BACKGROUND Patients with acute-on-chronic liver failure(ACLF)experience severe immune dysfunction.Liver transplantation(LT)significantly improves survival outcomes.However,the characteristics of peripheral blood lymph...BACKGROUND Patients with acute-on-chronic liver failure(ACLF)experience severe immune dysfunction.Liver transplantation(LT)significantly improves survival outcomes.However,the characteristics of peripheral blood lymphocyte subsets(PBLSs)in this patient population are not well defined,and the dynamics of immune reconstitution post-LT are insufficiently understood.AIM To characterize PBLSs in patients with ACLF prior to LT and to evaluate PBLS reconstitution after LT.METHODS Clinical data from patients undergoing LT in the Transplantation Center,The Third Xiangya Hospital from January 2022 to December 2023 were analyzed retrospectively.Our cohort comprised 44 patients with ACLF,16 patients with acute decompensation of cirrhosis,and 23 patients with compensated cirrhosis.Twenty healthy volunteers were included as controls.PBLSs were evaluated across all groups.The relationship between PBLSs and post-LT prognosis was assessed,and dynamic changes in PBLSs among patients with ACLF were analyzed at different time points.RESULTS Patients with ACLF exhibited a marked reduction in PBLSs compared with healthy volunteers.Natural killer(NK)cell counts were further reduced in patients with ACLF when compared with patients with compensated cirrhosis.PBLSs did not correlate with the etiology or severity of ACLF or with established liver failure scores.Following LT,a rapid restoration of NK cells and B cells was observed in patients with ACLF.However,the cluster of differentiation(CD)3+T cell and CD4+T cell counts decreased 14 days post-LT and subsequently returned to preoperative levels by day 21.CONCLUSION Patients with ACLF exhibited markedly reduced PBLSs,with decreased NK cells potentially linked to progression from compensated cirrhosis to liver failure.NK and B cell were rapidly restored after LT.展开更多
Paclitaxel,a tetracyclic diterpenoid,has garnered attention for its potent anti-cancer properties and intricate molecular structure,making it a significant target for chemical synthesis and biosynthesis[1].However,its...Paclitaxel,a tetracyclic diterpenoid,has garnered attention for its potent anti-cancer properties and intricate molecular structure,making it a significant target for chemical synthesis and biosynthesis[1].However,its natural sources are extremely limited,as it is derived exclusively from the bark of endangered genus Taxus plants,which contain paclitaxel in very low concentrations(0.01%−0.05%)[2-3].Recent advances in synthetic biology present promising opportunities to enhance paclitaxel levels in Taxus cell cultures or to enable the reconstitution of its production in heterologous hosts,such as yeast or tobacco(Nicotiana benthamiana).展开更多
The mouse genome has a high degree of homology with the human genome,and its physiological,biochemical,and developmental regulation mechanisms are similar to those of humans;therefore,mice are widely used as experimen...The mouse genome has a high degree of homology with the human genome,and its physiological,biochemical,and developmental regulation mechanisms are similar to those of humans;therefore,mice are widely used as experimental animals.However,it is undeniable that interspecies differences between humans and mice can lead to experimental errors.The differences in the immune system have become an impor-tant factor limiting current immunological research.The application of immunodefi-cient mice provides a possible solution to these problems.By transplanting human immune cells or tissues,such as peripheral blood mononuclear cells or hematopoietic stem cells,into immunodeficient mice,a human immune system can be reconstituted in the mouse body,and the engrafted immune cells can elicit human-specific immune responses.Researchers have been actively exploring the development and differen-tiation conditions of host recipient animals and grafts in order to achieve better im-mune reconstitution.Through genetic engineering methods,immunodeficient mice can be further modified to provide a favorable developmental and differentiation microenvironment for the grafts.From initially only being able to reconstruct single T lymphocyte lineages,it is now possible to reconstruct lymphoid and myeloid cells,providing important research tools for immunology-related studies.In this review,we compare the differences in immune systems of humans and mice,describe the devel-opment history of human immune reconstitution from the perspectives of immuno-deficient mice and grafts,and discuss the latest advances in enhancing the efficiency of human immune cell reconstitution,aiming to provide important references for im-munological related researches.展开更多
Using atomic force microscopy (AFM), the dynamic process of the in vitro nucleosome reconstitution followed by slow dilution from high salt to low salt was visualized. Data showed that the histone octamers were dissoc...Using atomic force microscopy (AFM), the dynamic process of the in vitro nucleosome reconstitution followed by slow dilution from high salt to low salt was visualized. Data showed that the histone octamers were dissociated from DNA at 1M NaCl. When the salt concentration was slowly reduced to 650 mMand 300 mM, the core histones bound to the naked DNA gradually. Once the salt concentration was reduced to 50 mM the classic 'beads-on-a-string' structure was clearly visualized. Furthermore, using the technique of the in vitro reconstitution ofnucleosome,the mono- and di- nucleosomes were assembled in vitro with both HS2core (-10681 to -10970 bp) and NCR2 (-372to -194 bp) DNA sequences in the 5'flanking sequence of human b-globin gene. Data revealed that HMG 1/2 and HMG 14/17 proteins binding to both DNA sequences are changeable following the assembly and disassembly of nucleosomes. We suggest that the changeable binding patterns of HMG 14/17 and HMG1/2 proteins with these regulatory elements may be critical in the process of nucleosome assembly, recruitment of chromatin-modifying activities, and the regulation of human b-globin gene expression.展开更多
Context: Highly active antiretroviral therapy (HAART) inhibits the HIV replication and consequently increases CD4 levels and decreases viral load. This immune system improvement can trigger various immunological pheno...Context: Highly active antiretroviral therapy (HAART) inhibits the HIV replication and consequently increases CD4 levels and decreases viral load. This immune system improvement can trigger various immunological phenomena, entity called Immune Reconstitution Syndrome (IRS). Graves’ disease is a late Immune Reconstitution consequence. Patient: We report the case of a 48 years old man with HIV infection who developed Graves’ disease three years after he was on effective HAART because of the Immune Reconstitution Syndrome. At presentation he had a very low CD4 T-cell count (17 cells/μL). When he started HAART he presented a lipodystrophy syndrome. HAART was changed because of the persistent low CD4-T cells count (less than 100 cell/μL). Afterwards serum lipid levels began to decrease and that was the first manifestation of Graves’ disease, which was diagnosed when CD4 T-cells increased up to 343 cell/μL. Our patient developed Graves’ disease 36 months after initiating effective HAART with protease inhibitors which was coincident with viral suppression and a rise of CD4 T cells. Conclusion: The most immunosuppressed patients with a CD4 T cell count less than 100 cells/μL are at greatest risk for the development of Immune Reconstitution Syndrome after HAART initiation. We conclude that clinicians will have to consider the importance of the early diagnosis of thyroid disease to bring an adequate treatment.展开更多
