199 results about "Cytoplast" patented technology

Adjuvant combinations comprising at least one microbial TLR agonist such as a whole virus, bacterium or yeast or portion thereof such a membrane, spheroplast, cytoplast, or ghost, a CD40 or 4-1BB agonist and optionally an antigen wherein all 3 moieties may be separate or comprise the same recombinant microorganism or virus are disclosed. The use of these immune adjuvants for treatment of various chronic diseases such as cancers and HIV infection is also provided.

Chimeric receptor genes suitable for endowing lymphocytes with antibody-type specificity include a first gene segment encoding a single-chain Fv domain of a specific antibody and a second gene segment encoding all or part of the transmembrane and cytoplasmic domains, and optionally the extracellular domain, of an immune cell-triggering molecule. The chimeric receptor gene, when transfected to immune cells, expresses the antibody-recognition site and the immune cell-triggering moiety into one continuous chain. The transformed lymphocytes are useful in therapeutic treatment methods.

The present invention includes nuclear fertility restorer genes, proteins encoded by those genes and transgenic plants and plant cells containing those genes. More particularly, the nuclear fertility restorer genes can be used to restore fertility in cytoplasmic male-sterile plants such as Brassica napus. Preferably, the nuclear fertility restorer genes are used with the Ogura (ogu) CMS system in Brassica napus.

The present invention includes nuclear fertility restorer genes, proteins encoded by those genes and transgenic plants and plant cells containing those genes. More particularly, the nuclear fertility restorer genes can be used to restore fertility in cytoplasmic male-sterile plants such as Brassica napus. Preferably, the nuclear fertility restorer genes are used with the Ogura (ogu) CMS system in Brassica napus.

The invention discloses functional albumin and a preparation method of a nano preparation of the functional albumin. The nano preparation of the functional albumin consists of the functional albumin, metal ions and a drug; the metal ions can simultaneously form coordination bonds with the functional albumin and the drug, and can form nanoparticles through induced self-assembly. The nano preparation, through an endocytosis mediated by an albumin receptor (SPARC) on the surface of tumor cells, can deliver the drug into the drug-resistant tumor cells, so as to effectively avoid the exocytosis effect of a p-gp pump on the drug, and then as the coordination bonds break in an acid tumor cell environment through a pH responsibility, the drug releases in cytoplast, enters cell nucleus and inlays in DNA so as to inhibit the synthesis of nucleic acid; and therefore, the growth of the tumor cells is inhibited. Through in vitro characterization, the nano preparation can achieve the relatively good pH responsibility; and through activity evaluation on a cellular level, the system is capable of effectively delivering the drug into the cells, so as to achieve relatively good pH responsive release.

The invention relates to a breeding method and application of a cytoplasmic male sterility restoring line of brassica napus rapeseed and radish. The method comprises the following steps of: choosing rapeseed S1628 with the preservation number of CCTCC NO: P201203, performing continuous inbreeding directional selection for six years to breed an excellent, high-oil and stable inbred line; using the cytoplasmic male sterility restoring line of the brassica napus rapeseed and radish as a female parent and using the bred excellent and stable inbred line as a male parent to select out strains comprising radish cytoplasmic male sterility restoring genes from large amounts of filial generations; using the strains comprising the radish cytoplasmic male sterility restoring genes as the female parent and using Zhongshuang No.2, Huashuang No.3, Huyou 17, Zunyou No.1 and Zheyou 758 of double-low winter rapeseed varieties in China as recurrent male parents to hybridize, performing the continuous inbreeding directional selection for six years to breed the cytoplasmic male sterility restoring line of the double-low and high-oil brassica napus rapeseed and radish. The restoring line has been crossed with hybrid varieties. A gap of producing the hybrid varieties by the cytoplasmic male sterility restoring line of the double-low radish in China is filled, and the application prospect is broad.

The invention relates to the field of accessories and preparations with medicinal functions, in particular to a kind of oligopeptide-based pH-sensitive amphoteric ion lipid (I) or (II). In the invention, the derivatives of the oligopeptide amphoteric ion lipid have different surface charges under different pH conditions and can take relatively strong negative charges in a physiological environment with pH near neutrality so that the lipid has good blood compatibility; when the lipid reaches a tumor part, the surface charges of the oligopeptide amphoteric ion lipid are reversed in an acidulous tumor environment, so that the original electronegativity is reversed into electropositivity; and a carrier with positive charges is easily combined with the surface of a tumor cell, and the tumor cell targeting can be effectively realized through an endocytosis path. In lysosome, a cationic carrier continuously realizes a function of proton sponge or a function of splitting lysosome membranes, and the two functions can ensure the integrity of the carrier, so that the carrier can safely escape to cytoplast or other organelles.

