1.Effect of removing microglia from spinal cord on nerve repair after spinal cord injury in mice.
Qi JIANG ; Chao QI ; Yuerong SUN ; Shiyuan XUE ; Xinyi WEI ; Haitao FU
Chinese Journal of Reparative and Reconstructive Surgery 2025;39(6):754-761
OBJECTIVE:
To investigate the effects of removing microglia from spinal cord on nerve repair and functional recovery after spinal cord injury (SCI) in mice.
METHODS:
Thirty-nine 6-week-old female C57BL/6 mice were randomly divided into control group ( n=12), SCI group ( n=12), and PLX3397+SCI group ( n=15). The PLX3397+SCI group received continuous feeding of PLX3397, a colony-stimulating factor 1 receptor inhibitor, while the other two groups were fed a standard diet. After 14 days, both the SCI group and the PLX3397+SCI group were tested for ionized calcium binding adapter molecule 1 (Iba1) to confirm that the PLX3397+SCI group had completely depleted the spinal cord microglia. The SCI model was then prepared by clamping the spinal cord in both the SCI group and the PLX3397+SCI group, while the control group underwent laminectomy. Preoperatively and at 1, 3, 7, 14, 21, and 28 days postoperatively, the Basso Mouse Scale (BMS) was used to assess the hind limb function of mice in each group. At 28 days, a footprint test was conducted to observe the gait of the mice. After SCI, spinal cord tissue from the injury site was taken, and Iba1 immunofluorescence staining was performed at 7 days to observe the aggregation and proliferation of microglia in the spinal cord. HE staining was used to observe the formation of glial scars at the injury site at 28 days; glial fibrillary acidic protein (GFAP) immunofluorescence staining was applied to astrocytes to assess the extent of the injured area; neuronal nuclei antigen (NeuN) immunofluorescence staining was used to evaluate neuronal survival. And 5-hydroxytryptamine (5-HT) immunofluorescence staining was performed to assess axonal survival at 60 days.
RESULTS:
All mice survived until the end of the experiment. Immunofluorescence staining revealed that the microglia in the spinal cord of the PLX3397+SCI group decreased by more than 95% compared to the control group after 14 days of continuous feeding with PLX3397 ( P<0.05). Compared to the control group, the BMS scores in the PLX3397+SCI group and the SCI group significantly decreased at different time points after SCI ( P<0.05). Moreover, the PLX3397+SCI group showed a further decrease in BMS scores compared to the SCI group, and exhibited a dragging gait. The differences between the two groups were significant at 14, 21, and 28 days ( P<0.05). HE staining at 28 days revealed that the SCI group had formed a well-defined and dense gliotic scar, while the PLX3397+SCI group also developed a gliotic scar, but with a more blurred and loose boundary. Immunofluorescence staining revealed that the number of microglia near the injury center at 7 days increased in the SCI group than in the control group, but the difference between groups was not significant ( P>0.05). In contrast, the PLX3397+SCI group showed a significant reduction in microglia compared to both the control and SCI groups ( P<0.05). At 28 days after SCI, the area of spinal cord injury in the PLX3397+SCI group was significantly larger than that in SCI group ( P<0.05); the surviving neurons significantly reduced compared with the control group and SCI group ( P<0.05). The axonal necrosis and retraction at 60 days after SCI were more obvious.
CONCLUSION
The removal of microglia in the spinal cord aggravate the tissue damage after SCI and affecte the recovery of motor function in mice, suggesting that microglia played a neuroprotective role in SCI.
Animals
;
Spinal Cord Injuries/surgery*
;
Microglia/pathology*
;
Female
;
Mice
;
Mice, Inbred C57BL
;
Nerve Regeneration/drug effects*
;
Spinal Cord/pathology*
;
Pyrroles/administration & dosage*
;
Aminopyridines/administration & dosage*
;
Recovery of Function
;
Disease Models, Animal
;
Calcium-Binding Proteins/metabolism*
;
Receptors, Granulocyte-Macrophage Colony-Stimulating Factor/antagonists & inhibitors*
;
Microfilament Proteins/metabolism*
;
Glial Fibrillary Acidic Protein/metabolism*
2.Effect of different culture time on immunomembrane proteins of human monocyte-derived dendritic cells and their exosomes.
