49 results about "Karyotype" patented technology

The invention provides a full-automatic indirect immunofluorescence interpretation method and system, a storage medium and a program product, and relates to the technical field of indirect immunofluorescence detection.The method comprises the steps that when identification information of a target sample is obtained, a plurality of sealed antigen slides are placed in slide carrying groove positions of a bearing platform in batches, and the target sample is obtained; driving the bearing platform through the transmission mechanism to sequentially move the plurality of antigen slides to the imaging area, and performing positioning and fixing operation on the antigen slides entering the imaging area by using the positioning and clamping device; sequentially carrying out multi-view scanning on the antigen slides entering the imaging area to obtain multi-view fluorescence images, and splicing the fluorescence images to generate a fluorescence panoramic image; performing feature extraction on the panoramic image, determining a fluorescence karyotype category and a target fluorescence intensity value, and determining an antibody titer according to the category, the intensity value and dilution information; and performing association mapping on the titer, the category and the panoramic image according to the identification information to generate a fluorescence interpretation result.

The invention provides a chromosome image enhancement method and system based on semantic guidance. The method comprises the following steps: S1, preprocessing; s2, outputting a deep-band probability graph, a grey-band probability graph and a shallow-band probability graph through a semantic segmentation network composed of a lightweight encoder and a multi-scale decoder; s3, implementing differential layered enhancement according to a band type: adopting local adaptive histogram equalization for a deep band, adopting central axis constraint bilateral filtering for a gray band, adopting dynamic threshold truncation and gamma correction for a shallow band, and performing weighted fusion for a transition region according to probability; s4, carrying out structure strengthening, wherein the structure strengthening comprises centromere local sharpening, stripe phase alignment, edge sensing super-resolution and overlapping region separation; and S5, performing quality evaluation based on the deep band integrity, the band stripe contrast uniformity, the SSIM and the noise density, and triggering adaptive re-enhancement if necessary. According to the scheme, the contrast ratio and details are remarkably improved while the stripe structure and the position relation are kept, and the method has the advantages of light weight, interpretability and cross-sample robustness and is suitable for being integrated into an automatic karyotype analysis process.

The invention discloses a chromosome image generation method and device for deep learning. Performing adaptive multi-scale image enhancement processing on the chromosome mid-term image to obtain an enhanced chromosome mid-term image; obtaining a chromosome karyotype arrangement diagram based on the enhanced chromosome metaphase image; generating a chromosome set based on the chromosome karyotype arrangement diagram; sequentially merging the chromosomes in the chromosome set into one image as a chromosome foreground image; fusing the chromosome foreground image with a randomly selected background image, and carrying out edge smoothing processing on the fused image to generate a random background chromosome middle-term image; wherein the random background chromosome metaphase image is used for training a target detection model established based on deep learning. According to the method, high-quality and diversified chromosome metaphase images can be generated, and when the method is used for training the deep learning model, the generalization ability and precision of the deep learning model in a target detection task are improved.

The application discloses a construction method and application of an ovary cell line of Quasipaa spinosa. The method comprises the following steps: selecting metamorphosis period larva ovary tissue, disinfecting and obtaining the ovary tissue, and then using collagenase I digestion solution to digest the ovary tissue at 26 DEG C for 1 hour; after the tissue block adheres to the wall, using a low-osmotic-pressure complete culture medium to carry out primary culture at 25-27 DEG C, wherein the culture medium is 60% diluted DMEM / F12 culture medium containing 10%-15% fetal bovine serum; when the cell confluence degree is higher than 70%, using trypsin-EDTA to digest the cells for 1-2 minutes to subculture, and the subculture culture medium contains 30% cell conditional culture medium; and finally, the cells are cryopreserved through programmed cooling. The ovary epithelial cell line of Quasipaa spinosa is successfully established for the first time, the survival rate of the cells after recovery is high, and the karyotype is stable. The constructed cell line can be used for amphibian reproductive biology research, compound screening and in-vitro conservation of germplasm resources.