T cells play a critical role in immunity to protect against pathogens and malignant cells.T cell immunodeficiency is detrimental,especially when T cell perturbation occurs during severe infection,irradiation,chemother...T cells play a critical role in immunity to protect against pathogens and malignant cells.T cell immunodeficiency is detrimental,especially when T cell perturbation occurs during severe infection,irradiation,chemotherapy,and age-related thymic atrophy.Therefore,strategies that enhance T cell reconstitution provide considerable benefit and warrant intensive investigation.Here,we report the construction of a T cell ablation model in Tg(coro1a:Den NTR)zebrafish via metronidazole administration.The nascent T cells are mainly derived from the hematopoietic cells migrated from the kidney,the functional homolog of bone marrow and the complete recovery time is 6.5 days post-treatment.The cxcr4b gene is upregulated in the responsive hematopoietic cells.Functional interference of CXCR4 via both genetic and chemical manipulations does not greatly affect T lymphopoiesis,but delays T cell regeneration by disrupting hematopoietic migration.In contrast,cxcr4b accelerates the replenishment of hematopoietic cells in the thymus.Consistently,Cxcl12b,a ligand of Cxcr4,is increased in the thymic epithelial cells of the injured animals.Decreased or increased expression of Cxcl12b results in compromised or accelerated T cell recovery,respectively,similar to those observed with Cxcr4b.Taken together,our study reveals a role of CXCR4-CXCL12 signaling in promoting T cell recovery and provides a promising target for the treatment of immunodeficiency due to T cell injury.展开更多
This paper describes the operational issues and basic technical requirements of modern aerial photogrammetry. The accuracy of photogrammetric point determination and the y-parallax at corresponding model points is ana...This paper describes the operational issues and basic technical requirements of modern aerial photogrammetry. The accuracy of photogrammetric point determination and the y-parallax at corresponding model points is analyzed when stereo models are reconstituted by using the exterior orientation elements of aerial images. Real aerial photographs, at image scales from 1:2 500 to 1:6 0000, with DGPS/IMU data taken from various topographies in China were processed by our POS-supported bundle block adjustment program WuCAPS. The empirical results verified that the accuracy of the exterior orientation elements from bundle block adjustment meets the requirements of the specifications of topographic mapping. However, the accuracy of the exterior orientation elements determined by POS fails to meet the requirements of the specifica- tions of topographic mapping.展开更多
The development of“mini-guts”organoid originates from the identification of Lgr5+intestinal stem cells(ISCs)and circumambient signalings within their specific niche at the crypt bottom.These in vitro self-renewing“...The development of“mini-guts”organoid originates from the identification of Lgr5+intestinal stem cells(ISCs)and circumambient signalings within their specific niche at the crypt bottom.These in vitro self-renewing“mini-guts”,also named enteroids or colonoids,undergo perpetual proliferation and regulated differentiation,which results in a high-performance,self-assembling and physiological organoid platform in diverse areas of intestinal research and therapy.The triumphant reconstitution of ISC niche in vitro also relies on Matrigel,a heterogeneous sarcoma extract.Despite the promising prospect of organoids research,their expanding applications are hampered by the canonical culture pattern,which reveals limitations such as inaccessible lumen,confine scale,batch to batch variation and low reproducibility.The tumor-origin of Matrigel also raises biosafety concerns in clinical treatment.However,the convergence of breakthroughs in cellular biology and bioengineering contribute to multiform reconstitution of the ISC niche.Herein,we review the recent advances in the microfabrication of intestinal organoids on hydrogel systems.展开更多
Objective To explore the effect of α-galactosyleramide (α-GalCer) on immune reconstitution under acute graft-versus-host disease (aGVHD). Methods BALB/c mice were transplanted with allogeneic C57BL/6 bone marro...Objective To explore the effect of α-galactosyleramide (α-GalCer) on immune reconstitution under acute graft-versus-host disease (aGVHD). Methods BALB/c mice were transplanted with allogeneic C57BL/6 bone marrow cells and splenocytes (both 1 × 10^7) after receiving lethal total-body irradiation, α-GalCer (100 ug/kg) or vehicle (dimethyl- sulfoxide) was administered intraperitoneally immediately after transplantation. The effects of α-GalCer on immune reconstitufion, proliferation of T cells and B cells, hematopoiesis, and thymic microenvironment were assessed. Results The α-GalCer group exhibited higher percentages of CD3^+, CD4^+, CD8^+, B220^+, CD40+, and CD86+cells compared with the vehicle group. The number of colony forming unit per 1000 CD34^+ cells in the et-GalCer group was higher than in the vehicle group (P=0.0012). In vitro proliferation assays showed that the α-GalCer group had higher percentages of CD3^+, CD4^+, CD8^+, and B220^+ cells compared with the vehicle group. As for the results of in vivo proliferation assays, the numbers of CD3^+, CD4^+, CD8^+, and B220^+ cells were higher in the α-GalCer group than in the normal group, especially the number of B220^+ cells (P=0.007). Significant difference was not found in thymocyte count between the α-GalCer group and the vehicle group, nor in the percentages of CD3^+, CD4^+, and CD8^+ cells. Conclusion Administration of tl-GalCer after allogeneic bone marrow transplantation may promote immune reconstitution in the presence of aGVHD.展开更多
During pregnancy, a cellular immunosuppressant status is produced. It is characterized by anti-inflammatory cellular responses that allow embryonic implantation. The reversal of these changes during the postpartum per...During pregnancy, a cellular immunosuppressant status is produced. It is characterized by anti-inflammatory cellular responses that allow embryonic implantation. The reversal of these changes during the postpartum period may result in overt clinical manifestations of otherwise quiescent or latent infections. We present an Acute Respiratory Distress Syndrome (ARDS) case after a caesarean section related to the Immune Reconstitution Inflammatory Syndrome (IRIS) in a Guinean black woman. The patient was treated with large doses of corticosteroids showing a great clinical and radiological improvement.展开更多
