DNA extraction is basis and important step in molecular biology technical. DNA was extracted from peripheral blood by some technique as: Perchlorate sodium, Acetate amonium. QIA amp minikit (Micro technique). All three methods obeyed these steps: destruction of red blood cells, destruction of white blood cells, destruction of protein. Among the techniques mentioned above, perchlorate sodium DNA extraction is the best technique, although it needs only a very small amount of blood (200 microlit) but technique’s price is high. The results indicated: DNA extraction by Perchlorate sodium was the most appropriate with the designed standard. Now, this technique is daily used at Bio-Medical Centre Laboratory, Hanoi Medical University.
DNA ; blood ; Methods

Early diagnosis of sepsis helps make effective clinical decisions and improve the survival rate of patients with severe infection. However, the timely and accurate diagnosis of sepsis is still a great challenge in clinic. In order to settle the very problem, the scientists in the world have made a lot of exploration and research in the field of rapid molecular identification of pathogens. Nowadays, the nucleic acid detection of sepsis is mainly composed of 3 types of methodological strategies, either based on positive blood culture, single colonies, or directly on blood specimens. This paper presents a comprehensive overview of advances in the research of early detection of bacterial nucleic acid as molecular diagnosis of sepsis.
DNA, Bacterial ; blood ; Humans ; Sepsis ; blood ; diagnosis

To evaluate the effects of Dexamethasone(Dex), Platelet derived growth factor-BB(PDGF) and combination of Dex and PDGF(DP) on the growth and differentiation of MC3T3-E1 cells, Dex(10(-7) M) and PDGF(10 ng/ml) in experimental group were added to the cells at the days 5, 10, 15, 20, 25 and examined for cell proliferation activities, DNA synthesis activities, ALP activities and bone nodule formation. The results were as follows : 1.In Dex group, cell proliferation, DNA synthesis and ALP activities were lower until 15 days when compared to the control group. Bone nodules formation were shown at 10 days. 2.In PDGF group, cell proliferation and DNA synthesis activities were higher until 15 days and ALP activities were lower when compared to the control and Dex groups. Bone nodules formation were shown at 20 days. 3.In DP group, cell proliferation and DNA synthesis activities of PDGF were suppressed by Dex and synergistic effects of combination of Dex and PDGF on ALP activities were shown at days 5 when compared to control and Dex groups. Bone nodules formation activities of Dex were suppressed by PDGF.
Blood Platelets ; Cell Proliferation ; Dexamethasone* ; DNA

Objective To evaluate the effects of DNA examination of trace bloodstain samples from the scene collected with Trace Biological Evidence Collection kit. Methods Venous blood was made into bloodstains on the ground. The trace bloodstain samples were collected with Trace Biological Evidence Collection kit and common methods, respectively. DNA examination of trace bloodstain samples （50 from each group） was conducted on the constant temperature shaker for 2, 24, 48, 72, and 96 h, respectively, and the examination results of every group were compared. Results When the trace bloodstain samples were placed on the constant temperature shaker for 24, 48, 72, and 96 h, the DNA detection rates in the group which used Trace Biological Evidence Collection kit （100.00%, 100.00%, 100.00%, 96.00%） were significantly higher than those in the group using common methods （62.00%, 26.00%, 10.00%, 0）, the differences had statistical significance （P<0.05）. When the trace bloodstain samples were placed on the constant temperature shaker for 2 h, the differences of DNA detection rates between the two groups had no statistical significance （ P>0.05）. Conclusion The Trace Biological Evidence Collection kit can effectively improve DNA detection rate and extend detection time limit for trace bloodstain samples from the scene that have been stored for a relatively long time.
Blood Stains ; DNA ; Forensic Medicine ; Temperature

A major problem for the early diagnosis of prostate cancer is the lack of clinically useful tests for screening a preclinical and asymptomic population without resort to invasive diagnostic procedures. Recent studies have demonstrated the possibility to detect genetic alterations in plasma or serum DNA from patients with prostate cancer or other cancers. Quantification and molecular event are associated with advanced stages and circulating tumor cells. These results indicate a new approach to the early diagnosis and monitoring of prostate cancer by non-invasive screening procedures based on the analysis of genetic changes in plasma.
DNA ; blood ; Humans ; Male ; Prostatic Neoplasms ; blood ; diagnosis

BACKGROUNDRecent studies suggest that circulating DNA may be a potential tumor marker for lung cancer, but most of these studies are conducted between healthy controls and lung cancer patients, with few or no benign lung disease patients included. The objective of this study was to evaluate the performance of plasma DNA quantification in discriminating lung cancer from the healthy and benign lung disease.

METHODSPlasma DNA was extracted with a QIAamp DNA Blood Midi kit and quantified by a PicoGreen dsDNA quantitation kit in 44 healthy individuals, 36 benign lung disease patients and 67 lung cancer patients. Discrimination power was evaluated by the receiver operating characteristic curve.

RESULTSPlasma DNA values were significantly increased in lung cancer patients, especially in those with metastases, and in benign lung disease patients compared with that in the healthy individuals (P < 0.001, respectively). The values in lung cancer patients were significantly increased compared with that in the benign lung disease patients (P < 0.001). The area under the curve was 0.96 [95% confidence interval (CI) 0.92 - 0.99] for the healthy versus lung cancer, 0.73 (95% CI 0.64 - 0.83) for lung cancer versus benign lung disease, and 0.86 (95% CI 0.80 - 0.91) for lung cancer versus the healthy and benign lung disease.

