Network Pharmacology, Molecular Docking, and Experimental Validation on Guiluoshi Anzang Decoction Against Premature Ovarian Insufficiency.

BACKGROUND AND OBJECTIVES: Premature Ovarian Insufficiency (POI) is a disease suffered by women under the age of 40 when ovarian function has declined, seriously affecting both the physical and mental health of women. Guiluoshi Anzang decoction (GLSAZD) has been used for a long time and has a unique therapeutic effect on improving ovarian function. This study aims to investigate the mechanism of GLSAZD in treating POI through network pharmacology, molecular docking, and experimental verification. METHODS: In this study, the active ingredients of Guiluoshi Anzang Decoction and the targets of POI were obtained from TCMSP, BATMANN-TCM, Uniprot, GeneCards, and other databases, and network pharmacology analysis was performed. Molecular docking was conducted to validate the affinity of the main active ingredient of GLSAZD to key POI targets. A POI SD rat model was established, and HE staining, ELISA, Real-time PCR, and Western blot experiments were performed to verify the predicted core targets and the therapeutic effects. RESULTS: 10 core targets and the top 5 ingredients were screened out. Molecular docking showed core targets AKT1, CASP3, TNF, TP53, and IL6 had stable binding with the core 5 ingredients quercetin, kaempferol, beta-sitosterol, luteolin, and Stigmasterol. GO and KEGG enrichment analysis demonstrated the mechanism involved in the positive regulation of gene expression, PI3K-AKT signaling pathway, and apoptosis signaling pathways. Animal experiments indicated GLSAZD could up-regulate the protein expression of p-PI3K and p-AKT1 and the mRNA expression of STAT3 and VEGF, down-regulate TP53 and Cleaved Caspase-3 protein expression in rat`s ovarian tissues and serum TNF-α and IL-6 protein levels, activate PI3K-AKT signaling pathway and inhibit the apoptosis signaling pathway. CONCLUSION: GLSAZD treats POI through multi-component, multi-target, and multi-pathway approaches. This study provided evidence for its clinical application in treating POI and shed light on the study of traditional medicine of the Guangxi Zhuang Autonomous Region in China.

Cardiovascular mortality by cancer risk stratification in patients with localized prostate cancer: a SEER-based study.

Purpose: The risk of cardiovascular disease (CVD) mortality in patients with localized prostate cancer (PCa) by risk stratification remains unclear. The aim of this study was to determine the risk of CVD death in patients with localized PCa by risk stratification. Patients and methods: Population-based study of 340,806 cases in the Surveillance, Epidemiology, and End Results (SEER) database diagnosed with localized PCa between 2004 and 2016. The proportion of deaths identifies the primary cause of death, the competing risk model identifies the interaction between CVD and PCa, and the standardized mortality rate (SMR) quantifies the risk of CVD death in patients with PCa. Results: CVD-related death was the leading cause of death in patients with localized PCa, and cumulative CVD-related death also surpassed PCa almost as soon as PCa was diagnosed in the low- and intermediate-risk groups. However, in the high-risk group, CVD surpassed PCa approximately 90 months later. Patients with localized PCa have a higher risk of CVD-related death compared to the general population and the risk increases steadily with survival (SMR = 4.8, 95% CI 4.6-5.1 to SMR = 13.6, 95% CI 12.8-14.5). Conclusions: CVD-related death is a major competing risk in patients with localized PCa, and cumulative CVD mortality increases steadily with survival time and exceeds PCa in all three stratifications (low, intermediate, and high risk). Patients with localized PCa have a higher CVD-related death than the general population. Management of patients with localized PCa requires attention to both the primary cancer and CVD.

Evaluation on a teaching software for removable partial denture framework design.

BACKGROUND: Removable partial dentures (RPDs) are widely used as a dental prosthesis and have a wide application scope. OBJECTIVE: To explore the effect of using design software in the preclinical teaching of removable partial dentures (RPDs). METHODS: Unreal Engine software was used to build the RPD framework design teaching and training software. All 131 undergraduate students majoring in stomatology in the class of 2018, Kunming Medical University, were randomly divided into three groups and received either traditional experiment teaching, flipped classroom teaching, or software teaching for RPD design. The application effect of the software in the preclinical teaching of RPD design was evaluated by analyzing the examination results and through the use of a questionnaire survey. RESULTS: The differences in the theoretical examination scores among the traditional teaching group, the flipped classroom group, and the software teaching group were not statistically significant (P> 0.05), while the average design scores of upper Kennedy Class I and lower Kennedy Class II subclass II in the software teaching group were significantly higher than those in the traditional teaching group (P< 0.05). Overall, 75% of the students in the software teaching group reported that this teaching method could improve their learning initiative, a higher percentage than in the traditional teaching group (55.8%, P< 0.05). Meanwhile, 90.9% of the students in the software teaching group reported that the software could make RPD-related theoretical knowledge more visual and intuitive, and 93.2% of these students felt it was helpful for understanding the RPD three-dimensional (3D) spatial structure. These percentages were higher than those in the traditional teaching and flipped classroom groups (P< 0.05). CONCLUSION: In the preclinical teaching of RPD design, software training helped the students better understand the 3D structure of RPDs and establish clear design ideas, and it may also be valuable for in-depth research and promotion purposes.

