1.Effects of Peripheral Blood Different Pretreatment Methods and Preservation Time on RNA Quality.
Jia-Yi ZHANG ; Qian-Nan XU ; Xi-Ling LIU ; Cheng-Tao LI
Journal of Forensic Medicine 2021;37(6):825-831
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-RNATM Miniprep Plus kit. DNA/RNA ShieldTM 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 RNAlaterTM 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 ShieldTM came out with the highest yield, followed by peripheral blood with DNA/RNA ShieldTM. 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 ShieldTM and blood cells pretreated with DNA/RNA shieldTM, 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 ShieldTM was the best pretreatment method. After the pretreatment, samples can be preserved for up to 60 days in low temperature.
Blood Specimen Collection/methods*
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Cryopreservation
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DNA/analysis*
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Humans
;
RNA
2.Comparison of Two New Plastic Tubes (Sekisui INSEPACK and Green Cross Green Vac-Tube) with BD Vacutainer Tubes for 49 Analytes.
The Korean Journal of Laboratory Medicine 2007;27(1):69-75
BACKGROUND: Becton Dickinson (BD) Vacutainer tubes are the most widely used vacuum system for collection of blood in clinical laboratories. We compared the performance of two new tubes, Sekisui INSEPACK tube and Green Cross Green Vac-Tube, with the existing BD Vacutainer tubes for 49 common analytes. METHODS: A total of 20 apparently healthy volunteers were recruited for this study. For rountine chemistry and thyroid function tests, we compared the results of two new vacutainer tubes and BD Vacutainer tubes with those of BD glass tubes at t =0 hr by student paired t test. Hematology and coagulation test results of the two new vacutainer tubes were compared with those of BD Vacutainer tubes at t =0 hr. To study the stability of each analyte, results at t =24 +/- 2 hr, t =72 +/- 2 hr, and t =168 +/- 2 hr were compared with those at t =0 hr for each tube. RESULTS: Although paired t test analysis revealed statistically significant differences between two tested tubes and existing BD Vacutainer tube in several tests (total protein, total bilirubin, alkaline phosphatase, glucose, uric acid, calcium, inorganic phosphorus, direct bilirubin, triglyceride, HDL-cholesterol, LDL-cholesterol, lactate dehydrogenase, triiodothyronine, and thyroxine), these differences were not considered clinically significant. Stability of two new vacuum tubes for each analyte was similar to that of the BD Vacutainer tube. CONCLUSIONS: Sekisui INSEPACK tube and Green Cross Green-Vac Tube showed a satisfactory analytical performance compared with existing BD Vacutainer tubes. We conclude that these two new plastic vacutainer tubes are acceptable for the commonly ordered laboratory tests.
Blood Cells/chemistry
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Blood Proteins/analysis
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Blood Specimen Collection/*instrumentation
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Equipment and Supplies/standards
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Female
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Humans
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Male
3.Effects of Two Placement Ways for Storage of Blood Bag on Biochemical Indexes of Leukodepleted Red Blood Cells.
Rui-Jun ZHANG ; Bing-Zheng DUAN ; Chun-Mei JU ; Su-Qin SUI ; Yan BAI ; Huan CAO
Journal of Experimental Hematology 2016;24(2):607-610
OBJECTIVETo investigate the effects of 2 different ways of storage bag placement on some biochemical indexes of leukodepleted red blood cells (LD-RBC) to as to ensure the efficacy and safety of clinical blood transfusion.
METHODSThe whole blood samples of 20 donors (400 ml/donor) were selected for preparating the LP-RBC, which were divided evenly into 10 bags. The 10 bags were randomly divided into 2 groups; the bags in 1 group were placed uprightly, while the bags in another group were placed horizontally. The bags of 2 groups were stored in the same conditions. One storage bag from each group was taken randomly on day 7, 14, 21, 28, 35 respectively, and then the biochemical indexes of samples were detected and analyzed.
RESULTSThe values of K(+) and LAC on day 14, the value of LDH on day 28 in the uprightly placed group were higher than those in the horizontally placed group (P < 0.05), the value of Na(+) on day 28, and the value of Glu on day 35 in the uprightly placed group were lower than those in horizontally placed group (P < 0.05), but there was no significant difference in Cl(-) level between 2 groups (P > 0.05).
