Mononuclear Cell Count Research Articles

Adverse reactions of collection and return infusions of autologous peripheral blood stem cells in children with malignant solid tumors

Objective To study the adverse reactions of collection and return infusions of autologous periphe-ral blood stem cells (APBSC) in children with malignant solid tumors. Methods Thirty-eight cases' clinical data diagnosed as advanced malignant solid tumors in children by pathological examination were collected from September 2005 to November 2011 in Beijing Tongren Hospital (27 male patients and 11 female patients). All cases were given high-dose chemotherapy and APBSC transplantation(APBSCT). According to world health organization(WHO) stan-dard of the adverse reactions of chemotherapy and radiotherapy and the Bearman standard of the adverse reactions of APBSCT, cli-nical data of these cases were analyzed.The adverse reactions of collection and return infusions of APBSC were summarized. Results (1)Mononuclear cell (MNC) count of 38 cases was (5.6±2.2)×108/kg, and CD34+ cell counts was (3.8±2.6)×106/kg.MNC count or CD34+ cell count of 31 cases was conformed to the standards.The coincidence rate was 81.6% (31/38 cases). (2)On (12.3±3.1) days, 38 cases obtained bone marrow reconstruction after return infusions of APBSC.The correlation coefficient of CD34+ cells count and bone marrow reconstruction was rs=-0.634, and that was negatively correlated(P=0.001). (3)Adverse reactions during APBSC collection and return infusions; the most adverse reactions during APBSC collection were arrhythmia, hypotension, gastrointestinal reaction and thrombocytopenia, et al.The main adverse reactions during APBSC return infusions were hemoglobinuria, hypertension and gastrointestinal reaction.The occurring rate of hemoglobinuria was 100%, but urine color gradually returned to normal after they were given the hydration and alkalization.(4) Adverse reactions during bone marrow suppression after return infusions: 38 patients with high-dose chemotherapy on + 3 to + 14 d were presented with toxic side effects those including 19 cases in grade Ⅰ, 11 cases in grade Ⅱ, 5 cases in grade Ⅲ, and 3 cases in grade Ⅳ(but 1 case died in the 3 cases). On + 15 to + 22 d, 37 patients with high-dose chemotherapy, 33 cases showed no adverse reactions, and 2 cases in grade Ⅰ, 2 cases in grade Ⅱ.On + 23 to + 40 d, the adverse reactions of 37 cases with high-dose chemotherapy were corrected. Conclusion The adverse reactions of APBSC were transient and no organ damage was observed, which was safe and reliable. Key words: Solid tumor; Autologous peripheral blood stem cell transplantation; Chemotherapy; Adverse reactions; Child

Type 1 Diabetes Mellitus Donor Mesenchymal Stromal Cells Exhibit Comparable Potency to Healthy Controls In Vitro

The potential for mesenchymal stromal cells (MSCs) as a cell-based therapy in the treatment of immunologic disorders has been well established. Recent studies reported the clinical potential for autologous MSCs as a systemic therapy in the treatment of type I diabetes mellitus (T1D). The current study compared the genotypic and phenotypic profiles of bone marrow-derived MSCs from T1D and healthy donors as autologous (compared with allogeneic) therapy provides distinct advantages, such as reduced risk of immune reaction and transmission of infectious agents. The findings of the current study demonstrate that despite moderate differences in T1D MSCs at the gene level, these cells can be expanded in culture to an extent corresponding to that of MSCs derived from healthy donors. No functional difference in terms of immunosuppressive activity, blood compatibility, or migratory capacity was evident between the groups. The study findings also show that autologous MSC therapy holds promise as a T1D treatment and should be evaluated further in clinical trials.

The Peripheral Blood Mononuclear Cell Count Is Associated With Bone Health in Elderly Men: A Cross-Sectional Population-Based Study.

The peripheral blood mononuclear cell (PBMC) count is a routinely used and meaningful index for infection and blood diseases. PBMCs may be closely related to osteoclasts and include osteoclast precursors; we examined the association between the PBMC count and bone health. This research included 2806 community men aged ≥50 years who underwent full health examinations from October 2007 through December 2011 in four medical centers. The PBMC count was significantly high among subjects with “at least osteopenia” compared with controls. In analysis of covariance adjusted for potential confounders, the bone mineral density (BMD) value and T-score had a significant decreasing trend across the quartiles of PBMC count. In univariate analysis, the PBMC count had a strong association with “at least osteopenia” (odds ratio [OR] = 2.520, 95% confidence interval [CI]: 1.397–4.547). After adjustment for confounding factors (multivariate analysis) from Model 1 to 4, PBMC count remained as an independent risk factor for “at least osteopenia” (OR = 2.481, 95% CI: 1.176–5.236). Moreover, after adjusting for all confounding variables, participants had a significantly high OR in the body mass index (BMI) <25 group (OR = 2.798, CI: 1.122–6.973; P = 0.027) and systolic blood pressure (SBP) <140 group (OR = 2.519, CI: 1.059–5.993; P = 0.037). In conclusion, the PBMC count is significantly associated with bone loss in elderly men and the exact mechanism requires further clarification.

