Development and validation of a custom panel including 114 InDels using massively parallel sequencing for forensic application.

Insertion/deletion polymorphisms (InDels) have particular characteristics, such as a relatively low mutation rate, small amplicon size, and no stutter artifacts when genotyped via the capillary electrophoresis platform. It would be an important complementary tool for individual identification and certain kinship analyses. At present, massively parallel sequencing (MPS) has shown excellent application value in forensic studies. Therefore, in this study, we developed a custom MPS InDel panel that contains 114 InDels [77 autosomal InDels (A-InDels), 32 X-chromosomal InDels (X-InDels), and 5 Y-chromosomal InDels) based on previous studies. To assess this panel's performance, several validation experiments were performed, including sensitivity, inhibitor, degraded DNA testing, species specificity, concordance, repeatability, case-type samples, and population studies. The results showed that the lowest DNA input was 0.25 ng. All genotypes were obtained in the presence of 80 ng/µL humic acid, 2000 µmol/L calcium, 3000 µmol/L EDTA and indigo. In degraded DNA testing, 90% of loci could be detected for 16-day-old formalin-fixed hearts. In addition, this panel has good species specificity. The values of combined power of discrimination and the combined power of exclusion for 77 A-InDels were 1-3.9951 × 10-32 and 1-4.2956 × 10-7 , respectively. The combined mean exclusion chance for 32 X-InDels was 0.99999 in trios and 0.99904 in duos. The validation results indicate that this newly developed MPS multiplex system is a robust tool for forensic applications.

Theabrownin from Pu-erh tea attenuates hypercholesterolemia via modulation of gut microbiota and bile acid metabolism.

Pu-erh tea displays cholesterol-lowering properties, but the underlying mechanism has not been elucidated. Theabrownin is one of the most active and abundant pigments in Pu-erh tea. Here, we show that theabrownin alters the gut microbiota in mice and humans, predominantly suppressing microbes associated with bile-salt hydrolase (BSH) activity. Theabrownin increases the levels of ileal conjugated bile acids (BAs) which, in turn, inhibit the intestinal FXR-FGF15 signaling pathway, resulting in increased hepatic production and fecal excretion of BAs, reduced hepatic cholesterol, and decreased lipogenesis. The inhibition of intestinal FXR-FGF15 signaling is accompanied by increased gene expression of enzymes in the alternative BA synthetic pathway, production of hepatic chenodeoxycholic acid, activation of hepatic FXR, and hepatic lipolysis. Our results shed light into the mechanisms behind the cholesterol- and lipid-lowering effects of Pu-erh tea, and suggest that decreased intestinal BSH microbes and/or decreased FXR-FGF15 signaling may be potential anti-hypercholesterolemia and anti-hyperlipidemia therapies.

Effects of extracts from Chuanwu (Aconitum Carmichaelii) and Banxia (Rhizoma Pinelliae) on excisional wound healing in a rat's model.

OBJEVTIVE: To investigate the effects of extracts from Chuanwu (Aconitum Carmichaelii) and Banxia (Rhizoma Pinelliae) on the excisional wound healing in a rat's model. METHODS: Rats were performed a surgical lesion with a 2.0 cm resected tissue in the dorsal fascia. Following, animals were randomly divided into model group, YNB group and APE group those were respectively treated with saline, 1 mg/mL of Yunnan Baiyao and 1 mg/mL of Chuanwu (Aconitum Carmichaelii) and Banxia (Rhizoma Pinelliae) extracts. Wound contractions in days 0, 3, 7, 14 and 21 were calculated by an image analyzer. Histological analysis was analyzed using hematoxilin and eosin. Levels of tumor necrosis factor α (TNF-α), interleukin-2 (IL-2), IL-4, IL-10, transforming growth factor-ß1 (TGF-ß1) and basic fibroblast growth factor (bFGF) were determined by real-time quantitative PCR. RESULTS: Compared with that of YNB group and APE gtoup, the skin of rats showed poor re-modeling and re-epithelization characterized by a significant decrease of neovascularization, epithelialization and fibroblast in the model group. In the APE group, levels of TNF-α and IL-2 were significantly down-regulated and IL-4 and IL-10 significantly up-regulated in contrasted with that of model group. In addition, levels of TGF-ß1 and bFGF in the APE group were significantly induced compared with that of model group. CONCLUSION: The results suggest that the extracts from Chuanwu (Aconitum Carmichaelii) and Banxia (Rhizoma Pinelliae) promote wound healing in the rats, which is associated with enhancing anti-inflammatory ability and inducing fibroblast formation.

Effect of marine collagen peptides on long bone development in growing rats.

BACKGROUND: Nutritional factors play an important role in skeletal development during growth. However, the absorption rates of many bone-related minerals are relatively low from traditional diets, especially for people in Asia and Africa. In the present study, the effect of marine collagen peptides (MCP) derived from chum salmon (Oncorhynchus keta) skin on the development of femurs in growing rats of both sexes was investigated. RESULTS: MCP supplementation could increase the size, mineral density, dry weight, ash weight, content of most minerals and both stiffness and toughness of femurs in male growing rats. However, such effects were milder in female rats. The reported increase in mineral density was likely to be related to increased osteoblast activity rather than a decreased rate of bone resorption, as an increase in serum osteocalcin and bone-specific alkaline phosphatase content was observed, but no significant difference in serum N-terminal telopeptide of type I collagen compared with control was found. CONCLUSION: MCP supplementation could promote the development of long bones in growing male rats.