The invention discloses a chromosome karyotype analysis method and system, and relates to the technical field of image recognition, and the method comprises the following steps: S1, obtaining to-be-analyzed chromosome image data, and marking the to-be-analyzed chromosome image data to obtain a to-be-analyzed image data set; s2, introducing an improved module to perform network reconstruction on the YOLOv11 network model to obtain a chromosome detection model; s3, training a chromosome detection model through the to-be-analyzed image data set, and performing performance evaluation to obtain a target chromosome detection model; and S4, analyzing the real-time chromosome image data through the target chromosome detection model, and outputting an analysis result. According to the scheme, the detection capability of the model on chromosomes in small-target, abnormal and complex scenes is enhanced by utilizing an improved module, and the problems of low accuracy and poor robustness in the prior art are solved, so that efficient and accurate chromosome karyotype analysis is realized, and clinical and occupational health examination requirements are met.

A chromosome image segmentation identification method and system based on deep learning relates to the technical field of medical image processing, and comprises the following steps: performing target detection processing on original chromosome image data, and outputting chromosome image data calibrated with chromosome clusters; performing multi-level convolution and down-sampling processing on the chromosome image data by using a dynamic convolution kernel to obtain middle-layer down-sampling feature data and final-layer down-sampling feature data; in combination with a parameter-free attention mechanism, performing depth separable convolution processing and pixel recombination processing on the sampling feature data under the final layer to obtain deep feature optimization data; performing deconvolution processing and jump connection splicing processing on the deep feature optimization data and the intermediate layer down-sampling feature data, and outputting chromosome segmentation image data; classifying and arranging the chromosome segmentation image data to obtain a chromosome karyotype analysis result; the chromosome image segmentation and identification method and system have good segmentation and identification effects on the chromosome image.

The application belongs to the technical field of biology, and discloses a use of antisense nucleotide of piR-bmo-796514 nucleotide for preparing a medicine for inhibiting Bombyx mori nuclear polyhedrosis virus, the antisense nucleotide of the piR-bmo-796514 nucleotide is mainly used for inhibiting replication of the Bombyx mori nuclear polyhedrosis virus in ovary cells of the Bombyx mori, and a sequence of the piRNA nucleotide is shown as SEQ ID NO:1; in addition, the application also discloses the antisense nucleotide of the piR-bmo-796514 nucleotide, and an antisense nucleotide sequence is shown as SEQ ID NO:2.

The invention discloses an SSR molecular marker for screening nuclear cracking resistance characters of peaches and application of the SSR molecular marker, and relates to the technical field of genetic breeding of fruit trees, the SSR molecular marker is named as PpCPK12-SSR, and the nucleotide sequence of the SSR molecular marker is shown as SEQ ID NO.1. The invention also provides a specific primer pair for amplifying the marker, the nucleotide sequence of an upstream primer of the specific primer pair is shown as SEQ ID NO.2, and the nucleotide sequence of a downstream primer of the specific primer pair is shown as SEQ ID NO.2. The nucleotide sequence of the downstream primer is as shown in SEQ ID NO.3. Through PCR (polymerase chain reaction) amplification and electrophoresis detection, according to the size of an amplified fragment, the anti-nuclear-cleavage genotype of the peach can be quickly and accurately identified in the seedling stage of the peach: an individual carrying a 115bp fragment is an anti-nuclear-cleavage type; the defects that traditional field phenotype identification is long in period, low in efficiency and affected by the environment are overcome, an efficient molecular marker auxiliary selection tool is provided for peach nuclear cracking resistance breeding, and the breeding process can be remarkably accelerated.