Decision-Tree Algorithm Optimize Hematopoietic Progenitor Cell-Based Prediction in Peripheral Blood Stem Cell Mobilization

Background and ObjectivesEnumeration of hematopoietic progenitor cells (HPC) using an automated hematology analyzer provides rapid, inexpensive, and less technically dependent prediction of peripheral blood stem cell (PBSC) mobilization. This study aimed to incorporate HPC enumeration along with other predictors for optimizing a successful harvest.Materials and MethodsBetween 2007 and 2012, 189 consecutive patients who proceeded to PBSC harvesting with a preharvest HPC ≥ 20 x 106 /L were recruited. A failed PBSC mobilization was defined as < 2 x 106 CD34+ cells/kg. Variables predicting a successful harvest identified by multivariate logistic regression and correlation analysis were subjected to classification and regression tree (CART) analysis.ResultsA total of 154 (81.5%) patients successfully achieved mobilization of CD34+ cells (median 8.18 x 106 CD34+ cells/kg). Five independent host predictors including age ≥ 60, a diagnosis of solid tumor, prior chemotherapy cycles ≥ 5, prior radiotherapy, and mobilization with G-CSF alone or high-dose cyclophosphamide, as well as laboratory markers including HPC and mononuclear cell (MNC) counts, were used for CART analysis. The number of host predictors with a cutoff at two, HPC cutoff at 28 x 106/L and MNC cutoff at 3.5 x 109 /L were best discriminative for successful prediction. In the decision tree algorithm, patients predicted as good mobilizers (0 to 2 risk factors) had a higher success rate (150/169, 88.8%) than that (4/20, 20.0%) of those predicted as poor mobilizers (3-5 risk factors). Moreover, patients predicted as good mobilizers and further with a HPC enumeration ≥ 28 x 106/L had a high probability of achieving successful mobilization (138/148, 93.2%).ConclusionOur CART algorithm incorporating host predictors, HPC enumeration and MNC count may improve prediction and thus increase the success of PBSC mobilization. Further prospective validation is necessary. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

The Effect of High LET Space Radiation on the Human Hematopoietic System: Preliminary Results from the First In Vivo Model

