17 results about "Cell plasma membrane" patented technology

The invention provides a preparation method of dendritic mesoporous silicon bionic nanoparticles loaded with RNA (Ribonucleic Acid) molecules, which comprises the following steps: preparing dendritic mesoporous silicon nanoparticles DMSNs by adopting a double-phase layering method, dissolving a surfactant CTAC and a catalyst TEA in water to serve as a water phase solution, dissolving ethyl silicate TEOS in cyclohexane to serve as an oil phase, slowly dropwise adding the oil phase into the water phase, and reacting for 2-4 hours to obtain the dendritic mesoporous silicon bionic nanoparticles loaded with the RNA molecules. The DMSNs nanoparticles are obtained; amino groups are introduced to the surfaces of DMSNs through gamma-aminopropyltriethoxysilane APTES so as to enhance electrostatic attraction to nucleic acid, and after a surfactant template is removed, the DMSNs are mixed with RNA molecules in ribozyme-free water; meanwhile, a cytoplasmic membrane is harvested by using a hypotonic method, and the dendritic mesoporous silicon bionic nanoparticles loaded with the RNA molecules are prepared through ultrasonic treatment and polycarbonate film extrusion. The method is simple and convenient, and has a wide application prospect in RNA delivery.

The invention belongs to the technical field of bioengineering, and discloses a plasma membrane localized green fluorescence overexpression vector as well as a construction method and application thereof. The cytoplasmic membrane localized green fluorescence overexpression vector contains an expression vector skeleton and an AtCBL1 gene connected to the expression vector skeleton, wherein the nucleotide sequence of the AtCBL1 gene is shown as SEQIDNO: 1, and the expression vector skeleton also carries a fluorescent protein gene. The construction method of the vector comprises the following steps: S1, carrying out enzyme digestion and recovery on an AtCBL1 gene sequence with a nucleotide sequence as shown in SEQIDNO: 1 obtained by amplification; s2, performing same enzyme digestion on the expression vector carrying the fluorescent protein gene, and recovering; and S3, connecting the AtCBL1 gene subjected to enzyme digestion with the expression vector to form a recombinant plasmid, converting a connection product into an escherichia coli competent cell, and extracting the recombinant plasmid from positive clone to obtain the plasma membrane localized green fluorescence overexpression vector.

The present invention identifies the expression mechanism of an RAB10 or MYH9 protein in a cell plasma membrane of a patient having resistance to a pancreatic cancer therapeutic agent. The present invention provides an RAB10 or MYH9 protein as a biomarker for diagnosing the resistance of a pancreatic cancer patient to a therapeutic agent, and provides a pharmaceutical composition for mitigating resistance to a therapeutic agent for pancreatic cancer through the use of an anti-oligonucleotide that inhibits the RAB10 or MYH9 protein.

Delivery of a payload across a cytoplasmic membrane includes providing a population of cells, contacting the population of cells with a volume of an aqueous solution. The aqueous solution includes an effective load and an alcohol at a concentration greater than 5%. The volume of the aqueous solution may be a function of the exposed surface area of the cell population, or a function of the number of cells in the cell population. Related compositions, instruments, systems, techniques, and articles are also described herein.

Crimean-Congo hemorrhagic fever virus (CCHFV) is an important human pathogen. Limited evidence suggests that antibodies can protect humans against lethal CCHFV disease, but the protective efficacy of antibodies has never been evaluated in adult animal models. Here adult mice were used to investigate the protection provided by glycoprotein-targeting neutralizing and non-neutralizing monoclonal antibodies (mAbs) against CCHFV infection. A single non-neutralizing antibody (mAb-13G8) was identified that protected adult type I interferon deficient mice >90% when treatment was initiated prior to virus exposure and >60% when administered after virus exposure. Neutralizing antibodies known to protect neonatal mice from lethal CCHFV infection, failed to confer protection regardless of IgG subclass. The target of mAb-13G8 was identified as GP38, one of multiple proteolytically-cleaved glycoproteins derived from the CCHFV glycoprotein precursor polyprotein. Robust protection required complement activity, but not Fc-receptor functionality. Consistently, it was found that GP38 previously identified as a secreted molecule also localizes to viral envelope and cellular plasma membranes. This study reveals GP38 as an important antibody target for CCHFV and lays the foundation to develop novel vaccines and immunotherapeutic against CCHFV in human.

