50 results about "Neonatal rat" patented technology

The invention provides the construction for a farnesyl pyrophosphoric acid synthetase RNA (Ribonucleic Acid) interference recombinant lentivirus vector, which comprises the following steps of: sieving the most effective target sequence of an FDS (farnesyl diphosphate synthase) gene RNAi (RNA interference) in a tool cell 293T cell, synthesizing the double-stranded DNA of the most effective target sequence, connecting to a pGCSIL-GFP vector and successfully constructing the recombinant vector through enzyme cutting, sequencing and identification. Researches indicate that the constructed RNA interference vector LV-sh-FDS can downwards modulate the expression of an FDS mRNA (Messenger RNA) level in a neonatal rat cardiac myocyte, simultaneously can downwards modulate the expression of myocardial hypertrophy markers such as cell areas and marker genes beta-MHC (Myosin Heavy Chain) and BNP (Brain Natriuretic Peptide), additionally can effectively inhabit the activity of RhoA while downwards modulating the FDS, can be applied in preparing medicaments for treating myocardial hypertrophy diseases and also can be applied in preparing medicaments for cholesterol metabolic control.

The invention discloses an application of a guanidino compound represented by formula I or a pharmaceutically acceptable salt of the guanidino compound in the preparation of medicines for treating nervous system degenerative diseases. R in the formula I is hydrogen, a carboxyl group or a -C1~C5 alkyl-carboxyl group. The guanidino compound has a substantial protection effect on hydrogen peroxide induced neonatal rat hippocampal nerve cell injures, has a substantial inhibition effect on in vitro beta-amyloid protein aggregation, and has potential clinic application values.

The invention discloses a method for separating and purifying oligodendrocyte precursor cells. The method comprises: digesting cerebral cortex of neonatal rats and part of white matter with trypsin and DNA enzyme; blowing and beating the obtained product to be a single-cell suspension; inoculating the single-cell suspension into a culture flask coated with polylysine in advance by use of a mixed-cell culture medium; performing culture for 3 to 5 days; replacing the culture flask with an oligodendrocyte precursor cell proliferation culture medium when the fusion of bottom-layer cells reaches 65 to 75 percent; performing culture for 3 to 5 days; replacing the oligodendrocyte precursor cell proliferation culture medium with an oligodendrocyte precursor cell separation culture medium; digesting the obtained product at 37 DEG C; isolating oligodendrocyte precursor cells from mixed cells; blowing and beating the obtained product to be the single-cell suspension; inoculating the single-cell suspension into the culture flask coated with polylysine in advance by use of an oligodendrocyte precursor cell inoculation culture medium; replacing the culture flask with an oligodendrocyte precursor cell purification culture medium after the cells adhere to a wall; performing culture for 2 to 4 days; and obtaining adherence cells, namely the oligodendrocyte precursor cells. The method can obtain a large number of high-purity good-activity oligodendrocyte precursor cells conveniently, rapidly, economically and efficiently.

Described herein are compositions and methods related to use of cardiosphere-derived cells and their extracellular vesicles, such as exosomes and microvesicles, for achieving anti-aging and rejuvenation. This includes discoveries for effects on heart structure, function, gene expression, and systemic parameters. For animal studies, intra-cardiac injections of neonatal rat CDCs was compared to in old and young rats including evaluation of blood, echocardiographic, haemodynamic and treadmill stress tests. For in vitro studies, human heart progenitors from older donors, or cardiomyocytes from aged rats were exposed to human CDCs or cardiosphere derived cell (CDC) derived exosomes (CDC-XO) from pediatric donors. CDCs and CDC-XOs were capable of effectuating youthful patterns of gene expression in the hearts of old, along with a variant of physiological and function benefits, including elongation of telomere length. Together, these results indicate capacity of CDCs and CDC-XO to ward off the effects of aging through rejuvenation.

The invention discloses RNA and application of RNA in diseases of a cardiovascular system. According to RNA, the nucleotide sequence of RNA is a sequence 3 in a sequence table. The invention further provides a gene,encoding RNA. The nucleotide sequence of the gene of RNA is a sequence 4 in the sequence table. The experiment of the invention shows that RNA and the application of RNA in the diseases of the cardiovascular system find that by an induction of isoproterenol (ISO), RNAi method Knock down IHP-55 is used in neonatal rat cardiac fibroblasts to obviously promote proliferation of cardiac fibroblasts; ISO induced activation and kinase activity of P38MAPK can be enhanced in the Knock-down HIP-55 cardiac fibroblasts; and HIP-55 is a new protein which has a protective effect on cardiac fibrosis, and provides a new target for clinical diagnosis, treatment and new medicament development.

