Polycomb proteins RING1A/B promote H2A monoubiquitination to regulate female gametophyte development in Arabidopsis.

A functional female gametophyte is the basis of successful sexual reproduction in flowering plants. During female gametophyte development, the megaspore mother cell (MMC), differentiated from a single subepidermal somatic cell in the nucellus, undergoes meiosis to produce four megaspores; only the one at the chalazal end, referred to as functional megaspore (FM), undergoes three rounds of mitosis and develops into a mature embryo sac. Here, we reported that RING1A and RING1B (RING1A/B), two functionally redundant Polycomb proteins in Arabidopsis, are critical for female gametophyte development. The mutations of RING1A/B resulted in defects in MMC and FM's specification and subsequent mitosis of FM, thereby leading to aborted ovules. Gene expression analysis revealed several genes essential for female gametophyte development, including Argonaute (AGO) family genes and critical transcription factors, were ectopically expressed in ring1a ring1b. Furthermore, RING1A/B bound some of these genes to promote H2A monoubiquitination (H2Aub) deposition. Together, RING1A/B promote H2Aub modification at genes essential for female gametophyte development, suppressing their expression to ensure the progression of female gametophyte development.

Persistent Cytopenia After CD19 CAR T Therapy in Relapsed/Refractory DLBCL Patients Could Be a Predictor of Efficacy and Side Effects.

Hematological toxicity is a severe adverse event (AE) in anti-CD19 chimeric antigen receptor (CAR) T cell therapy for relapsed/refractory (R/R) diffuse large B-cell lymphoma (DLBCL). However, the pathophysiological mechanism underlying prolonged cytopenia and the relationship between persistent cytopenia, efficacy, and AEs after anti-CD19 CAR T cell therapy are unknown. Therefore, this study explored whether persistent cytopenia after anti-CD19 CAR T cell therapy in patients with R/R DLBCL can predict therapeutic efficacy and AEs. Thirty-eight patients with R/R DLBCL were enrolled in an anti-CD19 CAR T cell therapy clinical trial. Patients received lymphodepleting chemotherapy with fludarabine and cyclophosphamide before CAR T cell therapy. The degree and duration of cytopenia, clinical response, proportion of CAR T cells, interleukin-6 (IL-6) levels, AEs, and follow-up were observed after therapy. Grades 3-4 persistent cytopenia occurred in 14 patients with R/R DLBCL, who recovered 8-18 weeks after CAR T cell infusion. These patients achieved an objective response rate (ORR) for anti-CD19 CAR T cell therapy. In patients who achieved ORR, the incidence of Grades 3-4 persistent cytopenia was higher in patients with a high tumor load than in those without a high tumor load. The mean peaks of IL-6 and anti-CD19 CAR T cells and the cytokine release syndrome grade in patients with Grades 3-4 persistent cytopenia were higher than those in patients without persistent cytopenia. Anti-CD19 CAR T cells were observed 21 and 28 days after infusion, and patients had Grades 3-4 persistent cytopenia. Progression-free and overall survival were higher in patients with Grades 3-4 persistent cytopenia than in those without cytopenia. Therefore, persistent cytopenia after anti-CD19 CAR T cell therapy in patients with R/R DLBCL can predict therapeutic efficacy and AEs, allowing clinicians to determine the efficiency of CD-19 CAR T cell therapy and the associated AEs.

A Prospective Trial Comparing Haploidentical Donor Transplantation With Cord Blood Versus HLA-Matched Sibling Donor Transplantation for Hematologic Malignancy Patients.

TRIAL REGISTRATION: Effect of co-infusion third party umbilical cord blood stem cells on haploidentical hematopoietic stem cell transplantation https://www.chictr.org.cn Reg. No. ChiCTR-OIN-17011426.

Integrated Analysis of mRNA and Non-coding RNA Transcriptome in Pepper (Capsicum chinense) Hybrid at Seedling and Flowering Stages.