The invention relates to the technical field of cell biology, in particular to a non-cell stimulant for amplifying NK cells derived from pluripotent stem cells. The invention provides an extraction method of an aAPC cell membrane. According to the extraction method, the amplification efficiency of aAPC on NK cells can be remarkably improved through specific cell factors or a combination of the specific cell factors; after the cells are subjected to hypotonic treatment, the cells are filtered by a centrifugal column, and the extracted cytoplasmic membrane (PM) component does not have living cells and is high in safety when being used for NK (Natural Killer) cell amplification; the PM film extraction process is simple and efficient, PM film preparation can be performed in a common laboratory with a high-speed refrigerated centrifuge, and large-scale preparation is easy; by means of the scheme (the hypotonic solution and the centrifugal tubular column), the PM extraction yield is high, 1500-2000 micrograms of PM component protein can be obtained through 5.0 E7 cells, and the whole process is short in consumed time (about 45 min can be completed).

This disclosure provides a method and apparatus for identifying enhancer activity in live cells, enabling sensitive detection of enhancer activity within live cells. The method utilizes a nano-electroporation biochip to perform highly focused perforation of the cell membrane and provides the driving force to accelerate the delivery of enhancer probes to the cell nucleus. The enhancer probes are designed using a cyclic amplification strategy, increasing the fluorescence signal by more than 100-fold within one hour. In the embodiments, the method and apparatus successfully detected the activity of CCAT1 enhancer in live cells without affecting cell viability, demonstrating the robustness and reliability of the proposed method and apparatus in detecting enhancer activity in live cells. This disclosure provides a convenient and universal platform for research on enhancer gene regulatory mechanisms in live cells and for clinical diagnostic applications.

The invention relates to the technical field of fluorescent probes, and discloses a preparation method and application of a cytoplasmic membrane covalent labeling fluorescent probe based on non-membrane permeability, and the preparation method comprises the following steps: S1, synthesizing 4-(2, 3, 3-trimethyl-3H-indole-1-onium-1-yl) butane-1-sulfonate; s2, synthesizing a 2-halogen substituted N-((1E, 2Z)-3-9 (phenylamino) allylidene) phenylamine derivative; and S3, synthesizing a target product, namely the Sulfo-Cy5-X fluorescent probe. The novel cytoplasmic membrane fluorescent probe is prepared by adopting a membrane impermeability-based cytoplasmic membrane covalent labeling principle and introducing a sulfonic acid group with high water solubility and a covalent reaction site, so that a long-time and specific labeling function on the cytoplasmic membrane is realized; according to the method, the problems of short imaging time and low water solubility of a traditional fluorescent probe are solved, and a more ideal tool is provided for in-situ observation of dynamic change of the cytoplasmic membrane.

The invention discloses a pediococcus pentosaceus D17-sourced cytoplasmic membrane vesicle and application thereof, and belongs to the technical field of microorganisms. The pediococcus pentosaceus D17 with excellent probiotic characteristics is separated from fresh excrement of Lichuan horses for the first time, the preservation number of the pediococcus pentosaceus D17 is CCTCC (China Center For Type Culture Collection) NO: M20252051, and meanwhile, the application of the pediococcus pentosaceus D17 and the cytoplasmic membrane vesicles of the pediococcus pentosaceus D17 in resisting salmonella infection is researched. Experimental verification shows that the separated and screened pediococcus pentosaceus D17 and the cytoplasmic membrane vesicles thereof have the effect of resisting salmonella infection and can promote expression of tight junction protein, and the treatment effect of the cytoplasmic membrane vesicles derived from the pediococcus pentosaceus D17 is superior to that of the pediococcus pentosaceus D17. Due to the fact that the cytoplasmic membrane vesicles are prepared through centrifugation of bacterial liquid supernatant, the cytoplasmic membrane vesicles are safer and free of drug resistance risks, and a new thought and a new approach are provided for application of the cytoplasmic membrane vesicles sourced from pediococcus pentosaceus to preparation of salmonella infection resisting products.

The invention discloses a method for detecting response sensitivity of von Willebrand Factors (von Willebrand Factors, von Willebrand Factors, von Willebrand Factors, von Willebrand Factors, von Willebrand Factors, von Willebrand Factors, von Willebrand Factors, von Willebrand The method comprises the following steps: firstly, inserting an FRET pair at two sides of an A2 structural domain of a full-length vWF to construct an FRET-based vWF biosensor; transfecting the vWF biosensor into cells, so that the vWF biosensor is anchored on the surface of a cytoplasmic membrane; the cells are inoculated in a fluid experiment shearing device, and an FRET microscopic imaging system collects image data of ECFP and FRET imaging channels of the cells under the action of fluid shearing force; performing data processing and analysis on the image data; and analyzing the response sensitivity of the vWF under the action of the micro shear force. The constructed biosensor and the analysis method thereof can be used for screening research of drugs for treating hemophilia.

