A kind of medical fibre Sigmoidoscope and preparation method thereofTechnical field
The invention belongs to area of medical devices, specifically, it is related to a kind of medical fibre Sigmoidoscope and preparation method thereof.
Background technology
Fibro-colonoscope is one kind of Sigmoidoscope.It is made up of optical glass fibre and interior mirror body and annex, can be to total colectomyLine-of-sighted observation.Suitable for checking that colon is suspected to have the lesions such as polyp, tumour, ulcer, inflammation, unknown cause hemorrhagic focus.Match somebody with somebodyX-ray clysis with barium or barium gas double contrast radiograph are closed, the diagnostic level of colonic pathological change can be improved.The major complications of inspection are enterobrosis.Fibro-colonoscope be applicable：Make the inspections and examinations of rectum, caecum, the colon ascendens, transverse colon, sigmoid colon, colon, and ileocecus etc.Treatment, due to can directly visually see the lesion of each part mentioned above, so as to diagnose correct, reliable.There is endoscope live body to takeModel machine structure.
Because in checking process, fibro-colonoscope enters rectum by anus, until large intestine, medical worker's observation colon andThe inner case of large intestine, thus the various bacteriums during fibro-colonoscope inevitably contacts enteron aisle in diagnoses and treatment and affected partPathological tissues, to pipeline cause infection, therefore be necessary to carry out it after inspection is finished targetedly thoroughly cleaning, with ensureThe clean and safe of each diagnoses and treatment, it is to avoid cross-infection occurs.Current cleaning method is that fibro-colonoscope is put into cleaningCleaning and sterilizing in pond or washing and disinfecting machine, but mirror body internal corners still there may be remnants patient's body in pathogen simultaneouslyThoroughly killing action can not be played.
A kind of detergent of fibro-colonoscope is disclosed in the A of CN 104498228, by the raw material system of following weightIt is standby to form：78 parts of water, four (3,5- di-t-butyl -4- hydroxyls) 25-27 parts of benzenpropanoic acid pentaerythritol esters, 0,0- dimethyl-S- (firstBase carbamoyl methyl) phosphorodithioate 20-22 parts, 26-28 parts of 3- (2,4_ 3,5-dimethylphenyl) -2- acrylic acid, 2,2- bis-Methyl isophthalic acid-propyl alcohol 16-18 parts, 2,2_ 12-15 parts of dimethyl -1- propyl alcohol diisooctyl succinate sodium sulfonates, the tertiary fourth oxygen of (S)-N-6-8 parts of carbonyl -2- benzene glycinols, 8-10 parts of 4- acetylaminohydroxyphenylarsonic acid 2 hydroxybenzoic acids, 1- (4- fluorophenyls) -4- [4- (4- chlorobenzenesBase) the light base -1- piperidyls of -4-] 4-6 parts of -1- butanone.Although the detergent is going in stain and sterilization on both side to ensure fiber colonThe cleaning effect of mirror, but remain the bad defect of bactericidal effect.
Antibacterial peptide is the small polypeptide that a class has very strong bactericidal effect to bacterium, since the eighties in last century, antibacterial peptideOne of constituent as the biological innate immunity start behave find and recognize constantly have new antibacterial peptide thereafter eachKind biology in find and be considered as anti-infective therapy future drugs.It has carrying positive charge, few resistance, fungicidal spectrumExtensively, species it is various, to the low feature of mammalian cell toxicity.The mechanism of action of antibacterial peptide is different from traditional antibiotic, diseaseOpportunistic pathogen is difficult to produce drug resistance to it, and some antibacterial peptides have very strong bactericidal activity to it, therefore is expected to be killed using antibacterial peptideThe infection of bacterium particularly pathogenic entero becteria.And the antibacterial peptide is used for the coating of the skin covering of the surface of fibro-colonoscope, you can realizeThe effect of the surface disinfection of fibro-colonoscope.
The content of the invention
A kind of defect that the purpose of the present invention exists aiming at prior art, there is provided simple structure, it is reasonable in design, preventThe cross-infection of remaining pathogenic bacteria during fibercolonscopy, prevents fibro-colonoscope from carrying the cross-infection of germ.
An object of the present invention is to provide a kind of anti-infective and suppression bacterium formation the carrying sustained-release antimicrobial peptide of strength to applyLayer medical fibre Sigmoidoscope.
