Effects of donor cells on in vitro development of cloned bovine embryos

The donor cells from different individuals and with different foreign genes introduced were investigated to determine their effects on the efficiency of somatic cell nuclear transfer （SCNT）. The bovine ear fibroblast from different individuals was isolated, cultured, and then transfected with foreign genes to establish the stable cell lines, which were used as donor cells for nuclear transfer. The oocytes were obtained through ovum pick up operation. After in vitro maturation, the M II phase oocytes were selected as receptors for nuclear transfer. The reconstructed embryos were cultured in vitro and observed at 2 h, 48 h, and 7 days after transfer to assess the rate of fusion using cleaved and blastocyst as the parameters of SCNT efficiency. The donor cells from different individuals （04036, 06081, 06088, and 06129） had no obvious effect on the fusion and cleaved rate, whereas there was significant difference in the blastocyst rate （P〈0.05）, and the rate was 62.3%, 37.0%, 35.1%, and 15.6%, respectively. There was no significant difference among the rate of fusion, cleaved and blastocyst in donor cells with different foreign genes （P〉0.05）. It was concluded that the genetic background of the donor cells could affect the efficiency of SCNT, while the introduction of foreign genes into the donor cells had no obvious effect on the efficiency. This study provides useful information for the SCNT and would benefit in promoting the efficiency.

Sickle Cell Disease in the Zinder Region in 2023: Prevalence and Sociodemographic Factors

Introduction: In view of the number of sickle cell patients and due to a low production of descriptive studies, we decided to determine the prevalence of genes S and C of the disease in the Zinder region. The objective was to contribute to improving the management of sickle cell disease in Zinder. Methodology: This was a systematic screening by the “Sickle Scan” test of any blood donor admitted to the Zinder Regional Blood Transfusion Center during the 6-month study period, from January to June 2023. The Sickle Scan is a qualitative lateral flow chromatography immunoassay using whole blood samples that aid in the rapid diagnosis of sickle cell disease. Results: The study was carried out on 613 samples during the period concerned. The frequency of sickle cell genes was 26.9% (n = 165) in all samples collected, with 23.1% (n = 142) and 3.8% (n = 23) for the S gene and the C gene, respectively. The 18 - 30 age group was the most represented with 64.4% (n = 395) cases. The median age of blood donors was 26 years ± 10 years (min = 18 years/max = 60 years). The sex ratio was 2.5. Donors of Nigerien nationality accounted for 84.1% (n = 516). There is a predominance of blood donors with an average monthly income between 34,000 and 70,000 CFA francs in 44.3% (n = 272), lived in permanent housing with drinking water supply. Sickle cell trait (SMA) was found in 22.5% (n = 138). Conclusion: The analysis of these results highlights a high frequency of the S gene for sickle cell disease. The population with an average monthly income is the most affected, with a male predominance.

Safety of in vitro amplified HLA-haploidentical donor immune cell infusions for childhood malignancies

In vitro amplified human leukocyte antigen(HLA)-haploidentical donor immune cell infusion(HDICI)is not commonly used in children.Therefore,our study sought to evaluate its safety for treating childhood malignancies.Between September 2011 and September 2012,12 patients with childhood malignancies underwent HDICI in Sun Yat-sen University Cancer Center.The median patient age was 5.1 years(range,1.7-8.4 years).Of the 12 patients,9 had high-risk neuroblastoma(NB)[7 showed complete response(CR),1 showed partial response(PR),and 1 had progressive disease(PD)after multi-modal therapies],and 3 had Epstein-Barr virus(EBV)-positive lymphoproliferative disease(EBV-LPD).The 12 patients underwent a total of 92 HDICIs at a mean dose of 1.6×108immune cells/kg body weight:71 infusions with natural killer(NK)cells,8 with cytokine-induced killer(CIK)cells,and 13 with cascade primed immune cells(CAPRIs);83 infusions with immune cells from the mothers,whereas 9 with cells from the fathers.Twenty cases(21.7%)of fever,including 6 cases(6.5%)accompanied with chills and 1(1.1%)with febrile convulsion,occurred during infusions and were alleviated after symptomatic treatments.Five cases(5.4%)of mild emotion changes were reported.No other adverse events occurred during and after the completion of HDIDIs.Neither acute nor chronic graft versus host disease(GVHD)was observed following HDICIs.After a median of 5.0 months(range,1.0-11.5 months)of follow-up,the 2 NB patients with PR and PD developed PD during HDICIs.Of the other 7 NB patients in CR,2 relapsed in the sixth month of HDICIs,and 5 maintained CR with disease-free survival(DFS)ranging from 4.5 to 11.5 months(median,7.2months).One EBV-LPD patient achieved PR,whereas 2 had stable disease(SD).Our results show that HDICI is a safe immunotherapy for childhood malignancies,thus warranting further studies.

