Objective:To investigate GPRC5D expression on myeloma cells in newly diagnosed multiple myeloma (NDMM) patients and evaluate its prognostic significance.Methods:This study retrospectively analyzed the clinical data of 65 patients with NDMM treated at the Affiliated Hospital of Xuzhou Medical University from April 2023 to April 2024. The expression of GPRC5D on the surface of myeloma cells in all patients was detected with flow cytometry before induction therapy, and patients were stratified into high and low GPRC5D expression groups based on the median GPRC5D positivity rate. Clinical characteristics, immune status, treatment response after induction therapy, and prognosis were compared between the two groups.Results:The median positive rate of GPRC5D in the plasma cells of 65 patients with NDMM was 32.68%. Based on this threshold, patients were categorized into the high (33 cases, GPRC5D positive rate ≥ 32.68%) and low (32 cases, GPRC5D positive rate <32.68%) GPRC5D expression groups. Compared with the low GPRC5D expression group, the high GPRC5D expression group demonstrated a higher proportion of 1q21 gain (78.8% vs 43.8%, P=0.004), a higher incidence of immunoparesis involving ≥2 uninvolved immunoglobulins (87.9% vs 62.5%, P=0.018), and severe immunoparesis (59.4% vs 33.3%, P=0.046). Further, CD16 +CD56 + cell levels were lower in the high GPRC5D expression group [ (16.60±8.70) % vs (27.78±15.78) %, P=0.005]. No significant difference was observed in the overall response rate between the high and low GPRC5D expression groups (78.8% vs 93.8%, P=0.165). However, the high GPRC5D expression group exhibited a significantly lower rate of achieving very good partial remission or better (42.4% vs 78.2%, P=0.003) and a lower MRD negativity rate (30.0% vs 68.8%, P=0.002). Compared with the low GPRC5D expression group, patients with high expression demonstrated a significantly shorter median progression-free survival (11.2 months vs not reached, P=0.002), whereas the median overall survival was not reached in either group, with no statistically significant difference ( P=0.069) . Conclusions:The GPRC5D positivity rate in the plasma cells of patients with NDMM is associated with 1q21 gain and immune status. High GPRC5D expression at diagnosis may predict poor response to induction therapy and an unfavorable prognosis.

Objective:To investigate the combined therapeutic role of the AKT inhibitor MK2206 and Ruxolitinib in treating Myeloproliferative Neoplasms (MPN) driven by a calreticulin (CALR) gene mutation.Methods:① Murine bone marrow c-kit + cells were isolated by sacrificing mice and harvesting bone marrow from the femur, tibia, and ilium for subsequent c-kit + cell sorting. ② A CALR transplantation mouse model was established. GFP-tagged retroviral vectors containing either the CALR gene mutation or the migR1 control were constructed, packaged in Platinum-E cells, and used to transduce murine bone marrow c-kit + cells. These transduced cells were then transplanted into lethally irradiated female recipient mice via tail vein injection. ③ Following successful engraftment, the mice were randomly assigned to four treatment groups for intragastric administration. Complete blood counts were monitored periodically, and the spleen size and weight of transplanted mice were measured. ④ Flow cytometry was used to quantify the proportions of GFP + tumor cells, megakaryocytic lineage cells, and hematopoietic stem cells in both splenic and bone marrow tissues. Histopathological examination was performed to evaluate the degree of tumor cell infiltration in these organs. Results:① Following gavage treatment, peripheral blood platelet (PLT) and white blood cell counts were significantly lower in the combined AKT inhibitor MK2206 and Ruxolitinib group compared to the MK2206, Ruxolitinib, and control groups ( P<0.05). ② In comparison with the MK2206 and Ruxolitinib monotherapy groups, the combination therapy group exhibited a significant reduction in spleen weight and a marked improvement in splenomegaly at 30 weeks post-transplantation ( P<0.05). ③ After four weeks of continuous treatment, combined administration resulted in a significant decrease in the proportion of megakaryocytic lineage cells and GFP + tumor cells in the bone marrow and spleen ( P<0.05). Additionally, the proportion of hematopoietic stem cells in the bone marrow was also significantly reduced ( P<0.05). ④ Histopathological analysis (H&E staining) of bone marrow and spleen tissues confirmed that the combined regimen decreased both tumor cell infiltration and the proportion of abnormal megakaryocytes in these organs. Conclusion:The combination of AKT inhibitor MK2206 and Ruxolitinib is effective at significantly ameliorating disease symptoms and reducing tumor infiltration in vivo in mice with a myeloproliferative tumor transplantation driven by a CALR gene mutation.