Background: The effects of spaceflight on human physiology are complex and represent a sum of many factors, the main two of which are microgravity and space radiation. The latter is composed of high-energy protons and HZE particles, along with secondary radiation produced in shielding and tissue. One major concern is short- and long-term radiation-induced injury to the hematopoietic system, one of the most radiosensitive tissues in the human body. Previous studies demonstrated functional and/or numerical changes in T and /or B-cells of crewmembers of STS-41B and STS-41D and altered myelopoiesis during STS-63 and STS-69. Although these changes reversed post return, they persisted in space and were more recently documented during a six month mission to the International Space Station. The main goal of NASA's space radiation research program is to enable human space exploration within acceptable risks and to reduce uncertainties regarding risk projections. We present the first ground based in vivo study evaluating the effects of high- and low-LET radiation on the human immune system by using a reconstituted human hematopoietic system in a humanized mouse model. We measure post-radiation depletion and repopulation kinetics, tumor formation and DNA damage.Methods: NSG mice were irradiated with 2Gy of gamma rays and injected 20 hours later with 2x105 CD34+cells isolated from cord blood. After a 12 week engraftment period, peripheral blood flow cytometry with anti-human and anti-mouse antibodies (human and mouse CD45, human CD3 and human CD19) was performed to confirm engraftment (>50% of human cells in moue blood, Figure 1). We irradiated the humanized-NSG mice (hu-NSG) with 0.2 or 0.4 Gy of 350 MeV/n28 Si-ions, which are established components of LET radiation. We measured changes in peripheral blood mononuclear cell counts (huCD45+PBMC) 7 and 30 days post irradiation to assess early effects and recovery kinetics. Multicolor flow cytometric immunophenotyping (FCI) analysis of tissue and blood samples was performed according to standard cell surface staining protocols. Conventional cytogenetic analysis was performed 5 months post radiation on enriched fractions of human HSC and progenitor cells, isolated from marrow of irradiated mice. We performed secondary transplants to transmit the genetic changes imparted by irradiation in the first animal to the secondary host system to allow further monitoring. In addition whole exome and transcriptome sequencing is underway on stem cell and progenitor (lineage negative) pools on a subset of mice.Results: To date 120 well engrafted NSG mice were irradiated with 0.2 or 0.4 Gy of 350 MeV/n28 Si-ions. No decrease versus a mild decrease of huCD45+ PBMCs was observed on day 7 in the 0.2 and 0.4 Gy radiated cohort respectively, with the latter demonstrating full recovery on day 30 compared to normal control mice (Figure 2). No lymphoproliferative or myeloproliferative disorder was noted in 46 irradiated mice using 8 color FCI analyses. Median percentage of peripheral blood mature B-cells, T cells and NK cells was 41%, 36.5% and 9% respectively. Kappa /lambda ratio, CD4/CD8 ratio and NK cell/T cell ratio were 0.9, 3.6, and 0.3 respectively. Bone marrow evaluation demonstrated 0.08% to 0.3% myeloblasts (median: 0.15%) positive for CD34 /CD117/CD33 and HLADR. Moderate monocytosis (CD64+/CD4+/HLA-DR+) was noted, ranging from 39% to 51% (median 47%). Preliminary results of cytogenetic analysis revealed diploid male and female karyotypes without abnormalities in at least 20 metaphases analyzed. Successful secondary transplants were performed in 30 mice. Results of whole exome and transcriptome sequencing are pending.Conclusions: This study shows for first time data on the effects of high -LET space type of radiation on human hematopoietic system in vivo. Preliminary results show that high-LET 350 MeV/n 28 Si radiation has relatively minor effects on peripheral blood cell counts and depletion/recovery kinetics. In addition our preliminary analysis did not demonstrate a leukemogenic effect in the short time period (months) assessed after the irradiation. The study is currently in progress and more data will be available. [Display omitted] [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Isolation of mesenchymal stromal/stem cells from small-volume umbilical cord blood units that do not qualify for the banking system.

The clinical application of mesenchymal stromal/stem cells (MSCs) has been extensively explored. In this study, we examined the availability of freshly donated umbilical cord blood (UCB) units that do not qualify for the Japanese banking system for transplantation because of their small volume as a source of MSCs. Forty-five UCB units were used. The median volume of each UCB unit and number of nucleated cells per unit were 40mL and 5.39×10(8), respectively. MSCs were successfully isolated from 18 of 45 units (40%). The MSC isolation rate was not affected by cell processing method or the interval between delivery and cell processing. The volume of the UCB unit and the mononuclear cell count were predictive factors of the MSC isolation rate. MSCs were effectively isolated by selecting UCB units with a volume of ≥54mL and containing ≥1.28×10(8) mononuclear cells, yielding a MSC isolation rate of >70%. UCB-derived MSCs were similar to bone marrow-derived MSCs in terms of their morphology, surface marker expression, and differentiation potential, apart from adipogenesis. Our data indicate that UCB units that are currently discarded due to inadequate volume should be reconsidered as a source of MSCs using the well-established UCB banking system.

Effects of elevated parameters of subclinical ketosis on the immune system of dairy cows: in vivo and in vitro results

Using an established model in which subclinical ketosis is induced, the response of differential blood counts and levels of various haematological variables, including the inflammatory marker haptoglobin (Hp), were tested over the last six weeks of parturition until the 56th day post-partum in cows with lower or higher body condition scores (LBC and HBC, respectively; n = 9/group). Animals in the HBC group evidenced subclinical ketosis whereas LBC animals were metabolically healthy. For in vitro examination with ß-hydroxybutyrate (BHB) as a further stimulus, peripheral blood mononuclear cell (PBMC) counts of cows with and without subclinical ketosis (n = 5/group) were observed. Counts of leucocytes, granulocytes and lymphocytes (LY) peaked at day 1 post-partum in HBC cows, with a more marked increase in heifers. In subclinical ketosis LY count increased again, with significantly higher values in the HBC group. The red blood cell (RBC) profile was affected by parity (counts were higher in heifers). Hp showed a positive linear correlation with BHB and non-esterified fatty acids (NEFA; R2 = 0.41). PBMC from cows that were not pre-stressed with subclinical ketosis were more sensitive to increasing levels of BHB in vitro, as evidenced by both their higher proliferative capability and increased release of nitric oxide (NO). In summary, cows with subclinical ketosis showed a heightened immune response compared with metabolically healthy individuals, based on increased LY counts, increasing stimulative properties of PBMC and a relationship between Hp and typically increased values of BHB and NEFA. Concentrations of BHB in vivo during subclinical ketosis did not alter the proliferative capability of bovine PBMC in vitro, which was first significantly decreased at a dosage of 5 mM BHB.