The present invention relates to the field of biotechnology and in particular to a construction method of an experimental animal model, specifically a mouse model, with non-alcoholic steatohepatitis (NASH) transforming into hepatic carcinoma (HCC), and use thereof. The present invention provides a construction method of a mouse model with NASH and chronic steatohepatitis developing into chronic hepatic fibrosis, cirrhosis and HCC. The construction method comprises the following steps: newborn C57BL / 6 mice (including male and female newborn mice) are subjected to subcutaneous single injection of streptozotocin (STZ) (100-400 [mu]g / newborn mouse) and the mice begin to be fed with experiment use high-fat feed after the completion of female mouse breastfeeding to construct the NASH mouse model with hepatic fibrosis, cirrhosis and HCC. The provided mouse model is relatively simple in construction process, relatively simple in technical means, and low in construction cost, has an extremely high value as the needed animal model in scientific research and drug research and development, and belongs to a domestic initiative.

The invention discloses a construction method of an in-vitro model for simulating mesenchymal stem cell transplantation, which comprises the following steps of: (1) selecting in-vitro neonatal rat cortical neurons; (2) selecting in-vitro rat mesenchymal stem cells; (3) pre-processing the in-vitro rat mesenchymal stem cells, and inoculating in the neonatal rat cortical neurons for co-culture; and (4) after the co-culture, treating the rat mesenchymal stem cells and the neonatal rat cortical neurons by a light microscope, a fluorescence microscope, an atomic force microscope and a scanning electron microscope; and determining an in-vitro model for simulating mesenchymal stem cell transplantation and a growth tendency, an interaction and formation of a synapsis-like structure between the cortical neurons and the mesenchymal stem cells on an ultrastructure. The construction method disclosed by the invention provides a simplified in-vitro model for the mesenchymal stem cell transplantation; meanwhile, the synapsis-like structure formed in the model provides an in-vitro evidence of a structure integration mechanism for the mesenchymal stem cell injury mechanism.

The invention discloses an application of a morindae officinalis extract and morindae officinalis oligosaccharide pentasaccharide to preparation of a drug for treating myocardial ischemia and / or reperfusion injury and promoting therapeutic angiogenesis. The myocardial ischemia and reperfusion injury belong to secondary injury brought to an organism when the hemoperfusion of myocardium is stopped or poor, i.e., after the myocardium is subjected to ischemia, hypoxia injury, circulation reinstitution and blood supply recovery. The morindae officinalis extract and the morindae officinalis oligosaccharide pentasaccharide both play a role in protecting an in-vivo rat myocardial ischemia reperfusion injury model and an in-vitro purification cultured neonatal rat myocardial cell hypoxia / reoxygenation injury model, and can be used for remarkably reducing the myocardial infarction area and the incidence rate of reperfusion arrhythmias, effectively protecting the form of a myocardial cell and relieving the injury of hypoxia / reoxygenation to the myocardial cell. The therapeutic angiogenesis promotion means that the aims of recovering the blood supply of ischemic myocardium and improving the heart function are achieved through increasing the functional coronary artery branch or side branch under the irritation actions of some methods, including the therapeutic actions of some drugs.

This invention relates to a safe carrier for the seed viruses of inactivated vaccine for the foot-and-mouth disease and the application thereof. Different serum-type seed viruses for producing the inactivated vaccines for the foot-and-mouth disease safely are constructed on the basis of the carrier. The constructed embedded viruses retain the advantage of good adaptability of the classic vaccine virus strain cell, overcome the defect of low matching performance of the antigen through the structural protein inserted into the epidemic virus strain and can save virus quickly to produce the inactivated vaccine for foot-and-mouth disease. The A or Asia I or O type different virus-strain embedded viruses which are constructed through the carrier are less virulent and have good bio-security. Because the viruses are short of the protein L, the virulence of the viruses can not become stronger under the natural condition, and the pathogenicity of the viruses on the neonatal rat, pig or cow is reduced greatly or the viruses are not pathogenic. The carrier is highly safe, can not cause the foot-and-mouth disease and is highly practical.

