A novel 193-plex MPS panel integrating STRs and SNPs highlights the application value of forensic genetics in individual identification and paternity testing.

Massively parallel sequencing (MPS) has emerged as a promising technology for targeting multiple genetic loci simultaneously in forensic genetics. Here, a novel 193-plex panel was designed to target 28 A-STRs, 41 Y-STRs, 21 X-STRs, 3 sex-identified loci, and 100 A-SNPs by employing a single-end 400 bp sequencing strategy on the MGISEQ-2000™ platform. In the present study, a series of validations and sequencing of 1642 population samples were performed to evaluate the overall performance of the MPS-based panel and its practicality in forensic application according to the SWGDAM guidelines. In general, the 193-plex markers in our panel showed good performance in terms of species specificity, stability, and repeatability. Compared to commercial kits, this panel achieved 100% concordance for standard gDNA and 99.87% concordance for 14,560 population genotypes. Moreover, this panel detected 100% of the loci from 0.5 ng of DNA template and all unique alleles at a 1:4 DNA mixture ratio (0.2 ng minor contributor), and the applicability of the proposed approach for tracing and degrading DNA was further supported by case samples. In addition, several forensic parameters of STRs and SNPs were calculated in a population study. High CPE and CPD values greater than 0.9999999 were clearly demonstrated and these results could be useful references for the application of this panel in individual identification and paternity testing. Overall, this 193-plex MPS panel has been shown to be a reliable, repeatable, robust, inexpensive, and powerful tool sufficient for forensic practice.

Valorization of Fish Processing By-Products: Microstructural, Rheological, Functional, and Properties of Silver Carp Skin Type I Collagen.

The objective of this study was to develop aquatic collagen production from fish processing by-product skin as a possible alternative to terrestrial sources. Silver carp skin collagen (SCSC) was isolated and identified as type I collagen, and LC-MS/MS analysis confirmed the SCSC as Hypophthalmichthys molitrix type I collagen, where the yield of SCSC was 40.35 ± 0.63% (dry basis weight). The thermal denaturation temperature (Td) value of SCSC was 30.37 °C, which was superior to the collagen of deep-sea fish and freshwater fish. Notably, SCSC had higher thermal stability than human placental collagen, and the rheological experiments showed that the SCSC was a shear-thinning pseudoplastic fluid. Moreover, SCSC was functionally superior to some other collagens from terrestrial sources, such as sheep, chicken cartilage, and pig skin collagen. Additionally, SCSC could provide a suitable environment for MC3T3-E1 cell growth and maintain normal cellular morphology. These results indicated that SCSC could be used for further applications in food, cosmetics, and biomedical fields.

Soluble Flt-1 in AMI Patients Serum Inhibits Angiogenesis of Endothelial Progenitor Cells by Suppressing Akt and Erk's Activity.

In acute myocardial infarction (AMI), endothelial progenitor cells (EPCs) are essential for the recovery of collateral circulation via angiogenesis. Clinical research has shown that the poor prognosis of the patients with AMI is closely associated with the cell quantity and function of EPCs. Whether there are differences in the biological features of EPCs from AMI patients and healthy subjects is worth exploring. In this study, EPCs were isolated from human peripheral blood and identified as late-stage EPCs by flow cytometry, immunofluorescence, and blood vessel formation assay. Compared to healthy subjects, AMI patients had more EPCs in the peripheral blood compared to healthy subjects. In addition, EPCs from AMI patients exhibited higher migration ability in the transwell assay compared to EPCs from healthy subjects. However, no difference in the angiogenesis of EPCs was observed between AMI patients and healthy subjects. Further studies revealed that soluble vascular endothelial growth factor receptor 1 (sFlt-1) in the serum of AMI patients was involved in the inhibition of EPCs angiogenesis by suppressing the Akt and Erk pathways. In conclusion, this study demonstrated that elevated serum sFlt-1 inhibits angiogenesis of EPC in AMI patients. Our findings uncover a pathogenic role of sFlt-1 in AMI.

Blocking the death checkpoint protein TRAIL improves cardiac function after myocardial infarction in monkeys, pigs, and rats.

