Objective: To analyze the cause of discrepancy between ABO genotype B102/O01 and serological results in one case by PCR-SSP, to clarify the serological characteristics of this special blood group, and to explore relevant blood transfusion strategies. Methods: Blood group serological tests were performed on blood donor in August and December 2024, including forward and reverse ABO typing using tube method, H antigen identification, direct anti-human globulin test by tube method, red blood cell absorption-elution test, and determination of ABH blood group substance in saliva. Exons 1-7 of the ABO gene were amplified by PCR-SSP and sequenced. Results: The two separate serological tests consistently identified the donor as having an A B phenotype, but the results of gene sequencing indicated a B102/O01 genotype, showing an discrepancy between serological and genetic results. Conclusion: It is very likely that the blood type of the blood donor is B102/O01 with a microchimerism of type A, or an AB type masked by A type reference gene.

Hepatocellular carcinoma （HCC） is a malignant tumor with relatively high incidence and mortality rates worldwide， and its therapeutic resistance and recurrence mechanism are closely associated with liver cancer stem cells （LCSC）. This article systematically introduces the biological characteristics of LCSC and their key role in the progression of HCC， reviews the functional characteristics of the specific surface markers （such as EpCAM and CD133） and related signaling pathways （such as Wnt/β-catenin， TGF-β， and STAT3）， elaborates on the interaction between LCSC and tumor microenvironment， and summarizes the latest clinical treatment strategies targeting LCSC and the countermeasure for existing resistance mechanisms. The article points out that LCSC promote tumor development and progression through metabolic reprogramming and immune microenvironment remodeling， and it is proposed to establish a standardized detection system for LCSC specific markers and promote a triple synergistic therapeutic paradigm combining targeted therapy， immune regulation， and traditional chemotherapy， in order to provide new ideas for the clinical intervention of HCC.

Liver being substantial Yin and functional Yang maintain normal function of Qi， blood and meridians. In clinical practice， it is often found that pan-vascular lesions with atherosclerosis as the predominant pathological change often co-occur with metabolic dysfunction-associated fatty liver disease（MAFLD）. MAFLD leads to increased risk and worse prognosis for many pan-vascular diseases， including cardiovascular disease. Dysregulation of energy homeostasis disrupts the hepatic homeostasis of body use， and representative drugs to improve metabolism， such as metformin， sodium-glucose co-transporter 2 inhibitors， and glucagon-like peptide-1 agonists， not only have a clear cardiovascular benefit， potential improvement of MAFLD has also been demonstrated. The liver stores blood and the heart pumps blood， and liver diseases affect the heart， that's why the unsmoothness of vessels appears. So the treatment should from the standpoint of liver， restoring liver function， soothing the liver and nourishing heart， activating blood and dredging meridian. It is of great significance to explore in depth the pathogenesis and treatment of pan-vascular lesions caused by MAFLD， and to restore the energy homeostasis by adjusting the balance of liver Yin and Yang.

Liver being substantial Yin and functional Yang maintain normal function of Qi， blood and meridians. In clinical practice， it is often found that pan-vascular lesions with atherosclerosis as the predominant pathological change often co-occur with metabolic dysfunction-associated fatty liver disease（MAFLD）. MAFLD leads to increased risk and worse prognosis for many pan-vascular diseases， including cardiovascular disease. Dysregulation of energy homeostasis disrupts the hepatic homeostasis of body use， and representative drugs to improve metabolism， such as metformin， sodium-glucose co-transporter 2 inhibitors， and glucagon-like peptide-1 agonists， not only have a clear cardiovascular benefit， potential improvement of MAFLD has also been demonstrated. The liver stores blood and the heart pumps blood， and liver diseases affect the heart， that's why the unsmoothness of vessels appears. So the treatment should from the standpoint of liver， restoring liver function， soothing the liver and nourishing heart， activating blood and dredging meridian. It is of great significance to explore in depth the pathogenesis and treatment of pan-vascular lesions caused by MAFLD， and to restore the energy homeostasis by adjusting the balance of liver Yin and Yang.