Objective:To investigate the relationship between retinoblastoma binding protein 1 (RB1) gene deletion and the prognosis of multiple myeloma (MM) patients, and the possible effect of renal insufficiency on it.Methods:A retrospective cohort study was conducted. The clinical data and follow-up information of MM patients who were treated in the Second Affiliated Hospital of Xuzhou Medical University and the Affiliated Hospital of Xuzhou Medical University from December 2020 to November 2023 were collected. According to the presence of RB1 gene deletion in bone marrow samples detected by fluorescence in situ hybridization (FISH), the patients were divided into the RB1 gene deletion group and the RB1 gene non-deletion group, and the clinicopathological characteristics and hematological index levels were compared between the two groups. Renal insufficiency was determined by renal function assessment indicator serum creatinine (Scr) >177 μmol/L. The Spearman test was used to analyze the relationship between the number of RB1 gene deletion positive cells and levels of Scr, hemoglobin and serum calcium in MM patients. The Kaplan-Meier method was used to analyze progression-free survival (PFS), and the Cox proportional hazards model was used to determine the influencing factors of PFS in all MM patients and RB1 gene deletion and non-deletion MM patients.Results:A total of 75 MM patients were enrolled, of whom 24 (32.0%) had RB1 gene deletion. There were no significant differences in gender, age ≥65 years old, bone destruction and lactate dehydrogenase level between the RB1 gene deletion and non-deletion groups (all P > 0.05). There were significant differences in the distributions of patients in each stage of MM International Staging System (ISS) and revised International Staging System (R-ISS) between the two groups, as well as in hemoglobin, serum calcium, Scr, β 2-microglobulin, serum albumin levels, and the proportion of bone marrow plasma cells (all P < 0.05). The number of RB1 gene deletion positive cells was positively correlated with Scr level ( r = 0.863, P = 0.016), but not with hemoglobin and serum calcium levels (both P > 0.05). The PFS of the RB1 gene non-deletion group was better than that of the RB1 gene deletion group (1-year PFS rate: 83.5% vs. 71.7%, 2-year PFS rate: 56.3% vs. 26.3%), and the difference was statistically significant ( P = 0.012). PFS in the non-renal insufficiency group was better than that in the renal insufficiency group (1-year PFS rate: 85.6% vs. 61.9%, 2-year PFS rate: 58.0% vs. 13.5%), and the difference was statistically significant ( P = 0.001). The PFS of patients without renal insufficiency in both the RB1 gene deletion and non-deletion groups was better than that in patients with renal insufficiency, and the differences were statistically significant (both P < 0.05). Multivariate Cox regression analysis showed that ISS stage Ⅲ was an independent risk factor for poor PFS in MM patients (stage Ⅲ vs. stage Ⅰ, HR = 11.317, 95% CI: 1.220-104.979, P = 0.033). Multivariate Cox regression analysis in RB1 gene deletion and non-deletion groups showed that ISS stage Ⅲ (stage Ⅲ vs. stageⅠ, HR = 4.166, 95% CI: 1.419-12.225, P = 0.009), R-ISS stage Ⅲ (stage Ⅲ vs. stage Ⅰ, HR = 3.800, 95% CI: 1.005-14.367, P = 0.049), serum calcium > 2.52 mmol/L (> 2.52 mmol/L vs. ≤2.52 mmol/L, HR = 2.398, 95% CI: 1.037-5.546, P = 0.041) and renal insufficiency (yes vs. no, HR = 2.363, 95% CI: 1.021-5.472, P = 0.045) were independent risk factors for poor PFS in RB1 gene non-deletion MM patients, and serum calcium >2.52 mmol/L (>2.52 mmol/L vs. ≤ 2.52 mmol/L, HR = 3.673, 95% CI: 1.160-11.627, P = 0.027) and renal insufficiency (yes vs. no, HR = 3.985, 95% CI: 1.220-13.016, P = 0.022) were independent risk factors for poor PFS in RB1 gene deletion MM patients. Conclusions:The PFS of MM patients with RB1 gene deletion is worse than that of patients without RB1 gene deletion, RB1 gene deletion may be related to renal insufficiency in MM patients, and the prognosis of MM patients with RB1 gene deletion and renal insufficiency may be worse.

