Objective To investigate the relationship between the expression of serum angiopoietin-like protein 8(ANGPTL8)and Kruppel-like factor 2(KLF2)and the degree of coronary artery disease and the occurrence of major adverse cardiac events(MACE)in patients with acute myocardial infarction(AMI).Methods A total of 106 patients with AMI who were hospitalized in our hospital from January 2021 to June 2023 were selected as the research objects.According to the degree of coronary artery disease,the patients were grouped into mild group(52 cases)and severe group(54 cases).According to the occurrence of MACE,the patients were grouped into MACE group(18 cases)and non-MACE group(88 cases).General patient data were collected.Serum ANGPTL8 and KLF2 levels were detected by enzyme-linked immunosorbent assay(ELISA).Spearman correlation analysis was applied to analyze the correlation between serum ANGPTL8 and KLF2 levels and Gensini score in AMI patients.Multivariate logistic regression was applied to analyze the influencing factors of coronary artery disease degree in patients with AMI.Receiver operating characteristic(ROC)curves were drawn to analyze the value of serum ANGPTL8 and KLF2 levels in predicting the occurrence of MACE in AMI patients.Results The proportions of patients with history of hypertension and hyperlipidemia,systolic blood pressure,diastolic blood pressure,levels of triacylglycerol(TG),N-terminal pro-B-type natriuretic peptide(NT-proBNP),cardiac troponin I(cTnI),Gensini score,and level of serum ANGPTL8 in the severe group were higher than those in the mild group(P<0.05).The level of high-density lipoprotein cholesterol(HDL-C)and serum KLF2 in the severe group were lower than those in the mild group(P<0.05).The number of lesions in the mild group and the severe group was statistically obvious(P<0.05).Serum ANGPTL8 level in AMI patients was positively correlated with Gensini score(r=0.638,P<0.05),and serum KLF2 level was negatively correlated with Gensini score(r=-0.612,P<0.05).History of hypertension,hyperlipidemia,cTnI and ANGPTL8 were risk factors for the progression of severe coronary artery disease in patients with AMI(P<0.05),while HDL-C and KLF2 were protective factors(P<0.05).The serum level of ANGPTL8 in AMI patients in the MACE group was higher than that in the non-MACE group(P<0.05),and the serum KLF2 level in the MACE group was lower than that in the non-MACE group(P<0.05).The area under the curve of serum ANG-PTL8 and KLF2 levels and their combination in predicting the occurrence of MACE in AMI patients was 0.740(95%CI:0.646～0.820),0.799(95%CI:0.710～0.870),and 0.806(95%CI:0.717～0.876),respectively.Conclusion The expressions of serum ANGPTL8 and KLF2 are closely related to the degree of coronary artery disease in patients with AMI,and have certain predictive value for the occurrence of MACE.

Aim To explore the key genes in the blood transcriptome of atherosclerosis patients based on blood transcriptomics.Methods Three datasets GSE12288,GSE27034 and GSE90074 were extracted from the GEO data-base and performed the merging and normalization processing.The differential genes in peripheral blood samples of ather-osclerosis patients and controls were analyzed,and enrichment analysis of differentially expressed genes were performed.Then weighted gene co-expression network analysis was performed for all genes.Using differentially expressed genes to construct protein-protein interaction(PPI)network,and using CytoHubba to screen key genes based on co-expression net-work and PPI network.And the expression levels of key genes were detected by RT-qPCR.Results 74 down-regu-lated genes and 145 up-regulated genes were identified between atherosclerosis patients and controls.GO and KEGG en-richment analyses revealed that they were significantly enriched in neutrophil activation,granulocyte activation,cytokine-cytokine receptor interaction and chemokine signaling pathway.In addition,the top 10 genes in the co-expression network and the top 20 genes in the PPI network were also identified,in which PRF1,NKG7,GZMB and CCL5 played a high core role in the PPI network and co-expression network.The RT-qPCR results showed that compared with the non coronary atherosclerosis controls,the mRNA expression levels of PRF1 and GZMB in peripheral venous blood peripheral blood mono-nuclear cell(PBMC)of coronary atherosclerosis patients were significantly reduced(P＜0.05),while the mRNA expres-sion levels of NKG7 and CCL5 were significantly increased(P＜0.05).Conclusion PRF1,GZMB,NKG7 and CCL5 may be key genes in the blood transcriptome of atherosclerosis patients,and are expected to be potential biomarkers for diagnosis and treatment of atherosclerosis.

