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Abstract Background Existing treatments for cholangiocarcinoma have poor efficacy. However, chimeric antigen receptor‐T (CAR‐T) cells are emerging as a potential therapeutic strategy. Solid tumors possess multiple adverse factors in an immunosuppressive microenvironment that impair CAR‐T cell infiltration and function. This study aimed to improve the function of CAR‐T cells through knock down immune checkpoints and immunosuppressive molecular receptors. Methods We evaluated the expression of epidermal growth factor receptor (EGFR) and B7 homolog 3 protein (B7H3) antigens in cholangiocarcinoma tissues using immunohistochemistry and screened specific immune checkpoints in the cholangiocarcinoma microenvironment via flow cytometry. Subsequently, we engineered CAR‐T cells targeting EGFR and B7H3 antigens. We simultaneously knocked down immune checkpoints and immunosuppressive molecular receptors in CAR‐T cells by constructing two clusters of small hairpin RNAs and evaluated the engineered CAR‐T cells for antitumor activity both in vitro, using tumor cell lines and cholangiocarcinoma organoid models, and in vivo, using humanized mouse models. Results We observed high expression of EGFR and B7H3 antigens in cholangiocarcinoma tissues. EGFR‐CAR‐T and B7H3‐CAR‐T cells demonstrated specific anti‐tumor activity. We found an abundance of programmed cell death protein 1 (PD‐1), T cell immunoglobulin and mucin domain‐containing protein 3 (Tim‐3), and T cell immunoglobulin and ITIM domain (Tigit) on infiltrated CD8+ T cells in the cholangiocarcinoma microenvironment. We then decreased the expression of these 3 proteins on the surface of CAR‐T cells, named PTG‐scFV‐CAR‐T cells. Furthermore, we knocked‐down the expression of transforming growth factor beta receptor (TGFβR), interleukin‐10 receptor (IL‐10R), and interleukin‐6 receptor (IL‐6R) of PTG‐scFV‐CAR‐T cells. Those cells, named PTG‐T16R‐scFV‐CAR‐T cells, potently killed tumor cells in vitro and promoted apoptosis of tumor cells in a cholangiocarcinoma organoid model. Finally, the PTG‐T16R‐scFv‐CAR‐T cells showed greater inhibitory effect on tumor growth in vivo, and were superior in prolonging the survival of mice. Conclusions Our results revealed that PTG‐T16R‐scFV‐CAR‐T cells with knockdown of sextuplet inhibitory molecules exhibited strong immunity against cholangiocarcinoma and long‐term efficacy both in vitro and in vivo. This strategy provides an effective and personalized immune cell therapy against cholangiocarcinoma.

Abstract Background Apatinib, a highly selective VEGFR2 inhibitor, significantly improved efficacy versus placebo as a third- and later-line treatment for advanced gastric cancer in phase 2 and 3 trials. This prospective, single-arm, multicenter phase IV AHEAD study was conducted to verify the safety and efficacy of apatinib in patients with advanced or metastatic gastric or gastroesophageal adenocarcinoma after at least two lines of systematic therapy in clinical practice settings. Methods Patients with advanced gastric cancer who had previously failed at least two lines of chemotherapy received oral apatinib until disease progression, death or unacceptable toxicity. The primary endpoint was safety. The secondary endpoints included objective response rate (ORR), disease control rate (DCR), progression-free survival (PFS) and overall survival (OS). Adverse events were summarized by the incidence rate. Median OS and PFS were estimated using the Kaplan–Meier method. ORR, DCR, OS at 3 and 6 months, and PFS at 3 and 6 months were calculated, and their 95% CIs were estimated according to the Clopper-Pearson method. Results Between May 2015 and November 2019, a total of 2004 patients were enrolled, and 1999 patients who received at least one dose of apatinib were assessed for safety. In the safety population, 87.9% of patients experienced treatment-related adverse events (TRAEs), with the most common hypertension (45.2%), proteinuria (26.5%), and white blood cell count decreased (25.3%). Additionally, 51% of patients experienced grade ≥ 3 TRAEs. Fatal TRAEs occurred in 57 (2.9%) patients. No new safety concerns were reported. Among the 2004 patients included in the intention-to-treat population, the ORR was 4.4% (95% CI, 3.6–5.4%), and DCR was 35.8% (95% CI, 33.7–38.0%). The median PFS was 2.7 months (95% CI 2.2–2.8), and the median OS was 5.8 months (95% CI 5.4–6.1). Conclusions The findings in the AHEAD study confirmed the acceptable and manageable safety profile and clinical benefit of apatinib in patients with advanced gastric cancer as a third- or later-line of treatment. Trial registration This study was registered with ClinicalTrials.gov NCT02426034. Registration date was April 24, 2015.