Pepper is an important vegetable in the world. In this work, mRNA and ncRNA transcriptome profiles were applied to understand the heterosis effect on the alteration in the gene expression at the seedling and flowering stages between the hybrid and its parents in Capsicum chinense. Our phenotypic data indicated that the hybrid has dominance in leaf area, plant scope, plant height, and fruit-related traits. Kyoto Encyclopedia of Genes and Genomes analysis showed that nine members of the plant hormone signal transduction pathway were upregulated in the seedling and flowering stages of the hybrid, which was supported by weighted gene coexpression network analysis and that BC332_23046 (auxin response factor 8), BC332_18317 (auxin-responsive protein IAA20), BC332_13398 (ethylene-responsive transcription factor), and BC332_27606 (ethylene-responsive transcription factor WIN1) were candidate hub genes, suggesting the important potential role of the plant hormone signal transduction in pepper heterosis. Furthermore, some transcription factor families, including bHLH, MYB, and HSF were greatly over-dominant. We also identified 2,525 long ncRNAs (lncRNAs), 47 micro RNAs (miRNAs), and 71 circle RNAs (circRNAs) in the hybrid. In particular, downregulation of miR156, miR169, and miR369 in the hybrid suggested their relationship with pepper growth vigor. Moreover, we constructed some lncRNA-miRNA-mRNA regulatory networks that showed a multi-dimension to understand the ncRNA relationship with heterosis. These results will provide guidance for a better understanding of the molecular mechanism involved in pepper heterosis.

Ibrutinib improves the efficacy of anti-CD19-CAR T-cell therapy in patients with refractory non-Hodgkin lymphoma.

The efficacy and side effects of the second-time humanized CD19 chimeric antigen receptor (CD19-CAR) T-cell therapy after unsuccessful first-time anti-CD19-CAR T-cell therapy and subsequent ibrutinib salvage treatment were observed in patients with refractory B-cell lymphoma. In our study, 3 patients with refractory mantle cell lymphoma (MCL) and 4 patients with refractory follicular lymphoma (FL) reached stable disease (SD), partial remission (PR), or progression of disease (PD) after first-time humanized anti-CD19-CAR T-cell therapy. They received ibrutinib as a salvage treatment and kept an SD in the following 7-16 mo, but their disease progressed again during ibrutinib salvage treatment. All 7 patients received a second-time humanized anti-CD19-CAR T-cell therapy, which was the same as their first-time anti-CD19-CAR T-cell therapy. In total, 3 MCL patients and 3 FL patients reached complete response (CR) with the second-time anti-CD19-CAR T-cell therapy combined with ibrutinib, whereas 1 FL patient reached PR. There were no differences in the transduction efficiency and proliferation between the 2 instances of anti-CD19-CAR T-cell therapy. However, the second-time anti-CD19-CAR T-cell therapy led to higher peaks of anti-CD19-CAR T cells and anti-CD19-CAR gene copies, but also to higher grades of cytokine release syndrome (CRS) and more serious hematological toxicity. The successful outcome of the second-time anti-CD19-CAR T-cell therapy might suggest that the previous ibrutinib treatment improved the activities of anti-CD19-CAR T cells.

CD19 CAR-T cell treatment conferred sustained remission in B-ALL patients with minimal residual disease.

The persistence or recurrence of minimal residual disease (MRD) after chemotherapy predicts relapse of B-cell acute lymphoblastic leukemia (B-ALL). CD19-directed chimeric antigen receptor T (CD19 CAR-T) cells have shown promising responses in B-ALL. However, their role in chemotherapy-refractory MRD-positive B-ALL remains unclear. Here we aimed to assess the effectiveness and safety of CD19 CAR-T cells in MRD-positive B-ALL patients. From January 2018, a total of 14 MRD-positive B-ALL patients received one or more infusions of autogenous CD19 CAR-T cells. Among them, 12 patients achieved MRD-negative remission after one cycle of CAR-T infusion. At a median follow-up time of 647 days (range 172-945 days), the 2-year event-free survival rate in MRD-positive patients was 61.2% ± 14.0% and the 2-year overall survival was 78.6 ± 11.0%, which were significantly higher than patients with active disease (blasts ≥ 5% or with extramedullary disease). Moreover, patients with MRD had a lower grade of cytokine release syndrome (CRS) than patients with active disease. However, the peak expansion of CAR-T cells in MRD positive patients showed no statistical difference compared to patients with active disease. Five patients received two or more CAR-T cell infusions and these patients showed a decreased peak expansion of CAR-T cell in subsequent infusions. In conclusion, pre-emptive CD19 CAR-T cell treatment is an effective and safe approach and may confer sustained remission in B-ALL patients with chemotherapy-refractory MRD. The trials were registered at www.chictr.org.cn as ChiCTR-ONN-16009862 (November 14, 2016) and ChiCTR1800015164 (March 11, 2018).