A 2-year-old boy was admitted to Shanghai Children's Medical Center, Shanghai Jiao Tong University School of Medicine in Nov 30^th, 2018, due to polydipsia, polyphagia, polyuria accompanied with increased glucose levels for more than 2 weeks. He presented with symmetrical short stature [height 81 cm (－2.2 SD), weight 9.8 kg (－2.1 SD), body mass index 14.94 kg/m² (P₁₀-P₁₅)], and with no special facial or physical features. Laboratory results showed that the glycated hemoglobin A1c was 14%, the fasting C-peptide was 0.3 ng/mL, and the islet autoantibodies were all negative. Oral glucose tolerance test showed significant increases in both fasting and postprandial glucose, but partial islet functions remained (post-load C-peptide increased 1.43 times compared to baseline). A heterozygous variant c.1366C>T (p.R456C) was detected in GATA6 gene, thereby the boy was diagnosed with a specific type of diabetes mellitus. The boy had congenital heart disease and suffered from transient hyperosmolar hyperglycemia after a patent ductus arteriosus surgery at 11 months of age. Insulin replacement therapy was prescribed, but without regular follow-up thereafter. The latest follow-up was about 3.5 years after the diagnosis of diabetes when the child was 5 years and 11 months old, with the fasting blood glucose of 6.0-10.0 mmol/L, and the 2 h postprandial glucose of 17.0-20.0 mmol/L.
Male ; Child ; Humans ; Child, Preschool ; Infant ; Diabetes Mellitus, Type 2/complications* ; Mutation, Missense ; C-Peptide/genetics* ; China ; Insulin/genetics* ; Glucose ; Blood Glucose ; GATA6 Transcription Factor/genetics*

Human immunodeficiency virus-associated pulmonary arterial hypertension (HIV-PAH) is a long-term cardiovascular complication of AIDS patients, with an incidence of about 0.5%. The onset of HIV-PAH is insidious and lack of specific symptoms with poor prognosis. The pathogenesis is complicated while the bystander effect of HIV or the complication of HIV is possible mechanism. Echocardiography is an important diagnostic method and facilitates early screening of patients. At present, there is no specific drug targeted HIV-PAH, and the treatment strategy is to follow the treatment recommendations for idiopathic pulmonary arterial hypertension on the basis of highly active antiretroviral therapy, while the interaction between two types of drugs should be considered. This paper will mainly focus on the pathogenesis and treatment of HIV-PAH.

AIM: To investigate the relationship between serum soluble growth stimulation expressed gene 2 protein(sST2) level and coronary artery complex lesions and their severity. METHODS: A total of 430 patients, who were sequentially admitted to hospital for selective coronary artery angiography, were divided into control group (non-coronary heart disease group, 136 patients), simple lesions group of coronary heart disease (86 patients), complex lesions group (208 patients). To quantitative evaluate the complexity of coronary artery lesions, Syntax scores were further performed on patients in complex lesions groups, including 139 patients in the low-risk group, 36 patients in the medium-risk group, and 33 patients in the high-risk group. The serum soluble ST2 level of each group of patients was tested by means of ELISA. Spearman correlation analysis was used for the correlation between the level of soluble ST2 and the severity of coronary complex lesions. RESULTS: In 430 subjects, the soluble ST2 level of all patients with coronary heart disease (including simple lesions and complex lesions) was significantly higher than that of the control group [(3 449±1 250) vs. (2 743±961) pg/mL, P<0.001]; the sST2 levels of patients in the coronary artery simple lesions group, complex lesions low-risk group, medium-risk group and high-risk group were (3 200±1 406), (3 338±1 064), (3 728±1 228) and (4 261±1 235) pg/mL respectively, and the differences of sST2 levels among above four groups were statistically significant (P<0.001). Logistic regression analysis showed that sST2 was independently associated with coronary heart disease (OR=1.001, P<0.001) and sST2 was independently associated with the severity of coronary artery complex lesions (OR=1.001, P<0.001). Spearman-related analysis shows that the expression levels of sST2 are positively related to the severity of coronary artery lesions (rs: 0.543, P<0.001). The ROC curve showed that the area under the curve of sST2 for complex coronary lesions was AUC=0.726. CONCLUSION: Serum soluble ST2 level may be an important predictor of complicated coronary artery disease.