Maize breeding is greatly affected by hybrid vigor, a phenomenon that hybrids exhibit superior performance than parental lines. The immortalized F2 population (IMF2) is ideal for the genetic dissection and prediction of hybrid performance. Here, in this study, we conducted the QTL mapping and genomic prediction of six traits related to plant architecture using an IMF2 population. Broad-sense heritability of these traits ranged from 0.85 to 0.94. Analysis of genetic effects showed that additive variance was the main contributor to phenotypic variations. The mapping of quantitative trait loci (QTLs) revealed 10 to 16 QTLs (including pleiotropic loci and epistatic QTLs) for the six traits. Additionally, we identified 15 fine-tuning QTLs for plant height (PH). For genomic prediction (GP), the model of additive and dominance (AD) exhibited higher prediction accuracy than those fitting general combining ability (GCA) and its combination with special combining ability (SCA) effects for all tested traits. And adding the epistasis (E) effect into the AD model did not significantly increase its prediction accuracy. Moreover, the identified 15 fine-tuning QTLs of PH, which exerted large genomic prediction effects, were verified by the marker effect of GP. Our results not only provide an approach for the fine-mapping of fine-tuning QTLs but also serve as references for GP breeding in crops.

As an important food and forage crops in China, corn plays a key role in guaranteeing food security, promoting social and economic development and providing industrial energy. Cytoplasmic male sterility (CMS) is a common biological phenomenon in higher plants. CMS of maize can be divided into three types: T type, C type and S type. Sterility genes are derived from mitochondrial gene rearrangement to form chimeric genes. The newly formed chimeric genes harm the development of anther microspores and lead to abortion. The existence of restorer gene can eliminate the harm of sterility gene and make microspore grow normally. Since the sterility genes are mitochondrial genes and the restorer genes are nuclear genes, the study on the mechanism of cytoplasmic sterility and restorer is also a bridge to explore the relationship between cytoplasmic and nuclear interactions. At the same time, the utilization of male sterile lines is an important technical means for maize to utilize heterosis. The utilization of cytoplasmic male sterile lines in maize production is of great significance to the utilization of crop heterosis and seed production of hybrid seeds. It can not only liberate labor force and reduce seed production cost, but also improve seed production purity and increase yield. In this paper, the classification and characteristics of CMS in maize as well as the CMS genes and fertility restorer genes discovered in maize in recent years were summarized, and the problems and development prospects in the application of CMS in maize were discussed, which could provide references for the promotion and utilization of CMS in production.

As global industrialization and its associated anthropogenic activities rapidly increase, so too does the areal extent of human-altered soils. The soil classification framework must incorporate the classification schemes of these disturbed soils in order to remediate land. Soil Taxonomy (ST) and the World Reference Base for Soil Resources (WRB) are the most widely used soil classification systems in the world. In this study, 15 typical anthropogenic soil profiles with engineering and technical characteristics from China were selected for a classification study. The aim of this study was to clarify the classifications of these soil profiles in the ST, WRB, Chinese Soil Taxonomy (CST), and Geogenetic Soil Classification of China (GSCC), and make references accordingly. The results showed that the WRB can classify these soils as Technosols in the first level. ST can classify most of these soils as subgroups within the different great groups under Entisols, as well as the Human-Altered and Human-Transported Material classes within the soil family differentiae. For Chinese soil classification schemes, there is a large loophole in the CST regarding the classification of anthropogenic soils. Many anthropogenic soils cannot reflect these soils’ artificial and technical naming characteristics. For the CST, revised proposals based on the WRB and ST were proposed. Based on the artificial disturbance of soil layers, a manufactured layer was added to the diagnostic basis. The Artificalic Anthrosols suborder was added, and it can be divided into the Transporti-Artificalic Anthrosols and Alteri-Artificalic Anthrosols groups. The subgroups were defined by their levels of contamination, imperviousness, and artifact content. The “T” layer symbol was added to soil profile descriptions to reflect a specific layer about technology, such as A, E, B, and C, to reflect the main genetic horizons. This revised classification system is proposed for inclusion in the revised CST to account for the very large and expanding extent of disturbed soils in China and to remain current with other global soil taxonomy systems.