Chronic ocular GVHD:limbal and conjunctival stem cell allografts from the same hematopoietic stem cell donor

Dear Editor,I am Dr.Elias F Jarade from the Beirut Eye and ENT Specialist Hospital, Beirut, Lebanon. I write to describe a novel surgical technique in the management of chronic ocular graft-versus-host disease（GVHD）.

Donor liver natural killer cells alleviate liver allograft acute rejection in rats

BACKGROUND:Liver enriched natural killer (NK) cells are of high immune activity.However,the function of donor liver NK cells in allogeneic liver transplantation (LTx) remains unclear.METHODS:Ten Gy of whole body gamma-irradiation (WBI) from a 60 Co source at 0.6 Gy/min was used for depleting donorderived leukocytes,and transfusion of purified liver NK cells isolated from the same type rat as donor (donor type liver NK cells,dtl NKs) through portal vein was performed immediately after grafting the irradiated liver.Post-transplant survival observation on recipients and histopathological detection of liver grafts were adoptive to evaluate the biological impact of donor liver NK cells on recipients’ survival in rat LTx.RESULTS:Transfusion of dtl NKs did not shorten the survival time among the recipients of spontaneous tolerance model (BN to LEW rat) after rat LTx,but prolonged the liver graft survival among the recipients depleted of donor-derived leukocytes in the acute rejection model (LEW to BN rat).Compared to the recipients in the groups which received the graft depleted of donor-derived leukocytes,better survival and less damage in the allografts were also found among the recipients in the two different strain combinations of liver allograft due to transfusion of dtl NKs.CONCLUSIONS:Donor liver NK cells alone do not exacerbate liver allograft acute rejection.Conversely,they can alleviate it,and improve the recipients’ survival.

Novel donor-acceptor-donor structured small molecular hole transporting materials for planar perovskite solar cells

Novel donor-acceptor-donor structured small molecular hole transporting materials are developed through a facile route by crosslinking dithienopyrrolobenzothiadiazole and phenothiazine or triarylamine-based donor units. The strong push/pull electron capability of dithienopyrrolobenzothiadiazole/ phenothiazine and large π-conjugated dithienopyrrolobenzothiadiazole facilitate hole mobility and high conductivity. The devices using the dithienopyrrolobenzothiadiazole/phenothiazine-based hole trans-porting material achieved a power conversion efficiency of 14.2% under 1 sun illumination and improved stability under 20% relative humidity at room temperature without encapsulation. The present finding highlights the potential of dithienopyrrolobenzothiadiazole-based donor-acceptor-donor small molecular hole transporting materials for perovskite solar cells.

Analysis of CD8^+CD28^-T-suppressor cells in living donor liver transplant recipients

BACKGROUND:Human CD8 + CD28 - T-suppressor(Ts) cells have been considered to indicate a reduced need for immunosuppression in pediatric liver-intestine transplant recipients and recipients of deceased heart-kidney transplants.However,in adult-to-adult living donor liver transplantation(A-A LDLT)little information is available and the clinical significance is still unknown. METHODS:Flow cytometry was used to detect the population of CD8+CD28 -Ts cells present in peripheral blood in A-A LDLT recipients(n=31),patients with end- stage liver disease(n=24)and healthy controls(n=19). Meanwhile,we tested the graft function and trough levels of immunosuppression in recipients.The clinical and follow- up data of 31 transplant recipients were analyzed. RESULTS:Compared with diseased controls(P=0.007) and healthy individuals(P=0.000),a notable expansion of CD8 + CD28 - Ts cells was found in recipients of A-A LDLT.This was associated with graft function,levels of immunosuppression and rejection episodes. CONCLUSIONS:To monitor the CD8 + CD28 - Ts cells levels is important to evaluate the immune state of recipients. Meanwhile,it is also important to promote expansion of CD8+CD28 -Ts cells in recipients of A-A LDLT,not only to sustain good graft function and decrease the dosage of immunosuppressants,but also to reduce the occurrence of rejection.

