Objective To apply the real-time quantitative PCR assay with the TaqMan Probe to DNA analysis in Forensic science . Methods Using the TaqMan method to quantitate DNA abstracted from variety of biological samples common in Forensic science . Results The quantity of sample DNA can be obtained. Inhibiting factors in the abstracted DNA might be evaluated. And subsequent procedures of STR testing were then directed. Conclusion Real-time quantitative PCR is a necessary technique for DNA analysis in Forensic science.

Objective To investigate the clinical significance of nuclear factor ( NF)-κB activation in peripheral blood mononuclear cells (PBMCs) and the serum levels of correlated inflammatory cytokines in children with infant muggy syndrome(IMS).Methods Blood samples from 100 patients with IMS and those from 32 healthy infants( control group)were detected by ELISA for amount of NF-κB activation in PBMCs and for serum levels of interleukin ( IL ) -17,IL-6,tumor necrosis factor ( TNF ) -α and IL- 10 respectively from Jan 2008 to Jan 2011.At the same time,blood samples from 46 out of the above 100 patients with IMS and those from the 32 controls for positive rate of activation of NF-κB in PBMCs were detected by flow cytometry as well.The relationship between all the data and multiple organ dysfunction syndrome( MODS ) were analyzed respectively.Results As compared with that of control group,the percentage of activated NF-κB in PBMCs in 100 patients with IMS detected by ELISA [ ( 11.042 ± 6.792 ) ％ vs ( 4.528 ± 1.378 ) ％ ] and the positive rate of NF-κB activation in 46 patients with IMS detected by flow cytometry [ ( 28.780 ± 13.820 ) ％ vs (7.078 ±5.395)％ ] were both significantly higher ( P ＜0.01 ).The serum levels of IL-17,IL-6 and IL-10were also significantly higher in patients with IMS than those in control group( P ＜0.01 ).The serum level of TNF-α was higher in patients with IMS than that in control group but without significance( P ＞ 0.05 ).The percentage of activated NF-κB [ ( 14.591 ± 7.626) ％ vs ( 8.576 ± 4.851 ) ％ ],the positive rate of NF-κB activation [ ( 36.087 ± 12.056) ％ vs ( 23.590 ± 11.263 ) ％ ],and the serum levels of IL- 17,IL-6,TNF-α and IL-10 were all significantly higher in IMS patients with MODS than those in IMS patients without MODS ( P ＜ 0.01 ).Conclusion The inflammatory factors of NF-κB activation in PBMCs and the high serum levels of IL-17 and IL-6 are potent to cause inflammatory damage in IMS patients,and the serum level of IL-10 is not able to compensate the damage.The activation of NF-κB and high serum levels of IL-17,IL-6 and TNF-α are correlated with MODS.

MicroRNA has speciifc biological and expression properties in transcription level, which has more potential than genomic DNA in the identification for forensic body fluid, species and degraded crime biological materials. Here we introduce the speciifc research of forensic body lfuid identiifcation, research strategies and applied perspectives with forensic miRNA, expecting to provide the application and study of miRNA analysis for reference.

Air Pollutants, Occupational ; chemistry ; Indium ; analysis ; Mass Spectrometry ; methods ; Occupational Exposure ; Spectrum Analysis ; methods ; Workplace