The Tunisian Dorsal backland is the Eastern Atlas side of maghrebides. Field data of Fahs area allowed us to develop new interpretations and to characterize the main structural features of the studied devices (Jebel R...The Tunisian Dorsal backland is the Eastern Atlas side of maghrebides. Field data of Fahs area allowed us to develop new interpretations and to characterize the main structural features of the studied devices (Jebel Rouas and Ruissate). Heritage of Zaghouan accident, Triassic salt movements and strike-direction of major synsedimentary faults are the principal causes and results of the skinned and superimposed geometric architecture, generated by the reversed extensional (Jurassic-Cretaceous) tectonics. The actual geometry of Jebel Rouas and Ruissate represents a fault propagation fold, affecting Jurassic and Cretaceous sets. The backland of this thrust fault defines an imbrications structures of Barremian series. Tectonic records activities show the existence of angular unconformities (Oligocene and Eocene series on the Cretaceous sets considered as bedrock), slumps, tectonic breccias and synsedimentary faults are all of them controlled by a deep major accident;N-S to NE-SW and NW-SE. Features of the study area are probably related first;to the blockage of Zaghouan thrust oriented NE-SW in the foreland;then, to the intense halokinetic activity, which facilitates the layers displacement acting as decollment level. The detailed structural and stratigraphic study of Fahs area and its neighbors shows the presence of an intense tangential tectonic during upper Miocene, affecting Meso-Cenozoic sets, because all the structures involved are sealed by Oligocene and Miocene thinned series. This is accentuated by the existence of different sets of decollment at different depths, which are represented by a displacement to the SE through the backland of the Tunisian Dorsal. We define these features as an imbrication and thrusting Out of sequence system.展开更多
A strong gluten wheat cultivar Shannong 12 and a medium-strength wheat cultivar Shannong 11 were used to investigate the effects of wheat protein contents and protein fractions on dough rheological properties using a ...A strong gluten wheat cultivar Shannong 12 and a medium-strength wheat cultivar Shannong 11 were used to investigate the effects of wheat protein contents and protein fractions on dough rheological properties using a reconstitution method. The results indicated that the peak height, peak width, peak integral, resistance to extension and area under the curve were increased when protein content increased to 120, 140, and 160% (w/w) of the protein content in the base flours for doughs made from each wheat cultivar. All protein fractions were added to each of the base flours at three levels (0.25, 0.50 and 1.00%, w/w) based upon the protein content. The mixograph dough development time, peak width, and resistance to extension increased when the glutenin, insoluble glutenin, soluble glutenin, and glutenin macropolymer were added and increased systematically with increasing levels of these fractions. Peak integral increased by adding and increasing protein content, however, albumin-globulin had no obvious effects. Extensibility at rupture decreased when the glutenin, insoluble glutenin, soluble glutenin, and glutenin macropolymer were added, and decreased systematically with increasing levels of these fractions. However, extensibility at rupture increased when the monomeric protein, albumin-globulin, and gliadin were added, and increased systematically with increasing levels of these fractions.展开更多
Summary: The effect of ligustrazine on the expression of CD31 in syngenic bone marrow transplantation (BMT) mice was studied. Fifty-six Balb/c mice were divided into 3 groups: normal control group. BMT control gro...Summary: The effect of ligustrazine on the expression of CD31 in syngenic bone marrow transplantation (BMT) mice was studied. Fifty-six Balb/c mice were divided into 3 groups: normal control group. BMT control group, and ligustrazine treated group. Syngenic BMT mouse models were established according to the literatures. In BMT control group and the ligustrazine treated group, the mice were given respecxively orally 0.2 mL saline and 2 mg ligustrazine twice a day. On the 7th, 14th, and 21st day after BMT, the mice were killed. The expression of CD31 on the surface of bone marrow nuclear cells (BMNC) was detected by flow cytometry. Peripheral blood leukocytes, platelets and BMNC were counted. Histological observation of bone marrow was made. The results showed thai in ligustrazine treated group the peripheral blood leukocylcs, platelets and BMNC counts, and the expression of CD31 on the day 7, 14, 21 after BMT were higher than in BMTcontrol group (P〈0.01 or P〈0.05). In conclusion, ligustrazine could obviously enhance the CD31 expression on the surface of BMNC after syngcnic BMT in mice, which may be one of the mecha- nisms underlying the ligustrazine accelerating hematopoietic reconstitution in syngenic BMT.展开更多
BACKGROUND With the rapid development of haploidentical hematopoietic stem cell transplantation(haplo-HSCT),primary poor graft function(PGF)has become a lifethreatening complication.Effective therapies for PGF are inc...BACKGROUND With the rapid development of haploidentical hematopoietic stem cell transplantation(haplo-HSCT),primary poor graft function(PGF)has become a lifethreatening complication.Effective therapies for PGF are inconclusive.New Chinese patent medicine Pai-Neng-Da(PND)Capsule exerts dual effect in promoting hematopoiesis recovery and regulating immunity.Still,the application of PND capsule in hematopoietic stem cell transplantation,especially in the haplo-HSCT setting,has not yet been reported.AIM To evaluate the role of PND capsule in acute leukemia patients with haplo-HSCT.METHODS We retrospectively collected data of acute leukemia patients who underwent haplo-HSCT at the Affiliated People’s Hospital of Ningbo University between April 1,2015 and June 30,2020.Twenty-nine consecutive patients received oral PND capsule from the sixth day to the first month after haplo-HSCT were included in the PND group.In addition,31 patients who did not receive PND capsule during haplo-HSCT were included in the non-PND group.Subsequently,we compared the therapeutic efficacy according to the western medical evaluation indexes and Chinese medical symptom scores,and the survival between the PND group and the non-PND group,using the chi-square test,Fisher’s exact test,and the Kaplan-Meier method.RESULTS The duration of platelet engraftment was shorter in the PND group than in the non-PND group(P=0.039).The PND group received a lower frequency of red blood cells and platelet transfusions than the non-PND group(P=0.033 and P=0.035,respectively).In addition,PND capsule marginally reduced the rate of PGF(P=0.027)and relapse(P=0.043).After 33(range,4-106)months of follow-up,the 3-year relapse-free survival(P=0.046)and progression-free survival(P=0.049)were improved in the PND group than in the non-PND group.Also,the therapeutic efficacy of the PND group according to Chinese medical symptom scores was significantly better than that of the non-PND group(P=0.022).Moreover,the adverse events caused by PND capsule were mild.Nevertheless,there were no significant differences in the duration of neutrophil engraftment,the risk of infection within 100 days after haplo-HSCT,the acute graft-versus-host disease,or the 3-year overall survival between the two groups.CONCLUSION PND capsule could promote hematopoiesis reconstitution,improve the therapeutic efficacy of Chinese medical symptom scores,present anti-tumor effectiveness,and prolong the survival of acute leukemia patients with haplo-HSCT.展开更多