CONCLUSIONThe storage bags placed by different ways during the storage show different influence on some biochemical indexes of LD-RBC in the storage period.
Blood Specimen Collection ; instrumentation ; methods ; Blood Transfusion ; Erythrocytes ; Humans ; Random Allocation
4.Effects of additives in blood collection tubes on testing the alcohol concentration in blood samples.
Journal of Forensic Medicine 2014;30(6):452-455
OBJECTIVE:
To discuss blood collection tubes with different additives and their effects on the testing results of alcohol concentration in blood samples.
METHODS:
Blood samples from 10 volunteers were collected 2 hours after drinking with seven different types of disposable vacuum blood collection tubes, including ordinary tube without anticoagulant, coagulant tube, separating gel-coagulant tube, sodium citrate (1:4) tube, sodium citrate (1:9) tube, sodium citrate (9:1) tube and EDTA-K2 tube. The alcohol concentrations in these blood samples were analyzed by headspace gas chromatography.
RESULTS:
The concentration testing results of the same blood samples in different types of tubes were different from one to another. The sequence was as follows: separating gel-coagulant tube > coagulant tube > ordi- nary tube without anticoagulant > EDTA-K2 tube> sodium citrate (1:9) tube> sodium citrate (1:4) tube, whereas the results of the same blood sample in sodium citrate (1:9) tube and sodium citrate (9:1) tube showed no obvious difference.
CONCLUSION
It is better to collect a suspicious drunk driver's blood sam- ple using a disposable vacuum blood collection tube, with the EDTA-K2 tube being preferred.
Anticoagulants
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Blood Specimen Collection/methods*
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Citrates
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Ethanol/blood*
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Humans
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Sodium Citrate
5.Scoring System for Detecting Spurious Hemolysis in Anticoagulated Blood Specimens.
Gilsung YOO ; Juwon KIM ; Young UH ; Kwang Ro YOON ; Soon Deok PARK ; Kap Jun YOON
Annals of Laboratory Medicine 2015;35(3):341-347
BACKGROUND: The identification of in vitro hemolysis (IVH) using a hematology analyzer is challenging because centrifugation of the specimens cannot be performed for cell counts. In the present study, we aimed to develop a scoring system to help identify the presence of hemolysis in anticoagulated blood specimens. METHODS: Thirty-seven potassium EDTA anticoagulated blood specimens were obtained, and each specimen was divided into 3 aliquots (A, B, and C). Aliquots B and C were mechanically hemolyzed by aspirating 2 and 5 times, respectively, using a 27-gauge needle and then tested; aliquot A was analyzed immediately without any hemolysis. After the cells were counted, aliquots B and C were centrifuged and the supernatants were tested for the hemolytic index and lactate dehydrogenase levels. RESULTS: The 4 hematologic parameters were selected and scored from 0 to 3 as follows:< 34.0, 34.0-36.2, 36.3-38.4, and > or =38.5 for mean cell hemoglobin concentration (MCHC, g/dL); <0.02, 0.02, 0.03, and > or =0.04 for red blood cell ghosts (10(12)/L); <0.13, 0.13-0.38, 0.39-1.30, and > or =1.31 for difference value (g/dL) of measured hemoglobin and calculated hemoglobin; and <0.26, 0.26-0.95, 0.96-3.34, and > or =3.35 for difference value (g/dL) of MCHC and cell hemoglobin concentration mean. The hemolysis score was calculated by adding all the scores from the 4 parameters. At the cutoff hemolysis score of 3, the IVH of aliquots B and C were detected as 64.9% and 91.9%, respectively. CONCLUSIONS: The scoring system might provide effective screening for detecting spurious IVH.
Anticoagulants/*pharmacology
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*Blood Specimen Collection
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Edetic Acid/pharmacology
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Hemoglobins/analysis
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Hemolysis/drug effects
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Humans
6.Comparison of Red Blood Cell Hemolysis Using Plasma and Serum Separation Tubes for Outpatient Specimens.