Objective To investigate the relationship between serum ATP binding cassette subfamily A member 1(ABCA1)and fatty acid binding protein 4(FABP4)levels and insulin resistance(IR)and pregnan-cy outcome in patients with gestational diabetes mellitus(GDM).Methods A total of 121 patients with GDM admitted to the hospital from October 2019 to October 2023(GDM group)and 65 healthy pregnant women during the same period(control group)were selected,and the patients with GDM were divided into a poor group(50 cases)and a good group(71 cases)according to the pregnancy outcome.Serum ABCA1 and FABP4 levels were detected by enzyme-linked immunosorbent assay,and Pearson correlation coefficients were used to analyze the correlation between the two and the IR index(MOMA-IR)assessed by steady state model.With pregnancy outcome in GDM patients as the dependent variable,multivariate Logistic regression was used to determine its influencing factors,and receiver operating characteristic curve was used to evaluate the predictive efficacy of serum ABCA1 and FABP4 levels.Results Serum ABCA1 level in the GDM group was lower than those in the control group,and FABP4 level and HOMA-IR were higher than those in the control group(t=12.818,P＜0.001,t=17.219,P＜0.001,t=17.543,P＜0.001).In the GDM patients,HOMA-IR was nega-tively correlated with serum ABCA1 levels and positively correlated with serum FABP4 levels(r=-0.739,t=0.724,both P＜0.001).The independent risk factors for adverse pregnancy outcomes in GDM patients were HOMA-IR(OR=1.449,95％CI:1.161-1.810)and FABP4(OR=1.024,95％CI:1.011-1.037),and the independent protective factor was ABCA1(OR=0.302,95％CI:0.163-0.559).The area under the curve of serum ABCA1 combined with FABP4 level to predict pregnancy outcome in GDM patients was 0.877(95％CI:0.805-0.930),which was greater than that of serum ABCA1 and FABP4 levels alone,which were predic-ted by 0.786(95％CI:0.702-0.855),0.787(95％CI:0.703-0.856).Conclusion Reduced serum ABCA1 levels and elevated FABP4 levels are associated with IR and adverse pregnancy outcomes in patients with GDM,and the combination of the two has high predictive efficacy.

Cardiometabolic disease is a clinical syndrome with a causal relationship between metabolic abnormalities and cardiovascular damage. With its global incidence and related mortality rates continually rising， it has become a major health concern worldwide. The role of the gut microbiome and its metabolic products in cardiovascular metabolic health has received widespread attention， with gut microbiota dysbiosis considered a key factor in promoting the development of cardiometabolic disease. Dysbiosis disrupts the balance of the "heart-spleen-intestine" axis， leading to dysfunction of the spleen and intestines， which triggers metabolic disorders and accelerates the progression of cardiometabolic disease. Cardiac dysfunction can also negatively affect spleen and intestinal function， leading to imbalances in the heart and spleen， disharmony in Qi and blood， and exacerbating metabolic anomalies and further dysbiosis， thus forming a vicious cycle. From a modern biological perspective， the gut microbiome and its metabolic products can influence disease progression by modulating inflammatory responses and immune imbalances， leading to endothelial dysfunction and metabolic disorders， thereby increasing the risk of cardiometabolic disease. Additionally， the article proposes strategies for managing cardiometabolic disease by regulating the gut microbiome through a combination of Chinese and western medicine approaches. Traditional Chinese medicine（TCM） treatment starts from the gut microbiome， using the "heart-spleen-intestine" axis as a mediator to regulate cardiovascular metabolic health， highlighting the unique advantages of TCM in targeting the gut microbiome to treat cardiometabolic disease. This article takes the TCM theory of the "heart-spleen-intestine" axis as a starting point， discusses the pivotal role played by this axis in the connection between gut microbiome dysbiosis and the development of cardiometabolic disease， aiming to provide a new perspective for the integrated traditional Chinese and western medical research on cardiometabolic disease， offering scientific evidence and practical guidance to improve the prognosis and quality of life for patients.

