1287 results about "Co-Culture" patented technology

The invention discloses a vascularized immune co-culture organoid chip for simulating a tumor microenvironment. The vascularized immune co-culture organoid chip comprises a chip main body and a bottom plate, a flow channel and a liquid storage tank are arranged in the chip main body; the runners comprise a first runner, a second runner, a third runner and a fourth runner; a compartment is arranged in the middle of the third flow channel; the first flow channel is connected with a first liquid storage tank which is used for injecting a first hydrogel suspension obtained by mixing fibroblasts and hydrogel into the first flow channel; the second flow channel is connected with a second liquid storage tank for injecting a culture solution into the second flow channel; a compartment is arranged in the middle of the third flow channel; the third flow channel is connected with a third liquid storage tank communicated with the compartment, and the compartment is connected with a compartment liquid storage tank; the compartment liquid storage tank is used for injecting a second hydrogel suspension obtained by mixing tumor organs and hydrogel into the compartment, and the third liquid storage tank is used for injecting hydrogel into the third flow channel; and the fourth flow channel is connected with a fourth liquid storage tank for injecting vascular endothelial cell and immune cell suspension into the fourth flow channel. The invention further discloses a vascularized immune co-culture organoid culture method for simulating the tumor microenvironment.

A method and system to make a modular microfluidic device in a miniaturized form for cell culture, co-culture, and organ-on-a-microfluidic device as in-vitro model of human organs within compartmentalized 3D structures is described. The modular microfluidic device enables co-culturing of heterogeneous cell assemblies that are adjacent or sequentially connected to promote enhanced cell-cell interaction. In vivo-like flow into and / or from one cell growth chamber to adjacent cell growth chamber is maintained by micro-engineered porous barriers i.e., porous walls and membranes. These porous barriers provide a tool for mimicking the paracrine exchange between cells in the human body. The system comprises of a set of microelectrodes for real-time and long-term monitoring and quantitative measurement of the impact of stimuli on the epithelial tissue permeability. The modular microfluidic system provides a platform technology for bio-fabrication of variety of miniaturized in-vitro models for mimicking the structure and function of single and multi-organ models.

The invention relates to the technical field of biological information, in particular to an immunoreaction evaluation method based on tumor neoantigen activity sorting. The method comprises the following steps: obtaining genome data of tumor and normal tissues through whole exon sequencing, extracting multi-dimensional features of candidate somatic mutation, and screening by combining a Gaussian mixture model and a Transform model to obtain a high-confidence mutation genome set; predicting the HLA genotype of a patient based on sequencing data, translating and mutating into a peptide fragment, predicting the binding affinity of the peptide fragment and an MHC molecule by using an XGBoost model, and calculating a new antigen activity score sequence in combination with various parameters; and finally, synthesizing a new antigen peptide fragment according to a sorting result, carrying out in-vitro co-culture to detect an IFN-gamma secretion result, and dynamically optimizing a characteristic combination coefficient through a PPO algorithm to realize intelligent iterative updating, so that the accuracy of new antigen screening and the immunoreaction prediction capability are remarkably improved.

The invention provides a neuromuscular multi-element coupled organoid micro-fluidic chip, a device and a method. Relates to the technical field of organoid micro-fluidic chips. A first culture chamber, a second culture chamber and a third culture chamber are arranged in a culture layer of the chip; the first culture room is used for culturing nerve cells; the second culture room is used for culturing muscle cells; a microfluidic channel for communicating the first culture chamber with the second culture chamber is arranged between the first culture chamber and the second culture chamber; the third culture room is arranged on the micro-fluidic channel and is used for culturing a neuromuscular joint formed by convergence of axons of nerve cells and muscle cells; a microelectrode array plate is arranged in the circuit layer and is used for performing electrical stimulation and electrophysiological function detection on neuromuscular cells. Through the chip, co-culture of in-vitro nerves and muscle organs can be realized, a complex interaction relationship in a neuromuscular system can be accurately simulated, and the chip can also be used for constructing various neuromuscular related disease models.

