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Ukusekwa kweemodeli zezilwanyana zotshintsho lwemodic (MC) sisiseko esibalulekileyo sokufunda i-MC. Amashumi amahlanu anesine i-zealand imivundla emhlophe yahlulahlulwa liqela lokusebenza le-sham, iqela lemisipha (iqela le-MPUTE) kunye neQela lokuKhupha i-nucleuse (i-NPEUS). Kwiqela le-NPO, i-InterverEibral ityhilwe yi-anterolated indlela ye-anterolated lumbar ye-Lumbar kwaye inaliti yayisetyenziselwa ukugoba umzimba we-l5 vertebral kufutshane neplate yokuphela. I-NP ikhutshwe kwi-L1 / 2 i-interverpeibral i-interpile ngesirinji kwaye igcinwe kuyo. Ukuqhuba umngxunya kwithambo le-subchollral. Iinkqubo zoqhaqho kunye neendlela zokuqhuba kwiqela lemisipha yeqela lemisipha kunye neqela lokuSebenza leSham lalifana nezo kwiqela le-NP. Kwiqela le-M, isiqwenga somsipha sabekwa emngxunyeni, ngelixa kwiqela le-Sham-Proper-sham, akukho nto yayibekwe emngxunyeni. Emva kotyando, ukuskena i-MRI kunye nokuvavanywa kwemolekyuli yemolekyuli kwenziwa. Umqondiso kwiqela le-NPE watshintsha, kodwa kwakungekho tshintsho lukhazimla kwiqela leSham kunye neqela le-MARD. Ukuqatshelwa kwembali kubonakalise ukuba ukwenziwa kwe-tisferity engaqhelekanga kwaqwalaselwa kwindawo yokubekwa indawo, kwaye intetho ye-Il-4, il-17 ne-In-γ inyuke kwiqela le-NPO. Ukufakwa kwe-NP kwithambo le-subchotral kunokwenza imodeli yezilwanyana ye-MC.
Utshintsho lweModeli (MC) zizilothi ze-vertebral zokuphela kwe-vertebral kunye nethambo elikufuphi ne-bone elibonakalayo kwi-magnetic resing (MRI). Zixhaphakile kakhulu kubantu abaneempawu ezinxulumene noko. Izifundo ezininzi zigxininise ukubaluleka kwe-MC ngenxa yomanyano lwayo ngentlungu esezantsi (LBP) 2,3. I-De Roos et al.4 kunye ne-MODIC ET Al.5 Ngeendlela zaqala zachaza iintlobo ezintathu ezahlukeneyo zophawu lwe-subcholral kwi-vertebral yethambo. Uhlobo lomfo oguqukayo luyi-hypoinse kwi-T1-ye-T1W) Esi silon sityhila iziphelo ze-fissure kunye ne-vascing ye-vascular ye-vascular ebukhali kwithambo lomongo. Uhlobo lomfo oguqukayo ubonisa uphawu oluphezulu kwi-T1W kunye nolandelelwano lwe-T2W. Kolu hlobo lwesilonda, intshabalalo yesiphelo inokufunyanwa, kunye nokufakelwa kwembali ye-thewological yomongo obufuphi. Uhlobo lomfo kwi-III III ibonisa umqondiso ophantsi kwi-T1W kunye ne-T2W ulandelelwano. Izilonda zekratshi ezihambelana neziphetho ziqwalaselwe6. I-MC ithathelwa ingqalelo isifo sendalo somqolo kwaye sihambelana nezifo ezininzi ze-sprine7,8,9.
Ukuqwalaselwa kwedatha ekhoyo, izifundo ezininzi zibonelele ngeenkcukacha ezigcweleyo kwi-Etioloss kunye neendlela zendalo zeMC. UAlbert Et Al. Icebise ukuba i-MC inokubangelwa kukucutha i-dic8. Hu Al. I-MC ye-MC kwi-diegeneration enkulu ye-Dead. I-KROC icetywe umxholo we "disc yangaphakathi yokuqhekeka," echaza ukuba i-tem ye-disc ephindaphindwayo inokukhokelela kwimicrotere kwi-Ektrote. Emva kokusekwa kwe-cleft, ukupheliswa okupheliselwe yi-pundeus pulposus (NP) inokubangela impendulo ye-autommune, ekhokelela kuphuhliso lwe-MC11. Mat Al. Babelane ngombono ofanayo kwaye waxela ukuba i-aut-i-autoimmunity idlala indima ephambili kwi-pathogenesis ye-MC12.
