A bioinformatics analysis of the susceptibility genes in Stanford type A aortic dissection.

Background: The expression levels of long noncoding RNAs (lncRNAs) and mRNAs in human acute Stanford type A aortic dissecting aneurysm and normal active vascular tissues were compared using the array lncRNA/mRNA expression profile chip technology. Methods: The tissue samples of 5 patients who presented with Stanford type A aortic dissections and the normal ascending aorta tissues from 5 donor heart transplantation patients receiving surgical treatment in Ganzhou People's Hospital were collected. Hematoxylin and eosin (HE) staining were performed to investigate the structural features of the ascending aortic vascular tissue. Nanodropnd-100 was used to detect the surface level of RNA in 10 samples included in the experiment, to ensure that the quality of the standard was consistent with the core plate detection. NanoDrop ND-1000 was used to detect the RNA expression levels in 10 specimens included in the experiment to ensure that the quality of specimens satisfied the requirements of the microarray detection experiment. The Arraystar Human LncRNA/mRNAV3.0 expression profile chip (8×60K, Arraystar) was used to detect the expression levels of lncRNAs and mRNAs in the tissue samples. Results: A total of 29,198 lncRNAs and 22,959 mRNA target genes could be detected in the above tissue samples after the initial data were standardized and low-expression information was filtered. The data in the middle of the range of 50% value consistency was higher. The scatterplot results preliminarily suggested that there were large numbers of lncRNAs with increased and decreased expression in Stanford type A aortic dissection tissues compared with normal aortic tissues. The differentially expressed lncRNAs were enriched in BPs including apoptosis, nitric oxide synthesis, estradiol response, angiogenesis, inflammatory response, oxidative stress, and acute response; cell components (CCs) including cytoplasm, nucleus, cytoplasmic matrix, extracellular space, protein complex, and platelet α granule lumen; and MFs including protease binding, zinc ion binding, steroid compound binding, steroid hormone receptor activity, heme binding, protein kinase, cytokine, superoxide dismutase, and nitric oxide synthase activities. Conclusions: Gene ontology analysis demonstrated that many genes in Stanford type A aortic dissection were involved in cell biological functions, cell components, and molecular functions through upregulating and downregulating the levels of expression.

A study of clinical implementation of ventricular septal defect closure using three transthoracic approaches.

OBJECTIVE: To investigate the clinical implementation of ventricular septal defect closure using the three transthoracic approaches. METHODS: A total of 70 children with septal defects admitted to our hospital from January 2017 to December 2020 were selected as the study subjects. Among them, 10 children with the left thorax-right ventricle-left ventricle approach were assigned to Group A, 8 children with the right thorax-atrium dextrum-right ventricle-left ventricle approach were assigned to Group B, and 52 children with the subxyphoid-right ventricle-left ventricle approach were assigned to Group C. The surgical indices were recorded, the success rates of closure and cardiopulmonary function indices were compared, electrocardiogram (ECG), echocardiogram and X-ray film were investigated at 1, 3 and 12 months after surgery, and the incidence of complications was recorded. RESULTS: There was no statistically significant difference in the success rate of closure among the three groups (P > 0.05). The duration of intracardiac operations in Groups A and C was remarkably shorter than that in Group B, and the duration of skin incision and suture and hospital stay in Groups A and B were noticeably shorter than those in Group C (P < 0.05). After surgery, there was statistically significant difference in the contents of creatine kinase MB (CK-MB) isoenzyme, lactate dehydrogenase (LDH), serum malondialdehyde (MDA) and superoxide dismutase (SOD) among the three groups (P > 0.05). Airway resistance (Raw), oxygenation index (OI), and alveolar-arterial oxygen gradient (AaDO2) indicated that the postoperative pulmonary function in Group C was more effectively protected. There was no statistically significant difference in the incidence of complications among the three groups (P > 0.05). CONCLUSION: Ventricular septal defect closure using the three transthoracic approaches exhibited a high success rate and a high safety profile.