The invention relates to the technical field of biological tissue engineering, and discloses a nerve-blood brain barrier co-culture model, which comprises a culture chip, and nerves and blood brain barriers respectively constructed in upper culture micropores and lower culture micropores of the culture chip. Wherein a semi-permeable membrane is arranged between the lower-layer culture micropores and the upper-layer culture micropores of the culture chip; the nerves are obtained by inoculating neurons / brain organs obtained by stem cell differentiation into the upper culture micropores for co-culture, and the blood brain barrier is obtained by inoculating brain microvascular endothelial cells obtained by stem cell differentiation into one side of the semipermeable membrane in the lower culture micropores for co-culture. The co-culture model is higher in biomimetic property, and can better simulate the structure and function of the blood brain barrier, for example, the co-culture model has a barrier function closer to the physiological level, the selective permeability of a small molecule drug is matched with the in-vivo permeation condition, and the co-culture model has the function of a small molecule related transporter. The invention further discloses a construction method and application.

The invention belongs to the technical field of tissue engineering, and particularly discloses a preparation method of a 3D full-thickness skin model. The preparation method provided by the invention comprises the following steps: constructing a corium layer, constructing an inner epidermal layer through first epidermal inoculation, constructing an epidermal layer through second epidermal inoculation and obtaining a complete 3D full-thickness skin model through co-culture. The 3D full-thickness skin model obtained through the preparation method is distinct in gradation, uniform in thickness, free of shrinkage and stable in quality, the epidermal layer and the corium layer are uniformly fused, and the 3D full-thickness skin model is suitable for skin function research and sample evaluation. Meanwhile, the preparation method provided by the invention is simple in process and convenient to operate, does not need additional reagents or anti-shrinkage tools, can meet the requirements of large-scale industrial production, and has good production feasibility and market application prospects.

The invention discloses a preparation method of NK (Natural Killer) cells. The method comprises the following steps: (1) separating and purifying the NK cells; (2) primarily amplifying the NK cells in a low-oxygen environment; (3) co-culturing the NK cells; and (4) continuously amplifying and harvesting the NK cells. The NK cell prepared by the method shows relatively high cell purity and activity, and shows a remarkable anti-tumor effect in a malignant tumor treatment mouse model. The method has the advantages of high efficiency and reliability, and provides good reference for research and clinical application of the NK cells.

The invention belongs to the technical field of organoid culture, and particularly relates to a co-differentiated vascularized liver organoid and a construction method thereof. Aiming at the problems that the existing liver organ vascularization mostly adopts a co-culture mode, the operation process is complicated, the cell source heterogeneity is relatively high, the randomness is strong, the uniformity is poor and the like, the invention provides the construction method of the co-differentiated vascularized liver organ. The preparation method comprises the following steps: sequentially inducing and culturing directional endoderm cells, posterior preintestinal cells and hepatic endoderm cells, adding an S1 culture medium containing VEGF to induce and culture for 1-3 days, then adding an S2 culture medium containing VEGF, BMP4 and FGF2 to induce and culture for 3-5 days, and then adding an S3 culture medium containing VEGF to induce and culture for 14-20 days to obtain the co-differentiated vascularized liver organ. According to the method disclosed by the invention, the operation process is simplified, the experiment efficiency is improved, the liver organ which is rich in blood vessel network, high in maturity and of a blood vessel-bile duct structure is obtained, and a foundation is laid for large-scale production and clinical application.

The invention relates to a medical simulation experiment method and device for acting apoptosis vesicles on primary cartilage cells based on cell line culture, and the method mainly comprises the following steps: S1, co-culture system construction, S2, multi-dimensional observation and analysis, and S3, operation risk assessment. In the step S1, primary cartilage cells, primary immune cells and cell line culture apoptosis vesicles are co-cultured; s2, carrying out multi-dimensional observation and analysis on the influence of apoptosis vesicles on cells; and S3, mainly constructing a multi-dimensional risk scoring system, and carrying out risk grade division by applying a biological network model. The invention aims at simulating and evaluating the influence of cell line culture apoptosis vesicles on primary cartilage cells in vitro and the interaction between the cell line culture apoptosis vesicles and primary immune cells, so that an experimental basis is provided for subsequent clinical application.

The invention provides a bone organ model as well as a construction method and application thereof, and the method comprises the following steps: arranging a biological material with a micropore array in a cell culture plate to obtain a bone organ chip; suspending the initial stem cells in a culture medium, and promoting the cells to form a spherical structure through cell aggregation or by using a non-adhesive culture surface, so as to obtain cell spheres; and culturing the cell spheres in the bone organ chip by adopting the culture medium for constructing the bone organ model to obtain the bone organ model. The skeletal organ is successfully constructed through in-vitro self-organized 3D co-culture initial stem cells including embryonic stem cells, induced pluripotent stem cells, bone marrow mesenchymal stem cells, osteoblasts, bone cells, all levels of osteogenic precursor cells, osteogenic-like cells and MC3T3-E1 cells, and the skeletal organ can become an important tool for researching and treating bone related diseases.