Iiseli zenkqubo yomzimba, ngakumbi i-CD4 + T I-LILMPHICY, idlale indima ebalulekileyo kwi-pathogenesis ye-autoimmunity13. Iseti ye-th17 isandula ukufunyanwa i-th17 i-th17 iseti yeseti ye-proinfine il il il il il il il il il il il il il il il il il il il il il il il ye-Il, 17, ikhuthaza i-chemokine ibyct, kwaye ivuselele iiseli ze-TMOTIIN, kwaye zivuselele iiseli ze-T Iiseli ze-th2 zidlala indima ekhethekileyo kwi-pathogenesis yempendulo yomzimba. Inkcazo ye-Il-4 njengeseli kwi-th2 ye-th2 inokuthi ikhokelele kwiziphumo ezinzima ze-ihumopathological15.
Nangona izifundo ezininzi zeklinikhi zenziwe kwi-MC16,17,18,19,20,21,21,22,22,22,24, kusekho ukunqongophala kweemodeli ezifanelekileyo ze-MC ezinokuthi zibekho rhoqo ebantwini kwaye zinokuba njalo isetyenziselwa ukuphanda i-Entimotoy okanye unyango olutsha olunje ngonyango olujolise kuyo. Ukuza kuthi ga ngoku, zimbalwa kuphela iimodeli zezilwanyana ze-MC zixeliwe ukuba zifunde indlela ye-pathoomagicals yendalo.
Ngokusekwe kwi-Autoimmune ithiyori ecetywayo ngu-Albert ne-Ma, lo phononongo useke imodeli elula neyokuzala ye-MC nge-Autotransplant Np kufutshane neplate yokuphela kwe-vertebral. Ezinye iinjongo kufuneka ziqwalasele iimpawu zemodeli yezilwanyana kwaye zihlole iindlela ezithile ze-NP ekuphuhlisweni kwe-MC. Ukuza kuthi ga ngoku, sisebenzisa iindlela ezifana ne-molecular biology, i-MRI, nakwizifundo ze-chastolos zenza ukuqhubela phambili kwe-MC.
Imivundla emibini yabulawa ngabopha ngexesha lotyando, kwaye imikhweli emine yafa ngexesha le-anesthesia ngexesha leMRI. Imivundla eseleyo 48 yasinda kwaye ayibonisanga imiqondiso yokuziphatha okanye imiqondiso emva kotyando.
I-MRI ibonisa ukuba ubukhulu bemiqondiso yezicubu ezifakelweyo kwimingxunya eyahlukileyo. Ubuninzi besiginali kwiqela le-L5 vertebral kwiqela le-NTE ngokuthe ngcembe batshintsha nge-12, 16 neeveki ezingama-16 ukuya kwe-20 emva kokufakelwa komqondiso (i-T1W babonisa uphawu oluxubileyo), ngelixa i-MRI Emanye amaqela amabini eenxalenye ezifakelweyo zihlala zizinzile ngexesha elinye (Fig. 1A, b).
. Akukho zimpawu zingaqhelekanga zifumaneka kwimifanekiso yeqela leNtsebenziswano yeSham. . . Ukususela kwiveki yeveki ye-12 ukuya kwiveki yeveki yama-20, imiqondiso ephezulu ye-sporadic ejikeleze imiqondiso esezantsi kwi-T2W ukulandelelana.
I-hynerpia yethambo elibonakalayo inokubonwa kwindawo yokubekwa kwe-vertebral kwiqela le-vertebral kwiqela le-NTE, kwaye i-Hone Hypelasia yenzeka ngokukhawuleza ukusuka kwi-12 ukuya kwi-20c ibonwa kwi-vertebral emodareyitha imizimba; Iqela le-sham kunye nam liqela (Fig. 2c) 2a, b).
. (B) Ubume besiza esifakwe kum iqela (C) I-Bone HyperPisia yenzeka kwindawo efakwe kwiqela le-NPE. I-HyperPlasia yethambo yanda ngokukhawuleza kwaye yandiswa kwi-interverteibral disc kumzimba we-vertebral verteral.
Uhlalutyo lwembali lubonelela ngolwazi oluthe kratya malunga nokuqulunqwa kwethambo. Umzobo 3 ubonakalisa iifoto zamacandelo e-Postoperaperapeative ezenziwe nge-H & E. Kwiqela le-Sham-sham, i-chondrocytes yayilungiselelwe kakuhle kwaye akukho ntshukumo yeseli ifunyenwe (Fig. 3a). Imeko ethile kum yayifana nelo kwiqela leNtsebenziswano (Fig. 3b). Nangona kunjalo, kwiqela le-NPO, inani elikhulu lokhetho kunye nokwanda kweeseli ze-NP-ezinjengee-NP ezigcinwe kwindawo yokubekwa kwempahla (i-3C.