The flat terrain in the plain areas of Beijing, China makes the land easily accessible for cultivation and farming, providing vast opportunities for agricultural development. Meanwhile, these areas are also crucial for urban construction and economic growth. Soil type mapping plays a key role in understanding soil characteristics and guiding land management practices. However, accurately mapping soil types in plain regions can be challenging due to their low spatial variability and diverse land use types. Although land cover changes due to phenomena such as urbanization, agricultural expansion, and conversion of natural vegetation can significantly affect soil properties and distribution patterns, their impacts on soil type mapping remain unclear. This study investigated the impacts of land cover changes in plain areas on the accuracy of soil type mapping, hoping to provide effective assistance for soil type mapping in plain areas by analyzing their coupling relationship. Focusing on the 20 year land cover changes in Tongzhou District, this study utilizes a unified approach that combines expert knowledge, mixed sampling methods, and RF mapping techniques, while incorporating environmental covariates that have minimal period influence and synergistically using NDVI and land cover data from the same year. Transition matrices are used to reveal land cover changes, confusion matrices, and their derived indicators to analyze changes in soil type mapping accuracy, and coupling analysis is conducted between soil type change areas and land cover change areas. The results show that Tongzhou District has experienced rapid development over the past 20 years, with the area of construction land nearly doubling. Additionally, 29% of arable land has been converted into construction land, resulting in an increase in the accuracy of the soil map from 58.99% to 66.91% over the 20 year period. The soil type change area during this period accounts for 16.5% of the total area, with 51.9% of the changed areas overlapping with land cover change areas. These overlapping regions are predominantly influenced by human activities. In terms of cultivated land types in the study area, the quantity of arable land has decreased by approximately 29% over 20 years, while the proportion of Sandy loam calcareous fluvo-aquic soil and Light loam calcareous fluvo-aquic soil, which constitute nearly half of the soil type, has increased. These data demonstrate the coupling relationship between land cover changes and soil type variations. It is evident that improving the extent of land use in plain areas enhances the credibility of soil type mapping. Meanwhile, human activities impact land cover, which, in turn, affects and reflects changes in the soil type.

Prostate cancer (PCa) is the most prevalent cancer among men and the second leading cause of tumor‐associated deaths worldwide, with increasing incidence rates over the last 10 years. Recently, miR‐195 was reported to be hypermethylated at its promoter CpG island and down‐regulated in hepatocellular carcinoma. However, the function of miR‐195 and the underlying mechanisms in PCa remain unknown. Here, we report that a significant down‐regulation of microRNA‐195 (miR‐195) in PCa tissues and cell lines was associated with promoter methylation status. Overexpression of miR‐195 significantly suppressed cell proliferation, migration, invasion and epithelial–mesenchymal transition (increased E‐cadherin and decreased N‐cadherin) in PCa cells. We further demonstrated that transfection with a miR‐195 inhibitor reversed the inhibitory effect of the DNA methyltransferase inhibitor 5‐azacytidine on the proliferation, migration and invasion ability of PCa cells. In summary, our findings suggest that miR‐195 may function as a crucial tumor suppressor in PCa.

