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

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

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 ; *Blood Specimen Collection ; Edetic Acid/pharmacology ; Hemoglobins/analysis ; Hemolysis/drug effects ; Humans

BACKGROUNDCD4 count is used to determine antiretroviral therapy (ART) eligibility. In China, flow cytometers are mostly located in urban areas with limited access by patients residing in remote areas. In an attempt to address this issue, we conducted a study to validate the performance of Alere PIMA point-of-care CD4 analyzer.

METHODSVenous and finger-prick blood specimens were collected from HIV-positive participants from two voluntary counseling and testing sites in Yunnan Province. Both venous and finger-prick blood specimens were tested with the PIMA analyzer. Venous blood specimens tested with the Becton Dickinson FACSCalibur were used as a reference.

RESULTSVenous specimens from 396 and finger-prick specimens from 387 persons were available for analysis. CD4 counts by PIMA correlated well with those from FACSCalibur with an R2 of 0.91 for venous blood and 0.81 for finger-prick blood. Compared to FACSCalibur, the PIMA analyzer yielded lower counts with a mean bias of - 47.0 cells/μl (limit of agreement, [LOA]: -204-110 cells/μl) for venous blood and -71.0 cells/μl (LOA: -295-153 cells/μl) for finger-prick blood. For a CD4 threshold of 350 cells/μl, the positive predictive value (PPV) of PIMA was 84.2% and 75.7% and the negative predictive value (NPV) was 97.6% and 95.8% for venous and finger-prick blood, respectively. For an ART threshold of 500 cells/μl, the corresponding PPV was 90.3% and 84.0% and NPV was 94.3% and 93.4%, respectively.

CONCLUSIONSCD4 counting using venous blood with PIMA analyzers is a feasible alternative to a large flow cytometer to determine ART eligibility.

Adolescent ; Adult ; Aged ; Biological Assay ; methods ; Blood Specimen Collection ; CD4 Lymphocyte Count ; methods ; Child ; China ; Female ; HIV Infections ; diagnosis ; Humans ; Male ; Middle Aged ; Sensitivity and Specificity ; Young Adult

OBJECTIVES: Excess sodium intake has been linked to obesity and obesity-related indices. However, the scientific evidence for this association is inadequate. The purpose of this study was to investigate the association between urinary sodium excretion and obesity-related indices among Korean adults. METHODS: A convenience sample of 120 subjects (60 obese and 60 non-obese subjects) were recruited applying frequency matching for sex and age between two groups. Sodium intake level was assessed through 24-hour urine collection. Obesity-related metabolic risk factors, including fasting blood lipid indices, subcutaneous and visceral fat through computed tomography (CT), insulin resistance indices, blood pressure and liver enzymes were measured in all subjects. These obesity-related metabolic risk factors were compared between obese and non-obese group according to sodium excretion levels (<110 mEq/day, 110~180 mEq/day, >180 mEq/day). RESULTS: After adjusting for age, gender, health behaviors (smoking, exercise, drinking), and energy intake, several obesity-related metabolic risk factors, including abdominal circumference, body fat percentage, subcutaneous and visceral fat, triglyceride, and systolic blood pressure were found to be significantly deteriorated as the sodium excretion level increases. In addition, multivariate adjusted-odds ratios of abdominal obesity, high blood triglyceride, and high blood pressure were found significantly higher in the highest sodium excretion group compared to the lowest group. The mean number of metabolic syndrome risk factors was also significantly greater in the highest sodium excretion group than in the lowest group. CONCLUSIONS: The current study findings suggested that high sodium intake can affect obesity and metabolic syndrome risk negatively, implying the necessity of future research on low-sodium diet intervention in relation to obesity and related health problems.
Adipose Tissue ; Adult ; Blood Pressure ; Diet, Sodium-Restricted ; Energy Intake ; Fasting ; Health Behavior ; Humans ; Hypertension ; Insulin Resistance ; Intra-Abdominal Fat ; Liver ; Obesity ; Obesity, Abdominal ; Risk Factors* ; Sodium* ; Triglycerides ; Urine Specimen Collection

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 ; Blood Specimen Collection/*instrumentation ; Erythrocytes/*cytology ; Hemolysis ; Humans ; Outpatients

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 ; Blood Pressure* ; Diet ; General Practice ; Humans ; Hypertension ; Primary Health Care* ; Sodium* ; Urine Specimen Collection

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 ; Blood Pressure* ; Diet ; General Practice ; Humans ; Hypertension ; Primary Health Care* ; Sodium* ; Urine Specimen Collection

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 ; Blood Pressure ; Humans ; Hypertension ; Male ; Sodium ; Sodium Chloride ; Taste Threshold* ; Urine Specimen Collection ; Volunteers

OBJECTIVES: This study was conducted to examine blood pressure and other characteristics of a high sodium intake group assessed with 24-hr urine analysis and the dietary factors related to the risk of high sodium intake among Korean adults. METHODS: A cross-sectional study was conducted with adults aged 20-59 years. Subjects who completed 24-hr urine collection (N = 205) were divided into 3 groups (tertile) according to the sodium intake estimated with 24-hour urine analysis. We compared the blood pressure, BMI and dietary related factors of the 3 groups (low, medium, high sodium intake group) with General Linear Model (GLM) and Duncan's multiple range test (p < 0.05). The risk factors related to high sodium intake were assessed with odds ratio (p < 0.05). RESULTS: The sodium intake (mg/day) of the 3 groups were 3359.8 +/- 627.9, 4900.3 +/- 395.1 and 6770.6 +/- 873.9, respectively, corresponding to daily salt intake (g/day) 8.5, 12.4 and 17.2, respectively. High sodium group showed significantly elevated age, BMI and systolic/diastolic blood pressure. Being male gender was associated with significantly increased risk of sodium intake (OR = 1.972; 95%CI: 1.083-3.593). The other factors related to high sodium intake were higher BMI (> or = 25) (OR = 2.619; 95% CI: 1.368-5.015), current alcohol consumption (OR = 1.943; 95%CI: 1.060-3.564), and having salty soybean paste with salt percentage > 14% (OR = 3.99; 95% CI: 1.404-6.841). The dietary attitude related to increased risk of high sodium intake included 'enjoy dried fish and salted mackerel' (p < 0.001) and 'eat all broth of soup, stew or noodle' (p < 0.001). CONCLUSIONS: Because high sodium intake was associated with higher blood pressure, nutrition education should focus on alcohol consumption, emphasis on related dietary factors such as using low salt soybean paste, improvements in the habit of eating dried fish and salted mackerel or eating all broth of soup, stew or noodle.
Adult* ; Alcohol Drinking ; Blood Pressure* ; Cross-Sectional Studies ; Eating ; Education ; Humans ; Linear Models ; Male ; Odds Ratio ; Perciformes ; Risk Factors* ; Sodium* ; Soybeans ; Urine Specimen Collection