OBJECTIVE To analyze the efficacy and safety of luspatercept in the treatment of myelodysplastic syndromes （MDS） anemia， and provide reference for clinical medication. METHODS The literature related to luspatercept for MDS anemia in PubMed， Cochrane Library， Embase and Web of Science were searched by computer， and the search time was from the establishment of the database to January 2024. The quality of literature was evaluated after they were screened according to inclusion and exclusion criteria， the single-group rate meta-analysis and sensitivity analysis were performed by using RevMan 5.4 software， and the subgroup analysis was conducted. RESULTS A total of 756 patients in 9 articles were included in this study. The results of meta-analysis showed that the proportion of MDS patients who reached ≥8 weeks of red blood cell transfusion independence （RBC-TI） was 46% after using luspatercept [95%CI （0.28， 0.64）， P＜0.000 01]. The proportion of MDS patients whose hematological improvement in erythrocyte （HI-E） was 59% [95%CI （0.43， 0.74）， P＜0.000 01]. Among them， 5 articles reported that the proportion of MDS patients with grade 3-4 adverse reactions was 14% [95%CI （0.07， 0.22）， P＝0.000 2]， and the poor general condition， infection， blood and lymphatic system disease were the common adverse reactions. Subgroup analysis showed that the source of heterogeneity was the blood transfusion burden in the proportion of MDS patients with RBC-TI≥8 weeks， and the source of heterogeneity was the 0931-8356251。revised international prognostic scoring system （IPSS-R） risk grade， SF3B1 mutation status and blood transfusion burden in the proportion of MDS patients with HI-E. Sensitivity analysis showed that the results of this study were stable. CONCLUSIONS Luspatercept can significantly improve blood transfusion dependence， reduce blood transfusion burden and promote hematology improvement in MDS patients. But attention should be paid to the occurrence of grade 3-4 adverse events； adverse events such as poor general condition， infection， blood and lymphatic system diseases are more common.

Objective To investigate the impact of nucleophosmin 1-mutated(NPM1m)subtypes,variant allele frequency(VAF)and co-mutations on the survival outcomes of patients with acute myeloid leukemia(AML).Methods Clinical data,mutation status,and outcomes of 86 NPM1m-AML patients admitted in Department of Hematology,First Hospital of Lanzhou University between June 2017 and September 2024 were collected and retrospectively analyzed.Spearman correlation analysis,Kaplan-Meier curve analysis,Log-rank test,and Cox regression analysis was applied for correlation analysis,survival analysis,and factors affecting survival.Results The overall survival(OS)was significantly shorter in the Rares subtype of NPM1m than the ABD subtype(median OS:164 d vs 416 d,P=0.043).The VAF of NPM1m was positively correlated with the initial peripheral blood white blood cell count and lactate dehydrogenase(LDH)level(P<0.05).OS was reduced when VAF was≥0.37(median OS:164 d vs 730 d,P=0.003).The presence of myelodysplasia-related gene(MR)mutations was associated with a poorer prognosis when compared to the MR wild-type(median OS:45 d vs 395 d,P<0.001).The triple mutation of FMS-like tyrosine kinase 3-internal tandem duplication(FLT3-ITD)and DNMT3A also indicated a worse prognosis than the non-triple mutation(median OS:173 d vs 483 d,P=0.007).The presence of PTPN11 mutations was associated with improved OS(median OS:395 d vs 240 d,P=0.027),while the patients with coexistence of N/KRAS mutations trended toward better prognosis than the wild-type patients(median OS:662 d vs 189 d)though no statistical significance(P=0.070).Multivariate analysis revealed that LDH(HR=1.002,95%CI:1.000～1.003,P=0.005),NPM1m VAF(HR=2.415,95%CI:1.208～4.829,P=0.013),Rares subtype(HR=3.037,95%CI:1.134～8.136,P=0.027),and MR mutations(HR=5.283,95%CI:1.991～14.017,P<0.001)were independent risk factors associated with OS in the patients.Conclusion Molecular heterogeneity of NPM1m should be taken into account in prognostic stratification of NPM1m-AML.Factors including the Rares subtype,VAF≥0.37,coexistence of FLT3-ITD and DNMT3A mutations,and MR mutations are associated with poor prognosis of NPM1m-AML.The presence of PTPN11 mutations improves the prognosis,while the presence of N/KRAS mutations shows a trend toward better prognosis.LDH,NPM1m VAF,Rares subtype,and MR mutations are independent risk factors affecting OS of patients with NPM1m-AML.

