Objective:To evaluate the efficacy of purified donor CD34 positive hematopoietic stem cell (CD34 + cell) infusion in patients with poor graft function (PGF) after allogeneic hematopoietic stem cell transplantation (allo-HSCT) . Methods:The clinical data of 25 patients with PGF who underwent donor CD34 + cell sorting and infusion at the Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences between September 2019 and March 2023, were retrospectively analyzed. The cohort included 19 haploidentical and 6 HLA-matched cases. CD34 + cells were purified using immunomagnetic beads for therapeutic infusion. The purification efficiency was evaluated based on the purity and recovery rate of CD34 + cells. Clinical outcomes were assessed by hematopoietic recovery, overall survival (OS) rate, and the incidence of graft-versus-host disease (GVHD) . Results:The median total number of CD34 + cells was 2.64 (0.82-6.53) × 10 8 before purification and 2.22 (0.48-5.68) ×10 8 after purification, with a median recovery rate of 78.37% (58.48%-115.72%) . After infusion of purified CD34 + cells, 8 of 10 patients (80.0%) with poor neutrophil engraftment achieved recovery (absolute neutrophil count ≥ 0.5×10 9/L) , with a median time to recovery of 21 (10-40) days. And 15 of 21 patients (71.4%) with poor platelet engraftment achieved recovery (platelet count ≥ 20×10 9/L) , with a median time to recovery of 15 (13-38) days. Only 3 patients (12.0%) developed GVHD after the infusion of purified CD34 + cells, including 2 cases of grade I acute GVHD and 1 case of limited chronic GVHD. With a median follow-up of 14.47 (0.23-41.63) months, the overall OS rate after CD34 + cell infusion was (65.63± 8.28) %. Seventeen patients survived, with a median survival time of 19.07 (0.23-41.63) months. Conclusion:Purification of CD34 + cells using immunomagnetic beads is effective, and the infusion of these purified donor CD34 + cells can safely and effectively improve PGF after allo-HSCT.

Objective:To evaluate the efficacy of purified donor CD34 positive hematopoietic stem cell (CD34 + cell) infusion in patients with poor graft function (PGF) after allogeneic hematopoietic stem cell transplantation (allo-HSCT) . Methods:The clinical data of 25 patients with PGF who underwent donor CD34 + cell sorting and infusion at the Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences between September 2019 and March 2023, were retrospectively analyzed. The cohort included 19 haploidentical and 6 HLA-matched cases. CD34 + cells were purified using immunomagnetic beads for therapeutic infusion. The purification efficiency was evaluated based on the purity and recovery rate of CD34 + cells. Clinical outcomes were assessed by hematopoietic recovery, overall survival (OS) rate, and the incidence of graft-versus-host disease (GVHD) . Results:The median total number of CD34 + cells was 2.64 (0.82-6.53) × 10 8 before purification and 2.22 (0.48-5.68) ×10 8 after purification, with a median recovery rate of 78.37% (58.48%-115.72%) . After infusion of purified CD34 + cells, 8 of 10 patients (80.0%) with poor neutrophil engraftment achieved recovery (absolute neutrophil count ≥ 0.5×10 9/L) , with a median time to recovery of 21 (10-40) days. And 15 of 21 patients (71.4%) with poor platelet engraftment achieved recovery (platelet count ≥ 20×10 9/L) , with a median time to recovery of 15 (13-38) days. Only 3 patients (12.0%) developed GVHD after the infusion of purified CD34 + cells, including 2 cases of grade I acute GVHD and 1 case of limited chronic GVHD. With a median follow-up of 14.47 (0.23-41.63) months, the overall OS rate after CD34 + cell infusion was (65.63± 8.28) %. Seventeen patients survived, with a median survival time of 19.07 (0.23-41.63) months. Conclusion:Purification of CD34 + cells using immunomagnetic beads is effective, and the infusion of these purified donor CD34 + cells can safely and effectively improve PGF after allo-HSCT.

