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

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

No abstract available.
Adosterol/diagnostic use ; Adrenal Cortex/radiography/*radionuclide imaging ; Adrenal Cortex Neoplasms/pathology/radiography/*radionuclide imaging/surgery ; Adrenalectomy/methods ; Adrenocortical Adenoma/pathology/radiography/*radionuclide imaging/surgery ; Aged ; Aldosterone/blood ; Blood Specimen Collection/methods ; Humans ; Hyperaldosteronism/radiography/*radionuclide imaging ; Male ; Radiopharmaceuticals/diagnostic use ; Tomography, X-Ray Computed

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 ; Blood Specimen Collection/methods* ; Citrates ; Ethanol/blood* ; Humans ; Sodium Citrate

OBJECTIVETo compare the effect of different types and concentrations of sweet solutions on neonatal pain during heel lance procedure.

METHODTotally 560 full term neonates (male 295, female 265) were randomized into 7 groups:placebo group (plain water), 10% glucose, 25% glucose, 50% glucose, 12% sucrose, 24% sucrose and 30% sucrose groups.In each group, 2 ml corresponding oral solutions were administered through a syringe by dripping into the neonate's mouth 2 minute before heel lance. The procedure process was recorded by videos, from which to collect heart rate, oxygen saturation and pain score 1 min before puncture, 3, 5 and 10 min after puncture.

RESULTThe average heart rate increase 3, 5 and 10 min after procedure in the 25% and 50% glucose groups, 12% and 24% and 30% sucrose groups was significantly lower than those in the placebo group (P < 0.01 or 0.05). The average heart rate increase 3 min after procedure in the sucrose group was lower than that in the glucose group (P < 0.01).Neonates who received 30% sucrose has a significantly lower average heart rate increase than those who received 12% and 24% sucrose 3 min after heel lance (both P < 0.05) . The average oxygen saturation decrease 3, 5, 10 min after procedure was significantly lower than those in the placebo group (P < 0.01). The average oxygen saturation decrease 3 min after procedure in the sucrose groups was significantly lower than that in the glucose groups (P < 0.01). The average pain score 3, 5, 10 min after procedure was significantly lower than those in the placebo group (P < 0.01). The average pain score 3 min after procedure in the sucrose groups was significantly lower than that in the glucose groups (P < 0.01).

CONCLUSIONOral administration of sweet solutions is an effective way to relieve neonatal pain on procedure, and sucrose has a better pain relief action than glucose, moreover, 30% sucrose provides better effect in control of heart rate increase 3 min after heel lance, but the best concentration of sucrose for pain relief needs further study.

Administration, Oral ; Analgesics ; administration & dosage ; therapeutic use ; Blood Specimen Collection ; adverse effects ; methods ; Facial Expression ; Female ; Glucose ; administration & dosage ; therapeutic use ; Heart Rate ; Heel ; Humans ; Infant, Newborn ; Male ; Oxygen ; blood ; Pain ; physiopathology ; prevention & control ; Pain Measurement ; Sucrose ; administration & dosage ; therapeutic use ; Sweetening Agents ; administration & dosage ; therapeutic use

PURPOSE: This study was done to compare the rates of hemolysis and repeated sampling in blood samples obtained by a syringe needle versus a vacuum tube needle. METHODS: A randomized, prospective study was used to evaluate the differences between the two blood sampling methods. The study group consisted of patients seen in the emergency department (ED) for blood sampling to determine electrolyte level. ED patients were randomly assigned to either the syringe group or the vacuum tube group. All blood samples were collected by experienced ED nurses and hemolysis was determined by experienced laboratory technologists. Data were analyzed using Fisher's exact test and binary logistic regression. RESULTS: One hundred forty-five valid samples were collected (74 in the syringe group versus 71 in the vacuum tube group). 5 of 74 (6.8%) blood samples in the syringe group and 8 of 71 (11.3%) in the vacuum tube group hemolyzed. Repeated blood sampling occurred for 2 of 74 (2.7%) and 3 of 71 (4.2%) in each group respectively. There were no significant differences in rates of hemolysis and repeated sampling between two groups (B=1.97, p=.204; B=2.36, p=.345). CONCLUSION: Venipuncture with syringe needles can be recommended for ED nurses to obtain blood samples.
Adult ; Aged ; Blood Specimen Collection/instrumentation/*methods ; Emergency Service, Hospital ; Female ; *Hemolysis ; Humans ; Logistic Models ; Male ; Middle Aged ; Phlebotomy ; Prospective Studies ; Questionnaires ; Syringes

BACKGROUNDBlood culture contamination is a significant adverse event. The aim of this project was to evaluate the efficacy of a strict blood collection procedure in reducing the blood culture contamination rate.

METHODSA prospectively controlled study was performed in two different medical areas in Peking Union Medical College Hospital (PUMCH) for 16 months (from May 2006 to September 2007). In test group, a strict blood collection procedure was carried out by trained nurses with the veinpuncture sites were scrupulously disinfected with 2.5% tincture of iodine plus 70% alcohol. In control group, commonly used procedure in PUMCH was performed with 0.45% chlorhexidine acetate plus 0.2% iodine. Blood culture positive results for 4 target organisms (Coagulase-negative staphylococci, Propionibacterium acnes, Corynebacterium species and Bacillus species) were further assessed by physicians from infectious department to determine whether a sample was true positive (pathogen) or false positive (contamination).

RESULTSTotal 9321 blood culture collections were analyzed. The blood culture contamination rate in test group was significantly lower than that in control group (5/3177 (0.16%) vs. 77/6144 (1.25%); χ(2) = 13.382, P < 0.001). The most common contaminant was Coagulase-negative staphylococcus (76.83%). The average cultural time during which contaminated samples became positive was longer than that for true pathogen samples (42.0 hours vs. 13.9 hours, P = 0.041).

