73 results about "Phytohemagglutinins" patented technology

The invention discloses a CIK cell, as well as a preparation method and a cell preparation thereof. The method for preparing the CIK cell comprises the following steps: placing a separated mononuclear cell in a culture fluid containing phytohemagglutinin; transplanting the mononuclear cell into a culture flask enveloped by antiCD3 monoclonal antibody and antiCD28 monoclonal antibody after the cell is cultured for 24 to 72 hours; and adding a culture fluid containing IL-1alpha and IL-2 into the culture flask to keep on culturing for 5 to 15 days, and separating the cells into different flasks to be cultured every 2 to 3 days. The CIK cell prepared by the method has the characteristics of obvious improved cell proliferation, great increase of CD8 cell proportion, wide antineoplastic spectrum and strengthened antineoplastic activity. The CIK cell and the cell preparation can effectively prevent the metastasis and the recrudescence for of postoperative patients with tumor, can be combined with chemicotherapy to effectively reduce toxic and side effects of the chemicotherapy, strengthens the survivability tolerance of the patients to improve healing efficacy, and can obviously prolong lifecycle to of end-stage patients so as to improve the life living quality.

A method for extracting bean phytohemagglutinin belongs to the extraction technical field of useful constituents of plants. The method is a water extraction method in which the extraction is performed at 40-60 DEG C based on the following steps: crushing bean seeds to 40-100 meshes, adding soak solution to extract bean husk at the temperature of 40-60 DEG C for 60-180min and then fish the bean husk, performing rough separation to remove bean dregs, centrifugally separating to obtain supernatant with the bean phytohemagglutinin, further performing separation and purification on the supernatant, condensing or drying the supernatant to obtain the phytohemagglutinin. Ultrasonic wave or microwave can be exerted in the whole or part of the extraction process, or the ultrasonic wave and the microwave are alternately exerted to perform auxiliary extraction. A traditional low-temperature soaking and low-temperature treatment method is completely discarded in the method, and practice proves that the method is a simple and efficient method for extracting the phytohemagglutinin in short time. Related detection proves that the obtained phytohemagglutinin can meet conventional experimental needs and medicinal needs. The method helps overcome the disadvantage of more chemical reagent composition residue and long extraction time of the products obtained by the prior art.

The invention belongs to the technical field of cell culture in vitro, and particularly relates to a preparation method of high-purity, high-multiplication capacity and high-cytotoxin activity CIK (cytokine induced kill) cell. The method comprises the following steps: collecting and separating peripheral blood mononuclear cell of a patient, eliminating CD4+CD25+Treg cell by means of Mini MACS (magnetic active cell sorting) method, and sorting to obtain CD3+, CD4+ and CD8+T cells; and putting the obtained cells into culture solution containing phytohemagglutinin (PHA), so that the PHA concentration in the suspension liquid is 100ng/ml, hatching for 24h under the culture condition of 5% CO2 at 37 DEG C, transferring the hatched suspension liquid into a cell culture bottle coated by CD3 monoclonal antibody (1mug/ml), adding IFN (interferon)-gamma (1000U/ml), adding IL (interleukin)-2(500U/ml) and IL (interleukin)-21(1000U/ml) after 48h, compensating sodium selenite-containing (0.005mg/L)cell culture after four days, and continuously culturing for 7-14 days to obtain the high-purity, high-multiplication capacity and high-cytotoxin activity CIK (cytokine induced kill) cell. The quantity, the activity and the purity of the CIK cell which is prepared by the method and amplified in vitro are improved, so that the antineoplastic function of the CIK is enhanced.

A highly efficient method of making a primary cell derived biologic by purifying mononuclear cells (MNCs) in a automated cell processor to remove contaminating cells by loading leukocytes onto lymphocyte separation medium (LSM) and centrifuging the medium to obtain purified MNCs, storing the MNCs overnight in a closed sterile bag system, stimulating an induction mixture of the MNCs with phytohemagglutinin (PHA) or other mitogen and ciprofloxacin in a scalable cell culture device and producing a primary cell derived biologic from the MNCs, removing the mitogen from the induction mixture by filtering, incubating the induction mixture, clarifying the induction mixture by filtering to obtain a primary cell derived biologic supernatant, and clearing the primary cell derived biologic supernatant from adventitious agents by anion exchange chromatography, filtration. A closed system prevents contamination of the resulting primary cell derived biologic. An automated method of purifying cells. A method of scalably inducing cells.