Targeting T-cell is a strategy to control allogeneic response disorders, such as acute graft-versus-host disease (GVHD) which is an important cause of therapy-failure after allogeneic hematopoietic cell transplants. Free fatty acid receptor-4 (FFAR4) is a regulator of obesity but its role in T-cell and allogeneic reactions is unknown. Here, we found knockout of Ffar4 in donor T-cells in a mouse allograft model increased acute GVHD whereas the natural FFAR4 ligands and the synthetic FFAR4 agonists decreased it. FFAR4 agonist-mediated anti-acute GVHD effects depended on FFAR4-expression in donor T-cells. The FFAR4 agonist CpdA suppressed donor T-cell-mediated alloreaction by activating an aryl hydrocarbon receptor (AhR) pathway. CpdA recruited β-Arrestin2 to FFAR4 which facilitated nuclear translocation of AhR and upregulation of IL-22. The CpdA-mediated anti-acute GVHD effect was absent in mice receiving Ahr-knockout or Il22-knockout T-cells. Recipient-expressing Ffar4 was also important for the anti-acute GVHD effect of CpdA which inhibited activation of antigen presenting cells. Importantly, CpdA decreased acute GVHD in obese mice, an effect also depended on Ffar4-expression in donor T-cells and recipients. Our study shows the immunoregulatory effect of FFAR4 in T-cell, and targeting FFAR4 might be a relative option for controlling allogeneic reactions in obese patients.

OBJECTIVES: To evaluate the photothermal and antibacterial activities of polydopamine-modified phycocyanin nanoparticles (PDA@PC NPs) and their capacity for promoting wound healing. METHODS: PDA@PC NPs were synthesized from phycocyanin (C-PC) and dopamine hydrochloride using a one-pot method. The photothermal activity of the nanoparticles was assessed in vitro by 808 nm laser irradiation, their biocompatibility was evaluated using CCK-8 assay, and their photothermal antibacterial activity by plate colony counting. In adult male BALB/c mice, two symmetrical full-thickness skin wounds (1.0 cm ×1.0 cm) were created on both sides of the spine, and 200 μL of Staphylococcus aureus suspension was inoculated into the wounds. The mice were divided into control group, PDA@PC NPs group, and PDA@PC NPs with laser irradiation group, and wound healing rates and histomorphological changes in the wound tissues were evaluated on days 0, 7 and 14 after modeling. RESULTS: The synthesized PDA@PC NPs exhibited no obvious cytotoxicity up to a concentration of 500 μg/mL and showed strong photothermal and antibacterial activities in response to 808 nm laser irradiation. In the mouse models, the size of the infected skin wounds showed substantial reduction at 7 and 14 days in PDA@PC NPs group and PDA@PC NPs with laser irradiation group, and the mean wound healing rate was faster in the latter group. HE staining and Masson's trichrome staining revealed extensive granulation tissue formation and collagen deposition on the wound surfaces in both of the treatment groups, and these changes were more obvious in the PDA@PC NPs with laser irradiation group. CONCLUSIONS PDA@PC NPs possess excellent photothermal and antibacterial activities and can effectively promote wound healing in mice.
