Single-cell transcriptome in silico analysis reveals conserved regulatory programs in macrophages/monocytes of abdominal aortic aneurysm from multiple mouse models and human.

Abdominal aortic aneurysm (AAA) is a life-threatening disease and there is currently a lack of effective treatment to prevent it rupturing. ScRNA-seq studies of AAA are still lacking. In the study, we analyzed the published AAA scRNA-seq datasets from the mouse elastase-induced model, CaCl2 treatment model, Ang II-induced model and human by using bioinformatic approaches and in silico analysis. A total of 26 cell clusters were obtained and 11 cell types were identified from multiple mouse AAA models. Also, the proportion of Mφ/Mo increased in the AAA group and Mφ/Mo was divided into seven subtypes. There were significant differences in transcriptional regulation patterns of Mφ/Mo in different AAA models. The enrichment pathways of upregulated or downregulated genes from Mφ/Mo in the three mouse datasets were different. The actived regulons of Mφ/Mo had strong specificity and the repressed regulons showed high consistency. The co-upregulated genes as well as actived regulons and co-downregulated genes as well as repressed regulons were closely correlated and formed regulatory networks. Mφ/Mo from human AAA dataset was divided into five subtypes. The proportion of three macrophage subpopulations increased but the proportion of two monocyte subpopulations decreased. In the AAA group, the upregulated or downregulated genes of Mφ/Mo were enriched in different pathways. After further analyzing the genes in Mφ/Mo of both mouse and human scRNA-seq datasets, two genes were upregulated in the four datasets, IL-1B and THBS1. In conclusion, in silico analysis of scRNA-seq revealed that Mφ/Mo and their regulatory related genes as well as interaction networks played an important role in the pathogenesis of AAA.

Congenital fibrinogen disorders with repeated thrombosis.

Congenital dysfibrinogenemia is characterized with undetectable or low fibrinogen level by Clauss assay complicated by bleeding and/or thrombosis. These may lead to a diagnostic problem to some clinicians unfamiliar with this disease. We reported a case of congenital dysfibrinogenemia manifested as hemorrhage, repeated thrombosis, low fibrinogen levels through Clauss assay and but normal levels of fibrinogen through PT-derived tests. In conclusion, to patients with thrombosis complicated by decreased fibrinogen level, clinicians and laboratory physicians should be alert to the possibility of congenital dysfibrinogenemia.

Comparison Thigh Skeletal Muscles between Snowboarding Halfpipe Athletes and Healthy Volunteers Using Quantitative Multi-Parameter Magnetic Resonance Imaging at Rest.

BACKGROUND: Magnetic resonance (MR) imaging provides a unique, noninvasive diagnostic platform to quantify the physiological and biochemical variables of skeletal muscle at rest. This study was to investigate the difference in thigh skeletal muscles between snowboarding halfpipe athletes and healthy volunteers via multiparametric MR imaging. METHODS: A comparative study was conducted between 12 healthy volunteers and 14 snowboarding halfpipe athletes. MR scanning targeted the left leg at the level of the proximal thigh on a 3.0T MR system. The measured parameters compared between the two groups included T1, T2, T2* relaxation times, fat fraction (FF), and cross-sectional area (CSA) of the quadriceps femoris and the hamstring muscles. Statistical analysis was carried out using independent sample t-test. Interrater reliability was also assessed with intraclass correlation coefficients (ICCs). RESULTS: It was statistically equivalent between two groups in age, body mass index, thigh circumference, calf circumference, systolic blood pressure, and resting heart rate (all P > 0.05). However, the T1 and T2 values of the hamstring muscles in the athlete group were found to be significantly shorter than those in control group (T1: 1063.3 ± 24.1 ms vs. 1112.0 ± 38.2 ms in biceps femoris, 1050.4 ± 31.2 ms vs. 1095.0 ± 39.5 ms in semitendinosus, 1053.1 ± 31.7 ms vs. 1118.4 ± 40.0 ms in semimembranosus, respectively; T2: 33.4 ± 0.7 ms vs. 36.1 ± 1.9 ms in biceps femoris, 34.6 ± 2.0 ms vs. 37.0 ± 1.9 ms in semitendinosus, 36.9 ± 1.5 ms vs. 38.9 ± 2.4 ms in semimembranosus, respectively; all P < 0.05) although T2* relaxation time was detected with no significant difference. The FF of the hamstring muscles was obviously less than the control group (5.5 ± 1.9% vs. 10.7 ± 4.7%, P < 0.001). In addition, the quadriceps' CSA in the athlete group was substantially larger than the control group (8039.0 ± 1072.3 vs. 6258.2 ± 852.0 mm2, P < 0.001). Interrater reliability was excellent (ICC: 0.758-0.994). CONCLUSION: Multiple MR imaging parameters indicated significant differences between snowboarding halfpipe athletes and healthy volunteers in the thigh skeletal muscles.

[A case-control study on the risk factors for attempted suicide in patients with major depression].