Enhanced human hematopoietic stem and progenitor cell engraftment by blocking donor T cell-mediated TNFα signaling.

Allogeneic hematopoietic stem cell transplantation (HSCT) is a curative therapy, but the large number of HSCs required limits its widespread use. Host conditioning and donor cell composition are known to affect HSCT outcomes. However, the specific role that the posttransplantation signaling environment plays in donor HSC fate is poorly understood. To mimic clinical HSCT, we injected human umbilical cord blood (UCB) cells at different doses and compositions into immunodeficient NOD/SCID/IL-2Rgc-null (NSG) mice. Surprisingly, higher UCB cell doses inversely correlated with stem and progenitor cell engraftment. This observation was attributable to increased donor cell-derived inflammatory signals. Donor T cell-derived tumor necrosis factor-α (TNFα) was specifically found to directly impair the survival and division of transplanted HSCs and progenitor cells. Neutralizing donor T cell-derived TNFα in vivo increased short-term stem and progenitor cell engraftment, accelerated hematopoietic recovery, and altered donor immune cell compositions. This direct effect of TNFα on transplanted cells could be decoupled from the indirect effect of alleviating graft-versus-host disease (GVHD) by interleukin-6 (IL-6) blockade. Our study demonstrates that donor immune cell-derived inflammatory signals directly influence HSC fate, and provides new clinically relevant strategies to improve engraftment efficiency during HSCT.

Differential Expression Profile of Immunological Cytokines in Local Ovary in Patients with Polycystic Ovarian Syndrome: analysis by Flow Cytometry.

OBJECTIVE: Immune dysregulation may play an important role in the pathogenesis of polycystic ovary syndrome (PCOS). The purpose of this study was to investigate the Th1 and Th2-related cytokine profile in local ovary of women with PCOS. STUDY DESIGN: The T lymphocytes of follicular fluid (FF) were obtained at the time of oocyte retrieval before in-vitro fertilization (IVF) in woman with or without PCOS. After culturing with PMA, Ionomycin and Golgi stop agent, cells were detected for the intracellular cytokine production by flow cytometry. The profile of Th1 (IFN-γ, IL-2) and Th2 (IL-4, IL-10) cytokines of CD3(+) CD4(+)T lymphocyte subsets were analyzed through invert gating. These cytokines in FF were also evaluated by ELISA. RESULTS: Flow cytometry analysis showed that the production of Th1 (IFN-γ, IL-2) cytokines in FF lymphocytes in PCOS patients were significantly higher than those in controls; ELISA result also demonstrated that the concentration of Th1 cytokines (IFN-γ, IL-2) in FF in PCOS patients is significantly increased compared with those in controls. CONCLUSION: It is concluded that the immune dominance of Th1 may be the immunological feature of the ovary in PCOS patients. It might participate in the immune pathogenesis in the ovary of PCOS patients. These results suggest that chronic inflammation maybe one of the underlying mechanism for the pathogenesis of PCOS.

Associations between the codon 72 polymorphism of the TP53 gene and the risk of recurrent implantation failure.

