AIM: To investigate the changes of retinal nerve fiber layer(RNFL)thickness, retinal thickness and blood flow density in different stages of diabetic retinopathy(DR)patients based on optical coherence tomography angiography(OCTA).METHODS: A retrospective analysis was conducted on 382 patients(382 eyes)diagnosed with DR in our hospital from February 2023 to February 2024. According to the staging criteria, the patients were divided into mild group(n=121), moderate group(n=133), severe group(n=72), and proliferative group(n=56). The general clinical data of the four groups of patients was compared; OCTA was used to scan and collect data from all patients, and the RNFL thickness, retinal thickness, and blood flow density were compared among the four groups of patients.RESULTS: There was no statistically significant difference in age, gender, hypertension, chronic kidney disease, and random blood glucose among patients in the mild, moderate, severe, and proliferative groups(all P>0.05). As the stage of DR worsened, the duration of the disease gradually prolonged(P<0.05). The thickness of the RNFL(superior, inferior, temporal, nasal, and average thickness)and retinal thickness significantly increased with the severity of DR(all P<0.001); however, there was no statistically significant difference in inferior RNFL thickness between the moderate and mild groups(P>0.05). The blood flow density in the superficial and deep retinal layers, as well as in the choroidal capillary layer, significantly decreased with the progression of DR(all P<0.05). Nevertheless, there was no statistically significant difference in superficial retinal blood flow density between the moderate and severe groups(P>0.05).CONCLUSION: OCTA can accurately observe the changes in RNFL thickness, retinal thickness, and blood flow density in patients with DR at different stages, which can serve as sensitive indicators for monitoring DR progression.

Objective: To compare the changes in the concentration of relevant growth factors released from platelet-rich plasma (PRP) stored at -80℃ by cryopreservation and at 4℃ by refrigerated lyophilization over 2 years, aiming to provide a theoretical basis for prolonging PRP storage duration. Methods: PRP (n=15) was separated using a blood cell separator and stored under -80℃ cryopreservation (F-PRP group) and 4℃ refrigerated freeze-drying conditions (FD-PRP group). The contents of growth factors (PDGF-AA, PDGF-BB, EGF, TGF-β1, and VEGF) in both groups were measured by ELISA at 1, 3, 6, 9, 12 and 24 months. Results: PDGF-AA and VEGF maintained good stability in both groups for up to 24 months. PDGF-BB and TGF-β1 showed high stability in the first 12 months but their stability decreased gradually from 12th to 24th months. EGF demonstrated good stability in the first 6 months, and its stability gradually decreased from the 9th to 24th months. Comparing the F-PRP and FD-PRP groups, the concentrations of the five growth factors in the FD-PRP group were either not statistically different or higher than those in the F-PRP group at all time points. Specifically, the concentrations of EGF were significantly higher in the FD-PRP group at all time points. Conclusion: Both -80℃ freezing and 4℃ freeze-drying enable long-term preservation of PRP. Freeze-drying imposes less stringent storage requirements and facilitates growth factor compared to frozen storage.

Objective To investigate the role and mechanism of RNA-Specific adenosine deaminase 3 (Adar3) in regulating macrophage polarization during Coxsackievirus B3(CVB3)-induced viral myocarditis (VM). Methods Bone marrow-derived macrophages (BMDM) from mice were cultured in vitro and induced into M1/M2 macrophages using interferon-gamma (IFN-γ)/lipopolysaccharide (LPS) or interleukin 4 (IL-4), respectively. The mRNA expression levels of Adar1, Adar2, and Adar3 in each group of cells were assessed by real-time quantitative PCR (qRT-PCR). Specific siRNAs targeting the Adar3 gene were designed, synthesized, and transiently transfected into M2 macrophages. The mRNA levels of M2 polarization-related marker genes-including arginase 1 (Arg1), chitinase 3-like molecule 3 (YM1/Chi3l3), and resistin-like molecule alpha (RELMα/FIZZ1)-were detected by qRT-PCR. RNA sequencing was performed to analyze the signaling pathways affected by Adar3. The expression levels of Wnt/β-catenin signaling pathway were further validated using qRT-PCR and Western blot. The adeno-associated virus overexpressing Adar3 was designed, synthesized, and injected into mice via tail vein. Three weeks later, a myocarditis mouse model was established. After an additional week, the phenotype and function of cardiac macrophages, as well as multiple indicators of VM (including echocardiography, body weight, histopathology and serology) were examined. Additionally, the protein levels of the Wnt/β-catenin signaling pathway were assessed. Results Compared to M0-type macrophages, the expression level of Adar3 was significantly increased in M2-type macrophages. After transfection of Adar3 siRNA, the mRNA levels of Arg1, YM1 and FIZZ1 in M2 macrophages were downregulated. RNA sequencing revealed 149 upregulated genes and 349 downregulated genes. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis and subsequent validation experiments indicated that Adar3 modulated the Wnt/β-catenin signaling pathway. In vivo experiments demonstrated that Adar3 overexpression alleviated the cardiac dysfunction of VM mice. The proportion of M1 macrophages in the heart decreased, while the proportion of M2 macrophages increased. At the same time, the Adar3 overexpression activated the Wnt/β-catenin signaling pathway. Conclusion Adar3 promotes macrophage polarization toward the M2 phenotype by activating the Wnt/β-catenin signaling pathway, thereby alleviating VM.
Animals ; Adenosine Deaminase/metabolism* ; Macrophages/immunology* ; Wnt Signaling Pathway/genetics* ; Myocarditis/immunology* ; Mice ; Coxsackievirus Infections/metabolism* ; Male ; Mice, Inbred BALB C ; Enterovirus B, Human/physiology* ; beta Catenin/genetics*