Shumin LUO ; Fang XU ; Pengpeng LU ; Yiyue WANG ; Chuanyun LI ; Weihua LI
Chinese Journal of Cellular and Molecular Immunology 2025;41(11):971-977
Objective To investigate how culture duration affects the expression of immune membrane proteins in human monocyte-derived dendritic cells (DCs) and their exosomes (DEXs). Methods Human monocytes were induced with recombinant granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin 4 (IL-4) to differentiate into DCs and were subsequently matured with tumor necrosis factor α(TNF-α). Exosomes were isolated by ultracentrifugation, and DEXs were identified by transmission electron microscopy and Amnis imaging flow cytometry, which were also used to quantify the expression of immune membrane proteins on DCs and DEXs. Results On the 10th day of culture, DCs displayed high surface expression of CD11c, CD80, CD86, major histocompatibility complex class I (MHC-I), and MHC-II. Expression peaked at day 18(CD11c: 78.66%±20.33%, CD80: 76.41%±10.02%, CD86: 96.43%±0.43%, MHC-I: 84.71%±2.96%, MHC-II: 80.01%±7.03%). After day 24, the overall expression showed a declining trend, with statistically significant differences observed for all markers except CD80 and MHC-II. By day 30, 80% of the DCs still expressed CD80, CD86, and MHC-II. The expression of immune membrane proteins on DEX surfaces also reached its peak on day 18, followed by an overall decline with prolonged culture time, with statistically significant differences observed for all markers except CD80. Correlation analysis revealed a significant positive relationship between the expression levels of immune membrane proteins on DC and DEX surfaces (CD11c: r=0.98; CD80: r=0.65; CD86: r=0.82; MHC-I: r=0.86; MHC-II: r=0.93). Conclusion Human monocyte-derived DCs in vitro express high expression of immune membrane proteins and maintain stable expression over a specific period. The exosomes secreted by these cells similarly demonstrate high surface expression of immune membrane proteins, with temporal trends aligned with those of the parent DCs.
Humans
;
Dendritic Cells/immunology*
;
Exosomes/immunology*
;
Monocytes/metabolism*
;
Cells, Cultured
;
Time Factors
;
B7-1 Antigen/metabolism*
;
Membrane Proteins/immunology*
;
Cell Culture Techniques/methods*
;
B7-2 Antigen/metabolism*
;
Cell Differentiation
;
CD11c Antigen/metabolism*
;
Granulocyte-Macrophage Colony-Stimulating Factor/pharmacology*
3.Efficacy and Safety of DCAG Regimen in Patients with Relapsed/Refractory Acute Myeloid Leukemia.
Hui-Sheng ZHOU ; Yu-Qing LI ; Yu-Xin WANG ; Ya-Lei HU ; Kai-Li MIN ; Chun-Ji GAO ; Dai-Hong LIU ; Xiao-Ning GAO
Journal of Experimental Hematology 2025;33(1):9-19
OBJECTIVE:
To evaluate the efficacy and safety of DCAG (decitabine, cytarabine, anthracyclines, and granulocyte colony-stimulating factor) regimen in the treatment of patients with relapsed/refractory (R/R) acute myeloid leukemia (AML).
METHODS:
The clinical data of 64 R/R AML patients received treatment at Chinese PLA General Hospital from January 1st, 2012 to December 31st, 2022 were retrospectively analyzed. Primary endpoints included efficacy measured by overall response rate (ORR) and safety. Secondary endpoints included overall survival (OS), event-free survival (EFS) and duration of response (DOR). The patients were followed from enrollment until death, or the end of last follow-up (June 1st, 2023), whichever occurred first.