Objective:To explore the effect and correlation of long non-coding RNA (lncRNA) IDI2-AS1/microRNA-33b-5p (miR-33b-5p)/nuclear receptor-associated protein NR4A2 competitive endogenous RNA (ceRNA) regulatory network on acute myocardial infarction (AMI), and to verify whether IDI2-AS1 regulates NR4A2 through miR-33b-5p to affect the occurrence and development of myocardial infarction.Methods:The miRNA and mRNA expression chips related to myocardial infarction were obtained from gene expression omnibus (GEO), and the differential expression was analyzed. The upstream regulatory mechanism of NR4A2 was predicted using TargetScan database. Thirty-two male C57/BL6 mice were divided into Sham group, AMI model group, miR-33b-5p mimic group [miR-33b-5p mimic lentivirus (5×10 7 TU) was injected locally into the heart tissue during ligation] and miR-33b-5p inhibitor group [miR-33b-5p inhibitor lentivirus (5×10 7 TU) was injected locally into the heart tissue during ligation] according to random number table method, with 8 mice per group. Left ventricular end-diastolic diameter (LVEDD) and left ventricular end-systolic diameter (LVESD) were asseessed by echocardiography, left ventricular fractional shortening (LVFS) and left ventricular ejection fraction (LVEF) were calculated. After the last weighing, the anesthetized mice were sacrificed and the heart tissues were taken. Masson staining of the heart tissues was observed under light microscope, myocardial collagen volume fraction (CVF) and infarct size were calculated. Cardiomyocytes of SPF grade SD rats were collected. They were divided into normal control group (control group), ischemia-hypoxia model group, miR-33b-5p mimic transfection group (miR-33b-5p mimic transfection group before ischemia and hypoxia treatment) and miR-33b-5p inhibitor transfection group (miR-33b-5p inhibitor transfection group before ischemia and hypoxia treatment). The activity of caspase-3/7 in cardiomyocytes was measured. The levels of interleukins (IL-1β, IL-6) and tumor necrosis factor-α (TNF-α) were detected by enzyme-linked immunosorbent assay (ELISA). The levels of malondialdehyde (MDA), superoxide dismutase (SOD), creatine kinase (CK), MB isoenzyme of creatine kinase (CK-MB) and lactate dehydrogenase (LDH) were detected by colorimetry. Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of apoptosis-related proteins Bax and Bcl-2, cytochrome C (Cyt C) and IDI2-AS1/miR-33b-5p/NR4A2 regulatory axis genes. Results:The myocardial infarction microarray analysis showed that NR4A2 expression was significantly up-regulated in myocardial infarction, with predicted upstream regulatory mechanisms indicating its possible influence through the IDI2-AS1/miR-33b-5p/NR4A2 regulatory axis. Echocardiographic detection showed that compared with AMI model group and miR-33b-5p inhibitor group, LVEF and LVFS in the heart tissue of mice in miR-33b-5p mimic group were significantly increased, while the levels of LVEDD, LVESD, CK, CK-MB and LDH were significantly decreased, with statistical significance. Light microscope showed myocardial fibrosis and myocardial infarction in AMI model group and miR-33b-5p inhibitor group. In the miR-33b-5p mimic group, the degree of myocardial fibrosis was decreased and the myocardial infarction size was significantly reduced. Compared with AMI model group and miR-33b-5p inhibitor group, the levels of MDA, IL-1β, IL-6, TNF-α and the expressions of Bax and Cyt C in the heart tissue of mice in miR-33b-5p mimic group were significantly decreased, while the levels of SOD and Bcl-2 expression were significantly increased, and the differences were statistically significant. The expressions of IDI2-AS1 and NR4A2 in the heart tissue of mice in miR-33b-5p mimic group were significantly lower than those in AMI model group and miR-33b-5p inhibitor group [IDI2-AS1 (2 -ΔΔCt): 1.96±0.08 vs. 2.73±0.08, 3.10±0.05, NR4A2 (2 -ΔΔCt): 2.36±0.07 vs. 3.16±0.08, 3.80±0.08, all P < 0.01]. The expression of miR-33b-5p was significantly higher than that of AMI model group and miR-33b-5p inhibitor group (2 -ΔΔCt: 0.88±0.07 vs. 0.57±0.07, 0.23±0.01, both P < 0.01). The cell experiment results showed that the caspase-3/7 activity of rat neonatal cardiomyocytes in the miR-33b-5p mimic transfection group was significantly lower than that in the ischemia-hypoxia model group and the miR-33b-5p inhibitor transfection group, suggesting that miR-33b-5p can significantly reduce the apoptosis level of the ischemia-hypoxia model. The levels of peroxidation and inflammation indexes, important genes of apoptosis pathway and the expression of IDI2-AS1/miR-33b-5p/NR4A2 regulatory axis of rat neonatal cardiomyocytes in all groups were consistent with the above. Conclusion:IDI2-AS1 can regulate NR4A2 through miR-33b-5p, thus affecting the occurrence and development of AMI.

