20?10 9/L) was observed (mean 41 days, range 12 to 118 days). There w as the association between the number of CD34 +CD44 + cells infusion and time to neutrophic recovery (?= -0.657 , P

The objective of this research was to explore whether the number of CD34(+)CD38(+) cells infused affects hematopoietic reconstitution after cord blood transplantation. The number of CD34(+)CD38(+) cells in cord blood was analysed with flow cytometry after freezethawing. The body weight and time for neutrophil and platelet recovery were measured in 20 children with acute leukemia. The results showed that the median number of CD34(+)CD38(+) cells infused was 29.47 (9.85 - 325.71) x 10(4)/kg. A median time for neutrophil recovery (> 5 x 10(8)/L) in 20 patients was 18.5 (11 - 32) days, and time for platlet recovery (> 2 x 10(10)/L) in 19 of 20 patients was 45 (12 - 118) days. The number of CD34(+)CD38(+) cells infused correlated with time to neutrophil and platelet recovery (r = -0.577, P < 0.01 and r = 0.503, P < 0.05, respectively). In conclusion, the number of CD34(+)CD38(+) cells infused is correlated with the time for hematologic recovery.
ADP-ribosyl Cyclase ; analysis ; ADP-ribosyl Cyclase 1 ; Adolescent ; Antigens, CD ; analysis ; Antigens, CD34 ; analysis ; Child ; Child, Preschool ; Fetal Blood ; cytology ; transplantation ; Hematopoiesis ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Leukemia, Myeloid, Acute ; blood ; therapy ; Membrane Glycoproteins ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; blood ; therapy

OBJECTIVESeveral studies have shown that L-selectin on CD34-positive cells play a role in hematopoietic reconstitution after peripheral blood stem cell transplantation and allograft bone marrow transplantation. This study sought to investigate whether the numbers of CD(34)(+)CD(62L)(+) cells infused affect the engraftment of hematopoietic stem cells (HSC) and the time to neutrophil and platelet recovery after unrelated umbilical cord blood transplantation for the treatment of childhood acute leukemia.

METHODSTwenty-three children with acute leukemia who received unrelated umbilical cord blood transplantation of mostly mismatched HLA locus were included in this study. Flow cytometry was used to count the numbers of CD(34)(+)CD(62L)(+) cells after freezing-thawing by labelling the cells with anti-CD(34) and anti-CD62L. The patients' clinical data including body weight, engraftment of the HSC, times to neutrophil and platelet recovery were evaluated.

RESULTSTwenty-one patients who received CD(34)(+)CD(62L)(+) cell infusion at a number ranging from 1.37 x 10(5)/kg to 2.68 x 10(6)/kg (median, 3.567 x 10(5)/kg) had successful engraftment of the unrelated umbilical HSC. The numbers of CD(34)(+)CD(62L)(+) cells infused were statistically different between patients who had successful engraftment of the umbilical HSC and those who had not (P < 0.05). The engraftment occurred more commonly in patients who received > 1.3 x 10(5) CD(34)(+)CD(62L)(+) cells/kg. The time of neutrophil recovery (> 500/ microl) ranged from 11 days to 32 days (median, 17.5 days). The data of the time to platelet recovery (> 2 x 10(5)/ microl) were obtained in 18 patients, and it ranged from 12 days to 118 days (median, 14 days). There seemed to be a tendency of correlation between the numbers of CD(34)(+)CD(62L)(+) cells infused and time to platelet recovery (gamma = -0.324, 0.05 < P < 0.1), whereas the numbers of CD(34)(+)CD(62L)(+) cells infused correlated with the time to platelet recovery (gamma = -0.470, P < 0.05).

CONCLUSIONThis study suggests that the numbers of CD(34)(+)CD(62L)(+) cells infused might be involved in the engraftment of HSC and hematologic reconstitution after umbilical cord blood transplantation.

Acute Disease ; Adolescent ; Antigens, CD34 ; blood ; Blood Platelets ; metabolism ; Child ; Child, Preschool ; Cord Blood Stem Cell Transplantation ; methods ; Female ; Humans ; Infant ; Infusions, Intravenous ; L-Selectin ; blood ; Leukemia ; immunology ; therapy ; Male ; Neutrophils ; metabolism ; Treatment Outcome