Objective To study whether Nεcarboxymethyl lysine(CML)can form a good molecular docking with the scavenger receptor CD36and induce a stable interaction.Methods The interaction between CML and CD36was studied by co-immunoprecipitation.The binding mode and affinity of CD36to CML were tested using AutoDock 4.2,iBabel and XQuartz-2.7.7software respectively. Results Co-immunoprecipitation showed that anti-CD36antibody magnetic bead could precipitate CD36from the total protein in RAW264.7cells and anti-CML could detect CD36 binding CML.CD36had a good molecular docking with CML,CD36and CML interacted stably with each other.The affinity of CML to 4Q4Bprotein structure of CD4extracellular domain was -29.62kJ/mol.ARG82,ASN71and THR70were the products of amino acid receptor interaction. Further docking analysis showed that CML could form 3interacting hydrogen bonds with 4Q4B,and the docking prediction inhibition constant was 6.92with a root mean square deviation of 2.54.Conclusion A good molecular docking between CML and 4Q4Bprotein structure of CD36extracellular domain can induce a stable interaction between CML and CD36.Hydrogen bonding is the main interaction mode.

Objective     To explore the surgical management method and outcome of functional univentricular heart with total anomalous pulmonary vein drainage (UVH-TAPVD). Methods     We reviewed the surgical procedures and results for 44 UVH-TAPVD patients in our hospital between the year 2010 and 2016. There were 34 males and 10 females. The age of the patients was 312 (77-4 220) d when they accepted the first surgical treatment. Results     There were 8 deaths in stage Ⅰ palliation, 1 death in stage Ⅱ palliation and 5 deaths during the follow-up. The overall survival rate was 68.2% (30/44). Glenn operation was undertaken in 34 patients with 5 deaths. Fontan operation was undertaken in 9 patients with 2 deaths. Conclusion     Surgical strategies for UVH-TAPVD should be planned according to different anatomical and pathophysiological conditions in different patients. Staged palliations can reduce mortality and morbidity. But pulmonary venous obstruction and heterotaxy syndrome are still risk factors for these patients.

Objective: To investigate whether CD137-CD137L signaling can affect the autophagy of mouse vascular smooth muscle cells(VSMCs) through JNK signal pathway. Methods: Primary culture of C57BL/6J mouse thoracic aorta VSMCs was performed by tissue block adherence method. VSMCs between the third to fifth passages were isolated and cultured. VSMCs were divided into 4 groups: control group, CD137 agonist group, JNK inhibition group, and DMSO group. VSMCs in CD137 agonist group were treated with recombinant protein of CD137L (10 μg/ml), VSMCs in JNK inhibition group were treated with JNK inhibitor SP600125 (10 μmol/L) for 30 minutes followed by recombinant protein of CD137L (10 μg/ml) and DMSO group was treated with the same amount of DMSO in JNK inhibition group for 30 minutes, then added recombinant protein of CD137L (10 μg/ml). Western blot was used to detect the protein expression of p-JNK, LCⅡ and p62 in each group. Fluorescence microscopy was used to track the changes of autophagy in cells which was infected with adenovirus expressing tandem mRFP-GFP-LC3. Transmission electron microscope (TEM) was used to observe intracellular autophagosomes and autolysosomes. Results: (1) Compared with the control group, stimulating CD137-CD137L axis by recombinant protein of CD137L significantly upregulated the expression of p-JNK, LCⅡ and p62 (1.15±0.19 vs. 0.72±0.21, P<0.05;1.03±0.13 vs. 0.59±0.15, P<0.05, and 1.10±0.19 vs. 0.76±0.15, P<0.05). These effects could be reduced by JNK inhibitor (0.61±0.21 vs. 1.15±0.19, P<0.05;0.74±0.11 vs. 1.03±0.13, P<0.05, and 0.21±0.12 vs. 1.10±0.19, P<0.05). The expression of these proteins in DMSO group remained unchanged compared with CD137 agonist group (P>0.05). (2) Changes of autophagy in cells of various group: the number of total fluorescent spots and yellow fluorescent spots in CD137 agonist group was significantly increased compared to control group (total fluorescent spots:(93.00±14.11)/cell vs. (52.33±9.61)/cell, P<0.05, and (64.33±6.81)/cell vs. (25.67±3.51)/cell, P<0.05), moreover, the number of yellow fluorescent spots was higher than the red fluorescent spots fluorescent spots in CD137 agonist group. Compared with CD137 agonist group, pretreatment with JNK inhibitor significantly reduced the number of total fluorescent spots and yellow fluorescent spots ((53.00±3.17)/cell vs. (93.00±14.11)/cell, P<0.05,and (15.33±4.51)/cell vs. (64.33±6.81)/cell, P<0.05). The red fluorescent spots were higher than the yellow fluorescent spots in JNK inhibition group. The number of total fluorescent spots and yellow fluorescent spots in CD137 agonist group was not affected by pretreatment with DMSO (P>0.05). (3) The number of intracellular autophagosomes and autolysosomes was significantly higher in CD137 agonist group than in control group((17.67±6.03)/cell vs. (5.67±2.52)/cell, P<0.05), and the number of autophagosomes was higher than that of autolysosomes in CD137 agonist group((14.00±4.00)/cell vs. (3.67±2.08)/cell, P<0.05). The number of intracellular autophagosomes and autolysosomes was significantly lower in JNK inhibition group compared to CD137 agonist group((5.67±4.04)/cell vs. (17.67±6.03)/cell, P<0.05) and the number of autophagosomes was lower than that of autolysosomes in JNK inhibition group((1.33±1.53)/cell vs. (4.33±2.52)/cell, P<0.05). The number of intracellular autophagosomes and autolysosomes was similar between DMSO group and CD137 agonist group (P>0.05). Conclusion CD137-CD137L signal may influence autophagy of mouse VSMCs via JNK pathway.

