33results about How to "Get a lot" patented technology

The invention discloses a method for separating the tissue cell of a fetal appendage and constructing a cell bank by means of a tissue homogenate method, so that the process for constructing the cell blank is simple in steps, time-saving, and capable of achieving easy quality control and standardization. The method disclosed by the invention comprises the following steps of (1) cleaning the tissue of the fetal appendage; (2) carrying out homogenate pretreatment on the tissue; (3) carrying out homogenate treatment on the tissue to separate a cell (cell cluster); (4) filtering the tissue and collecting the cell (cell cluster) after the homogenate; (5) detecting the cell (cell cluster); (6) cryopreserving the separated cell (cell cluster) and constructing the cell bank. Compared with the method in the prior art, the method disclosed by the invention has the advantages of simple operation, short time consumption, no introduction of an exogenous reagent, easiness for standardization, easy quality control, great cell gain and the like, and is of great significance in improving the construction efficiency of the fetal appendage cell bank and obtaining the stem cell which is high in quality and applied to treatment.

The invention discloses a gene engineering preparation and mass spectrometry identification method for eriocheir sinensis crustacean hyperglycemic hormone (Ers-CHH). Energy supplies of various tissues and organs are maintained by CHH through regulating a blood sugar level of a crustacean, and researches on family nerve polypeptide hormones can provide helps for revelation of crustacean metabolism, growth and reproduction regulation mechanisms. In previous studies, an eriocheir sinensis crustacean hyperglycemic hormone gene (Ers-CHH, GenBank registration number: JX485664) is cloned. According to the present invention, a gene engineering method is adopted to successfully obtain recombinant Ers-CHH, wherein a molecular weight is about 9.4 kD, a yield after purification renaturation is 0.4 g / L culture medium, and mass spectrometry identification results verify correctness of the recombinant protein. Due to low CHH content in the crustacean, direct purification enrichment on the CHH is virtually impossible, and a large number of the rare hormone proteins are rapidly obtained by using the gene engineering method in the present invention so as to establish a foundation for subsequent function regulation mechanism research development and application in aquaculture production practice.

The invention provides an application of 8-hydroxydeoxyguanosine as a urine marker. The molecular formula of the 8-hydroxydeoxyguanosine is C10H13O5N5. In detection, collection of a sample is non-invasive, and the sample is easy to obtain and is large in acquisition quantity. Compared with FOBT test, the method is more acceptable for detectors and can reduce mental burden of work staff, and is more acceptable for common people. By means of optimized off-line solid-phase extraction technology with ultrahigh performance liquid chromatography-tandem mass spectrometry, the concentration of a DNA oxidative damage marker, 8-hydroxydeoxyguanosine, in urine of a to-be-tested subject can be accurately quantified, so that disease risk analysis to the subject can be carried out through an established colorectal cancer incidence prediction model. The urine marker, when being used for screening the colorectal cancer, is non-invasive, quick and high-flux. The detection method not only is suitable for early stage detection of the colorectal cancer but also is suitable for detection of metastasis or non-metastasis of the colorectal cancer. The application has excellent prospect.

The invention belongs to the technical field of biology and particularly relates to a method for obtaining a large number of regenerated bambusa ventricosa mcclure by tissue culture, comprising the following steps: sterilizing young bambusa ventricosa mcclure branch buds as explants, forming calluses on an improved inducing culturing medium, inducing to generate embryogenic calluses, inoculating the embryogenic calluses on an improved differentiating culturing medium, inducing the embryogenic calluses to differentiate and germinate, inducing the embryogenic calluses to differentiate and root on a rooting culturing medium to form complete plants, and transplanting the plants into flower pots to finish the whole regeneration process of the bambusa ventricosa mcclure. The method can be used for obtaining the bambusa ventricosa mcclure at high frequency and is suitable for the genetic improvement of the bambusa ventricosa mcclure.