RESULTS:
Sixty-four patients who failed prior therapy were enrolled and completed 1 cycle, and 26 and 5 patients completed 2 and 3 cycles, respectively. Objective response rate was 67.2% [39: complete remission (CR)/CR with incomplete hematologic recovery (CRi), 4: partial remission (PR)]. With a median follow-up of 62.0 months (1.0-120.9), the median overall survival (OS) was 23.3 and event-free survival was 10.6 months. The median OS was 51.7 months (3.4-100.0) in responders (CR/CRi/PR) while it was 8.4 months (6.1-10.7) in nonresponders ( P <0.001). Grade 3-4 hematologic toxicities were observed in all patients. Four patients died from rapid disease progression within 8 weeks after chemotherapy.
CONCLUSION
The DCAG regimen represents a feasible and effective treatment for R/R AML.
Humans
;
Leukemia, Myeloid, Acute/drug therapy*
;
Cytarabine/administration & dosage*
;
Granulocyte Colony-Stimulating Factor/administration & dosage*
;
Retrospective Studies
;
Male
;
Female
;
Decitabine
;
Antineoplastic Combined Chemotherapy Protocols/therapeutic use*
;
Anthracyclines/administration & dosage*
;
Middle Aged
;
Adult
;
Treatment Outcome
;
Aged
;
Recurrence
4.The Influence of COVID-19 Infection on the Mobilization and Collection of Autologous Peripheral Blood Stem Cells in Patients with Multiple Myeloma.
Guo-Rong WANG ; Guang-Zhong YANG ; Yun LENG ; Yin WU ; Ai-Jun LIU ; Wen-Ming CHEN
Journal of Experimental Hematology 2025;33(2):455-462
OBJECTIVE:
To analyze the effect of COVID-19 infection on the mobilization and collection of autologous peripheral blood stem cells in patients with multiple myeloma.
METHODS:
The general baseline data, treatment factors before mobilization collection, collection status, and treatment overview after collection of autologous peripheral blood stem cells at Beijing Chaoyang Hospital affiliated with Capital Medical University from January 1, 2020 to July 15, 2023 were analyzed.
RESULTS:
269 patients underwent mobilization and collection of autologous peripheral blood stem cells. Among them, 32 cases with COVID-19 infection history (COVID-19 group) and 237 cases without COVID-19 infection history (non-COVID-19 group). In the COVID-19 group, 17 cases were treated with chemotherapy (etoposide)+G-CSF, and 15 cases were treated with plerixafor +G-CSF. In the non-COVID-19 group, 214 cases were treated with chemotherapy +G-CSF, 17 cases were treated with plerixafor +G-CSF, and 6 cases were treated with chemotherapy + plerixafor +G-CSF. The number of CD34+ cells, collection success rate, and excellence rate in the COVID-19 group and the non-COVID-19 group were [5.52 (0.94-26.87) vs 4.80 (0.53-37.20)]×106/kg (P =0.610), (93.8% vs 85.2%) (P =0.275), (62.5% vs 49.4%) (P =0.190), respectively. Among 113 patients mobilized with etoposide +G-CSF, the number of CD34+ cells, success rate, and excellence rate collected from COVID-19 infection (17 cases) and non-COVID-19 infection (96 cases) were [7.54 (2.66-26.87) vs 7.78 (2.26-37.20)]×106/kg (P =0.847), (100.0% vs 100.0%) (no P value), (82.4% vs 86.5%) (P =0.655), respectively. Among 32 patients mobilized by plerixafor +G-CSF, the number of CD34+ cells, success rate and excellence rate of COVID-19 infection (15 cases) and non-COVID-19 infection (17 cases) were [3.82 (0.94-7.27) vs 4.11 (0.53-9.05)]×106/kg (P =0.821), (86.7% vs 88.2%) (P =0.893), (40.0% vs 35.3%) (P =0.784), respectively. In 32 patients with COVID-19 infection, the number of CD34+ cells collected by etoposide +G-CSF (17 cases) and plerixafor +G-CSF (15 cases), as well as the success rate and excellence rate were [7.54 (2.66-26.87) vs 3.82(0.94-7.27)]×106/kg (P =0.004), (100.0% vs 86.7%) (P =0.120), (82.4% vs 40.0%) (P =0.014), respectively. By 2023.7.31, 232 patients (86.2%, 232/269) had received transplantation, including 24 patients in the COVID-19 group and 208 patients in the non-COVID-19 group. The median number of CD34+ cells infused in the two groups was [3.67 (2.50-13.44) vs 3.11(1.12-19.89)]×106/kg (P =0.058), the median days of neutrophil engraftment [11(9-13) vs 11(9-17)] (P =0.674), the median days of platelet engraftment [11(0-23), 12(0-43)] (P =0.279), respectively.
