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Mitochondrial DNA (mtDNA) G-quadruplexes (G4s) have important regulatory roles in energy metabolism, yet their specific functions and underlying regulatory mechanisms have not been delineated. Using a chemical-genetic screening strategy, we demonstrated that the JAK/STAT3 pathway is the primary regulatory mechanism governing mtDNA G4 dynamics in hypoxic cancer cells. Further proteomic analysis showed that activation of the JAK/STAT3 pathway facilitates the translocation of RelA, a member of the NF-κB family, to the mitochondria, where RelA binds to mtDNA G4s and promotes their folding, resulting in increased mtDNA instability, inhibited mtDNA transcription, and subsequent mitochondrial dysfunction. This binding event disrupts the equilibrium of energy metabolism, catalyzing a metabolic shift favoring glycolysis. Collectively, the results provide insights into a strategy employed by cancer cells to adapt to hypoxia through metabolic reprogramming., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Phytoremediation largely involves microbial degradation of organic pollutants in rhizosphere for removing organic pollutants like polycyclic aromatic hydrocarbons, phthalates and polychlorinated biphenyls. Microbial community in rhizosphere experiences complex processes of response-adaptation-feedback up on exposure to organic pollutants. This review summarizes recent research on the response and adaptation of rhizosphere microbial community to the stress of organic pollutants, and discusses the enrichment of the pollutant-degrading microbial community and genes in the rhizosphere for promoting bioremediation. Soil pollution by organic contaminants often reduces the diversity of rhizosphere microbial community, and changes its functions. Responses vary among rhizosphere microbiomes up on different classes of organic pollutants (including co-contamination with heavy metals), plant species, root-associated niches (e.g., rhizosphere, rhizoplane and endosphere), geographical location and soil properties. Soil pollution can deplete some sensitive microbial taxa and enrich some tolerant microbial taxa in rhizosphere. Furthermore, rhizosphere enriches pollutant-degrading microbial community and functional genes including different gene clusters responsible for biodegradation of organic pollutants and their intermediates, which improve the adaptation of microbiome and enhance the remediation efficiency of the polluted soil. The knowledge gaps and future research challenges are highlighted on rhizosphere microbiome in response-adaptation-feedback processes to organic pollution and rhizoremediation. This review will hopefully update understanding on response-adaptation-feedback processes of rhizosphere microbiomes and rhizoremediation for the soil with organic pollutants., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Mitochondrion-lysosome interactions have garnered significant attention in recent research. Numerous studies have shown that mitochondrion-lysosome interactions, including mitochondrion-lysosome contact (MLC) and mitophagy, are involved in various biological processes and pathological conditions. Single fluorescent probes are termed a pivotal chemical tool in unraveling the intricate spatiotemporal interorganelle interplay in live cells. However, current chemical tools are insufficient to deeply understand mitochondrion-lysosome dynamic interactions and related diseases, Moreover, the rational design of mitochondrion-lysosome dual-targeting fluorescent probes is intractable. Herein, we designed and synthesized a pH-sensitive fluorescent probe called INSA , which could simultaneously light up mitochondria (red emission) and lysosomes (green emission) for their internal pH differences. Employing INSA , we successfully recorded long-term dynamic interactions between lysosomes and mitochondria. More importantly, the increasing mitochondrion-lysosome interactions in ferroptotic cells were also revealed by INSA . Further, we observed pH variations in mitochondria and lysosomes during ferroptosis for the first time. In brief, this work not only introduced a pH-sensitive fluorescent probe INSA for the disclosure of the mitochondrion-lysosome dynamic interplays but also pioneered the visualization of the organellar pH alternation in a specific disease model.

Endophytic bacteria can degrade toxic phthalate (PAEs). Nevertheless, the colonization and function of endophytic PAE-degrader in soil-crop system and their association mechanism with indigenous bacteria in PAE removal remain unknown. Here, endophytic PAE-degrader Bacillus subtilis N-1 was marked with green fluorescent protein gene. Inoculated strain N-1-gfp could well colonize in soil and rice plant exposed to di-n-butyl phthalate (DBP) as directly confirmed by confocal laser scanning microscopy and realtime PCR. Illumina high-throughput sequencing demonstrated that inoculated N-1-gfp shifted indigenous bacterial community in rhizosphere and endosphere of rice plants with significant increasing relative abundance of its affiliating genus Bacillus than non-inoculation. Strain N-1-gfp exhibited efficient DBP degradation with 99.7% removal in culture solutions, and significantly promoted DBP removal in soil-plant system. Strain N-1-gfp colonization help plant enrich specific functional bacteria (e.g., pollutant-degrading bacteria) with significant higher relative abundances and stimulated bacterial activities (e.g., pollutant degradation) compared with non-inoculation. Furthermore, strain N-1-gfp displayed strong interaction with indigenous bacteria for accelerating DBP degradation in soil, decreasing DBP accumulation in plants and promoting plant growth. This is the first report on well colonization of endophytic DBP-degrader Bacillus subtilis in soil-plant system and its bioaugmentation with indigenous bacteria for promoting DBP removal., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Developing c-MYC transcription inhibitors that target the G-quadruplex has generated significant interest; however, few compounds have demonstrated specificity for c-MYC G-quadruplex and cancer cells. In this study, we designed and synthesized a series of benzoazole derivatives as potential G-quadruplex ligand-based c-MYC transcription inhibitors. Surprisingly, benzoselenazole derivatives, which are rarely reported as G-quadruplex ligands, demonstrated greater c-MYC G-quadruplex selectivity and cancer cell specificity compared to their benzothiazole and benzoxazole analogues. The most promising compound, benzoselenazole m-Se3 , selectively inhibited c-MYC transcription by specifically stabilizing the c-MYC G-quadruplex. This led to selective inhibition of hepatoma cell growth and proliferation by affecting the MYC target gene network, as well as effective tumor growth inhibition in hepatoma xenografts. Collectively, our study demonstrates that m-Se3 holds significant promise as a potent and selective inhibitor of c-MYC transcription for cancer treatment. Furthermore, our findings inspire the development of novel selenium-containing heterocyclic compounds as c-MYC G-quadruplex-specific ligands and transcription inhibitors.