Total phenolics, anthocyanins and antioxidant capacity of five coloured/pigmented (AR2009-10, Adirondack Red, Adirondack Blue, Congo, and POROIPG22-1) and two unpigmented potato genotypes (“Anuschka” and “Russet Bu...Total phenolics, anthocyanins and antioxidant capacity of five coloured/pigmented (AR2009-10, Adirondack Red, Adirondack Blue, Congo, and POROIPG22-1) and two unpigmented potato genotypes (“Anuschka” and “Russet Burbank”) were assessed in fresh (with and without skin) and commercially processed/cooked/reconstituted products. Ascorbate profiles of the seven genotypes also were investigated using fresh tuber/tissue only. The results showed that genotypes greatly varied in their contents of bioactive compounds. Ascorbate profiles of the genotypes were not associated to any particular flesh colour/pigment. However, the pigmented potatoes had 1.5 to 2.5 times more the phenolics, 2 to 3 times more antioxidant capacity and higher levels of anthocyanins (13.98 to 38.57 mg C3GE-100g FW) compared to unpigmented genotypes. No anthocyanins were detected in the unpigmented potatoes. Significant losses of total phenolics, anthocyanins and total antioxidant capacity were found during peeling (18% - 23%), blanching process (40% - 60%) and further cooking/ reconstitution (7% - 12%) with no prominent genotype differences. Together, 65 to 90% of these bioactive compounds were lost during processing. The results suggested that pigmented potatoes contained higher amounts of total phenolics and anthocyanins and blanching step took away the most of the original bioactive compounds.展开更多
To explore tile effects of ligustrazine on hematopoietic reconstitution and its mechanism after bone marrow transplantation (BMT), the allogenic BMT mice were given intra-abdominal injection of 2,mg ligustrazine twic...To explore tile effects of ligustrazine on hematopoietic reconstitution and its mechanism after bone marrow transplantation (BMT), the allogenic BMT mice were given intra-abdominal injection of 2,mg ligustrazine twice a day. On the 1st, 7th, 14th, and 28th day after BMT, peripheral blood cells and bone marrow nuclear cells (BMNC) were counted, and the histological features were evaluated. On the 7th, 14th, 21st day after BMT, CXCR4 expression on the BMNC was assayed. The results showed that peripheral blood cell counts and BMNC counts in ligustrazine-treated group on the 7th, 14th, 28th day were higher than those in BMT group (P<0. 01 or P<O. 05). The percentage of hematopoietic tissue volume, fat tissue hyperplasia and congestion and dilation degree of microvessel in ligustrazine-treated group on the 7th, 14th, 21st, 28th day was higher than those in BMT group. The CXCR4 expression levels in ligustrazine-treated group were higher than in BMT group (P<0.01 or P<0. 05) on the 7th and 14th day, and were lower than in BMT group on the 21st day (P<0. 01 ). It is concluded that the ligustrazine can accelerate hematopoietic reconstruction, enhance growth of hematopoietic tissues and promote the repair of microvessels. The CXCR4 expres- sion levels on BMNC may be responsible for the effect of ligustrazine.展开更多
Despite continuous progress,the prevention and treatment of human immunodeficiency virus(HIV)/acquired immune deficiency syndrome(AIDS)remain the world’s most serious public health challenges.A key problem is the deg...Despite continuous progress,the prevention and treatment of human immunodeficiency virus(HIV)/acquired immune deficiency syndrome(AIDS)remain the world’s most serious public health challenges.A key problem is the degree of immune function reconstruction after antiretroviral therapy.Antiretroviral therapy has enriched the treatment of HIV/AIDS and improved the present conditions and the life quality of HIV/AIDS patients.However,some patients still fail to achieve normalization of CD4+T lymphocyte counts although persistent virological suppression.These patients are referred to as immunological non-responders,and usually present with severe immunological dysfunction.To date,since the underlying mechanism of incomplete immune reconstitution in HIV/AIDS has not been fully elucidated,remaining to be the focus and difficulties of current research.It is still a challenge to explore a safe,effective,and reliable therapeutic method for immunological non-responders.Due to fewer side effects and lower drug resistance,traditional Chinese medicine is often sought to provide alternative pharmacotherapy for regulating the immunity of immunological non-responders in China.In this review,we aimed at summarizing the latest and most comprehensive information on traditional Chinese medicine therapeutic methods for promoting immune reconstruction.In addition,outlooks and perspectives for possible future research that related are also discussed.展开更多
To observe potential effect of the engineered bone marrow stromal cell line QXMSC1 secreting IL-6 (QXMSCIL-6) on accelerating immune reconstitution in syngeneic bone marrow transplantation in mice, QXMSC1 was transfec...To observe potential effect of the engineered bone marrow stromal cell line QXMSC1 secreting IL-6 (QXMSCIL-6) on accelerating immune reconstitution in syngeneic bone marrow transplantation in mice, QXMSC1 was transfected with the eukaryocytic expression vector pcDNAIL-6, which contained hIL-6 cDNA by liposome-mediated gene transfecting technique. G418-resistance clone was selected by limiting dilution. The highest secreting clone was selected by ELISA assay and used in animal experiments. The recipient mice (BALB/c) were lethally irradiated and cotransplanted syngeneic bone marrow (10 7/mice) and the QXMSC1IL-6 (5×10 5/mice). Lymphocyte proliferation induced by ConA and LPS, helper T lymphocyte precursor (HTLp), cytotoxic T lymphocyte precursor (CTLp), plaque-forming cell (PFC), delayed type hypersensitivity (DTH) were examined 30, 60 days in post transplantation respectively. The results showed that lymphocytes proliferation to ConA and LPS, HTLp, CTLp increased, DTH and PFC were improved by cografted stromal cells QXMSC1IL-6 on 30, 60 days after BMT. These results demonstrated that the bone marrow stromal cell line QXMSC1IL-6 transfected with IL-6 (QXMSC1IL-6) accelerated immune reconstitution in syngeneic bone marrow transplantation.展开更多
Objective To investigate the infection rate of cytomegalovirus(CMV) and Epstein-Barr virus(EBV) in patients with severe aplastic anemia (SAA) after intensive immunosuppressive therapy in combination with a thrombopoie...Objective To investigate the infection rate of cytomegalovirus(CMV) and Epstein-Barr virus(EBV) in patients with severe aplastic anemia (SAA) after intensive immunosuppressive therapy in combination with a thrombopoietin receptor agonist (lST+TPO-RA) as well as assess the clinical impact of treatment.展开更多
基金Supported by the National Natural Science Foundation of China,No.82300857.