Analysis of poor prognostic factors of cerebral nerve after deep hypothermic circulatory arrest in patients with type A aortic dissection.

OBJECTIVE: To investigate the influences of deep hypothermic circulatory arrest (DHCA) on postoperative cranial nerve function in patients undergoing surgery for type A aortic dissection. METHODS: A total of 100 patients undergoing DHCA during the surgery for type A aortic dissection in our hospital were selected as the study subjects. After surgery, 32 patients with neurological complications were assigned to Group A, and 68 patients without neurological complications were assigned to Group B. The clinical outcomes were compared between the two groups, and the risk factors of postoperative neurological complications were analyzed by univariate and multivariate logistic regression analysis. RESULTS: During the surgery, patients underwent cerebral perfusion at 5 min and 10 min during DHCA had remarkably decreased cerebral oxygen saturation (rSO2) and VmMCA than those before anesthesia induction (P<0.05). After recovery of CPB, rSO2 and mean velocity in middle cerebral artery (VmMCA) recovered to the preoperative levels. The correlation analysis revealed a positive correlation between rSO2 and VmMCA (P<0.05). The univariate analysis suggested that the history of hypertension, hydropericardium, surgical duration, duration of cardiopulmonary bypass (CPB), aortic occlusion, ICU, and ventilator-assisted respiration, and hypoxemia significantly affected postoperative cranial nerve function (P<0.05). The logistic multivariate regression analysis demonstrated that the duration of CPB and aortic occlusion and hypoxemia were independent risk factors for postoperative cranial nerve dysfunction (P<0.05). CONCLUSION: There were noticeable changes in hemodynamic and blood oxygen parameters in patients with type A aortic dissection undergoing DHCA during the perioperative period. The long duration of CPB and aortic occlusion and preoperative hypoxemia are the independent risk factors leading to postoperative impaired cranial nerve function.

Intelligent photothermal dendritic cells restart the cancer immunity cycle through enhanced immunogenic cell death.

Dendritic cells (DCs) play a pivotal role in initiating antigen-specific tumor immunity. However, the abnormal function of DCs owing to the immunosuppressive tumor microenvironment (TME) and the insufficient number of tumor infiltrating DCs could promote immune tolerance and tumor immune escape. Thus, there is great potential to employ DCs to induce efficient antitumor immunity. In this paper, we developed intelligent DCs (iDCs), which consist of nanoparticles loaded with photothermal agents (IR-797) and coated with a mature DC membrane. The DC cell membrane on the surface of iDCs preserves the ability to present antigens and prime T cells. The iDCs can also enter the lymph node and stimulate T cells. The activated T cells reduced the expression of heat shock proteins (HSPs) in tumor cells, rendering them more sensitive to heat stress. Subsequently, we used mild photothermal therapy (42-45 °C) to induce immunogenic cell death and contribute to a synergistic antitumor effect. iDCs as a refined and precise system in combination with DC-based immunotherapy and thermal therapy can be stored long-term and on a large scale, so they can be applied in many patients.

Natural-Killer-Cell-Inspired Nanorobots with Aggregation-Induced Emission Characteristics for Near-Infrared-II Fluorescence-Guided Glioma Theranostics.

Nature has always inspired robotic designs and concepts. It is conceivable that biomimic nanorobots will soon play a prominent role in medicine. The "Terminator" in the science fiction film is a cybernetic organism with living tissue over a metal endoskeleton, which inspired us to develop natural-killer-cell-mimic nanorobots with aggregation-induced emission (AIE) characteristics (NK@AIEdots) by coating a natural kill cell membrane on an AIE-active polymeric endoskeleton, PBPTV, a highly bright NIR-II AIE-active conjugated polymer. Owing to the AIE and soft-matter characteristics of PBPTV, as-prepared NK@AIEdots maintained a superior NIR-II brightness (quantum yield ∼7.9% in water) and good biocompatibility. Besides, they can serve as a tight junction (TJ) modulator to trigger an intracellular signaling cascade, causing TJ disruption and actin cytoskeleton reorganization to form an intercellular "green channel" to help them to cross the blood-brain barrier (BBB) silently. Furthermore, they can initiatively accumulate in glioblastoma cells in the complex brain matrix for high-contrast and through-skull tumor imaging. The tumor growth was also greatly inhibited by these NK@AIEdots under the NIR light illumination. As far as we know, the quantum yield of PBPTV is the highest among the existing NIR-II luminescent conjugated polymers. Besides, the NK-cell biomimetic nanorobots showed great potential for BBB-crossing active delivery.