Yield Prediction of Nucleated Cells As Well As CD34+ Progenitors in Allogeneic Bone Marrow Harvests Depends on Differentially Important Biological Donor Variables. A Retrospective Single Center Analysis of 1275 Allogeneic Bone Marrow Collections

Cell requests for allogeneic bone marrow grafts are defined by the amount of total nucleated cells (TNC) according to body weight of the recipient. However, outcome of bone marrow transplantation significantly depends on the amount of infused CD34+ stem and progenitor cells representing around 0.5 to 1.5% of TNC. So far, TNC as well as the CD34+ progenitor yield of marrow harvests cannot be predicted. We retrospectively analyzed donor variables, TNC and progenitor yields of 1275 marrow collections (860 male and 415 female) that have been performed in our collection center - all of those with same experienced staff of physicians. Maximum amount of bone marrow that was allowed to harvest was 20 ml/kg donor weight but limited to a maximum of 1500 ml. The mean TNC count in the final product was analyzed with 16.6 TNC/nl (5.9-46.6) corresponding to 5.6 ± 4.3 TNC/kg body weight of the recipient. The mean CD34+cell concentration in the graft was measured with 111/μl (11-674) corresponding to 4.3 ± 3.9 CD34+cells/kg. We fitted linear regression model for TNC adjusted for harvest volume and recipient weight to identify influential donor variables. Model selection was performed by backward elimination (BE) with the Bayesian information criterion (BIC). Peripheral blood leucocytes of the donor (WBC) before harvest, constitutional donor variables (weight and height), age, platelet count (PLT), smoking status and mononuclear cell count (MNC) before harvest could be identified as predictive donor parameters for TNC-concentration in the bone marrow product – ordered in descending mode by Wald-test statistic. Interestingly, the influence of donor parameters for CD34+ progenitor cell concentration in the marrow was different from TNC. Constitutional parameters of the donor were identified to be most important for CD34+ cell concentration in the marrow product followed by WBC, age and PLT, whereas smoking status and MNC did not influence CD34+ cell harvest. With sex specific centered parameters donor sex was only influential - for both TNC as well as CD34+ cells - through the constitutional parameters weight and height. No significant interactions could be found between harvested volume and the identified donor parameters. Taking also the recipient weight into account we propose a model that allows identification of risk constellations for achieving the designated amount of TNC as well as CD34+ hematopoietic progenitor cells from bone marrow harvests. In addition, this prediction model will enable us to identify additional genetic determinants for collection efficiency in bone marrow.

Equine bone marrow volume reduction, red blood cell depletion, and mononuclear cell recovery using the PrepaCyte-CB processing system.

Volume reduction and RBC depletion of equine bone marrow specimens are necessary processing steps for the immediate therapeutic use of bone marrow (BM)-derived mesenchymal stem cells (MSC), and for MSC expansion in culture. The purpose of the study was to evaluate the ability of the PrepaCyte-CB processing system to reduce volume, deplete RBC, and recover mononuclear cells (MNC) from equine BM specimens. One hundred and twenty mL of heparinized BM were obtained from each of 90 horses. A CBC was performed on the BM pre- and post-PrepaCyte-CB processing. Volume and RBC reduction, and total nucleated cell (TNC) and MNC recoveries were determined. Bone marrow volume was reduced from 120 mL to 21 mL with a median RBC depletion of 90.1% (range, 62.0-96.7%). The median preprocessing total TNC count was 2.2 × 10(9) (range, 0.46-7.9 × 10(9)) and the median postprocessing TNC count was 1.7 × 10(9) (range, 0.3-4.4 × 10(9); P < .0001), with a median recovery of 73.5% (range, 22.4-216.7%). The median preprocessing total MNC count was 0.9 × 10(9) (range, 0.1-4.7 × 10(9)) and median postprocessing total MNC count was 0.8 × 10(9) (range, 0.1-2.7 × 10(9); P = .06), with a median recovery of 83.7% (range, 15.4-413.9%). The PrepaCyte-CB processing system can be used to deplete both volume and RBC, and recover MNC from equine BM specimens. Further studies assessing the viability of MSC and the efficacy of MSC expansion after using the PrepaCyte-CB processing system are warranted.

