Meleagris gallopavo) and determine the effects of age, sex, and beak size on beak shape variation using geometric morphometrics. Dorsal and right lateral images were taken of 2442 turkeys at 6 and 18.5 weeks of age. Landmarks were digitized in tpsDig in three analyses of the dorsal upper mandible, lateral upper mandible, and lateral lower mandible shape of each turkey at both ages. The coordinate data were then subjected to a principal components analysis (PCA), multivariate regression, and a canonical variates analysis (CVA) with a Procrustes ANOVA in MorphoJ. For the dorsal images, three principal components (PCs) showed beak shape variation ranged from long, narrow, and pointed to short, wide, and blunt upper mandibles at both ages . Three PCs showed the lateral upper mandible shape variation ranged from long, wide beaks with long, curved beak tips to short, narrow beaks with short, pointed beak tips at both ages . Three PCs also explained 97.80% (6 weeks) and 97.11% (18.5 weeks) of the lateral lower mandible shape variation ranging from wide and round to narrow and thin lower mandibles with superior/inferior beak tip shifts. Beak size accounted for varying proportions of the beak shape variation  in the three analyses of each age group. For all the analyses, the CVA showed sexual dimorphism in beak shape (P < 0.0001) with female upper mandibles appearing wider and blunter dorsally with long, curved beak tips laterally. Whereas male turkey upper mandibles had a narrow, pointed dorsal appearance and short, pointed beak tips laterally. Future applications of beak shape variability could have a genetic and welfare value by incorporating beak shape variation to select for specific turkey beak phenotypes as an alternative to beak treatment.The objective of this study was to assess beak shape variation in domestic turkeys ( A significant proportion of mortalities and culls in domestic turkeys show signs of injurious pecking, which suggests that this damaging behaviour contributes to decreased productivity and economic losses in commercial production. Injurious pecking also represents a serious welfare concern for domestic turkeys –3. The eInfrared laser treatment is currently the most common form of beak trimming used in domestic turkeys and it is typically performed at the hatchery on day-old turkeys . Comparea priori assumptions regarding what measurements should be taken [Traditional analyses of beak shape in poultry have used linear measurements of length, depth, and width to describe variation in beak morphology . Howeverbe taken ,33.The objective of this study was to evaluate the phenotypic variation in turkey beak shape using landmark-based geometric morphometrics, and to determine if age, sex, and beak size had an effect on the beak shape variation in domestic turkeys. To get a comprehensive understanding of turkey beak morphology, we examined the dorsal and lateral shape variation of the upper and lower mandibles of domestic turkeys in three analyses at two ages. Determining the amount of phenotypic variation in beak shape within domestic turkeys in this study will then allow for an investigation of the genetic basis of beak shape variation. If beak shape variation shows a high response to selection, the possibility exists for future breeding to select for domestic turkeys with a reduced capacity to cause pecking damage as an alternative to beak treatment.The experimental protocol in this study was approved by the University of Guelph’s Animal Care Committee  in strict accordance with the recommendations outlined by the University of Guelph Animal Care Policy and the Canadian Council for Animal Care .ad libitum feed and chlorinated water from shared feeders and drinkers [Beak morphology data were collected on male and female male-line Hybrid convertor turkeys (n = 2442) with known pedigree information. These turkeys came from two groups hatched two weeks apart in May—June 2014. At one day of age, turkeys were de-snooded then individually marked with numbered and bar-coded yellow plastic, tab end wing bands . The female and male turkeys from each hatch were housed together in a single power-ventilated, close-sided free-run barn and then separated into single sex flocks at 7 weeks of age in individual barns. The turkeys were housed under standard commercial conditions and fed a standard diet of drinkers .Each turkey was photographed at two ages: 6 and 18.5 weeks of age. The age class of 18.5 weeks was the average age of turkeys photographed for the second analysis of beak shape between 17–20 weeks of age. Photographs were taken as TIFF image files using a Canon Powershot G16 camera  on a black wooden L-shaped platform composed of two black boards  with the horizontal board