This disclosure concerns high-resolution mapping and candidate gene cloning of Rf4, a maize restorer of fertility gene that restores fertility to C-type cytoplasmic male sterility. The disclosure also relates to molecular markers that are tightly-linked to, or reside within, the Rf4 gene. In some embodiments, methods are provided whereby hybrid seeds may be produced from crosses of a male plant comprising nucleic acid molecular markers that are linked to or that reside within the Rf4 gene and a female plant carrying C-type CMS.

The invention relates to a method of making pluripotent stem cells that does not involve the formation of early preimplantation embryos or fetal tissue. The method has general utility in the production of pluripotent stem cells from many mammalian species but has particular application in man where pluripotent stem cell production can be customized to particular human individual. The method involves the fusion of donor somatic or stem cells (or their karyoplasts) with cytoplasmic, membrane-delimited fragments of mammalian oocytes or zygotes. After the initial genomic reprogramming occurs, the cells can proliferate and thus multiply in vitro yielding a large number of autologous cells for cell therapy application. The result of this process is a cell population genomically identical to the somatic, differentiated cells derived from an individual patient. However, these cells are pluripotent in that upon application of specific growth factors, the cells are capable of differentiating into specific cell types as required by the sought clinical indication.

Methods for producing stem cell banks, preferably human, which optionally may be transgenic, e.g., comprised of homozygous MHC allele cell lines are provided. These cells are produced preferably from parthenogenic, IVF, or same-species or cross-species nuclear transfer embryos or by dedifferentiation of somatic cells by cytoplasm transfer. Methods for using these stem cell banks for producing stem and differentiated cells for therapy, especially acute therapies, and for screening for drugs for disease treatment are also provided.

The present invention provides the nucleotide sequence and encoded amino acid polypeptide sequence of one cytoplamis male sterile restoring gene OsRf1 originating from rice. The gene is member of PPR gene family, and is one constitutive expression gene. The present invention also relates to the application of utilizing the gene in transforming rice and other plant breeding restorer and the application of the molecular label produced based on the gene sequence in breeding.

A method of obtaining a mixture of cells enriched in hepatic progenitors is developed which comprises methods yielding suspensions of a mixture of cell types, and selecting those cells that are classical MHC class I antigen(s) negative and ICAM-1 antigen positive. The weak or dull expression of nonclassical MHC class I antigen(s) can be used for further enrichment of hepatic progenitors. Furthermore, the progenitors can be selected to have a level of side scatter, a measure of granularity or cytoplasmic droplets, that is higher than that in non-parenchymal cells, such as hemopoietic cells, and lower than that in mature parenchymal cells, such as hepatocytes. Furthermore, the progeny of the isolated progenitors can express alpha-fetoprotein and / or albumin and / or CK19. The hepatic progenitors, so isolated, can grow clonally, that is an entire population of progeny can be derived from one cell. The clones of progenitors have a growth pattern in culture of piled-up aggregates or clusters. These methods of isolating the hepatic progenitors are applicable to any vertebrates including human. The hepatic progenitor cell population is expected to be useful for cell therapies, for bioartificial livers, for gene therapies, for vaccine development, and for myriad toxicological, pharmacological, and pharmaceutical programs and investigations.

The invention discloses an umbilical cord blood Treg cell in-vitro amplification method based on trophoblastic cells and application. The specific technical method comprises the steps that firstly, umbilical cord Wharton's jelly mesenchymal stem cells are adopted as the trophoblastic cells to induce preliminary proliferation of Treg cells in umbilical cord blood mononuclear cells; then, pure Tregcells are obtained through magnetic bead sorting; and finally, the Treg cells are stimulated to be rapidly amplified by using optimized amplification factors. According to the amplification method, human AB plasma, IL-2, rapamycin, an RARA agonist and a DNA methyltransferase inhibitor are used as the optimized amplification factors, and a large number of umbilical cord blood Treg cells with high purity and high activity can be prepared within two weeks. Umbilical cord blood is used as a raw material for Treg cell amplification, batch preparation can be achieved, and Treg cell quality fluctuation caused by individual differences of samples can be reduced. The umbilical cord blood Treg cells have low immunogenicity and can be used as universal cells for clinical research, such as autoimmunediseases, graft-versus-host diseases and the like.