Dae Hyun KO ; Dahae WON ; Tae Dong JEONG ; Woochang LEE ; Sail CHUN ; Won Ki MIN
Annals of Laboratory Medicine 2015;35(2):194-197
BACKGROUND: To rapidly obtain outpatient results, we use plasma separation tubes (PST) for chemistry analysis. If lactate dehydrogenase measurement is required, serum separation tubes (SST) are used. There has been no evaluation of hemolysis with these tubes. We compared the hemolytic index (HI) obtained by using PST and SST and applied this for choosing appropriate tubes for clinical laboratories. METHODS: The HI of specimens obtained from outpatients visiting Asan Medical Center between July and December 2012 was analyzed. The HI was scored from 0 to 10 by using the Toshiba 200FR (Toshiba Medical Systems Co., Japan). HI was classified by sample tube type, and significant hemolysis was defined as a HI of 2 or more. For significant hemolysis cases, medical records were reviewed to identify the causes. RESULTS: Among 171,519 specimens, significant hemolysis was observed in 0.66% of specimens (0.68% of PST specimens, 0.46% of SST specimens). The mean HI in PST was 0.18 (SD: 0.43) and that in SST was 0.14 (SD: 0.37). The proportion of significant hemolysis was significantly higher in PST than in SST (P=0.001). The cause of significant hemolysis was identified as chemotherapy and prosthetic valve in 48.1% of specimens. Complex sampling errors may have caused significant hemolysis in the remaining 51.9% of specimens. CONCLUSIONS: The incidence of hemolysis was slightly higher for PST than SST, although both were <1%. PST are thought to be more useful than SST in outpatient testing because of rapid turnaround time, greater sample volume, and less risk of random errors due to fibrin strands.
Age Factors
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Blood Specimen Collection/*instrumentation
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Erythrocytes/*cytology
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Hemolysis
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Humans
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Outpatients
7.Plasmodium falciparum Cultivation Using the Petri Dish: Revisiting the Effect of the 'Age' of Erythrocytes and the Interval of Medium Change.
Young A KIM ; Je Eun CHA ; Sun Young AHN ; Seung Ho RYU ; Joon Sup YEOM ; Hyo Il LEE ; Chang Gyun KIM ; Ju Young SEOH ; Jae Won PARK
Journal of Korean Medical Science 2007;22(6):1022-1025
Differences in the characteristics of the culture conditions can influence the multiplication rate of Plasmodium falciparum. The Petri dish method is one of the most popular methods of cultivating this parasite. In many previous studies, ideal culture conditions of the Petri dish method were achieved by using erythrocytes collected from blood that had been stored for at least 2 weeks, with daily changes of the medium. In the present study, we studied the multiplication rate of P. falciparum in cultures containing erythrocytes of various ages together with changing the medium at various intervals of time. Our results strongly suggest that the rate of in vitro multiplication of P. falciparum was higher in freshly collected erythrocytes than in aged erythrocytes regardless of the anticoagulant and that when the parasitemia is lower than 8% with a hematocrit of 5%, the medium change interval can be as long as 48 hr without a great reduction in the rate of multiplication.
Animals
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Blood Specimen Collection
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Cell Aging
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Culture Media
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Erythrocytes/*parasitology
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Plasmodium falciparum/*growth & development
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Time Factors
8.Effect of sodium restriction on blood pressure of unstable or uncontrolled hypertensive patients in primary care.
Willem DE KEYZER ; Katrien TILLEMAN ; Jan AMPE ; Stefaan DE HENAUW ; Inge HUYBRECHTS
Nutrition Research and Practice 2015;9(2):180-185
BACKGROUND/OBJECTIVES: The aims of the present study are: 1) to quantify sodium consumption of patients with unstable or uncontrolled hypertension, 2) to investigate if reduced sodium intake can lower BP in these patients, and 3), to assess the acceptability and feasibility of this approach. SUBJECTS/METHODS: This study included 25 adults (age: 50+ years) with frequently elevated BP or patients with uncontrolled, uncomplicated hypertension despite drug treatment in a general practice setting. BP and salt intake (24h urinary excretion and food records) were measured at baseline and after a sodium reduced diet. RESULTS: Mean (+/- SD) systolic (SBP) over diastolic (DBP) blood pressure (mmHg) at baseline was 150.7 (+/- 9.5)/84.149 (+/- 5.6). Mean urinary sodium excretion was 146 mmol/24h. A reduction of 28 mmol sodium excretion decreased SBP/DBP to 135.5 (+/- 13.0)/82.5 (+/- 12.8) (P < 0.001). After one month of no dietary advice, only in 48%, SBP was still < or =140 mmHg. CONCLUSION: Assessment of sodium intake using food records, 24h urine collections and probing questions to identify use of sodium containing supplements or drugs are essential for tailored advice targeted at sodium intake reduction. The results of the present study indicate that reduced sodium intake can lower BP after 4 weeks in unstable or uncontrolled hypertensive patients.