secured to an adjustable camera tripod . Both platform boards included a 5 cm ruler for later scaling. A 5 cm plastic strip was also included on the horizontal board to ensure consistent positioning of the turkeys’ heads in the photographs. For each data collection, two photographs were taken from the dorsal and right lateral view of a turkey’s head. Dorsal photographs were taken with the camera at the top edge of the vertical platform board and captured a complete image of a turkey’s head from the beak tip to the base of the skull . The rige.g., the tip in the upper mandible). In contrast, semilandmarks are points defined by extrinsic criteria and are commonly used to provide more shape information when traditional landmarks are unavailable [Three analyses of beak shape were performed on the dorsal and right lateral images from each turkey at each age. Two analyses of the right lateral image provided a cross section of the beak shape of the upper and lower mandibles separately. A specific set of landmark and semilandmark coordinates (LM) were placed on the beak images in tpsDig version 2.29  in the dvailable ,37. In tvailable ,37.Before the coordinates were applied, the scaling factor for measurement was set in tpsDig using the ruler that was included in the background of each image. The images were rotated (if necessary) to reduce extraneous variation in the placement of the coordinates. Dorsal images were rotated until a straight line could be drawn from LM 1 and a central line between LM 2 and 3 . The latThe semilandmarks were positioned using a standardized grid and placed where the grid lines intersected the outer margins of the mandible being analyzed ,31. For i.e., the multidimensional measurement of standard deviation representing the distance between an individual shape measurement and the consensus shape), morphometric beak data was available for 2429 dorsal, 2099 lateral upper mandible, and 2081 lateral lower mandible images for the six-week old turkeys [Multivariate statistical shape analyses of the turkey beak images were completed using tpsRelw version 1.65  and Morp turkeys . For thek to 2k – 4  [The raw coordinates from each turkey image were first aligned through translation, scaling, and rotation using a generalized least squares Procrustes superimposition algorithm adjusting for sliding semilandmarks in tpsRelw. The aligned shape coordinates were then analyzed in MorphoJ –32. The ndmarks) . The algndmarks) –42. In tndmarks) ,41,43. Tndmarks) .In MorphoJ, a principal components analysis (PCA) was performed using the covariance matrix of the Procrustes shape coordinates to identify the main orthogonal axes of beak shape variation within each dataset as individual principal components . A canonThe dorsal images of upper mandible included morphometric data from 1186 female and 1243 male turkeys at six weeks of age. The PCA of the dorsal Procrustes shape coordinates for the six-week old turkeys concentrated 95.36% of the explained total variation within the first three principal components (PCs). PC1 explained 74.56% of the shape variation in the dorsal shape profile of the upper mandible ranging between long, narrow beaks with pointed tips to short, wide beaks with blunt tips . PC2 expP < 0.0001), which increased slightly when pooled by sex . Along the axis of centroid size, centroid size explained the variation from long, narrow beaks with pointed tips to short, wide beaks with blunt beak tips for the dorsal upper mandible shape of both male and female turkeys. The CVA produced a single variate that explained 100% of the dorsal beak shape variation between male and female turkeys at 6 weeks of age. This variate showed female turkeys had a slightly wider dorsal upper beak outline shown as an outward shift of the semilandmarks (LM 4–13) and a more blunt beak tip (LM 1) than the pointed beaks of male turkeys at this age  ranging from upper mandibles with a long, narrow dorsal beak shape with pointed tips to short, wide beaks with blunt tips . PC2 refP < 0.0001). The larger centroid size of male turkeys  explained the longer and narrower shape of the dorsal upper mandibles with more pointed beak tips . The CVA of the dorsal upper mandible images produced one variate that explained 100% of the shape variation between the sexes at this age . Partitioning the data by sex marginally increased the amount of lateral upper mandible shape variation explained by beak size . Along the axis of centroid size, the lateral upper mandible shape of both male and female turkeys varied from wide beaks with more curved tips to narrow beaks with more pointed tips. The CVA produced one variate that explained 100% of the variance in lateral upper mandible shape between the 1089 female