High-performance asymmetric small molecular donor materials based on indenothiophene for solution-processed organic solar cells

Two novel asymmetric organic small molecules of IT(2FBT-T3Cz)_2and IT(2FBT-TT3Cz)_2with an indenothiophene(IT)central donor core,fluorinated benzothiadiazole(2FBT)as acceptor and 3-carbazole(Cz)unit as terminal group were designed and synthesized as the donor materials in organic solar cells(OSCs).The thermal,optical absorption,electrochemical property,hole–electron mobility,film morphology were thoroughly studied.Using PC_(71)BM as an electron acceptor,without any additive and thermal annealing(TA)treatment,the IT(2FBT-T3Cz)_2-based cells showed a promising power conversion efficiency(PCE)of5.81%and the IT(2FBT-TT3Cz)_2-based cells exhibited a PCE of 4.39%.Our results demonstrate that the IT-based asymmetric small molecules can be developed as a promising class of donor materials for highperformance OSCs.

Preoperative application of combination of portal venous injection of donor spleen cells and intraperitoneal injection of rapamycin prolongs the survival of cardiac allografts in mice

Objective:To investigate the effects of preoperative portal venous injection of donor spleen cells(PVIDSC) and intraperitoneal injection of rapamycin in the acute rejection of cardiac allograft in mice and the underlying mechanisms. Methods:Homogenous female B6 mice and BALB/c mice were used as recipients and donors of heart transplantation. These mice were randomly divided into different groups and received PVIDSC alone,rapamycin alone,or PVIDSC and rapamycin combined therapy. In addition,the underlying mechanism was studied by measuring a number of cytokines. Results:Preoperative combination of PVIDSC and intraperitoneal injection of rapamycin significantly prolonged the survival of heterotopic cardiac allograft in mice,but had no effects on the survival time of cardiac allografts in mice pre-sensitized by skin grafting. Preoperative combination of PVIDSC and intraperitoneal injection of rapamycin increased the expression of IL-10 and Foxp3 and reduced the expression of INF-γ. Short-term preoperative administration of rapamycin promotes the expression of CD4^+CD25^+Foxp3^+ regulator T cells. However,preoperative using alone of rapamycin,or combination of PVIDSC and rapamycin had no effects on the inhibition of proliferation of memory T cells. Conclusions:Preoperative application of combination of PVIDSC and rapamycin significantly prolonged the survival time of cardiac allografts in mice but not in mice pre-sensitized by skin grafting. This may be explained by the fact that combination of PVIDSC and rapamycin inhibited the cellular immune response and induced the expression of IL-10 from Tr1 cells and CD4^+CD25^+FoxP3^+ regulatory T cells.

Donor-Derived CD19-Targeted T Cell Infusion Eliminates B Cell Acute Lymphoblastic Leukemia Minimal Residual Disease with No Response to Donor Lymphocytes after Allogeneic Hematopoietic Stem Cell Transplantation

Leukemia relapse is still the leading cause of treatment failure after allogeneic hematopoietic stem cell transplantation (allo-HSCT) for B cell acute lymphoblastic leukemia (B-ALL). Relapsed patients with BALL after allo-HSCT have a very short median survival. Minimal residual disease (MRD) is predictive of forthcoming hematological relapse after hematopoietic stem cell transplantation (HSCT);furthermore, eliminating MRD effectively prevents relapse. Donor lymphoblastic infusion (DLI) is the main established approach to treat B-ALL with MRD after allo-HSCT. However, about one-third of patients with MRD are non-responsive to DLI and their prognosis worsens. Although donor-derived cluster of differentiation (CD)19-directed chimeric antigen receptor-modified (CAR) T cells (CART19s) can potentially cure leukemia, the efficiency and safety of infusions with these cells have not yet been investigated in patients with MRD after HSCT. Between September 2014 and February 2018, six patients each received one or more infusions of CART19s from HSCT donors. Five (83.33%) achieved MRD-negative remission, and one case was not responsive to the administration of CAR T cells. Three of the six patients are currently alive without leukemia. No patient developed acute graft-versus-host disease (aGVHD), and no patient died of cytokine release syndrome. Donor-derived CAR T cell infusions seem to be an effective and safe intervention for patients with MRD in B-ALL after allo-HSCT and for those who were not responsive to DLI.