The invention discloses an engineering bacterium and a construction method for producing alkaline pectinase immobilized nano micro-spheres by the aid of one-step processes, and belongs to the technical field of immobilized enzymes. A recombinant Escherichia coli strain E.coli BL21 lambda (DE3) pABC-PGL constructed by the aid of the gene recombination method is preserved in the General Biology Center of the China Committee for Culture Collection of Microorganisms, and a strain preservation number of the recombinant Escherichia coli strain is CGMCC NO.10911. The engineering bacterium and the construction method for producing the immobilized alkaline pectinase nano micro-spheres on the basis of the Escherichia coli strains have the advantages that alkaline pectinase genes which are currently industrially widely applied are fused at N tail ends of PHA (phytohemagglutinin) synthesis enzyme genes phaC by connecting peptides by the aid of the gene fusion method, recombinant gene segments are transformed into recombinant bacteria to be expressed in an inducible manner, nano micro-sphere complexes can be synthesized in the recombinant bacteria, alkaline pectinase is carried on the surfaces of the nano micro-sphere complexes and can be effectively combined with the surfaces of the nano micro-spheres to form immobilized alkaline pectinase, and alkaline pectinase immobilization production can be implemented at one step.

The invention relates to an induced culture composition capable of promoting proliferation of a spleen-derived CD8 positive T cell and application of the induced culture composition. The induced culture composition comprises a culture medium and inducing factors added in the culture medium, the inducing factors comprises phytohemagglutinin, a CD3 antibody and a CD28 antibody, the final concentration of the phytohemagglutinin is 1 mu g/mL to 10 mu g/mL, the final concentration of the CD3 antibody is 1 mu g/mL to 10 mu g/mL, the final concentration of the CD28 antibody is 1 mu g/mL to 10 mu g/mL. The induced culture composition provided by the invention can promote proliferation of the spleen-derived CD8 positive T cell.

The invention provides a method for enhancing the reproductive capacity of CIK (Cytokine Induced Killer) cells and improving tumor cell killing capacity. The method comprises the following steps of: sterilely collecting and separating the peripheral blood mononuclear cells of a healthy person; carrying out resuspension on the obtained peripheral blood mononuclear cells by using a Q007 culture medium; adding 24-26 ug/ml of PHA-L type phytohemagglutinin; and culturing for at least 15 days in the Q007 culture medium. The method provided by the invention has the beneficial effects that the reproductive capacity of the CIK cells and the cytotoxic effect are enhanced by adding 24-26 ug/ml of the PHA-L type phytohemagglutinin in a CIK cell culturing process, thereby enhancing the kill rate and the cure effect of the tumor cells.

The invention discloses a method for making soybean-containing nutritious steamed bread. The method comprises the following steps: taking a raw material, boiling soybeans, grinding, kneading dough, fermenting, molding, proofing and steaming. The steamed bread obtained by raw soybean milk or raw soybean flour and cereal fermentation has a very strong soybean flavor, while the soybean phytohemagglutinin also affects the yeast fermentation process, so that the steamed bread is not easily accepted due to poor sensory quality. Therefore, the beany flavor and soybean phytohemagglutinin are ruined after the soybeans are boiled, so that the yeast normally ferments; the boiled soybeans are treated by a colloid mill to reach the micron level, so that the taste of the steamed bread cannot be affected, the soybean flavor of the streamed bread is enhanced, and the sensory level and nutritional value of the steamed bread are improved. The soybean component of the soybean-containing nutritious steamed bread can reach 30 to 50 grams, meeting the nutritional requirements of resident dietary guidelines in China; and if eaten for a long term, the steamed bread has a greater role in promoting the health of all residents.