. (B) imeko yendawo yokufakelwa kwiqela kum iyafana neqela le-sham. I-Trabecusulae kunye ne-chondrocytes zinokubonwa, kodwa akukho ncedo lucacileyo kwindawo yokubekwa kwempahla (ama-40). .
Imbonakalo ye-UNECLEKIN 4 (il-4) Mnuna, i-Enterleukin 17 (il-17) Mrna, kunye ne-Interferon γ (i-Itn-γ) I-MPI kunye nam. Xa amanqanaba enzalo lwezinto ezithagethiweyo athelekiswa, i-gene bibs, il-17, kunye ne-IFN-17, yanda kakhulu kwiqela le-NTO xa kuthelekiswa neqela le-NTO kunye neqela le-Sham. 4) (P <0.05). Xa kuthelekiswa neQela leNtsebenzo yeSham, amanqanaba e-IL-4, il-17, kunye ne-IFN-17 kwiqela le-MADY inyuke kancinci kwaye engafikanga tshintsho lweenkcukacha-manani (P> 0.05).
Imbonakalo ye-MRNA ye-Il-4, il-17 ne-Ill-γ kwiqela le-NPO ibonisa imeko ephezulu kakhulu kunaleyo ikwiqela le-Sham kunye neqela le-sham (p <0.05).
Ngokwahlukileyo, amanqanaba entetho ebinzana nam abonakalise umahluko obalulekileyo (P> 0.05).
Uhlalutyo lwe-Blot Western lwenziwa kusetyenziswa i-antibodies ezifumanekayo kwezorhwebo ngokuchasene ne-Il-4 kunye ne-Il-17 ukuqinisekisa ipatheni eguqulweyo. Njengoko kubonisiwe kwi-GAGES 5A, B, xa kuthelekiswa neqela le-MAM kunye neQela leNtsebenziswano, iProtein yeProtein ye-IL-4 kunye ne-Il-17 kwiqela le-NTE lalinyuswe kakhulu (p <0.05). Xa kuthelekiswa neQela leNtsebenzo yeSham, amanqanaba eproteni ye-IL-4 kunye ne-Il-17 kwiqela le-Me-17 kwiqela eliphumelelayo lokufikelela kwiinguqu ezibalulekileyo (P> 0.05).
. (B) I-virtogram ye-West West.
Ngenxa yenani eliqingqiweyo lesampulu yomntu efunyenwe ngexesha lotyando, ecacileyo neneenkcukacha kwi-pathogenesis ye-MC inzima. Sizame ukuseka imodeli yezilwanyana ye-MC yokufunda indlela yayo ye-Phyphogical. Kwangelo xesha, uvavanyo lwe-radiralogical, uvavanyo lwembali kunye novavanyo lwe-molecular biologional lwasetyenziswa ukulandela ikhosi ye-MC ebangelwa yi-autograft ye-NP Autgraft. Ngenxa yoko, imodeli yokufakelwa kwe-NP yakhokelela kutshintsho ngokuthe ngcembe kumqondiso ukusuka kwi-12-veki ukuya kwiveki yamaxesha angama-20 (i-Tract exubeneyo kwi-T1W ngokulandelelana), ebonisa i-tismologings kunye nemolekyuli Uvavanyo lwebhayoloji luqinisekisile iziphumo zesifundo se-radiological.
Iziphumo zale milingo zibonisa ukuba utshintsho olubonakalayo nolwenziweyo lwenzeka kwindawo yolwaphulo-mthetho lwe-vertebral kwiqela le-NTE. Kwangelo xesha, intetho ye-Il-4, il-17 ne-Il-4, il-17, i-Il-17 neBon-1 kwaye iBon-1 ye-Autcle i-chalteus Umzimba unokubangela uthotho lotshintsho lomqondiso kunye ne-morphological. Kulula ukufumana ukuba iimpawu zomqondiso zemozulu ye-vertebral yemodeli ye-T1W, i-TALL NE-SINTAL kunye ne-TOPIL ePhakathisiweyo kwi-T2W ukulandelelana kwe-T2W) zifana ne-MRI Qinisekisa ukuqaphela kwe-sumelogy kunye ne-anatomy ebonakalayo, oko kukuthi, utshintsho kwiiseli zomzimba ze-vertebral ziyaqhubeka. Nangona impendulo yokuvuvukala ebangelwe kukungabonakali kabuhlungu emva kokuba itshitshiswe, iziphumo ze-MRI zibonisa ukuba utshintsho olwenziweyo luye lwaveza iiveki ezili-12 emva kokubuyela kwi-20 ngaphandle kweempawu zokubuyisela kwimeko yesiqhelo. Ezi ziphumo zibonisa ukuba i-Autortebraus vertebral i-NP yindlela ethembekileyo yokuseka i-MV ye-MV kwimivundla.