The invention discloses a novel sedimentation pool for a sewage plant. The novel sedimentation pool comprises a pool body, a water inlet pipe, a water distribution unit, a biological filling materialunit, a mud scraping device, a gas recovery device and a water outlet pipe, wherein the water distribution unit, the biological filling material unit, the mud scraping device, the gas recovery deviceand the water outlet pipe are arranged in the pool body; the mud scraping device comprises a mud scraping rod arranged at the bottom surface of the pool body, a driving rod fixedly connected with themud scraping rod, a rotating driving element used for driving the driving rod to rotate and a second driving element used for driving the driving rod to vertically act; the water distribution unit comprises a center pipe, a plurality of water distribution pipes and spray heads arranged on the water distribution pipe; the biological filling material unit comprises a first filling material assembly;the first filling material assembly comprises a fixing sleeve, a plurality of filling material frames and a biological filling material; the filling material frames are distributed at intervals in the peripheral direction of the fixing sleeve; the biological filling material is arranged on the filling material frame. The sewage is in sufficient contact with the biological filling; the biologicalfilling has a good anaerobic digestion treatment effect on the sewage; the sewage deposition quantity is reduced; the frequent mud scraping operation is not needed; the energy is saved; the environment is protected; the abrasion of the mud scraping component is small; the service life is long.

The invention discloses micro-carbon high-chromium high-boron wear-resistant steel and a preparation method thereof. The prepared wear-resistant steel comprises the following components of, in percentage by weight, 0.10-0.25% of carbon, 1.50-3.00% of silicon, 0.80-3.00% of manganese, less than 0.04% of phosphorus, less than 0.04% of sulfur, 9.00-15.00% of chromium, 0.30-3.00% of nickel, 0.10-1.00%of molybdenum, 0.10-0.50% of vanadium, 0-1.50% of tungsten, 0.01-0.10% of titanium, 0.40-0.80% of boron, 0.10-0.40% of copper, 0.10-0.30% of aluminum, 0.02-0.25% of niobium, 0.02-0.10% of zirconium,0.10-0.50% of cerium, 0.01-0.15% of tin, 0.001-0.003% of magnesium, 0.001-0.15% of calcium, 0.003-0.025% of nitrogen, 0.001-0.005% of rhenium, and the balance iron. The steel not only has the high hardness, also has the characteristics of certain toughness and high wear resistance at the same time.

The invention discloses a method for generating an alfalfa genome edited homozygous plant, and belongs to the technical field of plant genetic engineering. The method mainly comprises: establishing an alfalfa hairy root system; converting alfalfa on a CRISPR / Cas9 over-expression vector, and obtaining resistant hairy roots; quickly filtering a homozygous hairy root system having an alfalfa genome edited by a fixed point; establishing a hairy root regeneration system, and obtaining a genome edited homozygous alfalfa plant. The method is mainly used for solving the problems of large workload and long period for obtaining a CRISPR / Cas9 genome edited homozygous plant. By means of the method of the invention, an alfalfa genome edited homozygous hairy root system can be obtained within 1-2 months; all alfalfa plants obtained by regenerating the homozygous hairy root system are homozygous edited plants.

The invention discloses a method for quickly breeding cymbidium hybridium by use of root inducing protocorm. The improved MS culture medium containing special components (6-BA, PIC and casein hydrolysate) is used for inducing protocorm; for the root explant of cymbidium hybridium, the protocorm is induced by the root of cymbidium hybridium through a special cultivation method, wherein the induction rate reaches 100%; and after 45 days of cultivation, 50 protocorms of each root explant are obtained, the survival rate of the protocorm is 95%, and the plant grows well and directly roots.

The invention discloses a co-immune vaccine for prevention and / or treatment of diseases caused by respiratory syncytial virus infection. The vaccine is composed of a DNA vaccine and a subunit vaccine. The DNA vaccine is a recombinant expression vector containing an encoding gene of a protein as shown in a sequence 2 in a sequence table. The subunit vaccine is a protein as shown in the sequence 2 in the sequence table. Experimental results show that the co-immune vaccine provided by the present invention can enhance the humoral immune response of immune animals, also inhibit too strong cellular immune response, effectively inhibit inflammation, protect lung tissues of mammalians and prevent function from destruction of RSV infection.