Haploidentical hematopoietic stem cell transplantation with supplemental umbilical cord blood infusion in treatment of malignant hematological diseases

Objective To evaluate the efficacy of haploidentical hematopoietic stem cell transplantation (HSCT) with supplemental umbilical cord blood (UCB) infusion in treatment of malignant hematological diseases. Method Clinical data of 66 patients with hematological malignancies treated with HSCT in our hospital between January 2010 and May 2013, were retrospectively analyzed. Among them 25 cases received infusion of human UCB before HSCT (experimental group) and other 41 cases had no UCB injection before HSCT (control group). Results There were no differences in age, gender, donor type, disease categories, disease status before transplant between two groups (P>0.05). There was a significant difference in conditioning regimes between two groups (P<0.05), but no clinical implication. The infused mononuclear cell (MNC) count in experimental group was higher than that in control group (9.94±2.88×108/kg vs. 7.80±0.82×108/kg, P=0.00), while there were no difference in infused CD34+ cell count (5.46±3.54×106/kg vs. 3.54±1.60×106/kg, P=0.16). Neutrophil recovery time in experimental group was shorter than that in control group (13.7±2.9 d vs. 16.6±2.9 d, P=0.023). The incidences of grade Ⅲ-Ⅳ acute graft versus host disease (aGVHD, P=0.036), bacterial infection(P=0.001)and fungal infection(P=0.001)and hemorrhagic cystitis (P=0.00)in experimental group were lower than those in control group. There were no significant differences in platelet recovery time(P=0.43), the incidence of grade Ⅰ-Ⅱ aGVHD(P=0.27), implanted syndrome(P=0.24), sinusoidal obstruction syndrome(P=0.57)and viraemia(P=0.31)between two groups. Conclusion HSCT with supplemental infusion of human UCB may alleviate the degree of aGVHD, but the long-term outcome remains to be studied. Key words: Hematopoietic stem cell transplantation; Fetal blood; Hematologic neoplasms

Body fluid cellular analysis using the Sysmex XN‐2000 automatic hematology analyzer: focusing on malignant samples

The majority of previous studies on body fluid (BF) mode of automatic hematology analyzer used nonmalignant BF samples. Here, we evaluated the BF mode on the recently launched Sysmex XN for counting blood cells, especially for malignant samples. A total of 405 BF specimens including 125 malignant samples were analyzed using both the automated method and manual microscopy. In non-cerebrospinal fluids (CSF) samples, there was an agreement between two methods for WBC, RBC, polymorphonuclear, and mononuclear cell counts (R(2) =0.96, 0.94, 0.88, and 0.88, respectively). CSF samples showed slightly poorer correlations than other fluids. Exclusion of malignant samples significantly improved correlations in non-CSF samples, but not in CSF samples. High fluorescence-BF (HF-BF) cells were identified significantly more frequently in malignant samples compared to benign samples (17.8 and 4.15/100 WBC, respectively; P<0.001). Receiver operating characteristic curve analysis demonstrated an HF-BF cell AUC of 0.791 using a cutoff value of 6.9/100 WBC for detecting malignant samples. The BF mode on the Sysmex XN could be an alternative method for the manual counts in the BF analysis with a few drawbacks. However, if a concentration of HF-BF cells is greater than the given threshold, microscopic examination should be subsequently performed.

Synovial fluid differential cell count in wear debris synovitis after total knee replacement.