The study was aimed to establish a protocol of isolating and culturing adult mesenchymal stem cells (MSC) from human bone marrow aspirate and identify them by surface antigen analysis and committed differentiation in order to provide an experimental foundation for achieving a therapeutic benefit in applying MSC in hematopoietic stem cell transplantation. MSCs were obtained from fresh human bone marrow aspirate by gradient centrifugation with Percoll (1.073 g/ml) and anchoring culture in L-DMEM with 10% fetal bovine serum by a full medium exchange every 3 days. The MSC surface antigens, including CD34, CD45, CD73, CD105, CD166, were analyzed on FACScan flow cytometer. Under culture in conditioned medium for osteogenesis (the hormone cocktail containing 0.1 micromol/L dexamethasone, 10 mmol/L glycerol-2-phosphate and 50 micromol/L ascorbic acid) and adipogenesis (the cocktail containing 1 micromol/L dexamethasone, 5 mg/L insulin, 0.5 mmol/L 1-methyl-3-isobutylxanthine and 60 micromol/L indomethacin), MSCs committedly differentiated into osteoblasts and adipocytes. The differentiated mesenchymal stem cells were identified by morphological observation and immunohistochemical staining. The results showed that by gradient centrifugation and adhesion culture, MSCs could be isolated and culture-expanded from human bone marrow aspirate. These cells were uniformly negative for CD34, CD45 and positive for CD73, CD105 and CD166. The osteogenic differentiated cells were positive for alkaline phosphatase (ALP) and the adipogenic differentiated cells displayed accumulation of lipid vacuoles, as detected by oil red O. It is concluded that MSC can be isolated and expand-cultured from adult human bone marrow aspirate and committedly differentiate into osteoblasts and adipocytes. MSC primary identification can be accomplished by flow cytometry and induced differentiation. The set of methods in current experiment shows somewhat practical value for basic research and clinical application.
5'-Nucleotidase ; metabolism ; Antigens, CD ; metabolism ; Bone Marrow Cells ; cytology ; Cell Adhesion Molecules, Neuronal ; metabolism ; Cell Culture Techniques ; Cell Differentiation ; physiology ; Cell Separation ; methods ; Endoglin ; Fetal Proteins ; metabolism ; Humans ; Mesenchymal Stromal Cells ; cytology ; Receptors, Cell Surface ; metabolism

Neuronal firing is crucial to the information processing in the nervous system. In order to make a further study of bifurcation scenarios, experiments were performed on neural pacemakers formed at the injured site of rat sciatic nerve subjected to chronic ligatures. We chose the conductance of voltage-dependent potassium ion channels as conditional parameter, and the extracellular calcium concentration as bifurcation parameter, to give a demonstration of how the firing pattern of neural pacemaker responses to dual parameter adjusting. Among 28 preparations observed, 21 were insensitive to dual parameter adjusting since no change of bifurcation scenario structure was detected. On the contrary, the residual 7 preparations showed dramatic bifurcation scenario shifting corresponding to different dual parameter configuration. Briefly, when concentration of 4-aminopyridine (4-AP), a voltage-dependent potassium ion channels blocker, was kept at different level and extracellular Ca2+ concentration was decreased gradually, different bifurcation scenarios of firing patterns were exhibited in an identical neural pacemaker. The two-parameter bifurcation scenarios of experimental neural pacemaker with different parameter configuration were also different. The results show that neural firing pattern is different when the parameter configuration is different, and the bifurcation scenario is a fundamental framework to identify the transitions between firing patterns.
4-Aminopyridine ; pharmacology ; Action Potentials ; physiology ; Animals ; Calcium ; metabolism ; Male ; Neurons ; physiology ; Periodicity ; Potassium Channels ; physiology ; Rats ; Rats, Sprague-Dawley ; Sciatic Nerve ; injuries ; physiopathology

OBJECTIVE: To explore the change rules of peak area ratio of STR loci to Amelogenin (AMEL) locus (STR/AMEL), a sex-determining gene in DNA degradation, and to evaluate the application of STR/AMEL value in the estimation of DNA degradation degree. METHODS: DNA was extracted from iliopsoas, and the variations of STR/AMEL value (Penta E/AMEL, Penta D/AMEL, FGA/AMEL) were analyzed after the artificial degradation was made by DNase I, and the changes of these three ratios of the iliopsoas naturally degraded in an outdoor environment were also analyzed. The regression curves were analyzed using the periods of DNA degradation and outside the body as the independent variable (x) and the STR/AMEL value as the dependent variable (y) and three curve equations under two conditions were established. RESULTS: Both under the conditions of artificial and natural degradation, STR/AMEL value had a negative relationship with the degradation time. The relationship between STR/AMEL and degradation time can be well simulated by the cubic function. R2 was over 0.99 under controlled degradation condition and over 0.86 under natural degradation condition. CONCLUSION The STR/AMEL value (Penta E/AMEL, Penta D/AMEL, FGA/AMEL) is negatively related with the DNA degradation degree, which follows mathematical regression models strictly, and it might be applied to evaluate the DNA degradation degree.
Amelogenin/genetics* ; DNA Damage/genetics* ; DNA Primers ; Humans ; Microsatellite Repeats ; Regression Analysis ; Time Factors