Complete plastome sequence of Euphorbia milii Des Moul. (Euphorbiaceae).

Euphorbia milii (Euphorbiaceae) grows as a scrambling shrub with many branches. Here, we report and characterize the complete plastome of E. milii in an effort to provide genomic resources useful for promoting its systematic research. The plastome of E. milii is found to possess a total length of 160,806 bp with the typical quadripartite structure of angiosperms, contains two Inverted Repeats (IRs) of 26,695 bp, a Large Single-Copy (LSC) region of 90,211 bp and a Small Single-Copy (SSC) region of 17,205 bp. The plastome contains 114 genes, consisting of 80 unique protein-coding genes, 30 unique tRNA genes and four unique rRNA genes. The overall A/T content in the plastome of E. milii is of 64.10%. The phylogenetic analysis indicated that E. milii is close to E. tirucalli within Euphorbiaceae in this study. The complete plastome sequence of E. milii will provide a useful resource for the conservation genetics of this species as well as for the phylogenetic studies of Euphorbiaceae.

[Clinical Effectiveness of Decitabine Combined with Reduced FLAG Regimen in the Treatment of Senile Patients with High-Risk Acute Myeloid Leukemia].

OBJECTIVE: To explore the efficacy and adverse reactions of decitabine combined with reduction FLAG regimen on the senile patients with high-risk AML. METHODS: 12 senile patients with high-risk AML received decitabine combined with reduced FLAG regimen (decitabine 20 mg/m2, intravenous drip, qd, d 1-5; fludarabine 30 mg/m2, intravenous drip lasts 30 min, qd, d 3-6; Ara-C 1 g/m2, intravenous drip, qd, d 3-6; and G-CSF 300 µg/d, subcu- taneous injection, d 2 to neutrophils reached the lowest return to＞1.0×109/L) in our study. The efficacy and adverse reactions of this regimen were analyzed. RESULTS: 9 patients achieved complete remission(CR) after one course of decitabine combined with reduced FLAG regimen, 2 patients achieved partial remission (PR) and 1 patient reached a stable disease (SD). The overall response rate was 92%. The median follow-up period was 7.4 months ranged from 3 to 12 months. The median survival time for all patients was 6.4 months. The main treatment-related toxicities were myelosuppression and infection due to neutropenia. Severe non-hematologic toxicities were not observed in these patients, and there was no treatment-related mortality. CONCLUSION: Decitabine combined with reduced FLAG regimen has a definite clinical efficacy in the treatment of senile patients with high-risk AML. This regimen, as induction remission regimen, can effectively improve the CR rate and reduce the adverse reactions. Therefore, it may be used as one of the preferred induction remission regimen to treat the senile patients with high-risk AML.

Case Report of Anti-CD123 Chimeric Antigen Receptor T-Cell Therapy Followed by Radiotherapy for a Recurrence of Blastic Plasmacytoid Dendritic Cell Neoplasm After Allogeneic Hematopoietic Stem Cell Transplantation.

BACKGROUND: Blastic plasmacytoid dendritic cell neoplasm (BPDCN) is a rare hematopoietic malignancy. There is no standard chemotherapy regimen for BPDCN, and even allogeneic hematopoietic stem cell transplantation (allo-HSCT) has not been able to extend the survival of patients with BPDCN. CASE REPORT: Here, we present a case of recurrence of BPDCN in a patient with new nodules in his head six months after allo-HSCT. He was enrolled in a clinical trial of anti-CD123 chimeric antigen receptor (CAR) T-cell therapy (ChiCTR1900022058). However, there were no significant changes in the nodules 28 days after anti-CD123-CAR T-cell infusion. He received radiotherapy for the nodules when the proportion of anti-CD123-CAR T-cells in the peripheral blood was 2.8% and the adverse events related to the anti-CD123-CAR T-cell therapy were resolved. The proportion of anti-CD123-CAR T-cells, the level of CD123-CAR gene desoxyribonucleic acid, and the serum levels of cytokines in the patient's peripheral blood reached the highest peak 14 days after radiotherapy. Fortunately, the nodules disappeared gradually 28 days after radiotherapy. He achieved complete remission again from the anti-CD123-CAR T-cell therapy followed by radiotherapy. To date, he has maintained progression-free survival with complete donor chimerism for six months after the combination therapy. CONCLUSION: Anti-CD123-CAR T-cell therapy followed by radiotherapy for a recurrence of blastic plasmacytoid dendritic cell neoplasm after allo-HSCT is effective.