The invention discloses a primary culturing method for dorsal root ganglion satellite glial cells. The method comprises the operating steps of 1, killing a neonatal mouse or a neonatal rat after sterilization by breaking a head, taking out a spine after dissection, removing muscle on the spine, cutting off the spine, clearing blood vessels and spinal cord under a stereoscopic microscope and taking out DRG, and removing nerve fibers and envelopes on the DRG; 2, culturing the processed DRG by using DRG-SGCs culturing solution, inducing the large number of satellite glial cells to move out of the DRG, adding trypsin for digestion after culturing ends, then adding FBS for stopping digestion, slighting blowing and beating the solution by using a Pasteur pipet, and then transferring the solution into a centrifuge tube for centrifuging; and 3, discarding supernate after centrifuging, and then adding the culturing solution for continuous culturing, thus acquiring the satellite glial cells. The satellite glial cells cultured and purified according to the method provided by the invention can survive for a long time in vitro, is high impurity and stable in survival, and can be passaged for multiple generations and form a cell network.

The present invention discloses pharmaceutical use of Echinacoside (ECH) for treatment of hypoxic-ischemic encephalopathy. The experimental results of the present invention indicate that the Etoposideat a dose of 160 mg / kg can significantly improve cerebral infarction volume and neuronal damage and apoptosis in a hypoxic-ischemic model of neonatal rats when used in a safe dose range, and prove that the Etoposide can promote the recovery of neuronal ischemia and hypoxia injury and prevent disability and death after hypoxic ischemic encephalopathy.

The invention discloses a modeling method of irritable bowel syndrome. The method comprises the steps of using a balloon dilatation catheter to perform colonic expansion stimulation on newborn rats of8-21 days old, giving a pressure of 60 mmHg in the rectum for 1 min, and repeating the stimulation after 30 min. The abdominal wall retraction reflex test scoring is performed 90 days after birth, aresult that the AWR score of the model rats is significantly higher than that of a normal control group is obtained, and the result proves that the visceral hypersensitivity model is replicated successfully and continued to the adult stage of the rates, and histopathological examination, colon tissue SP immunohistochemical examination and other related factors demonstrate successful modeling of the method. The model of irritable bowel syndrome provided by the invention can be used for screening drugs for controlling irritable bowel syndrome, has good stability and is consistent with human IBSsymptoms, and thus can simulate a long-term chronic course of IBS.

Provided is a culture medium for the primary culture of hippocampus neurons of a neonatal rat. The culture medium is formed by adding 1,6-diphosphate fructose, fructose and nutritional supplements to a basic medium Neurobasal. Also provided is a method for the primary culture of hippocampus neurons of a rat using the abovementioned culture medium, comprising the steps of: (1) preparing the culture medium of hippocampus neurons; (2) sampling and digesting; (3) preparing a single cell suspension; (4) inoculating and culturing. Hippocampus neurons with good growing status and higher purity can be obtained using the abovementioned culture medium.

The invention provides a grouping purification method of olfactory ensheathing cells (OECs) of a neonatal rat, and the method is applied to a purification process of the OECs of the neonatal rat, wherein the purification process comprises the following steps of: obtaining the OECs; separating the OECs; performing grouping purification and comparison on the OECs; and performing morphologic observation and OECs purity detection. By virtue of different purification method experiments of the OECs of the neonatal rat, results indicate that the OECs of an in-vitro cultured neonatal rat are mainly bipolar or tripolar cells, and the neurite of the cell is fine. The OECs which are not purified have quickly-growing and dominated fibroblast, and the three purification methods realize that the purity average value of the OECs after being inoculated for 14 days is more than 75 percent. The purification rates of a p75 antibody adsorption method and a cytarabine inhibition method are slightly higher than that of a difference-speed adherence method. Therefore, cell purification in OECs primary culture of the neonatal rat is necessary; and in a spinal cord injury and regeneration research, compared with the p75 antibody adsorption method and the cytarabine inhibition method, the difference-speed adherence method is a simple, economical and practical OECs purification method.

The invention relates to a hemagglutination inhibition test antigen for a duck tembusu virus disease and a preparation method thereof. A duck hemorrhagic ovarian inflammation virus HB strain is preserved at the typical culture preservation centre in China with the preservation number being CCTCC V201122; the strain comprises a specific gene sequene, and the GenBank accession number of the strain is JF523187. The strain is inoculated againt a neonatal rat proliferating virus; the hemagglutination inhibition test antigen for the duck tembusu virus disease is prepared through the technologies of hemagglutination inhibition nonspecific material (lipoprotein) removal, inactivation, addition of a protective agent and the like. The invention further discloses the preparation method of the hemagglutination inhibition test antigen by a duck tembusu virus.