The invention discloses a preparation method of chromosomes of adult epinephelus akaara and belongs to the field of cell biology. The preparation method comprises the following steps: pretreatment of materials, preparation of kidney cell suspension, low-permeation treatment, fixation, dropping and dyeing, wherein the pretreatment of the materials comprises the steps of with an in vivo injection method of phytohemagglutinin (PHA), injecting PHA into the pectoral-fin base part of the experimental adult epinephelus akaara with 15 micrograms in each gram of the adult epinephelus akaara; after 24 hours, injecting colchicine solution with 2 micrograms in each gram of the adult epinephelus akaara; and after 2.5-3 hours, taking out the head kidney. The preparation method disclosed by the invention has the advantages that the method for one-time injection of PHA is adopted, simultaneously, the injection concentration is increased, and good effect can be obtained after 24 hours, so that the time is saved; and due to the characteristics of tissue cells of the epinephelus akaara, complete cells are not easily obtained when the low-permeation treatment is carried out in the prior art, so that the prior art is creatively changed and a clear chromosome image is obtained.

The invention provides a method for synthesizing PHA (Phytohemagglutinin) by using pseudomonas putida KT2442. The method comprises the following steps of: 1, mixing a small quantity of pseudomonas putida KT2442 in an LB (lysogeny broth) culture medium, and culturing to obtain a seed culture solution containing strains; and 2, adding the seed culture solution obtained in the step 1 and a carbon source to the culture medium, culturing, separating, quick-freezing, drying, and refining to obtain a final product PHA. According to the method, the pseudomonas putida KT2442 in culture wastewater is used, the supply of the carbon source is adjusted, and correlated matrix octane acid and non-correlated matrix glucose are respectively added in the culturing process, thus the final product PHA is obtained. The product obtained by using the method has strong decomposability, low glass transition temperature, good flexibility and adhesion and can be applied to the medicine field, and therefore, the production has a bright application prospect.

The invention provides a lymphocyte serum-free medium with a stable pH value. The lymphocyte serum-free medium comprises the following components: a basic medium, 10 mg/L of transferrin, 0.01 mg/L of sodium selenite, 110 mg/L of pyruvic acid, 1.5 mg/L of ethanolamine, 10 mg/L of insulin, 10-30 mg/L of a glutamine substitute, 300 mg/L of lipid-type bovine serum albumin, 0.001 mg/L of copper sulfate, 0.0005 mg/L of ammonium metavanadate, 0.00005 mg/L of manganese chloride, 0.8 mg/L of zinc sulfate, 2 mg/L of vitamin E, 100 mg/L of phytohemagglutinin (PHA) and 10-50 mM of a pH buffer system. The invention further discloses a preparation method and application of the lymphocyte serum-free medium. Through the adoption of the lymphocyte serum-free medium, lymphocyte differentiation can be improved, and a large number of division-phase lymphocytes are generated; moreover, the pH value is stable , so as to facilitate stable storage of the lymphocyte serum-free medium.

The invention relates to a method for extracorporeally preparing a transfer factor against the specificity of bursal disease viruses of chickens. In the invention, splenic organs or peripheral blood of chickens are used as raw materials to make a lymphocyte monolayer, lymphocytes are extracorporeally cultured, and then phytohemagglutinin and bursal disease viruses of chickens are used to culture the lymphocytes through induction so as to extracorporeally produce a large quantity of transfer factors against the specificity of the bursal disease viruses of chickens. The transfer factor against the specificity of bursal disease viruses of chickens extracorporeally prepared by using the method of the invention is used for treating infectious bursal diseases of chickens and can raise the immunity of poultry.

The invention provides use of hAMSCs (Human Amniotic Mesenchymal Stem Cells) in preparing a drug for treating the acute graft-versus-host disease. The hAMSCs can meet basic biological characteristicsof mesenchymal stem cells of adult tissues, also expresses an embryonic stem cell marker, and has multiplication capacity stronger than that of BMSCs (Bone Marrow Mesenchymal Stem Cells). When the hAMSCs and PHA (phytohemagglutinin) activated healthy human PBMCs (Peripheral Blood Mononuclear Cells) are co-cultured, the proportion of Th1 and Tc1 cell subsets is reduced, the proportion of Th2, Tc2 and Treg cell subsets is increased, the secretion level of IL-2 and IFN-gamma is reduced, the IL-10 level is increased, and therefore, the hAMSCs can be used for preparing the drug for treating the acute graft-versus-host disease.