The invention provides a method for identifying soybean RN type cytoplasmic male sterile line seed purity through a molecular marker. The method comprises the steps of: extracting genomic DNA of an RN type cytoplasmic male sterile line seed; carrying out PCR (Polymerase Chain Reaction) amplification by taking the genomic DNA as a template and utilizing a primer InDel-cms1, and carrying out gel electrophoresis analysis on an amplification product, wherein the length of a sterile line fragment after amplification is 200bp, and the length of a maintainer line fragment after amplification is 212bp; and accurately distinguishing the sterile line and the maintainer line through the length difference of the fragments, wherein the seed with the amplified fragment different from the amplified sterile line fragment is a false sterile line seed, and the percentage of the number of sterile line fragments with specific length obtained by detection accounting for all the detected seeds is the purity of RN type cytoplasmic male sterile line seeds. The method can be used for rapidly, accurately and reliably identifying the maintainer line mixed in the RN type soybean cytoplasmic male sterile line by a molecular biology experiment means so as to ensure the purity of sterile line seeds.

In accordance with the present invention, it has been discovered that glucose and incretin hormones promote pancreatic islet cell survival via the calcium and cAMP dependent induction, respectively, of the transcription factor CREB. Specifically, a signaling module has been identified which mediates cooperative effects of calcium and cAMP on islet cell gene expression by stimulating the dephosphorylation and nuclear entry of TORC2, a cytoplasmic CREB coactivator. The module comprises a cAMP regulated snf1-like kinase called SIK2 and the calcium regulated phosphatase calcineurin, both of which associate with TORC2 in the cytoplasm. TORC2 is repressed under basal conditions through a phosphorylation dependent interaction with 14-3-3 proteins. cAMP and calcium signals stimulate CREB target gene expression via complementary effects on TORC2 dephosphorylation; cAMP disrupts TORC2-associated activity of SIK2 or related family members, whereas calcium induces TORC2 dephosphorylation via calcineurin. These findings provide a novel mechanism by which CREB activates cellular gene expression, depending on nutrient and energy status, and facilitate development of assays to identify compounds which modulate the role of TORCs. In accordance with the present invention, it has been discovered that fasting and energy-sensing pathways regulate the gluconeogenic program in liver by modulating the nuclear entry of a transcriptional coactivator called Transducer of Regulated CREB Activity 2 (TORC2). Hepatic TORC2 over-expression induces fasting hyperglycemia, whereas knockdown of TORC2 leads to fasting hypoglycemia and silencing of the gluconeogenic program. Since a majority of individuals with Type II diabetes exhibit fasting hyperglycemia due to elevated hepatic gluconeogenesis, compounds that enhance TORC2 phosphorylation will find use as therapeutic agents in this setting.

This invention provides a nucleus transplantation method for mammal somatic cells characterizing in taking the oocyte of cross-fertilized F1 generation of the mammal as the recipient somatic cell for nucleus transplantation, which can increase the re-programming ability of recipient cytoplast of nucleus transplantation of mammals obviously, improve the growth of re-structured embryo to increase the clone efficiency of somatic cells.

The present disclosure provides a method for selecting a plant comprising a functional restorer gene for maize S-type cytoplasmic male sterility comprising the steps of (a) screening a population of plants for at least one marker nucleic acid, wherein the marker nucleic acid comprises an allele linked to the functional restorer gene for maize S-type cytoplasmic male sterility; (b) detecting the marker nucleic acid; (c) identifying a plant comprising the marker nucleic acid; and (d) selecting the plant comprising the marker nucleic acid, wherein the plant comprising the marker nucleic acid further comprises the functional restorer gene for maize S-type cytoplasmic male sterility. The present disclosure also provides methods for restoring fertility in a progeny of an S-type cytoplasmic male sterile plant and methods for transferring an Rf3 gene into a progeny plant.

