CN110278919A - A method of building vermiform mite causes acne rosacea sample skin lesion animal model - Google Patents
A method of building vermiform mite causes acne rosacea sample skin lesion animal model Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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Abstract
The present invention discloses a kind of method that building vermiform mite causes acne rosacea sample skin lesion animal model, including by facial vermiform mite it is positive for mite person's clearing skin, it wipes dry, squeeze the scraping wing of nose or facial sebum secretion object, by cleaning secretion after extruding, vermiform mite protozoon suspension is prepared, and by vermiform mite protozoon suspension intracutaneous injection in the Japanese big test block ear rabbit back 4cm × 4cm.The present invention is for existing rabbit to the reactive very little of people's vermiform mite natural infection approach, lack the problem of vermiform mite causes acne rosacea sample skin lesion animal model, using Japanese big ear rabbit strain simultaneously in intradermal vaccination vermiform mite protozoon, it constructs vermiform mite and causes acne rosacea sample skin lesion animal model, the big ear rabbit damage of Japan is observed that similar erythema, papule skin lesion clinical manifestation and Histopathologic appearance with the acne rosacea of mankind vermiform mite positive height, meets the requirement that acne rosacea sample skin lesion animal model is caused as vermiform mite.
Description
Technical field
The present invention relates to a kind of animal model technical fields, and in particular to a kind of building vermiform mite cause acne rosacea sample skin lesion
The method of animal model.
Background technique
Acne rosacea is a kind of a kind of common chronic inflammation for involving patient facial region's skin heart and pilosebaceous follicle
Skin disease, clinical manifestation be patient's Middle face based on transient or erythema perstans, telangiectasis, papule, warts, one
A little longer patients of the course of disease are due to hyperblastosis it may also occur that the patch of kermesinus even pachydermatosis.Currently, the morbidity of acne rosacea
Rate increases year by year, and treatment lacks specific drug.Since the disease damages patient facial region's beauty, thus and life matter physically and mentally healthy to patient
Amount is all influenced, some patients will appear depression, feel oneself inferior, and cowardice is unsociable and eccentric, and mental burden weight loses the letter of life
The heart.Acne rosacea is increasingly becoming the hot spot of medical field research, at present it is generally acknowledged that the generation and heredity, Cellular inflammatory of acne rosacea
The factor, nervus vasculairs insufficiency of accommodation etc. are related.In recent years studies have shown that the morbidity of vermiform mite and acne rosacea has substantial connection.
Vermiform mite is commonly called as demodex folliculorum, parasitizes only two kinds of human body, i.e. demodex folliculorum and demodex brevis.Vermiform mite is posted
It is born in the hair follicle and sebaceous glands of people, has the characteristics that high infectious, low pathogenicity and opportunistic.It has been found that vermiform mite can
See following skin obstacle: with or not with skin lesions such as the facial itch of erythema, telangiectasis.It is multinomial both at home and abroad to grind
Study carefully and shows that the vermiform mite recall rate of acne rosacea patient facial region is apparently higher than normal person.China Zhu Xiao swallow etc. uses adhesive tape
It pastes method and vermiform mite counting is carried out to the skin sample of 164 student enrollment, find the vermiform mite of acne rosacea patient skin
Quantity is significantly more than normal population, also demonstrates the facial skin lesion such as vermiform mite and acne rosacea and has and definitely contacts.It is related at present
Vermiform mite participates in the mechanism of the morbidity of acne rosacea, mainly there is the following aspects:
1. the barrier function of vermiform mite damage skin
The destruction of the barrier function of skin is the initiating agent of a variety of maxillofacial diseases.Foreign countries, which publish an article, reports acne rosacea
Lesions of patients finds vermiform mite, and mention after its skin lesion tissue of serial section in the structures such as its pathologic granuloma
In serial section, there is vermiform mite particle in Chang Faxian pathological tissue, therefore speculates that vermiform mite is tardy because tissue can be caused to occur
Type hypersensitivity and cause a disease.In the keratinocyte at acne rosacea skin lesion, the part residuum of vermiform mite can induce TLR2
Overexpression and activation and KLK5 expression increase.KLK5, which has, makes cutin desmosome that degradation, Cell tracking hair occur
The effect of raw dissociation, thus the barrier function for making keratoderma that cannot bring into normal play because losing integrality；KLK5 can also swash
The NF- κ B access of PAR2 and downstream living, the expression of the inflammatory factors such as up-regulation IL-8, TSLP and TNF；KLK5 believes in PAR2 simultaneously
It plays a role during the expression of number access, hinders lamellar body release, keep stratum corneum penetration barrier function good from obtaining
Reparation.There is research to identify the presence of heat shock protein and lipoprotein from the skin lesion of acne rosacea, both albumen are all
The agonist of TLR2.Under the induction of vermiform mite etc., skin keratin forms TLR2 quantity expressed by cell and activity increases,
Epidermis KLK5 activation causes desmosome between keratinocyte to degrade, inhibit lamellar body release and lead to the poly- of inflammatory factor
Collection, to compromise physical skin barrier and immunization barrier function.These all illustrate that vermiform mite can in acne rosacea patient's body
It can cause, have activated a series of immune response.