Determining the cause of synovitis following total knee arthroplasty (TKA) can be challenging. The differential diagnoses include infection, hemarthrosis, instability, crystalline disease, wear debris or idiopathic causes. Wear particle synovitis can mimic periprosthetic infection with symptoms of pain and effusion. Radiographs and physical exam are often inconclusive in differentiating the two. Synovial fluid analysis is routinely used in evaluating periprosthetic infections. We examined the association between synovial white blood cell count and differentials, and polyethylene wear and osteolysis, to see if fluid analysis can aid in establishing the diagnosis of wear particle synovitis. A cell count and differential was obtained from synovial fluid samples from 54 TKAs undergoing revision for aseptic failure. Explanted polyethylene inserts were analyzed for linear and volumetric wear, oxidation (ketone peak height), and damage features. Analysis was performed to assess the relationship between cell counts and polyethylene wear indicators as well as severity of intra-operative and radiographic osteolysis. Total and percent mononuclear (monocyte and lymphocyte) cell counts were found to be elevated in the presence of documented wear debris synovitis and an association was suggested between their levels and maximum ketone levels. The present study implies that the differential cell count of knee fluid can help distinguish wear debris from infection as a source of synovitis following TKA and identifies the value of the mononuclear cell count as a possible tool to assess abnormal wear rates of the polyethylene insert. Further research into identifying the exact role of monocytes in the wear debris synovitis and osteolytic pathways is warranted. Level II, diagnostic study.

The composite of bone marrow concentrate and PRP as an alternative to autologous bone grafting.

One possible alternative to the application of autologous bone grafts represents the use of autologous bone marrow concentrate (BMC). The purpose of our study was to evaluate the potency of autologous platelet-rich plasma (PRP) in combination with BMC. In 32 mini-pigs a metaphyseal critical-size defect was surgically created at the proximal tibia. The animals were allocated to four treatment groups of eight animals each (1. BMC+CPG group, 2. BMC+CPG+PRP group, 3. autograft group, 4. CPG group). In the BMC+CPG group the defect was filled with autologous BMC in combination with calcium phosphate granules (CPG), whereas in the BMC+CPG+PRP group the defect was filled with the composite of autologous BMC, CPG and autologous PRP. In the autograft group the defect was filled with autologous cancellous graft, whereas in the CPG group the defect was filled with CPG solely. After 6 weeks radiological and histomorphometrical analysis showed significantly more new bone formation in the BMC+CPG+PRP group compared to the BMC+CPG group and the CPG group. There were no significant differences between the BMC+CPG+PRP group and the autograft group. In the PRP platelets were enriched significantly about 4.7-fold compared to native blood. In BMC the count of mononuclear cells increased significantly (3.5-fold) compared to the bone marrow aspirate. This study demonstrates that the composite of BMC+CPG+PRP leads to a significantly higher bone regeneration of critical-size defects at the proximal tibia in mini-pigs than the use of BMC+CPG without PRP. Furthermore, within the limits of the present study the composite BMC+CPG+PRP represents a comparable alternative to autologous bone grafting.

Influence of age on the proliferation of adipose-derived stem cells in vitro

Objective In order to study the age associated changes,the characteristics of adipose derived stem cells (ADSCs) from 26 to 45 years old healthy volunteers were investigated.Methods Subcutaneous adipose tissue was obtained by tumescent liposuction from healthy women donors.ADSCs were isolated with collagenase digestion.We compared cells from 4 different age groups in 24 female patients ranged from 26 to 45 years old.Characteristics of growth and proliferation capacities of ADSCs were accessed in vitro.ADSCs were characterized by MTT and cell cycle detection methods.Results There was no significant difference of the mononuclear cells count between the four age groups while the first passage time of second and third group were shorter than that of third and fourth group,respectively (P＜0.05).A significant increase in quiescence of S phase was observed in ADSCs from aged donors.ADSCs of third age group showed stronger capability than the fourth one (P＜0.05).The ADSCs MTT activity of fourth group was significantly lower than the previous 3 groups (P＜0.05).Conclusions Proliferation capacities of ADSCs in the group of older than 40 years decrease significantly compared with the younger groups.The results have significantly implicated that proliferation capacities of ADSCs are reduced with aging. Key words: Subcutaneous fat; Adipose-derived stem cells (ADSCs); Age factor; Cell proliferation; Cultured

Influence of weight and structure on biological behavior of polypropylene mesh prostheses placed at the esophageal hiatus.

