVID-19）related encephalopathy and to explore the underlying mechanisms.Methods:From December 1,2022 to January 31,2023,28 children with COVID-19 who were admitted to the Department of Pediatric Intensive Medicine at Shandong Provincial Hospital Affiliated to Shandong First Medical University were collected,including 21 patients with encephalopathy(COVID-19 with encephalopathy group) and seven patients without encephalopathy(COVID-19 without encephalopathy group).Three children from each group were selected for serum proteomic analysis using tandem mass spectrometry labeling proteomics technology.Proteins were considered significantly different if the fold change was >1.2 or <0.8，with P<0.05.Bioinformatics analysis，including Gene Ontology and Kyoto Encyclopedia of Genes and Genomes Pathway Enrichment were performed on differentially expressed proteins.Protein-protein interaction networks were analyzed using the STRING database.Selected proteins were further validated by enzyme-linked immunosorbert assay. Results:A total of 41 differentially expressed proteins were identified between the two groups.Among these，14 proteins were upregulated and 27 proteins were downregulated in COVID-19 patients with encephalopathy compared to those without encephalopathy.Bioinformatics analysis revealed that these proteins were primarily enriched in critical signaling pathways，including complement and coagulation regulation，neutrophil degranulation and activation，and platelet degranulation.Enzyme-linked immunosorbert assay validation confirmed significant differences in key coagulation-regulating proteins(von willebrand factor upregulated，serpin family F member 2 downregulated in COVID-19 patients with encephalopatly）between the two groups.Conclusion:Coagulation dysfunction may play a role in the development of COVID-19 associated encephalopathy in children，providing valuable insights for future research.

OBJECTIVE To observe the effect of Shixiang plaster on promoting the wound healing of diabetic foot ulcer.METHODS Totally 50 male SPF grade SD rats were prepared to establish the diabetic models by feeding with high glucose and high fat forage and intraperitoneal injection of streptozotocin,the rats models that were es-tablished successfully were randomly divided into the model group,the Shixiang plaster group and the Kangfuxin solution group.The models of diabetic ulcers were established.The Shixiang plaster group was treated with exter-nal Shixiang plaster,the Kangfuxin solution group was given external Kangfuxin solution,and the model group was treated with coverage with sterile gauze.The wound healing status of the rats was observed,the wound tis-sues were collected for bacterial culture and hematoxylin-eosin(HE)staining after drug administration for 14 and 28 days,respectively.The expression levels of nuclear factor-red cell-2-related factor 2(Nrf-2),heme oxygenase-1(HO-1),tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and vascular endothelial growth factor(VEGF)were detected by immunohistochemistry(IHC),the expression levels of Nrf-2 and HO-1 were detected with the use of Western Blot,and the levels of serum malondialdehyde(MDA),superoxide dismutase(SOD)and reactive oxygen species(ROS)were detected by enzyme-linked immunosorbent assay(ELISA).RESULTS After the drug administration for 14 and 28 days,the wound healing rates of the Shixiang plaster group were(62.15±3.82)％and(81.68±3.83)％,respectively,higher than(47.14±2.80)％and(69.96±6.49)％of the Kangfuxin solu-tion group and(29.14±9.52)％and(57.91±6.63)％of the model group,and there were significant differences(F=21.716,12.626,P=0.002,0.007).The bacterial colony counts of the Shixiang plaster group were less than those of the Kangfuxin solution group and the model group after the drug administration for 14 and 28 days(P＜0.05).The result of HE staining showed that the Shixiang plaster group had a better wound healing.The result of IHC indicated that the expression levels of Nrf-2,HO-1 and VEGF of the Kangfuxin solution group and the Shix-iang plaster group were up-regulated after the drug administration for 28 days,while the expression levels of TNF-α and IL-6 were down-regulated.The expression levels of Nrf-2 and HO-1 proteins of the Shixiang plaster group were higher than those of the Kangfuxin solution group and the model group after the drug administration for 14 and 28 days,the levels of serum MDA and ROS of the Shixiang plaster group were lower than those of the Kangfuxin solution group and the model group,and the serum SOD level of the Shixiang plaster group was higher than that of the model group(P＜0.05).CONCLUSIONS Shixiang plaster can effectively promote the wound heal-ing of the rats with diabetic foot ulcers and reduce the bacterial colony counts of wound surfaces.The mechanism may be associated with the alleviation of oxidative stress injury by mediating the Nrf-2/HO-1 signaling pathways,promotion of angiogenesis and inhibition of excessive inflammatory reactions.

