Efficacy of umbilical cord mesenchymal stem cells replacing donor bone marrow cells in haploidentical hematopoietic stem cell transplantation / 中国组织工程研究

BACKGROUND:HLA haploid allogeneic hematopoietic stem cell transplantation provides a chance of survival for patients with high-risk hematologic malignancies.In recent years,the research on the transplantation mode and graft selection of haploidentical transplantation is still ongoing.At present,the mixed transplantation model of non-extracorporeal T-cell removal bone marrow and peripheral blood stem cells established by the Hematology Research Center of Peking University is gradually becoming popular in China,but this model requires the collection of donor bone marrow fluid,which increases the pain and risk of the donor. OBJECTIVE:To explore the curative effect of infusion of umbilical cord mesenchymal stem cells replacing donor bone marrow cells in haploidentical peripheral blood hematopoietic stem cell transplantation for malignant hematological diseases. METHODS:Fifty hematological malignancies patients who underwent haploidentical hematopoietic stem cell transplantation from January 2019 to May 2022 were selected and randomly assigned to two study groups at a ratio of 2:3.Among them,19 patients received umbilical cord mesenchymal stem cell combined with peripheral blood stem cell transplantation,and 31 patients were treated with bone marrow cells combined with peripheral blood stem cells.The study was approved by the Ethics Committee of Henan Provincial People's Hospital.The recipients of umbilical cord mesenchymal stem cells were first transfused with third-party umbilical cord mesenchymal stem cells(1×106/kg)on the day of transplantation,followed by peripheral blood hematopoietic stem cells 6 hours later.In the bone marrow group,donor bone marrow cells were transfused +1 day after transplantation and peripheral blood stem cells were transfused +2 days after transplantation.After transplantation,rabbit anti-human thymocyte immunoglobulin,cyclosporine A,mycophenolate mofetil,and a short-course methotrexate were used for graft-versus-host disease prophylaxis for all recipients. RESULTS AND CONCLUSION:No adverse events occurred during the reinfusion of umbilical cord mesenchymal stem cells.There were no significant differences between the mesenchymal stem cell group and the bone marrow group in the engraftment rate[100%(19/19)vs.96.8%(30/31),P>0.05],median duration for neutrophil engraftment(14 days vs.15 days,P>0.05)and median duration for platelet engraftment(20 days vs.19 days,P>0.05).The incidence of grade Ⅱ-Ⅳ acute graft-versus-host disease in the mesenchymal stem cell group was significantly lower than in the bone marrow group[21.1%(4/19)vs.58.1%(18/31),P = 0.01].There were no significant differences between the two groups in the incidence of chronic graft-versus-host disease[21.1%(4/19)vs.25.8%(8/31),P>0.05],the relapse rates[15.8%(3/19)vs.16.1%(5/31),P>0.05]and the incidence of early cytomegalovirus viremia[42.1%(8/19)vs.35.5%(11/31),P>0.05],and the 2-year overall survival rate[68.4%(10/19)vs.70.9%(16/31),P>0.05].It is indicated that umbilical cord mesenchymal stem cells replace donor bone marrow cells in haploidentical peripheral blood stem cell transplantation for malignant hematological diseases,which reduced the incidence of acute graft-versus-host disease after transplantation,did not increase the incidence of chronic graft-versus-host disease,recurrence rate and early cytomegalovirus viremia,and reduced the pain and risk of donor pulp extraction.