The invention belongs to the technical field of medicines, and particularly discloses application of moss phenol glucoside in preparation of medicines for preventing and / or treating tumors. According to the method, activated CD8 + T cells, MC38-OVA cells and traditional Chinese medicine monomeric compounds are co-cultured, the traditional Chinese medicine monomeric compounds capable of effectively promoting the anti-tumor activity of the CD8 + T cells are selected by utilizing a high-content screening technology, and finally, the experiment is carried out by taking the moss phenol glucoside as a research object. In-vitro experiment results show that the moss phenol glucoside can significantly promote T cell activation, promote release of cell factors such as IFN-gamma and TNF-alpha, and enhance the killing effect of T cells on tumors. A mouse in-vivo tumor-bearing experiment result shows that the moss phenol glucoside can promote the anti-tumor activity of T cells so as to inhibit tumor growth, has no obvious toxic or side effect, can be used for preparing the medicine for preventing and / or treating tumors, and has a potential clinical application prospect and a relatively high economic value.

The invention provides a preparation method of a cell culture three-dimensional scaffold and cell culture meat, which comprises the following steps: firstly, preparing a gelatin-carrageenan mixed solution and a gel precursor solution of transglutaminase, dividing into two parts, incubating at constant temperature to obtain two parts of composite hydrogel, randomly freeze-drying one part to obtain aerogel, and freeze-drying the aerogel to obtain the cell culture three-dimensional scaffold. Stacking on the other part and incubating at constant temperature again to obtain the prefabricated double-layer gel. The prefabricated double-layer gel is directionally frozen, ice crystals grow upwards from the bottom, freezing is stopped after the temperature is reduced to the set temperature, and the cell culture three-dimensional scaffold is obtained through drying. The stent adopts an ordered-disordered partition design, parallel micro-channels are formed in the lower layer by using a directional ice template method, and muscle fiber arrangement is simulated; a porous spongy structure is constructed on the upper layer by using a random ice template method, and is adapted to adipocyte proliferation. Two layers of interfaces are interpenetrated and communicated, natural mechanical gradients are combined, natural tissue characteristics are re-engraved, the traditional limitation is broken through, the cell co-culture efficiency and the cultured meat quality are improved, and industrialization is promoted.

The present invention belongs to the field of organoid chips. Provided is a multiple-organoid co-culture micro-fluidic chip, comprising organoid micro-fluidic chip cells, each organoid micro-fluidic chip cell being divided into an upper layer and a lower layer which are bonded to each other, the upper layer being sequentially provided with an organoid culture medium perfusion channel, a first channel barrier, an organoid culture chamber, a second channel barrier and a vascular endothelial culture channel, and the lower layer being provided with a micro-perfusion network channel. The chip ensures specific nutrition and microenvironment for each organoid and helps to maintain specific phenotypes for same. In addition, the chip can simulate in-vitro blood flowing, so as to perform signal communication and interaction between organoids, thereby fully simulating the in-vivo environment and interaction for multiple-organ diseases.

The present invention relates to compositions comprising a microbial consortium for treating or preventing medical disorders and pathologies through microbiome modulation, including medical disorders associated with a dysbiosis or a loss of gut microbial function.

The invention provides siRNA for reducing IFNGR1 and application of the siRNA, and relates to the technical field of biology, the siRNA comprises at least one of IFNGR1-si-1, IFNGR1-si-2 or IFNGR1-si-3, and experiments prove that the knock-down efficiency of the IFNGR1-si-1, the IFNGR1-si-2 and the IFNGR1-si-3 on IFNGR1 genes in mesenchymal stem cells reaches 48% or above. And quantitative expression of the mesenchymal stem cell IFNGR1 can be realized by quantitatively transferring siRNA. A fitting formula is determined by establishing a corresponding relation between the IFNGR1 expression quantity and the total lymphocyte proliferation inhibition rate, and the total lymphocyte proliferation inhibition rate can be calculated by detecting the IFNGR1 expression quantity of the mesenchymal stem cells to be evaluated. Compared with the prior art, the MSC does not need to be evaluated to be co-cultured with the PBMC every time, and a large number of PBMCs are used.