and 1010 male turkeys in this group .Beak size accounted for 13.31% of the total lateral shape variation in the upper mandible for the six-week old turkeys . The variation in the second (14.28%) and third principal components (7.61%) described the slight widening/narrowing of the lateral upper mandible shape along with the superior/inferior shifting of LM 1, which showed the range from beaks with short, pointed tips to upper mandibles with long, curved beak tips  ranging from short, narrow upper mandibles with short, pointed tips to long, wide beaks with long, curved beak tips . Fig 9 iP < 0.0001). Male turkeys had larger centroid sizes, which explained a more superiorly positioned and pointed upper mandible tip .For the 18.5-week old turkeys, beak size only accounted for 0.97% of the total lateral shape variation in the upper mandible, which increased slightly to 1.03% when pooled by sex  with an associated superior/inferior shift in the beak tip LM 1; . In contP < 0.0001). Along the axis of centroid size, the lateral lower mandible shape of both sexes varied slightly from narrow to wide beaks. One canonical variate explained 100% of the variation between the lateral lower mandible beak shape of female and male turkeys at six weeks of age. The lower mandibles of female turkeys were rounder and wider with more inferiorly positioned beak tips than the more thin, narrow beaks with superior positioned beak tips of male turkeys at this age  explained a majority of the shape variation ranging from wide and round to narrow and thin lower mandibles (LM 4–13) with superior/inferior shifts in the position of the beak tip LM 1; . SimilarP < 0.0001). Male turkeys had larger centroid sizes (mean centroid size: 9.03 ± 0.03 mm) that explained the wider shape of the lateral lower mandibles (LM 4–13) with more superiorly shifted beak tips (LM 1). In contrast, female turkeys at this age had smaller centroid sizes (mean centroid size: 7.44 ± 0.03 mm), which explained more narrow lateral lower mandibles (LM 4–13) with more inferiorly positioned beak tips  that have been reported in the lateral upper mandible profiles of other bird species [Analysis with landmark-based geometric morphometrics showed a wide range of phenotypic shape variation in the beaks of these domestic turkeys. For all three beak analyses, the main axes of beak shape variation were relatively consistent between the two ages. The dorsal outline of upper mandible showed a main axis of shape variation from long, narrow, and pointed to short, wide, and blunt beaks at both 6 and 18.5 weeks of age. Similarly, the majority of shape variation in lateral images of the lower mandible at both ages ranged from wide and round to narrow and thin lower mandibles with superior/inferior shifts in the lower beak tips. The lateral upper mandible showed a main axis of shape variation from long, wide upper beaks with long, curved tips to short, narrow beaks with short, pointed tips at both 6 and 18.5 weeks of age. The main shape axis for these turkeys parallels the main patterns of shape variation  might be less constrained by beak size differences. However, more research is needed to substantiate the hypothetical relationships between the size and shape variation in the different beak structures of domestic turkeys.In the three analyses of beak morphology, variation in beak size predicted varying amounts of the beak shape variation in these turkeys. Beak size explained approximately 35–55% of the explained morphological variation in the lower mandibles of the six-week old turkeys and the dorsal upper mandibles of both ages. In contrast, beak size predicted less than 15% of beak shape variation in the 18.5-week old lateral lower mandibles and the lateral upper mandibles for both ages of turkeys. A recent study  showed sIn summary, landmark-based geometric morphometrics showed a range of phenotypic variation in the shape of dorsal upper mandibles, lateral lower mandibles, and lateral upper mandibles for domestic turkeys at 6 and 18.5 weeks of age. The main axes of shape variation were similar for the two ages, but the beak shape phenotypes of female and male turkeys differed significantly. The role of beak size in predicting beak shape for these turkeys differed between the three analyses and the two age groups. Given the wide phenotypic variation seen in turkey beak shape within this study, the beak shape variables could potentially be used to perform a quantitative genetic analysis to determine the heritability of beak shape variation.However, the implications from this turkey beak shape analysis is partially limited from only examining male-line turkeys and the reduced sample size for the shape analysis at 18.5 weeks of age. It is unclear if