[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.

Micro/nanoplastics (MNPs), recognized as emerging environmental pollutants, are widely distributed in natural environments. Due to their small particle size and significant migratory capacity, MNPs can infiltrate diverse environmental matrices, then invade and accumulate in the organism via the skin, respiration, and digestion. Recently, concerns have grown over the detrimental effects and potential toxicity of MNPs on reproductive health. This review summarized published epidemiological and toxicological studies related to MNPs exposure and their effects on reproductive health. Firstly, this review critically examined the current landscape of epidemiological evidence and found that MNPs (e.g., polystyrene, polypropylene, polyvinyl chloride, polyethylene, etc.) are present in various biological specimens from both males and females, and their presence may be associated with an increased risk of reproductive disorders. Secondly, extensive toxicological studies revealed that MNPs exposure induces reproductive health damage through mechanisms such as disrupting the microstructure of reproductive organs and altering molecular-level expressions. Oxidative stress, inflammatory responses, and apoptosis are identified as potential links between MNPs exposure and reproductive damage. Finally, this review addressed the prevalent shortcomings in existing studies and proposed future directions to tackle the challenges posed by MNPs-induced reproductive harm. These insights aim to inform strategies for safeguarding public reproductive health and ecological security, providing a scientific foundation for mitigating risks associated with MNPs pollution.

Ketone body metabolism plays a significant role in the development and progression of diabetic retinopathy(DR), which closely related to the system and local metabolic disorders as a major microvascular complication of diabetes mellitus. Previous research has established a close relationship between dyslipidemia and DR progression. Ketone bodies, comprising β-hydroxybutyrate, acetoacetate, and acetone, are metabolic products generated from fat breakdown when glucose metabolism is impaired. Studies have revealed that ketone body metabolism is intricately linked to multiple pathophysiological processes in DR, including oxidative stress, inflammatory responses, and neurodegeneration within retinal cells. This article provides a review exploring the impact of ketone body metabolism on the pathogenesis of DR, and systematically reviews the latest research progress on the impact of ketone bodies on the core pathological links such as retinal vascular barrier destruction, glial cell activation and angiogenesis through metabolic reprogramming, epigenetic modification and cell signal transduction, so as to provide a theoretical basis for in-depth understanding of the metabolic driving mechanism of DR.

Objective To discover the host factor interacting with severe acute respiratory syndrome coronavirus-2(SARS-CoV-2)papain-like protease(PLpro)and explore the potential mechanism.Methods The second-generation proximity-dependent biotin identification(BioID2)approach combined with mass spectrometry analysis was used to search for the potential host factors.Immunofluorescence and co-immunoprecipitation(Co-IP)assay were used to verify the interactions between DEAD-box helicase 3(DDX3)and PLpro.The influence of PLpro on DDX3-inhibitor of kappa B kinase ε(IKKε)-TANK-binding kinase 1(TBK1)and DDX3-mitochondrial antiviral signaling protein(MAVS)complexes was also investigated by Co-IP.The effect of PLpro on interferon-β(IFN-β)immune pathway and the protease activity on substrates were studied via luciferase activity assay.Results DDX3 could co-locate and interact with PLpro intracellularly.PLpro might possibly inhibit both the formation of DDX3-MAVS complex and the interactions between DDX3-IKK-ε-TBK1.PLpro could negatively regulate type Ⅰ interferon pathway.Overexpression of DDX3 could lead to a significant increase in the cleavage activity of PLpro/PLP-TM that might be significantly decreased in case of inventions with DDX3 expressions.Conclusion DDX3 may be one of the host factors that interact with SARS-CoV-2 PLpro.PLpro negatively regulates IFN-β immune pathway induced by DDX3,which may provide a favorable immune environment for virus replication.