PPAR γ/Nnat/NF-κB Axis Involved in Promoting Effects of Adiponectin on Preadipocyte Differentiation.

A previous study has demonstrated that adiponectin (APN) could promote preadipocyte differentiation, and the present study further explored its mechanism. 3T3-L1 cells were infected with adenovirus holding human adiponectin gene apM1 and mouse neuronatin (Nnat) shRNA and initiated differentiation while coculturing with mature adipocytes stimulated with LPS. After 8 days, preadipocyte differentiation was observed by Oil Red O staining. Real-time quantitative PCR was used to evaluate mRNA expression levels of monocyte chemoattractant protein-1 (MCP-1), interleukin- (IL-) 6, IL-8, and tumor necrosis factor α (TNF-α). The levels of reactive oxygen species (ROS), total antioxidant capacity (T-AOC), malondialdehyde (MDA), and superoxide dismutase (SOD) in 3T3-L1 cells were detected. Western blotting was done to quantify the protein expression levels of Nnat, peroxisome proliferator-activated receptor (PPAR) γ, p65, and inhibitor of nuclear factor κB (IκB) α. Results demonstrated that APN overexpression markedly increased preadipocyte differentiation; inhibited gene expression of MCP-1, IL-6, IL-8, and TNF-α; reduced ROS and MDA release; increased T-AOC and SOD levels; upregulated Nnat, PPAR γ, and IκB α protein expressions; and downregulated p65 protein expression under LPS stimulation. However, the effects of APN were markedly attenuated when Nnat expression was knocked down. Taken together, the present study provided evidences that the effects of APN on promoting preadipocyte differentiation under inflammatory conditions via anti-inflammation and antioxidative stress may be regulated by the PPAR γ/Nnat/NF-κB signaling pathway.

Scaffolds biomimicking macrophages for a glioblastoma NIR-Ib imaging guided photothermal therapeutic strategy by crossing Blood-Brain Barrier.

Glioblastoma (GBM) is one of the most malignant cancers, and Blood-Brain Barrier (BBB) is the main obstacle to diagnose and treat GBM, hence scientists are making great efforts to develop new drugs which can pass BBB and integrate diagnosis and therapeutics together. Here, we designed plasma membrane of macrophage camouflaged DSPE-PEG loaded near-infrared Ib (NIR-Ib) fluorescence dye IR-792 nanoparticles (MDINPs). MDINPs were able to penetrate BBB and selectively accumulate at tumor site, and then could be used as NIR-Ib fluorescence probes for targeted tumor imaging. At the same time, MDINPs could kill tumor cells by photothermal effect. Our results showed that MDINPs-mediated NIR-Ib fluorescence imaging could clearly observe orthotopic GBM, and the NIR-Ib imaging-guided photothermal therapy significantly suppressed the growth of GBM and prolonged the life of mice. This work not only provided a method to mimic the biological function of macrophage, but also provided an integrative strategy for diagnosis and treatment in GBM.

Enzyme and pH dual-responsive hyaluronic acid nanoparticles mediated combination of photodynamic therapy and chemotherapy.