Oxygen toxicity is one of the major risk factors in the development of the chronic lung disease or bronchopulmonary dysplasia in premature infants. Using proteomic analysis, we discovered mitochondrial aldehyde dehydrogenase (mtALDH or ALDH2) was down-regulated in neonatal rat lung after hyperoxic exposure. To study the role of mtALDH in hyperoxic lung injury, we overexpressed mtALDH in human lung epithelial cells (A549) and found that mtALDH significantly reduced hyperoxia-induced cell death. Compared to control cells (Neo-A549), the necrotic cell death in mtALDH overexpressing cells (mtALDH-A549) decreased from 25.3% to 6.5%, 50.5% to 9.1% and 52.4% to 15.06% after 24-, 48- and 72-hour hyperoxic exposure, respectively. The levels of intracellular and mitochondria-derived reactive oxygen species (ROS) in mtALDH-A549 cells after hyperoxic exposure were significantly lowered compared to Neo-A549 cells. mtALDH overexpression significantly stimulated extracellular signal regulated kinase (ERK) phosphorylation under normoxic and hyperoxic conditions. Inhibition of ERK phosphorylation partially eliminated the protective effect of mtALDH in hyperoxia-induced cell death, suggesting ERK activation by mtALDH conferred cellular resistance to hyperoxia. mtALDH overexpression augmented Akt phosphorylation and maintained the total Akt level in mtALDH-A549 cells under normoxic and hyperoxic conditions. Inhibition of PI3K activation by LY294002 in mtALDH-A549 cells significantly increased necrotic cell death after hyperoxic exposure, indicating that PI3K / Akt activation by mtALDH played an important role in cell survival after hyperoxia. Taken together, these data demonstrate that mtALDH overexpression attenuates hyperoxia-induced cell death in lung epithelial cells through reduction of ROS, activation of ERK / MAPK and PI3K / Akt cell survival signaling pathways.

The invention discloses a method for determination of calcium concentration in a rat spinal dorsal horn neuron. The method includes: taking a SD neonatal rat, conducting disinfection and then executing it, separating the spinal dorsal horn tissue, stripping a spinal cord outer membrane, cutting the tissue into small pieces, transferring the pieces into a centrifuge tube, and adding trypsin to conduct digestion; then adding a DMEM / F12 medium containing 10% fetal calf serum to terminate the digestion; conducting blowing to obtain a cell suspension, performing centrifugation and discarding the supernatant; adding an NB / B27 medium to resuspend cells, inoculating the cells into a special laser confocal dish, and performing standing in an incubation box; adding an NB / B27 medium containing penicillin and streptomycin; performing loading with a fluorescent indicator, putting the cells into the incubation box to perform incubation, rinsing the NB medium, then covering the cells with the NB medium, and carrying out scanning detection with a laser scanning confocal microscope. The method for determination of calcium concentration in the rat spinal dorsal horn neuron provided by the invention can be used for observing the influence of corticosterone on the Ca<2+> concentration of the cultured spinal dorsal horn neuron, and creates conditions for studying the regulation of corticosterone on spinal dorsal horn sensory neuron functions and the action mechanism thereof.

The invention discloses an application of SAG in preparation of a drug for treating diseases of hypoxic-ischemic brain damage during development. Experiments show that intraperitoneal injection of 5-25mg / kg of SAG after 2 hours after hypoxic-ischemia in neonatal rats can reduce cerebral infarction. Further study on the neonatal rats applied with 15mg / kg of SAG shows that SAG can significantly improve brain damage and reduce nerve cell disintegration and necrosis after anoxia and ischemia. The Morris water maze experiments prove that 15mg / kg of SAG can effectively improve the learning ability of hypoxic-ischemic rats. Further study shows that 2, 3, 5-cholo-triphenyltetrazolium-positive cells of the rats in the SAG medium-dose experimental group rat (15mg / kg SAG) are significantly reduced, the IL-1 beta and TNF-alpha mRNA in the cerebral cortex are decreased and the expression of a glial fibrillary acidic protein is reduced.

The invention aims to provide a novel tropolone compound or a pharmaceutical salt of the tropolone compound, pharmaceutical composition taking the tropolone compound or the pharmaceutical salt as an active component, a preparation method of the tropolone compound, an application of the tropolone compound to preparation of drugs for treating or preventing myocardial cell damage as well as an application to preparation of antioxidants. The compound obtained with the method has very good antioxidant and anti-free-radical activity, has a good protection function for damage to H2O2 induced neonatal rat myocardial cells and can be used for preparing antioxidants and myocardial damage protective agents.

