A kind of medical fibre Sigmoidoscope and preparation method thereofTechnical field
The invention belongs to area of medical devices, specifically, are related to a kind of medical fibre Sigmoidoscope and preparation method thereof.
Background technique
Fibro-colonoscope is one kind of Sigmoidoscope.It is made of optical glass fibre and interior mirror body and attachment, it can be to total colectomyIt is line-of-sighted observation.Suitable for checking that colon is suspected to have the lesions such as polyp, tumour, ulcer, inflammation, unknown cause hemorrhagic focus.MatchX-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 is applicable in：Make rectum, caecum, colon ascendens, transverse colon, sigmoid colon, the inspections and examinations of colon and ileocecus etc.Treatment, due to can directly visually see the lesion of each part mentioned above, so that diagnosis is correct, reliable.Endoscope takes with living bodyModel machine structure.
Since in checking process, fibro-colonoscope enters rectum by anus, until large intestine, medical worker observe colon andThe inner case of large intestine, thus fibro-colonoscope inevitably contacts various bacteriums and the affected part in enteron aisle in diagnoses and treatmentPathological tissues, pipeline is caused to infect, therefore it is necessary to targetedly thoroughly clean it after inspection, to guaranteeThe clean and safe of each diagnoses and treatment avoids that 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 the pathogens of remaining patient's body simultaneouslyThorough killing effect can not be played.
A kind of detergent of fibro-colonoscope is disclosed in 104498228 A of CN, by the raw material system of following weightIt is standby to form：78 parts of water, four 25-27 parts of benzenpropanoic acid pentaerythritol ester of (3,5- di-t-butyl -4- hydroxyls), 0,0- dimethyl-S- (firstBase carbamoyl methyl) 20-22 parts of phosphorodithioate, 26-28 parts of -2- acrylic acid of 3- (2,4_ 3,5-dimethylphenyl), 2,2- bis-Methyl-1-16-18 parts of propyl alcohol, 2,2_ 12-15 parts of propyl alcohol diisooctyl succinate sodium sulfonate of dimethyl-1-, the tertiary fourth oxygen of (S)-N-6-8 parts of benzene glycinol of carbonyl -2-, 8-10 parts of 4- acetylaminohydroxyphenylarsonic acid 2 hydroxybenzoic acid, 1- (4- fluorophenyl) -4- [4- (4- chlorobenzeneBase) the light base -1- piperidyl of -4-] 4-6 parts of -1- butanone.Although the detergent is removing stain and sterilization upper guarantee fiber colon on both sideThe cleaning effect of mirror, but remain the bad defect of bactericidal effect.
Antibacterial peptide is a kind of small polypeptide to bacterium with very strong bactericidal effect, 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 each thereafterThe future drugs of anti-infective therapy are found and are considered as in kind biology.It, which has, carries positive charge, few drug resistance, fungicidal spectrumExtensively, the features such as many kinds of, low to mammalian cell toxicity.The mechanism of action of antibacterial peptide is different from traditional antibiotic, diseaseOpportunistic pathogen is not easy to generate it drug resistance, and some antibacterial peptides have very strong bactericidal activity to it, therefore is expected to kill using antibacterial peptideThe infection of bacterium especially pathogenic entero becteria.And the antibacterial peptide is used for the coating of the skin covering of the surface of fibro-colonoscope, it can be realizedThe effect of the surface disinfection of fibro-colonoscope.
Summary of the invention
The purpose of the present invention is to defect of the existing technology, provide that a kind of structure is simple, and design rationally, preventsThe cross-infection of remaining pathogenic bacteria, prevents fibro-colonoscope from carrying the cross-infection of germ during fibercolonscopy.
One of the objects of the present invention is to provide the carrying sustained-release antimicrobial peptide paintings that a kind of strength is anti-infective and bacterium is inhibited to be formedLayer medical fibre Sigmoidoscope.
The second object of the present invention is to provide a kind of manufacturing method of carrying sustained-release antimicrobial peptide coating medical fibre Sigmoidoscope.
The invention is realized in this way：A kind of carrying sustained-release antimicrobial peptide coating medical fibre Sigmoidoscope, including fibro-colonoscopeOntology, 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 is with a thickness of 5-30 μm.
One of technical solution of the invention provides a kind of preparation method of fibro-colonoscope antibacterial surface coating of peptides, willThe ontology of fibro-colonoscope immerses the aqueous solution of polyethyleneimine, and the concentration of polyethyleneimine is 5-10mg/ml in the aqueous solution,It is impregnated 40 minutes containing reciprocating vibration in 0.14mol/LNaCl, polymer base coat can be formed in the surfaces externally and internally of fibro-colonoscope；SoIt is placed in the methanol solution for 4, the 4 '-Diphenyl disulfide alcohol that 100mL mass percent concentration is 0.01% and reacts 5 hours, insteadFibro-colonoscope after answering is dried in vacuo after methanol sufficiently embathes, 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 and bacterial cell contact, pipeThe disulfide bonds in sublist face, reduction release polypeptide, play bactericidal effect.