CONCLUSIONSPlasma DNA quantification has a strong power to discriminate lung cancer from the healthy and from the healthy and benign lung disease, less power to discriminate lung cancer from benign lung disease. Plasma DNA quantification may be useful as a screening tool for lung cancer.

To explore the application of fetal DNA in maternal plasma for noninvasive prenatal diagnosis, the DNA template was extracted by hydroxybenzene-chloroform from 44 maternal (7-41 weeks) plasma. The Fetus-derived Y sequence DYZ-1 gene (149bp) was chosen to be amplified by PCR. The fragment was identified in all the plasma of male bearing pregnant women with the diagnostic accordance rate being 100.00%. Two of the 22 female bearing pregnant women had false positive results. Among the 44 pregnant women, the diagnostic accordance rate was 88.89% at early pregnant stage, 100.00% at medium pregnant stage, and 96.55% at late stage respectively. The final accuracy of 95.45% was obtained in all cases. It was concluded that by means of hydroxybenzene-chloroform extraction the authors of this article promoted the concentration and purity of the DNA template, and diagnosed more accurately. The results showed that free fetal DNA in the maternal plasma could be regarded as the gene resource for noninvasive prenatal diagnosis.
DNA/*blood ; DNA Primers ; Fetus ; Maternal-Fetal Exchange ; Pregnancy/*blood ; Prenatal Diagnosis/*methods ; *Sex Determination (Genetics)

Biomarkers, Tumor ; blood ; DNA, Neoplasm ; blood ; Humans ; MicroRNAs ; blood ; Molecular Diagnostic Techniques ; Neoplasms ; blood ; diagnosis ; Nucleic Acids ; blood

OBJECTIVES: To evaluate the effects of different pretreatment methods and preservation time on RNA quality of peripheral blood samples, and to optimize the preservation method of peripheral blood samples. METHODS: Eight pretreatment methods were used to preprocess the peripheral blood from 3 healthy unrelated individuals and the treated samples were stored at -80 ℃. Total RNA of samples was extracted using Quick-RNA^TM Miniprep Plus kit. DNA/RNA Shield^TM was added to peripheral blood and total RNA was extracted after preservation at -80 ℃ for 0, 5, 10, 15, 30 and 60 days, respectively. The concentration, purity and integrity of RNA were determined. Statistical analyses were performed by SPSS 22.0 software to compare the differences in RNA yield, purity and integrity among the eight pretreatment methods. RESULTS: In terms of purity, leukocyte pretreated with RNAlater^TM and directly cryopreservation peripheral blood showed the worst purity. The other six methods showed better purity. In terms of yield, blood cells with DNA/RNA Shield^TM came out with the highest yield, followed by peripheral blood with DNA/RNA Shield^TM. In terms of integrity, peripheral blood preserved in PAXgene Blood RNA tube method had the best integrity. Except for peripheral blood pretreated with DNA/RNA Shield^TM and blood cells pretreated with DNA/RNA shield^TM, the other five methods had statistical differences when compared to the method by keeping peripheral blood in PAXgene Blood RNA tube. The purity of RNA stored at six-time gradients ranged from 1.815 to 1.952. With the increase of storage time, RNA yield decreased from 4.516 ng to 1.039 ng, and RNA integrity decreased from 8.533 to 7.150. CONCLUSIONS According to the results of total RNA's yield, purity and integrity, peripheral blood pretreated with DNA/RNA Shield^TM was the best pretreatment method. After the pretreatment, samples can be preserved for up to 60 days in low temperature.
Blood Specimen Collection/methods* ; Cryopreservation ; DNA/analysis* ; Humans ; RNA

(-)-Epigallocatechin gallate (EGCG), a catechin polyphenol component, is the main ingredient of green tea extract. Although the anti-carcinogenic and cancer inhibitory effects of EGCG have been widely reported, its genotoxicity is not clear and seldom reported. In this study, we examined the effects of EGCG on DNA strand breaks in the isolated lymphocytes and whole blood lymphocytes obtained from two smoking subjects and a nonsmoking healthy subject using a single-cell gel electrophoresis (SCG) assay. The results showed that after 2 hrs of treating the isolated lymphocytes from the smokers, EGCG induced a significant increase in DNA strand breaks at concentrations from 2.5 × 10-5 M to 2.0 × 10-4 M, while after 2 hrs of treating the whole blood obtained from the same smokers, EGCG suppressed the DNA strand breaks in the lymphocytes at concentrations of 1.0 × 10-4 M and 2.0 × 10-4 M. A similar suppressive result was also shown in the whole blood lymphocytes from the nonsmoker at nearly the same concentrations, while at concentrations of 1.0 × 10-3 M or 2.0 × 10-3 M, EGCG induced a significant increase in DNA strand breaks in the whole blood lymphocytes from the nonsmoker. This result suggests that EGCG is not only inhibitory against DNA strand breaks in whole blood, but also genotoxic to the isolated or whole blood lymphocytes at high concentrations. Thus, more research is needed to comprehensively assess the effects of EGCG on genetic materials.
Lymphocytes ; Upper case emm ; In Blood ; DNA strand break ; seconds