47 results about "Horseshoe crab" patented technology

An artificial breeding and culturing method for horseshoe crabs includes: mixing eggs of horseshoe crabs with bodily fluid containing sperms, and allowing the mixture to stand until fertilization; putting the fertilized eggs into filtered seawater to be incubated, and filling oxygen until the eggs are incubated into one-year-old horseshoe crabs; breeding the one-year-old horseshoe crabs at the temperature of from 28 DEG C to 30 DEG C until the crabs slough and develop into two-year-old horseshoe crabs; breeding the two-year-old horseshoe crabs at the temperature of from 20 DEG C to 28 DEG C, and using an air pump to fill oxygen; taking out the two-year-old horseshoe crabs and placing into another container containing bait regularly each day, changing the filtered seawater once a day untilthe crabs are bred into five-year-old crabs; breeding the horseshoe crabs which are five years old or of different ages at the temperature of from 20 DEG C to 28 DEG C, and filling oxygen; taking outthe crabs and breeding in another container containing bait regularly each day, and changing the filtered seawater once a day; and taking out the crabs in breeding and breeding in a container or cement pit with oyster fragments until the crabs are bred into adult horseshoe crabs.

Objects of the present invention are to provide a DNA fragment encoding a limulus-derived pro-clotting enzyme, a virus harboring the DNA fragment, a cell harboring the virus, a method of producing the pro-clotting enzyme by use of the cell, and means for assaying an endotoxin or (1→3)-β-D-glucan employing the enzyme, wherein these elements are capable of producing an endotoxin or (1→3)-β-D-glucan assay reagent of satisfactory quality, steadily, at low cost, and on a large scale. In the present invention, for example, a DNA fragment encoding a protein having an amino acid sequence defined by SEQ ID NO: 4 is selected as a nucleic acid fragment encoding a limulus-derived pro-clotting enzyme, and the corresponding recombinant pro-clotting enzyme. Use of the enzyme can provide a high-sensitivity method and kit for detecting (1→3)-β-D-glucan and an endotoxin, utilizing a cascade reaction system in a horseshoe crab lysate.

The invention relates to a method for largely and quickly extracting tachyplesin peptide. The method comprises the following steps: (1) collecting horseshoe crab blood cells for reservation, suspending the collecting horseshoe crab blood cells in Tris-HCl and NaCl buffer solution homogenate, performing centrifugation for removing supernatant liquor, and washing sedimentation with the same buffer solution twice; (2) depositing, adding acid liquor homogenate according to the ratio of solid to liquid of 1: (6.5-8), performing centrifugation for collecting supernatant liquor, and re-extracting the sedimentation for three times by using acid liquor to obtain acid extraction liquid; (3) adjusting the pH of the acid extraction liquid to 6.5 by using NaOH, performing centrifugation to remove sedimentation, and performing filtration by use a 0.1-0.22 [Mu]m film to obtain supernatant liquor; (4) performing boiling water bath processing on the supernatant liquor, performing centrifugation to remove sedimentation, collecting supernatant liquor, performing filtration by use the 0.1.22 [Mu]m film to obtain supernatant liquor; (5) performing processing on ultrafiltration membrane with the molecular weight cut off of 10 KD; (6) performing processing on ultrafiltration membrane with molecular weight cut off of 3 KD; (7) performing desalination processing; and (8) performing freeze drying on the obtained product to be made into dry powder. The tachyplesin peptide obtained by using the method is high in yield and purity; the process is simplified; the time is greatly shortened; the cost is greatly reduced; and the throughout is increased significantly.

The invention belongs to the field of traditional Chinese medicines (TCMs) and discloses a TCM composition and application thereof. The composition comprises the following TCMs in parts by weight: 0.11-97.0 parts of musk, 10.0-50.0 parts of gypsum, 0.05-10.0 parts of borneol, 5.0-15.0 parts of pseudo-ginseng, 4.5-9.0 parts of cuttlebone, 3.0-9.0 parts of silkworm cocoons, 3.0-6.0 parts of human urine sediment, 4.0-80.0 parts of calcined gypsum, 0.5-25.0 parts of pearls, 2.0-12.0 parts of frankincense, 2.0-12.0 parts of myrrh, 1.0-10.0 parts of sanguis draconis, 1.0-10.0 parts of rhizoma coptidis, 3.0-20.0 parts of horseshoe crab shells, 0.2-2.0 parts of dragon bones, 9.0-30.0 parts of oysters and 0.5-15.0 parts of carbonized human hair. The TCM composition has the functions of clearing away heat and toxic materials, activating blood circulation to dissipate blood stasis, carrying out sterilization to diminish inflammation, relieving pains, diminishing swelling and promoting granulation, inducing astringency and stopping slippage and astringing dampness and carrying out astringency. The TCM composition can be used for preparing the medicines for treating bedsore, ecthyma, scrofula sore, hemorrhoids, burns, scalds, tympanitis or diabetic foot.