文摘BACKGROUND Patients with acute-on-chronic liver failure(ACLF)experience severe immune dysfunction.Liver transplantation(LT)significantly improves survival outcomes.However,the characteristics of peripheral blood lymphocyte subsets(PBLSs)in this patient population are not well defined,and the dynamics of immune reconstitution post-LT are insufficiently understood.AIM To characterize PBLSs in patients with ACLF prior to LT and to evaluate PBLS reconstitution after LT.METHODS Clinical data from patients undergoing LT in the Transplantation Center,The Third Xiangya Hospital from January 2022 to December 2023 were analyzed retrospectively.Our cohort comprised 44 patients with ACLF,16 patients with acute decompensation of cirrhosis,and 23 patients with compensated cirrhosis.Twenty healthy volunteers were included as controls.PBLSs were evaluated across all groups.The relationship between PBLSs and post-LT prognosis was assessed,and dynamic changes in PBLSs among patients with ACLF were analyzed at different time points.RESULTS Patients with ACLF exhibited a marked reduction in PBLSs compared with healthy volunteers.Natural killer(NK)cell counts were further reduced in patients with ACLF when compared with patients with compensated cirrhosis.PBLSs did not correlate with the etiology or severity of ACLF or with established liver failure scores.Following LT,a rapid restoration of NK cells and B cells was observed in patients with ACLF.However,the cluster of differentiation(CD)3+T cell and CD4+T cell counts decreased 14 days post-LT and subsequently returned to preoperative levels by day 21.CONCLUSION Patients with ACLF exhibited markedly reduced PBLSs,with decreased NK cells potentially linked to progression from compensated cirrhosis to liver failure.NK and B cell were rapidly restored after LT.
文摘Paclitaxel,a tetracyclic diterpenoid,has garnered attention for its potent anti-cancer properties and intricate molecular structure,making it a significant target for chemical synthesis and biosynthesis[1].However,its natural sources are extremely limited,as it is derived exclusively from the bark of endangered genus Taxus plants,which contain paclitaxel in very low concentrations(0.01%−0.05%)[2-3].Recent advances in synthetic biology present promising opportunities to enhance paclitaxel levels in Taxus cell cultures or to enable the reconstitution of its production in heterologous hosts,such as yeast or tobacco(Nicotiana benthamiana).
基金Scientific and Technological Resources Coordination Project of Shaanxi Province,Grant/Award Number:2020PT-002,2022PT-43 and CX-PT-18Special Fund for Military Laboratory Animals,Grant/Award Number:SYDW_KY(2021)13State Key Laboratory of Holistic Integrative Management of Gastrointestinal Cancers,Grant/Award Number:CBSKL2022ZZ28。
文摘The mouse genome has a high degree of homology with the human genome,and its physiological,biochemical,and developmental regulation mechanisms are similar to those of humans;therefore,mice are widely used as experimental animals.However,it is undeniable that interspecies differences between humans and mice can lead to experimental errors.The differences in the immune system have become an impor-tant factor limiting current immunological research.The application of immunodefi-cient mice provides a possible solution to these problems.By transplanting human immune cells or tissues,such as peripheral blood mononuclear cells or hematopoietic stem cells,into immunodeficient mice,a human immune system can be reconstituted in the mouse body,and the engrafted immune cells can elicit human-specific immune responses.Researchers have been actively exploring the development and differen-tiation conditions of host recipient animals and grafts in order to achieve better im-mune reconstitution.Through genetic engineering methods,immunodeficient mice can be further modified to provide a favorable developmental and differentiation microenvironment for the grafts.From initially only being able to reconstruct single T lymphocyte lineages,it is now possible to reconstruct lymphoid and myeloid cells,providing important research tools for immunology-related studies.In this review,we compare the differences in immune systems of humans and mice,describe the devel-opment history of human immune reconstitution from the perspectives of immuno-deficient mice and grafts,and discuss the latest advances in enhancing the efficiency of human immune cell reconstitution,aiming to provide important references for im-munological related researches.
基金This work was supported by the National Natural Science Foundation of China(Grant No.39893320 and 39870378)the Foundation of the Chinese Academy of Sciences(Grant No.Kj982-j1-618).
文摘Using atomic force microscopy (AFM), the dynamic process of the in vitro nucleosome reconstitution followed by slow dilution from high salt to low salt was visualized. Data showed that the histone octamers were dissociated from DNA at 1M NaCl. When the salt concentration was slowly reduced to 650 mMand 300 mM, the core histones bound to the naked DNA gradually. Once the salt concentration was reduced to 50 mM the classic 'beads-on-a-string' structure was clearly visualized. Furthermore, using the technique of the in vitro reconstitution ofnucleosome,the mono- and di- nucleosomes were assembled in vitro with both HS2core (-10681 to -10970 bp) and NCR2 (-372to -194 bp) DNA sequences in the 5'flanking sequence of human b-globin gene. Data revealed that HMG 1/2 and HMG 14/17 proteins binding to both DNA sequences are changeable following the assembly and disassembly of nucleosomes. We suggest that the changeable binding patterns of HMG 14/17 and HMG1/2 proteins with these regulatory elements may be critical in the process of nucleosome assembly, recruitment of chromatin-modifying activities, and the regulation of human b-globin gene expression.
