Optimization and Developmental Validation of 38-plex InDel Panel for Ancestry Inference / 法医学杂志

OBJECTIVES@#The previously established 38-plex InDel system was optimized and its performance was validated according to the Scientific Working Group on DNA Analysis Method (SWGDAM) application guidelines. The ancestry inference accuracy of individuals from East Asian, European, African and mixed populations was verified.@*METHODS@#DNA standard sample 9947A was used as the template to establish the optimal amplification conditions by adjusting primer balance, Mg2+ final concentration and optimizing PCR thermal cycle parameters and amplification volume. The allelic dropout, nonspecific amplification and whether the origin of the inferred samples matched the known information were compared to evaluate the performance of this system.@*RESULTS@#The optimal dosage of this system was 0.125-2 ng DNA template. The results of InDel typing were accurate, the amplification equilibrium was good, and the species specificity was good. This system showed certain tolerance to DNA samples including the inhibitor such as hemoglobin (≤80 μmol/L), indigo (≤40 mmol/L), calcium ion (≤1.0 mmol/L), and humic acid (≤90 ng/μL). The system enabled the direct amplification of DNA from saliva and blood on filter paper, and the results of ethnic inference were accurate. The system successfully detected the mixed DNA sample from two individuals. The test results of the system for common biological materials in practical cases were accurate.@*CONCLUSIONS@#The results of the 38-plex InDel system are accurate and reliable, and the performance of the system meets the requirement of the SWGDAM guidelines. This system can accurately differentiate the ancestry origins of individuals from African, European, East Asian, and Eurasian populations and can be implemented in forensic practice.

Guiqi Dingnian Prescription Delays Senescence of Mouse Mesangial Cells by Regulating JAK2/STAT Pathway Activity / 中国实验方剂学杂志

Objective:To investigate the effects of Guiqi Dingnian prescription （GDP） on the expression of related molecules in Janus tyrosine kinase 2 （JAK2）/signal transducer and activator of transcription （JAK2/STAT） signaling pathway of <italic>D</italic>-galactose （<italic>D</italic>-gal）-induced senescent mesangial cells. Method:The senescent mouse mesangial cells induced by 10 g·L^-1 <italic>D</italic>-gal were continuously treated with 40 mg·L^-1GDP for three days. The senescence of the treated cells was determined by senescence-associated （SA）-<italic>β</italic>-gal staining. The cell cycle was detected by flow cytometry. The cell viability was analyzed using the cell counting kit-8 （CCK-8）. The mRNA expression levels of tumor necrosis factor-<italic>α</italic> （TNF-<italic>α</italic>）， interleukin-6 （IL-6）， nuclear transcription factor-<italic>κ</italic>B （NF-<italic>κ</italic>B）， and IL-1<italic>α</italic> were detected by real-time polymerase chain reaction （Real-time PCR）. The protein expression levels of STAT1， phosphorylated STAT1 （p-STAT1）， STAT3， and p-STAT3 in the JAK2/STAT signaling pathway were determined by Western blot. Result:CCK-8 results showed that the optimal concentration of GDP was 40 mg·L^-1. Compared with the blank group， the positive rate of SA-<italic>β</italic>-gal in the model group was significantly higher（<italic>P</italic><0.01）， the percentage of cells in G₀/G₁ phase was significantly increased（<italic>P</italic><0.05）， the percentage of cells in G₂/M and S phase was significantly decreased（<italic>P</italic><0.01）. The mRNA expression levels of TNF-<italic>α</italic>，IL-6，NF-<italic>κ</italic>B and IL-1<italic>α </italic>were significantly increased（<italic>P</italic><0.01）. Compared with the model group， the model + GDP group exhibited significantly decreased SA-<italic>β</italic>-gal-positive cells （<italic>P</italic><0.05）， reduced cells in the G₀/G₁ phase （<italic>P</italic><0.05）， increased cells in the G₂/M and S phases （<italic>P</italic><0.01）， and down-regulated TNF-<italic>α</italic>， IL-6， NF-<italic>κ</italic>B， and IL-1<italic>α </italic>mRNA expression （<italic>P</italic><0.05） and STAT1， p-STAT1， STAT3， and p-STAT3 protein expression （<italic>P</italic><0.05）. Conclusion:GDP delays the senescence of mouse mesangial cells possibly by down-regulating the expression of related molecules in the JAK2/STAT pathway.

The impact of sodium aescinate on acute lung injury induced by oleic acid in rats.

Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are associated with high rates of morbidity and mortality. Currently, several surfactant or anti-inflammatory drugs are under test as treatments for ALI. Sodium aescinate (SA) has been shown to exert anti-inflammatory and antiedematous effects. In the present work, the authors explored the effects of SA and the possible mechanisms of SA action in rats with ALI induced by oleic acid (OA) administration. Eight groups of rats received infusions of normal saline (NS) or OA. Rats exposed to OA were pretreated with 1 mg/kg of SA, or posttreated with SA at low (1 mg/kg), medium (2 mg/kg), or high (6 mg/kg) dose; a positive-control group received methylprednisolone. The pressure of oxygen in arterial blood (P(O(2))) levels, the pulmonary wet/dry weight (W/D) ratios, and indices of quantitative assessment (IQA) of histological lung injury were obtained 2 or 6 hours after OA injection (0.1 mL/kg, intravenously). The levels of superoxide dismutase (SOD), malondialdehyde (MDA), matrix metalloproteinase gelatinase B (MMP-9), and tissue inhibitor of metalloproteinase (TIMP-1) in both plasma and lung tissue were also determined. Both pre- and posttreatment with SA improved OA-induced pulmonary injury, increased P(O(2)) and SOD values, lowered IQA scores, and decreased the lung W/D ratio and MDA and MMP-9 levels in plasma and lung tissue. SA appeared to abrogate OA-induced ALI by modulating the levels of SOD, MDA, and MMP-9 in plasma and lung tissue.

Effects of cocaine on activities of ATPase, LDH and SDH in mouse splenocytes / 法医学杂志

OBJECTIVE@#To examine the effects of cocaine on the activities of ATPase, LDH and SDH in cultured mouse splenocytes in vitro.@*METHODS@#The ATPase, LDH and SDH activities in mouse splenocytes were detected at day 7 after continuous culturing the mouse cells exposed to cocaine hydrochloride in final concentration of 10, 20 and 100 microg/mL in vitro.@*RESULTS@#The activities of ATPase, LDH and SDH in mouse splenocytes exposed to cocaine hydrochloride in final concentration of 10, 20 and 100 microg/mL were significantly decreased after continuous culturing for 7 days.@*CONCLUSION@#The present study demonstrated that cocaine could inhibit the activities of ATPase, LDH and SDH in cultured splenocytes in vitro.