The present invention relates to an intestinal epithelium model accurately reproducing the pathophysiological mechanisms observed in vivo in the context an inflammatory condition of the intestinal barrier. The model according to the present invention comprises two compartments separated by a semi-permeable membrane. The first compartment, corresponding to the apical pole of the intestinal epithelium, comprises a coculture of Caco-2 cells differentiated into enterocytes and HT29-MTX cells differentiated into goblet cells. The second compartment, corresponding to the basolateral pole of the intestinal epithelium, comprises a culture of THP-1 monocytic cells differentiated into macrophages. The model according to the present invention is characterized in that the cells contained in the first compartment produce interleukin 6 (IL-6) at a concentration of more than 100 pg / ml and interleukin 8 (IL-8) at a concentration of more than 150 pg / ml, and the cells contained in the second compartment produce tumour necrosis factor α (TNF-α) at a concentration of more than 40 pg / ml and interleukin 1β (IL-1β) at a concentration of more than 90 pg / ml. The present invention also relates to the method for obtaining this model and also to a method for selecting candidate compounds for the treatment of inflammatory bowel diseases by verifying their ability to stop the inflammation as soon as the permeability is increased, or verifying the restoration of the permeability and of the mucus layer.

The invention discloses a method for establishing a co-culture system of porcine skeletal muscle satellite cells (PSCs) and porcine kidney fibroblasts (PKFs). The method comprises the following steps of: performing in-vitro induction on the PKFs to transdifferentiate into adipocytes, optimizing a process of the method, establishing an optimal PSCs and PKFs co-culture induced differentiation method, and selecting a co-culture proportion. According to the method for in-vitro induction of PKFs transdifferentiation and the co-culture system of PKFs and PSCs established by the invention, the PKFs can replace porcine precursor adipocytes (ADSCs) to serve as a source of seed cells in artificial meat fat, and the period of adipogenesis induced differentiation is shortened; the established PKFs and PSCs co-culture system provides a new two-dimensional model for the artificial snowflake meat; a powerful in-vitro model is provided for deep understanding of intercellular interaction and endocrine and paracrine functions of the skeletal muscle which is a highly heterogeneous tissue.

A method for enhancing hydrogen uptake by cells of an acetogenic bacterium in a culture is provided. The method comprises incubating the cells in a liquid medium supplemented with an external gas mixture, wherein the external gas mixture comprises H2 and CO2; overexpressing an uptake hydrogenase and an associated electron transfer protein in the cells; increasing uptake of the H2 by the cells; and enhancing uptake of the CO2 by the cells. The culture may be a coculture with additional cells of a non-acetogenic bacterium, which produces natively H2 from catabolism of an organic molecule.

The invention relates to a culture method for activating NKT cells by DC cells in vitro and application of the NKT cells. The preparation method of the NKT cell comprises the following steps: activating a DC cell by using cell inducing factors such as alpha-GalCer, GM-CSF and IL-4, co-culturing the DC cell and a cryopreserved and resuscitated suspension cell left after adherent sorting, and activating and amplifying the NKT cell. The proportion of NKT (CD3 + CD56 +) prepared by the invention is more than 80%, and a small amount of iNKT cells are contained at the same time.

The invention discloses an application of a CD73 + NK cell in preparation of a medicine for preventing or treating Parkinson's disease. The applicant discovers that the CD73 + NK cell can prevent dopaminergic neuron loss and glial inflammation response of the MPP + induced VM cell in the experimental process. However, TGF-beta1 and IL-10 secreted by the CD73 + NK cells do not significantly prevent loss of dopaminergic neurons induced by MPP + in cross-pore co-culture of the CD73 + NK cells and VM neurons. Therefore, the CD73 + NK cells are proved to protect dopaminergic neurons from the influence of MPP + neurotoxicity through an intercellular contact mechanism, namely CD73-SIRPA interaction.