OBJECTIVE: To understand the environmental risk factors on attempted suicide in patients with major depression, and to study the interaction between factors as single nucleotide polymorphism(SNP) of TPH2 gene rs7305115 associated to attempted suicide in major depression. METHODS: Paired case-control study on 215 suicide attempters with major depression (92 male, 123 female) and molecular biological techniques were used to study the relation between TPH2 gene rs7305115 SNP,interrelated environmental factors and the rate of attempted suicide. Controls were paired with cases according to the same gender, similar age (no more than 3 years) and from the same district. RESULTS: There were remarkably significant differences in gene types and gene frequency between case and control groups (P < 0.001). Data from multivariate conditional logistic regression model analysis showed that hopelessness, negative life-events and family history of suicide were relationship of attempted suicide in patients with major depression with OR values as 0.33 (95% CI: 0.22-0.99), 7.68 (95% CI: 5.79-13.74), 6.64 (95% CI: 2.48-11.04), 2.98 (95% CI: 1.17-5.04) respectively. There was no first level interaction between any of the two risk factors. CONCLUSION: Results from the study supported the idea that hopelessness, negative life-events and family history of suicide were risk factors of attempted suicide in major deprbssion while TPH2 gene rs7305115 A/A might be the protective factor.

Alterations of trace elements (Zn, Se, Cu, Fe) and related metalloenzymes in rabbit blood after severe trauma.

To investigate the status of the trace elements (TEs) and related metalloenzymes activities in the injury and repair process after severe trauma, we established a rabbit model of severe trauma whose Injury Severity Score (ISS) was 22. Concentrations of blood selenium (Se) and serum copper (Cu), zinc (Zn), iron (Fe), and ferritin were measured on D0 (before injury), and day (D) 1, D2, D3, D6, D9, D14, D21, D28 after trauma, respectively. The activities of glutathione peroxidase (GPx), Cu/Zn superoxide dismutase (Cu/Zn-SOD), myeloperoxidase (MPO), the contents of lipid peroxidation product malondialdehyde (MDA) and serum biochemical profile were detected synchronously. In addition, the morphologic changes of major organs were observed at different time intervals. Results showed that blood Se and serum Zn, Fe contents decreased significantly within 2 weeks after injury. Serum Cu concentration was significantly reduced on D1 but normalized quickly. Serum ferritin level increased during the first week while following an obvious decrease thereafter. The blood GPx activity dropped markedly from D1 to D6, the serum Cu/Zn-SOD activity decreased on D1 and then increased significantly within 2 weeks, and the blood MPO-positive stained cells increased within a week after trauma and followed by a decrease from D14 to D21. The serum MDA increased significantly on D6. Seven of 34 rabbits died in 4-6 days after injury. Biochemistry values and pathological features revealed these rabbits died of multiple organ dysfunction syndrome (MODS). Our experiment suggested that the circulating TEs status is dramatically modified in response to trauma, which might be a factor in MODS.

Cloning and characterization of a LASS1-GDF1 transcript in rat cerebral cortex: conservation of a bicistronic structure.

LASS1 is the mammalian homologue of yeast longevity-assurance gene 1 (LAG1) which is differentially expressed during the yeast replicative life span and was shown to play a role in determining yeast longevity. Growth/differentiation factor 1 (GDF1) is a transforming growth factor-beta family member that was originally isolated from an mouse embryo cDNA library. GDF1 is expressed specifically in the nervous system and functions in left-right patterning of the mouse embryo. To explore the potential role of LASS1 in rat neuron aging, northern blot analysis for LASS1 mRNA was performed and detected a 2.7 kb transcript in adult rat cerebral cortex. Cloning and sequence analysis revealed that this transcript contains two nonoverlapping open reading frames (ORFs), LASS1 and GDF1, which are quite different to the predicted sequences in GenBank (accession number XM_224734 and XM_224733). The alignment with the rat genome database revealed this transcript matches the sequence of rat chromosome 16 genomic contig (GenBank accession number NW_047470) between nucleotide positions 1935060th and 1919439th, which contains eight exons and seven introns. Multiple sequence analysis revealed high conservation of the two ORFs. The phylogenetic analysis showed very close evolutionary relationship among rat, mouse and human. It raises the possibility that this mRNA may give rise to two different proteins and the conserved bicistronic structure might be of unknown significance.

Reversal MDR in breast carcinoma cells by transfection of ribozyme designed according the secondary structure of mdr1 mRNA.

Multidrug resistance (MDR) is a major obstacle in cancer chemotherapy. The present study aims to investigate whether the ribozyme could reverse MDR in breast carcinoma cells. In this study, two GUC sites (GUC106 and GUC135) on the surface of mdr1 mRNA were selected according to the secondary structure of the 5'-region of mdrl mRNA. The ribozyme gene RZ106 and RZ135 complementary to two sides bases of the target GUC were synthesized and cloned into the plasmid pEGFP -C1 which has EGFP (Enhanced Green Fluorescence Protein) as report gene and Kan/Neo as selection gene. After transfection with the recombinant plasmid and selected by G418, the stable cell clones were produced and used for detection. The alteration of mdr1 mRNA and P-gp in the treated cells was detected by RT-PCR, flow cytometry and Rh123 retention. The reversal efficiency of the drug resistance for adriamycin was determined by MTT assay. The results showed that after transfection with RZ106 and RZ135, the amount of the mdr1 mRNA and P-gp decreased significantly and the efflux function of P-gp was inhibited accordingly. Nine-fold and 16-fold reduction of resistance for adriamycin was observed in the two groups of treated cells. These results suggested that both ribozymes can reverse the MDR phenotype by inhibiting the expression of mdr1 mRNA and P-gp, and the RZ135 showed the better cleavage efficiency. The ribozyme strategy designed according the secondary structure of the target RNA could be a useful therapy for reversal of MDR.