CONCLUSION
The history of COVID-19 infection did not affect the PBSC mobilization, collection and transplantation of patients with myeloma. In patients with COVID-19 infection, the results of chemotherapy mobilization with etoposide seems to be better than that of plerixafor mobilization, but further research is needed to clarify.
Humans
;
COVID-19/complications*
;
Multiple Myeloma/complications*
;
Hematopoietic Stem Cell Mobilization
;
Transplantation, Autologous
;
Granulocyte Colony-Stimulating Factor/therapeutic use*
;
Peripheral Blood Stem Cell Transplantation
;
SARS-CoV-2
;
Middle Aged
;
Peripheral Blood Stem Cells
;
Male
;
Female
;
Cyclams
;
Benzylamines
5.Clinical Efficacy of CAG Regimen Combined with Venetoclax, Chidamide, and Azacitidine in the Treatment of Elderly Patients with Acute Myeloid Leukemia.
Qing-Yang LIU ; Yu JING ; Meng LI ; Sai HUANG ; Yu-Chen LIU ; Ya-Nan WEN ; Jing-Jing YANG ; Wen-Jing GAO ; Ning LE ; Yi-Fan JIAO ; Xia-Wei ZHANG ; Li-Ping DOU
Journal of Experimental Hematology 2025;33(4):945-950
OBJECTIVE:
To explore the efficacy and adverse reactions of CAG regimen combined with venetoclax, chidamide, and azacitidine in the treatment of elderly patients with acute myeloid leukemia (AML).
METHODS:
15 elderly AML patients aged≥60 years old who were admitted to the Hematology Department of our hospital from May 2022 to October 2023 were treated with the CAG regimen combined with venetoclax, chidamide and azacitidine, and the efficacy, treatment-related adverse events, overall survival (OS) and event-free survival (EFS) were analyzed.
RESULTS:
After one course of treatment, 11 out of 15 patients achieved complete response (CR), 3 patients achieved CR with incomplete hematologic recovery (CRi), and 1 patient died due to prior infection before efficacy evaluation, and the overall response rate (ORR) was 93.3% (14/15). The median follow-up time was 131 (19-275) days, with median OS and EFS both remaining unreached. Next-generation sequencing (NGS) analysis showed that among the 15 patients, 13 were detected with gene mutations, and there were 7 genes with mutation frequencies of more than 10%, including ASXL1 (4 cases), RUNX1 (4 cases), BCOR (3 cases), DNMT3A (3 cases), STAG2 (2 cases), IDH1/2 (2 cases), and TET (2 cases). Among the 13 patients with detectable mutations, 12 patients achieved composite response (CR+CRi). The average recovery time of white blood cell count was 14.6 days after chemotherapy, and the average recovery time of platelets was 7.7 days after chemotherapy. The main adverse event was myelosuppression, with 10 patients accompanied by infection. Except for 1 patient who died due to septic shock during chemotherapy, no patients experienced serious complications such as heart, liver, or kidney damage during the treatment process.