Mutations in NRAS promote tumorigenesis and drug resistance. As this protein is often considered an undruggable target, it is urgent to develop novel strategies to suppress NRAS for anticancer therapy. Recent reports indicated that a G-quadruplex (G4) structure formed in the untranslated region of NRAS mRNA can downregulate NRAS translation, suggesting a potential NRAS suppression strategy. Here, we developed a novel cell-based method for large-scale screening of NRAS G4 ligand using the G-quadruplex-triggered fluorogenic hybridization probe and successfully identified the clinically used agent Octenidine as a potent NRAS repressor. This compound suppressed NRAS translation, blocked the MAPK and PI3K-AKT signaling, and caused concomitant cell cycle arrest, apoptosis, and autophagy. It exhibited better antiproliferation effects over clinical antimelanoma agents and could inhibit the growth of NRAS -mutant melanoma in a xenograft mouse model. Our results suggest that Octenidine may be a prominent anti- NRAS -mutant melanoma agent and represent a new NRAS -mutant melanoma therapy option.

Inter-organelle interactions play a vital role in diverse biological processes. Thus, chemical tools are highly desirable for understanding the spatiotemporal dynamic interplay among organelles in live cells and in vivo . However, designing such tools is still a great challenge due to the lack of universal design strategies. To break this bottleneck, herein, a novel unimolecular platform integrating the twisted intramolecular charge transfer (TICT) and aggregation-induced emission (AIE) dual mechanisms was proposed. As a proof of concept, two organelles, lipid droplets (LDs) and mitochondria, were selected as models. Also, the first TICT-AIE integration molecule, BETA-1 , was designed for simultaneous and dual-color imaging of LDs and mitochondria. BETA-1 can simultaneously target LDs and mitochondria due to its lipophilicity and cationic structure and emit cyan fluorescence in LDs and red fluorescence in mitochondria. Using BETA-1 , for the first time, we obtained long-term tracking of dynamic LD-mitochondrion interactions and identified several impressive types of dynamic interactions between these two organelles. More importantly, the increase in LD-mitochondrion interactions during ferroptosis was revealed with BETA-1 , suggesting that intervening in the LD-mitochondrion interactions may modulate this cell death. BETA-1 was also successfully applied for in vivo imaging of LD-mitochondrion interactions in C. elegans . This study not only provides an effective tool for uncovering LD-mitochondrion interactions and deciphering related biological processes but also sheds light on the design of new probes with an integrated TICT-AIE mechanism for imaging of inter-organelle interactions.

RNA imaging is of great importance for understanding its complex spatiotemporal dynamics and cellular functions. Considerable effort has been devoted to the development of small-molecule fluorescent probes for RNA imaging. However, most of the reported studies have mainly focused on improving the photostability, permeability, long emission wavelength, and compatibility with live-cell imaging of RNA probes. Less attention has been paid to the selectivity and detection limit of this class of probes. Highly selective and sensitive RNA probes are still rarely available. In this study, a new set of styryl probes were designed and synthesized, with the aim of upgrading the detection limit and maintaining the selectivity of a lead probe QUID-1 for RNA. Among these newly synthesized compounds, QUID-2 was the most promising candidate. The limit of detection (LOD) value of QUID-2 for the RNA was up to 1.8 ng/mL in solution. This property was significantly improved in comparison with that of QUID-1 . Further spectroscopy and cell imaging studies demonstrated the advantages of QUID-2 over a commercially available RNA staining probe, SYTO RNASelect, for highly selective and sensitive RNA imaging. In addition, QUID-2 exhibited excellent photostability and low cytotoxicity. Using QUID-2 , the global dynamics of RNA were revealed in live cells. More importantly, QUID-2 was found to be potentially applicable for detecting RNA granules in live cells. Collectively, our work provides an ideal probe for RNA imaging. We anticipate that this powerful tool may create new opportunities to investigate the underlying roles of RNA and RNA granules in live cells.

The research and development of penicillin started with difficulty before 1949 and achieved certain results. In 1951, after the founding of the People's Republic of China, Zhang Weishen, as the only Chinese scientist who had been trained and worked in a penicillin research and development center in the United States for many years, overcame many difficulties and returned to China. In 1953, with the efforts of Zhang Weishen and his colleagues, China realized the industrialized production of penicillin, alleviating the urgent needs of the masses. Antibiotics has also become the first discipline to achieve major scientific and technological achievements after the founding of the New China. In the mid-1950s, the technical breakthrough in the localization of lactose substitutes marked the localization of the raw materials of the penicillin-producing culture medium, which paved the way for the industrialized production of penicillin with Chinese characteristics. Antibiotics have become one of the most widely used and affordable drugs for hundreds of millions of patients in China, and China has since ended the humiliating history of the "Sick Man of East Asia".

Background: Talaromyces marneffei ( T . marneffei ) is an opportunistic fungal pathogen commonly found in human immunodeficiency virus (HIV) patients that often infects lymph nodes. Knowledge about the computed tomography (CT) characteristics of T . marneffei lymphadenopathy in HIV patients is limited. The aim of this study was to investigate the clinical and CT characteristics of T . marneffei lymphadenopathy to improve its diagnosis and promote recognition of this type of infection in radiology., Methods: Between February 2019 and June 2021, we retrospectively reviewed the clinical features and CT characteristics of T . marneffei lymphadenopathy in 21 HIV patients., Results: The clinical symptoms of T . marneffei infection are non-specific. Anemia (100%), fever (85.7%) and cough and sputum production (76.2%) were the most frequent symptoms. Multiple lymphadenopathies, mainly in the mediastinum (76.2%) and mesentery (82.4%), can be fused (14.3%) and necrotic (52.4%), with slight (41.7%) and moderate enhancement (58.3%) that is heterogeneous. In addition to involving the lymph nodes, the lesions involved the lungs (81.0%), liver and spleen (42.9%), and small intestine (14.3%)., Conclusions: T . marneffei is prone to affecting lymph nodes and extranodal organs in HIV patients. Although the clinical manifestations of T . marneffei infection are not specific, the possibility of T. marneffei infection should be considered if CT findings indicate multiple lesion sites., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Zhu, Tang, Liu, Li, Lv and Wang.)