Objective:To investigate the effect of transforming growth factor (TGF- β) signal in muscle fiber itself during inflammation/immunity response on intramuscular inflammation. Methods:Sixteen wild C57BL/6 mice (wild group) and sixteen mice with skeletal muscle-specific deficiency of T βRⅡ (knock-out group) between 4-8 weeks of age were selected for this study. Acute muscle injury in mice was induced by injection of myotoxin cardiotoxin (CTX) into gastrocnemius. The differences in intramuscular inflammation were compared between the wild and knock-out groups on 0, 4, 7 and 10 d after CTX injection by observing exudation of mononuclear phagocytes, macrophages, M1 type macrophages, CD4 +T cells and helpers T cells (Th1, 2&17). Two newborn C57BL/6 wild mice and 2 SM TGF- βr2-/- knock-out mice were selected to culture primary myoblasts in vitro which were divided into 2 groups: an interferon group subjected to interferon simulation and a control group subjected to addition of an equal amount of solvent. The differences in expression of IL-6, IL-10, MCP-1, MIP-1α, H-2K b, H2-Ea, Toll-like receptor (TLR)3 and TLR7 were compared between the interferon and control groups, as well as between the wild and knock-out groups. Results:On 4&7 d after CTX injection, the ratios of mononuclear/macrophage (75.73%±3.62%, 45.27%± 2.32%), macrophages (38.67%±2.76%, 24.87%±2.19%), M1 macrophages (43.21%±0.11%, 30.43%±2.19%), CD4 +T cells (20.13%±1.62%, 5.67%±0.32%) in the muscle tissue from the knock-out mice were significantly higher than those from the wild mice (58.52%±2.43%, 29.21%±2.45%; 20.63%±2.32%, 16.23%±1.25%; 24.98%±0.35%, 14.23%±1.69%; 10.70%±0.43%, 2.50%±0.45%), with a majority of Th1&Th17 ( P<0.05). In vitro results showed that the levels of IL-6, MCP-1, MIP-1α, H-2K b, H2-Ea and TLR3 were significantly upregulated in the interferon group compared with the control group and that such upregulation in the nock-out mice was more significant than in the wild mice ( P<0.05). Conclusions:Endogenous TGF- β signal activation plays a role in the functional recovery after muscle trauma, because it is involved in the regulation of immune behavior of muscle fibers, thus affecting intramuscular inflammation and muscle regeneration.

In order to construct recombinant adenovirus vector expressing Suppressor of cytokine signaling 3 (SOCS3) and obtain infectious adenoviral particles, SOCS3 gene was amplified from plasmid pcDNA3-SOCS3 and subcloned into the adenovirus shuttle plasmid pAdTrack-CMV. After sequence confirmation, the recombinant shuttle plasmid pAdTrack-CMV-SOCS3 was linearized by Pme I, and then transformed into BJ5183 competent cell, the recombinant plasmid pAd-SOCS3 was obtained by homologous recombination between pAdTrack-CMV-SOCS3 and the adenoviral backbone plasmid pAdEasy-1 in BJ5183. The pAd-SOCS3 was linearized by Pac I and transfected into HEK293 cells via liposome. The recombinant adenovirus was packaged and amplified in HEK293 cells. After purifying, virus titer was determined by tissue culture infectious dose 50 (TCID50). Using the recombinant adenoviruses to infect porcine primary adipocytes, the expression of green fluorescent protein (GFP) was observed by fluorescent microscopy, and SOCS3 gene was identified by RT-PCR and Western blotting. Restriction enzyme and PCR analysis demonstrated that the recombinant adenovirus vector was constructed correctly, and the virus titer reached 1.2x10(9) PFU/mL. The result of RT-PCR and Western blotting showed that SOCS3 mRNA and protein expression was remarkably increased in porcine primary adipocytes infected with recombinant adenovirus. In conclusion, this study successfully constructed the recombinant adenovirus containing SOCS3 gene, and can be helpful for further research on the function of SOCS3.
Adenoviridae ; genetics ; metabolism ; Adipocytes ; metabolism ; Animals ; Cloning, Molecular ; Green Fluorescent Proteins ; biosynthesis ; genetics ; HEK293 Cells ; Humans ; RNA, Messenger ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Suppressor of Cytokine Signaling 3 Protein ; Suppressor of Cytokine Signaling Proteins ; biosynthesis ; genetics ; Swine ; Transfection