OBJECTIVE To observe the effect of Shixiang plaster on promoting the wound healing of diabetic foot ulcer.METHODS Totally 50 male SPF grade SD rats were prepared to establish the diabetic models by feeding with high glucose and high fat forage and intraperitoneal injection of streptozotocin,the rats models that were es-tablished successfully were randomly divided into the model group,the Shixiang plaster group and the Kangfuxin solution group.The models of diabetic ulcers were established.The Shixiang plaster group was treated with exter-nal Shixiang plaster,the Kangfuxin solution group was given external Kangfuxin solution,and the model group was treated with coverage with sterile gauze.The wound healing status of the rats was observed,the wound tis-sues were collected for bacterial culture and hematoxylin-eosin(HE)staining after drug administration for 14 and 28 days,respectively.The expression levels of nuclear factor-red cell-2-related factor 2(Nrf-2),heme oxygenase-1(HO-1),tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and vascular endothelial growth factor(VEGF)were detected by immunohistochemistry(IHC),the expression levels of Nrf-2 and HO-1 were detected with the use of Western Blot,and the levels of serum malondialdehyde(MDA),superoxide dismutase(SOD)and reactive oxygen species(ROS)were detected by enzyme-linked immunosorbent assay(ELISA).RESULTS After the drug administration for 14 and 28 days,the wound healing rates of the Shixiang plaster group were(62.15±3.82)％and(81.68±3.83)％,respectively,higher than(47.14±2.80)％and(69.96±6.49)％of the Kangfuxin solu-tion group and(29.14±9.52)％and(57.91±6.63)％of the model group,and there were significant differences(F=21.716,12.626,P=0.002,0.007).The bacterial colony counts of the Shixiang plaster group were less than those of the Kangfuxin solution group and the model group after the drug administration for 14 and 28 days(P＜0.05).The result of HE staining showed that the Shixiang plaster group had a better wound healing.The result of IHC indicated that the expression levels of Nrf-2,HO-1 and VEGF of the Kangfuxin solution group and the Shix-iang plaster group were up-regulated after the drug administration for 28 days,while the expression levels of TNF-α and IL-6 were down-regulated.The expression levels of Nrf-2 and HO-1 proteins of the Shixiang plaster group were higher than those of the Kangfuxin solution group and the model group after the drug administration for 14 and 28 days,the levels of serum MDA and ROS of the Shixiang plaster group were lower than those of the Kangfuxin solution group and the model group,and the serum SOD level of the Shixiang plaster group was higher than that of the model group(P＜0.05).CONCLUSIONS Shixiang plaster can effectively promote the wound heal-ing of the rats with diabetic foot ulcers and reduce the bacterial colony counts of wound surfaces.The mechanism may be associated with the alleviation of oxidative stress injury by mediating the Nrf-2/HO-1 signaling pathways,promotion of angiogenesis and inhibition of excessive inflammatory reactions.

Objective:To investigate the differences in protein profile expression in serum samples from children with corona virus disease 2019（COVID-19）related encephalopathy and to explore the underlying mechanisms.Methods:From December 1,2022 to January 31,2023,28 children with COVID-19 who were admitted to the Department of Pediatric Intensive Medicine at Shandong Provincial Hospital Affiliated to Shandong First Medical University were collected,including 21 patients with encephalopathy(COVID-19 with encephalopathy group) and seven patients without encephalopathy(COVID-19 without encephalopathy group).Three children from each group were selected for serum proteomic analysis using tandem mass spectrometry labeling proteomics technology.Proteins were considered significantly different if the fold change was >1.2 or <0.8，with P<0.05.Bioinformatics analysis，including Gene Ontology and Kyoto Encyclopedia of Genes and Genomes Pathway Enrichment were performed on differentially expressed proteins.Protein-protein interaction networks were analyzed using the STRING database.Selected proteins were further validated by enzyme-linked immunosorbert assay. Results:A total of 41 differentially expressed proteins were identified between the two groups.Among these，14 proteins were upregulated and 27 proteins were downregulated in COVID-19 patients with encephalopathy compared to those without encephalopathy.Bioinformatics analysis revealed that these proteins were primarily enriched in critical signaling pathways，including complement and coagulation regulation，neutrophil degranulation and activation，and platelet degranulation.Enzyme-linked immunosorbert assay validation confirmed significant differences in key coagulation-regulating proteins(von willebrand factor upregulated，serpin family F member 2 downregulated in COVID-19 patients with encephalopatly）between the two groups.Conclusion:Coagulation dysfunction may play a role in the development of COVID-19 associated encephalopathy in children，providing valuable insights for future research.

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.

The patient,a male newborn,was admitted to the hospital 2 hours after birth due to prematurity(gestational age 27+5 weeks)and respiratory distress occurring 2 hours postnatally.After admission,the infant developed fever and elevated C-reactive protein levels.On the fourth day after birth,metagenomic next-generation sequencing of cerebrospinal fluid indicated a positive result for Mycoplasma hominis(9 898 reads).On the eighth day,a retest of cerebrospinal fluid metagenomics confirmed Mycoplasma hominis(56 806 reads).The diagnosis of purulent meningitis caused by Mycoplasma hominis was established,and the antibiotic treatment was switched to moxifloxacin[5 mg/(kg·day)]administered intravenously for a total of 4 weeks.After treatment,the patient's cerebrospinal fluid tests returned to normal,and he was discharged as cured on the 76th day after birth.This article focuses on the diagnosis and treatment of neonatal Mycoplasma hominis purulent meningitis,introducing the multidisciplinary diagnosis and treatment of the condition in extremely preterm infants.[Chinese Journal of Contemporary Pediatrics,2024,26(4):432-436]