Objective: To explore the effect of CXC chemokine CXCL10 in the mice with diabetic retinopathy (DR)., Methods: DR models were constructed on mice via injection of streptozotocin (STZ). At 3 weeks of STZ, mice were treated with anti-CXCL10 monoclonal antibodies (mAb)/control mAb, and a series of experiments were then conducted at 6 weeks of STZ, including HE staining, western blotting, retinal trypsin digestion, real-time qPCR and enzyme-linked immuno sorbent assay (ELISA). The corresponding kits were used to detect the activity of oxidative stress markers., Results: Compared with nondiabetic eyes, DR mice both at 3 and 6 weeks presented the decreases in the total retinal thickness, the retinal outer nuclear layer (ONL) thickness, and the cells of ganglion cell layer (GCL), with the upregulated CXCL10, pro-inflammatory cytokines and malondialdehyde (MDA), as well as the downregulated levels of superoxide dismutase (SOD), glutathione peroxidase (GPx) and CAT, especially in those at 6 weeks, which were attenuated by the anti-CXCL10 mAb treatment. Moreover, in comparison with the DR mice, mice in the DR + anti-CXCL10 mAb group gained the significant decrease in the number of acellular capillaries of retina, with up-regulations of Claudin-5 and ZO-1 and down-regulations of VEGF and FGF-2. The DR mice injected with anti-CXCL10 mAb demonstrated alleviated retinal pathology as compared to mice at 3 weeks of STZ., Conclusion: Anti-CXCL10 mAb could mitigate the retinal pathology of DR mice, with the decreased inflammation and oxidative stress, thus mediating a delay in the development or disease improvement in patients of DR., (Copyright © 2022. Published by Elsevier Ltd.)

Objective: To investigate the prevalence and genetic variation of Theileria in yellow cattle in Xiangxi Autonomous Prefecture of Hunan Province., Methods: A total of 184 blood specimens were collected from Fenghuang, Huanyuan and Baojing counties of Xiangxi Autonomous Prefecture during the period from August 2018 through August 2019, and were detect using PCR assay with the specific 18S ribosomal rRNA ( 18S rRNA ) gene targeting Theileria . The gene sequences of positive specimens were aligned with the sequences recorded in GenBank, and a phylogenetic tree was created with Plasmodium ovale 18S rRNA as an outgroup., Results: A total of 143 blood samples were positive for Theileria , with a mean detection rate of 77.7%. Theileria was prevalent in the blood samples from yellow cattle in all three counties, with detection rates of 85.0% in Fenghuang County, 88.3% in Huayuan County and 61.0% in Baojing County, respectively. There was no significant difference in the detection rate of Theileria between Xiangxi yellow cattle and normal yellow cattle (77.2% vs. 79.5%; χ 2 = 0.08, P > 0.05), while the detection of Theileria was significantly lower in the housed yellow cattle than in free-range cattle (68.9% vs. 89.7%; χ 2 = 22.36, P < 0.01). A total of 18 PCR positive samples were randomly selected for sequencing and analysis, and all samples showed more than 99.0% homology with T. luwenshuni isolates. Phylogenetic analysis showed that the 18 positive samples were clustered into the same branch with T. luwenshuni , but were far away from other isolates., Conclusions: The prevalence of Theileria is high in yellow cattle from Xiangxi Autonomous Prefecture of Hunan Province, and T. luwenshuni may be the dominant parasite species.

Phthalates (PAEs) accumulated in agricultural soils and rice have increased human exposure risks. Microbial degradation could efficiently reduce the residue of organic pollutants in soil and crop plants. Here, we hypothesized that endophytic bacteria from wild rice have the potential for degradation of PAEs and plant growth promoting. The endophytic bacterial community and functional diversity in wild rice (Oryza meridionalis) were analyzed for the first time, and the potential for PAE degradation and plant growth promoting by endophytes were investigated. The results of Illumina high-throughput sequencing revealed that abundant endophytes inhabited in wild rice with Proteobacteria, Bacteroidetes, Firmicutes and Actinobacteria being the dominant phyla. Endophytic bacterial diversity and complexity were confirmed by isolation and clustering of isolates. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis demonstrated that endophytes exerted diverse functions such as plant growth promoting, xenobiotics biodegradation, pollution remediation and bacterial chemotaxis. Pure culture experiment showed that 30 isolated endophytic strains exhibited in vitro plant growth promoting activities, and rice plants inoculated with these strains confirmed their growth promoting abilities. Some endophytic strains were capable of efficiently degrading PAEs, with the highest removal percentage of di-n-butyl phthalate (DBP) up to 96.1% by Bacillus amyloliquefaciens strain L381 within 5 days. Synthetic community F and strain L381 rapidly removed DBP from soil (removing 91.0%-99.2% within 10 d and from rice plant slurry (removing 93.4%-99.2% within 5 d). These results confirmed the hypothesis and demonstrated the diversity of endophytic bacteria in wild rice with diverse functions, especially for plant growth promoting and removing PAEs. These multifunctional endophytic bacteria provided good alternatives to reduce PAE accumulation in crops and increase yield., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The mitochondrial DNA G-quadruplex (mtDNA G4) is a potential regulatory element for the regulation of mitochondrial functions; however, its relevance and specific roles in diseases remain largely unknown. Here, we engineered a set of chemical probes, including MitoISCH , an mtDNA G4-specific fluorescent probe, together with MitoPDS , a mitochondria-targeted G4-stabilizing agent, to thoroughly investigate mtDNA G4s. Using MitoISCH to monitor previously intractable dynamics of mtDNA G4s, we surprisingly found that their formation was prevalent only in endothelial and cancer cells that rely on glycolysis for energy production. Consistent with this, promotion of mtDNA G4 folding by MitoPDS in turn caused glycolysis-related gene activation and glycolysis enhancement. Remarkably, this close relationship among mtDNA G4s, glycolysis, and cancer cells further allowed MitoISCH to accumulate in tumors and label them in vivo. Our work reveals an unprecedented link between mtDNA G4s and cell glycolysis, suggesting that mtDNA G4s may be a novel cancer biomarker and therapeutic target deserving further exploration.

Because of the lack of facile and accurate methods to track stress granule (SG) dynamics in live cells and in vivo , in-depth studies of the biological roles of this attractive membraneless organelle have been limited. Herein, we report the first small-molecule probe, TASG , for the selective, convenient and real-time monitoring of SGs. This novel molecule can simultaneously bind to SG RNAs, the core SG protein G3BP1, and their complexes, triggering a significant enhancement in fluorescence intensity, making TASG broadly applicable to SG imaging under various stress conditions in fixed and live cells, ex vivo and in vivo . Using TASG , the complicated endogenous SG dynamics were revealed in both live cells and C. elegans . Collectively, our work provides an ideal probe that has thus far been absent in the field of SG investigations. We anticipate that this powerful tool may create exciting opportunities to investigate the underlying roles of SGs in different organisms.