The invention relates to a developing-process fungus 1,3-beta-D-glucan detection kit for human body fluid. The developing-process fungus 1,3-beta-D-glucan detection kit comprises a reaction main agent, a main agent compound solution, a sample treatment solution, heat-source-free water, a standard product and a quality control product, wherein the reaction main agent takes horseshoe crab blood cells as a main raw material and contains G factors, coagulase, coagulase zymogen and a polypeptide developing substrate; the polypeptide developing substrate is synthesized tripeptide or tetrapeptide with a Gly-Arg tail end connected with a PNA; the polypeptide developing substrate is subjected to enzyme digestion by adopting the coagulase; after the free paranitroaniline (PNA) is generated, a microplate reader is used for directly detecting so that a detection route is shortened and the cost is reduced; the microplate reader is used for carrying out a velocity-method enzyme kinetics detection method so that the sensitivity is relatively high when being compared with a nephelometry detection method; and the reaction main agent is not easily interfered by protein in a body fluid sample and medicines to generate non-specific turbidity, so that the probability of a false positive detection result is reduced and the detection accuracy is relatively high.

The invented testing method solves blindfold determination of important parameter-terminal hour in prior art, making selection of terminal hour possess rationality. Adopting photoelectric detecting instrument, the invention records, plots dynamic curve of reaction, and searches out optimal terminal hour by using computer. Then, end-point method is carried out for testing content of endotoxin in sample quantificationally by utilizing the said terminal hour. The method is applicable to horseshoe crab reagent in end-point method as well as reagents in dynamic turbidimetry or dynamic colorimetry.

The invention discloses an analog peptide molecule of antiendotoxin factor of Limulida to kill bacteria and treat endotoxin blood symptom, which applies computer molecular analog technique to design the following 6 groups of analog peptides antiendotoxin factor of Limulida based on GOR method with TALF functional area position as mould: RX1 NPT X2 KRLK X3 KYKGK X3 WCP as the first group, X4 INPT X2 KR X1 X4 X3 KYKGKFW as the second group, INPT X2 KRLKW X4 YKGR X3 X3 CP as the third group, PT X2 KR X1 KX3 KYKGKFWCP as the forth group, K X2 NPTVKR X1 K X3 RYKGKF as the fifth group, NPX5 VKR X1 K X3 RYKGKF as the sixth group, wherein six groups are synthesized into the peptide through solid phased peptide synthetic method by self-synthesizing and manual synthesizing, which produces the peptide with sterilizing and/or neutralizing LPS activity.

To provide a method for producing a horseshoe crab recombinant Factor C. The horseshoe crab recombinant Factor C is produced through expression thereof by use of mammalian cells such as CHO DG44 and HEK293 as host cells.

The invention discloses a salve for treating zoster, which includes components of marine horseshoe crab shells and tea oil. The salve for treating zoster adopts marine horseshoe crab shells as main medicine and tea oil as supplemented drugs, the combined use of them can rapidly alleviate neuralgia resulted from zoster and can effectively prevent infection contamination, to achieve the object of rapidly curing zoster.

The invention discloses a method for obtaining natural horseshoe crab fertilized eggs by building a pool bottom overhead, laying sand and conducting partition. The method is characterized by including the steps of firstly, selecting parent horseshoe crabs, wherein three or more pairs of parent horseshoe crabs are selected as the parents in the mating season each year; secondly, refitting a parent horseshoe crab pool indoors, wherein the pool bottom of a breeding pool is built overhead through a wood frame, a 60-mesh bolting-silk net is laid on the pool bottom, an oxygen pipe used for inflation is arranged in the overhead wood frame, and sand is laid on an egg laying area; thirdly, breeding the parent horseshoe crabs indoors, wherein the parent horseshoe crabs selected in the first step are placed in the parent horseshoe crab pool refitted in the second step, and indoor inflation breeding is conducted; fourthly, making the parent horseshoe crabs lay eggs and release semens, wherein the parent horseshoe crabs can lay eggs and release semens in 2 days to 10 days after entering the breeding season to form fertilized eggs; fifthly, sieving and washing the fertilized eggs; wherein after the fertilized eggs are hardened, the fertilized eggs and sand are dug out, elutriation is conducted through a screen with the diameter of 2 mm, all the sand is leaked out, and the fertilized eggs are left. Different from a method for obtaining the fertilized eggs by killing horseshoe crabs, the method has the advantages that a large number of high-quality fertilized eggs can be obtained, and the method is easy and convenient to operate, efficient, highly feasible and the like.

The present invention discloses a herpes zoster treatment Chinese patent medicine, which is prepared from the following raw materials, by weight: 18-29 parts of medicinal morinda root, 15-27 parts of tatarinow sweetflag rhizome, 9-17 parts of Chinese pulsatilla root, 10-19 parts of horseshoe crab carapace, 15-20 parts of sharpleaf uncaria stem with hooks, 19-27 parts of rheum, 15-19 parts of sapindus mukurossi, 12-19 parts of snake slough, and 10-16 parts of isatis tinctoris. The herpes zoster treatment Chinese patent medicine has the following advantages that: the medicine is prepared from Chinese herbs, has effects of no toxic-side effects, anti-inflammation, pain relieving, detoxification, sterilization, heat clearing and blood cooling, has characteristics of adequate raw materials, simple production process, low price and convenient use, and is the external application specially good effect medicine for treating skin herpes zoster, and clinical case treatment results verify that an effective rate is 100%, a cure rate is 91.7%, and no pigmentation and sequel is generated after healing. In addition, the herpes zoster treatment Chinese patent medicine provides a certain treatment effect for other skin herpes and papule.