selection pressure for male-line traits, such as larger body weights and fast growth, could have influenced beak shape variation in comparison to female-line turkeys, which are selected for improved fertility, egg production, and egg hatchability . To fullSubsequent morphometric studies of turkey beak shape variation should analyze the lateral upper and lower mandible shape together to fully understand how the complete shape of the beak varies within domestic turkeys. Before moving forward with selective breeding for beak shape, future research should also examine if distinct beak shape phenotypes influence the feeding behaviour and efficiency of domestic turkeys. Furthermore, there is a need for research to determine the potential capacity for the different beak shape phenotypes to create skin and tissue damage when performing injurious pecking before genetic selection could be considered a realistic alternative to beak treatment in turkeys.S1 Dataset(XLSX)Click here for additional data file.S2 Dataset(XLSX)Click here for additional data file.S3 Dataset(XLSX)Click here for additional data file.S4 Dataset(XLSX)Click here for additional data file.S5 Dataset(XLSX)Click here for additional data file.S6 Dataset(XLSX)Click here for additional data file.


Granulocyte colony-stimulating factor (G-CSF)-mobilized peripheral blood stem cells (G-PBSC) has largely replaced unstimulated bone marrow (un-BM) for allografting because of accelerated engraftment, but with a higher morbidity and mortality of graft-versus-host-disease (GVHD). Recent studies suggested that G-CSF-primed BM (G-BM) had similar engraftment but lower morbidity and mortality of GVHD comparing to G-PBSC. A prospective, randomized, multicenter study was conducted to compare G-BM with G-PBSC as the grafts in allogeneic hematopoietic stem cell transplantation  for acute leukemia in first complete remission (CR1).Totally 101 adult leukemia in CR1 undergoing HLA-identical sibling transplants were randomized into G-BM or G-PBSC group. The primary study endpoint was GVHD-free/relapse-free survival (GRFS).P = 0.412, P = 0.39). G-BM group showed significantly lower II–IV acute GVHD (aGVHD) and similar III–IV aGVHD compared with G-PBSC group . The overall cumulative incidence of chronic GVHD (cGVHD) at 3 years were 22.3% ± 6.3% and 44.8% ± 7.6% (P = 0.026), respectively, and extensive cGHVD were 4.5% ± 3.1% and 15% ± 5.3% (P = 0.08), respectively, in G-BM and G-PBSC groups. Two groups had similar 3-year relapse, transplant-related mortality (TRM), overall survival (OS), and disease-free survival (DFS) . G-BM group showed significantly higher probability of GRFS than G-PBSC group . Graft content analysis revealed statistically higher frequency of myeloid-derived suppressor cells (MDSCs) in the G-BM than in G-PBSC grafts (P < 0.01), and recipients received statistically higher numbers of MDSCs in G-BM than in G-PBSC group (P = 0.045). Numbers of MDSCs infused to patients were negatively correlated with the severity of aGVHD . Multivariate analysis showed that MDSC cell dose below the median , recipient age , and high risk of disease  were independent risk factors for GRFS.Both the engraftment of neutrophil and platelet were 2 days later in G-BM than in G-PBSC group (G-BM grafts lead a better GRFS and less GVHD associated with a higher MDSCs content compared with G-PBSC grafts. Granulocyte colony-stimulating factor (G-CSF)-mobilized peripheral blood stem cells (G-PBSC) has replaced bone marrow (BM) as the most commonly used source of hematopoietic stem cells (HSCs) because of faster engraftment and practicability as well as acceptable graft-versus-host disease (GVHD) . Even th+CD25+ regulatory T (Treg) cells and tolerogenic dendritic cell differentiation [The mechanisms of G-CSF-primed grafts inducing immune tolerance are extensively studied, which include the induction of T helper type 2 (Th2) cell polarization and increase of CD4ntiation . But, th+ cells, different T cell subtypes and natural killer (NK) cells and, possibly, other cellular as well as cytokine components, which affect the time frame of engraftment and incidence of GVHD [Each graft source has its unique compositions, such as the number of CD34 of GVHD , 10. Mye of GVHD , 12. Rec of GVHD . In huma of GVHD . Our pre of GVHD .Based on these study results, we hypothesized that G-BM grafts lead a better GVHD-free/relapse-free survival (GRFS) and less GVHD associated with a higher MDSC content compared with G-PBSC grafts except for T cells and other compositions. And MDSCs might play the immunoregulatory role in GVHD. To identify this hypothesis, a randomized multicenter clinical trial was conducted.This prospective, randomized, open-label study was conducted from February 