Ancestry and phenotype predictions from touch DNA using massively parallel sequencing

International Journal of Legal Medicine - Direct PCR can be used to successfully generate full STR profiles from DNA present on the surface of objects. STR profiles are only of use in cases where a...     

针灸治疗胃脘痛的古代文献研究

胃脘痛是临床常见病之一,针灸治疗胃脘痛具有肯定的疗效。中医古代文献中关于针灸治疗胃脘痛的记载内容丰富,但缺乏系统整理。检索古代医学相关文献中针灸治疗胃脘痛相关条目,并对所涉及腧穴和信息加以归纳、分析与研究,通过探究针灸治疗胃脘痛的临床应用。     

Asiaticoside induces osteogenic differentiation of human periodontal ligament cells through the Wnt pathway

1 Background

Asiaticoside is a compound isolated from Herb Centella asiatica, which has been shown to promote osteogenic differentiation of human periodontal ligament (hPDL) cells. This study investigated the molecular mechanism underlying the asiaticoside‐induced osteogenic differentiation of hPDL cells.

2 Methods

hPDL cells were incubated with various concentrations of asiaticoside to test cell viability by MTT assay. The mRNA expression levels were analyzed by using quantitative real‐time polymerase chain reaction (PCR). Osteogenic differentiation was determined by alkaline phosphatase activity assay and alizarin red staining. The subcellular localization of β‐catenin was analyzed by both immunofluorescence and western blot.

3 Results

The results showed that asiaticoside had no effect on the cell viability at any of the tested concentrations. Real‐time PCR revealed that osterix (OSX) and dentin matrix protein1 (DMP1) mRNA were significantly enhanced by asiaticoside treatment. Alkaline phosphatase activity and in vitro mineralization were also significantly induced. Interestingly, asiaticoside dose‐dependently increased WNT3A mRNA expression, but not WNT5A and WNT10B. The activation of Wnt signaling was shown to result in nuclear accumulation of β‐catenin as evaluated by immunofluorescence staining and western blot analysis. Pre‐treatment with recombinant human Dickkopf1 (rhDKK1) inhibited asiaticoside‐induced β‐catenin nuclear translocation and osteoblast marker gene expression. Moreover, rhDKK1 attenuated asiaticoside‐induced DMP1 protein expression.

4 Conclusion

The data demonstrate that asiaticoside induces osteogenic differentiation of hPDL cells by activating the Wnt/β‐catenin signaling pathway. The findings suggest that asiaticoside could be used as a novel therapeutic drug for periodontal tissue regeneration.     

Locating DNA within fingermarks using fluorescent in situ detection; a collaboration between ESR and Flinders University

ABSTRACT

A pilot study on the detection of DNA in fingermarks using fluorescent in situ detection after different time periods since hand washing was undertaken by Flinders University. Collaboration was sought to show the ability to obtain similar results within different laboratories. The Institute of Environmental Science and Research (ESR) was involved in this inter-laboratory study in collaboration with Flinders University. The newly developed method involves the use of Diamond Nucleic Acid dye for the staining of fingermarks (20× concentration in 75% ethanol) using a hand-held microscope (Dino-Lite Edge Digital Microscope). Fingermarks were deposited by volunteers onto glass slides at varying time intervals after hand washing (2, 5, 15, 30, 60 and 180 minutes). The amount of cellular debris was calculated by counting the fluorescent dots present in three fields of view and estimating the amount of transferred cellular material for each fingermark. This article will outline the results obtained from ESR and how they compare with results already collated from Flinders University.     

Loss of cyclin E requirement in cell growth of an oral squamous cell carcinoma cell line implies deregulation of its downstream pathway

Cyclin E and Cdk2 have been shown to play an important role in G1/S transition of the cell cycle. Two E-type cyclins (E1 and E2) have been identified to date and share functionally similarities. Upregulation of these cyclins has been observed frequently in human cancers. We examined the expression profile of cyclin E1 and E2 in cell lines derived from human oral squamous cell carcinoma (SCC), and found that the expression of cyclin E1 protein was hardly detected in HSC-2 cells. Although cyclin E2 was abundantly expressed, histone H1 kinase activities of both E-type cyclins were virtually undetectable in this cell line. Inhibition of cyclin E1, but not that of E2, by using vectors expressing antisense-oriented their cDNAs induced drastic growth suppression on HOC313 cells that express both E-type cyclins. Inhibition of neither cyclin E1 nor E2 suppressed the growth of HSC-2 cells, and compensatory elevation of cyclin E1 was not evident in cyclin E2-inhibited HSC-2 cells. In contrast, HSC-2 cells expressed cyclin D1 and hyperphosphorylated forms of Rb family proteins, and were arrested in G1 by overexpression of p16(INK4), a specific inhibitor against D-type cyclin activity. These results indicate that HSC-2 cells lost proper growth control specifically mediated by cyclin E and suggest that deregulation of its downstream pathway may contribute to tumorigenesis of oral SCC.     

Role of cyclin D1 cytoplasmic sequestration in the survival of postmitotic neurons

Cyclin D-dependent kinases phosphorylate the retinoblastoma (Rb) protein and play a critical role in neuronal cell cycle control and apoptosis. Here we show that cyclin D1 became predominantly cytoplasmic as primary cortical progenitor cells underwent cell cycle withdrawal and terminal differentiation. Furthermore, ectopically expressed cyclin D1 sequestered in the cytoplasm of postmitotic neurons, whereas it efficiently entered the nucleus of proliferating progenitor cells. Cytoplasmic cyclin D1 were complexed with cyclin-dependent kinase 4 (CDK4), and also with CDK inhibitors, p27(Kip)(I) or p21(Cip)(I), which positively regulate assembly and nuclear accumulation of the cyclin D1-CDK4 complex. Although overexpression of p21(Cip)(I) promoted cyclin D1 nuclear localization, inhibition of either glycogen synthase kinase 3beta- or CRM1-mediated cyclin D1 nuclear export did not, suggesting that the inhibition of its nuclear import, rather than the acceleration of nuclear export, contributes to cytoplasmic sequestration of cyclin D1 in postmitotic neurons. In differentiated progenitor cells, nuclear localization of ectopic cyclin D1 induced apoptosis, and the DNA-damaging compound camptothecin caused nuclear accumulation of endogenous cyclin D1, accompanied by Rb phosphorylation. These results indicate that nuclear accumulation of cyclin D1 is inhibited in postmitotic neurons and suggest a role of its subcellular localization in neuronal death and survival.     