Background: Severe acute pancreatitis (SAP) is a common acute abdominal disease with high morbidity and mortality. However, the mechanism underlying SAP is still unclear., Methods: Cerulean and LPS (Cer-LPS) was used to establish a rat model and an in vitro model of SAP. qRT-PCR, western blot and IHC were determined to analyse the expression of mRNA and proteins. IL-1β, TNF-α and IL-6 levels were measured applying ELISA. H&E staining was determined to observe the pathological changes. Apoptosis was tested by AV-PI staining using flow cytometry. CCK8 assay was taken to detect cell viability. Cell migration was assessed by transwell assay. Tube formation assay was conducted to evaluate angiogenesis. Luciferase assay was used to detect relationship of miR-20b-5p and AKT3., Results: MiR-20b-5p was lowly expressed in SAP models both in vivo and in vitro . Overexpression of miR-20b-5p restrained inflammation and apoptosis in Cer-LPS treated pancreatic acinar cells. Furthermore, miR-20b-5p promoted the angiogenesis of vascular endothelial cells, since the viability, migration and the capability of tube formation were increased by miR-20b-5p. Mechanically, miR-20b-5p directly targeted AKT3 to promote autophagy. Furthermore, miR-20b-5p could prevent the inflammation, apoptosis and enhance angiogenesis via enhancing autophagy, which was verified in vivo ., Conclusion: This study demonstrated miR-20b-5p attenuates SAP through directly targeting AKT3 to regulate autophagy, subsequently inhibit inflammation and apoptosis, and promote angiogenesis. Our findings suggested a novel target of miR-20b-5p for the therapy of SAP.

Bone remodeling is a process delicately balanced between osteoclastic bone resorption and osteoblastic bone formation. Osteoclasts (OCs) are multinucleated giant cells formed through the fusion of monocytic precursors of the hematopoietic stem cells lineage. OCs are the exclusive cells responsible for the resorption and degradation of the mineralized bone matrix. Pantoprazole (PPZ), a proton pump inhibitor (PPI), is commonly prescribed to reduce excess gastric acid production for conditions such as gastroesophageal reflux disease and peptic ulcer disease. Studies have found contradictory effects of PPI therapy on bone metabolism due to the lack of understanding of the exact underlying mechanism. In this study, we found that PPZ inhibits receptor activator of nuclear factor- κ B (NF- κ B) ligand- (RANKL-) induced osteoclastogenesis from bone marrow monocytic/macrophage (BMMs) precursors and the bone-resorbing activity of mature OCs. Correspondingly, the expression of OC marker genes was also attenuated. At the molecular level, PPZ treatment was associated with reduced activation of the ERK MAPK signaling pathways crucial to OC differentiation. Additionally, the in vivo administration of PPZ protected mice against lipopolysaccharide- (LPS-) induced inflammatory calvarial bone erosion, as a result of the reduced number and activity of OCs on the calvarial bone surface. Although PPI use is associated with increased risk of osteoporosis and bone fractures, our study provides evidence for the direct inhibitory effect of PPZ on OC formation and bone resorption in vitro and in vivo, suggesting a potential therapeutic use of PPZ in the treatment of osteolytic disease with localized bone destruction., Competing Interests: The authors have no conflicts of interest to declare., (Copyright © 2020 Yu-Xi Li et al.)

Aim: To investigate the diagnostic value of ultrasound elastography (UE) for benign and malignant axillary lymph nodes., Materials and Methods: A literature search was conducted from PubMed, Cochrane EMBASE, and Medline. Fourteen studies including 1,186 patients with 1,411 lymph nodes were enrolled. Overall, diagnostic descriptive statistics included pooled sensitivity, specificity, and area under the receiver operating characteristic curve (AUC) with corresponding 95% confidence intervals (95% CIs) were generated by random effect model. Subgroup analyses were performed in (real-time elastography [RTE] versus shear wave elastography [SWE]) and (conventional ultrasound versus combination of traditional ultrasound and elastography). Meta-regression was used to explore potential sources of heterogeneity., Results: The overall pooled sensitivity, specificity, and AUC of UE was 0.79 (95% CI: 0.71-0.86), 0.90 (95% CI: 0.83-0.95), and 0.91 (95% CI: 0.88-0.93), respectively. In the subgroup analysis of the two UE techniques, the pooled sensitivity, specificity, and AUC of SWE was higher than that of RTE (sensitivity: 0.82>0.77; specificity: 0.91>0.89; AUC: 0.94>0.89). The pooled diagnostic value of ultrasound combined with UE were significantly improving compared with traditional ultrasound (sensitivity: 0.87>0.82, specificity: 0.83>0.78, and AUC: 0.91>0.87). No independent heterogeneous factor was found in meta-regression., Conclusion: The results indicate that UE was an effective technique for identifying malignant axillary lymph nodes due to its high diagnostic efficiency, which can provide useful information for surgical procedure selection., (Copyright © 2020 The Royal College of Radiologists. Published by Elsevier Ltd. All rights reserved.)

Objective: To produce latent membrane protein 2A (LMP2A) chimeric antigen receptor (CAR)-T cells and detect the lethal effect of LMP2A CAR-T cells on nasopharyngeal carcinoma (NPC) cells. Methods: The study was conducted from September 2016 to December 2017.Genetic engineering technology was used to construct anti-LMP2A CAR lentiviral expression vector and sequencing was identified. The expression of anti-LMP2A CAR in the 293T cells was confirmed by western blot. CCK8 assay was used to evaluate the cytotoxicity of LMP2A CAR-T cells to NPC cells. ELISA assay was performed to test IL-2 and IFN-γ releasing of activated LMP2A CAR-T cells. The inhibition effect of LMP2A CAR-T cells on NPC xenograft tumor was observed in vivo . Statistical analysis was performed by statistical software SPSS 21.0. Results: The results of PCR and sequencing showed that anti-LMP2A CAR lentiviral expression vector was constructed successfully. The result of western blot indicated the expression of anti-LMP2A CAR in the 293T cells effectively. The results of CCK-8 assay showed that the killing activities of LMP2A CAR-T cells to LV-LMP2A-CNE1 cells were (72.11±9.75)%, (54.65 ±5.42)% and (36.68±3.80)% at 20∶1, 10∶1 and 5∶1 ratio of effective cells to target cells, and had a statistical difference compared to CD19 CAR-T cells and T cells ( P <0.05). There was no significant difference in the killing activities of LMP2A CAR-T cells to CNE1 cells compared with CD19 CAR-T cells and T cells. The results of ELISA showed that the content of IL-2 and IFN-γ in the co-culture supernatant of LMP2A CAR-T cells and LV-LMP2A-CNE1 cells was significantly higher than that of LMP2A CAR-T cells and CNE1 cells which had statistical difference ( P <0.05); In vivo experiment, the volume of LMP2A CAR-T cell group was (80.3±10.0) mm(3) which was significantly lower than that of the control groups, and the difference was statistically significant ( P <0.05). Conclusion: LMP2A CAR-T cells are successfully prepared and have an obvious targeting cytotoxicity on LMP2A-positive NPC cells.