The invention discloses a method for cloning male genomes of lepidopterous insects, which comprises the following steps: (1) radiating female pupae of receptor insects in 2d-3d before eclosion to destroy oocyte nuclei; (2) extracting semen of male moths of donor insects; (3) carrying out artificial insemination on virgin moths of the receptor insects; (4) collecting ova propagated by female moths after artificial insemination treatment; (5) carrying out androgenesis treatment on the propagated ova obtained in the step (4) to obtain androgenesis ova; and (6) screening and treating the androgenesis insect ova obtained in the step (5), and carrying out hatching treatment to obtain G1-generation male genome clones of the donor insects. The method of the invention can avoid injury of nucleus transplantation required for a nucleus transplantation cloning technology and an artificial reproduction method, fusion treatment of sperm nuclei and ovoplasm and genome activation stimulation, has low aberration rate of cloned posterity, can obtain individual posterity, tissues and cells which are highly consistent with sperm donor insects, and can obtain high-quality materials for evaluating influence of ovoplasm to cloned animals.

The invention discloses a preparation method of a high-capacity serum antibody. The preparation method comprises the following steps of firstly, collecting immunized fresh blood, collecting blood plasma, and cryopreserving for later use; then, filtering the collected blood plasma, and collecting filtrate to obtain a primarily-filtered serum product; and adding thrombin into the prepared primarily-filtered serum product, uniformly stirring, warmly applying at the temperature of 20-37 DEG C for 0.5-4 hours, filtering by using filter paper with the aperture of 0.22-0.88mu m, and collecting filtrate to obtain the serum antibody. The serum antibody obtained by using the method disclosed by the invention is high in content and good in quality and stability; and the preparation method is suitable for preparing the high-capacity serum antibody and relatively good in market prospect.

The invention discloses a passiflora foetida tissue culture method. The method comprises the following steps: with leaves of passiflora foetida as explants, disinfecting to obtain explants with a lowpollution rate and less damage, inducing the successfully disinfected explants in induction media of different formulations to obtain somatic embryos, adventitious buds and other different products, transferring the somatic embryos into somatic embryo germination media for continuously culturing to obtain germinated somatic embryos, and carrying out plant regeneration culture, transplantation andthe like to obtain passiflora foetida plants growing well. The method has the advantages of convenient material selection, high material utilization rate, large quantity and simple operation, is suitable for factory production, establishes a tissue-culture rapid propagation system with the passiflora foetida leaves as the explants, and provides a seedling technology for rapid propagation of passiflora foetida.

The invention provides a mechanical ventilation man-machine asynchronous data acquisition method, a mechanical ventilation man-machine asynchronous data detection method and mechanical ventilation man-machine asynchronous data detection equipment. The mechanical ventilation man-machine asynchronous data acquisition method comprises the following steps: starting and connecting a respirator and a simulated lung; setting a simulated breathing mode of the simulated lung through simulated lung control software according to a preset type of built-in case; adjusting the mechanical ventilation mode of the breathing machine to be not matched with the simulated breathing mode of the simulated lung, so that a man-machine asynchronous phenomenon is generated between the breathing machine and the simulated lung; and when the man-machine asynchronous phenomenon is stable, exporting related man-machine asynchronous data through the simulated lung control software. Through the above mode, the asynchronous data acquisition method provided by the invention can quickly and efficiently acquire different man-machine asynchronous data in a large amount and various types through the cooperation of the breathing machine and the simulated lung, and provides an important preposed basis for related teaching, research and application.

The invention relates to a miraculin recombinant protein and an expression and purification method thereof. The carboxyl terminal of the miraculin recombinant protein expression vector used in the method is provided with a FLAG label, and Escherichia coli Origami (DE3) is adopted, so that a large amount of miraculin recombinant protein can be rapidly obtained, and a new method is provided for solving the problem of miraculin source and yield limitation, meanwhile, rich raw materials are provided for developing a new sweetening agent and researching a taste changing mechanism of the miraculin.