Objective:This aims to investigate the diagnostic and evaluative value of MRI for lymphatic malformations in the head, neck, and facial regions of children. Methods:A retrospective analysis was conducted on the MRI imaging data of 31 cases of head, neck, and facial lymphatic malformations in children admitted to the Department of Otolaryngology, Head and Neck Surgery, Children's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, from January 2022 to January 2024. Results:The MRI images of this group of cases primarily displayed irregular morphology（80.6%, 25/31）, thin-walled cysts（80.6%, 25/31）, and compression of surrounding tissues. The boundaries were clear（100%, 31/31）, with characteristics of invasive and drill-like growth（93.5%）. The cyst walls or internal septa exhibited high signal intensity on T1WI, low signal intensity on T2WI, and mild to moderate enhancement（100%）. The contents of the cysts showed low signal intensity on T1WI, high signal intensity on T2WI, and no enhancement（35.5%, 11/31）. Mixed signals with varying degrees of enhancement were observed in 20 cases（64.5%）. There were 29 cases of multilocular cysts（93.5%, 29/31）, and 11 cases of fluid-fluid levels（35.5%）. The MRI diagnostic accuracy for this group of cases was 100%. Conclusion:Lymphatic Malformations of head, neck and facial region in children have very characteristic features on MRI, such as typical thin wall, clear boundaries, irregular shapes, invasive growth, no enhancement, multilocular cystic masses, fluid-fluid level, etc. Furthermore, it is more appropriate for children with lymphatic malformations owing to its non-radiation and non-invasive benefits. Diagnosing lymphatic malformations in the head, neck, and facial region in children should begin with this.
Humans ; Retrospective Studies ; Lymphatic Abnormalities/diagnostic imaging* ; Magnetic Resonance Imaging ; Neck/diagnostic imaging* ; Head/diagnostic imaging* ; Face/diagnostic imaging* ; Child ; Male ; Female ; Child, Preschool ; Adolescent ; Infant

Mitochondrial DNA (mtDNA) acts as a damage-associated molecular pattern to activate the stimulator of interferon genes (STING) signaling in macrophages, promoting tissue inflammation. However, its role in acute myocardial infarction (AMI) remains unclear. Macrophage-specific Sting1 knockout mice were used to validate STING's pathological role in AMI. Cardiac and liver mtDNA were used to activate macrophages in co-culture systems with cardiomyocytes to assess fibrosis and hypertrophy. Panaxatriol saponin (PTS) was tested for its ability to block mtDNA-driven macrophage activation and subsequent cardiomyocyte damage. STING-PTS binding ability was analyzed. AMI rats received PTS to evaluate its effects on myocardial inflammation and ventricular remodeling. In vivo, macrophage-specific Sting1 knockout reduced myocardial inflammation and injury after AMI. In vitro, mtDNA-activated macrophages induced cardiomyocyte fibrosis and hypertrophy through STING signaling. PTS suppressed mtDNA-driven macrophage activation by directly binding STING, thereby blocking inflammatory cascades. In AMI rats, PTS treatment attenuated acute inflammation and reversed ventricular remodeling. These findings establish the mtDNA-STING axis in macrophages as a critical driver of post-AMI inflammation and identify pharmacological STING inhibition with PTS as a promising therapeutic strategy. The study bridges genetic validation with translational applications, highlighting macrophage STING as a novel target for ischemic heart disease management.