Hyaluronic acid (HA) is a natural biopolymer that can target to tumor cells due to CD44 receptors overexpressed in tumor cells. Here, a theranostic nanoparticle HA-Ce6 (DOX) with enzyme and pH dual-responsive is presented, which combined HA and a highly promised photosensitizer chlorin e6 (Ce6) using adipic dihydrazide (ADH) as a linker. The hydrazide group on its surface can efficiently conjugate doxorubicin to form HA-Ce6 (DOX) nanoparticles through the pH-sensitive hydrazone bond. In this study, the dual-response of HA-Ce6 (DOX) nanoparticles in the tumor cell are discussed. The HA-Ce6 (DOX) nanoparticles showed an average size of 90 nm with a uniform spherical morphology. In vitro drug release studies showed that HA-Ce6 (DOX) accomplished rapid drug release under acid conditions and enzyme stimulating. Confocal images revealed that the nanoparticles enhance the cellular accumulation of DOX and Ce6 in A549 cells. The therapeutic efficacy of HA-Ce6 (DOX) nanoparticles in A549 cells in vitro was evaluated through the MTT assay. The results showed that the therapeutic efficacy of HA-Ce6 (DOX) nanoparticles against A549 cells was remarkably enhanced compared with free DOX and free Ce6. These results indicate that the HA-Ce6 (DOX) nanoparticles could be a promising delivery system for photodynamic therapy and chemotherapy.

Near-infrared fluorescence imaging for vascular visualization and fungal detection in plants.

We found that heptamethine dye IR-820 showed distinct emission peaks in both the NIR-Ia and NIR-Ib windows. IR-820 yielded images of vascular structures in the NIR-Ib window with unprecedented details. NIR-Ib fluorescence imaging was useful not only for studying plant transpiration, but also for detecting and differentiating fungal pathogens.

Cell-Membrane Immunotherapy Based on Natural Killer Cell Membrane Coated Nanoparticles for the Effective Inhibition of Primary and Abscopal Tumor Growth.

Developing effective immunotherapies with low toxicity and high tumor specificity is the ultimate goal in the battle against cancer. Here, we reported a cell-membrane immunotherapy strategy that was able to eliminate primary tumors and inhibited distant tumors by using natural killer (NK) cell membrane cloaked photosensitizer 4,4',4'',4'''-(porphine-5,10,15,20-tetrayl) tetrakis (benzoic acid) (TCPP)-loaded nanoparticles (NK-NPs). The proteomic profiling of NK cell membranes was performed through shotgun proteomics, and we found that NK cell membranes enabled the NK-NPs to target tumors and could induce or enhance pro-inflammatory M1-macrophages polarization to produce antitumor immunity. The TCPP loaded in NK-NPs could induce cancer cell death through photodynamic therapy and consequently enhanced the antitumor immunity efficiency of the NK cell membranes. The results confirmed that NK-NPs selectively accumulated in the tumor and were able to eliminate primary tumor growth and produce an abscopal effect to inhibit distant tumors. This cell-membrane immunotherapeutic approach offers a strategy for tumor immunotherapy.

[In vitro killing activity of the fractional proteins from microtus fortis serum to Schistosoma japonicum juveniles].

OBJECTIVE: To explore the killing effect of different fractional proteins from Microtus fortis (Mf) serum to S. japonicum juveniles, and to find possible association of the proteins with the natural resistance to schistosome infection. METHODS: The proteins in Mf serum were separated by means of ion-exchange column chromatography and molecular sieve column chromatography. After desalted by dialysis and lyophilized, the proteins were dissolved in DMEM medium which contained 300 U/ml penicillin and 300 microg/ml streptomycin, and the two-day old schistosomula were added in for in vitro cultivation (100 +/- 20/well). The killing activity of the fractional proteins to the juvenile worms was defined by mortality rate. RESULTS: 58 fractional proteins were separated from Mf serum, in which six proteins were confirmed to have a significant killing activity to schistosomula. The mortality of schistosomula all reached 37% and above, and the highest mortality (87.5%) was observed in the fraction 18.1, while the negative control was 25.09% (P < 0.01). CONCLUSION: Some fractional proteins in Microtus fortis serum show an effect in the natural resistance to schistosome infection.

[Comparison of specific genomic DNA fragment between Microtus fortis calamorum and Microtus fortis fortis].