Application of donor dendric cell mediated recipieut lymphocyte reaction after related kidney transplantation in individualized immunosuppressive therapy

Objective To explore the feasibility of mediating recipient lymphocyte reaction with donor dendritic cells ( Dcs) in renal allograft recipients to guide individualized immunosuppressive therapy. Methods From Jan. 2008 to Jan. 2010,30 recipients received living related kidney transplantation were successively and divided into

Unsymmetrical donor—acceptor—donor—acceptor type indoline based organic semiconductors with benzothiadiazole cores for solution-processed bulk heterojunction solar cells

Bulk heterojunction(BHJ) solar cells based on small molecules have attracted potential attention due to their promise of conveniently defined structures, high absorption coefficients, solution process-ability and easy fabrication. Three D—A—D—A type organic semiconductors(WS-31,WS-32 and WS-52) are synthesized, based on the indoline donor and benzotriazole auxiliary acceptor core, along with either bare thiophene or rigid cyclopentadithiophene as π bridge, rhodanine or carbonocyanidate as end-group. Their HOMO orbitals are delocalized throughout the whole molecules. Whereas the LUMOs are mainly localized on the acceptor part of structure, which reach up to benzothiadiazole, but no distribution on indoline donor. The first excitations for WS-31 and WS-32 are mainly originated by electron transition from HOMO to LUMO level, while for WS-52, partly related to transition between HOMO and LUMO+1 level. The small organic molecules are applied as donor components in bulk heterojunction(BHJ) organic solar cells, using PC_(61)BM as acceptor material to check their photovoltaic performances. The BHJ solar cells based on blended layer of WS-31:PC_(61)BM and WS-32:PC_(61)BM processed with chloroform show overall photoelectric conversion efficiency(PCE) of 0.56% and 1.02%, respectively. WS-32 based BHJ solar cells show a higher current density originated by its relatively larger driving force of photo-induced carrier in photo-active layer to LUMO of PC_(61)BM.

单采血小板献血者发生红细胞溢出的护理1例

目的探讨单采血小板献血者发生红细胞溢出的原因分析和护理思路,避免献血不良反应或单采血小板报废,提升献血服务水平。方法依据文献证据制定红细胞溢出的护理流程图,附单采血小板献血者发生红细胞溢出的护理过程1例,并进行相应的文献复习。结果通过红细胞溢出的护理流程图施行护理干预后,献血者感受良好未发生献血不良反应,未再出现红细胞溢出,血小板成功采集。通过复习相关文献,系统性总结红细胞溢出的发生率、原理、原因、处理方法、预防献血不良反应及心理护理方法等。结论单采血小板献血者红细胞溢出偶有发生,红细胞溢出的护理流程图可帮助血站工作人员快速鉴别出原因并正确处理,避免处理不当造成不良反应或单采血小板报废,以获得良好结局。

Brentuximab Vedotin Monotherapy and Combined with Low Dose Donor Lymphocyte Infusion to Control Minimal Residual Disease and Sustain Clinical Remission in a Child with Relapsed Anaplastic Large Cell Lymphoma

Minimal residual disease (MRD) appears to have a strong negative predictive value for disease recurrence in children with anaplastic large cell lymphoma (ALCL). Brentuximab vedotin (BV) can be a therapeutic option for MRD-positive patients to achieve molecular remission and to decrease risk of subsequent relapse. We here report a 4-year-old child with ALCL progression during relapse treatment who received BV as a bridging therapy before haploidentical hematopoietic stem-cell transplantation, and as a maintenance therapy post-transplant alone or combined with simultaneous low dose donor-lymphocyte infusions. MRD monitoring showed a complete molecular response and reflected both BV efficiency and graft-versus-lymphoma effect.