DEP domain containing 1 (DEPDC1) is recently reported to be overexpressed in several types of human cancer; however the role of DEPDC1 in prostate cancer remains to be investigated. Herein, we identified that the DEPDC1 mRNA and protein expression levels were dramatically increased in prostate cancer tissues and cell lines. Overexpression of DEPDC1 promoted, but depletion of DEPDC1 inhibited cell proliferation by regulating the G1-S phase cell cycle transition. Importantly, we found that DEPDC1 was essential for the tumor growth and formation of bone metastases of prostate cancer cells in vivo. Finally, we demonstrated that DEPDC1 interacted with E2F1 and increased its transcriptional activity, leading to hyper-activation of E2F signaling in prostate cancer cells. Our findings reveal an oncogenic role of DEPDC1 in prostate cancer progression via activation of E2F signaling, and suggest DEPDC1 might be a potential therapeutic target against the disease., (Copyright © 2017 Elsevier Inc. All rights reserved.)

The frequent modification of cell-surface proteins by N-linked glycans is known to be correlated with many biological processes. Aberrant glycosylation on surface proteins is associated with different cellular statuses and disease progression. However, it is extraordinarily challenging to comprehensively and site-specifically analyze glycoproteins located only on the cell surface. Currently mass spectrometry (MS)-based proteomics provides the possibility to analyze the N-glycoproteome, but effective separation and enrichment methods are required for the analysis of surface glycoproteins prior to MS measurement. The introduction of bio-orthogonal groups into proteins accelerates research in the robust visualization, identification, and quantification of proteins. Here we have comprehensively evaluated different sugar analogs in the analysis of cell-surface N-glycoproteins by combining copper-free click chemistry and MS-based proteomics. Comparison of three sugar analogs, N-azidoacetylgalactosamine (GalNAz), N-azidoacetylglucosamine (GlcNAz), and N-azidoacetylmannosamine (ManNAz), showed that metabolic labeling with GalNAz resulted in the greatest number of glycoproteins and glycosylation sites in biological duplicate experiments. GalNAz was then employed for the quantification experiment in statin-treated HepG2 liver cells, and 280 unique N-glycosylated sites were quantified from 168 surface proteins. The quantification results demonstrated that many glycosylation sites on surface proteins were down-regulated in statin-treated cells compared to untreated cells because statin prevents the synthesis of dolichol, which is essential for the formation of dolichol-linked precursor oligosaccharides. Several glycosylation sites in proteins that participate in the Alzheimer's disease pathway were down-regulated. This method can be extensively applied for the global analysis of the cell-surface N-glycoproteome.

Objective: Gynecological cancer survivors are at increased risk of psychological problems including suicide risk. Suicidal ideation, which was thought to be precursor to suicide attempts, has not been well studied. This study aimed to investigate the prevalence, and determinants of suicidal ideation for women with gynecological cancer, and then to assess the effect of coping style and social support on suicidal ideation., Methods: Patients with cervical, ovarian and endometrial cancers seen at Hunan Provincial Tumor Hospital from September 2012 to June 2013 were consecutively recruited and were asked to complete the Zung Self-Rating Depression Scale, Suicidal Ideation of Self-rating Scale, Medical Coping Modes Questionnaire and Social Support Rating Scale. Path analysis was used to examine the relationship among coping style, social support, depression symptoms and suicidal ideation., Results: A total of 579 (579/623, 93.0%) gynecological cancer patients were enrolled in this study and completed all investigations between September 2012 and June 2013. Among them, 105 (18.1%) patients reported suicidal ideation, with the highest rate in patients with ovarian cancer (30.16%). Suicidal ideation was associated with depression symptoms, care providers, chemotherapy history and acceptance-resignation. Path analysis showed that the acceptance-resignation affected suicidal ideation directly as well as mediated by social support and depression symptoms, while confrontation and avoidance affected suicidal ideation entirely through social support and depression symptoms., Conclusions: Suicidal ideation is high among patients with gynecological cancer, especially among ovarian cancer patients. Coping strategies such as confrontation and avoidance, and social support may be helpful for preventing suicidal ideation among them., (Copyright © 2015 John Wiley & Sons, Ltd.)

Background: Preoperative conditions may play a significant role in postoperative cognitive dysfunction (POCD) development in elderly patients. We aimed to investigate whether preoperative cognitive training could lower the incidence of POCD one week after surgery., Material and Methods: A total of 141 ASA I-III elderly patients who underwent gastrointestinal surgery were enrolled into the study. Patients were randomized into either the Intervention group (69 analyzed) or the Control group (72 analyzed). Patients in the intervention group were instructed and trained in a cognition mnemonic skill for a total of three 1-hour sessions with the method of loci (MoL). Controls did not receive any cognitive training during hospitalization. All patients were tested using neuropsychological battery tests (NPTs) on admission and one week after surgery., Results: The incidence of POCD in the intervention group (15.9%) was significantly lower than in the controls (36.1%) (P<0.05). Patients' performance in Brief Visuospatial Memory Test-Revised and Symbol-Digit Modalities Test were improved by the cognitive training. Increasing age, longer length of anesthesia and surgery, and lack of cognitive training were associated with a significantly higher risk of POCD (P<0.05)., Conclusions: Cognitive training with MoL can reduce the decline of early postoperative cognitive function in elderly patients undergoing major gastrointestinal surgery.