Experimental knowledge about mesh behavior at the esophageal hiatus is rare, but such information is essential in order to find a safe and effective method of mesh reinforcement. This study aimed to investigate the influence of mesh structure on the biological behavior of polypropylene prostheses placed at the hiatus. Twenty-four pigs in three groups of eight underwent implantation of heavyweight small-porous (HW-SP), heavyweight large-porous (HW-LP), or lightweight large-porous (LW-LP) circular polypropylene mesh at the hiatus. Eight weeks later, the meshes were explanted. Macroscopic analysis was performed evaluating mesh deformation, adhesions, and position relative to the hiatal margin. Histological analysis comprised evaluation of foreign body reaction and tissue integration by mononuclear cell count and immunostaining of Ki-67, collagen type I, and collagen type III. No mesh-related complications occurred. Mesh shrinkage was observed within all groups and was the lowest for HW-LP, higher for HW-SP, and highest for LW-LP (13.8% versus 19.5% versus 25.5%; P<.001). The adhesion score was highest for HW-SP, lower for HW-LP, and lowest for LW-LP (11.0 versus 8.0 versus 6.0; P<.001). The collagen type I/III ratio was higher for HW-SP compared with HW-LP and LW-LP (3.1 versus 2.2 versus 1.8; P=.014). Heavyweight polypropylene meshes may be advantageous for application at the hiatus. They provide a solid fixation of the esophagogastric junction by adhesions, which may contribute to a reduction of hernia recurrence. In heavyweight meshes, the large-porous structure is associated with superior form stability, and small-porous meshes are superior with regard to solidity of tissue integration, which may prevent mesh migration.

Effects of paroxetine on cutaneous wound healing in healthy and diabetic rats.

The aim of this study was to evaluate the histologic effects of acute paroxetine administration on wound healing in healthy and streptozotocin-induced diabetic rats. This study has a randomized controlled experimental design. Healthy (n = 32) and diabetic (n = 32) rats were further divided into 2 groups of saline or paroxetine administration. Sixty-four male Sprague-Dawley rats were used in this study. Paroxetine was injected intraperitoneally every day. Full-thickness excision wounds were created with a 4-mm dermal punch on the back of all rats. The healing wound area was removed with a 6-mm dermal punch at postwounding days 1, 3, 7, and 14. Polymorphonuclear leukocyte, mononuclear inflammatory cell, fibroblast, and blood vessel counts and epithelialization were evaluated under light microscope. There was no statistically significant difference observed in the polymorphonuclear leukocyte, mononuclear inflammatory cell, and blood vessel counts in the healthy and diabetic rats with and without paroxetine administration. The number of fibroblasts was significantly higher at postwounding day 14 of the paroxetine-administered healthy rats compared with the saline-administered healthy rats (P = .04). However, the number of fibroblasts did not show any difference by paroxetine administration in the diabetic rats. There was no statistically significant difference in epithelialization regarding all the postwounding days, but complete epithelialization was observed in all rats on postwounding day 14 in the healthy and paroxetine-administered group. Short-term paroxetine administration may enhance cutaneous wound healing by increasing the number of fibroblasts and causing better epithelialization over time in healthy rats but not in diabetic rats.

Oral doxycycline for Lyme neuroborreliosis with symptoms of encephalitis, myelitis, vasculitis or intracranial hypertension

The treatment recommendation for Lyme neuroborreliosis with central nervous system (CNS) symptoms is intravenous ceftriaxone, according to current American and European guidelines. For Lyme neuroborreliosis with peripheral nervous system (PNS) symptoms, treatment with intravenous ceftriaxone and oral doxycycline is considered equally effective. The purpose of this study was to evaluate the efficacy of oral doxycycline in the treatment of Lyme neuroborreliosis with CNS symptoms. Patients with Lyme neuroborreliosis who had undergone cerebrospinal fluid (CSF) sampling before and after treatment at the Department of Infectious Diseases, Sahlgrenska University Hospital, during the period 1990-2012, were included in this retrospective study. The CSF mononuclear cell count was used as a surrogate marker of treatment outcome. Comparisons of CSF mononuclear cell counts were made between patients with CNS symptoms and patients with PNS symptoms before and after treatment with oral doxycycline. Twenty-six patients classified as having CNS symptoms and 115 patients classified as having PNS symptoms were included. The decline in CSF mononuclear cell counts did not differ significantly between the two groups of patients. All patients with CNS disease showed a marked clinical improvement after treatment, even though 62% had remaining symptoms at the end of follow-up. Treatment with oral doxycycline resulted in a similar decrease in CSF mononuclear cell counts in patients with Lyme neuroborreliosis with CNS symptoms compared with patients with Lyme neuroborreliosis with PNS symptoms. The results indicate that oral doxycycline is an effective treatment for Lyme neuroborreliosis irrespective of the severity of symptoms.