Objective:To identify the key genes in the process inhibiting inflammation by overexpression adenovirus-mediated pigment epithelium-derived factor ( PEDF) gene in human monocytic leukemia cells THP1. Methods:Proteomic analysis of THP1 overexpressing adenovirus-mediated PEDF gene was performed.The THP1 cells were divided into GFP and PEDF groups, transfected with GFP and PEDF adenovirus, respectively.The THP1 cells were divided into mannitol group, high glucose group, high glucose+ GFP group, and high glucose+ PEDF group, which were cultured with mannitol for 4 days, anhydrous glucose for 4 days, GFP adenovirus for 3 days, and PEDF adenovirus for 3 days, respectively.The Pedf-/- mice were divided into Pedf-/- group and Pedf-/- diabetes group according to the random table method, with 12 mice in each group.Another 10 C57BL/6 mice were taken as the control group.Mouse retinas were collected for experiments.The mRNA expression levels of differentially expressed genes (DEGs) in retina and THP1 cells were verified by real-time fluorescence quantitative PCR.The DEGs were intersected with the GSE5504 dataset, and the protein-protein interaction (PPI) network was built using the String database.Modules of the PPI were extracted using the Cytoscape software and the MCODE application.Intersections were taken with the Set1 dataset and key genes were found.The expression levels of key genes in THP1 cells and Pedf-/- mice were verified by Western blot.The feeding and operation of experimental animals were in accordance with the regulations of the State Science and Technology Commission on the management of experimental animals and approved by the Animal Management and Use Committee of Tianjin Medical University (No.TTYY2023120217). Results:Through proteomics and bioinformatics analysis, 105 DEGs in the Set1 dataset were screened.The results of real-time PCR showed that the relative expression levels of ARF5, TCF25 and KCTD9 mRNA were significantly higher and the relative expression levels of RNPS1, CSF1R, OGA, IBA57 and MGST2 mRNA were significantly lower in PEDF group than in GFP group, showing statistically significant differences (all at P<0.001).There were significant overall differences in the relative expression levels of down-regulated TCF25, KCTD9 and ARF5 mRNA and up-regulated CSF1R, RNPS1 and IBA57 mRNA among control group, Pedf-/- group and Pedf-/- diabetes group ( F=64.057, 27.561, 37.179, 65.757, 44.024, 34.248; all at P<0.001).Compared with control group, the relative expression levels of TCF25, KCTD9 and ARF5 mRNA were decreased and the relative expression levels of CSF1R and RNPS1 mRNA were increased in Pedf-/- group, showing statistically significant differences (all at P<0.05).Compared with control group, the relative expression levels of TCF25, KCTD9 and ARF5 mRNA were decreased and the relative expression levels of CSF1R, RNPS1 and IBA57 mRNA were increased in Pedf-/- diabetes group, showing statistically significant differences (all at P<0.05).Compared with Pedf-/- group, the relative expression level of TCF25 mRNA was decreased and the relative expression levels of CSF1R, RNPS1 and IBA57 mRNA were increased in Pedf-/- diabetes group, showing statistically significant differences (all at P<0.05).After intersection with the GSE5504 dataset, 20 differential proteins were obtained, which were mainly enriched in positive regulation of gene expression, positive regulation of ERK1 and ERK2 cascade, positive regulation of insulin secretion involved in cell response to glucose stimulation and antigen processing and presentation pathways.The key gene CSF1R was screened by constructing PPI network and MCODE plugin in Cytoscape software.Western blot results showed that the expression levels of CSF1R in high glucose group and high glucose+ GFP group were 1.961±0.085 and 1.000±0.069, which were higher than 1.000±0.072 in mannitol group and 0.469±0.079 in high glucose+ PEDF group, respectively, and the differences were statistically significant ( t=14.940, 8.765; both at P < 0.01).The expression of CSF1R in the retina of Pedf-/- diabetes group was 1.633±0.192, which was higher than 1.000±0.050 in Pedf-/- group, and the difference was statistically significant ( t=5.537, P<0.01). Conclusions:CSF1R may be a key gene and therapeutic target for the inhibition of inflammation by overexpression of adenovirus-mediated PEDF gene in THP1 cell.