For decades, statins have been widely used to lower cholesterol levels by inhibiting the enzyme HMG Co-A reductase (HMGCR). It is well-known that statins have pleiotropic effects including improving endothelial function and inhibiting vascular inflammation and oxidation. However, the cellular responses to statins and corresponding pleiotropic effects are largely unknown at the proteome level. Emerging mass spectrometry-based proteomics provides a unique opportunity to systemically investigate protein and phosphoprotein abundance changes as a result of statin treatment. Many lipid-related protein abundances were increased in HepG2 cells treated by atorvastatin, including HMGCR, FDFT, SQLE, and LDLR, while the abundances of proteins involved in cellular response to stress and apoptosis were decreased. Comprehensive analysis of protein phosphorylation demonstrated that several basic motifs were enriched among down-regulated phosphorylation sites, which indicates that kinases with preference for these motifs, such as protein kinase A and protein kinase C, have attenuated activities. Phosphopeptides on a group of G-protein modulators were up-regulated, which strongly suggests that cell signal rewiring was a result of the effect of protein lipidation by the statin. This work provides a global view of liver cell responses to atorvastatin at the proteome and phosphoproteome levels, which provides insight into the pleiotropic effects of statins.

Objective: To investigate the current distribution of Paragonimus westermani in Guangdong Province., Method: Snails and crabs collected from mountain streams in regional survey sites were dissected to detect cercarial and metacercarial infections of P. westermani. Domestic cats and dogs artificially infected with the collected metacercariae were also dissected to detect adult worms of P. westermani. The COI and ITS2 gene sequences of those adult worms were compared with those of known Paragonimus specimen deposited in the GenBank., Results: All of the first intermediate hosts in five survey sites of Liangkou, Nankun, Mountain, Dadong, Muxi, Guowu, were identified as Semisulcospira libertina, whose cercariae infection rates were 0.33%, 0.15%, 0.058%, 0.10%, and 0.05%, respectively; the second intermediate hosts in above five sites were all identified as Sinopotamon denticulatum, whose metacercariae infection rates were 100%, 100%, 38.09%, 55.36%, and 65.26%, respectively. The numbers of metacercariae in the five sites were 79.4, 105.66, 9.16, 16.18, and 15.6 per positive crab, respectively, and 11.12, 7.87, 0.58, 0.69, and 0.85 per gram of crab, respectively. All the metacercariae were identical to those of P. westermani. Adult worms and eggs of P. westermani were found in both reservoir hosts of domestic cats and dogs infected artificially. By comparing the COI genes of five representative samples from each survey site with that of Paragonimus #AF219379.21, AF540958.1 from GenBank, we found out the homology to be 99%, 99%, 99%, 98%, and 99%, respectively. In addition, a comparison of the ITS2 gene sequences between the above five samples and Paragonimus #DQ836243.1, DQ351845.1, AB354217.1 from GenBank revealed 98%, 99%, 98%, 98%, and 98% gene homology, respectively., Conclusion: Two ultra-high and three high endemic areas of P. westermani are discovered in Guangdong Province. No obvious differences were found among the types of P. westermani in the above five endemic areas.

The Agrobacterium-mediated transformation system is the most commonly used method in soybean transformation. Screening of soybean genotypes favorable for Agrobacterium-infection and tissue regeneration is the most important step to establish an efficient genetic transformation system. In this study, twenty soybean genotypes that originated from different soybean production regions in China were screened for transient infection, regeneration capacity, and stable transgenic efficiency. Three genotypes, Yuechun 04-5, Yuechun 03-3, and Tianlong 1, showed comparable stable transgenic efficiencies with that of the previously reported American genotypes Williams 82 and Jack in our experimental system. For the Tianlong 1, the average stable transformation efficiency is 4.59%, higher than that of control genotypes (Jack and Williams 82), which is enough for further genomic research and genetic engineering. While polymerase chain reaction (PCR), LibertyLink strips, and β-glucuronidase (GUS) staining assays were used to detect the insertion and expression of the transgene, leaves painted with 135 mg/L Basta could efficiently identify the transformants.

Innate immune response plays the key role in initiating and guiding the immune response. Elucidating the innate immune related molecular events involved in the interaction between the parasite and the host will aid in the development of an effective vaccine and anti-schistosome pharmaceuticals. In this study, we examined the regulatory effect of Schistosoma japonicum soluble egg antigen (SEA) on MHC class II expression in macrophage cell line RAW 264.7. We demonstrated that SEA possesses the ability to down-regulate IFN-γ-induced MHC class II expression in RAW 264.7 cells. The production of IL-10 and IL-6 in RAW 264.7 cells, induced by SEA, was responsible for mediating the down-regulation of MHC class II. Our findings suggest that in RAW 264.7 cells (1) IFN-γ provides a condition for lower concentrations of SEA to attenuate MHC class II expression; (2) SEA attenuated IFN-γ-induced MHC class II expression and the IL-10 and IL-6 production is mediated at least partly by the interaction of SEA with TLR4; and (3) SEA attenuated IFN-γ-induced MHC class II expression at the transcriptional level.

Objective: To investigate the ultrastructure and the alkaline phosphatase (ALP) activity changes of NIH3T3 cells incubated with secretive human bone morphogenetic protein-2 (hBMP-2) that is induced by gene transfection through transwell system., Methods: Eukaryotic expression vector (pcDNA3.1-B2) was transduced into NIH3T3 cells by Sofast, a positive compound transfection agent. The positive cell clones were selected with G418. The cytoplasmic and extracellular expression of BMP-2 in the NIH3T3 cells were determined by immunohistochemical and enzyme-linked immunosorbent assay (ELISA). NIH3T3 cells were co-cultured with hBMP-2 gene transfecting cells through transwell system, and the ultrastructure and ALP activity (the markers of osteogenetic differentiation) changes were observed., Results: There were cytoplasmic and extracellular expression of BMP-2 in transfecting NIH3T3 cells. The ultrastructure changes and the high expression of ALP suggested the osteogenetic differentiation tendency of NIH3T3 cells co-cultured with transfecting NIH3T3 cells., Conclusions: Secretive BMP-2 that is induced by gene transfection could promote the osteogenetic differentiation of fibroblast cells.