AIM: Recurrent implantation failure (RIF) is the most common cause of unsuccessful pregnancy after assisted reproductive techniques. The tumor protein P53 (TP53) codon 72 polymorphism (G-C transversion) has been explored in susceptibility to RIF, but inconclusive results have been reported. The aim of this article is to estimate the associations between the TP53 codon 72 polymorphism and the risk of RIF. MATERIALS AND METHODS: A comprehensive search for relevant articles was conducted. The odds ratios (ORs) and 95% confidence intervals (CIs) for CC + GC versus GG, CC versus GC + GG, CC versus GG, GC versus GG genotypes, and C versus G allele, were estimated. Publication bias was explored. Statistical analyses were performed using RevMan 5.2 and Stata 11.0 software. RESULTS: A total of five case-control studies in five articles with 417 RIF cases and 325 controls were included. An overall random effect OR of 1.20 (95% CI, 0.66-2.19; P = 0.55) in the dominant model (CC + GC vs GG) was found. The results suggested that a lack of increased or decreased risks were found in individuals who carried the CC homozygote and heterozygote GC, in comparison with the homozygote GG. However, in subgroup analysis by ethnicity, a significantly increased risk was observed among Latin Americans in the dominant model (OR, 1.56; 95% CI, 1.04-2.33; P = 0.03). CONCLUSIONS: This meta-analysis shows that the TP53 codon 72 polymorphism is not associated with RIF risk in the overall population; however, associations were found in the Latin American population.

Humanized Chronic Graft-versus-Host Disease in NOD-SCID il2rγ-/- (NSG) Mice with G-CSF-Mobilized Peripheral Blood Mononuclear Cells following Cyclophosphamide and Total Body Irradiation.

Chronic graft-versus-host disease (cGvHD) is the major source of late phase morbidity and mortality after allogeneic hematopoietic stem cell transplantation. Humanized acute GvHD (aGvHD) in vivo models using NOD-SCID il2rγ-/- (NSG) mice are well described and are important tools for investigating pathogenicity of human cells in vivo. However, there have been only few reported humanized cGvHD mouse models. We evaluated if prolonged inflammation driven by low dose G-CSF-mobilized human PBMCs (G-hPBMCs) would lead to cGvHD following cyclophosphamide (CTX) administration and total body irradiation (TBI) in NSG mice. Engraftment was assessed in peripheral blood (PB) and in specific target organs by either flow cytometry or immunohistochemistry (IHC). Tissue samples were harvested 56 days post transplantation and were evaluated by a pathologist. Some mice were kept for up to 84 days to evaluate the degree of fibrosis. Mice that received CTX at 20mg/kg did not show aGvHD with stable expansion of human CD45+ CD3+ T-cells in PB (mean; 5.8 to 23.2%). The pathology and fibrosis scores in the lung and the liver were significantly increased with aggregation of T-cells and hCD68+ macrophages. There was a correlation between liver pathology score and the percentage of hCD68+ cells, suggesting the role of macrophage in fibrogenesis in NSG mice. In order to study long-term survival, 6/9 mice who survived more than 56 days showed increased fibrosis in the lung and liver at the endpoint, which suggests the infiltrating hCD68+ macrophages may be pathogenic. It was shown that the combination of CTX and TBI with a low number of G-hPBMCs (1x106) leads to chronic lung and liver inflammation driven by a high infiltration of human macrophage and mature human T cells from the graft, resulting in fibrosis of lung and liver in NSG mice. In conclusion this model may serve as an important pre-clinical model to further current understanding of the roles of human macrophages in cGvHD.

The codon 72 polymorphism of the TP53 gene and endometriosis risk: a meta-analysis.

Endometriosis is a chronic, inflammatory and common gynaecological disease. This study investigated the association between TP53 codon 72 polymorphism and the risk of endometriosis. A search for relevant articles was conducted in PubMed, Embase, CNKI, Wanfang, Weipu databases and Google Scholar. The strength of the relationships between TP53 codon 72 polymorphism and the risk of endometriosis was assessed by odds ratios (OR) and with 95% confidence intervals (CI). Sixteen case-control studies in 15 articles were included. Significant association was found in the dominant model (CC + GC versus GG) with an OR of 1.38 and 95% CI (1.14, 1.67). The results suggested that individuals who carried CC homozygote and heterozygote GC might have a 38% increased endometriosis risk when compared with the homozygote GG. In the subgroup analysis by ethnicity, significantly increased risk was observed among Asians (OR = 1.62, 95% CI = 1.18-2.23, P = 0.003) and Latin Americans (OR = 1.54, 95% CI = 1.16-2.03, P = 0.002) but not in Caucasians (OR = 1.02, 95% CI = 0.80-1.30) for the dominant model. The current meta-analysis suggested that TP53 codon 72 polymorphism was associated with the endometriosis risk, especially in Asians and Latin Americans.

Dynamic changes in endothelial cell adhesion molecule nepmucin/CD300LG expression under physiological and pathological conditions.