The application discloses a water-soluble AIE fluorescent probe, and a structure formula is shown in the following formula (I), wherein R1 and R2 are independently selected from alkyl alcohol with carbon number of 1-6; X is selected from water-soluble anions, and M is selected from water-soluble cations. The fluorescent probe has AIE characteristics, can realize rapid and efficient staining of cytoplasmic membranes, and does not need to be cleaned; the fluorescent probe has good water solubility, small cytotoxicity, and can be used in in vivo cell fluorescence imaging; the fluorescent probe has orange red fluorescence color, large Stokes shift, and strong anti-self-fluorescence interference capability.

The invention discloses a high-affinity anti-human and monkey PCSK9 antibody, and belongs to the field of biological medicine. The anti-human PCSK9 antibody comprises a heavy chain variable region of which the sequence is SEQ ID NO.13 and a light chain variable region of which the sequence is SEQ ID NO.14, or a heavy chain variable region of which the sequence is SEQ ID NO.15 and a light chain variable region of which the sequence is SEQ ID NO.16. The six anti-human PCSK9 antibodies are obtained by immunizing mice with human PCSK9 protein and combining with a hybridoma cell line and screening, and can be combined with human PCSK9 with high affinity and high selectivity, so that the combination of the human PCSK9 and LDLR (Low Density Lipoprotein Receptor) on a cytoplasmic membrane is blocked, and the degradation of the LDLR is reduced. PCSK9 is highly expressed in tumors, tumor immune recognition and CD8 + T cell anti-tumor activity are inhibited, and the PCSK9 participates in tumor immune escape. The anti-human PCSK9 antibody provided by the invention can provide candidate antibody molecules for tumor immunotherapy.

The invention relates to the technical field of fluorescent probes, in particular to a preparation method and application of a plasma membrane targeted heavy atom-free photosensitizer. Ferroptosis is an iron-dependent programmed cell death form caused by abnormal accumulation of lipid peroxidation (LPO) on a cell membrane and has important treatment potential in the aspects of overcoming tumor drug resistance and metastasis, and photodynamic therapy (PDT) is an ideal method for inducing the LPO to cause the ferroptosis. The invention provides a photosensitizer Mem-ANSiR capable of specifically targeting a cytoplasmic membrane, the photosensitizer can efficiently generate < 1 > O2 and O2 at the same time through I-type and II-type reactions, and a ferroptosis pathway is effectively promoted through accumulation of lipid peroxide. It is worthy of notice that the Mem-ANSiR can still efficiently kill cancer cells in a low-oxygen microenvironment, and a brand new photodynamic tool is provided for solid tumor treatment.

This invention relates to a method for increasing the permeability of a cell membrane by directing at least one cell to or near a structure containing particles capable of absorbing electromagnetic radiation. These particles are present at a concentration of 0.001 vol% to 20 vol% and are embedded in the material of the structure. At least 60% of the particles present in the structure are embedded in the material such that the shortest distance L between these particles and the free surface S of the structure is 1 nm to 500 nm. The invention also relates to a structure suitable for permeating cells during photothermal processes and the use of said structure.

The application discloses a water-soluble cationic fluorescent probe dye and application thereof in cell membrane staining. The fluorescent probe has good water solubility, can be used in in-vivo cell fluorescence imaging, realizes rapid and efficient staining of cell plasma membranes, and does not need to be cleaned. The fluorescent probe has an orange red fluorescence color, has a large Stokes shift, and has strong anti-self-fluorescence interference capability.

The present invention discloses an application of a willow natural resistance-related macrophage protein SsNramp1 in genetic engineering for cadmium-contaminated soil remediation. The willow SsNramp1 in the present invention has the activity of transporting cadmium, and SsNramp1 is a membrane transport protein located on the cytoplasmic membrane. After SsNramp1 is activated to express the protein in tobacco using Agrobacterium tumefaciens-mediated genetic transformation technology and a 35S strong promoter, the growth of tobacco transfected with the SsNramp1 gene is not significantly affected under a cadmium exposure environment compared with the wild type, while the cadmium concentration in the root system increases by 64%-128%, and the cadmium concentration in the aboveground part increases by 35%-51%. The present invention illustrates that willow SsNramp1 is of great significance in cadmium-contaminated soil remediation engineering.

The invention relates to a method to increase the permeability of the plasma membrane of cells by introducing at least one cell on or near a structure comprising particles able to absorb electromagnetic radiation. The particles, present in a concentration ranging between 0.001 vol % and 20 vol %, are embedded in the material of the structure. At least 60% of the particles present in the structure are embedded in the material in such a way that the shortest distance L between these particles and the free area surface S of the structure ranges between 1 nm and 500 nm.The invention further relates to a structure suitable for use in a photothermal process to permeabilize cells and to the use of such structure.