The invention discloses a method for increasing the reproduction rate of bamboo rats. The method includes the following steps that firstly, mating male rats and female rats are placed in a breeding pool to be bred according to the proportion of 1:1, wherein a breeding feed formula for every rat every day is prepared from 8-10 grains of corn, 45-55 g of fresh phyllostachyspubescens and 120-170 g of fresh pennisetumhydridum stems; secondly, the female rats are fed according to the feed formula in the first step after being fertilized, besides, 40-48 g of milk and 8-10 g of white sugar are fully dissolved in 45-55 g of warm boiled water and then put into a water trough so that the pregnant female rats can take the feed freely, and the female rats are fed once every day; thirdly, the female rats are fed according to the feed formula in the first step after littering and ablactation, besides, 40-48 g of milk and 8-10 g of white sugar are fully dissolved in 45-55 g of warm boiled water and then put into a water trough so that the female rats can take the feed freely, and the female rats are fed twice every day. By means of the method, the littering number of the female rats is large, the litter size is large, and the neonatal rats are large in weight, good in resistance, not prone to illness and high in survival rate.

The present invention relates to the field of biomedicines and particularly to a medicinal composition and an application thereof. The medicinal composition significantly reduces myocardial cell damage caused by doxorubicin and improves cell viability in SD neonatal rat myocardial cell injury tests; and after detecting mitochondrial ATPase activity of cardiomyocytes after combined administration,the combined application of caffeic acid with doxorubicin can enhance activity of Na<+>-K<+>-ATPase and Ca<2+>-ATPase in the cardiomyocytes. The combined administration of the caffeic acid with doxorubicin can prevent and treat myocardial damage caused by the doxorubicin, enhances viability of myocardial cells after injury, reduces degree of myocardial cell injury and apoptosis rates, protects mitochondria of the myocardial cells, reduces side effects of the doxorubicin, and expands scopes of clinical applications of the doxorubicin.

The invention discloses Clerodane diterpenoid compounds shown in 1-6 structural formulas, a pharmaceutical composition taking the Clerodane diterpenoid compounds 1-6 as effective components, a preparation method thereof and application thereof in preparing myocardial protection drugs. Pharmacological activity tests show the the compounds 1-6 can obviously increase the survival rate of myocardial cells, and at the same time, the compounds 1-6 are found to have better protective effect on H2O2-induced primary myocardial cell injury in neonatal rats, and the protective effect shows concentrationdependence. The research results show the the compounds can be developed into drugs for myocardial cell protection, and can be used for preventing and treating cardiovascular diseases such as coronaryheart disease, angina pectoris, myocardial infarction, myocarditis, arrhythmia and the like caused by myocardial ischemia and hypoxia.

The invention relates to the field of stem cells and regenerative medicine, and particularly relates to neural crest cell culture fluid, a preparation method of neural crest mesenchymal stem cells and an application of the neural crest mesenchymal stem cells. Experimental results of the invention show that mesenchymal stem cells of the neural crest stem cell lineage, induced and differentiated by human totipotent stem cells, can obviously improve neurological dysfunctions, reduce an area of an ischemic region, inhibit immune reactions and promote endogenous repairs of brain tissues in a neonatal rat hypoxic-ischemic model, thereby proving that human neural crest-derived mesenchymal stem cells have effects of significantly improving the inflammatory reaction of neonates with ischemic and anoxic encephalopathy and effectively promoting the neural function recovery. The invention also provides a test index for identifying the activity of the neural crest mesenchymal stem cells.

The invention provides a circular RNAcircTTC3 overexpression adeno-associated virus vector, an adeno-associated virus and application of the adeno-associated virus, and belongs to the technical field of biological medicine. The cyclic RNA-circTTC3 delivered by using the AAV9 virus vector can be used for preparing a medicine for preventing and / or treating myocardial ischemia-reperfusion injury (IRI). The gene sequence capable of transcribing the circular RNA-circTTC3 is delivered to the heart tissue based on the AAV virus, and is stably and highly expressed in the heart tissue, the circTTC3 is up-regulated to generate a heart protection effect, and the myocardial infarction area is effectively reduced, so that the effect of protecting myocardial infarction is achieved. According to the application disclosed by the invention, circTTC3 is over-expressed in primary myocardial cells of newborn rats, and the circTTC3 is found to be capable of remarkably promoting proliferation and growth of the primary myocardial cells of the newborn rats.