文摘Context: Highly active antiretroviral therapy (HAART) inhibits the HIV replication and consequently increases CD4 levels and decreases viral load. This immune system improvement can trigger various immunological phenomena, entity called Immune Reconstitution Syndrome (IRS). Graves’ disease is a late Immune Reconstitution consequence. Patient: We report the case of a 48 years old man with HIV infection who developed Graves’ disease three years after he was on effective HAART because of the Immune Reconstitution Syndrome. At presentation he had a very low CD4 T-cell count (17 cells/μL). When he started HAART he presented a lipodystrophy syndrome. HAART was changed because of the persistent low CD4-T cells count (less than 100 cell/μL). Afterwards serum lipid levels began to decrease and that was the first manifestation of Graves’ disease, which was diagnosed when CD4 T-cells increased up to 343 cell/μL. Our patient developed Graves’ disease 36 months after initiating effective HAART with protease inhibitors which was coincident with viral suppression and a rise of CD4 T cells. Conclusion: The most immunosuppressed patients with a CD4 T cell count less than 100 cells/μL are at greatest risk for the development of Immune Reconstitution Syndrome after HAART initiation. We conclude that clinicians will have to consider the importance of the early diagnosis of thyroid disease to bring an adequate treatment.
基金supported by National Key Research and Development Project(2019YFA802703)National Natural Science Foundation of China Grants(331822033,31771623,32000568)+1 种基金Fundamental Research Funds for the Central Universities Grant(XDJK2020C041)Natural Science Foundation of Chongqing(CSTC2020JCYJMSXMX0104)。
文摘T cells play a critical role in immunity to protect against pathogens and malignant cells.T cell immunodeficiency is detrimental,especially when T cell perturbation occurs during severe infection,irradiation,chemotherapy,and age-related thymic atrophy.Therefore,strategies that enhance T cell reconstitution provide considerable benefit and warrant intensive investigation.Here,we report the construction of a T cell ablation model in Tg(coro1a:Den NTR)zebrafish via metronidazole administration.The nascent T cells are mainly derived from the hematopoietic cells migrated from the kidney,the functional homolog of bone marrow and the complete recovery time is 6.5 days post-treatment.The cxcr4b gene is upregulated in the responsive hematopoietic cells.Functional interference of CXCR4 via both genetic and chemical manipulations does not greatly affect T lymphopoiesis,but delays T cell regeneration by disrupting hematopoietic migration.In contrast,cxcr4b accelerates the replenishment of hematopoietic cells in the thymus.Consistently,Cxcl12b,a ligand of Cxcr4,is increased in the thymic epithelial cells of the injured animals.Decreased or increased expression of Cxcl12b results in compromised or accelerated T cell recovery,respectively,similar to those observed with Cxcr4b.Taken together,our study reveals a role of CXCR4-CXCL12 signaling in promoting T cell recovery and provides a promising target for the treatment of immunodeficiency due to T cell injury.
基金Supported by the National Natural Science Foundation of China (No. 40771176, 40721001), the Program for New Century Excellent Talents in University (No. NCET-04-0662).Acknowledgement The experiment data acquisition are supported by Institute of Remote Sensing Applications Chinese Academy of Sciences, Zhong Fei General Aviation Company, Liaoning Jingwei Surveying & Mapping Technology INC, Dalian Urban Surveying Design Institute, Siwei Aviation Remote Sensing Co. Ltd., Xi'an National surveying & Mapping Aviation Remote Sensing Co. Ltd and so on. These supports are gratefully acknowledged. The author would like to express his hearty gratitude to Fu Jianhong, Xie Chou, Ji Shunping and Yang Ming for participating in partial experiments. The author would like to thank Yang Ming and professor Zhang Jingxiong for their polishing in English.
文摘This paper describes the operational issues and basic technical requirements of modern aerial photogrammetry. The accuracy of photogrammetric point determination and the y-parallax at corresponding model points is analyzed when stereo models are reconstituted by using the exterior orientation elements of aerial images. Real aerial photographs, at image scales from 1:2 500 to 1:6 0000, with DGPS/IMU data taken from various topographies in China were processed by our POS-supported bundle block adjustment program WuCAPS. The empirical results verified that the accuracy of the exterior orientation elements from bundle block adjustment meets the requirements of the specifications of topographic mapping. However, the accuracy of the exterior orientation elements determined by POS fails to meet the requirements of the specifica- tions of topographic mapping.
基金Supported by the 333 High Level Talents Training Project of Jiangsu Province,No.BRA2019011and Project of Military Research,No.CLB19J025 and No.TGKS2019002。
文摘The development of“mini-guts”organoid originates from the identification of Lgr5+intestinal stem cells(ISCs)and circumambient signalings within their specific niche at the crypt bottom.These in vitro self-renewing“mini-guts”,also named enteroids or colonoids,undergo perpetual proliferation and regulated differentiation,which results in a high-performance,self-assembling and physiological organoid platform in diverse areas of intestinal research and therapy.The triumphant reconstitution of ISC niche in vitro also relies on Matrigel,a heterogeneous sarcoma extract.Despite the promising prospect of organoids research,their expanding applications are hampered by the canonical culture pattern,which reveals limitations such as inaccessible lumen,confine scale,batch to batch variation and low reproducibility.The tumor-origin of Matrigel also raises biosafety concerns in clinical treatment.However,the convergence of breakthroughs in cellular biology and bioengineering contribute to multiform reconstitution of the ISC niche.Herein,we review the recent advances in the microfabrication of intestinal organoids on hydrogel systems.
基金Supported by National Natural Science Foundation of China (30670898, 30572108)National Basic Research Program of China (973 Program) (2005CB522400)Capital Research Fund for Medical Development (2007-2040)
文摘Objective To explore the effect of α-galactosyleramide (α-GalCer) on immune reconstitution under acute graft-versus-host disease (aGVHD). Methods BALB/c mice were transplanted with allogeneic C57BL/6 bone marrow cells and splenocytes (both 1 × 10^7) after receiving lethal total-body irradiation, α-GalCer (100 ug/kg) or vehicle (dimethyl- sulfoxide) was administered intraperitoneally immediately after transplantation. The effects of α-GalCer on immune reconstitufion, proliferation of T cells and B cells, hematopoiesis, and thymic microenvironment were assessed. Results The α-GalCer group exhibited higher percentages of CD3^+, CD4^+, CD8^+, B220^+, CD40+, and CD86+cells compared with the vehicle group. The number of colony forming unit per 1000 CD34^+ cells in the et-GalCer group was higher than in the vehicle group (P=0.0012). In vitro proliferation assays showed that the α-GalCer group had higher percentages of CD3^+, CD4^+, CD8^+, and B220^+ cells compared with the vehicle group. As for the results of in vivo proliferation assays, the numbers of CD3^+, CD4^+, CD8^+, and B220^+ cells were higher in the α-GalCer group than in the normal group, especially the number of B220^+ cells (P=0.007). Significant difference was not found in thymocyte count between the α-GalCer group and the vehicle group, nor in the percentages of CD3^+, CD4^+, and CD8^+ cells. Conclusion Administration of tl-GalCer after allogeneic bone marrow transplantation may promote immune reconstitution in the presence of aGVHD.