Reduced Intensity Conditioning, Combined Transplantation of Haploidentical Hematopoietic Stem Cells and Mesenchymal Stem Cells in Patients with Severe Aplastic Anemia

We examined if transplantation of combined haploidentical hematopoietic stem cells (HSC) and mesenchymal stem cells (MSC) affected graft failure and graft-versus-host disease (GVHD) in patients with severe aplastic anemia (SAA). Patients with SAA-I (N = 17) received haploidentical HSCT plus MSC infusion. Stem cell grafts used a combination of granulocyte colony-stimulating factor (G-CSF)-primed bone marrow and G-CSF-mobilized peripheral blood stem cells of haploidentical donors and the culture-expanded third-party donor-derived umbilical cord MSCs (UC-MSCs), respectively. Reduced intensity conditioning consisted of fludarabine (30 mg/m2·d)+cyclosphamide (500 mg/m2·d)+anti-human thymocyte IgG. Transplant recipients also received cyclosporin A, mycophenolatemofetil, and CD25 monoclonal antibody. A total of 16 patients achieved hematopoietic reconstitution. The median mononuclear cell and CD34 count was 9.3×108/kg and 4.5×106/kg. Median time to ANC was >0.5×109/L and PLT count >20×109/L were 12 and 14 days, respectively. Grade III-IV acute GVHD was seen in 23.5% of the cases, while moderate and severe chronic GVHD were seen in 14.2% of the cases. The 3-month and 6-month survival rates for all patients were 88.2% and 76.5%, respectively; mean survival time was 56.5 months. Combined transplantation of haploidentical HSCs and MSCs on SAA without an HLA-identical sibling donor was safe, effectively reduced the incidence of severe GVHD, and improved patient survival.

Circulating Endothelial Progenitor Cells as Markers for Severity of Ischemic Chronic Heart Failure

IntroductionDespite a high potential of endothelial progenitor cells (EPCs) for diagnostic purposes, the EPC role in developing ischemic chronic heart failure (CHF) has not been determined obviously.The objective of this study was to assess the counts of CD45+CD34+, CD45−CD34+, CD14+CD309+, and CD14+CD309+Tie2+ phenotyped circulating EPCs of various subpopulations in patients with ischemic CHF. Methods and ResultsThe study involved 153 patients (86 male), aged 48–62 years, with angiographically proven coronary artery disease (CAD) and 25 healthy volunteers. CHF was diagnosed in 109 patients (71.2%). Mononuclear cell populations were phenotyped by flow cytofluorimetry. Cardiovascular risk factors, such as type 2 diabetes mellitus, hyperlipidemia, arterial hypertension, and adherence to smoking, may have a negative effect on circulating EPC counts in CAD patients regardless of the presence of CHF. The depletion of the CD14+CD309+- and CD14+CD309+Tie2+-phenotyped circulating EPC counts is associated with the severity of left ventricular dysfunction, whereas the CD45+CD34+- and CD45−CD34+-mononuclear cell counts are more representative of the severity of atherosclerotic coronary artery lesions. ConclusionThe authors found that New York Heart Association functional class of CHF, left ventricular ejection fraction <42%, the N-terminal pro–B-type natriuretic peptide level >554 pg/mL, and Е/Еm ratio >15 U had the highest predictive value for the depletion of the EPC count in CAD patients.

Clinical value of hematologic test in predicting tumor response to neoadjuvant chemotherapy with esophageal squamous cell carcinoma

BackgroundTo investigate the relationship between hematologic test results and the predictive effect of regression of esophageal cancer after neoadjuvant chemotherapy (NACT), we analyzed pre-NACT hematologic data and their relationship to tumor regression.MethodsThirty-eight consecutive patients with locally advanced squamous cell esophageal carcinoma who had undergone two cycles of paclitaxel/carboplatin NACT were enrolled. On the day prior to the first cycle of chemotherapy, hematologic tests, including routine blood test and biochemical examinations, were recorded. All patients were confirmed to have no history of hepatitis. Surgical resection was performed when clinical restaging showed effective regression. Histopathological examination was routinely performed to evaluate the postoperative effects of chemotherapy.ResultsAfter two cycles of NACT, tumor imaging evaluation showed that 27 of the 38 patients had CR and PR, including 25 patients who underwent radical esophagectomies. Six patients had stable disease and five patients had progressive disease. According to the hematologic test results before NACT, patients with higher white blood cell counts, lymphocyte percentages, mononuclear cell counts, neutrophilic granulocyte counts, and eosinophilic granulocyte counts and lower alanine aminotransferase (ALT) level had a significantly greater opportunity for an effective response.ConclusionBasal host immunologic function and hepatic function are associated with tumor response to NACT in patients with esophageal cancer. These parameters may have a certain predictive efficacy on NACT for esophageal squamous cell carcinoma.