The invention discloses a method for preparing kynurenine from horseshoe crab tails.The method comprises the following steps of preparation of dry horseshoe crab tail powder, preparation of horseshoe crab tail crude extract, primary separation and purification of the horseshoe crab tail crude extract, separation and purification of a component A8 and separation and purification of components A8-24, wherein separation and purification of the components A8-24 comprise the steps that an appropriate amount of components A8-24 are taken, methyl alcohol is added for dissolution, and normal phase column separation is conducted; 50 mg of samples are taken and stirred with 150 mg of silica gel powder, 5 g of silica gel is saturated with petroleum ether and then subjected to column packing, after the the samples are packed in a column, elution is conducted with petroleum ether and acetone, when the ratio of petroleum ether to acetone is 1:10, fractions are collected, the number of the fractions is 60, the fractions from the 25 tube to the 40 tube are merged, after the the fractions are merged, vacuum concentration is conducted, and after solvent is dried out, kynurenine is obtained.According to the method for preparing kynurenine from the horseshoe crab tails, kynurenine is separated from the horseshoe crab tails for the first time, and kynurenine serving as the active ingredient of the horseshoe crab tails has great significance in secondary development of horseshoe crab tails.

The invention discloses a restoration culture method for an injured Chinese horseshoe crab. The method comprises the following steps: cleaning the internal wall and bottom of a pool with an garlic detergent, sprinkling an oxalic acid solution for disinfection, and flushing the repair pool with fresh water on next day until no oxalic acid solution remains; laying a sandy bottom of 20cm-30cm, carrying out soaking disinfection for 30min with 10ppm potassium permanganate, and flushing the sandy bottom with fresh water until no potassium permanganate remains; subjecting the repair pool to sun curing for 24 hours, and adding fresh seawater subjected to sand filtration, dark and ultraviolet light disinfecting treatment; uniformly laying gas pipes to continuously inflate, so as to keep dissolved oxygen to be 5mg/L or more; rinsing out the injured Chinese horseshoe crab with the seawater in the repair pool, and carrying out disinfecting for 15min; placing the treated injured Chinese horseshoe crab into the repair pool, and carrying out culture with running water; lowering the water level to 15cm to 20cm every morning; feeding fresh waste fish or oyster meat in the afternoon of first severaldays, and cleaning remaining remnant bait after the injured Chinese horseshoe crab finishes food intake; adjusting a feeding amount at a later stage according to food intake conditions; and carryingout culture until rapid body contraction reduction is achieved through touch in water and the food intake conditions are stable.

The invention discloses new application of tachyplesin peptides. The tachyplesin peptides can be used for preparing medicine and healthcare products for preventing and treating pulmonary fibrosis, used as the effective component of the medicine and healthcare products for preventing and treating pulmonary fibrosis, used for the prodrug modification of the medicine for preventing and treating pulmonary fibrosis or used along with the preventing and treating pulmonary fibrosis to treat the pulmonary fibrosis. The tachyplesin peptides are prepared by waste materials of horseshoe crab blood cell or horseshoe crab reagent production and can evidently inhibit the formation of the pulmonary fibrosis.

The buffering liquid for limulus test for detecting bacterian endotoxin is Tris-HCl buffering liquid containing phenol red in 0.005-0.1 wt% concentration. The preparation process includes weighing phenol red and Tris salt and stoving at 150-200 deg.c for 1-5 hr; adding no-endotoxin water and regulating pH to required value with concentrated hydrochloric acid; high pressure sterilization and no-bacteria preservation or freezing preservation. The buffering liquid is used in diluting tested sample and regulating and indicating sample pH value. The buffering liquid for limulus test for detecting bacterian endotoxin has no influence on the endotoxin in the detection of two kinds of endotoxin and coincides with the requirement for limulus test.

The invention discloses a horseshoe crab antiviral combined extract, a preparation method and an application of the horseshoe crab antiviral combined extract to disinfection products. Two extracts with antiviral effects are respectively obtained from horseshoe crab serum and horseshoe crab blood cells, and the extracts have more obvious inhibition effects on viruses when being combined for use. The novel coronavirus resisting activity of a spray dressing formed by combining the two extracts reaches up to 99.51%, and the influenza A virus resisting activity is 42.58%. The two extracts can be extracted from waste materials obtained from horseshoe crab reagent production, so that the production waste materials are recycled, and biological resources can also be fully utilized while the production cost is reduced. The two antiviral extracts can be prepared to obtain dressings for nasal cavity and skin disinfection. Compared with traditional chemical disinfection products, the two antiviral extracts are safer, milder and more effective, have no toxic and side effects, and can avoid drug resistance and allergy phenomena of conventional disinfection products.