The invention belongs to the technical field of molecular biology and discloses a pepper sterility restoring gene closely linked molecular marker, a method and application. The molecular marker is scd06-17 and comprises a nucleotide sequence shown as SEQ ID NO.1 and a nucleotide sequence shown as SEQ ID NO.2. The molecular marker is stable and reliable in result and is a co-dominant molecular marker, whether a pepper material contains a cytoplasmic male sterility restoring gene or not can be determined in a pepper steeling stage, and accordingly breeding efficiency is improved, and a breeding process is accelerated. Compared with other domestic and international published pepper restoring gene molecular markers, the pepper sterility restoring gene closely linked molecular marker is significant to pepper three-line hybrid seed production and restoring material molecular marker assisted selection breeding.

The invention discloses a breeding method of wavy Brassica oleracea var. acephala f. tricolor Hort., which comprises the following steps: hybridizing by using asparagus broccoli cytoplast male sterile material CMS034 as a female parent and red curly kale inbred line Ruiyu 6 as a male parent; backcrossing with the original male parent by selecting the individual plant, which has characters closest to the parental characters, from filial generation as the female parent; repeating the second step, and continuously backcrossing to obtain a red curly kale male sterile line; and hybridizing to obtain the wavy Brassica oleracea var. acephala f. tricolor Hort. hybrid by using the red curly kale male sterile line as the female parent and amaranth broad-leaf Brassica oleracea var. acephala f. tricolor Hort inbred line Y305 as the male parent. The new hybrid species obtained by the method disclosed by the invention has the advantages of high ornamental value, long ornamental period, cold resistance and disease resistance.

The invention belongs to the mammal reproduction technology field, particularly relates to a method for obtaining cloned embryos with cow somatic cells and discloses a method for producing cow cloned embryos simply and efficiently. According to the research results that pig somatic cells dissolves and disappears automatically in thirty minutes after being injected into ooplasm, whole cells with sound membrane are extracted from cows as donors and are injected into the whole cell cytoplast directly to construct cow cloned embryos, simplifying the operational process of somatic cell nuclear transfer. The method is characterized in that: (1) selecting and cultivating cow ovarian cumulus- oocytes complex; (2) enucleating of the occytes; (3) preparing donor cells; (4) preparing co-cultivation cell monolayer; (5) constructing reconstructed fetus; (6) activating the reconstructed fetus; and (7) cultivating the reconstructed fetus. The method has the best feather of easy operation, simple steps, stable results, providing a new idea and method for the expansion and the further study of cow somatic cells nuclear transplanting technology.

The present invention relates to a method for inhibiting intracellular activated RAS using an intact immunoglobulin-type antibody having the ability to penetrate the cytosol. The present invention also relates to a heavy-chain variable region (VH) which induces an intact immunoglobulin-type antibody to actively penetrate the cytosol of living cells through endocytosis and endosomal escape and to bind to activated RAS in the cytosol, and to an antibody comprising the same. The present invention also relates to a method of inhibiting the growth of cancer or tumor cells and a method of treating cancer or tumor, by use of the antibody. The present invention also relates to a method for screening a heavy-chain variable region that binds specifically to RAS in the cytosol. The present invention also relates to a biologically active molecule fused to the antibody and selected from the group consisting of peptides, proteins, small-molecule drugs, nanoparticles and liposomes. The present invention also relates to a composition for prevention, treatment or diagnosis of cancer, comprising: the antibody; or a biological active molecule fused to the antibody and selected from the group consisting of peptides, proteins, small-molecule drugs, nanoparticles and liposomes. The present invention also relates to a polynucleotide that encodes the light-chain variable region and the antibody.According to the present invention, the method for inhibiting intracellular activated RAS using an intact immunoglobulin-type antibody having the ability to penetrate the cytosol is achieved by allowing the antibody to penetrate living cells and to specifically recognize activated (GTP-bound) RAS in the cytosol. Thus, the antibody can target activated (GTP-bound) RAS in the cytosol of living cells and inhibit the activity of the RAS.Furthermore, the antibody light-chain variable region according to the present invention and an antibody comprising the same is able to penetrate living cells and localize in the cytosol, without having to use a special external protein delivery system. Particularly, the antibody light-chain variable region according to the present invention can easily interact with various human heavy-chain variable regions (VHs) and has the ability to penetrate the cytosol and remain in the cytosol, and an intact IgG-type monoclonal antibody comprising the light-chain variable region can penetrate cells and localize in the cytosol, and shows no cytotoxicity non-specific for target cells.The antibody heavy-chain variable region according to the present invention and an antibody comprising the same can selectively inhibit mutations of the major drug resistance-related factor RAS of conventional various tumor therapeutic agents, and can exhibit synergistic anticancer activity with conventional therapeutic agents. In addition, the cytosol-penetrating, intact immunoglobulin-type antibody according to the present invention can penetrate cells and remain in the cytosol, without affecting the high specificity and affinity of a human antibody heavy-chain variable region (VH) for antigens, and thus can localize in the cytosol which is currently classified as a target in disease treatment based on small-molecule drugs, and at the same time, can exhibit high effects on the treatment and diagnosis of tumor and disease-related factors that show structurally complex interactions through a wide and flat surface between protein and protein.