2013 to April 2015 in Nanfang Hospital, Southern Medical University, the First People’s Hospital of Chenzhou, People’s Hospital of Zhongshan City, and Concord Hospital of the Fujian Medical University. One hundred one patients with acute leukemia in first complete remission (CR1) undergoing allogeneic stem cell transplantation from an HLA-identical sibling were enrolled in this trial of G-BM or G-PBSC as the source of stem cells. The study was performed in accordance with modified Helsinki Declaration, and the protocol was approved by respective ethical review boards before study initiation. All patients and donors provided written informed consent.As we described previously , two myePneumocystis carinii. Acyclovir and ganciclovir were given for prophylaxis and treatment of cytomegalovirus (CMV) infection as prescribed in previous literature [9/L or absolute neutrophil count (ANC) reached 0.5 × 106/L. Patients received red blood cells and platelet transfusions if hemoglobin levels were ≤70 g/L and platelet count ≤20.0 × 109/L.As described previously , low molterature . Antifun8 total nucleated cells/kg or 2 × 106 CD34+ cells/kg recipient ideal body weight was collected. G-PBSC harvest was performed from day 5 of G-CSF to obtain 6.0 × 108 total nucleated cells/kg or a minimum CD34+ cell count of 3 × 106/kg recipient ideal body weight.All donors received G-CSF  5 μg/kg/day for 5 days. G-BM harvest was performed on the fifth day . At least 3.0 × 10+ cells, CD3+ T cells and subpopulation , CD19+B cells, CD3−CD56+ NK cells, and MDSCs by flow cytometry using standard procedures. MDSCs were defined as Linlow/negHLA-DR−CD33+CD11b+. All results were assayed by BD FACSCanto TMII (BD Biosciences), and the acquired data were further analyzed using BD-FACSDiva Software. Flow cytometric results were represented as percentage positive.Two grafts were analyzed in terms of CD34+ lymphocytes and MDSCs were isolated from G-BM or G-PBSC grafts by CD3 and HLA-DR, CD33 microbeads , according to manufacturer’s instructions. Purity of the selected populations was evaluated by flow cytometry, demonstrating an efficiency of separation above 95% in all experiments. Purified CD3+ T lymphocytes were labeled with 5 μM 5, 6-carboxy-fluorescein diacetate succinimidyl ester (CFSE) (Biolegend) and thereafter were co-cultured with isolated MDSCs at a MDSCs-to-CD3+ T cell ratio of 1:1 and 1:5 in the presence of anti-CD2/CD3/CD28 biotin beads (Miltenyi Biotec) in RPMI 1640 (Sigma-Aldrich) supplemented with 10% FBS, penicillin/streptomycin, and 2 mM l-glutamine. For positive control, lymphocytes were stimulated in the absence of MDSCs; for negative control, CD3+ cells and MDSCs were incubated without the stimuli. After 4 days of culture, the cells were harvested for CFSE dilution analysis by flow cytometry.Purified CD39/L and the first of 3 days with an absolute platelet count exceeding 20 × 109/L without transfusion support. Complete chimerism was defined as >95% donor cells detected. aGVHD was graded according to standard criteria [The primary study endpoint was GRFS. Secondary study endpoints were hematopoietic engraftment, acute and chronic GVHD, early infections, disease relapse, transplant-related mortality (TRM), overall survival (OS), and disease-free survival (DFS). GRFS was defined as absence of grade III–IV aGVHD, systemic immunosuppressive therapy requiring cGVHD, relapse, or death for any causes after allogeneic hematopoietic stem cell transplantation  , 18. Hemcriteria , and cGVU test. Numerical variables were analyzed as categories based on their values being below or above the median of the entire cohort. Correlation analysis between categorical variables was calculated using Spearman’s correlation test. Incidence of time-dependent outcomes was estimated by the method of Kaplan-Meier and compared by the log-rank test. Cumulative incidence curves in a competing risks setting were used to calculate probabilities of relapse and TRM. Cumulative incidence of relapse were calculated by regarding death as the competing event, whereas estimates of TRM were calculated by regarding disease relapse as the competing event [+ T cell dose, Treg cell dose, and MDSC dose. All statistical tests were two-sided, and a P value less than 0.05 was used to indicate statistical significance.Analysis was performed on October 31, 2016. Comparisons of categorical variables were made by means of chi-squared and Fisher’s exact tests for small numbers. Differences between numerical variables were calculated by means of the Mann-Whitney ng event . Gray teng event , 23. Coxn = 65) and acute lymphoblastic leukemia . Patients were all in CR1 and received grafts from matched siblings. The characteristics of patients, donors, and