Objective: To explore whether CD137-CD137L interaction could induce mouse vascular smooth muscle cells(VSMCs) calcification via P38 MAPK signaling. Methods: (1) Mouse VSMCs obtained from 8-week old male C57 mice were cultured by using method of tissue piece inoculation.The cells from 3 to 8 passage were divided into 4 groups: control group, agonist-CD137 group(recombinant CD137L protein), anti-P38 group(agonist-CD137 group+P38 inhibitor), single anti-P38 group(P38 inhibitor). The calcification was induced by adding a mixture of 10 mmol/L β-glycerophosphate+10^-8 mol/L dexamethasone+10^-7 mol/L insulin in the culture medium.Immunofluorescence was used to observe the changes of VSMCs markers(α-SMA and OPN).Real time-PCR was used to observe the mRNA expression of OPN and RUNX-2. Western blot was used to observe the protein expression of p-P38, OPN and RUNX-2. The level of cell calcification was observed by detecting alkaline phosphatase activity and calcium concentration. (2) The degeree of local calcium deposition was also tested on Von Kossa staining and Alizarin red staining methods in following 5 mouse VSMCs groups: control group, agonist-CD137 group(recombinant CD137L protein), anti-P38 group (agonist-CD137 group+P38 inhibitor), anti-CD137 group (agonist-CD137 group+CD137 inhibitor),agonist-P38 group(anti-CD137 group+P38 agonist). Results: (1) Compared with the control group, the fluorescence intensity of α-SMA was lower in the agonist-CD137 group(2.79±0.25 vs. 5.42±0.47,P<0.05), while the fluorescence intensity of OPN was higher(4.91±0.23 vs. 1.63±0.26, P<0.05). The fluorescence intensity of α-SMA was partly recovered after adding P38 inhibitor(4.48±0.27 vs. 2.79±0.25,P<0.05),but it was still lower than the control group (4.48±0.27 vs. 5.42±0.47, P<0.05),the fluorescence intensity of OPN decreased(2.66±0.15 vs. 4.91±0.23,P<0.05),but it was still higher than that in the control group (2.66±0.15 vs. 1.63±0.26,P<0.05).The fluorescence intensity of α-SMA and OPN(5.32±0.67 vs. 5.42±0.47,1.82±0.30 vs.1.63±0.26,both P>0.05) was similar between the control group and single anti-P38 group.(2) Compared with the control group, the protein level of p-P38(4.15±0.24 vs. 3.48±0.26, P<0.05), OPN(2.43±0.21 vs. 1.53±0.08, P<0.05), RUNX-2(3.20±0.23 vs. 1.13±0.10, P<0.05) was significantly increased in agonist-CD137 group,the above effects were blocked by adding specific P38 inhibitor SB203580(1.16±0.12 vs. 4.15±0.24, 0.50±0.02 vs. 2.43±0.21,and 1.74±0.14 vs. 3.20±0.23,all P<0.05);the protein level of p-P38(2.93±0.60 vs. 3.48±0.26,P>0.05),OPN (1.4±0.64 vs. 1.53±0.08,P>0.05),RUNX-2(1.26±0.26 vs.1.13±0.10, P>0.05) was similar between single anti-P38 group and the control group. (3) Compared with the control group, the mRNA level of OPN (1.51±0.34 vs. 1, P<0.05) and RUNX-2(2.67±0.19 vs. 1, P<0.05) was significantly upregulated in agonist-CD137 group, and these effects were blocked by adding specific P38 inhibitor SB203580(0.33±0.14 vs. 1 and 0.45±0.03 vs. 1,P<0.05);the mRNA level of OPN (1.05±0.09 vs. 1, P>0.05) and RUNX-2(1.18±0.10 vs. 1, P>0.05) was similar between the single anti-P38 group and the control group.(4) Compared with the control group,the ALP activity and calcium concentration(2.40±0.25 vs. 1.40±0.21,5.51±0.33 vs. 3.15±0.31,both P<0.05) were significantly increased in agonist-CD137 group,while the effects could be blocked by adding specific P38 inhibitor SB203580((1.99±0.07) king unit/gprot vs. (2.40±0.25) king unit/gprot, (3.74±0.20) mmol/gprot vs. (5.51±0.33) mmol/gprot, both P<0.05).The ALP activity and calcium concentration was similar between single anti-P38 group and the control group((1.60±0.25) king unit/gprot vs. (1.40±0.21)king unit/gprot, (2.66±0.28) mmol/gprot vs. (3.15±0.31) mmol/gprot, both P>0.05). (5) Compared with the control group,the calcification of VSMCs in the agonist-CD137 group was significantly increased,while the calcification in the anti-P38 group was significantly reduced.Compared with the agonist-CD137 group,the level of calcification in the anti-CD137 group was obviously increased,and the calcification in the agonist-P38 group was significantly higher than that in the anti-CD137 group and the control group. Conclusion These findings suggest that CD137-CD137L signaling may regulate VSMCs calcification via modulating P38 pathway.