The invention relates to a preparation method of amycin controlled-release chitosan nano particles with pH / oxido-reduction dual response. Thiolated chitosan, carboxymethyl chitosan, sodium tripolyphosphate, doxorubicin hydrochloride and hydrogen peroxide are taken as raw materials. The preparation method comprises the following steps: mixing a thiolated chitosans solution with a doxorubicin hydrochloride solution so as to obtain a solution I; mixing a carboxymethyl chitosan solution with a sodium tripolyphosphate solution so as to obtain a mixed solution II; dropwise adding the mixed solution I into the mixed solution II while stirring, controlling the dosage of carboxymethyl chitosan and thiolated chitosan, regulating the ph value of the solution to 6-8, performing ultrasonic blending, performing ion crosslinking and polymer condensing, then dropwise adding hydrogen peroxide into the system so as to finish oxidization crosslinking, performing room-temperature stirring reaction, separating products, and drying the products to obtain the amycin controlled-release chitosan nano particles with pH / oxido-reduction dual response. The amycin controlled-release chitosan nano particles with pH / oxido-reduction dual response is capable of escaping away from tumor cell endosome / lysosome, is capable of automatically regulating and controlling to realize cytoplast release based on unique microenvironments of tumor cells, improving the amycin medicine delivery efficiency and the medical effect, and has a favorable research and development application background in multiple aspects such as medicine and medical materials.

The invention relates to a method for transforming a sterile line of Chinese kale by utilizing vernalization and generation adding. The method comprises the following steps of: carrying out hybridization by taking a male sterile line of broccoli cytoplast as a female parent and a selfing line of the Chinese kale as a male parent, so as to obtain hybridized seeds; after the hybridized seeds are planted in summer, carrying out vernalization on plants, and meanwhile, selecting the sterile plants by utilizing pollen microscopic examination or single-plant bagging selfing; and taking the selfing line of the Chinese kale as a recurrent parent, carrying out 3-5-generation backcross with the recurrent parent and the sterile plants, so as to obtain the male sterile line of Chinese kale cytoplast. According to the method, the sterile line of the Chinese kale is transformed by utilizing the male sterile line of broccoli cytoplast, biological characters of the sterile line of the Chinese kale are stable, stamens degenerate, petals and pistils develop normally, nectaries develop well, and seed pods develop normally; the sterile line can be subjected to generation adding propagation under a normal growth condition of the plants, the transformation process is accelerated, the procedures are simplified, the cost is saved, and the purity of the Chinese kale can be improved.

The invention discloses a molecular marker for cabbage cytoplasm male sterile line restore gene and an application thereof, the molecular marker tightly linked with the cabbage cytoplasm male sterile line gene is the InDe178 maker and restorer sequence SCAR 42 maker positioned at two sides of cabbage CMS 7311 fertility restore gene and tightly linked with the cabbage CMS 7311 fertility restore gene, the genetic distance between the molecular marker InDe178 / SCAR 42 and the fertility restore gene are 0.91cM and 0.92cM respectively, and the fertility restore gene is limited in a range of 337kb of cabbage A09 chromosome. Two molecular markers can be used for molecular marker auxiliary selection of cabbage CMS 7311 fertility restore gene, selection efficiency is increased, breeding process is accelerated; and the molecular markers establish a base for map-based cloning of fertility restore gene, reveal of cytoplasm male sterility and molecular mechanism of fertility restore gene.