CONCLUSION
The CACAG+V regimen, which combines the CAG regimen with venetoclax, chidamide, and azacitidine, can be applied in the treatment of elderly AML patients, demonstrating good safety and induction remission rate.
Humans
;
Leukemia, Myeloid, Acute/drug therapy*
;
Bridged Bicyclo Compounds, Heterocyclic/therapeutic use*
;
Sulfonamides/therapeutic use*
;
Aminopyridines/therapeutic use*
;
Antineoplastic Combined Chemotherapy Protocols/therapeutic use*
;
Azacitidine/therapeutic use*
;
Aged
;
Benzamides/therapeutic use*
;
Male
;
Female
;
Treatment Outcome
;
Middle Aged
;
Cytarabine
;
Aclarubicin
;
Granulocyte Colony-Stimulating Factor
6.Effective Salvage Mobilization of Peripheral Blood Stem Cells with High-Dose Etoposide in Newly Diagnosed Multiple Myeloma Patients Who Failed Initial Mobilization with High-Dose Cyclophosphamide.
Yue-Qi WANG ; Shi-Hua ZHAO ; Yi MA ; Xi-Lin CHEN ; Shun-Zong YUAN ; Na-Na CHENG ; Guang-Ning SHI ; Wen-Rong HUANG ; Xiu-Bin XIAO
Journal of Experimental Hematology 2025;33(5):1380-1385
OBJECTIVE:
To explore the safety and efficacy of high-dose etoposide (VP-16) combined with recombinant human granulocyte colony-stimulating factor (rhG-CSF) as salvage mobilization for peripheral blood stem cells (PBSC) in newly diagnosed multiple myeloma (NDMM) patients.
METHODS:
From April 2021 to May 2023, eight NDMM patients who had failed to yield sufficient PBSC during initial mobilization with high-dose cyclophosphamide (CTX) combined with rhG-CSF underwent salvage mobilization with 1.2 g/m2 etoposide combined with rhG-CSF 10 μg/(kg·d). The effects and adverse reactions of initial mobilization and salvage mobilization were analyzed.
RESULTS:
For salvage mobilization and initial mobilization, the numbers of PBSC collections were 16 and 18, respectively. The mean value of total collected CD34+ cells were (11.90±5.75)×106/kg and (1.67±0.75)×106/kg (P =0.0010) in salvage mobilization group and initial mobilization group, respectively. The proportion of patients with a total collection of CD34+ cell count≥2×106/kg were 100% and 37.5% (P =0.0625), and the proportion of patients with a total collection of CD34+ cell count≥5×106/kg were 87.5% and 0% (P =0.0156) in salvage mobilization group and initial mobilization group, respectively. For five patients who underwent high-dose CTX initial mobilization but had a total CD34+ cell count < 2×106/kg, successful collection was achieved through salvage mobilization with high-dose VP-16. Salvage mobilization with high-dose VP-16 was scheduled 2-3 weeks after failure of CTX mobilization. Adverse reactions of high-dose VP-16 mobilization did not increase compared to the initial mobilization with high-dose CTX.
CONCLUSION
As a salvage mobilization regimen, VP-16 1.2 g/m2 combined with rhG-CSF is safe and highly effective in NDMM patients who failed to initial mobilization with high-dose CTX combined with rhG-CSF.
Humans
;
Multiple Myeloma/therapy*
;
Etoposide/therapeutic use*
;
Hematopoietic Stem Cell Mobilization/methods*
;
Cyclophosphamide/therapeutic use*
;
Granulocyte Colony-Stimulating Factor
;
Salvage Therapy
;
Peripheral Blood Stem Cells
;
Male
;
Middle Aged
;
Female
;
Peripheral Blood Stem Cell Transplantation
7.Regulatory Effects of Cytokines on Spontaneous Pyroptosis in Neutrophils.