Swine is an ideal model for diabetes studies. Insulin and insulin resistance are closely related with diabetes. To investigate the effect of SOCS-3 in insulin resistance, porcine primary adipocyte was treated with insulin (100 nmol/L) and dexamethasone (300 nmol/L) to induce insulin resistance. The simi-quantitative PCR results suggested that insulin increased GLUT4, PPARgamma and SOCS-3 gene expression in primary culture porcine adipocytes and no change of OB gene expression. Under insulin resistance conditions, SOCS-3 and OB gene expression were up-regulated, whereas GLUT4 and PPARgamma gene expression were down-regulated in primary porcine adipocytes. The overexpression of PPARgamma gene resulted in the increase of GLUT4 expression by insulin. Different expression levels of SOCS-3 determined the inhibitory effects of insulin signaling. Induction of insulin resistance by dexamethasone was not only due to inhibition of glucose transportation, but also repression of insulin signaling. SOCS-3 might be a potential gene to block the insulin resistance.
Adipocytes ; cytology ; metabolism ; Animals ; Cells, Cultured ; Dexamethasone ; pharmacology ; Glucose Transporter Type 4 ; biosynthesis ; genetics ; Insulin ; pharmacology ; Insulin Resistance ; Leptin ; biosynthesis ; genetics ; PPAR gamma ; biosynthesis ; genetics ; Suppressor of Cytokine Signaling 3 Protein ; Suppressor of Cytokine Signaling Proteins ; biosynthesis ; genetics ; Swine

BACKGROUND:The safety and efficacy of simultaneous bilateral total knee replacement or selective unilateral total knee arthroplasty in patients with severe osteoarthritis of the knees are stil controversial. OBJECTIVE:To compare safety and clinical efficacy of patients with osteoarthritis knees after simultaneous bilateral total knee replacement or selective unilateral total knee replacement. METHODS:Total y 60 cases with severe osteoarthritis of the knees (90 knees) undergoing total knee replacement were divided into unilateral total knee replacement group (n=30, 30 knees), and the simultaneous bilateral total knee replacement group (n=30, 60 knees). RESULTS AND CONCLUSION:There was no significant difference in the incidence of other complications such as infection, mortality, pulmonary embolism in patients of both groups (P>0.05). The incidence of cardiovascular complications, postoperative blood loss and blood transfusion were higher in the bilateral knee group than in the unilateral knee group (P<0.05). During fol ow-up at 1 year after replacement, no significant differences in range of motion, muscle strength of quadriceps and hospital for special surgery knee score were detected in patients of both groups (P>0.05). However, Visual Analogue Scale scores were significantly lower in the bilateral knee group than in the unilateral group (P<0.05). These data indicated that the risk of cardiovascular complications was high in patients receiving bilateral total knee replacement. Patients with severe cardiovascular disease should avoid simultaneous bilateral total knee arthroplasty.

With the development of our economy and the progress of science and technology,the comfort medicine of children in the perioperative period is gradually promoted.Comfort medicine not only eliminates the physical pain of children,but also avoids the permanent psychological trauma caused by oper-ation and anesthesia.The incidence of preoperative anxiety in children is higher than that in adults,which may have adverse effects on perioperative period and prognosis of children.The characteristics of parents of different children,such as parental anxiety level,parental education level,parental socioeconomic back-ground,family composition,ethnicity,language,and other factors will affect the preoperative anxiety level of children.Intervention measures such as parents'participation in guiding children's preparation for anesthe-sia,management of children's fasting and drinking,and presence at the induction of anesthesia can alleviate children's preoperative anxiety level.For children with disabilities,communication between parents and medical staff in the perioperative period and intervention management of children is important.Compared with the traditional preoperative preparation program,the internet-based family preoperative preparation plat-form provides a comprehensive,effective and personalized information guidance for the families of children,ensuring the comfortable perioperative medical treatment of children.

Objective: To explore the correlation between blood pressure and urinary phthalandione, MMP, MEP, MnBP, MiBP, PAEs. Methods: Three schools were selected from Shenzhen, China for the present study. A total of 765 firstgrade students of Han ethnicity were recruited voluntarily from the selected schools during September 2016 to June 2017. They were divided into normal blood pressure (BP) group (lower than P90 group) and high BP group (BP≥P90). Linear and Logistic regression models were used to analyze the relationships between blood pressure and urine phthalate metabolite levels. Results: Urinary MMP and MnBP in students of high BP group were significantly higher than that of students in normal BP group(t=13.12, 3.97, P<0.05). Linear regression models showed that Z score increased when MMP and MnBP levels increased(P<0.05). Logistic regression model suggested that the risk of high BP increased with the increment of MMP level adjusting creatinine, sex, age and BMI(OR=1.47, P<0.05). There was no statistical significance in the differences after adjusting many factors including family income and education level of parents(P>0.05). Conclusion Urinary phthalate metabolite levels are positively associated with blood pressure in first-grade children.