Umbilical cord blood (UCB) is an alternative source of hematopoietic stem cells for transplantation with success being associated with the total nucleated cell (TNC) count, CD34(+) cells and colony-forming unit-granulocyte-macrophage (CFU-GM) content infused. This study was purposed to clarify the impact of maternal and neonatal factors on hematopoietic potential of UCB product. UCB samples were screened, processed, tested and cryopreserved according to the Standard Operation Procedure (SOP) of Guangzhou cord blood bank (GZCBB). Relationship of hematopoietic cell parameters with maternal and neonatal characteristics for 4615 UCB units was analyzed retrospectively. The results showed that both collected volume (Mean ± SD: 95.23 ± 22.42 ml; Median: 91.85 ml) and initial TNC [Mean ± SD: (1.34 ± 0.49) × 10(9); Median: 1.25 × 10(9)] correlated well with postprocessed TNC [Mean ± SD: (1.21 ± 0.42) × 10(9); Median: 1.14 × 10(9); p < 0.001], CD34(+)count [Mean ± SD: (5.14 ± 4.55) × 10(6); Median: 4.08 × 10(6); p < 0.001] and CFU-GM content [Mean ± SD: (9.72 ± 8.66) × 10(5); Median: 7.53 × 10(5); p < 0.001]. As for donor factors, only infant birth weight correlated strongly with volume collected and all hematopoietic cell parameters (p < 0.001). UCB samples from bigger babies had higher collected volume, TNC, CD34(+) count and CFU-GM content (p < 0.001). Mother's age had no correlation with all the above parameters. Gestational age correlated positively with initial/postprocessed TNC (p < 0.001) and negatively with CD34(+) count (p = 0.04), but no relation with collected volume and CFU-GM content. Cesarean section produced superior volume (Mean ± SD: 97.05 ± 22.23 ml vs 92.53 ± 22.43 ml; Median: 94.08 ml vs 88.82 ml; p < 0.001), but inferior cell count than vaginal delivery (p < 0.001). Male infants had more initial volume and CD34(+) count (Mean ± SD: 96.41 ± 22.31 ml vs 93.95 ± 22.47 ml; Median: 93.27 ml vs 90.14 ml; p < 0.001); [Mean ± SD: (5.28 ± 5.04) × 10(6) vs (5.00 ± 3.94) × 10(6); Median: 4.18 × 10(6) vs 3.94 × 10(6); p < = 0.042], but lower initial and postprocessed TNC than female ones [Mean ± SD: (1.31 ± 0.50) × 10(9) vs (1.37 ± 0.47) × 10(9); Median: 1.22 × 10(9) vs 1.28 × 10(9); p < 0.001]; [Mean ± SD: (1.18 ± 0.42) × 10(9) vs (1.24 ± 0.41) × 10(9); Median: 1.10 × 10(9) vs 1.17 × 10(9); p < 0.001], while no significant difference of CFU-GM were found between male and female infants. It is concluded that these data may be helpful to optimize the UCB donor selection and improve cost efficiency of UCB bank resource. The heavier infants after vaginal delivery should be selected and large-volume units with higher TNC should be chosen at first.
Adult ; Birth Weight ; Blood Banks ; methods ; Cord Blood Stem Cell Transplantation ; methods ; Delivery, Obstetric ; Donor Selection ; Female ; Fetal Blood ; cytology ; immunology ; Gestational Age ; Hematopoietic Stem Cells ; Humans ; Infant, Newborn ; Male ; Maternal Age ; Pregnancy ; Young Adult

To study the program of evaluating mothers and infants after 6 months of cord blood donation, from June 1998 to February 2004, all mothers after 6 months of cord blood donation were followed-up by phone calls or letters to report on the health condition. The results showed that when 3 195 mothers were visited by phone calls, 18 mothers declined to answer. 392 letter were send to those who could not be found by phone, 15 of whom wrote back. The average time to talk with each mother was approximately 12 minutes. Follow-up on the baby donors showed two cases with chromosome abnormality, one with hypothyroidism, one with neutropenia, one with albinism and 5 dead with unclear reasons. The cord blood components from all these abnormal donors found were discarded. In conclusion, the programs to evaluate mother and baby after 6 months of cord blood donation seems important in quality control of the components stored in cord blood bank.
Adult ; Blood Banks ; Blood Donors ; China ; Female ; Fetal Blood ; Follow-Up Studies ; Humans ; Infant ; Infant, Newborn ; Quality Assurance, Health Care ; methods ; statistics & numerical data ; Surveys and Questionnaires ; Time Factors

BACKGROUNDThe influences of genomic background are confirmed in more diseases. Immunologic tolerance after intrauterine infection of hepatitis B virus is considered to occur in T cells. Cytokines work effectively in eliminating virus by immune system after hepatitis B virus infection. To explore the relationship between cytokines (tumor necrosis factor-alpha, interferon-gamma, interleukin-4 and interleukin-10), which expressed abnormal quantity in the peripheral blood to intrauterine hepatitis B virus infectious children, gene single nucleotide polymorphism (SNP) and susceptibility to hepatitis B virus intrauterine infection.