The KNOX (KNOTTED1-like homeobox) transcription factors play an important role in leaf, shoot apical meristem and seed development and respond to biotic and abiotic stresses. In this study, we analyzed the diversity and evolutionary history of the KNOX gene family in the genome of tetraploid cotton (Gossypium hirsutum). Forty-four putative KNOX genes were identified. All KNOX genes from seven higher plant species were classified into KNOXI, KNOXII, and KNATM clades based on a phylogenetic analysis. Chromosomal localization and collinearity analysis suggested that whole-genome duplication and a polyploidization event contributed to the expansion of the cotton KNOX gene family. Analyses of expression profiles revealed that the GhKNOX genes likely responded to diverse stresses and were involved in cotton growth developmental processes. Silencing of GhKNOX2 enhanced the salt tolerance of cotton seedlings, whereas silencing of GhKNOX10 and GhKNOX14 reduced seedling tolerance to salt stress. Silencing of GhSTM3 influenced the cotton flowering time and plant development. These findings clarify the evolution of the cotton KNOX gene family and provide a foundation for future functional studies of KNOX proteins in cotton growth and development and response to abiotic stresses.

China is likely to lead global offshore wind power development, in the hope of transforming the coal-based electricity system and reducing greenhouse gas emissions. However, the potential of power generation and emissions mitigation is largely unknown, and the contribution of offshore wind utilization to regional carbon neutrality needs to be further clarified. Here, we reveal that offshore wind energy resources are abundant in China, with an estimated power generation potential of about 17.5 PWh, more than doubling the current power consumption nationwide. Although current utilization of offshore wind energy in China accounts for 21% of global overall capacity, the total share is still limited, supplying just 0.4% of national electricity needs (2019). With the increasing use of offshore wind, by 2050, the planned installation along China coast would be nearly five times as much as current (2019) global capacity, or 25 times of current national offshore wind power generation. The total CO2 emissions reduction in 2050 due to the decrease in coal use is projected to be 294.3 Tg CO2 -eq yr-1, equivalent to 20% of current emissions from coal-fired power in the coastal region. The size of reduced emissions is higher than current CO2 emissions in about 90% of countries. Our results highlight the important role of offshore wind power in upgrading the energy system and achieving carbon neutrality. Future studies are encouraged to further explore technological, economic and institutional challenges facing offshore wind energy deployment and low-carbon energy system development. [ABSTRACT FROM AUTHOR]

Purpose Rituximab with cyclophosphamide, doxorubicin, vincristine, and prednisone administered every 3 weeks (R-CHOP-21) is the standard care for diffuse large B-cell lymphoma (DLBCL). It is unknown whether the dose-dense R-CHOP (R-CHOP-14) could improve the outcome of the disease in Asian population. Materials and Methods Newly diagnosed DLBCL patients were centrally, randomly assigned (1:1) to receive R-CHOP- 14 or R-CHOP-21. R-CHOP-14 was administered every 2 weeks, and R-CHOP-21 was administered every 3 weeks. Primary end point was disease-free survival (DFS). Secondary end points included overall survival (OS), progression-free survival (PFS), response rate and toxicities. Results Seven hundred and two patients were randomly assigned to receive R-CHOP-14 (n=349) or R-CHOP-21 (n=353). With a median follow-up of 45.6 months, the two groups did not differ significantly in 3-year DFS (79.6% for R-CHOP-14 vs. 83.2% for R-CHOP-21, p=0.311), 3-year OS (77.5% for R-CHOP-14 vs. 77.6% for R-CHOP-21, p=0.903), or 3-year PFS (63.2% for R-CHOP-14 vs. 66.1% for R-CHOP-21, p=0.447). Patients with an International Prognostic Index (IPI) score ! 2 had a poorer prognosis compared to those with an IPI score < 2. Grade 3/4 hematologic and non-hematologic toxicities were manageable and similar between R-CHOP-14 and R-CHOP-21. Conclusion R-CHOP-14 did not improve the outcome of DLBCL compared to R-CHOP-21 in Asian population. With manageable and similar toxicities, both of the two regimens were suitable for Asian DLBCL patients. For high-risk patients with IPI ≥ 2, new combination regimens based on R-CHOP deserve further investigation to improve efficacy. [ABSTRACT FROM AUTHOR]