文摘During pregnancy, a cellular immunosuppressant status is produced. It is characterized by anti-inflammatory cellular responses that allow embryonic implantation. The reversal of these changes during the postpartum period may result in overt clinical manifestations of otherwise quiescent or latent infections. We present an Acute Respiratory Distress Syndrome (ARDS) case after a caesarean section related to the Immune Reconstitution Inflammatory Syndrome (IRIS) in a Guinean black woman. The patient was treated with large doses of corticosteroids showing a great clinical and radiological improvement.
文摘The Tunisian Dorsal backland is the Eastern Atlas side of maghrebides. Field data of Fahs area allowed us to develop new interpretations and to characterize the main structural features of the studied devices (Jebel Rouas and Ruissate). Heritage of Zaghouan accident, Triassic salt movements and strike-direction of major synsedimentary faults are the principal causes and results of the skinned and superimposed geometric architecture, generated by the reversed extensional (Jurassic-Cretaceous) tectonics. The actual geometry of Jebel Rouas and Ruissate represents a fault propagation fold, affecting Jurassic and Cretaceous sets. The backland of this thrust fault defines an imbrications structures of Barremian series. Tectonic records activities show the existence of angular unconformities (Oligocene and Eocene series on the Cretaceous sets considered as bedrock), slumps, tectonic breccias and synsedimentary faults are all of them controlled by a deep major accident;N-S to NE-SW and NW-SE. Features of the study area are probably related first;to the blockage of Zaghouan thrust oriented NE-SW in the foreland;then, to the intense halokinetic activity, which facilitates the layers displacement acting as decollment level. The detailed structural and stratigraphic study of Fahs area and its neighbors shows the presence of an intense tangential tectonic during upper Miocene, affecting Meso-Cenozoic sets, because all the structures involved are sealed by Oligocene and Miocene thinned series. This is accentuated by the existence of different sets of decollment at different depths, which are represented by a displacement to the SE through the backland of the Tunisian Dorsal. We define these features as an imbrication and thrusting Out of sequence system.
基金supported financially by the 863 Program of the Ministry of Science and Technology of China(2006AA10Z1E9)the National Natural Science Foundation of China(30671270)the 863 Key Important Project of the Ministry of Science and Technology,China(2006AA100101).
文摘A strong gluten wheat cultivar Shannong 12 and a medium-strength wheat cultivar Shannong 11 were used to investigate the effects of wheat protein contents and protein fractions on dough rheological properties using a reconstitution method. The results indicated that the peak height, peak width, peak integral, resistance to extension and area under the curve were increased when protein content increased to 120, 140, and 160% (w/w) of the protein content in the base flours for doughs made from each wheat cultivar. All protein fractions were added to each of the base flours at three levels (0.25, 0.50 and 1.00%, w/w) based upon the protein content. The mixograph dough development time, peak width, and resistance to extension increased when the glutenin, insoluble glutenin, soluble glutenin, and glutenin macropolymer were added and increased systematically with increasing levels of these fractions. Peak integral increased by adding and increasing protein content, however, albumin-globulin had no obvious effects. Extensibility at rupture decreased when the glutenin, insoluble glutenin, soluble glutenin, and glutenin macropolymer were added, and decreased systematically with increasing levels of these fractions. However, extensibility at rupture increased when the monomeric protein, albumin-globulin, and gliadin were added, and increased systematically with increasing levels of these fractions.
文摘Summary: The effect of ligustrazine on the expression of CD31 in syngenic bone marrow transplantation (BMT) mice was studied. Fifty-six Balb/c mice were divided into 3 groups: normal control group. BMT control group, and ligustrazine treated group. Syngenic BMT mouse models were established according to the literatures. In BMT control group and the ligustrazine treated group, the mice were given respecxively orally 0.2 mL saline and 2 mg ligustrazine twice a day. On the 7th, 14th, and 21st day after BMT, the mice were killed. The expression of CD31 on the surface of bone marrow nuclear cells (BMNC) was detected by flow cytometry. Peripheral blood leukocytes, platelets and BMNC were counted. Histological observation of bone marrow was made. The results showed thai in ligustrazine treated group the peripheral blood leukocylcs, platelets and BMNC counts, and the expression of CD31 on the day 7, 14, 21 after BMT were higher than in BMTcontrol group (P〈0.01 or P〈0.05). In conclusion, ligustrazine could obviously enhance the CD31 expression on the surface of BMNC after syngcnic BMT in mice, which may be one of the mecha- nisms underlying the ligustrazine accelerating hematopoietic reconstitution in syngenic BMT.
基金Supported by The Zhejiang Provincial Science and Technology Program of Traditional Chinese Medicine,No.2017ZA129 and No.2018ZA112.
文摘BACKGROUND With the rapid development of haploidentical hematopoietic stem cell transplantation(haplo-HSCT),primary poor graft function(PGF)has become a lifethreatening complication.Effective therapies for PGF are inconclusive.New Chinese patent medicine Pai-Neng-Da(PND)Capsule exerts dual effect in promoting hematopoiesis recovery and regulating immunity.Still,the application of PND capsule in hematopoietic stem cell transplantation,especially in the haplo-HSCT setting,has not yet been reported.AIM To evaluate the role of PND capsule in acute leukemia patients with haplo-HSCT.METHODS We retrospectively collected data of acute leukemia patients who underwent haplo-HSCT at the Affiliated People’s Hospital of Ningbo University between April 1,2015 and June 30,2020.Twenty-nine consecutive patients received oral PND capsule from the sixth day to the first month after haplo-HSCT were included in the PND group.In addition,31 patients who did not receive PND capsule during haplo-HSCT were included in the non-PND group.Subsequently,we compared the therapeutic efficacy according to the western medical evaluation indexes and Chinese medical symptom scores,and the survival between the PND group and the non-PND group,using the chi-square test,Fisher’s exact test,and the Kaplan-Meier method.RESULTS The duration of platelet engraftment was shorter in the PND group than in the non-PND group(P=0.039).The PND group received a lower frequency of red blood cells and platelet transfusions than the non-PND group(P=0.033 and P=0.035,respectively).In addition,PND capsule marginally reduced the rate of PGF(P=0.027)and relapse(P=0.043).After 33(range,4-106)months of follow-up,the 3-year relapse-free survival(P=0.046)and progression-free survival(P=0.049)were improved in the PND group than in the non-PND group.Also,the therapeutic efficacy of the PND group according to Chinese medical symptom scores was significantly better than that of the non-PND group(P=0.022).Moreover,the adverse events caused by PND capsule were mild.Nevertheless,there were no significant differences in the duration of neutrophil engraftment,the risk of infection within 100 days after haplo-HSCT,the acute graft-versus-host disease,or the 3-year overall survival between the two groups.CONCLUSION PND capsule could promote hematopoiesis reconstitution,improve the therapeutic efficacy of Chinese medical symptom scores,present anti-tumor effectiveness,and prolong the survival of acute leukemia patients with haplo-HSCT.