The invention discloses a method for quantitatively detecting the coagulant activity of bean phytohemagglutinin and belongs to the technical field of plant component detection. According to the method, a red cell suspension for detection is subjected to cell quantification; meanwhile, a coagulant activity concentration standard curve is drawn by using the bean phytohemagglutinin with the known concentration as a standard substance, and the coagulant activity concentration of the bean phytohemagglutinin is further calculated, so that the coagulant activity of the bean phytohemagglutinin is quantified.

The invention relates to a recombinant strain of a mink IFN-Gamma gene, which is characterized by being an IFN-Gamma gene sequence which is cloned from peripheral mink bloodlymph cells (PMBC) simulated by phytohemagglutinin (PHA) and being a recombinant plasmid vector containing a nucleotide sequence; i.e., using a plasmid vector pGM-T which is formed by the cutting of a cloning vector by taking an EcoR V enzyme as the cutting point and then adding a 'T' from the '3' end at both sides; and TA cloning is carried out on the nucleotide sequence of a coding Gamma-interferon which is amplified from the peripheral mink bloodlymph cells so as to construct the recombinant plasmid of the nucleotide sequence of the mink Gamma-interferon. In addition, Escherichia coli conversed from the plasmid vector is used as a host. The recombinant strain aims at laying down foundation for researching and developing genetically engineered mink recombinant interferons biological products with anti-viral activity and immunomodulatory activity.

The invention discloses a method for taking glucose as a substrate to continuously produce polyhydroxylbutyrate valerate (PHBV) in one step by using halophilic mixed bacteria, and belongs to the technical field of PHBV production. The method comprises the following steps: collecting bottom mud at an estuary to inoculate into a fermentation tank, and then operating the fermentation tank by adopting an intermittent operation mode; injecting air into the fermentation tank at a constant speed, controlling the dissolved oxygen concentration in a substrate solution at 1-3mg/L, and continuously running for 5-8 hours, wherein the operation conditions of the fermentation tank are as follows: the constant temperature of the fermentation tank is 37+/-0.1 DEG C, and the pH is 6-8, and discharging 75% mixed solution in the fermentation tank, wherein the mass percent of the PHBV can be up to over 30% by continuous repetition; recovering biomass in the discharged mixed solution, and extracting phytohemagglutinin (PHA). The method is simple to achieve, and engineering operation is easy to carry out, and the produced PHA is a PHBV copolymer, and has high decomposition temperature and low melting temperature. By adopting the method, the production cost of the PHA is reduced by 20-30%.

The invention discloses a bacteriostatic and antiviral preparation taking kidney bean phytohemagglutinin as a major ingredient, and belongs to the technical field of medicines. The bacteriostatic andantiviral preparation consists of the following active components according to relative parts by weight: 0.1-1.0 part of the kidney bean phytohemagglutinin, 0.2-2.0 parts of one or two of stachyose and raffinose as well as one or more of the following active components: 0.1-0.5 part of povidone iodine, 0.1-0.3 part of polyhexylene biguanidine and 0.1-0.5 part of chlorhexidine; and in addition, a thickening agent, which is hydroxyethyl cellulose, is required. The bacteriostatic and antiviral preparation provided by the invention is concrete in components, and is capable of achieving quantitative control and guaranteeing the consistency and stability of overall quality; a concrete inhibitory effect can be achieved on bacteria, fungi and viruses; therefore, the bacteriostatic and antiviral preparation is relatively broad in antibacterial and antiviral ranges; and the product (the bacteriostatic and antiviral preparation), which is required to be preserved at low temperature, is long in preservation duration and convenient to use.

The invention relates to a method of extracting phytohemagglutinin and belongs to the technical field of phytohemagglutinin extraction. The phytohemagglutinin can be extracted from beans by soaking in a buffer solution, acid conditioning to settle protein and adding 0.1-0.4% of sodium chloride solution to settle the protein; the extraction method is simple and convenient; and a reagent used in an extraction process is cheap, easy to obtain and harmless to a human body. Extracted PHA (phytohemagglutinin) protein is high in purity and good in activity; and when the extracted PHA protein is applied to a lymphocyte culture medium, blood coagulation or hemolysis can be avoided, and a good effect of stimulating lymphocyte growth is achieved. The extraction method is suitable for large-scale production of the PHA protein.