2. vermiform mite influences skin heart, nerve modulation
The different subtype of acne rosacea at least 500 genes are different from normal skin, these abnormal genes are related on kidney
Adrenoceptor, protease etc., that is, be related to regulation of blood vessels；It is related to transient receptor potential, capsaicin receptor 1 etc., that is, generates mind
Through source property inflammation.Capsaicin receptor belongs to a member of transient receptor potential family, after it is activated by vermiform mite, a releasable system
Column inflammatory mediator leads to the generation of pain and neuroinflammatory conditions, and acne rosacea patient is caused to generate the discomforts such as shouting pain, burn feeling,
These inflammatory mediators include calcitonin gene-related peptide and Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met-NH2 etc..Also correlative study shows that acne rosacea patient may deposit
The relevant gene mutation of receptor is adjusted in neural blood vessel or there are tumor susceptibility genes, under the induction of many factors such as vermiform mite,
Neuropeptide is discharged by transient receptor potential, causes characteristic flush, oedema even persistent erythema.
From the point of view of research both at home and abroad at present, the research of vermiform mite becomes the hot topic of Human Parasite Science and dermatological field
Project.Therefore, the animal model for establishing acne rosacea caused by vermiform mite carries out the research of acne rosacea caused by vermiform mite in a deep going way,
It understands fully the cause of disease and pathogenesis, develops and cure acne rosacea drug, for mitigation patients ' psychological burden, improve the quality of living, it will
Generate positive effect.
Summary of the invention
In view of above-mentioned, acne rosacea disease incidence height, in order to study the pathogenesis of acne rosacea caused by vermiform mite, the present invention
Be designed to provide it is a kind of building vermiform mite cause acne rosacea sample skin lesion animal model method, can be succeeded using this method
Construct the big ear model of rabbit of Japan of acne rosacea sample skin lesion caused by vermiform mite.