Animals ; Indoles/chemistry* ; Wound Healing/drug effects* ; Mice ; Mice, Inbred BALB C ; Male ; Nanoparticles ; Polymers/chemistry* ; Phycocyanin/chemistry* ; Skin/injuries* ; Staphylococcus aureus/drug effects* ; Anti-Bacterial Agents/pharmacology*

Objective:To investigate the efficacy and safety of avatrombopag in promoting hematopoietic reconstitution after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Method:A retrospective analysis was conducted on 60 recipients with hematological malignancies who underwent allo-HSCT at the Affiliated Hospital of Xuzhou Medical University from January 2022 to August 2023. Recipients with hepatic or renal insufficiency before conditioning, those who received other thrombopoietic agents after allo-HSCT, those with severe respiratory or circulatory system diseases, and those with a history of thromboembolic events were excluded. Among them, 30 recipients who received avatrombopag within 14 days post-transplantation were assigned to the avatrombopag group, while the remaining 30 recipients who did not receive any thrombopoietic agents served as the control group. Clinical characteristics, hematopoietic stem cell engraftment, bone marrow proliferation, transfusion requirements, transplant-related complications, and laboratory adverse events were compared between the two groups.Result:The median platelet engraftment time in the avatrombopag group was 13 days (range: 9~25 days), and the neutrophil engraftment time was 13 days (range: 11~21 days). In the control group, he platelet engraftment time was 15 days (range: 10~51 days), and neutrophil engraftment time was 14 days (range: 10~30 days). The difference in platelet engraftment time between the two groups was statistically significant ( P=0.039). Bone marrow analysis on day 28 post-transplant showed that the proportion of recipients with active bone marrow hyperplasia was 96.7% in the avatrombopag group and 73.3% in the control group ( P=0.030); the median number of megakaryocytes was 30 vs. 6, respectively ( P<0.001); and the proportion of mature platelet-producing megakaryocytes was 44% vs. 26.3% ( P<0.001). Regarding transfusion requirements, the median platelet transfusion volume within 28 days post-transplantation was 4.5 U (range: 2~16 U) in the avatrombopag group and 6.5 U (range: 3~32 U) in the control group ( P=0.007). The time to achieve platelet transfusion independence was 13 days (range: 8~25 days) in the avatrombopag group and 14 days (range: 10~36 days) in the control group ( P=0.026). The median red blood cell transfusion volume in both groups was 4 U, with no significant difference ( P=0.354). Medication adherence in the avatrombopag group was 100%. There were no statistically significant differences between the two groups in terms of incidence of post-transplant infections (70% vs. 83.3%), bleeding (50% vs. 60%), graft-versus-host disease (GVHD) (30% vs. 40%), or abnormal laboratory tests (86.7% vs. 90%) (all P>0.05). Conclusion:The use of avatrombopag after allo-HSCT in patients with hematologic malignancies can promote bone marrow hematopoiesis and platelet engraftment, reduce platelet transfusion volume, and shorten the duration of platelet transfusion dependence. Avatrombopag is well tolerated, and no serious adverse reactions were observed during treatment.