The Ectopic Expression of SurvivinT34A and FilC Can Enhance the Oncolytic Effects of Vaccinia Virus in Murine Gastric Cancer.

BACKGROUND/AIMS: Anti-tumor vaccines have been shown to be effective in cancer therapeutics ever since the anti-HPV vaccine was developed. Compared to conventional chemotherapy, anti-tumor vaccines can specifically target cancer cells and they have lower side effects. We developed a recombinant vaccinia virus (VACV) (Western Reserve) WR strain, and we tested its anti-tumor effects in an animal model. METHODS: A recombinant VACV WR strain expressing mutant survivin T34A (SurT34A) and FilC was constructed and validated. Its oncolytic effect was tested in vitro using a CCK-8 assay, and its tolerance and anti-tumor effects were tested in a murine gastric cancer model. The proportion of lymphocytes in the spleen and tumor was determined after antibody-mediated immuno-depletion. RESULTS: The recombinant VACV showed a stronger replication ability in tumor cells, and it was safe in vivo, even at high doses. The combination of vv-SurT34A and vv-FilC resulted in a stronger anti-tumor effect compared to either construct alone. However, the inhibitory effect of vv-SurT34A was stronger than the combination. The recombinant VACV activated the host immune response, as indicated by lymphocyte infiltration in the spleen and tumor tissues. CONCLUSION: The recombinant VACV WR strain expressing SurT34A and FilC is a safe and effective anti-tumor vaccine.

Comparison of outcomes of haploidentical donor hematopoietic stem cell transplantation supported by third-party cord blood with HLA-matched unrelated donor transplantation.

Previous study indicated that co-infusion of cord blood cells may potentially improve the outcome of haploidentical donor (HID) transplantation. In this study, we analyzed the outcomes of patients who underwent HID transplantation supported by cord blood when compared with HLA-matched unrelated donor (URD) transplantation. Starting in 2015, 40 patients with hematopoietic malignancies underwent HID transplantation and 26 patients underwent URD transplantation. Hematopoietic recovery, the incidences of grade II-IV acute graft-versus-host disease (GVHD) and chronic GVHD was comparable in the two groups. At two year, the relapse risk in HID group was significantly lower than in URD group (RR 4.630; 95%CI, 1.081-19.839; p = .039). Moreover, HID group have prolonged PFS (RR 2.642; 95%CI, 1.046-6.672; p = .040). In conclusion, HID transplantation supported by cord blood results in better outcomes compared with URD transplantation and it might be a favorable alternative to a HLA-matched URD transplantation.

Preparation and selective recognition of a novel solid-phase microextraction fiber combined with molecularly imprinted polymers for the extraction of parabens in soy sample.

A prepared molecularly imprinted polymer with ethyl p-hydroxybenzoate as template molecule was applied for the first time to a homemade solid-phase microextraction fiber. The molecularly imprinted polymer-coated solid-phase microextraction fiber was characterized by scanning electron microscopy and thermogravimetric analysis. Various parameters were investigated, including extraction temperature, extraction time, and desorption time. Under the optimum extraction conditions, the molecularly imprinted polymer-coated solid-phase microextraction fiber exhibited higher selectivity with greater extraction capacity toward parabens compared with the nonimprinted polymer-coated solid-phase microextraction fiber and commercial fibers. The molecularly imprinted polymer-coated solid-phase microextraction fiber was tested using gas chromatography to determine parabens, including methyl p-hydroxybenzoate, ethyl p-hydroxybenzoate, and propyl p-hydroxybenzoate. The linear ranges were 0.01-10 µg/mL with a correlation coefficient above 0.9943. The detection limits (under signal-to-noise ratio of 3) were below 0.30 µg/L. The fiber was successfully applied to the simultaneous analysis of three parabens in spiked soy samples with satisfactory recoveries of 95.48, 97.86, and 92.17%, respectively. The relative standard deviations (n=6) were within 2.83-3.91%. The proposed molecularly imprinted polymer-coated solid-phase microextraction method is suitable for selective extraction and determination of trace parabens in food samples.