Objective To understand the internal quality control(IQC)situation of routine biochemical projects in Guizhou Province from 2020 to 2022,and to use the analysis batch length sigma rule to select the appropriate IQC strategy for routine biochemical projects.Methods A sum of 41,41 and 52 laboratories were selected from the laboratories participating in the first external quality assessment(EQA)program of the routine biochemical project in the Guizhou Provincial Center for Clinical Laboratory from 2020 to 2022,which uniformly used two concentration levels of internal quality control products produced by Randox Company in the UK.The coefficient of variation(CV)was calculated based on their IQC information,and then these CV values were used to compare with imprecise performance specifications such as WS/T403-2012 standards.The IQC situation of routine biochemical projects of Guizhou Province in recent years was analyzed.Randomly 15 laboratories were selected each year to report the results of the triglyceride project,and their σ values were calculated.Based on the Westgard sigma rule flowcharts of the analysis batch length,appropriate quality control strategies were selected and the quality goal index(QGI)was calculated to evaluate analytical performance of laboratory detection system.Results In the current monthly CV,the proportion of laboratories that met the WS/T403-2012 standards for Ca and TP projects was relatively low,while the pass rates for 7 projects(P,CK,LDH and other projects)were all above 80%,showing an increasing trend year by year.The proportion of laboratories with the 7 projects(P,CK,LDH and other projects)meeting the performance specifications for biomedical variation(appropriate)all reached 90%,while the proportion of laboratories with 5 projects(Na,Ca,Cl and other projects)was relatively low.The proportion of 10 projects such as P,CK,and LDH in cumulative CV of laboratories that met the performance specifications for biomedical variation(appropriate)was over 90%.For the triglyceride project,from 2020 to 2022,there were 2,5 and 13 laboratories that reached 6 σ horizontal.Compared to values of 2020 and 2021 years,the trend of σ values changes of 2022 was increasing(t=3.855,3.511,P≤0.001),indicating that the analytical performance of these laboratories was getting better and better.Conclusion The IQC levels of routine biochemical projects in Guizhou Province were increasing year by year.The Westgard sigma rule of the analysis batch length is designed with personalized quality control rules for various laboratories in Guizhou Province,thus improving laboratory testing capabilities and better severing clinics.

BACKGROUND:Tissue engineering has brought new hope to the clinical challenge of liver failure,and the preparation of plant-derived decellularized fiber scaffolds holds significant importance in liver tissue engineering. OBJECTIVE:To prepare apple tissue decellularized scaffold material by using fresh apple slices and a solution of sodium dodecyl sulfate,and assess its biocompatibility. METHODS:Fresh apples were subjected to decellularization using phosphate buffer saline and sodium dodecyl sulfate solution,separately.Afterwards,the decellularized apple tissues and apple decellularized scaffold materials were decontaminated with phosphate buffer saline.Subsequently,scanning electron microscopy was used to assess the effectiveness of decellularization of the apple materials.Adipose-derived mesenchymal stem cells were extracted from the inguinal fat BALB/C of mice,and their expression of stem cell-related markers(CD45,CD34,CD73,CD90,and CD105)was identified through flow cytometry.The cells were then divided into a scaffold-free control group and a scaffold group.Equal amounts of adipose-derived mesenchymal stem cells were seeded onto both groups.The biocompatibility of the decellularized scaffold with adipose-derived mesenchymal stem cells was evaluated using CCK-8 assay,hematoxylin-eosin staining,and phalloidine staining.Cell adhesion and growth on the scaffold were observed under light microscopy and scanning electron microscopy.Furthermore,the scaffold was subdivided into the non-induced group and the hepatogenic-induced group.Adipose-derived mesenchymal stem cells were cultured on the decellularized apple scaffold,and they were cultured for 14 days in regular culture medium or hepatogenic induction medium for comparison.Immunofluorescent staining using liver cell markers,including albumin,cytokeratin 18,and CYP1A1,was performed.Enzyme-linked immunosorbent assay was used to detect the secretion of alpha fetoprotein and albumin.Additionally,scanning electron microscopy was employed to observe the morphology of the induced cells on the scaffold,verifying the expression of liver cell-related genes on the decellularized scaffold material.Finally,the cobalt-60 irradiated and sterilized decellularized apple scaffolds were transplanted onto the surface of mouse liver and the degradation of the scaffold was observed by gross observation and hematoxylin-eosin staining after 28 days. RESULTS AND CONCLUSION:(1)The scanning electron microscopy results revealed that the decellularized apple scaffold material retained a porous structure of approximately 100 μm in size,with no residual cells observed.(2)Through flow cytometry analysis,the cultured cells were identified as adipose-derived mesenchymal stem cells.(3)CCK-8 assay results demonstrated that the prepared decellularized apple tissue scaffold material exhibited no cytotoxicity.Hematoxylin-eosin staining and phalloidine staining showed that adipose-derived mesenchymal stem cells were capable of adhering and proliferating on the decellularized apple tissue scaffold.(4)The results obtained from immunofluorescence staining and enzyme-linked immunosorbent assay revealed that adipose-derived mesenchymal stem cells cultured on the decellularized apple scaffolds exhibited elevated expression of liver-specific proteins,including albumin,alpha-fetoprotein,cytokeratin 18,and CYP1A1.These results suggested that they were induced differentiation into hepatocyte-like cells possessing functional characteristics of liver cells.(5)The decellularized apple scaffold implanted at 7 days has integrated with the liver,with partial degradation of the scaffold observed.By 28 days,the decellularized apple scaffold has completely degraded and has been replaced by newly-formed tissue.(6)The results indicate that the decellularized scaffold material derived from apple tissue demonstrates favorable biocompatibility,promoting the proliferation,adhesion,and hepatic differentiation of adipose-derived mesenchymal stem cells.