The invention relates to a cell collection method capable of simultaneously collecting three cells in a co-culture model, and belongs to the technical field of biology. The invention provides a cell collection method capable of simultaneously collecting three cells in a co-culture model, and the cell collection method comprises the following steps: after a three-cell co-culture model is constructed, taking out a Transwell chamber, retaining cells in the lower chamber, and collecting the cells in the lower chamber; respectively digesting the cells on the two sides of the Transwell cell membrane by using a trypsin solution with the concentration of 0.5 g / 100mL so as to respectively collect the cells on the two sides of the Transwell cell membrane. According to the cell collection method disclosed by the invention, the three cells in the three-cell co-culture model are simultaneously collected in a manner of digesting the cells on the two sides of the Transwell membrane step by step by using pancreatin, so that not only is the cell and consumable cost saved, but also the experimental synchronism of the three cells is ensured, and convenience is provided for optimizing the experimental process.

The utility model relates to the technical field of cell cultivators, in particular to a pump-free perfusion micro-fluidic chip for multi-cell three-dimensional co-culture, which comprises a liquid storage layer, a liquid inlet and outlet layer, an upper channel layer, a lower channel layer and a packaging layer, realizes a one-way circulation perfusion dynamic co-culture system without a pumping system, reduces the occupied space of equipment, and is convenient to operate. Operation is simple, culture medium consumption is reduced, and cost is reduced; the separation of free NK cells and tumor infiltration NK cells is realized, and conditions are provided for researching molecular biological characteristics of different types of NK cells. The migration and interaction of the tumor spheres and the NK cells are observed in real time at the same time, so that the problem that a microfluid pump integrated by an external system is needed for separating free immune cells and tumor infiltration immune cells by existing equipment is solved.

The invention provides a method for enhancing Treg cell functions and application, and belongs to the technical field of cell engineering. According to the method, hUC-MSCs are irradiated by gamma rays to form feeder layer cells, and then the feeder layer cells and Treg cells are co-cultured to obtain the Treg cells with enhanced cell functions. Through the co-culture of the feeder layer cells and the Treg cells, the inhibition ability of the Treg cells on activation and proliferation of immune cells can be enhanced, and the feeder layer cells and the Treg cells can be used as universal cells in clinical researches of autoimmune diseases, graft versus host diseases and the like.

The invention provides an in-vitro skin blood vessel immune model as well as a preparation method and application thereof. THP-1 human monocyte leukemia cells are induced to be differentiated into M0 type macrophages, and then the M0 type macrophages are respectively induced into M1 type macrophages and / or M2 type macrophages; then co-culturing the M1 type and / or M2 type macrophages and vascular endothelial cells to form a vascular immune model; finally, the 3D skin model is placed on the blood vessel immune model, external stimulation is conducted, and the in-vitro skin blood vessel immune model is constructed. The in-vitro skin blood vessel immune model can be used for repairing skin barriers, inflammation pathways, blood vessel metabolism, the expression level of genes or proteins related to extracellular matrixes, vascular endothelial cells, the proliferation and migration ability of activated macrophages and the like. The seven feature dimensions of the physiological structure feature of the skin model are used for screening the to-be-tested sample and exploring the action mechanism, and the method has the characteristics of rapidness, multiple screening dimensions, high accuracy, low construction difficulty, low cost and high universality.

The invention discloses a co-culture method of gastric cancer organoid and gastric cancer-related myofibroblast and application of the co-culture method, gastric cancer organoid single-cell suspension and gastric cancer-related myofibroblast single-cell suspension are mixed according to the number ratio of 1: 1, then matrigel is added, after uniform mixing, the mixture is put into a CO2 cell incubator to solidify the matrigel, and the gastric cancer-related myofibroblast co-culture medium is obtained. The co-cultured gastric cancer organoid is faster in growth speed and stronger in resistance to chemotherapeutic drugs, the drug resistance of tumor cells in the body of a gastric cancer patient to the chemotherapeutic drugs is better simulated, and the co-cultured gastric cancer organoid can become a screening platform of individualized therapeutic drugs for tumor patients.

The invention relates to the technical field of plant genetic engineering, in particular to a genetic transformation method of prunus mume line plum blossom leaf calluses. The method comprises the following steps: firstly preparing a plum blossom genetic transformation receptor, then transforming agrobacterium tumefaciens by taking an RUBY gene as a reporter gene to prepare an infection bacterium solution, then carrying out callus infection and co-culture, and finally screening to obtain the resistant callus. According to the method provided by the invention, the resistant callus can be identified in an observation or PCR (Polymerase Chain Reaction) detection mode. The plum blossom callus is successfully infected, the red callus expressed by the RUBY reporter gene is obtained, the inductivity is high, and a good basis is provided for establishment and genetic improvement of a plum blossom genetic system.