Objective To explore the relationship between optic nerve tissue thickness and enlarged perivascular space(EPVS)in patients with recent small subcortical infarct(RSSI).Methods A total of 72 RSSI patients admitted to Department of Neurology of Changzhou Second People's Hospital affiliated to Nanjing Medical University from November 2021 to December 2022 were re-cruited in this study.All patients underwent cranial MRI,optical coherence tomography(OCT)and fundus photography.The thickness of retinal nerve fiber layer(RNFL)and ganglion cell-inner plexiform layer(GCL-IPL)were measured by software semi-automatic segmentation,and fundus lesions such as fundus hemorrhage and exudation were observed through fundus photography.The severity of EPVS was graded by visual quantitative evaluation,and then the patients were divided into no to mild EPVS group(32 cases)and moderate to severe EPVS group(40 cases).Basal gan-glia EPVS(BG-EPVS)and semioval center EPVS(CSO-EPVS)were further graded and grouped into no to mild BG-EPVS group(31 cases)and moderate and severe BG-EPVS group(41 cases),and no to mild CSO-EPVS group(39 cases)and moderate and severe CSO-EPVS group(33 cases).The clinical data of each group were compared separately.Results There were statistical differences in age,ratio of hypertension and thicknesses of RNFL and GCL-IPL between the no to mild EPVS group and the moderate to severe EPVS group(P＜0.05,P＜0.01),between the no to mild BG-EPVS and the moderate to severe BG-EPVS group(P＜0.01).Multivariate logistic re-gression analysis showed that age(OR=1.207,95％CI:1.028-1.416,P=0.021),hypertension(OR=7.264,95％CI:1.079-11.929,P=0.042),RNFL thickness(OR=0.753,95％CI:0.617-0.915,P=0.004)and GCL-IPL thickness(OR=0.848,95％CI:0.758-0.949,P=0.004)were independent factors of BG-EPVS grade.Spearman correlation analysis indicated that the thicknes-ses of RNFL and GCL-IPL were negatively correlated with BG-EPVS grade(r=-0.571,r=-0.649,P＜0.01).Conclusion RNFL and GCL-IPL thicknesses are independent factors for BG-EPVS grade and negatively correlated with BG-EPVS grade.OCT has important clinical value in assessing the severity of EPVS lesions in RSSI patients.