The invention belongs to the technical field of biology and particularly relates to a method for obtaining a large number of regenerated bambusa ventricosa mcclure by tissue culture, comprising the following steps: sterilizing young bambusa ventricosa mcclure branch buds as explants, forming calluses on an improved inducing culturing medium, inducing to generate embryogenic calluses, inoculating the embryogenic calluses on an improved differentiating culturing medium, inducing the embryogenic calluses to differentiate and germinate, inducing the embryogenic calluses to differentiate and root on a rooting culturing medium to form complete plants, and transplanting the plants into flower pots to finish the whole regeneration process of the bambusa ventricosa mcclure. The method can be used for obtaining the bambusa ventricosa mcclure at high frequency and is suitable for the genetic improvement of the bambusa ventricosa mcclure.

The invention mainly relates to a clone, an expression and a protein purification method of an OPN associated protein pro-MMP-9 zymogen. The method includes the steps of recombination of the structure of a cloning vector TA-pro-MMP-9, recombination of the structure of an expression plasmid pET-28a(+)-pro-MMP-9 and induced expression and identification of pro-MMP-9 protein. A prokaryote genetic expression system is built, so that a lot of pro-MMP-9 can be expressed, high-purity protein purification procedures are built, and pro-MMP-9 protein which is large in amount, high in purity and simple and feasible in preparing method is obtained.

An ethylene-modified PVA-based polymer film comprising an ethylene-modified PVA-based polymer having good stretchability and productivity and having an ethylene unit content of 1 to 4 mol%, the film satisfying the following formulas (I) and (II) ). Δn(MD)Ave‑0.1×10‑3≤Δn(TD)Ave≤Δn(MD)Ave+0.25×10‑3 (I) Δn(TD)Ave≤2.5×10‑3 (II) [in the above formula , Δn(MD)Ave represents the value obtained by averaging the birefringence in the direction of mechanical flow of the ethylene-modified PVA-based polymer film in the thickness direction of the film, Δn(TD)Ave represents the value of the ethylene-modified PVA-based polymer The birefringence in the width direction of the film is averaged in the thickness direction of the film.

The invention discloses a gene engineering preparation and mass spectrometry identification method for eriocheir sinensis crustacean hyperglycemic hormone (Ers-CHH). Energy supplies of various tissues and organs are maintained by CHH through regulating a blood sugar level of a crustacean, and researches on family nerve polypeptide hormones can provide helps for revelation of crustacean metabolism, growth and reproduction regulation mechanisms. In previous studies, an eriocheir sinensis crustacean hyperglycemic hormone gene (Ers-CHH, GenBank registration number: JX485664) is cloned. According to the present invention, a gene engineering method is adopted to successfully obtain recombinant Ers-CHH, wherein a molecular weight is about 9.4 kD, a yield after purification renaturation is 0.4 g / L culture medium, and mass spectrometry identification results verify correctness of the recombinant protein. Due to low CHH content in the crustacean, direct purification enrichment on the CHH is virtually impossible, and a large number of the rare hormone proteins are rapidly obtained by using the gene engineering method in the present invention so as to establish a foundation for subsequent function regulation mechanism research development and application in aquaculture production practice.

The invention discloses a method for separating the tissue cell of a fetal appendage and constructing a cell bank by means of a tissue homogenate method, so that the process for constructing the cell blank is simple in steps, time-saving, and capable of achieving easy quality control and standardization. The method disclosed by the invention comprises the following steps of (1) cleaning the tissue of the fetal appendage; (2) carrying out homogenate pretreatment on the tissue; (3) carrying out homogenate treatment on the tissue to separate a cell (cell cluster); (4) filtering the tissue and collecting the cell (cell cluster) after the homogenate; (5) detecting the cell (cell cluster); (6) cryopreserving the separated cell (cell cluster) and constructing the cell bank. Compared with the method in the prior art, the method disclosed by the invention has the advantages of simple operation, short time consumption, no introduction of an exogenous reagent, easiness for standardization, easy quality control, great cell gain and the like, and is of great significance in improving the construction efficiency of the fetal appendage cell bank and obtaining the stem cell which is high in quality and applied to treatment.