The purpose of the present invention is be achieved through the following technical solutions:
A method of building vermiform mite causes acne rosacea sample skin lesion animal model, the steps include:
A. the preparation of vermiform mite protozoon suspension is cleaned
1. for mite person: the 9th 4 〇 hospital dermatology department out-patient of joint logistics system army, the Chinese People's Liberation Army is diagnosed as rose
Acne, and acarid microscopy is positive；
2. preparation method: for mite person's warm water cleaning skin, wiping dry, then squeeze the scraping wing of nose or facial sebum point using acari taking device
2% dish washing liquid of drop is added in the sebum secretion object of acquisition, mixes to homogeneous latex emulsion shape, and homogeneous latex emulsion shape is shifted for secretion
Into the teat glass of 5ml, 1 × PBS that 2ml is added is diluted, and is blown and beaten and is mixed with ear washing bulb, is placed in 4 DEG C of refrigerators and is stood certainly
So precipitating 2h, after abandoning the supernatant in teat glass, 1 × PBS that 2ml is added again is diluted and precipitates, and abandons supernatant
Afterwards, repeatedly aforesaid operations again, the sediment after cleaning are transferred to spare in 1.5ml EP pipe；
3. the 2. middle drop of final suspension one obtained is placed on glass slide, observe under an optical microscope, if vermiform mite protozoon
Vermiform mite density is 5/cm of > in suspension2, then taken vermiform mite protozoon suspension is qualified；
B. prepare before modeling
1. select the 12 big ear rabbits of week old female regular grades Japan [SCXK(is sweet) 2015~0001] of health, weight 2.0~
Between 2.5kg；
2. preserved skin: rejecting the big ear rabbit back backbone side 4cm × 4cm size area hair of Japan as Experimental Area；
C. modeling
1. selecting injection site, injection site is the test block skin of the big ear rabbit of Japan preserved skin；
2. testing skin area with 75% alcohol disinfecting；
3. by a step, 3. the middle vermiform mite protozoon suspension prepared is transferred in 2ml skin test injection syringe, is drained containing vermiform mite original
Air in the 2ml skin test injection syringe of worm suspension tightens Japanese big ear rabbit local skin, a hand-held syringe needle inclined-plane on the other hand
Upwards, with skin in 5 ° of piercing Japan ear rabbit test block skins greatly, to needle slope completely into intradermal, it is laid flat syringe.With
The fixed pintle of the hand of skin tightening, injects 0.2ml vermiform mite protozoon suspension, makes to be partially formed skin mound, pull out needle rapidly, can not be by
Pressure part.
D. observation method
1. observation and skin lesion scores of erythema
Observation injection site skin lesion changes after injecting vermiform mite protozoon suspension for the first time, and gives 0~4 standard to erythema and comment
Point, it is observed continuously 6 weeks；
2. Histopathological Studies
The big ear rabbit test block dermatosis tissue row pathologic finding of excision Japan, taken tissue is fixed, is embedded, and is carried out normal
Advise HE dyeing and microscopically observation.A skin lesion is cut every 2 weeks, is observed 6 weeks altogether.
Advantages of the present invention:
1. the big ear rabbit of present invention application Japan is cooked animal model, the skin of rabbit is closer with human body, is divided into epidermis, corium and subcutaneously
Tissue, and contain cutaneous appendages, such as: hair follicle and sebaceous glands.Japanese big ear rabbit skin is thicker, is relatively held by intracutaneous injection mode
Easily vermiform mite is injected in skin corium.Japanese big ear rabbit is sensitive to stimulate the reaction, and reaction is similar to people, anti-infective and adaptation rate
Stronger, the death rate is low, and Experimental Area is big, more suitable for the modeling of acne rosacea sample skin lesion caused by vermiform mite.
2. the vermiform mite of the available relative clean of the step of by preparation cleaning vermiform mite protozoon suspension, Bu Huizao
At the big ear rabbit bacterium infection of Japan after intracutaneous injection, the big ear rabbit skin pathological tissue of other impurities interference observation Japan will not influence
Experimental result, reach vermiform mite cause the skin lesion of acne rosacea sample animal model purpose.
3. the present invention by way of intracutaneous injection, can successfully observe Japanese big ear model of rabbit and vermiform mite
The same clinical manifestation of positive acne rosacea patient, furthermore it can be observed that and vermiform mite in Japanese big ear rabbit histopathology
The positive similar granuloma of acne rosacea patient tissue pathology height.Meet Japanese big ear rabbit as vermiform mite and causes rose
The requirement of acneform lesion animal model, and it is able to reflect out the similar clinical and Histopathologic appearance of height.
4. the present invention, according to ethics theory, human experimentation is offensive to morality specification.The present invention can replicate compacted shape on animal
The clinical manifestation of the acne rosacea lesions of patients of the mite positive and Histopathologic appearance can carry out rose caused by vermiform mite for a long time
Continuous pathological study and correlative study on acne animal model.
Detailed description of the invention
Fig. 1 is vermiform mite (the original enlargement ratio in acne rosacea patient facial region's smegma object of the vermiform mite positive
× 40).
Fig. 2A is the visible imflammatory erythema of the big ear rabbit test block back skin lesion of Japan after Human Follicle Mite Infection 1 day.