METHODSThis is a cross sectional study of molecular clinical epidemiology. The subjects in this study were selected from outpatients of hepatitis B vaccine follow-up special clinics of our hospital in the period. According to intrant criteria, the high risk children of hepatitis B virus (HBV) intrauterine infection were divided into immune failure group (group I); and immune effective group (group II) and non high risk children belonged to the control group. Four gene SNP sites of TNF-alpha -238, IFN-gamma +874, IL-4 -590 and IL-10 -1082 were determined by real-time quantitative fluorescent polymerase chain reaction (PCR).

RESULTSThe significant differences of TNF-alpha -238 A allele frequency were found between group I and group II (chi(2) = 6.797, P < 0.05) and between group I and the control group (chi(2) = 9.513, P < 0.05). No evident differences of TNF-alpha -238 A were found between group II and control group (chi(2) = 0.047, P > 0.05); the significant differences of IFN-gamma +874 A allele frequency were found between group I and group II (chi(2) = 7.238, P < 0.05), and between group I and the control group (chi(2) = 5.199, P < 0.05). No evident differences were found between group II and the control group (chi(2) = 0.602, P > 0.05); the significant differences of IL-4 -590 C/T allele frequency were not found between group I and group II (chi(2) = 0.632, P > 0.05), also group I and the control group (chi(2) = 0.584, P > 0.05), and the group II and the control group (chi(2) = 0.004, P > 0.05) respectively; The significant differences of IL-10 -1082 G allele frequency were found between group II and group I (chi(2) = 10.359, P < 0.001), and between group II and the controls (chi(2) = 35.418, P < 0.001), but the significant differences were not found between group I and the control group (chi(2) = 1.759, P > 0.05).

CONCLUSIONSThis study suggested the possibility that the TNF-alpha -238 A allele and IFN-gamma +874 A allele were associated with HBV intrauterine infection. There was no evident relationship between IL-4 -590 C/T allele SNP and susceptibility to HBV intrauterine infection, but the IL-10 -1082 G allele was associated with preventive efficacy to HBV intrauterine infection.

Cross-Sectional Studies ; Cytokines ; genetics ; Female ; Genetic Predisposition to Disease ; Genotype ; Hepatitis B ; genetics ; transmission ; Humans ; Infant, Newborn ; Infectious Disease Transmission, Vertical ; Interferon-gamma ; genetics ; Interleukin-10 ; genetics ; Interleukin-4 ; genetics ; Male ; Pregnancy ; Tumor Necrosis Factor-alpha ; genetics

AIMTo study the pharmacokinetics and relative bioavailability of probucol inclusion complex capsule.

METHODSFollowing oral administration of a single dose of 250 mg of conventional tablet (formulation A, purchased from the market) and probucol inclusion complex capsule (formulation B, a new formulation for preclinical trial) to each of 6 healthy dogs in a randomized crossover design, the plasma levels of the active drug at different time points were determined by HPLC and the plasma concentration-time profiles of formulation A and B were obtained. The pharmacokinetic parameters as well as relative bioavailability were analyzed.

RESULTSThe concentration-time curves of formulation A and formulation B were found to fit a two-compartment open model. The Tmax values of formulation A and formulation B were (9.3 +/- 2.1) h and (9.3 +/- 2.1) h, the Cmax values were (1.5 +/- 1.0) microgram.mL-1 and (2.3 +/- 0.9) microgram.mL-1 and the AUC0-240 values were (85 +/- 56) microgram.h.mL-1 and (134 +/- 55) microgram.h.mL-1, respectively. The relative bioavailability of formulation B was found to be (198 +/- 90)% compared with formulation A. The results of variance analysis and two one-side t-test showed that there was significant difference between the two formulations in the AUC0-240.

CONCLUSIONThe high bioavailability by the inclusion of formulation B is attributed to the improvement of its water-solubility by the inclusion process and this is supposed to be a key factor for improving drug bioavailability.

Administration, Oral ; Animals ; Anticholesteremic Agents ; administration & dosage ; pharmacokinetics ; Biological Availability ; Capsules ; Dogs ; Female ; Probucol ; administration & dosage ; pharmacokinetics ; Random Allocation ; Tablets