Vascular endothelial cells often change their phenotype to adapt to their local microenvironment. Here we report that the vascular endothelial adhesion molecule nepmucin/CD300LG, which is implicated in lymphocyte binding and transmigration, shows unique expression patterns in the microvascular endothelial cells of different tissues. Under physiological conditions, nepmucin/CD300LG was constitutively and selectively expressed at the luminal surface of the small arterioles, venules, and capillaries of most tissues, but it was only weakly expressed in the microvessels of the splenic red pulp and thymic medulla. Furthermore, it was barely detectable in immunologically privileged sites such as the brain, testis, and uterus. The nepmucin/CD300LG expression rapidly decreased in lymph nodes receiving acute inflammatory signals, and this loss was mediated at least in part by TNF-α. It was also down-regulated in tumors and tumor-draining lymph nodes, indicating that nepmucin/CD300LG expression is negatively regulated by locally produced signals under these circumstances. In contrast, nepmucin/CD300LG was induced in the high endothelial venule-like blood vessels of chronically inflamed pancreatic islets in an animal model of non-obese diabetes. Interestingly, the activated CD4(+) T cells infiltrating the inflamed pancreas expressed high levels of the nepmucin/CD300LG ligand(s), supporting the idea that nepmucin/CD300LG and its ligand interactions are locally involved in pathological T cell trafficking. Taken together, these observations indicate that the nepmucin/CD300LG expression in microvascular endothelial cells is influenced by factor(s) that are locally produced in tissues, and that its expression is closely correlated with the level of leukocyte infiltration in certain tissues.

A novel monoclonal antibody against cannabinoid receptor 1.

The cannabinoid receptor 1 (CBR1) is being widely investigated because of its specific structure and functions compared with other cannabinoid receptors. In this study, we immunized BALB/c mice with synthesized human CBR1 polypeptide and obtained a novel monoclonal antibody (MAb) against human CBR1. Analysis through enzyme-linked immunosorbent assay (ELISA), spot-ELISA, Western blot, and immunohistochemistry revealed that the MAb was specifically against recombinant human CBR1 protein, and its subtype and affinity constant (Kaff) were IgG2b/k and 7.85 × 10(8) M/L, respectively. Using this MAb we found that CBR1 is expressed on HL-7702 cells and lipid tissue, raising the possibility that the CBR1 may take a role in glucose and lipid metabolism. Thus, this antibody might facilitate studies for pathophysiology of diseases associated with glucose and lipid metabolism abnormality.

Predictive factors for acute renal failure in crush injuries in the Sichuan earthquake.

INTRODUCTION: The Sichuan earthquake caused a large number of crush injuries and many of them developed acute renal failure (ARF). A retrospective study was performed on victims with crush injuries of West China Hospital to investigate the predictive factors for acute renal failure (ARF) in crush injuries. PATIENTS AND METHODS: Medical records of injured victims treated in West China Hospital within the first week after the Sichuan earthquake were retrospectively reviewed and 101 patients with crush injury were enrolled in the study. We divided them into an ARF group and a non-ARF group. The clinical data of included patients were extracted and analysed. RESULTS: Patients with ARF accounted for 42% of the included population. Patients younger than 20 made up the biggest age category (45%), and the entrapped time under the debris (22 [IQR 3.5-38]h) was longer than previous reports. In univariate analysis, male gender, multiple crush injuries, medical comorbidities, surgical interventions and infections were more frequent in patients with ARF than in those without ARF. Mean arterial pressure was higher in the ARF group. Besides, the risk of ARF was increased by creatine kinase >14,494.5IU/L most significantly, followed by time under the rubble >4h, aspartate transaminase >453.5IU/L, albumin <27.15g/L and white blood cell >11.8×10(9)/L. In multivariate analysis, male gender, time under the rubble, multiple crush injuries, surgical interventions, infections and creatine kinase level were independently associated with ARF in crush injuries. CONCLUSIONS: The entrapped time under the debris, multiple crush injuries, male gender, infections, and creatine kinase level are predictive factors for ARF in crush injuries.

[Comparision of intracellular localization and recycling route of mouse nepmucin and CD31 in endothelial cells].

AIM: To compare the localization and recycling between nepmucin and CD31 molecules on transfected endothelial cells, and attempted to clarify the recycling mechanisms of nepmucin in endothelial cells. METHODS: Recycling assay and internalization assay were employed to compare the localization and recycling pathway of nepmucin and CD31. The internalized and recycling nepmucin and CD31 molecules on transfected endothelial cells were double or single stained with specific fluorchrome-labeled monoclonal antibodies against nepmucin (Alexa Fluor 488-ZAQ5) and/or CD31 (Alexa Fluor 488-anti-CD31 or Alexa Fluor 594-anti-CD31), then observed under confocal microscopy. RESULTS: Mouse nepmucin underwent intracellular recycling like CD31, but which recycling rate was significantly lower. The CD31 and nepmucin molecules showed largely distinct localization in endothelial cells. CD31 was found mainly on the cell surface, while nepmucin was found predominantly in the deep area of cytoplasm and partly on the cell membrane. CONCLUSION: The distribution of mouse nepmucin in endothelial cells are different from CD31. Nepmucin underwent intracellular recycling like CD31 but employed different mechanisms.