Background: Maize grain yield depends mainly on the photosynthetic efficiency of functional leaves, which is controlled by an array of gene networks and other factors, including environmental conditions. MicroRNAs (miRNAs) are small RNA molecules that play important roles in plant developmental regulation. A few senescenceassociated miRNAs (SA-miRNAs) have been identified as important participants in regulating leaf senescence by modulating the expression levels of their target genes. Results: To elucidate miRNA roles in leaf senescence and their underlying molecular mechanisms in maize, a staygreen line, Yu87-1, and an early leaf senescence line, Early leaf senescence-1 (ELS-1), were selected as experimental materials for the differential expression of candidate miRNAs. Four small RNA libraries were constructed from ear leaves at 20 and 30 days after pollination and sequenced by Illumina deep sequencing technology. Altogether, 81 miRNAs were detected in both lines. Of these, 16 miRNAs of nine families were differentially expressed between ELS-1 andYu87-1. The phenotypic and chlorophyll content analyses of both lines identified these 16 differentially expressed miRNAs as candidate SA-miRNAs. Conclusions: In this study, 16 candidate SA-miRNAs of ELS-1 were identified through small RNA deep sequencing technology. Degradome sequencing results indicated that these candidate SA-miRNAs may regulate leaf senescence through their target genes, mainly transcription factors, and potentially control chlorophyll degradation pathways. The results highlight the regulatory roles of miRNAs during leaf senescence in maize. [ABSTRACT FROM AUTHOR]

IMPORTANCE Hepatitis B virus (HBV) reactivation is a serious complication for patients with lymphoma treated with rituximab-containing chemotherapies, despite lamivudine prophylaxis treatment. An optimal prophylactic antiviral protocol has not been determined. OBJECTIVE To compare the efficacy of entecavir and lamivudine in preventing HBV reactivation in patients seropositive for the hepatitis B surface antigen with untreated diffuse large B-cell lymphoma receiving chemotherapy treatment with rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP). DESIGN, SETTING, AND PATIENTS Randomized, open-label, phase 3 study conducted from February 2008 through December 2012 at 10 medical centers in China. This study was a substudy of a parent study designed to compare a 3-week with a 2-week R-CHOP chemotherapy regimen for untreated diffuse large B-cell lymphoma. Patients enrolled in the parent study who were seropositive for the hepatitis B surface antigen and had normal liver function, serum HBV DNA levels of less than 103 copies/mL, and no prior antiviral therapy were randomized to entecavir (n = 61) or lamivudine (n = 60). INTERVENTIONS Daily entecavir (0.5mg) or lamivudine (100mg) beginning 1 week before the initiation of R-CHOP treatment to 6 months after completion of chemotherapy. MAIN OUTCOMES AND MEASURES The primary efficacy end pointwas the incidence of HBV-related hepatitis. The secondary end points included rates of HBV reactivation, chemotherapy disruption due to hepatitis, and treatment-related adverse events. RESULTS The date of last patient follow-up wasMay 25, 2013. Incidence of HBV-related hepatitis was significantly lower for the entecavir group vs the lamivudine group. CONCLUSIONS AND RELEVANCE Among patients seropositive for the hepatitis B surface antigen with diffuse large B-cell lymphoma undergoing R-CHOP chemotherapy, the addition of entecavir compared with lamivudine resulted in a lower incidence of HBV-related hepatitis and HBV reactivation. If replicated, these findings support the use of entecavir in these patients. [ABSTRACT FROM AUTHOR]