【Objective】 To explore the feasibility of blood transfusion compatibility testing for multiple myeloma(MM) patients treated with anti-CD38 monoclonal antibody daratumumab (DARA) after DARA-Fab fragment blocking, and to evaluate the transfusion efficacy by comparing with dithiothreitol(DTT) method. 【Methods】 After DARA was prepared into DARA-Fab fragments using PierceFab preparation kit, the neutralization effects of different volumes (5, 10, 15, 30 μL) on screening cells and panel cells were confirmed. DARA-Fab fragments and screening cells with specific antigens and corresponding monoclonal antibody reagents were used as the experimental group and the control group with the same volume of saline for incubating and centrifugin.Twenty MM patients treated with DARA were selected for cross-matching with DARA-Fab and DTT respectively, and the laboratory indexes before and after transfusion were statistically analyzed, and the two blood matching methods were compared. 【Results】 After incubating and centrifuging, the results of DARA-Fab fragments(15, 30 μL) with screening cells and serum mixed with DARA were negative, while those of DARA-Fab(5, 10 μL) were positive. 15μL DARA-Fab treated antibody identification cells (2, 3, 4, 5, 7, 9, 11) were negative, antibody identification cells (1, 6, 8, 10, 12) were negative after 30 μL DARA-Fab fragments treatment; the results of MNS, Duffy, Kidd, Kell, Lewis, Rh blood group system of the experimental group were consistent with those of the control group; the hemoglobin (Hb) (g/L) of 20 patients after infusion of RBC (73.90±1.90) was significantly higher than that before transfusion (63.60±1.58), P<0.01. There was no significant difference in total bilirubin(TBil)(μmol/L)(16.25±3.54 vs 17.87±3.57), direct bilirubin(DBIL)(μmol/L)(6.31±2.32 vs 7.10±2.80)and indirect bilirubin(I-Bil)(9.94±1.38 vs 10.77±1.22) before and after infusion(P>0.05).And no statistical difference was noticed in Hb (10.75±1.04 vs 10.30±0.98), TBil (3.31±1.47 vs 3.31±0.55), DBIL(2.76±1.24 vs 2.60±0.83), and I-Bil(1.97±0.40 vs 2.82±0.53) between the DTT treatment method and the DARA Fab fragment treatment before and after transfusion(P>0.05). 【Conclusion】 DARA-Fab can remove the interference of RBC on cross matching by blocking CD38 antigen. This method has no effect on the antigens of common RBC blood group systems, and shows significant blood transfusion efficacy as that of DTT method.

Guided by the emphasis on learning process, the educational reform has designed a "trinity" comprehensive evaluation system (quantitative clinical practice, in-class medical record analysis, and staged comprehensive written test) as the formative evaluation of the course. Through this assessment system, students' self-learning potential is stimulated, clinical skills practice is strengthened, and "taking exams to promote learning and taking exams to promote teaching" is realized. In the practice of teaching reform, it has been found that compared with the conventional teaching class, the students in the teaching reform class have higher participation and are more satisfied with the process assessment of the "trinity" comprehensive evaluation system.

Objective:To study the effect of the siderophore virulence gene entB on the virulence of carbapenem-resistant Klebsiella pneumonia (CRKP). Methods:CRKP-27 was selected as the experimental strain from 30 CRKP strains collected from the First Affiliated Hospital of Kunming Medical University. The knockdown strain (Δ entB) and complementing strain (C-Δ entB) were constructed by the clustered regularly interspaced short palindromic repeat-Cas9 technology, and verified by polymerase chain reaction (PCR). In order to initially understand the effect of entB on CRKP colony morphology and virulence phenotype, the colony morphology of CRKP-27, Δ entB, and C-Δ entB strains were observed and string test were tested. Draw the growth curve of the strains and determine the effect of entB on the growth of the CRKP strains. The siderophores production ability of the strains were detected quantitatively using chrome azurol S (CAS) detection solution. Mice model of inflammation was established to observe the survival rate of mice and intuitively understand the effect of entB on CRKP virulence. Results:The PCR results showed that the Δ entB strain and C-Δ entB stranin were constructed successfully. The entB has no significant effect on the colony morphology, capsule and virulence phenotype of CRKP. The growth rate of Δ entB was significantly faster than that of CRKP-27( P=0.008) and C-Δ entB ( P=0.001), which showed that entB weakened the growth ability of CRKP. Compared with CRKP-27( P=0.001) and C-Δ entB( P=0.001), the siderophore production of Δ entB was significantly decreased by 11.739 3% and 11.964 2%, indicating that entB gene increased the capacity of CRKP to produce siderophpres. In animal experiments, compared with CRKP-27( P=0.023) and C-Δ entB( P=0.024), the survival rate of mice in the Δ entB group was significantly increased, indicating that the entB increased the virulence of the CRKP. Conclusion:The siderophore virulence gene entB significantly weakened the growth ability of the strain, but clearly enhanced the siderophore production capacity and virulence of CRKP.

Objective: To observe theclinical effect of tuina reduction manipulation on leg length discrepancy and lumbosacral pain due to sacroiliac joint subluxation.
Methods: A total of 60eligible cases were randomly allocated into an observation group and a control group, 30 in each group. Cases in the observation group were treated with conventional tuina plus reduction manipulation of sacroiliac joint subluxation; whereas cases in the control group were treated with conventional tuina plus acupuncture. The clinical effects were observed after 10 times of treatment. In addition, the relapse rates were observed 2 months after treatment.
Results: The total effective rate in the observation group was 80.0%, versus 50.0% in the control group, showing a statistically significant difference (P<0.05). The relapse rate of lumbosacral pain in the observation group was 12.5%, versus 66.7% in the control group, showing a statistically significant difference (P<0.01). The relapse rate of leg length discrepancy in the observation group was 16.7%, versus 80.0% in the control group, showing a statistically significant difference (P<0.01).
Conclusion: Tuina reduction manipulation can obtain substantial therapeutic effect for leg length discrepancy and lumbosacral pain due to sacroiliac joint subluxation, coupled with a low relapse rate.