T-cell allorecognition of donor glutathione S-transferase T1 in plasma cell-rich rejection

AIM To investigate the role of glutathione S-transferase T1 donor-specific T lymphocytes in plasma cell-rich rejection of liver allografts.METHODS The study group included 22 liver transplant patients. Among them, 18 patients were mismatched for the glutathione S-transferase T1(GSTT1) alleles(don+/rec-), and 4 were matched(don+/rec+). Seven of the mismatched patients produced anti-GSTT1 antibodies and developed plasma cell-rich rejection(former de novo immune hepatitis). For the detection of specific Tlymphocytes, peripheral blood mononuclear cells were collected and stored in liquid nitrogen. The memory T cell response was studied by adding to the cell cultures to a mix of 39 custom-made, 15-mer overlapping peptides, which covered the entire GSTT1 amino acid sequence. The specific cellular response to peptides was analyzed by flow cytometry using the markers CD8, CD4, IL-4 and IFNγ.RESULTS Activation of CD8^+ T cells with different peptides was observed exclusively in the group of patients with plasma-cell rich rejection(3 out of 7), with production of IL-4 and/or IFNγ at a rate of 1%-4.92% depending on the peptides. The CD4^+ response was most common and not exclusive for patients with the disease, where 5 out of 7 showed percentages of activated cells from 1.24% to 31.34%. Additionally, two patients without the disease but with the mismatch had cells that became stimulated with some peptides(1.45%-5.18%). Highly unexpected was the finding of a double positive CD4^+CD8^(low) T cell population that showed the highest degree of activation with some of the peptides in 7 patients with the mismatch, in 4 patients with plasma cell-rich rejection and in 3 patients without the disease. Unfortunately, CD4^+CD8^(low) cells represent 1% of the total number of lymphocytes, and stimulation could not be analyzed in 9 patients due to the low number of gated cells. Cells from the 4 patients included as controls did not show activation with any of the peptides. CONCLUSION Patients with GSTT1 mismatch can develop a specific T-cell response, but the potential role of this response in the pathogenesis of plasma cell-rich rejection is unknown.

双重膜滤过式血浆置换联合脱敏治疗单倍体相合造血干细胞移植供者特异性抗体的临床研究

目的探讨分析双重膜滤过式血浆置换(double filtration plasmapheresis,DFPP)联合脱敏治疗单倍体相合造血干细胞移植供者特异性抗体(donor specific antibody,DSA)的疗效。方法采用DFPP联合丙种免疫球蛋白(intravenous immunoglobulin,IVIG)、利妥昔单抗脱敏治疗DSA阳性患者,检测移植前后DSA水平,主要评估分析其植入情况。结果8例DSA性患者7例获得供者细胞稳定植入,嵌合率均为100%,1例血小板植入不良。经过DFPP、IVIG、利妥昔单抗脱敏处理后为平均荧光强度(mean fluorescence intensity,MFI)(3911±2499),均明显降低,差异均有统计学意义(t=2.101,P<0.001),8例患者中有3例转为弱阳性。干细胞回输第3天复测MFI(907士997),较干细胞回输前再次减低,差异均有统计学意义(t=2.145,P=0.002)。8例患者仅1例发生重度急性移植物抗宿主病。结论双重膜滤过式血浆置换脱敏联合大剂量IVIG和利妥昔单抗,尽量输注高剂量的干细胞,可以降低DSA水平促进供者干细胞植入。

非亲缘供者造血干细胞移植治疗JMML的临床疗效

目的评估非亲缘供者造血干细胞移植(URD-HSCT)治疗幼年粒单核细胞白血病(JMML)的临床疗效及安全性。方法回顾性分析2015年—2023年期间23例JMML移植患儿数据,中位移植年龄33(11-98)个月,PTPN11基因突变为主。供者选择HLA≥9/10位点相合的非亲缘供者。结果中位随访时间31(8-104)个月,输注CD34+中位数为4.8(1.3-12.5)×10^(6)/kg,23例均完全供者植入,粒细胞、血小板植入的中位时间分别为19天和13天,5年OS和LFS的统计预估值均为(91.1±6)%,无复发;Ⅱ度以上急性GVHD 7例(28%),Ⅲ-Ⅳ度仅1例;慢性GVHD 9例(39%),其中重度2例;移植相关死亡率8.9%。结论基于地西他滨的URD-HSCT方案治疗JMML临床疗效显著且安全,可作为JMML移植策略的优先推荐。