transplants are summarized in Table P > 0.05). G-PBSC recipients received statistically higher numbers of total nucleated cells (TNC), CD34+ cells, and about 4-fold more CD3+ T cells per kilogram than G-BM recipients (Table P < 0.01).A total of 101 patients were enrolled, with 49 randomized to G-BM and 52 to G-PBSC group. The primary diseases included acute myelogenous leukemia . After 5 days mobilization, the frequency of MDSCs in the BM increased to 0.65% ± 0.29%, compared with 0.41% ± 0.17% in the G-PB . Both in the BM and PB, the frequencies of MDSCs significantly increased after G-CSF treatment .To identify whether G-CSF induces the expansion of MDSCs in vivo, 20 donors were studied for the frequencies of MDSCs in the BM and PB before and after G-CSF mobilization (day 5). MDSCs were quantified with the following gating strategy Fig. . The res+ T cells for 4 days in the presence of T cell stimulators. The results showed that there was a significant inhibition of T cell proliferation in co-culture with MDSCs from the two grafts, which indicated that MDSCs from both G-BM and G-PBSC grafts were immunosuppressive .To verify whether G-CSF inducing MDSCs could be defined as functional MDSCs, we determined their immunosuppressive activity. Highly purified MDSCs from G-BM and G-PBSC grafts were, respectively, co-cultured with autologous CFSE-labeled CD3ive Fig. . Further+ T cells and subsets, CD19+ B cells, and CD3−CD56+ NK cells in the grafts for all the donors  were summarized in Table + T cells among lymphocytes was significantly lower in the G-BM than that in the G-PBSC grafts (P = 0.007). And the frequency of MDSCs was statistically higher in the G-BM than in the G-PBSC grafts . The percentage of Treg cells in the G-BM was also higher than in the G-PBSC though without statistical significance (P = 0.192). The percentage of other cell subsets in the two groups did not differ significantly . The absolute numbers of MDSCs infused in the G-BM group was 1.38–7.31 × 106/kg  compared with 1.65–5.98 × 106/kg  in the G-PBSC group (P = 0.045). Cell doses of other compositions including CD3+ T cells were all significantly lower in the G-BM than in the G-PBSC group, which were partly associated with significantly less TNC infusion in G-BM than in G-PBSC group.The percentage and absolute numbers of MDSCs, Treg cells and CD3P = 0.412), in the G-BM and G-PBSC groups. Median time to platelet reconstruction was 15 days  and 13 days , respectively (P = 0.390), in the G-BM and G-PBSC groups. To compare with unstimulated BM, the historical data of 32 patients who underwent unstimulated BM transplantation in our institutions between 1997 and 2003 was analyzed. The results showed that both the neutrophil and platelet reconstruction were significantly faster in the G-BM and G-PBSC than those in the unstimulated BM  with a median time of 22 days  to neutrophil reconstruction and 27 days  to platelet reconstruction in unstimulated BM group.Analyses of chimerism revealed that all patients obtained full donor chimerism by day +30 post-transplantation. Median time to neutrophil reconstruction was 14 days  and 12 days , respectively (P = 0.048), and III–IV aGVHD was 4.1% ± 2.8% in the G-BM group compared to 9.6% ± 4.1% in the G-PBSC group (P = 0.267). Ninety-five patients were alive at day +100 post-transplantation, cGVHD occurred in 10 patients in the G-BM group, and 20 patients in the G-PBSC group, including 4 cases and 8 cases persistent from aGVHD, respectively. The overall cumulative incidences of cGVHD at 3 years were 22.3% ± 6.3% and 44.8% ± 7.6% (P = 0.026), respectively, and extensive cGHVD were 4.5% ± 3.1% and 15% ± 5.3% (P = 0.08), respectively, in G-BM and G-PBSC groups. Two patients died from cGVHD in G-PBSC group, while no patients died from cGVHD in G-BM group (P > 0.05).The comparison with historical data of 32 patients who underwent unstimulated BM transplantation in our institutions was also conducted. The results showed a trend of lower incidence of GVHD in G-BM comparing to that in unstimulated BM group .Six patients developed II–IV aGVHD in the G-BM group, 15 patients in the G-PBSC group. The cumulative incidence of II–IV aGVHD was 12.2% ± 4.7% in the G-BM group compared to 28.8% ± 6.3% in the G-PBSC group . Similarly, patients who developed no extensive cGVHD received significantly higher numbers of MDSCs comparing with patients who developed . Correlation analysis showed a significantly negative correlation between the numbers of MDSCs infused and the severity of aGVHD . And a trend for negative correlation between MDSCs infusion and the severity of cGVHD was also found though without significant difference .The impact of graft contents on the incidence of GVHD was further analyzed. The