The invention provides a novel human distal bile duct cancer cell line with TP53 missense mutation and application, the novel human distal bile duct cancer cell line CBC3T-3 is established, the cell line is preserved in the China Center for Type Culture Collection (the preservation number is CCTCC NO: C202555), the uniqueness and stability of the cell line are proved through STR typing and karyotype analysis, and the TP53 missense mutation novel human distal bile duct cancer cell line has the advantages that the TP53 missense mutation novel human distal bile duct cancer cell line CBC3T-3 can be used for preparing the TP53 missense mutation novel human distal bile duct cancer cell line CBC3T-3; and a plurality of driver gene mutations including TP53 missense mutation are carried. The CBC3T-3 has strong proliferation, invasion and migration capabilities, has high tumor formation rate in immunodeficient mice, is resistant to cis-platinum and sensitive to paclitaxel and gemcitabine, and provides an experimental basis for selection of clinical chemotherapy regimens. According to the model, the TP53 missense mutation type distal bile duct cancer in-vitro model is successfully established, and a key experimental platform is provided for deeply researching the drug resistance mechanism of the TP53 missense mutation type distal bile duct cancer and developing an individualized treatment strategy aiming at the subtype of the TP53 missense mutation type distal bile duct cancer.

The invention discloses a quality control product for noninvasive prenatal detection of fetal chromosome trisomy syndrome and a preparation method thereof, the quality control product comprises a positive quality control product and a negative quality control product, the positive quality control product is obtained by adding nucleic acid fragments of trisomy syndrome genome DNA into background plasma; the negative quality control product is obtained by adding nucleic acid fragments of normal karyotype genome DNA into background plasma; the length of the nucleic acid fragment is 130 to 200 bp; the background plasma is peripheral blood plasma of normal karyotype and pregnant fetus women. According to the invention, peripheral blood plasma of a pregnant woman with normal karyotype and pregnant fetus is used as background plasma, a DNA extraction correction coefficient is introduced, and nucleic acid fragments of trisomatic negative and positive genomes 130-200bp with sex chromosome XY are added to prepare the quality control product, so that the quality control product is more accurate and reliable, and is more suitable for pregnant woman plasma; and whether each index of the noninvasive antenatal detection kit meets the technical requirements of a product or not can be reflected more completely and truly.

The invention provides a gene with an antiviral effect and application thereof, the nucleotide sequence of an open reading frame of the gene is SEQ ID NO: 2, and the amino acid sequence of a coding protein of the gene is SEQ ID NO: 1. The invention also provides a method for increasing the sensitivity of the beet armyworm to the beet armyworm nuclear polyhedrosis virus, which is used for reducing the expression quantity of the gene in the beet armyworm. According to the invention, a gene with an antiviral effect is screened from an important agricultural pest beet armyworm. The sequence structure and tissue expression mode of the gene are analyzed, incremental expression and RNA interference technologies are utilized to prove that the gene has an anti-SeMNPV effect, a new target is provided for better utilizing baculovirus to prevent and control beet armyworms in production, and the gene has a good application prospect.

The invention provides an improved euchiloglanis kishinouyei chromosome preparation method, which comprises the following steps: gradually raising the temperature and injecting PHA twice: in the euchiloglanis kishinouyei temporary culture process, raising the water temperature from 18 DEG C to 20 DEG C within 12 hours, injecting a first needle of PHA, raising the temperature to 22.5-23 DEG C after 12 hours, injecting a second needle of PHA, and keeping the water temperature at 22.5-23 DEG C unchanged; after the PHA is injected for the second time for 3 hours, colchicine is injected; sampling: washing the head and kidney in normal saline, and collecting cells; 0.5% of KCl is used for hypotonic treatment; fixing with a Carnoy's fixing solution, and centrifuging; dropping by a cold dropping method and naturally drying; and carrying out Giemsa dyeing and flaking. According to the method, the euchiloglanis kishinouyei somatic cell proliferation is promoted through heating and twice PHA injection, so that the metaphase division phase is increased, the form is clear and dispersed, and the karyotype quality and the preparation success rate of the chromosome are remarkably improved.

Methods and techniques for controlled printing of a cell sample for karyotyping are provided. The methods can involve matrix printing using on-the-fly printing or dispensing to accurately spread cells within at least one cell sample on a surface in preparation for karyotyping, and further analysis. Advantageously, the methods result in a uniform distribution of chromosomes of the cell suspension or sample on the surface of a substrate which can be substantially discretely identified, and also provide for efficiency in a subsequent staining process and any further analysis of the stained chromosomes using a microscope or other imaging device.