Peroxisome proliferator-activated receptor-gamma (PPAR-γ) is one of the best characterized nuclear hormone receptors (NHRs) in the superfamily of ligand-activated transcriptional factors. PPAR-γ ligands have recently been demonstrated to affect proliferation, differentiation and apoptosis of different cell types. The present study was undertaken to investigate PPAR-γ ligands induced cell growth inhibition and its influence on matrix metalloproteinase MMP-9 and MMP-2 activities on leukemia K562 and HL-60 cells in vitro. The results revealed that PPAR-γ expression was detectable in the two kinds of leukemia cells; Both 15-deoxy-delta(12,14)-prostaglandin J2(15d-PGJ2) and troglitazone (TGZ) have significant growth inhibition effects on these two kinds of leukemia cells. These two PPAR-γ ligands could inhibit the leukemic cell adhesion to the extracellular matrix (ECM) proteins and the invasion through matrigel matrix. The expressions of MMP-9 and MMP-2 as well as their gelatinolytic activities in both HL-60 and K562 cells were inhibited by 15d-PGJ2 and TGZ significantly. We therefore conclude that PPAR-γ ligands 15d-PGJ2 and TGZ have significant growth inhibition effects on myeloid leukemia cells in vitro, and that PPAR-γ ligands can inhibit K562 and HL-60 cell adhesion to and invasion through ECM as well as downregulate MMP-9 and MMP-2 expressions. The data suggest that PPAR-γ ligands may serve as potential anti-leukemia reagents. [ABSTRACT FROM AUTHOR]

The anti-proliferation effects of oridonin on acute promyelocytic leukemia (APL) cells and its mechanisms were studied in vitro. NB4 cells as well as fresh leukemia cells obtained from APL patients in culture medium were treated with different concentrations of oridonin. Cell growth inhibition, apoptosis and related pathways were assessed by MTT assay as well as flow cytometry (FCM) and western blot analysis. The data revealed that oridonin (over 16?µmol/L) could inhibit the growth of NB4 cells by induction of apoptosis. Marked changes of cell apoptosis were observed very clearly by using electron microscopy and DNA fragmentation analysis after the cells exposed to oridonin for 48?h; Western blotting showed cleavage of the caspase-3 zymogen protein (32-kDa) with the appearance of its 20-kDa subunit as well as a cleaved 89-kDa fragment of 116-kDa PARP when apoptosis occurred. The expression of Bcl-2 was down-regulated remarkably accompanied by the disruption of the mitochondrial membrane potential (??m). The anti-proliferative and apoptosis-inducing effects by oridonin in fresh APL cells were also found remarkably using Trypan Blue dye exclusion method and Wright's staining. We concluded that oridoning has significant anti-proliferative and apoptosis-inducing effects on NB4 cells by activation of caspase-3 and cleavage of PARP as well as by down regulation of Bcl-2 and disruption of the ??m. Furthermore, oridonin demonstrated apparent cell growth inhibition effects on fresh APL cells in vitro. The results indicated that oridonin may serve as a potential anti-leukemia reagent. [ABSTRACT FROM AUTHOR]

Jurkat cells in culture medium in vitro were exposed to different concentrations of oridonin. The proliferation rate of the cells was measured by MTT assay, cell apoptotic rate was detected by flow cytometry, morphology of cell apoptosis was observed by Hoechst 33258 fluorescence staining, DNA fragmentation was assayed by agarose gel electrophoresis, caspase-3 and poly(ADP-ribose) polymerase (PARP) expressions were detected by Western blotting using polyclonal anti-caspase-3 antibody and mouse anti-human PARP monoclonal antibodies, and caspase-3 activity was assayed with a colorimetric assay kit before and after apoptosis had occurred. Oridonin (over 32 µmol/l) inhibited the growth of Jurkat cells and caused significant apoptosis. The suppression was both time-dependent and dose-dependent. Marked morphological changes of cell apoptosis, including condensation of chromatin and nuclear fragmentation, were clearly observed by Hoechst 33258 fluorescence staining, as well as by agarose gel electrophoresis. Western blotting showed cleavage of the caspase-3 zymogen protein (32 kDa), with the appearance of its 17 kDa subunit, and a cleaved 89-kDa fragment of 116 kDa PARP was also found, together with a concurrent increase in caspase-3 apoptotic activity. Oridonin can induce apoptosis in Jurkat cells via activation of caspase-3; the results indicate that oridonin might be an important potential anti-leukaemia reagent. [ABSTRACT FROM AUTHOR]

The article provides information on a study which investigated the mechanism by which tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) induces glioma cell apoptosis. The study's objective was to answer the question as to whether the mitochondrial pathway is involved in TRAIL-induced glioma cell apoptosis. The study used human recombinant soluble TRAIL (hrsTRAIL) expressed in Escherichia coli to treat in vitro cultured human glioma U87 cells.