CONCLUSIONUsing a strict blood collection procedure can significantly reduce blood culture contamination rate.

Anti-Infective Agents, Local ; pharmacology ; Bacillus ; drug effects ; Blood ; microbiology ; Blood Specimen Collection ; adverse effects ; methods ; Chlorhexidine ; pharmacokinetics ; Corynebacterium ; drug effects ; Disinfection ; methods ; Humans ; Iodine ; pharmacology ; Propionibacterium ; drug effects ; Prospective Studies ; Staphylococcus ; drug effects

The study was aimed to compare the safety of hematopoietic stem cell mobilization and collection in related donors providing bone marrow and peripheral blood stem cells and in unrelated donors providing peripheral blood stem cells only. 100 related donors from September 2005 to August 2006 at Institute of Hematology & People Hospital, Peking University, and 71 unrelated donors from November 2003 to December 2007 in Data Bank of Chinese Hematopoietic Stem Cell Donor Beijing Management Center, were observed in process of bone marrow and peripheral blood stem cell mobilization, collection, and follow-up. The change of hematologic parameters (white blood cell count, platelet count and hemoglobin level) and the side effects were recorded and evaluated on months 1, 3 and 6 as well as annually after PBSC donation. During follow-up, long-term side effects and life quality were investigated by questionnaires. The results showed that the total MNC count of bone marrow and PBSC from related donors was 6.70 (4.11 - 12.23) x 10⁸/kg, and the total CD34(+) cell count was 3.40 (1.61 - 13.57) x 10⁶/kg; the total MNC count of PBSC from unrelated donors was 6.69 (3.35 - 11.48) x 10⁸/kg, and the total CD34(+) cell count was 3.50 (1.15 - 11.60) x 10⁶/kg. The main side effect of mobilization was bone pain, reported in 47.0% of the related donors and in 43.7% of unrelated ones, the main side effect of collection was paresthesia, reported in 25.0% of the related donors and in 29.6% of unrelated ones, there was no significant difference on side effects between related and unrelated donors during mobilization and collection of hematopoietic stem cells, all donors could endure these side effects, and no donor discontinued G-CSF administration because of side effects. After collection, the hemoglobin level of related donors was lower than that of unrelated donors [(125.8 ± 20.2) g/L vs (143.2 ± 20.1) g/L] (p < 0.05) because of bone marrow and peripheral blood collection, and the platelet count of unrelated donors were lower than that of related donors [(126.2 ± 57.2) x 10⁹/L vs (162.4 ± 72.9) x 10⁹/L] (p < 0.05) because of more than two times of collection. There was no significant difference on hematologic parameters between two groups during long-term follow-up, and the majority of the donors reported were in good or very good health. It is concluded that the donation proved from related and unrelated donors is safe to mobilize hematopoietic stem cells for allogeneic transplantation. Long-term monitoring of healthy PBSC donors remains important to guarantee the safety standards of bone marrow and peripheral blood stem cell mobilization and collection, including comprehensive medical examination before mobilization and collection, careful manipulation during collection, long term follow up after collection and so on.
Adolescent ; Adult ; Blood Donors ; Blood Specimen Collection ; adverse effects ; methods ; Female ; Hematopoietic Stem Cell Mobilization ; methods ; Hematopoietic Stem Cell Transplantation ; adverse effects ; methods ; Humans ; Male ; Middle Aged ; Transplantation, Homologous ; Young Adult

The study was aimed to explore the effects of recombination human granulocyte colony-stimulating factor (rhG-CSF) on Th17 cells in donors' peripheral blood (GPB) and bone marrow grafts (GBM). 25 healthy donors were injected subcutaneously with rhG-CSF 5 µg/(kg·d) for 5 consecutive days. GBM and GPB were harvested after injection on day 4 and 5 respectively. Some of these donors' steady-state bone marrow (SSBM) and steady-state peripheral blood (SSPB) were harvested before rhG-CSF injection. The changes of IL-17 secreted by T cells in donor BM and PB before and after mobilization were detected by flow cytometry. The results showed that the ability to secrete IL-17 from CD4(+) T cells and CD8(+) T cells in GBM was significantly lower than those in SSBM (GBM vs SSBM Th17/CD4(+) T, 0.74% ± 0.27% vs 1.78% ± 1.19%, p < 0.05; Tc17/CD8(+)T, 0.19% ± 0.16% vs 0.36% ± 0.37%, p < 0.05), changes in peripheral blood and bone marrow were same (GPB vs SSPB Th17/CD4(+) T, 1.82% ± 0.91% vs 3.26% ± 1.89%, p < 0.01; Tc17/CD8(+) T, 0.21% ± 0.17% vs 0.44% ± 0.28%, p < 0.01). The ratios of Th17/CD4(+) T and Tc17/CD8(+) T in GPB were higher than in GBM (p < 0.05, p < 0.01). It is concluded that rhG-CSF in vivo can inhibit the generation of Th17 cells both in bone marrow and peripheral blood grafts, and it may be partial reason for GPB/GBM mixed transplantation without increasing the GVHD incidence.
Adolescent ; Adult ; Blood Donors ; Blood Specimen Collection ; Child ; Female ; Granulocyte Colony-Stimulating Factor ; pharmacology ; Hematopoietic Stem Cell Mobilization ; methods ; Humans ; Male ; Middle Aged ; Recombinant Proteins ; Th17 Cells ; drug effects ; Young Adult