Fig. 2 B is after Human Follicle Mite Infection 2 weeks about 0.4 on the basis of the big visible erythema of ear rabbit test block back skin lesion of Japan~
0.8cm or so aubergine papule；
Fig. 2 C is that big the back skin lesion of ear rabbit test block is visible is dispersed in 0.4~0.8cm or so inflammatory mound for Japan after Human Follicle Mite Infection 4 weeks
Rash；
Fig. 2 D is that big the back skin lesion of ear rabbit test block is visible is dispersed in 0.3~0.5cm or so tubercle for Japan after Human Follicle Mite Infection 6 weeks.
Fig. 2 E is the vermiform mite positive acne rosacea patients with clinical manifestations that the course of disease is 3 months, be erythema on the basis of about 0.3~
1cm or so papule, tubercle.
Fig. 3 A is the big ear rabbit test block back skin lesion pathological tissue performance of Japan after Human Follicle Mite Infection 4 weeks, is had for intradermal
Inflammatory cell infiltration, granuloma form (original enlargement ratio × 100)；
Fig. 3 B is the big ear rabbit test block back skin lesion pathological tissue performance of Japan after Human Follicle Mite Infection 6 weeks, is had completely for intradermal
Granuloma formed (original enlargement ratio × 100)；
Fig. 3 C is the vermiform mite positive acne rosacea lesions of patients pathological tissue performance that the course of disease is 3 months, is that intradermal inflammatory is thin
Born of the same parents' infiltration and granuloma form (original enlargement ratio × 40).
Fig. 3 D is the partial enlarged view of Fig. 3 C, is that intradermal inflammatory cell infiltration and granuloma form (original amplification
Multiplying power × 100).
Specific embodiment
In the following, being described further again in conjunction with attached drawing to technical solution of the present invention:
A method of building vermiform mite causes acne rosacea sample skin lesion animal model, the steps include:
Construct animal model
1, the selection and culture of the big ear rabbit of Japan
1. selecting the 12 big ear rabbits of week old female regular grades Japan [SCXK(is sweet) 2015~0001] 24 of health, weight exists
Between 2.0~2.5kg.
2. animal packet: after Japanese big ear rabbit is raised 1 week, being randomly divided into experimental group and control group, every group 12, divide cage
It raises in clean animal room.
3. rearing conditions: 23 ± 2 DEG C of temperature, humidity 54 ± 10% divulges information 2~3 times daily, it is small to continue 1 every time
When, according to the natural life pattern of Japanese big ear rabbit, forage nursing is given, so that its is freely drunk water and is fed, move freely,
It raises 1 week with this condition, after waiting Japanese big ear rabbit to adapt to the condition completely, starts experimental implementation.
2, the preparation of experiment cleaning vermiform mite
1. for mite person: the 9th 4 〇 hospital dermatology department out-patient of joint logistics system army, the Chinese People's Liberation Army is diagnosed as rose
Acne, and acarid microscopy is positive.Human Follicle Mite Infection scale standard calculates the quantity of mite referring to skin every square centimeter, is divided into light
3 degree: 1~5 are weighed as low-grade infection (+), 6~10 are grade and moderate infection (++), and 10 the above are severe infection (+++).This hair
The acne rosacea patient of severe Human Follicle Mite Infection is as mite person, (i.e. vermiform mite density is 5/cm of > in bright main selection2),
Fig. 1 is the vermiform mite in acne rosacea patient facial region's smegma object of the vermiform mite positive, and Fig. 2 E is for mite person's face skin lesion
Clinical manifestation, Fig. 3 C are to show for mite person's Histopathological.
2. position of drawing materials: sebum secretion enriches position at nose week, facial skin lesion.