Objective:To study the expression of zonula occludens-1(ZO-1) in neonates with necrotizing enterocolitis (NEC) and to explore the effects of disialyllacto-N-tetraose (DSLNT), a compound extracted from human milk, on the intestinal barriers in rat model of NEC.Methods:(1) Human study: From Feb 2013 to Dec 2020, the pathological samples of ileum tissue from 21 neonates (12 patients with NEC and 9 with intestinal atresia) from Pathology Department of our hospital were collected. The expressions of ZO-1 in these samples were examined using immunohistochemistry (IHC) method. (2) Animal study: A total of 28 newborn rats were randomly assigned into control group ( n=8), NEC group ( n=10) and DSLNT+NEC group ( n=10). Experimental NEC model was established based on hypoxia (95%N 2 10 min) /cold exposure (4 ℃ 10 min) three times a day for consecutive 3 days. DSLNT+NEC group were fed with formula+DLSNT (300 μmol/L) during hypoxia/cold exposure. All the surviving rats were sacrificed at the end of the experiment (72 h) and the terminal ileum tissues were collected. Hematoxylin-Eosin (HE) staining was used to evaluate tissue damage and Western blotting was used to determine the expressions of ZO-1. (3) Cellular study: Bacterial lipopolysaccharide (LPS) was used to establish a cellular inflammation model in human intestinal epithelial cell lines (Caco-2) and DSLNT (300 μmol/L) was applied to this model. Thiazolyl blue assay was used to examine cell viabilities and immunofluorescence assay was used to detect ZO-1 expression. The effects of DSLNT on cell growth and tight junctions of Caco-2 cells were analyzed. Results:(1)Human study: The villi of mucous layer of the lesion were damaged in NEC patients. ZO-1 expressions at the epithelial junction of NEC patients were decreased compared with intestinal atresia patients and non-lesion intestines of NEC patients. (2)Animal study: Apical extrusion, necrosis and shedding of epithelial cell were seen at the lesions in NEC group. The expression of ZO-1 in NEC group was significantly lower than the control group and DSNLT+NEC group ( P<0.05).DSNLT+NEC group had higher survival rates (8/10 vs. 6/10) and lower ileum inflammatory pathological scores [2.0(1.0, 2.8) vs. 3.5(3.0, 4.0)] than NEC group. (3) Cellular study: Caco-2 cells exposed to LPS showed inhibited cell growth and decreased ZO-1 immunofluorescence staining. Caco-2 cells in the DSLNT+LPS group showed better viability than LPS group and were comparable with the control group. The expression of ZO-1 was significantly increased in the DSLNT+LPS group. Conclusions:Tight junction injury of the intestinal epithelial cell is an important characteristic of NEC. ZO-1 is a potential target for the prevention and treatment of NEC. DSLNT may protect the neonatal intestines by modulating the expression of ZO-1 and keeping tight junction integrity.

Objective:To explore the current situation and related factors of asphyxia prevention and management for the elderly in elderly care institutions, so as to provide a reference for improving the safety management of elderly care institutions.Methods:This research was a cross-sectional survey. From December 2020 to January 2021, convenience sampling was used to select 43 elderly care institutions from the China Aged Care Alliance. The self-designed questionnaire was used to collect the content related to asphyxia prevention and management for the elderly in institutions, and analyze its current situation and related factors. A total of 43 questionnaires were distributed in this survey, and 43 were recovered, with a recovery rate of 100%.Results:Among the 43 elderly care institutions, and 3 (6.98%) were state-run, and 22 (51.16%) were private, and 18 (41.86%) were public and private. The average occupancy rate was (67.63±20.59) %, and the average proportion of moderate to severe disability among the elderly residents was (80.10±20.25) %. The proportions of elderly care institutions that formulated systems and regulations for monitoring and early warning of asphyxia events, reporting of asphyxia events, analysis and improvement of asphyxia incidents in the elderly were 53.49%, 51.16%, and 51.16%, respectively. The proportions of elderly care institutions that carried out asphyxia risk screening, swallowing function assessment, and swallowing function training for the elderly residents were 65.12%, 51.16%, and 46.51%, respectively.Conclusions:Most elderly care institutions have problems in the prevention and management of asphyxia for the elderly, which needs to be improved. It is recommended that building a complete asphyxia prevention and management system, carrying out risk screening, assessment and training of swallowing function for high-risk groups, strengthening the professional training of agency leaders and caregivers, and seeking continuous external professional support so as to improve the safety management system of elderly care institutions, and ensure the quality of life and personal safety of the elderly.