Correlation between SLAMF6 expression and perforin and granzyme B on circulating CD8 +T cells and the clinical significance in patients with severe aplastic anemia / 中华检验医学杂志

Objective:To explore the correlation between the expression of signaling lymphocyte activation molecule family 6 (SLAMF6) on peripheral blood CD8 +T cells and perforin and granzyme B and the clinical significance in patients with newly diagnosed severe aplastic anemia(SAA). Methods:The indicators of blood routine and bone marrow and peripheral blood samples of 32 newly diagnosed SAA patients admitted to Henan Provincial People′s Hospital from January 2016 to June 2019 were collected for retrospective analysis. Flow cytometry was used to detect the expression of SLAMF6, perforin and granzyme B on samples CD8 +T cell before therapy and 6 months after therapy (11 cases received transplantation, 21 cases received immunosuppressive therapy [IST]). Spearman correlation analysis was performed to determine the association between clinical indicators and laboratory test results. The expression of SLAMF6, perforin and granzyme B was also detected in 10 healthy people (normal group) and 13 myelodysplastic syndromes/paroxysmal nocturnal hemoglobinuria (MDS/PNH) patients (MDS/PNH group). Results:(1) At diagnosis: the expression of SLAMF6 was significantly lower in the SAA group than that in the normal group and the MDS/PNH group ([56.40±6.37]% vs [84.34±5.81]% and [82.24±4.98]% (both P<0.001]). The expression of perforin was significantly higher in the SAA group (32.73±8.46) than that in the normal control group (23.75%±5.10%), and the MDS/PNH group (26.12%±5.53%) (both P<0.05). The expression of granzyme B was also significantly higher in the SAA group (36.23%±7.94%) than that in the normal control group (21.67%±5.05%) and the MDS/PNH group (21.79%±5.10%) (both P<0.001). The expression of SLAMF6 was positively correlated with the hemoglobin ( r=0.804), and reticulocyte absolute values ( r=0.656) in peripheral blood, percentage of granulocytes ( r=0.643) and erythrocytes ( r=0.622) in bone marrow of SAA patients (all P<0.05). Expression of SLAMF6 was negatively correlated with perforin ( r=-0.792) and granzyme B ( r=-0.908) on CD8 +T cells in patients with SAA (both P<0.001). (2) After treatment: the expression of SLAMF6 in peripheral blood CD8 +T cells of 30 surviving patients was higher than pre-treatment ([79.19±12.69]% vs [56.40±6.37]%, P<0.001). The expressions of perforin and granzyme B were lower than pre-treatment level (both P<0.05). The expression of SLAMF6 on CD8 +T cells in 11 transplanted patients was higher than before transplantation ([86.54±3.75]% vs [56.40±7.35]%, P<0.001). The expressions of perforin and granzyme B were lower than before transplantation (both P<0.05). The expression of SLAMF6 on CD8 +T cells in 12 IST-respond patients was higher than that before treatment, while the perforin and granzyme B levels were lower than pre-treatment (all P<0.05). The post-treatment expressions of SLAMF6, perforin and granzyme B were similar as before treatment levels in 7 IST-unrespond patients (all P>0.05). Conclusion:SLAMF6 is significantly down-regulated on CD8 +T cells in newly diagnosed SAA, negatively correlated with the effective factors of CD8 +T cells, which might participate in the immune regulatory of CD8 +T cells as a negative regulatory factor in patients with SAA. The SLAMF6 is significantly up-regulated after hematopoietic recovery, while there is no significant change in treatment-unrespond patients, which could thus serve as an useful diagnostic and therapeutic index of patients with SAA.

Expression of CD45 in newly diagnosed multiple myeloma and the relationship with prognosis / 中华血液学杂志

