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 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 invention belongs to the technical field of medicines and pharmacology and discloses an identification method and system for anoxia-reoxygenation injury of myocardial cells. Telomerase mitochondrion transposition is regulated and controlled, mitochondria damage can be alleviated, and myocardial energy supply can be improved, so that a myocardial cell protective effect is achieved. The identification method and system which are disclosed by the invention have the advantages that activity and distribution of telomerases are regulated through overexpression of Shp-2ShRNA interference plasmidsand mitochondrion-targeted hTERT at a primary culture neonatal rat myocardial cell level, and the mitochondria damage is analyzed and is also alleviated; and relation between the telomerase mitochondrion transposition and mitochondrial functions is clear, a possible mechanism of the telomerase mitochondrion transposition for alleviating the anoxia-reoxygenation injury of the myocardial cells is analyzed, and a basis is provided for a myocardial protection treatment strategy taking telomerase as a target.

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 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 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 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 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.