The invention belongs to the field of pet foods and especially relates to pet edible pea pods and a preparation method thereof. Peas have high medicinal and nutritional values and are rich in the vitamin C and enzymes being capable for decomposing nitrosamine in body, thereby decomposing the nitrosamine, and also contain abscisic acid, gibberellins and phytohemagglutinin and the like substances, which have anti-bacterial and anti-inflammation effect and metabolism improving function. The Peas also contains abundant dietary fibers that can prevent constipation of pets and has a function of relaxing bowels. Fish skin contains abundant proteins and various micro-elements, wherein the proteins mainly are high-molecular collagen and mucopolysaccharides. Chicken meat is delicious and is easy to digest. In the invention, the pea pod product is prepared from the peas, the fish skin and the chicken meat. The pea pod product has novel and attractive appearance and is more attractive to pet dogs, thereby improving appetite. The peas, the fish skin and the chicken meat are organically combined, thereby improving the nutritional value of the pea pod pet food.

The invention provides a culture medium with a stable pH value. The culture medium is composed of a 1640 base culture medium, a 100 mg/L phytohemagglutinin (PHA), a calf serum with the volume percentage of 20% and a 5-50 mM sodium glycerophosphate/4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) dual-buffering system. The sodium glycerophosphate/HEPES dual-buffering system is composed of a phosphate buffer solution, sodium glycerophosphate and 4-HEPES, wherein the phosphate buffer solution contains Na2HPO4 and KH2PO4. The invention also provides a preparation method of the culture medium and an application of the culture medium in lymphocyte culture. The pH value of the culture medium is not affected by gas composition in the environment, and the pH value during the lymphocyte culture also can be stabilized; and long-term preservation of the culture medium is facilitated.

The invention discloses a preparation method of an enhanced CIK cell, and a cell preparation. The method comprises the following steps: separating to obtain a peripheral blood mononuclear cell from peripheral blood collected from a sufferer; putting the peripheral blood mononuclear cells into a blood-free medium to culture, so as to obtain karyocyte of which the concentration is (1-3)*10<6>/ml; adding IFN (interferon)-gamma, phytohemagglutinin and PGE2 (Phenyl Glycidyl Ether), transferring to a culture bottle or a culture bag to culture for 24 hours, and then adding a CD3 monoclonal antibody, IL-2 (interleukin-2), IL-1alpha, IL-4 and GM-CSF (granulocyte-macrophage colony-stimulating factor); continuing to culture for 3-5 hours, and then controlling the cell density at (1-6)*10<6>/ml; and centrifugally colleting to obtain the enhanced CIK cell after continuously culturing for the 14th to 21st days. The enhanced CIK cell preparation comprises the enhanced CIK cell, human serum albumin, IL-2, amino acids, vitamins and inorganic salts. Compared with a preparation in the patient, the CIK preparation has the advantages that the preservation time of the CIK preparation is obviously prolonged, and is prolonged to 36 hours from 6 hours at room temperature. The CIK preparation is higher in reliability in clinical application, and the safety is effectively ensured.

The invention relates to an in-vitro preparation method of transfer factors capable of resisting duck viral hepatitis. In the method, the transfer factors capable of resisting the duck viral hepatitis are prepared through the induction of duck hepatitis virus by utilizing an in-vitro lymphocyte cell culture method; and chicken spleen or peripheral blood is used as a basic raw material to prepare single-layer lymphocyte cells, and then the single-layer lymphocyte cells are induced and cultured through phytohemagglutinin and the duck hepatitis virus, so that a large number of specific transfer factors capable of resisting the duck hepatitis virus are generated in vitro.. The transfer factors can effectively inhibit the virus of duck viral hepatitis, so that the normal bodies cannot be infected with the duck viral hepatitis, and the radical pre-prevention action can be realized. The method provided by the invention has the characteristics of being simple and easy to operate and the like.

The invention relates to the technical field of plant extraction, and discloses a method for extracting L-type and E-type phytohemagglutinins (PHAs). According to the method, crude protein extraction is carried out through the steps of soaking, crushing, centrifuging, ammonium sulfate fractional precipitation and the like on beans, and then L-type and E-type PHAs are separated and purified through hydrophobic chromatography and ion exchange chromatography. The method is simple to operate, and the extracted L-type and E-type PHAs are high in purity and suitable for large-scale production of the L-type and E-type PHAs.