The second object of the present invention is to provide a kind of manufacture method of carrying sustained-release antimicrobial peptide coating medical fibre Sigmoidoscope.
What the present invention was realized in：A kind of carrying sustained-release antimicrobial peptide coating medical fibre Sigmoidoscope, including fibro-colonoscopeBody, it is characterised in that：The surfaces externally and internally of the fibro-colonoscope is coated with antibacterial peptide slow release layer, the antibacterial peptide slow release layerTo be combined with antibacterial peptide polymer coating.
Above-mentioned antibiotic slow release layer thickness is 5-30 μm.
One of technical scheme of the invention provides a kind of preparation method of fibro-colonoscope antibacterial surface coating of peptides, willThe body of fibro-colonoscope immerses the aqueous solution of polyethyleneimine, and the concentration of polyethyleneimine is 5-10mg/ml in the aqueous solution,Soaked 40 minutes containing reciprocating vibration in 0.14mol/LNaCl, polymer base coat can be formed in the surfaces externally and internally of fibro-colonoscope；SoBe placed in that 100mL mass percent concentrations are 0.01% afterwards 4, react 5 hours in the methanol solution of 4 '-Diphenyl disulfide alcohol, insteadFibro-colonoscope after answering is vacuum dried after fully being embathed through methyl alcohol, with antibacterial peptide respectively in horseradish peroxidase and peroxidatingThe lower reaction of hydrogen effect, obtains the fibro-colonoscope of surface bond antibacterial polypeptide.When fibro-colonoscope is contacted with bacterial cell, pipeThe disulfide bonds in sublist face, reduction release polypeptide, play bactericidal action.
Described antibacterial peptide is that applicant's early stage screens acquisition, and its sequence is as follows：
KJT1：NTIWIPRRPHWQYWSQHWNTSPMTGLR
KJT2：QFFLQCGGGWVCSAHAWWMTRWWSQQR；
KJT3：WKHWGTIILNYEFRMNIAECFSGHDS；
KJT4：RLTWYPIYPMCHWCASAMYQYHHWLHQ；
KJT5：LSQVPGFIPMTHTRLQWLPVGPDQTRS；
KJT6：DRQHAATAQSLSLQYARFLVSWEQNNQ；
KJT7：RIEYHHNQYVTDVCSWQHKWDERGPWP；
The beneficial effects of the present invention are：Fibro-colonoscope with antibacterial peptide coating prepared by this method, with preventingFibro-colonoscope carries the germ in patient's body after use in vivo, and then causes other cross-patient contaminations to cause inflammation to be sent outRaw risk, with great application prospect and important social value value.Sterilization can be saved with the popularization of large areaCost, and reduce medical-risk.
Specific embodiment
The preparation of the fibro-colonoscope coat of embodiment 1
Fibro-colonoscope is cleaned by ultrasonic 5 minutes, 10 points in distilled water, acetone, 70% ethanol, ultra-pure water successively respectivelyClock, 10 minutes, after 5 minutes, then with ethane peroxide suffocating sterilization process after, immerse polyethyleneimine the aqueous solution, the aqueous solutionThe concentration of middle polyethyleneimine is 10mg/ml, and reciprocating vibration soaks 40 minutes in NaCl containing 0.14mol/L, can be in gastroscope pipeSurfaces externally and internally formed polymer base coat；Be subsequently placed in that 100mL mass percent concentrations are 0.01% 4,4 '-Diphenyl disulfideReacted 5 hours in the methanol solution of alcohol, reacted pipe is vacuum dried after fully being embathed through methyl alcohol, exists with KJT-1 antibacterial peptidesHorseradish peroxidase and the lower reaction of hydrogen peroxide effect, obtain the fibro-colonoscope of surface bond antibacterial polypeptide, are named asKJT-1 fibro-colonoscopes.
According to the method described above, KJT-2~7 fibro-colonoscope is prepared respectively.
The compliance test result of the antibacterial peptide of embodiment 2
It is 2mg/ml that KJT-1~7 polypeptide distilled water is dissolved into concentration, and antibacterial activity is carried out using agar diffusion methodDetection.The bacterial strain used in agar diffusion method is Listeria monocytogenes, vibrio parahemolyticus, campylobacter jejuni, smallYersinia enterocolitica, staphylococcus aureus, clostridium botulinum, C.perfringens, proteus, brucella, intestinesEnterohemorrhagic E.coli 0157:The combination of H7, Yersinia ruckeri and comma bacillus.After strain culturing, 50 microlitres of bacterium solutions are taken respectivelyCoat LB solid mediums.After standing band bacterium solution absorption, the polypeptide solution that concentration is 200ppm is separately added into.37 DEG C of cultures 1After hour, the presence or absence of inhibition zone and size are observed.The results are shown in Table 1.