Short cervix detection in pregnant women by transabdominal sonography with post-void technique

Purpose

This study aimed to evaluate the diagnostic properties of transabdominal sonography with the post-void technique for cervical length measurement.

Methods

This study was a prospective cohort study. The inclusion criteria were pregnant women aged 18–40 years with gestational age of 18–23 completed weeks. Transabdominal sonography with vertical bladder depth of less than 5 cm and transvaginal cervical length measurements were carried out by a single experienced sonographer.

Results

There were 307 eligible pregnant women. The mean age of all subjects was 29.0 years. The mean cervical length obtained through transabdominal and transvaginal measurement was 3.33 and 3.47 cm, respectively. Ten patients (3.3 %) were identified as having a short cervix using transvaginal sonography, and 12 patients (3.9 %) were identified using transabdominal sonography.

Conclusion

Transabdominal sonography with vertical bladder depth of less than 5 cm performed better compared with transvaginal sonography. It may not be necessary to perform transvaginal sonography if transabdominal sonography reveals the cervical length to be more than 2.5 cm.

     

Detection of cellular material in lip-prints

We report on the visualization of cellular material within lip-prints using Diamond™ dye (DD). The transfer of cellular material via the lips can occur in cases of contact with food or drinking items as well as cases of alleged sexual assault involving oral contact. DD can effectively detect cellular material transferred by touch. Here we investigate if lip-prints can be detected and whether there is consistency within, or variability between, a person’s propensity to shed cells within lip-prints. Ten volunteers were asked to press their lips against a glass slide with medium pressure for 15 s after not eating or drinking for at least 30 min. Both upper and lower lips were observed, and all tests were performed in five replicates, giving in total 900 observed areas. Consistency in the amount of cellular material deposited by lip-prints for each of the 10 individuals was observed, with each individual being associated with a ‘lip shedder’ status between the extremes of heavy and light. The majority of females shed more cells than the majority of males. No correlation was observed between the lip-prints shedder-status compared to deposition of cellular material from a thumb. Further, no correlation was observed between lip morphology and the ‘lip shedder’ status. Visualization of cellular material was not affected by lip-balm but was adversely affected by cosmetics such as lipstick. This technique demonstrates the visualization of deposited cells from parts of the body other than fingers and how cellular material can be visualized allowing targeted collection of DNA.     

Pressure induces interleukin-6 expression via the P2Y6 receptor in human dental pulp cells

Background and objective

An increase in intrapulpal pressure occurs during inflammation and restorative procedures; however, the role of the pressure on human dental pulp cell (HDPC) is not yet clarified. In this study, the effect of pressure on interleukin-6 (IL-6) expression of HDPCs was examined.

Design

HDPCs were applied with pressure (0.7–1.4 g/cm²). The level of IL-6 mRNA and protein release was determined by RT-PCR and ELISA, respectively. The signalling pathways were investigated using inhibitors, antagonists, and small interfering RNA.

Results

The results showed that pressure up-regulated IL-6 mRNA expression and protein release in a time- and dose-dependent manner. The implication of P2Y receptor was exhibited by a significant inhibition of pressure-induced IL-6 expression by suramin, an antagonist for the non-specific purinergic receptor family. Using loss of function experiments, we showed MRS2578 (a specific P2Y6 antagonist), as well as P2Y6 small interfering RNA, abolished pressure-induced IL-6, whilst MRS2179 (a specific P2Y1 antagonist) and NF449 (a P2X1, P2X3, P2Y1, and P2Y2 antagonist) had no effect. Finally, we demonstrated that either the conditioned medium collected from pressurised dental pulp cells or addition of UDP, a selective agonist of P2Y6, up-regulated IL-6 expression in HDPCs.

Conclusions

These results indicate that pressure could induce IL-6 expression through the P2Y6 receptor in HDPCs, leading to a new insight of the role of pressure on cytokine release during pulpal inflammatory process.     

Prevention of cyclin D1 nuclear localization in terminally differentiated neurons

Terminally differentiated neurons irreversibly withdraw from the cell cycle. The mechanisms governing the activity of cyclin D 1, a key regulator of the cell cycle, during neuronal cell cycle withdrawal are not fully understood. This study shows that cyclin D 1 became predominantly cytoplasmic in differentiated cortical neurons. Cytoplasmic cyclin D 1 assembled with cyclin dependent kinase 4 (CDK 4), and the CDK inhibitors p21Cip1 and p27Kip1. Although forced expression of p 21 caused cyclin D 1 nuclear accumulation, the inhibition of its nuclear export by inhibiting GSK-3 beta activity had no effect. Furthermore, ectopically expressed cyclin D 1 entered the nucleus of proliferating nervous, but not that of differentiated neurons, whereas ectopic cyclin D 1 in quiescent fibroblasts accumulated in the nucleus and induced cell cycle progression. These results indicate that cyclin D 1 nuclear localization is tightly inhibited in terminally differentiated neurons, and suggest that the regulation of its nuclear import plays a role in neuronal cell cycle withdrawal.