The antibacterial peptide is to screen acquisition applicant's early period, and 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：The fibro-colonoscope with antibacterial peptide coating of this method preparation, having preventsFibro-colonoscope carries the germ of patient's body after use in vivo, and then causes other cross-patient contaminations that inflammation is caused to be sent outRaw risk, with great application prospect and important social value value.Disinfection can be saved with the popularization of large areaCost, and reduce medical-risk.
Specific embodiment
The preparation of 1 fibro-colonoscope coat of embodiment
Fibro-colonoscope is successively cleaned by ultrasonic 5 minutes, 10 points in distilled water, acetone, 70% ethyl alcohol, ultrapure water respectivelyClock, 10 minutes, after five minutes, then with ethane peroxide suffocating sterilization handle after, immerse the aqueous solution of polyethyleneimine, the aqueous solutionThe concentration of middle polyethyleneimine is 10mg/ml, and reciprocating vibration impregnates 40 minutes in NaCl containing 0.14mol/L, can be in gastroscope pipeSurfaces externally and internally formed polymer base coat；It is subsequently placed in 4, the 4 '-Diphenyl disulfides that 100mL mass percent concentration is 0.01%It is reacted 5 hours in the methanol solution of alcohol, the pipe after reaction is dried in vacuo after methanol sufficiently embathes, and is existed with KJT-1 antibacterial peptideHorseradish peroxidase and the lower reaction of hydrogen peroxide effect, obtain the fibro-colonoscope of surface bond antibacterial polypeptide, are named asKJT-1 fibro-colonoscope.
According to the method described above, the fibro-colonoscope of KJT-2~7 is prepared respectively.
The compliance test result of 2 antibacterial peptide of embodiment
It is 2mg/ml that the polypeptide of KJT-1~7, which is dissolved into concentration with distilled water, carries out antibacterial activity using agar diffusion methodDetection.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:H7, Yersinia ruckeri and the combination of comma bacillus.After strain culturing, 50 microlitres of bacterium solutions are taken respectivelyIt is coated on LB solid medium.After standing band bacterium solution absorption, it is separately added into the polypeptide solution that concentration is 200ppm.37 DEG C of cultures 1After hour, the presence or absence of observation inhibition zone and size.It the results are shown in Table 1.
1 polypeptide Analysis of Antimicrobial Activity result of table
7 polypeptides all have biggish inhibition zone.The effect for the inhibition pathogenic entero becteria having had in a short timeFruit.
The hemolytic activity of 3 antibacterial peptide of embodiment
The new blood lmL of people or rabbit is acquired, is dissolved into PBS solution after anticoagulant heparin, I000g is centrifuged 5min, receivesCollect red blood cell；It is washed 3 times with PBS, then is resuspended with I0ml PBS；100 μ L red cell suspensions and 100 μ L PBS is taken to dissolve notAntibacterial peptide solution with concentration is uniformly mixed, the constant-temperature incubation Ih in 37 DEG C of incubators；It is taken out after Ih, 4 DEG C of 1000g centrifugations5min；It takes out supernatant microplate reader and surveys absorbance value at 540nm；Every group is averaged, and comparative analysis.Wherein 100 μ1PBS is as negative control；100 μ 10.2%Tritonx-100 are as positive control.As a result, it has been found that 7 antibacterial peptides exist respectivelyThere is no the haemolysis imaginations under 2000 micrograms/ml concentration, have great application space.
The antibiotic property of fibro-colonoscope of the embodiment 4 containing polypeptides coating detects
7 kinds of fibro-colonoscopes that embodiment 1 is prepared and the not fibro-colonoscope of polypeptides coating, give respectivelyWith enterohemorrhagic escherichia coli 0157:The intestines problem patient of the H7 positive uses, and after use, passes through conventional cleaning and sterilization sideFormula is cleaned.Later, in sterile environment, fibro-colonoscope surfaces externally and internally is respectively placed in PBS water environment and shakes, rush30min is washed, water body is then filtered by bacteriological filtration film, bacteriological filtration film surface is rinsed with culture medium, 37 degrees Celsius are cultivated for 24 hours, laterDetection is found, does not detect enterohemorrhagic escherichia coli 0157 in 7 kinds of gastroscopes being prepared using embodiment 1:H7 is thinThe presence of bacterium, without detecting enterohemorrhagic escherichia coli 0157 in the fibro-colonoscope of polypeptides coating:The presence of H7, it is total to causeGerm bacterium number is 35 total.This also illustrates that polypeptides coating has preferable inhibition bacterium remaining in general fibre SigmoidoscopeFunction.
The coating of other several bacterium preparations uses other several pathogens：Yersinia ruckeri and comma bacillus carry out above-mentioned sameThe inspection of sample does not detect the presence of pathogen in the fibro-colonoscope as the result is shown containing antibacterial peptide coating.
The above are specific embodiments of the present invention, its role is to be further described in detail to the contents of the present invention,It is more readily understood reader, but is not constituted to protection of the presently claimed invention.
Sequence table
110 Nanchang > Qing Nang traditional Chinese medicine Co., Ltd of <
A kind of medical fibre Sigmoidoscope of 120 > of < 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；