Myocardial infarction (MI) is a leading cause of death worldwide for which there is no cure. Although cardiac cell death is a well-recognized pathological mechanism of MI, therapeutic blockade of cell death to treat MI is not straightforward. Death receptor 5 (DR5) and its ligand TRAIL [tumor necrosis factor (TNF)-related apoptosis-inducing ligand] are up-regulated in MI, but their roles in pathological remodeling are unknown. Here, we report that blocking TRAIL with a soluble DR5 immunoglobulin fusion protein diminished MI by preventing cardiac cell death and inflammation in rats, pigs, and monkeys. Mechanistically, TRAIL induced the death of cardiomyocytes and recruited and activated leukocytes, directly and indirectly causing cardiac injury. Transcriptome profiling revealed increased expression of inflammatory cytokines in infarcted heart tissue, which was markedly reduced by TRAIL blockade. Together, our findings indicate that TRAIL mediates MI directly by targeting cardiomyocytes and indirectly by affecting myeloid cells, supporting TRAIL blockade as a potential therapeutic strategy for treating MI.

Improvement of the production of an Arctic bacterial exopolysaccharide with protective effect on human skin cells against UV-induced oxidative stress.

Although microbial exopolysaccharides (EPSs) are applied in different fields, no EPS has been used to protect human skin cells against UV-induced oxidative stress. The EPS produced by the Arctic bacterium Polaribacter sp. SM1127 has high moisture-retention ability and antioxidant activity, suggesting its good industrial potentials. In this study, we improved the EPS production of SM1127 and evaluated its protective effect on human dermal fibroblasts (HDFs) against UV-induced oxidative stress. With glucose as carbon source, the EPS yield was increased from 2.11 to 6.12 g/L by optimizing the fermentation conditions using response surface methodology. To lower the fermentation cost and decrease corrosive speed in stainless steel tanks, whole sugar, whose price is only 8% of that of glucose, was used to replace glucose and NaCl concentration was reduced to 4 g/L in the medium. With the optimized conditions, fed-batch fermentation in a 5-L bioreactor was conducted, and the EPS production reached 19.25 g/L, which represents the highest one reported for a polar microorganism. Moreover, SM1127 EPS could maintain the cell viability and integrity of HDFs under UV-B radiation, probably via decreasing intracellular reactive oxygen species level and increasing intracellular glutathione content and superoxide dismutase activity. Therefore, SM1127 EPS has significant protective effect on HDFs against UV-induced oxidative stress, suggesting its potential to be used in preventing photoaging and photocarcinogenesis. Altogether, this study lays a good foundation for the industrialization of SM1127 EPS, which has promising potential to be used in cosmetics and medical fields.

Downregulated miR-203 attenuates IL-ß, IL-6, and TNF-α activation in TRAF6-treated human renal mesangial and tubular epithelial cells.

Circulating microRNAs (miRNAs) are attracting major interest as novel non-invasive biomarkers for human autoimmune diseases including lupus nephritis (LN). A previous study showed that altered miR-203 expression may provide highly diagnostic for systemic lupus erythematosus. However, whether miR-203 is a diagnostic biomarker for LN is still unknown. In the present research, serum samples from 35 cases of active LN patients, 58 cases of inactive LN patients, and 74 cases of healthy volunteers were collected to analyze the expression profiles of miR-203 by qRT-PCR. The serum concentration of complement component 3 (C3) and complement component 4 (C4) was detected using nephelometry method. The expression of inflammatory cytokines, including interleukin 1 beta (IL-1ß), interleukin 6 (IL-6), and tumor necrosis factor α (TNF-α), were analyzed using enzyme-linked immunosorbent assay (ELISA). The effect of miR-203 overexpression on the TNF receptor associated factor 6 (TRAF6)-induced inflammation of human renal mesangial cells (HRMCs) and human renal tubular epithelial cell line (HK-2) were evaluated. Results showed that miR-203 in serum of active LN patients was significantly down-regulated when compared with serum from inactive LN patients and healthy volunteers. Receiver operating curve (ROC) showed that decreased circulating miR-203 was a significant diagnostic biomarker for active LN patients, with an area under curve (AUC) of 0.974; sensitivity was 85.79%, and specificity was 89.40%. Significant downregulation of C3 and C4, and obvious upregulation of IL-ß, IL-6, and TNF-α, was observed in serum of active LN patients. Furthermore, circulating miR-203 expression was positively correlated with the serum concentrations of C3 and C4, and negatively correlated with the serum expression of IL-1ß, IL-6, and TNF-α in active LN patients. In addition, transfection of HRMCs and HK-2 cells with miR-203 mimics could suppress TRAF6-induced IL-ß, IL-6, or TNF-α expression compared to cells treated with the mimics control group. In summary, decreased circulating miR-203 might be a candidate diagnostic biomarker for human active LN, and it attenuated IL-ß, IL-6, and TNF-α activation in TRAF6-treated HRMCs and HK-2 cells.