Objective:To investigate GPRC5D expression on myeloma cells in newly diagnosed multiple myeloma (NDMM) patients and evaluate its prognostic significance.Methods:This study retrospectively analyzed the clinical data of 65 patients with NDMM treated at the Affiliated Hospital of Xuzhou Medical University from April 2023 to April 2024. The expression of GPRC5D on the surface of myeloma cells in all patients was detected with flow cytometry before induction therapy, and patients were stratified into high and low GPRC5D expression groups based on the median GPRC5D positivity rate. Clinical characteristics, immune status, treatment response after induction therapy, and prognosis were compared between the two groups.Results:The median positive rate of GPRC5D in the plasma cells of 65 patients with NDMM was 32.68%. Based on this threshold, patients were categorized into the high (33 cases, GPRC5D positive rate ≥ 32.68%) and low (32 cases, GPRC5D positive rate <32.68%) GPRC5D expression groups. Compared with the low GPRC5D expression group, the high GPRC5D expression group demonstrated a higher proportion of 1q21 gain (78.8% vs 43.8%, P=0.004), a higher incidence of immunoparesis involving ≥2 uninvolved immunoglobulins (87.9% vs 62.5%, P=0.018), and severe immunoparesis (59.4% vs 33.3%, P=0.046). Further, CD16 +CD56 + cell levels were lower in the high GPRC5D expression group [ (16.60±8.70) % vs (27.78±15.78) %, P=0.005]. No significant difference was observed in the overall response rate between the high and low GPRC5D expression groups (78.8% vs 93.8%, P=0.165). However, the high GPRC5D expression group exhibited a significantly lower rate of achieving very good partial remission or better (42.4% vs 78.2%, P=0.003) and a lower MRD negativity rate (30.0% vs 68.8%, P=0.002). Compared with the low GPRC5D expression group, patients with high expression demonstrated a significantly shorter median progression-free survival (11.2 months vs not reached, P=0.002), whereas the median overall survival was not reached in either group, with no statistically significant difference ( P=0.069) . Conclusions:The GPRC5D positivity rate in the plasma cells of patients with NDMM is associated with 1q21 gain and immune status. High GPRC5D expression at diagnosis may predict poor response to induction therapy and an unfavorable prognosis.

Objective:To investigate the combined therapeutic role of the AKT inhibitor MK2206 and Ruxolitinib in treating Myeloproliferative Neoplasms (MPN) driven by a calreticulin (CALR) gene mutation.Methods:① Murine bone marrow c-kit + cells were isolated by sacrificing mice and harvesting bone marrow from the femur, tibia, and ilium for subsequent c-kit + cell sorting. ② A CALR transplantation mouse model was established. GFP-tagged retroviral vectors containing either the CALR gene mutation or the migR1 control were constructed, packaged in Platinum-E cells, and used to transduce murine bone marrow c-kit + cells. These transduced cells were then transplanted into lethally irradiated female recipient mice via tail vein injection. ③ Following successful engraftment, the mice were randomly assigned to four treatment groups for intragastric administration. Complete blood counts were monitored periodically, and the spleen size and weight of transplanted mice were measured. ④ Flow cytometry was used to quantify the proportions of GFP + tumor cells, megakaryocytic lineage cells, and hematopoietic stem cells in both splenic and bone marrow tissues. Histopathological examination was performed to evaluate the degree of tumor cell infiltration in these organs. Results:① Following gavage treatment, peripheral blood platelet (PLT) and white blood cell counts were significantly lower in the combined AKT inhibitor MK2206 and Ruxolitinib group compared to the MK2206, Ruxolitinib, and control groups ( P<0.05). ② In comparison with the MK2206 and Ruxolitinib monotherapy groups, the combination therapy group exhibited a significant reduction in spleen weight and a marked improvement in splenomegaly at 30 weeks post-transplantation ( P<0.05). ③ After four weeks of continuous treatment, combined administration resulted in a significant decrease in the proportion of megakaryocytic lineage cells and GFP + tumor cells in the bone marrow and spleen ( P<0.05). Additionally, the proportion of hematopoietic stem cells in the bone marrow was also significantly reduced ( P<0.05). ④ Histopathological analysis (H&E staining) of bone marrow and spleen tissues confirmed that the combined regimen decreased both tumor cell infiltration and the proportion of abnormal megakaryocytes in these organs. Conclusion:The combination of AKT inhibitor MK2206 and Ruxolitinib is effective at significantly ameliorating disease symptoms and reducing tumor infiltration in vivo in mice with a myeloproliferative tumor transplantation driven by a CALR gene mutation.