Objective To analyze the distribution characteristics of traditional Chinese medicine(TCM)syndrome types in patients with osteoporosis and the distribution differences of clinical and serological indicators in TCM syndrome types.Meth-ods A total of 69 patients with osteoporosis were collected from the Third Affiliated Hospital of Guangzhou University of Chinese Medicine and Qifu Hospital Affiliated to Jinan University.The general information,bone mineral density T value,fasting periph-eral venous blood in the morning were collected.The expression of telomerase protective factor 1(POT1),telomerase reverse transcriptase(TERT),serum 8-hydroxy-2'-deoxyguanosine(8-OHdG)and superoxide dismutase 2(SOD2)were detected by ELISA.Finally,the above data were statistically analyzed.Results There were significant differences in body weight,height,bone mineral density,POT1,TERT,and 8-OHdG among the four syndromes(P＜0.05).In terms of correlation,the relation-ship between bone mineral density and each parameter in different syndrome types was explored.The bone mineral density of qi stagnation and blood stasis syndrome was positively correlated with SOD2 value.There is a positive correlation between bone min-eral density and 8-OHdG in patients with Yin deficiency of liver and kidney.TERT was positively correlated with qi stagnation and blood stasis syndrome.Liver and kidney Yin deficiency syndrome was positively correlated with weight and bone mineral den-sity,and negatively correlated with TERT value.Weight was negatively correlated with qi and blood stasis syndrome.Conclusion In TCM syndrome differentiation of osteoporosis,there were statistical differences in weight,height,bone mineral density,ser-um POT1,TERT and 8-OHdG among Qi-stagnation and blood stasis,spleen-kidney Yang deficiency,liver-kidney Yin deficiency and Qi-blood-peace syndrome.In different syndrome types,serum SOD2 and 8-OHdG were the influencing factors of bone miner-al density.Serum TERT and 8-OHdG are the main factors affecting the dialectical classification of osteoporosis.

Although a great deal of research has been done on sepsis in children，its pathogenesis remains controversial and unresolved，one of which is immune dysfunction.Immunosuppression is the core part of immune dysfunction，and it is related to the disease progression，treatment options，and prognosis changes in patients with sepsis.Since T lymphocytes play a crucial role in immune function，they have been regarded as an important indicator for judging changes in immune function in clinical practice.This review explained the mechanism of immunosuppression caused by some T lymphocyte subsets and its clinical application value in the evaluation，prognosis and immune intervention of sepsis.

Objective To investigate the effects of galectin-3(Gal3)on skin abscess development and activation of mast cells(MC)in mice infected with methicillin-resistant Staphylococcus aureus(MRSA).Methods Wild type mice and Gal3-knockout(Gal3-/-)mice,at 6～8 weeks of age,were divided into four groups:Wild type mice+PBS group,Wild type mice+MRSA group,Gal3-/-mice+PBS group,Gal3-/-mice+MRSA group,were subcutaneously injected with MRSA or the same volume of phosphate buffer saline,with five mice per group.The development and pathological changes of skin abscess were monitored and recorded.The bacterial load in skin tissues was compared,and the expression of associated cytokines,degranulation of MC,and the distribution of MC activation marker 5-hydroxytryptamine(5-HT)were detected.Results The skin of Wild type mice showed progressive abscesses after subcutaneous infection with MRSA,but the Gal3-/-mice showed smaller abscess areas.Compared to the Wild type mice+MRSA group,the Gal3-/-mice+MRSA group showed lower bacterial loading in the skin tissues(P＜0.01)and fewer infiltrating inflammatory cells with histopathological observation.The expression of cytokines,including IL-1β,TNF-α,IL-33,TGF-β,and IL-10,were significantly lower in Gal3-/-mice than Wild type mice(P＜0.05).The toluidine blue staining showed a large number of degranulated MCs in the skin tissues of the wild type mice+MRSA group,whereas only a few degranulated MCs were observed in the Gal3-/-mice+MRSA group.It was further found that the expression of 5-HT in Gal3-/-mice+MRSA group was significantly lower than that in wild-type mice+MRSA group with immunohistochemical staining.Conclusion Gal3 deficiency reduced the activation and degranulation of mouse skin MC after MRSA infection,resulting in changes to inflammatory responses and alleviating the severity of skin tissue abscesses.