The Chinese cabbage hybrid breeding process features that the process includes interspecific crossing with cabbage type rape radish cytoplast male sterile material RC97-1 as female parent and Chinese cabbage self-bred line 1138 as male parent to obtain transfer bred primary generation seed, selecting the excellent single plant as the backcross female parent for the next turn; continuous seven-generation backcross transfer breeding, economic character selection, disease resistance identification to obtain Chinese cabbage cytoplast male sterile material RC7 with stable sterility and economic character the same as that of corresponding maintainer line; and the further crossing with the Chinese cabbage cytoplast male sterile material RC7 as male parent and Chinese cabbage self-bred line 04S245 as male parent to obtain the Chinese cabbage hybrid. The Chinese cabbage hybrid Jingqiu-70, or RC7*04S245, has the features of early ripening, compact plant, high disease resistance, etc.

The invention discloses Yunnan red pear-1 metallothionein gene PyMT1 and a prokaryotic expression vector thereof, and the prokaryotic expression vector is a prokaryotic expression vector which contains the Yunnan red pear-1 metallothionein gene PyMT1. According to the invention, PyMT1 gene is cloned from Yunnan red pear-1; the over expression of the gene in bacterial cells is controlled by T7 promoter and lactose operon with the induction of IPTG; PyMT1 protein is synthesized and secreted into cytoplasm; the expression level is detected by SDS-PAGE; test results show that the expression level of the PyMT1 protein reaches a maximum when the expression is inducted by 1mM IPTG at 37 DEG C for 4 hours and when the expression is inducted by 0.5 mM IPTG at 37 DEG C for 4 hours; the vector adopted by the invention can successfully express a lot of Yunnan red pear-1 metallothionein; the test results also demonstrates that the Yunnan red pear-1 metallothionein gene PyMT1 of Yunnan red pear has a typical function of heavy metal ion tolerance of the metallothionein gene, which lays a foundation for further research of PyMT1 functions and the applications of the metallothionein gene in microbial tolerance to heavy metal.

The present invention provides methods and compositions for enhancing channel activity to the mutant cystic fibrosis trans-membrane conductance regulator protein (CFTR). The compositions of the invention comprise polypeptides containing CFTR sub-domains that are designed to mimic the folding defect of the full length mutant CFTR proteins, resulting in competitive binding to cytoplasmic chaperones such as Hsc / Hsp7O and Hdj2. The methods of the invention comprise transduction, or recombinant expression, of CFTR polypeptides in a cell expressing mutant CFTR. The presence of the CFTR polypeptide results in a dominant effect whereby the CFTR polypeptide competes with the endogenously expressed mutant CFTR for binding to cytoplasmic chaperones such as Hsc / Hsp70 and Hdj2. Mutant CFTR proteins include, but are not limited to, DeltaF508 CFTR. The present invention is based on the discovery that reduced binding of cytoplasmic chaperones to the endogenous DeltaF508 CFTR, mediated by the presence of CFTR polypeptides, results in restoration of plasma membrane localization and channel activity. The methods and compositions of the invention can be used to restore channel activity in cystic fibrosis subjects carrying genetic defects in the CFTR gene, such as for example, DeltaF508 CFTR.

A method of obtaining a mixture of cells enriched in hepatic progenitors is developed which comprises methods yielding suspensions of a mixture of cell types, and selecting those cells that are classical MHC class I antigen(s) negative and ICAM-1 antigen positive. The weak or dull expression of nonclassical MHC class I antigen(s) can be used for further enrichment of hepatic progenitors. Furthermore, the progenitors can be selected to have a level of side scatter, a measure of granularity or cytoplasmic droplets, that is higher than that in non-parenchymal cells, such as hemopoietic cells, and lower than that in mature parenchymal cells, such as hepatocytes. Furthermore, the progeny of the isolated progenitors can express alpha-fetoprotein and / or albumin and / or CK19. The hepatic progenitors, so isolated, can grow clonally, that is an entire population of progeny can be derived from one cell. The clones of progenitors have a growth pattern in culture of piled-up aggregates or clusters. These methods of isolating the hepatic progenitors are applicable to any vertebrates including human. The hepatic progenitor cell population is expected to be useful for cell therapies, for bioartificial livers, for gene therapies, for vaccine development, and for myriad toxicological, pharmacological, and pharmaceutical programs and investigations.