The present application relates to the field of aquatic organism breeding technology, and particularly relates to a method for preparing rainbow trout tetraploid fry. The method provided by the present application comprises the following steps: obtaining fertilized eggs by dry fertilization of naturally matured rainbow trout parents, inducing the rainbow trout fertilized eggs to obtain tetraploid rainbow trout after 6.2-6.4 hours, and the duration of the induction is 10-18 minutes; after the tetraploid rainbow trout fry is obtained by induction, the chromosome ploidy of the fish is determined by using techniques such as chromosome karyotype analysis and flow cytometry DNA content determination, and the tetraploid rainbow trout fry is identified and screened for cultivation. The present application adopts a drug induction method, and controls the starting time and duration of the induction to inhibit the first mitosis time of the fertilized eggs, thereby improving the induction rate and survival rate of the tetraploid rainbow trout, and the induction rate of the tetraploid rainbow trout can reach 25-30%, and the survival rate of the fry can reach more than 90%.

The invention belongs to the technical field of biological medicine, and discloses a safe and efficient non-gene editing neural stem cell and application thereof in treatment of nervous system related diseases. The invention provides a non-gene-edited neural stem cell (hCiPSC-NSCs) derived from human induced pluripotent stem cells, the neural stem cell is obtained by reprogramming human umbilical cord mesenchymal stem cells through a small molecule compound (such as a TGF-beta / GSK-3beta inhibitor), exogenous gene introduction is not involved, and the neural stem cell has a normal karyotype, no tumorigenicity and relatively high safety; and the efficient implantation of the non-tumorigenicity hippocampal region is realized in three months. The neural stem cell can improve the autism-like phenotype of a VPA-exposed rat by targeting the intestinal-brain axis at the same time.

The application discloses a human lung large cell neuroendocrine carcinoma cell line and application, the cell line is preserved in China typical culture preservation center, the preservation time is August 22, 2024, the preservation number is CCTCC NO:C2024278, and the culture name is human lung large cell neuroendocrine carcinoma cell strain ZX2024C.The cell line can be stably passed, can keep the biological characteristics of human lung large cell neuroendocrine carcinoma cell, no human cell cross contamination is found, and has abnormal chromosome karyotype, can form tumor in the body of nude mouse, can be applied to the preparation of lung large cell neuroendocrine carcinoma diagnostic product, screening lung large cell neuroendocrine carcinoma drug, researching lung large cell neuroendocrine carcinoma cell proliferation mechanism, constructing mammalian lung large cell neuroendocrine carcinoma animal model.

The invention relates to the technical field of cell separation and purification and cell biological detection, in particular to a method for separating and purifying fetal NRBC from peripheral blood of a pregnant woman. The invention provides a method for separating and purifying fetal NRBC from peripheral blood of a pregnant woman, which is characterized in that a LukSF and LukAB bi-component perforation toxin is designed and utilized to specifically recognize and split a CXCR1 / 2 chemotactic factor receptor and a CD11b / CD18 integrin receptor on the surface of maternal leukocyte, and the fetal NRBC is completely reserved due to lack of corresponding receptors, so that the fetal NRBC can be separated and purified. Therefore, leukocyte pollution in nucleated red blood cells of a peripheral blood fetus of a pregnant woman is efficiently removed. According to the method, the purity of the extracted fetal NRBC can be improved, the false positive of gene detection is greatly reduced, the activity and integrity of the NRBC are reserved, and the accurate diagnosis requirements of chromosome karyotype analysis, monogenic disease detection and the like are met; the method is simple to operate and low in cost, can be directly adapted to existing clinical laboratory equipment, and has large-scale popularization value.