The invention discloses a primer system. The primer system comprises a primer 1, a primer 2, a primer 3 and a primer 4, wherein the nucleotide sequence of the primer 1 is shown in SEQ ID No:1, the nucleotide sequence of the primer 2 is shown in SEQ ID No:2, the nucleotide sequence of the primer 3 is shown in SEQ ID No:3, and the nucleotide sequence of the primer 4 is shown in SEQ ID No:4. The invention also discloses an expression vector and a transgenic cell and applications of the expression and transgenic cell. Through the technical scheme above, the heterologous in vitro expression of krait antibacterial peptides CBF is realized for the first time, and a purpose of rapidly obtaining large quantities of CBF in a low-cost mode is successfully achieved, thereby providing a theoretical basis for the large-scale development and utilization of CBF.

The invention discloses a young pig islet cell culture medium and an application method thereof. The culture medium is added into a basic culture medium (M199): 1-50 mM of glutathione, 1-50 mM of glutamine, ITS 0.01-1 % of ITS, 1-50 mM of nicotinamide, 0.1-20 micro-M of water-soluble vitamin E, 1-100 U / L of heparin, 0.01-0.5 mM of 4-(2-aminoethyl) benzenesulfonyl fluoride hydrochloride and 0.01-0.1 mM of exenatide. Compared with the common media (1640, F10 and EGM2), the viability and maturation rate of islet cells in young pigs can be significantly improved.

The invention discloses a dragon fruit tissue culture method using blades to induce adventitious buds. The dragon fruit blades are used as explants and are disinfected to obtain explants with extremely low pollution rate and less damage, the successfully-disinfected explants induce adventitious buds in an adventitious bud induction medium, and then dragon fruit plants growing well are obtained through rooting and proliferation, transplanting and other processes. The dragon fruit tissue culture method has the characteristics of short culture period, high speed and quick emergence, has the advantages of high material availability, high material utilization rate, short breeding period and simple operation, is suitable for factory production, establishes a tissue culture rapid propagation system with dragon fruit blades as explants, and provides seedling technology for rapid propagation of the dragon fruit.

A PVA film that satisfies expressions (I) and (II), which has satisfactory stretchability and from which a polarizing film can be efficiently produced; and a process for film production which comprises (a) ejecting a PVA-containing film formation dope in a film form onto the first drying roll of a film formation device equipped with a plurality of drying rolls, partly drying the web, and then successively drying the web with the subsequent drying rolls to produce a film, wherein (b) the ratio of the peripheral speed (ST) of the drying roll on which the volatile content of the PVA film has fallen to 13 mass% to the peripheral speed (S1) of the first drying roll, ST / S1, is regulated to 0.990-1.050, (c) the ratio of the peripheral speed (SL) of the last drying roll to the peripheral speed ST, SL / ST, is regulated to 0.960-0.980, and (d) the SL / S1 ratio is regulated to 0.970-1.010. ?n(MD)Ave-0.1×10-3=?n(TD)Ave=?n(MD)Ave+0.25×10-3 (I) ?n(TD)Ave=2.5×10-3 (II) [In the expressions, ?n(MD)Ave indicates the value obtained by averaging the machine-direction birefringence values of the PVA film with respect to the film thickness direction, and ?n(TD)Ave indicates the value obtained by averaging the transverse-direction birefringence values of the PVA film with respect to the film thickness direction.]

The invention belongs to the technical field of chemical medicine preparation, and particularly relates to a latamoxef hydroxyl impurity preparation method which comprises comprises the steps: mixing a latamoxef sodium aqueous solution and a sodium hydroxide aqueous solution, and carrying out a heat preservation reaction to obtain a; and then separating the a by using macroporous resin, and freeze-drying. The preparation method has the advantages of being simple and easy to operate, high in purity and large in sample obtaining amount. The prepared latamoxef hydroxyl impurity is convenient for quality research of latamoxef sodium, especially provides a large batch of impurity reference substances with uniform quality for quantifying the impurity content in latamoxef sodium by adopting an external standard method, is beneficial to improving the overall quality level of latamoxef sodium, and lays a foundation for the variety to reach the quality level of a reference preparation.