Arsenic is a widely occurring metalloid element in the natural environment and is one of the primary carcinogens identified by the World Health Organization (WHO), but the specific carcinogenic mechanism is currently unclear. In recent years, through toxicological studies on arsenic, it has been found that exposure to arsenic can affect cellular metabolism in the body, which may be closely related to the carcinogenic mechanism of arsenic. Therefore, the authors review the research progress on arsenic exposure-induced effects on glucose metabolism, lipid metabolism, and amino acid metabolism, with a view to providing a theoretical basis for the study of the mechanism of arsenic carcinogenesis.

Objective To explore the relationship between the expression levels of serum cingulate protein(CGN)and polyligand glycan 1(SDC-1)and the disease condition and outcome of hypertensive basal ganglia hemorrhage(HBGH).Methods A total of 123 patients with HBGH admitted to the Second People's Hospi-tal of Lianyungang from February 2019 to February 2022 were selected as the study objects,and 120 healthy volunteers who underwent physical examination in the hospital during the same period were selected as the health group.Serum CGN and SDC-1 expression levels were detected in the two groups.According to the dis-ease outcome,the patients were divided into the improved group(92 cases)and the deteriorated group(31 ca-ses).Receiver operating characteristic(ROC)curve and the area under the curve(AUC)were used to analyze the predictive value of serum CGN and SDC-1 expression levels on the disease outcome of patients with HB-GH.Results Serum CGN and SDC-1 expression levels in the severe group were higher than those in the mod-erate group and the mild group,and serum CGN and SDC-1 levels in the moderate group were higher than those in the mild group,and the differences were statistically significant(P＜0.05).Serum CGN and SDC-1 expression levels in HBGH patients in three groups were higher than those in health group,and the differences were statistically significant(P＜0.05).Serum CGN and SDC-1 expression levels in the deteriorated group were higher than those in the improved group,and the differences were statistically significant(P＜0.05).The AUC of serum CGN and SDC-1 for predicting the disease outcome of HBGH patients was 0.742(95％CI:0.792-0.697)and 0.861(95％CI:0.906-0.910),respectively,and the AUC of the combination of the two was 0.917(95％CI:0.962-0.870).The amount of blood loss and ventricular rupture in the deteriorated group were higher than those in the improved group,and the Glasgow Coma Scale(GCS)score on admission was lower than that in the improved group,and the differences were statistically significant(P＜0.05).Multi-variate Logistic regression analysis showed that serum CGN≥51.63 pg/mL(OR=3.815),serum SDC-1≥450.67 μg/L(OR=4.230)and GCS score ≤8(OR=5.333)were the influencing factors for disease outcome of HBGH patients(P＜0.05).Conclusion The increased expression levels of serum CGN and SDC-1 are closely related to the disease aggravation and the deterioration of the disease outcome in patients with HBGH,and they have certain predictive value for the disease outcome in patients with HBGH.

TCM believes that spleen deficiency is the root cause of emotional abnormalities in chronic fatigue syndrome (CFS), and clinical treatment often involves the heart, liver and kidney. TCM therapy has a significant efficacy in CFS emotional abnormalities. It is mostly treated with oral administration of TCM, acupuncture, moxibustion and massage therapy. It may play a therapeutic role by improving oxidative stress and immune inflammation, regulating nerve-endocrine, controlling energy metabolism and other ways. It is suggested to establish the syndrome differentiation standard of CFS emotional abnormality in the future, so as to improve the accuracy of syndrome differentiation and treatment; form a perfect treatment guide or expert consensus to guide the standardized application of various internal and external treatment methods; explore objective indicators based on the pathogenesis, and focus on the morphological and functional changes of disease target brain regions with the help of neuroimaging techniques, so as to improve the diagnosis and prognosis evaluation of CFS; based on the guidance of TCM theory, improve the CFS emotional abnormal animal modeling method.

TCM functional exercises are the important means of TCM to prevent and cure diseases. By adjusting the bones and muscles externally, adjusting the heart and organs internally, promoting blood circulation, improving sleep disorders, enhancing metabolism and immune capacity, the aim of preventing and treating diseases, prolonging life span, and strengthening the body is achieved. TCM exercises have a significant effect on the treatment of various types of fatigue such as chronic fatigue syndrome, Exercise-induced fatigue, post-stroke fatigue, and cancer-related fatigue.