The invention discloses a construction method and application of an ovarian cell line of hapalogenys nitens. The method comprises the following steps: selecting a metamorphosis stage larva ovarian tissue, disinfecting and obtaining, and digesting for 1 hour at 26 DEG C by using collagenase I digestive juice; after the tissue block adheres to the wall, a low-osmotic-pressure complete medium is used for primary culture at the temperature of 25-27 DEG C, and the medium is a 60% diluted DMEM / F12 medium containing 10%-15% of fetal calf serum; after the cell confluence degree exceeds 70%, trypsin-EDTA (Ethylene Diamine Tetraacetic Acid) is used for digesting for 1-2 minutes for passage, and a passage culture medium contains 30% of a cell condition culture medium; and finally performing programmed cooling and cryopreservation. According to the method, the ovarian epithelial cell line of the hapalogenys nitens is successfully established for the first time, the cell recovery survival rate is high, and the karyotype is stable. The constructed cell line can be used for amphibian reproductive biology research, compound screening and germplasm resource in-vitro preservation.

The invention discloses a chromosome counting method and device based on image processing and electronic equipment. The method comprises the steps that computer equipment acquires a chromosome karyotype graph; projecting each row of pixel points of the chromosome karyotype graph to obtain a first projection vector; determining a plurality of foreground regions of the chromosome karyotype graph according to the first projection vector; selecting a plurality of first foreground areas from the plurality of foreground areas, and extracting target image areas corresponding to the plurality of first foreground areas; projecting each column of pixel points of the target image area to obtain a second projection vector; determining one or more chromosome regions contained in the target image region according to the second projection vector; selecting a target chromosome region comprising the chromosomes of the target type from one or more chromosome regions contained in the target image region; and obtaining the chromosome number corresponding to the chromosome of the target type. According to the invention, the counting accuracy and efficiency of the chromosome karyotype graph can be improved.

The application relates to a chromosome image segmentation and recognition method and system based on deep learning, and relates to the technical field of medical image processing.The method comprises the following steps: performing target detection processing on original chromosome image data to output chromosome image data with labeled chromosome clusters; using a dynamic convolution kernel and performing multi-level convolution and down-sampling processing on the chromosome image data to obtain intermediate layer down-sampling feature data and final layer down-sampling feature data; combining a parameter-free attention mechanism, performing deep separable convolution processing and pixel recombination processing on the final layer down-sampling feature data to obtain deep layer feature optimization data; performing inverse convolution processing and jump connection splicing processing on the deep layer feature optimization data and the intermediate layer down-sampling feature data to output chromosome segmentation image data; and performing classification and arrangement processing on the chromosome segmentation image data to obtain a chromosome karyotype analysis result.The chromosome image segmentation and recognition method and system can achieve good segmentation and recognition effects on chromosome images.

The invention discloses a preparation method of chromosomes of tiny insects trichogramma dendrolimus, and belongs to the technical field of chromosome preparation. Comprising the following steps: (1) preparing a pre-pupa material: selecting corcyra cephalonica eggs parasitized by trichogramma dendrolimus for 4-8 days, and cutting out a trichogramma dendrolimus pre-pupa; (2) pretreatment: pretreating the pre-pupa material in a colchicine solution; (3) low-permeability treatment: carrying out low-permeability treatment on the pretreated material in a low-permeability solution; and (4) fixing, uncovering, dyeing and sealing the material subjected to the low-permeability treatment. The invention provides a simpler trichogramma dendrolimus chromosome slide preparation method, chromosome slides with clear forms can be prepared in batches, the dispersion degree of chromosomes is large, the length, centromere position, arm ratio and other characteristics of the chromosomes can be observed more clearly, the chromosome characteristics are determined through karyotype analysis, and the chromosome slide preparation method is suitable for large-scale production. And a foundation is laid for cell level research and genomics deep research of trichogramma dendrolimus.

The application provides a miscarriage risk assessment method and system of whole-exome load and serum markers, comprising: independently scoring each variation in whole-exome sequencing data to obtain a comprehensive pathogenic score; constructing a gene pathogenic load matrix; mapping the gene pathogenic load to a biological pathway to obtain a pathway comprehensive score and construct a model; solving the model by jointly optimizing the regularization strength parameters of the two-layer ridge penalty terms, outputting significant pathway features; standardizing and preprocessing serum detection data; jointly modeling the output significant pathway features and the output standardized serum data under the same logistic regression probability framework, outputting a miscarriage-related risk probability value, and performing probability stratification. The application also outputs a double-track coordination coefficient to quantify the directional consistency of genetic signals and serum signals. It is significantly superior to a single omics solution and provides a high-precision individualized risk assessment tool for recurrent miscarriage with normal karyotype.