Myelodysplastic syndrome(MDS)is a heterogeneous myeloid tumor that originates from hematopoietic stem cells(HSCs)and is associated with a high risk of progression to acute myeloid leukemia(AML).Studies have shown that 90%of patients with MDS have gene mutations,of whom approximately 25%have SF3B1 mutations.In patients with MDS carrying this mutation,the TGF-β pathway is upregu-lated,inducing cell cycle arrest and thereby leading to erythroid ineffective hematopoiesis and pathological hematopoiesis.Luspatercept can be used as a ligand trap to capture TGF-β ligands,inhibit SMAD2/3 pathway activation,downregulate TGF-β pathway,and promote ad-vanced red blood cell maturation.Currently,it has been approved by the Food and Drug Administration(FDA)for the treatment of anemia in patients with low-risk MDS,and studies have shown that the response rate is higher in patients with SF3B1 mutations.This article will re-view the current status of luspatercept in the treatment of SF3B1 mutation-related MDS;it will also analyze its effectiveness and safety and provide therapeutic strategies for clinical use.

Objective To study the therapeutic effects and mechanisms of Astragaloside-Ⅳ (ASG-Ⅳ) on diabetic cardiomyopathy (DCM) in the rat diabetic model. Methods Forty SD(Sprague-Dawley)healthy rats were used. The diabetic retinopathy rats model were induced by STZ, 45 mg/kg, 3d. Another 10 were injected the same amount of citrate buffer as a control group. Fasting blood glucose was measured with SureStep Plus detector 72 h later. The blood glucose of the diabetes model was ≥16.7 mmol/L. And 12 weeks later, DCM rats were divided into 4 groups randomly in the experiment, includes: DCM, ASG-Ⅳ-L (10 mg/kg), ASG-Ⅳ-M (30 mg/kg), ASG-Ⅳ-H (60 mg/kg)groups. After give dugs 4 weeks, the phosphokinase isoenzyme (CK-MB) and LDH level were tested, the cardiac hypertrophy was evaluated by HW/BW and LVW/BW. Activity of Na+-K+-ATPase and Ca2+-ATPase were determined in left ventricular tissues by ATPase ELISA Assay Kit. The content of FFA was measured and myocardial pathological examination was performed. Results Compared with the control group, blood and urine glucose were higher than experimental animal in diabetic model group, were significantly increased (P<0.05). LDH and Phosphokinase isoenzyme (CK-MB) level were significantly increased, the HWI and LVWI ratio were enhanced in DCM group (P<0.05). ASG-Ⅳ could reduce the ratio of HWI and LVWI, decrease the level of CK-MB and LDH, improve the pathomorphological changing of DCM rat model (P<0.05). Moreover, compared with DCM group, ASG-Ⅳ could restore the Na+-K+-ATPase and Ca2+-ATPase activity, reduced the content of FFA (P<0.05). Conclusion ASG-Ⅳ plays therapeutic effect on diabetic cardiomyopathy might via restore the Na+-K+-ATPase and Ca2+-ATPase activity, reduce the content of FFA, protect the myocardial energy metabolism in DCM.