Background: Bone morphogenetic proteins (BMPs), which belong to the transforming growth factor beta superfamily, are powerful regulators of cartilage and bone formation. This study investigated the biological changes of NIH3T3 cells incubated with secretive BMP2 that was induced by gene transfection through transwell., Methods: Eukaryonic expression vector (pcDNA3.1-B2) was transfered into NIH3T3 cells with Sofast, a positive compound transfection agent. The positive cell clones were selected with G418. The cytoplasmic and extracellular expressions of BMP2 were determined by immunohistochemical stain and enzyme-linked immunosorbent assay. NIH3T3 cells were co-cultured with hBMP2 gene transfecting cells through transwell, and the ultrastructure, alkaline phosphatase activity and the expression of osteocalcin (the marker of osteogenetic differentiation) changes were observed., Results: There were cytoplasmic and extracellular expressions of BMP2 in transfecting NIH3T3 cells. The ultrastructural changes, the high activity of alkaline phosphatase and the positive stain of osteocalcin suggested the osteogenetic differentiation tendency of NIH3T3 cells co-cultured with transfecting NIH3T3 cells., Conclusion: Secretive BMP2 that is induced by gene transfection could promote the osteogenetic differentiation of fibroblast cells.

Objective: To determine optimum culture conditions for the seed embryo culture and rapid propagation of Dendrobium candidum., Method: Seed embryos of D. candidum were incubated in the medium containing a combination of 6-benzylaminopurine (BA) and 1-naphthaleneacetic acid (NAA), potato extract, banana extract and activated carbon in order to induce seed embryo germination, protocorm differentiation, plantlet propagation and plantlet rooting., Result and Conclusion: The maximum embryo germination percentage was obtained in the 1/2 MS media supplemented with 20% potato extract. The 1/2 MS medium supplemented with 1.0 mg x L(-1) BA and 0.1 mg x L(-1) NAA was very beneficial to the protocorm differentiation and propagation of D. candidum. The highest protocorm propagation index was obtained from the medium containing the activated carbon. The highest root numbers and length were observed in plants growing in 1/2 MS medium containing 0.5 mg x L(-1) NAA.

Objective: To study the feasibility of diagnosing parotid disease with magnetic resonance sialography (MRS) and to select the optimal scanning sequence., Methods: Twenty-three patients with parotid gland disease and 5 normal adults received sialography using magnetic resonance imaging technique and several sequences (including IR-FSE, FSE, SS-IR-FSE, SS-FSE) were used. After first scanning, the patients were scanned respectively 3 and 10 minutes after buccal application of vitamin C. And MR images of duct obtained. The images of parotid duct system were analysed and evaluated according to their displaying effects. Qualitative diagnosis was made based on MRI and those diagnosis were compared with pathological diagnosis after operation., Results: Images of MR sialography clearly displayed the main duct and its branches of parotid gland and the pathological changes of duct, such as narrow, expanded, stoppage. Of the scanning sequences, IR-FSE was superior to others in manifesting the parotid gland duct (P < 0.05). The performance of images after being given vitamin C did not significantly improve the displaying effect. The accurate rate of qualitative diagnosis was 95.6%., Conclusions: MR sialography can clearly display the parotid ducts and their pathological changes. The accurate rate of qualitative diagnosis of parotid disease was higher than that X-ray sialography.

Objective: To observe the structure-activity and time-effect relations of the central stimulating effects of modafinil for further pharmaceutical design and development and rational use of this kind of stimulant., Method: Male mice served as subjects. The experiment was divided into two parts: 1) comparison between the dose (60, 120, 240 mg/kg) and temporal features of the effects of modafinil and its two derivatives on locomotor activity of mice; 2) Observation of the stimulating effects of modafinil (120 mg/kg) given during the day or night., Result: Modafinil could significantly increase the locomotor activity of mice in a dose-dependent way, the effect was best observed at the concentration of 120 mg/kg. The stimulating action of modafinil was increased by chlorination, but decreased by methylation. The central stimulating effect of modafinil showed no significant circadian difference., Conclusion: The central stimulating effect of modafinil shows dose and time-related features. The effect tends to be enhanced by Chlorination but reduced by methylation.

Objective: To assess the risk factors for vulvar dystrophy., Methods: An epidemiological study was carried out. Data on 100 cases with vulvar dystrophy was reviewed and face to face interviewed with a uniform questionnaire including the manner of work, environmental temperature, habit of eating, mood, underwear wearing, autoimmune diseases, marriage, menstrual age, the quantity of menses, orders of pregnancy, and labor trauma of vulvar during delivery, vulvitis and urethritis ect. Univariable analysis and multivariate logistic regression analysis were carried out with 1:1 case-control methodology., Results: Multiple conditional logistic regression analysis showed that vulvar dystrophy was positively associated with hot food (OR = 2.55, 95% CI: 1.24 - 5.25), mood (OR = 4.27, 95% CI: 1.96 - 9.29), order of pregnancy (OR = 3.37, 95% CI: 2.11 - 5.40), vulvitis (OR = 6.74, 95% CI: 2.66 - 17.09) and urethritis (OR = 11.02, 95% CI: 1.01 - 120.19). Vulviitis or urethritis increased 6.74 or 11.02 times the incidence of vulva dystrophy. Anger or nervous state contributed to the incidence of vulva dystrophy (OR = 4.27). Addict to hot food and order of labor also increased risk ratio for 2.55 and 3.37 times, respectively., Conclusion: The risk factors of vulvar dystrophy were: addict to hot food, often holding a angry or nervous state, increase of labors, having vulvitis and urethritis.

Objective: To investigate the effects of modafinil, a new central stimulant, on visual and auditory motion reaction ability and subjective fatigue level during 48 h sleep deprivation (SD)., Method: Six male healthy young volunteers were exposed to two 48 h periods of continuous wakefulness during the crossover experiment. In one period, three 200 mg doses of modafinil were given and in the other, separated by two weeks, matching placebos were administered. The SD time started from 7:00 of the first day to 7:00 of the third day. Drugs were given at 0:00, 16:00 of the second day and 0:00 of the third day. Variables, including visual and auditory motion reaction time, attention distribution, critical flicker fusion frequency (CFF), Stanford sleepiness scale (SSS) and rating of perceived exertion (RPE), were tested at 21:00 of the first day and 1, 3, 5, 7 h after each drug administration., Result: After 2-3 does of modafinil, the CFF level was obviously enhanced while the scores of Stanford sleepiness scale and RPE were markedly reduced. The visual and auditory motion reaction ability, as well as the attention distribution, showed no significant change., Conclusion: Modafinil can effectively reduced the subjective fatigue and sleepiness levels during SD.