Aim To investigate whether necroptosis mediates chemical hypoxia-induced HT22 mouse hippocampal cell injury and inflammation.Methods HT22 hippocampal cells were exposed to cobalt chloride (CoCl2) to establish a model of the chemical hypoxia-induced injury and inflammation.The expression level of RIP3 (an index of necroptosis) was determined by Western blot.Cell counter kit-8 (CCK-8) assay was used to test the cell viability.Lactate dehydrogenase (LDH) activity in the culture medium was measured with commercial kits.Mitochondrial membrane potential (MMP) was examined by rhodamine123 staining followed by photofluorography.The intracellular level of reactive oxygen species (ROS) was detected by 2', 7'-dichlorfluorescein-diacetate (DCFH-DA) staining followed by photofluorography.The secretion levels of interleukin-1β (IL-1β) and tumor necrosis factor-a (TNF-α) were measured by ELISA.Results Treatment of HT22 hippocampal cells with 600 μmol·L-1 CoCl2 for 36 h markedly induced cytotoxicity, leading to a decrease in cell viability to (52.0±2.65) % , indicating that chemical hypoxia-induced cellular injury model was successfully set up.Besides, CoCl2 induced considerable injuries and inflammation, evidenced by increases in LDH activity, ROS production, MMP loss, as well as the secretion levels of IL-1β and TNF-α.Co-treatment of the cells with 40~100 μmol·L-1 Nec-1 (a specific inhibitor of necroptosis) and CoCl2 markedly attenuated the decrease in viability induced by CoCl2, reaching the best anti-cytotoxicity inhibitory effect at 80 μmol·L-1.Meanwhile, the co-treatment with 80 μmol·L-1 Nec-1 blocked the above injuries and inflammatory response induced by CoCl2.In addition, treatment of HT22 hippocampal cells for 6~48 h up-regulated the expression of RIP3, and Nec-1 alleviated the up-regulation of RIP3 expression level induced by CoCl2.Conclusion Necroptosis mediates chemical hypoxia-induced HT22 hippocampal cell injury and inflammation.