A method was developed for simultaneous determination of Clozapine, Quetiapine and Risperidone in human serum by fully automated online solid phase extraction ( SPE )-high performance liquid chromatography. With Capcell MF Ph-1 column as SPE cartridge and Acclaim C18 as analytical column, high
selectivity could be achieved by this method according to the selective complementarity of the two columns. In the experiment, ACN-H2 O was used as the SPE mobile phase with a flow rate of 1 mL/min and ACN-10 mmol/L NH4 Ac was used as the analytical mobile phase with a flow rate of 1 mL/min. Serum samples were injected directly into the SPE column and the biological matrix was washed out with the loading solvent. By rotation of the switching valve, Clozapine, Quetiapine and Risperidone were eluted from the SPE cartridge in the back-flush mode and transferred to the analytical column by the chromatographic mobile phase. The whole time of the online SPE purification and chromatographic separation of the analytes was 18 min. Calibration curve of Clozapine with good linearity ( r=0 . 9996 ) was obtained in the range of 10-1800 μg/L in human serum, and the recoveries at low, medium and high concentration levels were 118. 4%, 105. 0% and 105 . 4%. Calibration curve of Quetiapine with good linearity ( r=0 . 9997 ) was obtained in the range of 3 . 6-640 μg/L in human serum, and the recoveries at low, medium and high concentration levels were 112. 8%, 101 . 1% and 101 . 5%. Calibration curve of Risperidone with good linearity ( r=0 . 9995 ) was obtained in the range of 0. 71-128 μg/L in human serum, and the recoveries at low, medium and high concentration levels were 100. 7%, 97. 2% and 98. 8%. In conclusion, the established automated online SPE-HPLC-UV method demonstrated good performance in terms of linearity, specificity, limits of quantification, and was successfully utilized to quantify Clozapine, Quetiapine and Risperidone in human serum.

Objective To analyze the effect of cyclosporin A(CsA), rapamycin(RPM) and macophenolic acid(MPA) on the co-stimulated lymphocytes, CD28 and CD40, and their production of Th1/Th2 cytokine, IL-2, IFN-γ, IL-4, IL-10 and IL-12. Methods The experimental groups were divided into ①mono-stimulating and co-stimulating groups: CD3 mAb mono-stimulating (group a),CD3 mAb+CD28 mAb co-stimulating (group b), CD3 mAb+CD28 mAb+CD40 L mAb co-stimulating(group c), CD3 mAb+CD28 mAb+CTLA4 mAb co-stimulating (group d). ②CsA groups: 300 ng/ml of CsA was added to group a, b, c and d. ③RPM groups: 300 ng/ml of RPM was added to group a,b, c and d. ④MPA groups:300 ng/ml of MPA was added to group a, b, c and d. The cytokine production was measured by ELISA.Results The co-stimulated CD28 and CD40 Th1/Th2 cytokines production of IFN-γ, IL-2, IL-4 and IL-10 were significantly increased. Compared with group a, IFN-γ increased from (248.91±11.20)ng/ml to (555.08±24.42)ng/ml and (548.19±33.06)ng/ml, IL-2 increased from (29.48±8.61)ng/ml to (1100.82±99.29)ng/ml and (842.23±29.31)ng/ml, IL-4 increased from (32.29±6.76)ng/ml to (116.02±15.03)ng/ml and (147.28±18.07)ng/ml, IL-10 increased from (147.01±10.47)ng/ml to (291.79±12.47)ng/ml and (302.52±35.18)ng/ml,respectively, P＜0. 01. Compared group b with group c, the Th1 cytokines production was decreased.IL-2 and IL-12 decreased (P＜0.05). The Th2 cytokine IL-4 production was increased (P＜0. 05).CTLA4 mAb and three other immunosuppressants, CsA, RPM and MPA, inhibited co-stimulated lymphocyte's both cytokines Th1 and Th2 production. The inhibitory effect of CsA on Th1/Th2 cytokine production was more significant than RPM and MPA did. The co-inhibitory effect of CTLA4 mAb and CsA was observed as well. The increased co-stimulated CD28 and CD40 IL-12 production could be suppressed by MPA. CsA and RPM had no inhibitory effect on the IL-12 production.Conclusions CD28/CD40 co-stimulatory pathway plays the key role in lymphocyte activation and Th1/Th2 cytokine production. CsA, RPM and MPA can inhibit co-stimulated lymphocyte's Th1 and Th2 cytokine production. CsA and CTLA4 mAb have co-inhibitory effect on co-stimulated lymphocyte's Th1/Th2 cytokines production. CD40 L mAb decreases the Th1 cytokines production(including IL-12) and increases the Th2 (mainly IL-4) production, which may be the mechanism of its longevity effect on allograft.