Tong CHEN ; Qian REN ; Feng-Xia MA
Acta Academiae Medicinae Sinicae 2025;47(4):497-508
Objective To explore the regulatory effects of cytokines interleukin(IL)-1β,IL-6,tumor necrosis factor alpha(TNF-ɑ),gamma interferon(IFN-γ),granulocyte colony-stimulating factor(G-CSF),and granulocyte-macrophage colony-stimulating factor(GM-CSF)on spontaneous pyroptosis in neutrophils.Methods Neutrophils isolated from mouse bone marrow by density-gradient centrifugation were cultured in vitro for 20 h with or without 10,50 or 100 ng/mL IL-1β,IL-6,IFN-γ,G-CSF or GM-CSF,or for 12 h with or without 1,10 or 50 ng/mL TNF-α.After incubation,cells were stained with annexin Ⅴ(AV)/propidium iodide(PI),and the proportions and absolute number of neutrophils undergoing different forms of cell death were determined by fluorescence microscopy combined with manual counting.Pyroptotic neutrophils were identified by cell morphology in conjunction with AV/PI staining.Flow cytometry with counting beads was employed to measure the proportions and number of AV/PI-stained Ly6g+neutrophils in different forms of cell death.Western blotting was employed to assess the cleavage and activation levels of cysteinyl aspartate-specific proteinase-3(caspase-3)and gasdermin E(GSDME).Results Treatment with IL-1β or IL-6 had no significant effect on the proportion or number of neutrophils undergoing spontaneous pyroptosis.After 12 h of treatment with TNF-α at 1,10,and 50 ng/mL,the proportions of pyroptotic neutrophils were(14.79±0.45)%,(19.99±3.02)%,and(20.66±1.99)%,respectively,higher than that[(10.22±1.12)%]in the untreated control(P=0.024,P<0.001,and P<0.001,respectively).Treatment with 10,50,and 100 ng/mL IFN-γ for 20 h reduced the proportion of pyroptotic neutrophils from(17.43±1.88)%to 12.00%(all P<0.001).G-CSF at 10,50,and 100 ng/mL reduced the proportion of pyroptotic cells to around 6.00%and greatly inhibited the cleavage of both caspase-3 and GSDME.After 20 h of treatment with 10,50,and 100 ng/mL GM-CSF,the proportions of pyroptotic neutrophils decreased to(7.52±0.53)%,(5.27±2.30)%,and(0.64±1.11)%,respectively.Conclusions Neither IL-1β nor IL-6 affects GSDME-mediated spontaneous pyroptosis in neutrophils.TNF-ɑ induces spontaneous pyroptosis in neutrophils,whereas IFN-γ,G-CSF,and GM-CSF demonstrate inhibitory effects.
Pyroptosis/drug effects*
;
Animals
;
Neutrophils/cytology*
;
Mice
;
Cytokines/pharmacology*
;
Interleukin-1beta/pharmacology*
;
Granulocyte-Macrophage Colony-Stimulating Factor/pharmacology*
;
Cells, Cultured
;
Granulocyte Colony-Stimulating Factor/pharmacology*
;
Tumor Necrosis Factor-alpha/pharmacology*
;
Interferon-gamma/pharmacology*
;
Interleukin-6/pharmacology*
8.Expression Changes of Hypoxia-Inducible Factor-1α in G-CSF Induced Hematopoietic Stem Cell Mobilization.
Hui-Xuan YANG ; Qiao-Chuan LI ; Li-Li WEI ; Yong-Rong LAI
Journal of Experimental Hematology 2023;31(1):221-226
OBJECTIVE:
To investigate the expression and its relative mechanism of hypoxia-inducible factor-1α(HIF-1α) in bone marrow(BM) of mice during G-CSF mobilization of hematopoietic stem cells (HSC) .