Overexpression of the c-Myb but not its leukemogenic mutant DNA-binding domain increased adipogenic differentiation in mesenchymal stem cells.

The c-Myb protein is a vital transcription factor that regulates the differentiation of hematopoietic cells. Previous works have noticed that c-Myb is involved in an epigenetic control mechanism, in which the c-Myb DNA-binding domain (DBD) binds to the N-terminal histone tail of H3 to facilitate it acetylation and activate endogenous differentiation genes, while the leukemogenic mutant of c-Myb does not have these functions. However, whether c-Myb has corresponding biologic functions on the differentiation of other cells except for hematopoietic cells has not been explored. In our studies, we constructed the c-Myb wild type and its leukemogenic mutant DBD recombinant adenovirus with replication-defective adenoviral vectors carrying the GFP gene. We compared their roles on adipogenic differentiation efficiency in human bone marrow-derived mesenchymal stem cells (hMSCs). Our results demonstrated that the overexpression of c-Myb could enhance adipogenic differentiation in hMSCs, while the overexpression of its leukemogenic mutant blocked the adipogenic differentiation to a certain extent. These suggest that c-Myb play an important role in the hMSCs differentiation too, which is consistent with the epigenetic control mechanism of c-Myb.

Effect of high glucose levels on the calcification of vascular smooth muscle cells by inducing osteoblastic differentiation and intracellular calcium deposition via BMP-2/Cbfα-1 pathway.

In this paper, we investigate the effect and the possible mechanism of high glucose levels on the calcification of human aortic smooth muscle cells (HASMCs). HASMCs were divided into four groups: normal glucose group (NG), osmolality control group (OC), high glucose group (HG, HASMCs culture medium containing 30 mmol/L glucose), and high glucose plus recombinant human Noggin protein (bone morphogenetic protein-2 (BMP-2) antagonist) group (HN). The mRNA levels and the protein expressions of BMP-2 and core binding factor alpha-1 (Cbfα-1) were measured by real-time quantitative polymerase chain reaction (PCR) and Western blot. After induced by 10 mmol/L ß-glycerol phosphoric acid, cells were harvested for assessments of alkaline phosphatase (ALP) activities at Days 1, 2, and 3, and intracellular calcium contents at Days 7 and 14, respectively. High glucose levels increased the mRNA levels and the protein expressions of BMP-2 and Cbfα-1 (P<0.05). The expression of Cbfα-1 was partially blocked by Noggin protein (P<0.05), while BMP-2 was not (P>0.05). After being induced by ß-glycerol phosphoric acid, high glucose levels increased the ALP activity [(48.63±1.03) vs. (41.42±2.28) U/mg protein, Day 3; P<0.05] and the intracellular calcium content [(2.76±0.09) vs. (1.75±0.07) µmol/mg protein, Day 14; P<0.05] in a time-dependent manner when compared with the NG group, while the ALP activity could not be blocked by Noggin protein [(48.63±1.03) vs. (47.37±0.97) U/mg protein, Day 3; P>0.05]. These results show that high glucose levels can evoke the calcification of HASMCs by inducing osteoblastic trans-differentiation and intracellular calcium deposition via the BMP-2/Cbfα-1 pathway, which can be partially blocked by Noggin protein.

Synthetic peptide coupled to KLH elicits antibodies against beta8 integrin.

The beta8 integrin, which forms alphavbeta8 heterodimers, is being widely investigated because of its specific structure and functions compared with other integrins. In this report, a 12 aa-long peptide of beta8 integrin cytoplasmic domain was synthesized according to a published sequence and covalently coupled to keyhole limpet hemocyanin (KLH). Three stable strains of hybridomas (3G6, 5C7, 5H3) that can secrete high specific monoclonal antibodies against beta8 integrin were successfully established by hybridoma technique. The isotypes of these MAbs were tested to be IgG2a. Their characterizations were shown by enzyme-linked immunosorbent assay (ELISA), Western blot analysis, and immunocytochemistry (ICC). The affinity constants (Kaff ) of the MAbs 3G6, 5C7, and 5H3 were measured by non-competitive ELISA respectively. Western blot analyses and immunocytochemistry demonstrated that all the MAbs were directed against beta8 integrin with high specificity.