The polypeptide Analysis of Antimicrobial Activity result of table 1
7 described polypeptides all have larger inhibition zone.The effect of the suppression pathogenic entero becteria having had in a short timeReally.
The hemolytic activity of the antibacterial peptide of embodiment 3
Collection people or the new blood lmL of rabbit, are dissolved into PBS solution after anticoagulant heparin, I000g centrifugation 5min, receiveCollection red blood cell；Washed with PBS 3 times, then it is resuspended with I0ml PBS；Take 100 μ L red cell suspensions and 100 μ L PBS dissolvings notIt is well mixed with the antibacterial peptide solution of concentration, the constant-temperature incubation Ih in 37 DEG C of incubators；Taken out after Ih, 4 DEG C of 1000g centrifugations5min；Take out supernatant ELIASA light-metering absorption value at 540nm；Every group is averaged, and comparative analysis.Wherein 100 μ1PBS is used as negative control；100 μ 10.2%Tritonx-100 are used as positive control.Result finds that 7 antibacterial peptides exist respectivelyThere is no the haemolysis imagination under the concentration of 2000 micrograms/ml, with great application space.
The antibiotic property detection of fibro-colonoscope of the embodiment 4 containing polypeptides coating
7 kinds of fibro-colonoscopes that embodiment 1 is prepared and the fibro-colonoscope without polypeptides coating, give respectivelyWith EHEC 0157:H7 positive intestines problem patient uses, after use, by conventional cleaning and sterilization sideFormula is cleaned.Afterwards, in aseptic environment, fibro-colonoscope surfaces externally and internally is respectively placed in PBS water environments and is shaken, is rushed30min is washed, then by bacteriological filtration membrane filtration water body, bacteriological filtration film surface is rinsed with culture medium, 37 degrees Celsius of culture 24h, afterwardsDetection finds, using not detecting EHEC 0157 in 7 kinds of gastroscopes that embodiment 1 is prepared:H7 is thinThe presence of bacterium, without detecting EHEC 0157 in the fibro-colonoscope of polypeptides coating:The presence of H7, it is total to causeGerm bacterium number amounts to 35.This also illustrates that polypeptides coating has preferably suppress what bacterium remained in general fibre SigmoidoscopeFunction.
Coating prepared by other several bacterium uses other several pathogens：Yersinia ruckeri and comma bacillus carry out above-mentioned sameThe inspection of sample, as a result shows the presence for not detecting pathogen in the fibro-colonoscope containing antibacterial peptide coating.
It is above specific embodiment of the invention, its role is to be further described in detail to present disclosure,Reader is more readily understood, but is not constituted to protection of the presently claimed invention.
Sequence table
The ＞ Nanchang Qing Nang traditional Chinese medicines Co., Ltds of ＜ 110
A kind of medical fibre Sigmoidoscopes of the ＞ of ＜ 120 and preparation method thereof
〈210〉1
〈211〉27
〈212〉PRT
〈400〉KJT1
NTIWIPRRPHWQYWSQHWNTSPMTGLR
〈210〉2
〈211〉27
〈212〉PRT
〈400〉KJT2
QFFLQCGGGWVCSAHAWWMTRWWSQQR；
〈210〉3
〈211〉26
〈212〉PRT
〈400〉KJT3
WKHWGTIILNYEFRMNIAECFSGHDS；
〈210〉4
〈211〉27
〈212〉PRT
〈400〉KJT4
RLTWYPIYPMCHWCASAMYQYHHWLHQ；
〈210〉5
〈211〉27
〈212〉PRT
〈400〉KJT5
LSQVPGFIPMTHTRLQWLPVGPDQTRS；
〈210〉6
〈211〉27
〈212〉PRT
〈400〉KJT6
DRQHAATAQSLSLQYARFLVSWEQNNQ；
〈210〉7
〈211〉27
〈212〉PRT
〈400〉KJT7
RIEYHHNQYVTDVCSWQHKWDERGPWP；