results showed that patients who developed no II–IV aGVHD received statistically higher numbers of MDSCs than patients who did , similarly, no significant correlation was found between the numbers of other cellular components and acute or chronic GVHD .While cell doses of other compositions including CD3P > 0.05). One patient died from bacterial infection in G-BM group, while no patient died from infections in G-PBSC group (P = 1.000). Within 6 months post-transplantation, 14 (28.6%) patients in G-BM group and 16 (30.8%) patients in G-PB group had CMV viremia (P = 0.808); 3 patients in G-BM group and 5 patients in G-PB group had EBV viremia ( P = 1.000).Within the first 100 days post-transplantation, 55 patients developed 86 episodes of bacterial and/or fungal infections. There were 12 and 10 cases of bacterial infections, 7 and 5 cases of fungal infections, and 10 and 11 cases of bacterial and fungal mixed infections, respectively, in G-BM and G-PBSC groups , GVHD (n = 1 vs 3), and infections (n = 3 vs 3). Ten patients died of TRM, with 4 patients in G-BM group, and 6 patients in G-PBSC group (P = 0.333). Nineteen patients experienced relapse, as a result of 9 in G-BM group and 10 in G-PBSC group. Of the 19 relapse patients, 3 patients abandoned treatment and the other 16 patients were treated with chemotherapy, donor lymphocyte infusion, or second allo-HSCT. Seven cases achieved CR after treatment.With a median follow-up of 701 days post-transplantation, 76 patients were alive. Eleven patients in the G-BM and 14 in the G-PBSC group died. Causes of death included leukemia relapse (P = 0.840), and TRM were 8.2% ± 4.0% and 11.5% ± 4.5%, respectively, in the G-BM and G-PBSC groups . The 3-year cumulative OS were 75.5% ± 6.7% and 69.6% ± 7.1%, respectively, in the G-BM and G-PBSC groups , and DFS were 72.9% ± 6.4% and 67.1% ± 6.6%, respectively, in the G-BM and G-PBSC groups . The G-BM group showed significantly higher probability of GRFS than the G-PBSC group  was associated with a lower DFS. MDSC cell dose below the median of 3.65 × 106 cells/kg , older recipient age , and high risk of disease  were significantly associated with a lower GRFS. All factors studied were not significantly associated with relapse or OS in the univariate and multivariate analyses  , which indicated that higher content of MDSCs did not increase the incidence of relapse in this study.Risk factors for survival and relapse are presented in Table In the past two decades, unstimulated BM grafts have lost its use as the optimal source of HSCs because G-PBSC grafts has faster hematopoietic reconstruction. However, G-PBSC grafts have its shortage of increased GVHD compared with unstimulated BM grafts. Recently, some studies suggested that G-BM grafts had similar hematopoietic reconstruction, and less cGVHD, compared with G-PBSC grafts. Serody et al.  and MortA variety of factors affect the incidence of GVHD, such as graft source, donor source, HLA compatibility, and GVHD prophylaxis –31. Eachl-arginine depletion by arginase 1 and the inducible nitric oxidase (iNOS), generation of reactive oxygen species, release of transforming growth factor-beta (TGF-beta) and IL-10, cysteine sequestration, and regulatory T (Treg) cell induction [G-CSF induces immune tolerance through a variety of mechanisms including modulation on graft contents , 35. MDSnduction , 39–41. nduction  and Vendnduction  showing It is known that GVL effect mainly depends on T cell activity. So, we reasoned whether the higher MDSC content in the G-BM grafts increase the incidence of leukemia relapse because of its probable downregulated T cell responses. Animal models had demonstrated that the adoptive transfer of MDSCs can result in the successful control of GVHD without compromising GVT effects , 42. In GRFS is the primary endpoint of this research. Our results revealed that G-BM grafts showed significantly lower GVHD but similar hematopoietic reconstruction as well as relapse compared with G-PBSC grafts. So we questioned whether these results come to superior GRFS. As a result, the G-BM group showed significantly higher probability of 1-, 2-, and 3-year GRFS than the G-PBSC group. Considering the small sample size of this study, larger clinical trials need to be conducted as well as with further follow-up is wanted. In addition, BM grafts contain stromal cells such as mesenchymal stem cells (MSC), which show immunomodulatory effect on various immune cells. Whether stromal cells in G-BM grafts is associated with its lower GVHD compared with G-PBSC grafts need further research.G-BM grafts lead a better GRFS and less GVHD associated with a higher MDSC content compared with G-PBSC grafts. And MDSCs might play the immunoregulatory role in GVHD.