3. vermiform mite suspension liquid and preparation method thereof: for mite person's warm water cleaning skin, wiping dry, use the towel hot compress patient of sterilizing
Facial 2min, so that sufficiently being opened for mite facial pores.Due to vermiform mite under ex vivo situation low temperature resistant non-refractory, therefore it is whole
It is a take mite during, operated in the environment of 20~30 DEG C of room temperature, doctor need to be faced for mite person, it is motionless to face upward about 60 degree angles of head, cures
Teacher takes fingerstall, the acari taking device after holding sterilizing, scrapes bit end crushing failure at high speed patient's nasal fossa, dilator naris or facial skin lesion with acari taking device
Place, extrusion line head sebum creamy white or light yellow (is slightly different) according to for mite different biopsy method color quality, in acquisition
A 2% Amway dish washing liquid of drop is added in sebum secretion object as sebum lytic agent, mixes the glass for being transferred to 5ml to homogeneous latex emulsion shape
In glass test tube, 1 × PBS(HyClone SH30256.01B of 2ml is added) it is diluted, to protect polypide, then blown with ear washing bulb
It beats and mixes, be placed in 4 DEG C of refrigerators and stand natural sedimentation 2h, after abandoning the supernatant in teat glass, 1 × PBS of 2ml is added again
After being diluted and precipitating and abandon supernatant, repeatedly aforesaid operations again.Finally, the sediment after cleaning is transferred to 1.5ml EP
It is spare in pipe；
4. since vermiform mite density can change in operation, therefore the above-mentioned 3. middle drop of final suspension one obtained is placed in load
On slide, in optical microscopy (BX53;Olympus Corporation, Tokyo, Japan) under observe, if vermiform mite is former
Vermiform mite density is 5/cm of > in worm suspension2Then taken vermiform mite protozoon suspension is qualified.
3, modeling method
1. preparing before modeling: marking the big ear rabbit backbone side experiment skin ranges of 4cm × 4cm Japan with marking pen.And it uses
Clean Brush for cleaning dips a small amount of physiological saline, and uniform brush keeps the rabbit hair wet in Japanese big ear rabbit Experimental Area.Preserved skin packet is opened to take
Shaving blade strikes off the Experimental Area rabbit hair out, so that Experimental Area skin sufficiently exposes.
2. modeling:
A. by vermiform mite protozoon suspension, intracutaneous injection is in Japanese big ear rabbit back test block, each skin mound 0.2ml.
B. intracutaneous injection method:
(1) injection site is selected, injection site is the test block skin of the big ear rabbit of Japan preserved skin；(2) with 75% alcohol disinfecting reality
Test skin area；(3) air in the 2ml skin test injection syringe containing vermiform mite protozoon suspension or 1 × PBS is drained, it is taut on the other hand
Tight local skin, a hand-held syringe needle inclined-plane is upward, with skin in 5 ° of big ear rabbit test block skins of piercing Japan, to syringe needle
Inclined-plane is laid flat syringe completely into intradermal.Pintle is fixed with the hand of skin tightening, to the intradermal injection of experimental group test block skin
0.2ml vermiform mite protozoon suspension, injection 0.2ml1 × PBS intradermal to control group test block skin, makes to be partially formed skin mound.It is fast
Speed pulls out needle, not pressable part.
3. observation: experimental group and control group the 1st day and every 2 weeks observation skin lesion variation, and skin histology row disease is cut every 2 weeks
Reason checks.
4. the preparation of skin lesion materials and skin histology paraffin section: materials choose fresh skin lesion tissue, in skin lesion edge
Cut 1.4cm × 1.4cm × 0.2cm size shuttle shape skin lesion row histopathological examination；Taken tissue is molten using 8% formalin
Liquid is fixed, and is successively carried out dehydrating in concentration is 80%, 90%, 95%, 100% alcoholic solution later, saturating using dimethylbenzene
Bright, then paraffin-embedded tissue, with 5~6 μm of thickness serial section, row HE is dyed after dewaxing.
4, observation method
1. observation and skin lesion scores of erythema
Observation each group injection site skin lesion changes after injecting vermiform mite protozoon suspension for the first time, and gives and score to erythema, and 0
It is divided into no erythema, 1 point is erythema of maying be seen indistinctly, and 2 points are clearly visible light red spot, obscurity boundary, and 3 points are erythema is obvious and boundary
Clearly, 4 points are erythema color depth sharpness of border.It is observed continuously 6 weeks.