Objective: To investigate the change of lipid peroxidation in various organs of rats after repeated +10 Gz stress., Method: Twenty male Wistar rats were randomly divided into control (+1 Gz) group and +10 Gz group (n = 10 for each group ). The two groups were exposed to +1 Gz or +10 Gz on an animal centrifuge respectively. The onset rate of +Gz was 0.5 G/s; the sustained time at peak +Gz was 30 s each time, with 5 times/d, 1 min intervals, 3 d/wk, 3 wk in total. The rats were decapitated in ice bath on the next day after the last +Gz exposure. The lung, heart, liver and kidney of rats were collected for the following determinations. These organs were made into homogenates and the mitochondria of heart and kidney were isolated from their homogenates. Then malondialdehyde (MDA) content , superoxide dismutase (SOD) activity and glutathion (GSH) concentration in these homogenates and mitochondria were determined individually., Result: Compared with the control, MDA concentration in myocardial mitochondria increased significantly (P < 0.01) after repeated +10 Gz exposures, and the SOD activity in liver homogenate and renal mitochondria decreased remarkably (P < 0. 01). But there was no significant difference between GSH concentrations of the two groups., Conclusion: Repeated +10 Gz stress induced lipid peroxidation injury of myocardial mitochondria and had some influences on the oxygen radical metabolism of liver and kidney in rats.

Objective. To observe the effects of tea polyphenols (TP) on cardiac function and myocardial ultrastructure in rats after repeated +10 Gz stress. Method. Twenty four male Wistar rats were randomly divided into three groups (n=8 each): group A (control), group B (+10 Gz), group C (+Gz with TP). Group B and C were repeatedly exposed to +10 Gz (each for 30 s, onset rate about 0.5 G/s, 3 times/d with +1 Gz 1 min intervals, 3 d/wk, 4 weeks in total), but group A was only submitted to +1 Gz. TP (200 mg kg-1) was given orally to group C about 1 h prior to the +Gz experiment, and distilled water was given to group A and B. Function of isolated rat working hearts and myocardial ultrastructure were observed. Result. A significant decrease of left ventricular systolic pressure (LVSP) and injury of myocardial structure in rats were demonstrated after repeated +10 Gz stress. But TP could remarkably elevate the LVSP and improve myocardial ultrastructural injury in +10 Gz stressed rats. Conclusion. These results indicated that repeated high G exposure may produce cardiac structural and functional injuries in rats which might be partly related to free radical metabolism; and antioxidant TP had significant protective effects on the hearts of +Gz stressed rats.

Intracellular potentials were recorded from nociceptive neurons(NCNs) in primary somatosensory cortex area (SI) of cats in response to stimulation of ventral posterior lateral nucleus (VPL). When the polarizing current injected into NCNs was < or = 1.0 nA, the I-V curve was quite linear (r=0.96). However, when the polarizing current was >1.0 nA, rectification rendered the I-V curve sigmoid in shape. The rectification in NCNs was more prominent than that in NNCNs. Rm,tau and Cm of the NCNs were obviously larger than those of NNCNs (P<0.01 or P<0.05). The responses of NCNs evoked by stimulating VPL and SN might be similar or different. Implications of the differences of NCN and NNCN were discussed.

Objective: To observe the effects of repeated +10 Gz stress on cerebral lipid peroxidation, liver and renal function in rats and the prophylactic effects of antioxidant tea polyphenols (TP)., Method: Twenty-four male Wistar rats were randomly divided into three groups (n = 8, each): group A (control), group B (+10 Gz), and group C (TP). Group B and C were exposed to repeated +10 Gz stress (each for 30s, onset rate about 0.5 G/s, 3 times/d with +1 Gz 1 min intervals, 3 d/wk, 4 weeks in total), but group A was only submitted to +1 Gz. TP (200 mg/kg) was given orally in group C about 1 h prior to the +Gz experiment, while distilled water was given in group A and B. Lipid peroxidation in the brain, liver and renal functions and serum lipids were determined., Results: As compared with the control, lipid peroxidation in rat cerebral homogenate, mitochondria and cytoplasm was significantly increased (P < 0. 05), and serum creatinine concentration was markedly elevated after repeated +10 Gz stress (P < 0. 01). But, TP had significant inhibitory effect on +10 Gz stress induced peroxidative injury in rat brain and reduced the serum creatinine level. There were no differences of serum triglyceride and total cholesterol concentrations and glutamic-pyruvic transaminase activity among the three groups., Conclusion: These results indicated that repeated high +Gz stress could bring about peroxidative injury in brain and harmful effect on renal function, and natural antioxidant TP had significant protective effects.

Monosaccharide composition was determined in apolipoprotein B-48 (apoB) of chylomicrons of rat mesenteric lymph. Chylomicrons were separated into three fractions based on density. Triglyceride and apolipoprotein content were determined in each. ApoB was isolated and quantified using precipitation with isopropanol. Chylomicrons were collected in lymph under normal conditions, and with Poloxalene 2930 when chylomicron secretion was inhibited. Most of the triglyceride was carried in the least dense fraction, while the highest apoB content was in the most dense fraction under normal conditions. Mannose and galactosamine contents of apoB were similar in all fractions while contents of both glucosamine and galactose were highest in the least dense fraction. When chylomicron secretion was inhibited by Poloxalene, the amount of triglyceride recovered in the least dense fraction was significantly reduced. Despite the inhibition of lipid transport in the least dense fraction of chylomicrons by Poloxalene, there was little change in apoB recoveries and in the relative content of various monosaccharides in the apoB from each of the three fractions as compared to results obtained during lipid absorption under normal conditions. In conclusion, carbohydrate composition of apoB of chylomicrons is heterogeneous and varies with chylomicron density.

Postoperative anastomotic leakage is one of the most dangerous and, unfortunately, most frequent complications. Many methods have been tried to prevent such leakage. The incidence of leakage has decreased to 3.84% in our hospital in recent years. Ten cases of gastroesophageal anastomotic leakage following resection of carcinoma of the esophagus and gastric cardia are analyzed and a new tool, the three-leaf clamp, which reduced the anastomotic leakage, is introduced.