文摘Total phenolics, anthocyanins and antioxidant capacity of five coloured/pigmented (AR2009-10, Adirondack Red, Adirondack Blue, Congo, and POROIPG22-1) and two unpigmented potato genotypes (“Anuschka” and “Russet Burbank”) were assessed in fresh (with and without skin) and commercially processed/cooked/reconstituted products. Ascorbate profiles of the seven genotypes also were investigated using fresh tuber/tissue only. The results showed that genotypes greatly varied in their contents of bioactive compounds. Ascorbate profiles of the genotypes were not associated to any particular flesh colour/pigment. However, the pigmented potatoes had 1.5 to 2.5 times more the phenolics, 2 to 3 times more antioxidant capacity and higher levels of anthocyanins (13.98 to 38.57 mg C3GE-100g FW) compared to unpigmented genotypes. No anthocyanins were detected in the unpigmented potatoes. Significant losses of total phenolics, anthocyanins and total antioxidant capacity were found during peeling (18% - 23%), blanching process (40% - 60%) and further cooking/ reconstitution (7% - 12%) with no prominent genotype differences. Together, 65 to 90% of these bioactive compounds were lost during processing. The results suggested that pigmented potatoes contained higher amounts of total phenolics and anthocyanins and blanching step took away the most of the original bioactive compounds.
基金a grant from the NationalNatural Science Fundation of China (No. 39870926).
文摘To explore tile effects of ligustrazine on hematopoietic reconstitution and its mechanism after bone marrow transplantation (BMT), the allogenic BMT mice were given intra-abdominal injection of 2,mg ligustrazine twice a day. On the 1st, 7th, 14th, and 28th day after BMT, peripheral blood cells and bone marrow nuclear cells (BMNC) were counted, and the histological features were evaluated. On the 7th, 14th, 21st day after BMT, CXCR4 expression on the BMNC was assayed. The results showed that peripheral blood cell counts and BMNC counts in ligustrazine-treated group on the 7th, 14th, 28th day were higher than those in BMT group (P<0. 01 or P<O. 05). The percentage of hematopoietic tissue volume, fat tissue hyperplasia and congestion and dilation degree of microvessel in ligustrazine-treated group on the 7th, 14th, 21st, 28th day was higher than those in BMT group. The CXCR4 expression levels in ligustrazine-treated group were higher than in BMT group (P<0.01 or P<0. 05) on the 7th and 14th day, and were lower than in BMT group on the 21st day (P<0. 01 ). It is concluded that the ligustrazine can accelerate hematopoietic reconstruction, enhance growth of hematopoietic tissues and promote the repair of microvessels. The CXCR4 expres- sion levels on BMNC may be responsible for the effect of ligustrazine.
基金the Science and Technology Project of Sichuan Province(No.2020YFS0333).
文摘Despite continuous progress,the prevention and treatment of human immunodeficiency virus(HIV)/acquired immune deficiency syndrome(AIDS)remain the world’s most serious public health challenges.A key problem is the degree of immune function reconstruction after antiretroviral therapy.Antiretroviral therapy has enriched the treatment of HIV/AIDS and improved the present conditions and the life quality of HIV/AIDS patients.However,some patients still fail to achieve normalization of CD4+T lymphocyte counts although persistent virological suppression.These patients are referred to as immunological non-responders,and usually present with severe immunological dysfunction.To date,since the underlying mechanism of incomplete immune reconstitution in HIV/AIDS has not been fully elucidated,remaining to be the focus and difficulties of current research.It is still a challenge to explore a safe,effective,and reliable therapeutic method for immunological non-responders.Due to fewer side effects and lower drug resistance,traditional Chinese medicine is often sought to provide alternative pharmacotherapy for regulating the immunity of immunological non-responders in China.In this review,we aimed at summarizing the latest and most comprehensive information on traditional Chinese medicine therapeutic methods for promoting immune reconstruction.In addition,outlooks and perspectives for possible future research that related are also discussed.
基金This work was supported by a grant from the National Natural Science Foundation of China (No. 30170895)
文摘To observe potential effect of the engineered bone marrow stromal cell line QXMSC1 secreting IL-6 (QXMSCIL-6) on accelerating immune reconstitution in syngeneic bone marrow transplantation in mice, QXMSC1 was transfected with the eukaryocytic expression vector pcDNAIL-6, which contained hIL-6 cDNA by liposome-mediated gene transfecting technique. G418-resistance clone was selected by limiting dilution. The highest secreting clone was selected by ELISA assay and used in animal experiments. The recipient mice (BALB/c) were lethally irradiated and cotransplanted syngeneic bone marrow (10 7/mice) and the QXMSC1IL-6 (5×10 5/mice). Lymphocyte proliferation induced by ConA and LPS, helper T lymphocyte precursor (HTLp), cytotoxic T lymphocyte precursor (CTLp), plaque-forming cell (PFC), delayed type hypersensitivity (DTH) were examined 30, 60 days in post transplantation respectively. The results showed that lymphocytes proliferation to ConA and LPS, HTLp, CTLp increased, DTH and PFC were improved by cografted stromal cells QXMSC1IL-6 on 30, 60 days after BMT. These results demonstrated that the bone marrow stromal cell line QXMSC1IL-6 transfected with IL-6 (QXMSC1IL-6) accelerated immune reconstitution in syngeneic bone marrow transplantation.
文摘Objective To investigate the infection rate of cytomegalovirus(CMV) and Epstein-Barr virus(EBV) in patients with severe aplastic anemia (SAA) after intensive immunosuppressive therapy in combination with a thrombopoietin receptor agonist (lST+TPO-RA) as well as assess the clinical impact of treatment.