Microtus fortis(Taxonomy ID: 100897), also named as reed vole, is classified as Microtus, Micotinae, Cricetidae, Rodentia, Mammalia on taxonomy. Microtus fortis mainly distributes in China. Some areas of Russia, North Korea and Mongolia close to Northeast borderland of China also have a small number of Microtus fortis in distribution. Microtus fortis in China has principally 4 subspecies, and most of them live is the drainage area of Yangtse River. Schistosoma japonicum (one of commonly parasites in China) can infect about 40 kinds of mammalian animals, including the human being, but could not infect Microtus foris. It is known as the only animal in Dongting Lake region of China which has the ability of natural resistance to Schistosoma japonicum. The Microtus fortis domesticated in laboratory has the same biological characteristics as the wild one and these characteristics could be inherited to its progeny steadily. We got a specific DNA fragment from genomic library of Microtus fortis. This DNA fragment in genomic DNA of human beings, Kunming mice, Balb/c mice and C57BL/6J mice could not be detected by dot blot hybridization and PCR, apart from genomic DNA of Microtus fortis. In this report, the differences of genomic DNA in 34 Microtus fortis were compared between Microtus fortis calamorum(Dongting Lake region of southern China) and Microtus fortis fortis (Ningxia province of northern China). The residing localion of these two subspecies is far away about 1,200 kilometers from each other. The genomic DNA of Microtus fortis calamorum and Microtus fortis fortis were extracted and amplified by PCR according to the specific genomic DNAs sequence of Microtus fortis reported previously (Accession number in GenBank: AF277394). The amplified DNA fragments were inserted into pGEM-T easy vector and sequenced. The DNA fragment sequencing results from the two subspecies were compared to detect whether there was any difference. 19 alleles were found from Microtus fortis (20 of Microtus fortis calamorum and 14 of Microtus fortis fortis). The results of multiple sequence alignment showed that 25 single nucleotide polymorphism (SNP) sites were existed in the different Microtus fortis individuals, including transition (G-->A,A-->G,T-->C,C-->T), transversion(G-->T,A-->T,T-->A,C-->A), insertion (CA) and deletion (TGTTTT). The difference of genomic DNA from two subspecies were obvious, especially on the sites of 146, 192, 223, 224 and 235. The insertion of CA on the sites of 223 and 224 as well as A-->G transition on the site of 235 was only occurred in Microtus fortis fortis. They are not found in Microtus fortis calamorum so far. They could be divided into two groups according to phylogenetic tree analysis results, one was the genomic DNA of Microtus fortis calamorum and the other one was that of Microtus fortis fortis. However, the homologues reached up to 98% between two subspecies. These results are very important for us to further understand the genetic background, biological characteristics, evolutionary rule and the anti-schistosoma japonicum mechanism of Microtus fortis at the molecular levels. The specific base changes of the DNA fragment between the two subspecies are probably correlative with the animal immigration, survival conditions, and species evolution.

Identification of Novel Regulatory Elements of Human Stem Cell Factor Gene in MCF Cell.

Human stem cell factor(hSCF)is a pluripotent growth factor that regulates proliferation, differentiation and migration of certain mammalian stem cells, such as primordial germ cells etc. It is shown that hSCF and its receptor are commonly co-expressed in human breast cancer cells. Up to now, the definite regulatory mechanism of hSCF gene in breast cancer cells is unclear, except that its 5'flanking sequence contains essential elements for regulating transcription. To localize the regulatory elements responsible for the regulation of the hSCF gene, we performed transient transfection study in MCF cells, with a series of luciferase reporter gene constructs, containing different 5x end deletions of hSCF gene. This study indicates that the region of -1190 -853 significantly enhanced the luc gene expression, while the region of -339 -162 inhibited the expression. Eletrophoretic mobility shift assay confirmed that MCF nuclear extract proteins bound to both -1190 -853 and -339 -273 regions, forming specific DNA-protein complexes, indicating that there were nuclear protein binding sites in these regions. The results suggest that both -1190 -853 and -339 -273 DNA fragments of the hSCF 5'flanking sequence may be novel regulatory elements, and may play a role in the regulation of hSCF gene expression in MCF cells.