Dermatopontin is expressed in human liver and is downregulated in hepatocellular carcinoma.

Dermatopontin (DPT) was recently found as a downstream target of vitamin D receptor, which is a key molecule in the 1,25-dihydroxy-vitamin D(3) anti-hepatoma proliferation pathway. MCTx-1 from Millepora, a homolog of DPT, is identified as a cytotoxin towards leukemia cells. The aim of this study was to analyze DPT expression in hepatocellular carcinoma (HCC) based on the analysis for DPT gene in normal tissues in order to estimate its function in the progression of HCC. DPT mRNA expression was analyzed in normal tissues and HCC cell lines by RT-PCR, and in HCC tissue by RT-PCR and real-time PCR. Its protein was examined in HCC tissues by Western blot and immunohistochemistry assays. Meanwhile, transforming growth factor-beta1 (TGF-beta1) that is closely associated with HCC and DPT was observed by immunohistochemistry in HCC tissues. The results showed that DPT mRNA was strongly expressed in human fetal and adult liver, kidney, and spleen, weakly in ovary and heart, and absent in other tissues and HCC cell lines examined. Its mRNA was significantly downregulated in HCC tissues, while its protein was weakly expressed in tumor compared with non-tumor. DPT is located mainly in the cytoplasm of several cell types in the liver; it has been identified also in the extracellular matrix of the skin. TGF-beta1 was observed in extensive tumor tissue of HCC. This fact suggests that DPT can play various roles in different tissues and might be a molecule related to carcinogenesis and the progression of HCC via possible interaction with TGF-beta1 and other potential mechanisms.

Characteristics of a novel monoclonal antibody against interleukin-14alpha.

Interleukin-14alpha is cytokine encoded by the plus strand of il-14 gene using exons 3-10. It has been demonstrated in an IL-14-transgenic mice model that IL-14alpha could enhance an immune response to both TD and TI antigens and induce a phenotype that is very similar to SLE and Sjögren's syndrome, indicating that IL-14alpha may play an important role in autoimmune disease. To further study the role of IL-14alpha in human immune diseases, there is an urgent need to generate specific anti-human IL-14alpha monoclonal antibodies. In this study, we immunized BALB/c mice with synthesized human IL-14alpha-C polypeptide and obtained a novel monoclonal antibody, the anti-IL-14alpha-C antibody. Analysis by enzyme-linked immunosorbent assay (ELISA), spot-ELISA, Western blot, and immuoprecipitation revealed that the MAb were specifically against recombinant human IL-14alpha protein, and its subtype and affinity constant (K(aff)) were IgG2a/kappa and 1.007 x 10(8) M(-1), respectively. This antibody might facilitate studies for pathophysiology of autoimmune disease.

Differential effects of granulocyte colony-stimulating factor on marrow- and blood-derived hematopoietic and immune cell populations in healthy human donors.

A recent phase III trial comparing granulocyte colony-stimulating factor (G-CSF)-stimulated bone marrow (G-BM) and G-CSF-mobilized peripheral blood (G-PB) in matched sibling allograft recipients showed that G-BM produced a similar hematologic recovery but a reduced incidence of extensive chronic graft-versus-host disease, indicating differences in the cell populations infused. As a first step toward identifying these differences, we treated a group of healthy adult humans with 4 daily doses of G-CSF 10 microg/kg and monitored the effects on various hematopoietic and immune cell types in the PB and BM over 12 days. G-CSF treatment caused rapid and large but transient increases in the number of circulating CD34+ cells, colony-forming cells, and long-term culture-initiating cells and in the short-term repopulating activity detectable in nonobese diabetic/severe combined immunodeficiency/beta2-microglobulin-null mice. Similar but generally less marked changes occurred in the same cell populations in the BM. G-CSF also caused transient perturbations in some immune cell types in both PB and BM: these included a greater increase in the frequency of naive B cells and CD123+ dendritic cells in the BM. The rapidity of the effects of G-CSF on the early progenitor activity of the BM provides a rationale for the apparent equivalence in rates of hematologic recovery obtained with G-BM and G-PB allotransplants. Accompanying effects on immune cell populations are consistent with a greater ability of G-BM to promote tolerance in allogeneic recipients, and this could contribute to a lower rate of chronic graft-versus-host disease.