Objective@#To explore the expression of CD45 in newly diagnosed multiple myeloma (MM) and its relationship with clinical efficacy and prognosis.@*Methods@#This study retrospectively analyzed expression and distribution of CD45 in 130 cases of newly diagnosed MM, comparing clinical efficacy and prognosis in CD45+/CD45- groups.@*Results@#①The CD45+ group was 33 cases (25.38%) , and CD45- group was 97 cases (74.62%) . ②The objective remission rate (ORR) of CD45+ and CD45-group was 33.33% and 64.95%, respectively. The difference was statistically significant (P=0.002) . For patients in Bortezomib regimen, the ORR of CD45+ and CD45- group was 35.71% and 66.25%, respectively. The difference was statistically significant (P=0.005) . ③The median progress free survival (PFS) of CD45+ group and CD45- group was 29.8 (95%CI 10.0-59.0) months vs 34.5 (95%CI 6.0-69.0) months (χ2=14.59, P<0.001) and the median overall survival (OS) was 32.5 (95%CI 10.0-68.0) months vs 37.6 (95%CI 6.0-78.0) months (χ2=11.42, P=0.001) , respectively. Among the patients in bortezomib regimen, The median PFS and median OS of CD45 + group and CD45- group were 30.3 (95%CI 10.0-59.0) months vs 36.3 (95%CI 6.0-69.0) months (χ2=14.75, P=0.001) and 34.0 (95%CI 10.0-68.0) months vs 39.5 (95%CI 6.0-78.0) months (χ2=10.62, P=0.001) . ④Cox risk regression model analysis showed that serum creatinine≥176.8 μmol/L (HR=5.078, 95%CI 1.744-14.723, P=0.001) , CD45 positive (HR=14.504, 95%CI 0.168-0.42, P=0.001) , LDH≥220 IU/L (HR=1.308, 95%CI 1.16-2.417, P=0.015) were independent risk prognostic factors.@*Conclusion@#CD45 expression is a risk prognostic factor of MM patients. Bortezomib did not improve the poor prognosis of CD45+ MM patients.

Kinetics investigation of the hydrogen abstraction reaction between CH3SS and CN radicals.

The reaction mechanisms and rates for the H abstraction reactions between CH3SS and CN radicals in the gas phase were investigated with density functional theory (DFT) methods. The geometries, harmonic vibrational frequencies, and energies of all stationary points were obtained at B3PW91/6-311G(d,p) level of theory. Relationships between the reactants, intermediates, transition states and products were confirmed, with the frequency and the intrinsic reaction coordinate (IRC) analysis at the same theoretical level. High accurate energy information was provided by the G3(MP2) method combined with the standard statistical thermodynamics. Gibbs free energies at 298.15 K for all of the reaction steps were reported, and were used to describe the profile diagrams of the potential energy surface. The rate constants were evaluated with both the classical transition state theory and the canonical variational transition state theory, in which the small-curvature tunneling correction was included. A total number of 9 intermediates (IMs) and 17 transition states (TSs) were obtained. It is shown that IM1 is the most stable intermediate by the largest energy release, and the channel of CH3SS + CN → IM3 → TS10 → P1(CH2SS + HCN) is the dominant reaction with the lowest energy barrier of 144.7 kJ mol(-1). The fitted Arrhenius expressions of the calculated CVT/SCT rate constants for the rate-determining step of the favorable channel is k =7.73 × 10(6) T (1.40)exp(-14,423.8/T) s(-1) in the temperature range of 200-2000 K. The apparent activation energy E a(app.) for the main channel is -102.5 kJ mol(-1), which is comparable with the G3(MP2) energy barrier of -91.8 kJ mol(-1) of TS10 (relative to the reactants).

The improved application of artificial dermaiin repairing depth of burn wounds / 中国综合临床

ObjectiveTo explore the therapeutic efficacy of artificial dermal in repairing depth of burn wounds.MethodsTwenty-two cases of patients with depth bum who were admitted to our hospital during August of 2008 to August of 2010 were enrolled in this trial for retrospective study.The style of management was evaluated in these patients.Burn wound depth and severity was assessed immediately after patients admission,wound treatment was performed after patients with stable vital signs and wound edema peak period vanished.Artificial dermal was grafted onto the wound tissue after the primary debridement. Then,transplantation of artificial dermal was performed,with moist dressingonto it.After two weeks,the razor thick autoskin was grafted onto the surface of the artificial dermal after the removing of its silicon membrane during the secondary operation.ResultsThe wound healed completely with the survival of skin grafting and satisfactory appearance in all 22 patients. Furthermore,the scar in the donor site exhibited unconspicuous. Conclusion It's an optimal choice to repair deep burn with artificial dermal and thick autoskin.