Objective:To investigate the relationship between retinoblastoma binding protein 1 (RB1) gene deletion and the prognosis of multiple myeloma (MM) patients, and the possible effect of renal insufficiency on it.Methods:A retrospective cohort study was conducted. The clinical data and follow-up information of MM patients who were treated in the Second Affiliated Hospital of Xuzhou Medical University and the Affiliated Hospital of Xuzhou Medical University from December 2020 to November 2023 were collected. According to the presence of RB1 gene deletion in bone marrow samples detected by fluorescence in situ hybridization (FISH), the patients were divided into the RB1 gene deletion group and the RB1 gene non-deletion group, and the clinicopathological characteristics and hematological index levels were compared between the two groups. Renal insufficiency was determined by renal function assessment indicator serum creatinine (Scr) >177 μmol/L. The Spearman test was used to analyze the relationship between the number of RB1 gene deletion positive cells and levels of Scr, hemoglobin and serum calcium in MM patients. The Kaplan-Meier method was used to analyze progression-free survival (PFS), and the Cox proportional hazards model was used to determine the influencing factors of PFS in all MM patients and RB1 gene deletion and non-deletion MM patients.Results:A total of 75 MM patients were enrolled, of whom 24 (32.0%) had RB1 gene deletion. There were no significant differences in gender, age ≥65 years old, bone destruction and lactate dehydrogenase level between the RB1 gene deletion and non-deletion groups (all P > 0.05). There were significant differences in the distributions of patients in each stage of MM International Staging System (ISS) and revised International Staging System (R-ISS) between the two groups, as well as in hemoglobin, serum calcium, Scr, β 2-microglobulin, serum albumin levels, and the proportion of bone marrow plasma cells (all P < 0.05). The number of RB1 gene deletion positive cells was positively correlated with Scr level ( r = 0.863, P = 0.016), but not with hemoglobin and serum calcium levels (both P > 0.05). The PFS of the RB1 gene non-deletion group was better than that of the RB1 gene deletion group (1-year PFS rate: 83.5% vs. 71.7%, 2-year PFS rate: 56.3% vs. 26.3%), and the difference was statistically significant ( P = 0.012). PFS in the non-renal insufficiency group was better than that in the renal insufficiency group (1-year PFS rate: 85.6% vs. 61.9%, 2-year PFS rate: 58.0% vs. 13.5%), and the difference was statistically significant ( P = 0.001). The PFS of patients without renal insufficiency in both the RB1 gene deletion and non-deletion groups was better than that in patients with renal insufficiency, and the differences were statistically significant (both P < 0.05). Multivariate Cox regression analysis showed that ISS stage Ⅲ was an independent risk factor for poor PFS in MM patients (stage Ⅲ vs. stage Ⅰ, HR = 11.317, 95% CI: 1.220-104.979, P = 0.033). Multivariate Cox regression analysis in RB1 gene deletion and non-deletion groups showed that ISS stage Ⅲ (stage Ⅲ vs. stageⅠ, HR = 4.166, 95% CI: 1.419-12.225, P = 0.009), R-ISS stage Ⅲ (stage Ⅲ vs. stage Ⅰ, HR = 3.800, 95% CI: 1.005-14.367, P = 0.049), serum calcium > 2.52 mmol/L (> 2.52 mmol/L vs. ≤2.52 mmol/L, HR = 2.398, 95% CI: 1.037-5.546, P = 0.041) and renal insufficiency (yes vs. no, HR = 2.363, 95% CI: 1.021-5.472, P = 0.045) were independent risk factors for poor PFS in RB1 gene non-deletion MM patients, and serum calcium >2.52 mmol/L (>2.52 mmol/L vs. ≤ 2.52 mmol/L, HR = 3.673, 95% CI: 1.160-11.627, P = 0.027) and renal insufficiency (yes vs. no, HR = 3.985, 95% CI: 1.220-13.016, P = 0.022) were independent risk factors for poor PFS in RB1 gene deletion MM patients. Conclusions:The PFS of MM patients with RB1 gene deletion is worse than that of patients without RB1 gene deletion, RB1 gene deletion may be related to renal insufficiency in MM patients, and the prognosis of MM patients with RB1 gene deletion and renal insufficiency may be worse.