The invention discloses application of an METTL14 inhibitor in preparation of a medicine for treating malignant tumors carrying DNA outside chromosomes, and belongs to the technical field of medicines. A human colon cancer cell HT29-4 containing ecDNA and an ovarian cancer cell UACC-1598-4 are selected as research objects, an METTL14 stable knock-down and overexpression cell model is constructed through a lentivirus-mediated RNA interference technology, and the knock-down and overexpression efficiency is verified through Western Blot. Kyotype analysis shows that the quantity of ecDNA is reduced by knocking down the METTL14, and the quantity of the ecDNA is increased by overexpressing the METTL14. The copy number of DHFR and EIF5A2 genes carried by ecDNA is detected through qPCR, and the result shows that the copy number is decreased after METTL14 is knocked down, and the copy number of overexpression groups is increased. EdU proliferation experiments and cell scratch experiments show that knock-down of METTL14 inhibits proliferation and migration ability of tumor cells containing ecDNA, and overexpression promotes the phenotypes. The invention provides an effective treatment means for the intractable tumor driven by the ecDNA.

The invention relates to a conditional gene knockout / knockin animal model construction method based on a Cre-ES cell system. Firstly, an embryonic stem cell line (Cre-ES) capable of stably expressing Cre recombinase is established from a blastocyst of a tissue-specific Cre tool mouse, the normal karyotype rate of the embryonic stem cell line is larger than or equal to 85%, Cre genes are stably integrated, the chimeric rate after blastocyst injection reaches 100%, and the birth rate is larger than 20%. By utilizing the cell line, a gene editing carrier (such as a loxP-Stop-loxP report system targeting ROSA26) containing a loxP site is introduced into a cell through electrotransfection, positive clones are obtained through G418 screening and then injected to an ICR receptor blastocyst, and an F0-generation gene editing animal is directly obtained. The genotype of the F0-generation animal is flox / flox, and the F0-generation animal carries Cre recombinase, so that tissue specific editing can be realized without mating with a tool mouse. According to the method, the traditional four-generation breeding period is shortened to one generation, the efficiency is improved by 70%, the cost is reduced by 50%, the problems of line mixing, Cre leakage expression and marker gene residue are solved, and the method is suitable for space-time specific gene function research and disease model construction.

The application discloses a chromosome image generation method and device for deep learning. Adaptive multi-scale image enhancement processing is performed on a chromosome metaphase image to obtain an enhanced chromosome metaphase image; a chromosome karyotype arrangement map is obtained based on the enhanced chromosome metaphase image; a chromosome set is generated based on the chromosome karyotype arrangement map; chromosomes in the chromosome set are sequentially merged into one image as a chromosome foreground image; the chromosome foreground image is fused with a randomly selected background image, and edge smoothing processing is performed on the fused image to generate a random background chromosome metaphase image; wherein the random background chromosome metaphase image is used for training a target detection model established based on deep learning. The method can generate high-quality and diversified chromosome metaphase images, and thus is used for training a deep learning model, and improves the generalization ability and precision of the deep learning model in a target detection task.

The invention discloses establishment and application of a humpback perch frontal bone tissue osteoblast line, and relates to the technical field of biology. The cell line is a humpback perch frontal bone tissue osteoblast line CAFBOB, the preservation number is CCTCC NO: C2025129, and the cell line is derived from a humpback perch juvenile fish frontal bone tissue and is established after tissue separation, primary culture by a dry pasting method, subculture and cryopreservation resuscitation. The biological characteristics are stable, 1-40 generations are mainly long fusiform, the chromosome karyotype is 2n = 48, strong alkaline phosphatase activity is achieved, the transfection efficiency is about 85%, and the activity is good after cryopreservation and resuscitation. The establishment method is high in repeatability, the obtained cell line can be used as an in-vitro research model and applied to morphological specialization mechanism analysis, osteogenesis-related gene function verification, classification dispute solution and seeding process research of the humpback perch, the blank of the bone tissue cell line of the humpback perch is filled, and the bone tissue cell line has important scientific research value and application prospect.