METHODS:
Flow cytometry was used to detect the proportion of Lin-Sca-1+ c-kit+ (LSK) cells in peripheral blood of C57BL/6J mice before and after G-CSF mobilization. And the expression of HIF-1α and osteocalcin (OCN) mRNA and protein were detected by RQ-PCR and immunohistochemistry. The number of osteoblasts in bone marrow specimens of mice was counted under the microscope.
RESULTS:
The proportion of LSK cells in peripheral blood began to increase at day 4 of G-CSF mobilization, and reached the peak at day 5, which was significantly higher than that of control group (P<0.05). There was no distinct difference in the expression of HIF-1α mRNA between bone marrow nucleated cells and osteoblasts of steady-state mice (P=0.073), while OCN mRNA was mainly expressed in osteoblasts, which was higher than that in bone marrow nucleated cells (P=0.034). After mobilization, the expression level of HIF-1α increased, but OCN decreased, and the number of endosteum osteoblasts decreased. The change of HIF-1α expression was later than that of OCN and was consistent with the proportion of LSK cells in peripheral blood.
CONCLUSION
The expression of HIF-1α in bone marrow was increased during the mobilization of HSC mediated by G-CSF, and one of the mechanisms may be related to the peripheral migration of HSC induced by osteoblasts inhibition.
Mice
;
Animals
;
Hematopoietic Stem Cell Mobilization
;
Granulocyte Colony-Stimulating Factor/pharmacology*
;
Hypoxia-Inducible Factor 1, alpha Subunit/metabolism*
;
Mice, Inbred C57BL
;
Bone Marrow Cells/metabolism*
;
Osteocalcin/metabolism*
;
RNA, Messenger/metabolism*
9.Relationship between promoting effect of Zuogui Pills on angiogenesis of reproductive organs and mobilization factors GM-CSF, SDF-1, and their receptors of EPCs in early-aging rats.
Qian-Qian SONG ; Heng DUAN ; Wan-Chun PENG ; Xiao-Jing WEI
China Journal of Chinese Materia Medica 2023;48(16):4467-4474
This study aimed to investigate the relationship between the promoting effect of Zuogui Pills on ovarian and vaginal angiogenesis in early-aging rats and mobilization factors granulocyte-macrophage colony-stimulating factor(GM-CSF), stromal cell-derived factor-1(SDF-1), and their receptors of endothelial progenitor cells(EPCs) and explore the mechanism of Zuogui Pills in improving reproductive hypofunction in early-aging rats. Ultra-high performance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS) was used to analyze the chemical components of the extract of Zuogui Pills. Forty 14-month-old female early-aging rats with estrous cycle disorder were randomly divided into a blank group, a conjugated estrogen group(conjugated estrogen suspension, 65 μg·kg~(-1)), and low-(11 g·kg~(-1)) and high-dose(33 g·kg~(-1)) Zuogui Pills groups, with 10 rats in each group. In addition, 10 4-month-old female rats were assigned to the youth control group. The rats in the blank group and the youth control group were treated with 20 g·kg~(-1) distilled water by gavage, while those in the groups with drug intervention were treated with corresponding drugs by gavage, once a day for 15 days. Enzyme-linked immunosorbent assay(ELISA) was used to detect the levels of SDF-1 and GM-CSF in the mobilization of EPCs in serum. Hematoxylin-eosin(HE) staining was used to observe the changes in the number of ovarian follicles at all levels and corpus luteum, the number of vaginal epithelial layers, the number of vaginal folds, and the blood vessels of ovarian and vaginal tissues in the groups with drug intervention. Western blot was used to detect the expression of ER, GM-CSFR, CXCR4, and CXCR7 proteins in ovarian and vaginal tissues. As revealed by the results, the blank group showed decreased number of corpus luteum, gro-wing follicles at all levels, and blood vessels(P<0.05), decreased thickness of vaginal mucosa, the number of epithelial layers, the number of vaginal folds, and the number of vessels in the lamina propria(P<0.05), reduced content of SDF-1 and GM-CSF in the peripheral blood(P<0.05), and down-regulated levels of ER, CXCR4, CXCR7, and GM-CSFR proteins in ovarian and vaginal tissues(P<0.05). The groups with drug intervention showed increased number of growing follicles at all levels, corpus luteum, and blood vessels(P<0.05), decreased number of atresia follicles(P<0.05), increased thickness of vaginal mucosa, the number of epithelial layers, the number of vaginal mucosal folds, and the number of blood vessels in the lamina propria(P<0.05), increased content of SDF-1 and GM-CSF in the peripheral blood(P<0.05), and up-regulated levels of ER, CXCR4, CXCR7, and GM-CSFR proteins in ovarian and vaginal tissues(P<0.05). This experiment suggests that Zuogui Pills may promote ovarian and vaginal angiogenesis and improve the reproductive function of early-aging rats by up-regulating the levels of mobilization factors SDF-1, GM-CSF, and their receptors of EPCs.