MicroRNA-124 represents a novel diagnostic marker in human lupus nephritis and plays an inhibitory effect on the growth and inflammation of renal mesangial cells by targeting TRAF6.

microRNAs (miRs) are short non-coding RNAs that function as guide molecules in RNA silencing by inducing mRNA degradation or blocking protein translation. Increasing evidence has shown that miRNAs play an important role in regulating the pathological process of lupus nephritis (LN), but the precise role of miR-124 in LN is still unknown. Here, we found that miR-124 expression is significantly reduced in patients with active LN compared with those patients with non-active LN and the absence of LN. Additionally, the miR-124 level was negatively correlated with serum IL-1ß, IL-6, TNF-α, and TRAF6 mRNA expressions in active LN patients. Receiver operating characteristic and logistic regression analyses revealed miR-124 is a significant diagnostic biomarker for active LN. Furthermore, transfection of the miR-124 mimic into human renal mesangial cells (HRMCs) resulted in significantly reduced cell proliferation, induced cell apoptosis, and decreased synthesis of inflammatory factors. Moreover, a dual luciferase assay showed that TRAF6 was a direct target of miR-124, and the expression of TRAF6 was suppressed by miR-124 through direct binding to the 3'-UTR of mRNA. Mechanistic studies demonstrated that the over-expression of TRAF6 could abrogate miR-124-related effects on cell proliferation, apoptosis and the synthesis of inflammatory factors in HRMCs. Taken together, these findings indicate that downregulated miR-124 represents a novel diagnostic marker in human LN and plays an inhibitory effect on the growth and inflammation of renal mesangial cells by targeting TRAF6.

Growth differentiation factor 11 ameliorates experimental colitis by inhibiting NLRP3 inflammasome activation.

Growth differentiation factor 11 (GDF11) has an anti-inflammatory effect in the mouse model of atherosclerosis and Alzheimer's disease, but how GDF11 regulates intestinal inflammation during ulcerative colitis (UC) is poorly defined. The Nod-like receptor family pyrin domain-1 containing 3 (NLRP3) inflammasome is closely associated with intestinal inflammation because of its ability to increase IL-1ß secretion. Our aim is to determine whether GDF11 has an effect on attenuating experimental colitis in mice. In this study, using a dextran sodium sulfate (DSS)-induced acute colitis mouse model, we reported that GDF11 treatment attenuated loss of body weight, the severity of the disease activity index, shortening of the colon, and histological changes in the colon. GDF11 remarkably suppressed IL-1ß secretion and NLRP3 inflammasome activation in colon samples and RAW 264.7 cells, such as the levels of NLRP3 and activated caspase-1. Furthermore, we found that GDF11 inhibited NLRP3 inflammasome activation by downregulating the Toll-like receptor 4/NF-κB p65 pathway and reactive oxygen species production via the typical Smad2/3 pathway. Thus, our research shows that GDF11 alleviates DSS-induced colitis by inhibiting NLRP3 inflammasome activation, providing some basis for its potential use in the treatment of UC. NEW & NOTEWORTHY Here, we identify a new role for growth differentiation factor 11 (GDF11), which ameliorates dextran sodium sulfate-induced acute colitis. Meanwhile, we discover a new phenomenon of GDF11 inhibiting IL-1ß secretion and Nod-like receptor family pyrin domain-1 containing 3 (NLRP3) inflammasome activation. These findings reveal that GDF11 is a new potential candidate for the treatment of ulcerative colitis patients with a hyperactive NLRP3 inflammasome.