Only there is inflammation in the big ear rabbit back skin lesion test block of Japan after 1 day in visible experimental group Human Follicle Mite Infection during observation
Disease erythema, injection skin mound region are calmed down, and do not occur the clinical manifestations such as papulonodule (A referring to fig. 2)；The big ear rabbit back of Japan after 2 weeks
Still there is inflammation erythema in portion skin lesion test block, and injection skin mound Regional Red inflamed papules are aubergine (B referring to fig. 2)；It is Japanese after 4 weeks
Big ear rabbit back skin lesion test block inflammation erythema subsides, and injection skin mound region inflamed papules, which do not subside, to be calmed down, and there are pale red inflammation
Property papule, tubercle, touching have activity (C referring to fig. 2)；The big ear rabbit back skin lesion test block of Japan has no inflammation erythema after 6 weeks,
Injection skin mound region papule, tubercle do not subside and calm down, it is seen that papulonodule is color of the leather, inflammatory resolution (D referring to fig. 2).Fig. 2 E institute
It is shown as the vermiform mite positive acne rosacea patients with clinical manifestations that the course of disease is 3 months, is the mound about 0.3~1cm or so on the basis of erythema
Rash, tubercle.Fig. 2 B, Fig. 2 C and Fig. 2 D are that the big ear rabbit of experimental group Japan showed in test block skin lesion in the 2nd week, the 4th week and the 6th week,
In whole process the visible big ear rabbit test block of Japan be dispersed in papule on the basis of imflammatory erythema, tubercle, Fig. 2 B, Fig. 2 C and Fig. 2 D with
Fig. 2 E, which is contrasted, show that Japanese big ear rabbit test block skin lesion clinical manifestation is faced with the occurred skin lesion of vermiform mite positive acne rosacea patient
Bed performance is consistent.
Control group test block skin after intracutaneous injection 1 day did not occurred imflammatory erythema, papule, later the 2nd week, the 4th
Do not occur imflammatory erythema, papule and tubercle skin lesion performance during the entire observation of week and the 6th week.
2. Histopathological Studies
Cut off skin histology row pathologic finding.Taken tissue is fixed, embeds, and carries out conventional H E and dyes and see under the microscope
It examines.A skin lesion is cut every 2 weeks, is observed 6 weeks altogether.
During observing under the microscope, Human Follicle Mite Infection 4 weeks in big ear rabbit damage histopathologic slide, experimental group Japan
Skin lesion intradermal has inflammatory cell infiltration afterwards, and granuloma forms (referring to Fig. 3 A)；Skin lesion intradermal after Human Follicle Mite Infection 6 weeks
There is complete granuloma to form (referring to Fig. 3 B).Fig. 3 C and Fig. 3 D are the vermiform mite positive acne rosacea that the course of disease is 3 months
The performance of lesions of patients pathological tissue, forms for intradermal inflammatory cell infiltration and granuloma.Fig. 3 A, Fig. 3 B are experimental group day
This big ear rabbit visible inflammatory cell infiltration and complete granuloma shape in the 4th week and the 6th week tissue pathological slice
At Fig. 3 C, Fig. 3 D and Fig. 3 A, Fig. 3 B, which are contrasted, obtains Japanese big ear rabbit test block skin lesion pathological tissue performance and vermiform mite positive rose
Rare the occurred Histopathologic appearance of patients with acne is consistent.
Control group does not occur inflammatory during the entire observation in the 2nd week, the 4th week and the 6th week after 1 × PBS of intracutaneous injection
The Histopathologic appearance of cellular infiltration and granuloma.
In conclusion there is the clinical table as the acne rosacea patient of the vermiform mite positive in the big ear model of rabbit of Japan
It is existing, the acne rosacea patient tissue pathology height with the vermiform mite positive is furthermore observed that in Japanese big ear rabbit histopathology
Similar granuloma.Meet the requirement that acne rosacea sample skin lesion animal model is caused using Japanese big ear rabbit as vermiform mite, and
It is able to reflect out the similar clinical and Histopathologic appearance of height.