The invention belongs to the technical field of organoid culture, and particularly relates to a co-differentiated vascularized islet organoid and a construction method thereof. Aiming at the problems that the existing pancreatic islet organ vascularization mostly adopts a co-culture mode, the operation process is long, the difficulty is high, the randomness is realized, the uniformity is poor and the like, the invention provides the construction method of the co-differentiated vascularized pancreatic islet organ. The method comprises the following steps of: sequentially inducing and culturing directional endoderm cells, original intestinal canal cells and posterior preintestinal cells, and adding a culture medium containing VEGF (vascular endothelial growth factor), BMP4 (bone morphogenetic protein 4) and FGF2 (fibroblast growth factor 2) for culturing in subsequent induced differentiation, so as to obtain the co-differentiated vascularized islet organ. Under the condition that exogenous endothelial cells are not added, the iPS cells are directionally differentiated to generate the vascularized islet organ containing beta, alpha and delta cells and vascular endothelial cells for the first time, the vascularized islet organ is more mature than non-vascularized islet organs and has a stronger insulin secretion function, and a new thought and an experimental basis are provided for realizing functional healing of diabetes mellitus.

The invention belongs to the technical field of biotechnology and animal models, and discloses a double-person-derived mouse model for simulating a tumor immune microenvironment and a construction method and application thereof. The method comprises the following steps: firstly, pretreating NSG immunodeficient mice by adopting low-dose whole-body irradiation in combination with double-antibody targeted bone marrow depletion, and transplanting CD34 + hematopoietic stem cells from the same human donor to complete human immune system reconstruction; separating tumor primary cells, tumor-related fibroblasts and tumor vascular endothelial cells of the same donor, performing three-dimensional co-culture to obtain homologous human tumor organs, and performing in-situ inoculation to immune reconstruction mice to obtain a target model. The core defects of MHC mismatching, low immune reconstruction efficiency, poor tumor immune microenvironment simulation degree, low clinical consistency and the like of an existing model are overcome, and the method can be used for tumor immune treatment drug screening, microenvironment mechanism research and personalized tumor treatment scheme verification.

The invention provides an organ chip unit for simulating the pathogenesis of rheumatoid arthritis, and belongs to the technical field of cell culture. The chip comprises a top-layer chip, a bottom-layer chip and a middle microporous membrane, the top-layer chip and the bottom-layer chip are separated by the middle microporous membrane, and the middle microporous membrane is used for enabling cells in a cell culture space of the top-layer chip and cells in a cell culture space of the bottom-layer chip not to be in direct contact, but interaction of signal molecules and substance exchange can be carried out; synovial cells and cartilage cells are injected into the bottom-layer chip or the top-layer chip to be cultured. According to the invention, multi-cell three-dimensional co-culture can be realized. Through the additional stress stimulation co-culture part, the influence of pressure or tension on the cell environment can be judged, and the actual condition that rheumatoid arthritis joints receive pressure or tension for a long time can be simulated. The synovial cells and the cartilage cells are used, so that the structure of the inflammation joint in a living body can be truly reflected, and the interaction among various cell tissues can be reflected.

The invention relates to the technical field of biology, in particular to a Korean pine control-based method for researching the resistance mechanism of larch to pine wood nematodes, which comprises the following steps: S1, extracting pine wood nematodes: separating to-be-tested nematodes from naturally infected larch by using a funnel; s2, pine wood nematode culture: utilizing botrytis cinerea to co-culture pine wood nematode as a test raw material; s3, selecting tree species: selecting twigs, boughs and trunks of healthy larch with tree ages of 1 year, 2 years and 3 years respectively, and selecting Korean pine with the same requirements as a contrast; s4, sampling tree species: sampling larch and Korean pine inoculated for 1 day, 3 days, 5 days, 7 days, 9 days, 14 days and 21 days; by analyzing the chemical components of the Korean pine and the larch in different stages before, during and after the bursaphelenchus xylophilus infection, the purpose of researching the pathogenesis of the Korean pine and the larch by local pine nematodes in Liaoning is achieved, so that the purpose of effectively preventing the bursaphelenchus xylophilus is achieved.