Comparison between clinical results of peripheral blood stem cell transplantation from HLA-matched sibling donors mobilized with different regimens / 中华器官移植杂志

Objective To retrospectively review and compare the clinical results of allogeneic peripheral blood stem cell transplantation (allo-PBSCT) from HLA- matched sibling donors mobilized with different regimens. Methods Seventy-one patients with hematological malignant diseases received allo-PBSCT from HLA-matched sibling donors in our department. Among them, 24 received allografts mobilized with G-CSF (group G), and the remaining (47 cases) were mobilized with G-CSF and GM-CSF (group G+ M). CD34+ subsets and T cell subsets in the allografts were analyzed, and the time of hematopoietic reconstitution and the incidence of graft versus host diseases (GVHD) were compared. The adverse effects on the donors after mobilization were also observed. Results The enough targeted CD34+ cells in all donors were harvested by 1-3 aphereses. Ninety-six h after mobilization, WBC counts of the donors were significantly higher in group G than in group G + M [(49. 6± 19. 5) 109/L vs (25.4 ± 10. 4) 109/L, P＜0. 05]. Analysis of the CD34+ subsets showed that the percentage of cells with the CD34+/CD38- phenotype was significantly higher in group G + M than in group G [(37. 7 ± 5. 7) % vs (31.4 ± 4. 5) %, P＜0. 05]. There was no significant difference in T cells and subsets of grafts. There was no significant difference in the number of total CD34+ cells and CD34+ CD38- cells, and infusion of T cells between two groups. The days required for the recovery of neutrophils and platelets was inversely correlated with the infused CD34+ and CD34+ /CD38- cell number. There was no significant difference in incidence of acute and chronic GVHD between two recipient groups. Seventeen cases and 10 eases among 71 eases died of relapses of primarydiseases, and complications of transplantation such as severe GVHD and infections respectively.Fourteen cases in group G (58.3 %) and 31 cases in group G+ M (66.0 %) survived. The most common adverse events in the donors were bone pain and fever, which mostly occurred 36 h after mobilization and could be relieved by non-steroidal anti-inflammatory drugs. Conclusion Two mobilization regimens showed equivalent clinical results. But the combined regimen of G-CSF and GM-CSF demonstrated a significantly greater mobilization of cells with the CD34+/CD38- phenotype.Meanwhile in allogeneic PBSCT, a greater number of total CD34+ cells and CD34+ CD38- cells infused may be associated with faster hematopoietic reconstitution of recipients.

Assessment of static and dynamic balance in hemiparetic stroke patients / 中国康复理论与实践

@#ObjectiveTo discusse the relationship between static balance in laboratory approaches and dynamic balance in clinical assessment and identify the value of static and dynamic balance at functional outcome in hemiparetic stroke patients. MethodsNineteen stroke subjects were assessed in this study. The static balance was measured by postural sway test, the dynamic balance was measured by Berg balance scale(BBS)and Time up to go test(TUGT),the outcome was measured by FIM and 10m maximum walking speed(MWS). The level of association between the parameters of postural sway test and clinical variables were examined with Pearson's correlation coefficients. ResultsThe parameters of postural sway test was significantly negative related to BBS(r=-0.705--0.475,P<0.05);The parameters of postural sway close-eye test was significantly positive related to TUGT(r=0.508-0.583,P<0.05);The parameters of postural sway test was no related to FIM and MWS (r=-0.048--0.296;r=-0.404--0.01,P>0.05);BBS was significantly positive related to FIM and MWS(r=0.752;r=0.700,P<0.001). TUGT was significantly negative related to FIM and MWS(r=-0.600,P<0.01;r=-0.817,P<0.001).ConclusionClinical and laboratory balance assessments are related and that dynamic rather than static balance measures are valid indicators of functional outcome performance in hemiparetic stroke patients.