The invention discloses a macaque skin squamous carcinoma cell line MCSCC14397 and application of the macaque skin squamous carcinoma cell line MCSCC14397. The macaque skin squamous carcinoma cell line is named as macaque skin squamous carcinoma cell line MCSCC14397, and the preservation number of the macaque skin squamous carcinoma cell line is CCTCC (China Center For Type Culture Collection) NO: C2025156. The cell line disclosed by the invention has similar protein expression with primary tumor tissues, has chromosome abnormal karyotype, in-vitro tumor formation capability and stable proliferation characteristics, and simultaneously shows remarkable genome instability (such as high-frequency Cgt, T mutation and structural variation) and inflammation signal activation. The MYO10 gene expression in the cell line is up-regulated and drives DNA damage reaction and inflammatory factor expression, and unique advantages are provided for application of the cell line in skin squamous cell carcinoma mechanism research, drug screening and drug efficacy evaluation. The cell line disclosed by the invention can be used as an effective cell model, can be used for establishing a xenotransplantation animal model, and can provide a research basis for deeply researching the occurrence, development, metastasis mechanism and drug resistance mechanism of human skin squamous cell carcinoma as well as innovative drug development and screening.

The invention discloses a bovine ectoderm stem cell line as well as an establishment method and application thereof. The bovine ectoderm stem cell line has the pluripotency of a bovine intermediate state stem cell line, expresses one or more pluripotency markers, can be stably passaged, still maintains pluripotency transcriptome characteristics and normal karyotype similar to those of an intermediate state (Formative) ectoderm cell after more than 112 generations, and has the potential of being differentiated into three-germ layers. The bovine intermediate-state stem cell line has the potential for myogenic differentiation, primordial germ cell-like cell differentiation and serving as somatic cell nuclear transplantation (SCNT) donor cells, which shows that the bovine intermediate-state stem cell line has wide application prospects in the aspects of promoting cell culture meat production and animal breeding.

An improved baculovirus expression system for enhanced heterologous protein expression in hosts selected from Bombyx mori larvae, pupae, primary cells, or established insect cell lines is disclosed. The expression system is derived from a wild-type strain of Bombyx mori nucleopolyhedrovirus (BmNPV) by deletion or inactivation of non-essential genes, of the BmNPV genome. The system includes novel transfer vectors designed for targeted gene modification and incorporation of a bacterial origin of replication and an insertion site for a gene of interest (GOI). This engineered bacmid maintains infectivity while offering enhanced genetic stability and significantly improved heterologous protein expression in insect cells, larvae, or pupae. Methods for constructing this bacmid via homologous recombination and using it for high-level protein production are also described.

The invention discloses establishment and application of an epinephelus lanceolatus frontal bone tissue osteoblast line, and relates to the technical field of biology. The cell line is an epinephelus lanceolatus frontal bone tissue osteoblast line ELFBOB, the preservation number is CCTCC NO: C2025132, the cell line is derived from epinephelus lanceolatus fry frontal bone tissue, and the cell line is established after tissue separation, primary culture with a dry pasting method, subculture and cryopreservation resuscitation. The biological characteristics are stable, 1-40 generations are mainly long fusiform, the chromosome karyotype is 2n = 48, strong alkaline phosphatase activity is achieved, the transfection efficiency is about 75%, and the activity is good after cryopreservation and resuscitation. The establishment method is high in repeatability, the obtained cell line can serve as an in-vitro research model and is applied to epinephelus lanceolatus skull malformation cause analysis, osteogenesis-related gene function verification, improved variety breeding and healthy culture strategy optimization, the blank of the epinephelus lanceolatus bone tissue cell line is filled, and the method has important scientific research value and industrial application prospects.