To develop the methodology to purify and culture expand mesenchymal stem cells (MSC) from human bone marrow and investigate the optimal system for MSC differentiation into chondrocytes mononuclear cells were harvested by gradient centrifugation on Percoll at density of 1 073g/ml and seeded in low glucose DMEM containing 10% fetal calf serum. The purity of cells was evaluated by flow cytometric technique. MSC of 2 passages thereafter were absorbed into a biomaterial of gelatin sponge and induced to differentiate in to chondrocytes for one week under the influence of TGF ? and other inductive agents. The feature of chondrocytes in engineered tissues was identified by toluidine blue staining at various time points. The results showed that human mesenchymal stem cells culture expanded were positive for CD166, CD29, and CD44, but negative for CD34, CD45, and HLA DR. Furthermore, when treated cells absorbed into the biomaterial were implanted subcutaneously into BALB c nude mice, they formed cartilage like tissues after 4 weeks.Our conclusions is that cultured marrow MSC have unique biological features and the capacity to differentiate into chondrocytes in vivo , so they are useful for cartilage engineering by serving as the seed cells.

Objective To study the therapeutic window of rapamycin(RPM) concentration in primary recipients of renal transplantation. Methods An open label, multi center study was performed. One hundred primary renal allograft recipients with cadaveric donors were enrolled from 4 transplantation centers in China. The immunosuppressive regimen was triple therapy,i.e.RPM combined with CsA and steroid. A loading dose of RPM 6 mg/d was administered within 48 hours after transplantation, then a maintaining dose of 2 mg/d was administered. The whole blood concentration of RPM was measured by HPLC method. Results The whole blood concentration of RPM in this group was (6.65?2.75)ng/ml, the 10th and 90th percentile for RPM concentration was 3 2 ng/ml and 10 26 ng/ml,respectively.9 5%(8/84)patients suffered from acute rejection during the 6 month period after transplantation in this study, and the concentration of RPM in these was lower than that in non rejection patients(P=0.001). Hyperlipidemia and liver dysfunction were the most frequently adverse events, and RPM concentration was significantly associated with the concentration of triglyceride. Conclusions 4～8 ng/ml is a suitable level for RPM concentration. Regular drug monitoring and reasonable dose modulation may increase the validity and security of RPM.

Objective To observe the dynamic change of cytokines and effect of different stimulating combinations on dominant regulation, and describe the microenviroment character of local immune response. Methods PBMCs were separated from peripheral blood of healthy donors with density gradient centrifugation. The expression levels of cytokine genes under different stimulating combinations, including IL-2, IFN-?, IL-12, IL-4, IL-10 and TGF-?1, were detected by ELISA and RT-PCR. Results ELISA results indicated that secretion level of IL-2, IL-4, IL-12 and IFN-? were inhibited by IL-10. There were significant differences in IL-4 and IL-12 between control group and stimulating groups under the stimulation by anti-CD3 monoclonal antibody (McAb) or anti-CD3 McAb + anti-CD28 McAb combination. Significant differences in IL-2 only appeared in anti-CD3 antibody stimulating group. The concentrations of IFN-? were decreased moderately. At the mean time, IL-10 significantly promoted the secretion of TGF-?1 under the anti-CD3 McAb stimulation. There were similar results on the genes level for the studied cytokines. Conclusion The induction and maintenance of dominant regulation were dependent mainly on IL-10, which inhibits the overexpression of IL-4 and IL-12. It is the vital step for dominant regulation to avoid the mono-polarization development at the beginning of immune response.