The present invention further widening roadbed on the Research foundation in the past to acne rosacea and vermiform mite is explored and is excavated, mentions
Viewpoint out: vermiform mite is one of the main pathogenic of acne rosacea.Due to cannot achieve continuous biopsy action in tissue
State Histopathological Studies limit further research of the vermiform mite to acne rosacea.Therefore, one aspect of the present invention constructs vermiform mite
The big ear model of rabbit of the Japan of caused acne rosacea sample skin lesion, come further study vermiform mite cause acne rosacea the cause of disease and
Pathogenesis, while a kind of new way can be provided for the diagnoses and treatment of acne rosacea.On the other hand it can be used for the following related rose
The evaluation studies of rare acne drug.
Claims (1)
1. a kind of method that building vermiform mite causes acne rosacea sample skin lesion animal model, the steps include:
A. the preparation of vermiform mite protozoon suspension is cleaned
1. for mite person: the 9th 4 〇 hospital dermatology department out-patient of joint logistics system army, the Chinese People's Liberation Army is diagnosed as rose
Acne, and acarid microscopy is positive；
2. preparation method: for mite person's warm water cleaning skin, wiping dry, then squeeze the scraping wing of nose or facial sebum point using acari taking device
2% dish washing liquid of drop is added in the sebum secretion object of acquisition, mixes to homogeneous latex emulsion shape, and homogeneous latex emulsion shape is shifted for secretion
Into the teat glass of 5ml, 1 × PBS that 2ml is added is diluted, and is blown and beaten and is mixed with ear washing bulb, is placed in 4 DEG C of refrigerators and is stood certainly
So precipitating 2h, after abandoning the supernatant in teat glass, 1 × PBS that 2ml is added again is diluted and precipitates, and abandons supernatant
Afterwards, repeatedly aforesaid operations again, the sediment after cleaning are transferred to spare in 1.5ml EP pipe；
3. the 2. middle drop of final suspension one obtained is placed on glass slide, observe under an optical microscope, if vermiform mite protozoon
Vermiform mite density is 5/cm of > in suspension2, then taken vermiform mite protozoon suspension is qualified；
B. prepare before modeling
1. selecting the 12 big ear rabbits of week old female regular grade Japan of health, weight is between 2.0~2.5kg；
2. preserved skin: rejecting the big ear rabbit back backbone side 4cm × 4cm size area hair of Japan as Experimental Area；
C. modeling
1. selecting injection site, injection site is the test block skin of the big ear rabbit of Japan preserved skin；
2. testing skin area with 75% alcohol disinfecting；
3. by a step, 3. the middle vermiform mite protozoon suspension prepared is transferred in 2ml skin test injection syringe, is drained containing vermiform mite original
Air in the 2ml skin test injection syringe of worm suspension tightens Japanese big ear rabbit local skin, a hand-held syringe needle inclined-plane on the other hand
Upwards, with skin in 5 ° of piercing Japan ear rabbit test block skins greatly, to needle slope completely into intradermal, it is laid flat syringe；
Pintle is fixed with the hand of skin tightening, 0.2ml vermiform mite protozoon suspension is injected, makes to be partially formed skin mound, pull out needle rapidly,
Not pressable part；
D. observation method
1. observation and skin lesion scores of erythema
Observation injection site skin lesion changes after injecting vermiform mite protozoon suspension for the first time, and gives 0~4 standard to erythema and comment
Point, it is observed continuously 6 weeks；
2. Histopathological Studies
The big ear rabbit test block dermatosis tissue row pathologic finding of excision Japan, taken tissue is fixed, is embedded, and is carried out normal
Advise HE dyeing and microscopically observation；
A skin lesion is cut every 2 weeks, is observed 6 weeks altogether.
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| CN111534490A (en) * | 2020-05-11 | 2020-08-14 | 中南大学湘雅医院 | Model construction method for treating acne rosacea by hydroxychloroquine |
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| CN110624102A (en) * | 2019-11-08 | 2019-12-31 | 罗洋 | Clinical application of bevacizine as medicine for repairing demodex disease skin damage |
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