Rats
;
Female
;
Animals
;
Granulocyte-Macrophage Colony-Stimulating Factor
;
Estrogens, Conjugated (USP)
;
Tandem Mass Spectrometry
;
Aging
;
Genitalia
10.Efficacy and Safety of Plerixafor Combined with G-CSF for Autologous Peripheral Blood Hematopoietic Stem Cell Mobilization in Lymphoma Patients.
Fang-Shu GUAN ; Dong-Hua HE ; Yi LI ; Yi ZHANG ; Gao-Feng ZHENG ; Yuan-Yuan ZHU ; Jing-Song HE ; En-Fan ZHANG ; Zhen CAI ; Yi ZHAO
Journal of Experimental Hematology 2023;31(4):1056-1060
OBJECTIVE:
To investigate the efficacy and safety of plerixafor combined with granulocyte colony-stimulating factor (G-CSF) in mobilizing peripheral blood hematopoietic stem cells in patients with lymphoma.
METHODS:
The clinical data of lymphoma patients who received autologous hematopoietic stem cell mobilization using plerixafor combined with G-CSF from January 2019 to December 2021 were retrospectively analyzed. The patients received 3 kinds of mobilization regimens: front-line steady-state mobilization, preemptive intervention, and recuse mobilization. The acquisition success rate, excellent rate of collection, and incidence of treatment-related adverse reaction were counted. The influence of sex, age, disease remission status, bone marrow involvement at diagnosis, chemotherapy lines, number of chemotherapy, platelet count and number of CD34+ cells on the day before acquisition in peripheral blood on the collection results were analyzed to identify the risk factors associated with poor stem cell collection.
RESULTS:
A total of 43 patients with lymphoma were enrolled, including 7 cases who received front-line steady-state mobilization, 19 cases who received preemptive intervention, and 17 cases who received recuse mobilization. The overall acquisition success rate was 58.1% (25/43) after use of plerixafor combined with G-CSF, and acquisition success rate of front-line steady-state mobilization, preemptive intervention, and recuse mobilization was 100%, 57.9%(11/19), and 41.2%(7/17), respectively. The excellent rate of collection was 18.6%(8/43). A total of 15 patients experienced mild to moderate treatment-related adverse reactions. The number of CD34+ cells < 5 cells/μl in peripheral blood on the day before collection was an independent risk factor affecting stem cell collection.
CONCLUSIONS
Plerixafor combined with G-CSF is a safe and effective mobilization regimen for patients with lymphoma. The number of CD34+ cells in peripheral blood on the day before collection is an predictable index for the evaluation of stem cell collection.
Humans
;
Antigens, CD34/metabolism*
;
Granulocyte Colony-Stimulating Factor/therapeutic use*
;
Hematopoietic Stem Cell Mobilization/methods*
;
Hematopoietic Stem Cell Transplantation
;
Heterocyclic Compounds/therapeutic use*
;
Lymphoma/drug therapy*
;
Multiple Myeloma/drug therapy*
;
Retrospective Studies
;
Transplantation, Autologous

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