免疫组库测序在实体器官移植中的应用

免疫组库一般指T细胞和B细胞的总称,它有着巨大的多样性,使免疫系统能够对多种抗原刺激做出反应。随着测序技术的发展,免疫组库测序可以从基因水平深入了解排斥反应发生时淋巴细胞克隆的变化,也为基于免疫组库测序的新型无创诊断技术的产生提供了可能。近年来,免疫组库测序在实体器官移植中的尝试不断增多,特别是在肾移植、肝移植、心脏移植以及移植后感染等领域。本文对以上领域中应用免疫组库测序的研究进行了综述,总结器官移植中免疫组库测序使用的现状和作为早期无创诊断排斥反应的新技术的潜力,以期为这项技术进一步发展并应用于临床提供借鉴。

供者特异性HLA抗体及去敏治疗对单倍体造血干细胞移植植入效果的影响

目的 分析供者特异性HLA抗体(anti-HLA donor-specific antibodies, DSA)及DSA阳性患者的去敏治疗对单倍体造血干细胞移植(haploidentical hematopoietic stem cell transplantation, haplo-HSCT)植入效果的影响。方法 收集2017年3月至2023年7月重庆医科大学附属第二医院血液内科行haplo-HSCT,并完成HLA抗体及DSA检测的患者70例,分析DSA阳性和DSA阳性患者去敏治疗对造血干细胞移植植入效果的影响。结果 70例haplo-HSCT患者完成抗体检测,DSA阳性患者15例(21.4%),其中7例(46.7%)强阳性,3例(20.0%)中度阳性,5例(33.3%)弱阳性。DSA阳性患者移植后粒系植入中位时间较DSA阴性患者明显延迟(P=0.027)。植入失败(graft failure, GF)患者6例,DSA阳性4例(26.7%),明显高于DSA阴性(P=0.025)。多因素分析结果显示,DSA是发生GF的独立影响因素(HR=9.273, 95%CI=1.505~57.124,P=0.016)。10例DSA中度和强阳性患者进行去敏治疗,4例采用联合去敏治疗,患者均成功植入(100.0%),6例采用单一去敏治疗,有4例(66.7%)发生GF,联合去敏治疗的GF发生率显著低于单一去敏治疗(P=0.008)。结论 DSA是导致haplo-HSCT患者植入延迟和GF的重要因素,对DSA中度及强阳性患者的单一去敏治疗效果有限,而多联合的去敏治疗时,动态监测抗体滴度水平下降,可保证干细胞的成功植入,降低GF发生。

X-ray irradiation selectively kills thymocytes of different stages and impairs the maturation of donor-derived CD4^+CD8^+ thymocytes in recipient thymus

The aim of the present study was to determine whether the sensitivity of thymocytes to X-ray radiation depends on their proliferative states and whether radiation impairs the maturation of donor-derived thymocytes in recipient thymus.We assigned 8-week-old C57BL/6J mice into three treatment groups：1） untreated;2） X-ray radiation;3） X-ray radiation plus bone marrow transplantation with donor bone marrow cells from transgenic mice express-ing enhanced green fluorescent protein（GFP） on a universal promoter.After 4 weeks,the size of the thymus,the number and proliferation of thymocytes and ratios of different stage thymocytes were analyzed by immunohisto-chemistry and flow cytometry.The results showed that：1） CD4＋CD8＋ thymocytes were more sensitive to X-ray radiation-induced cell death than other thymocytes;2） the proliferative capacity of CD4＋CD8＋ thymocytes was higher than that of other thymocytes;3） the size of the thymus,the number of thymocytes and ratios of thymo-cytes of different stages in irradiated mice recovered to the normal level of untreated mice by bone marrow trans-plantation;4） the ratio of GFP-positive CD4＋CD8＋ thymocytes increased significantly,whereas the ratio of GFP-positive CD4＋ or CD8＋ thymocytes decreased significantly.These results indicate that the degree of sensitivity of thymocytes to X-ray radiation depends on their proliferative states and radiation impairs the maturation of donor-derived CD4＋CD8＋ thymocytes in recipient thymus.