Adult
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Blood Pressure*
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Diet
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General Practice
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Humans
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Hypertension
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Primary Health Care*
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Sodium*
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Urine Specimen Collection
9.Effect of sodium restriction on blood pressure of unstable or uncontrolled hypertensive patients in primary care.
Willem DE KEYZER ; Katrien TILLEMAN ; Jan AMPE ; Stefaan DE HENAUW ; Inge HUYBRECHTS
Nutrition Research and Practice 2015;9(2):180-185
BACKGROUND/OBJECTIVES: The aims of the present study are: 1) to quantify sodium consumption of patients with unstable or uncontrolled hypertension, 2) to investigate if reduced sodium intake can lower BP in these patients, and 3), to assess the acceptability and feasibility of this approach. SUBJECTS/METHODS: This study included 25 adults (age: 50+ years) with frequently elevated BP or patients with uncontrolled, uncomplicated hypertension despite drug treatment in a general practice setting. BP and salt intake (24h urinary excretion and food records) were measured at baseline and after a sodium reduced diet. RESULTS: Mean (+/- SD) systolic (SBP) over diastolic (DBP) blood pressure (mmHg) at baseline was 150.7 (+/- 9.5)/84.149 (+/- 5.6). Mean urinary sodium excretion was 146 mmol/24h. A reduction of 28 mmol sodium excretion decreased SBP/DBP to 135.5 (+/- 13.0)/82.5 (+/- 12.8) (P < 0.001). After one month of no dietary advice, only in 48%, SBP was still < or =140 mmHg. CONCLUSION: Assessment of sodium intake using food records, 24h urine collections and probing questions to identify use of sodium containing supplements or drugs are essential for tailored advice targeted at sodium intake reduction. The results of the present study indicate that reduced sodium intake can lower BP after 4 weeks in unstable or uncontrolled hypertensive patients.
Adult
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Blood Pressure*
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Diet
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General Practice
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Humans
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Hypertension
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Primary Health Care*
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Sodium*
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Urine Specimen Collection
10.Comparison of Salt Taste Threshold and Salt Intake between Hypertensive and Normotensive Group.
Ho Jin SON ; Jae Jin KO ; Seung Heon SHIN ; Young Soo LEE ; Jong Yeon KIM ; Mee Ra RHYU ; Mi Kyung YE
Korean Journal of Otolaryngology - Head and Neck Surgery 2015;58(10):687-693
BACKGROUND AND OBJECTIVES: Salt-taste threshold can influence salt appetite, and is thought to be another marker of sodium intake. Many studies have found an association between sodium intake and blood pressure. The aim of this study was to compare the salt-taste threshold and salt intake between hypertensive and normotensive groups. SUBJECTS AND METHOD: One hundred twenty volunteers (51 men and 69 women) who did not take antihypertensive medications were evaluated. First, a questionnaire, which included questions regarding demographic information and preference of salty taste, was conducted, and 24-hour ambulatory blood pressure was checked. Then salt taste threshold was measured by assessing the ability of the subjects to discern the taste of salt in graded solutions of saline. Lastly, 24-hour urinary sodium was measured in a 24-hour urine collection. RESULTS: The salt taste threshold and taste preference for salt were slightly higher in hypertensive group. There was slightly higher salt intake measured as 24-hour urinary sodium in the hypertensive group, compared with the normotensive group. However, there were no significant differences in salt taste threshold, preference of salty taste, and salt intake between the normotensive and the hypertensive groups. CONCLUSION: The threshold of salt taste was not related to sodium intake and hypertension status. These results suggest that the development of hypertension depends on the complex interaction of factors such as genes and environmental factors rather than sensory factors like taste threshold and taste preference.
Appetite
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Blood Pressure
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Humans
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Hypertension
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Male
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Sodium
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Sodium Chloride
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Taste Threshold*
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Urine Specimen Collection
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Volunteers