Value of walking speed at the evaluation of motor function recovery in hemiparetic patients after stroke / 中国康复理论与实践

@#ObjectiveTo identify the value of walking speed at the evaluation of motor function recovery in hemiparetic patients after stroke.MethodsFifty-five cases were assessed in the study. The walking speed, motor function, strengths of the paretic lower limb,balance and ambulation of ADL function were evaluated with the 10m Maximum Walking Speed Test,Fugl-Meyer Assessment, Motricity Index, Berg Balance Scale and ambulation item of Functional Independence Measure. The level of association between walking speed and the clinical variables were examined with Pearson's correlation coefficients.ResultsIn the slow group, the gait-related clinical scores were related to walking speed (Pearson's r=0.581-0.770,P<0.05),while that in the moderate group and fast group were not (Pearson's r=0.016-0.380,P>0.05).The correlation between the gait-related clinical scores and walking speed was weakened when walking speed above 44.8m/min(or 41.3% of normal values).ConclusionWalking speed plays an independent role as an indicator of improved performance.

Selective depletion of the allo-antigen specific T cells by Fas/FasL pathway by cytokine IFN-gamma and IL-2 / 华中科技大学学报（医学）（英德文版）

To investigate the value of apoptosis of the allo-antigen specific T cells induced by Fas/FasL pathway in preventing graft-versus-host disease (GVHD), the CD34+ cells transfected with FasL or not, used as stimulus cells, were mixed with allo-antigen specific T lymphocytes in presence or absence of IFN-gamma and IL-2. After 5 days, apoptosis of T cells was detected by TdT nick end mediated dUTP labeling (TUNEL) and flow cytometry (FCM). The affects of these two cytokines on CD34+ cells in the graft were also compared. The ratio of apoptosis of T cells was 12.1+/-1.5% when CD34+ cells transfected with FasL was used as stimulus cells, much higher than that of CD34+ cells non-transfected (3.2+/-1.1%, P<0.01). And in presence of IFN-gamma or IL-2, the ratio reached 20.1+/-2.3%, 17.6+/-1.3% respectively (P<0.01). However, IFN-gamma up-regulated Fas expression of CD34+ cells and increased the sensibility of CD34+ cells to soluble FasL (sFasL); IL-2 showed no such effect. It is possible to induce apoptosis of the allo-antigen specific T cells of grafts activated by allo-antigen by exogenous Fas ligand expressed on recipient cells and this might provide a new approach for preventing GVHD and IL-2 may be more suitable for clinical application.

Apoptosis of leukemia cells induced by CD34+ cells transferred exogenous Fas ligand / 华中科技大学学报（医学）（英德文版）

To assess the value of CD34+ cells transferred exogenous Fas ligand (FasL) in inducing apoptosis of human leukemic cells, the CD34+ cells transfected with FasL or without, pretreated with mitomycin C, was mixed with leukemic cell line U937 cells in presence or absence of daunorubicin (DNR) or cytosine arabinoside (Ara-C). After 18 h, apoptosis of cells was detected by FCM and TUNEL. Induced for 18 h by CD34+ cells transfected with FasL or without, the ratio of apoptosis of U937 cells was (5.0 +/- 1.3)%, (10.8 +/- 0.6)% (P < 0.01), respectively. Induced by FasL+ CD34+ + DNR, FasL+ CD34+ + Ara-C, the ratio was (13.4 +/- 1.0)% (P < 0.05), (17.9 +/- 1.3)% (P < 0.01), respectively. The result demonstrated that CD34+ cells transfected with exogenous FasL could induce apoptosis of human leukemic cells and showed a cytotoxic synergistic effect when used in combination with chemotherapeutic drugs, suggesting that it was possible to develop a new method in treatment of leukemia.