Le modeli yokuhlwayelwa ifuna ubuchule obufanelekileyo, ixesha, kunye nokuzama utyando. Kwimvavanyo yangaphambili, ukuguqula okanye ukukhuthaza kakhulu kwezakhiwo ze-parversebratus zinokukhokelela ekumiselweni kwe-vertebrall osteophytes. Inkathalo kufuneka ithathwe ukuba ingonakalisi okanye icaphukise iidiski ezikufuphi. Kuba ubunzulu bokungena kufuneka bulawulwe ukufumana iziphumo ezingaguqukiyo kunye nezibuyisiweyo, sizenzile iplagi ngokusika i-sheath ye-3 mm inaliti ye-3 mm i-3 mm inaliti i-3 mm inaliti. Ukusebenzisa le plug kuqinisekisa ubunzulu bokuthabatha iyunifomu kumzimba we-vertebral. Kuvavanyo lwangaphambili, oogqirha abathathu bamathambo abandakanyeka ekusebenzeni bafumana iinaliti eziyi-16 kube lula ukuba basebenze nge-18-gegenes inaliti okanye ezinye iindlela. Ukuthintela ukopha ngokugqithisileyo ngexesha lokuqhuba, ukubamba inaliti okwethutyana kuya kubonelela ngomngxunya ofanelekileyo wokufakelwa, kucebisa ukuba inqanaba elithile le-MC lingalawulwa ngale ndlela.
Nangona izifundo ezininzi zijolise i-MC, kancinane ziyaziwa malunga ne-Etioloss kunye ne-pathogenesis ye-MC25,26,27. Ngokusekwe kwizifundo zethu zangaphambili, safumanisa ukuba ukuhamba ngokuzenzekelayo kudlala indima ephambili kwindawo yokwenzeka kunye nophuhliso lwe-MC12. Olu phando luvavanye inkcazo yobungakanani be-IL-4, il-17, kunye ne-IFN-17, eyiyona ndlela iphambili ye-CD4 + iiseli ze-CD4 + emva kokuvuselela kuka-Antign. Kwisifundo sethu, xa kuthelekiswa neqela elingelolize, iqela le-Npe lalibonisa i-Il-4, i-Il-17, kunye ne-IFN-17, kunye namanqanaba eproteni ye-IL-4 kunye ne-Il-17.
Iklinikhi, i-Il-17 i-Mr-17 i-Mrna inyuswe kwiiseli ze-NP kwizigulana ze-disc28. Ukunyuka kwamanqanaba e-Il-4 kunye ne-IFN-γ afumaneka kwimodeli ye-CDI ye-CO-AM-AYIRRORE ye-CORCRORE ithelekiswa nolawulo lwempilo29. I-Il-17 idlala indima ephambili kwi-insulamition, ukwenzakala kwezicubu kwizifo ze-autoimMMMMMMMMMMMUNO33 kwaye iphucule impendulo yomzimba kwi-On-¶31. I-Il-17 i-Il-17-quiste ye-Il-17 inikwe i-Fissies yeTrissi / LPR ML / LPR I-MECE32 kunye ne-Autoimmununity-SureChoptity i-mice33. I-Il-4 inokuthi ithintele intetho ye-proinfines cytokines (njenge-Il-1 β kunye ne-tnfcy) kunye ne-macrophage34. Kwaxelwa ukuba intetho ye-MRN-4 yahlukile kwiqela le-NPO xa kuthelekiswa ne-IL-17 kunye ne-IFN-γ ngexesha elinye; Imbonakalo ye-MRNA ye-IFN-γ kwiqela leNTE laliphezulu kakhulu kunolo nakwelinye amaqela. Ke ngoko, ukuba i-IT-γ Imveliso inokuba ngumlamli wempendulo ye-inflommatory ebanjiwe yi-NP Contey. Izifundo zibonisile ukuba i-IFN-γ iveliswa ziintlobo zeseli ezininzi, kubandakanya uhlobo olwenziweyo lwe-1 iiseli ze-T, iiseli zokupheka ezenziwe ngendalo, iiseli zomncedisi wendalo, kunye ne-macrophas35,36, kwaye sisitshixo seCytokine ephambili ekhuthaza iimpendulo zomnumli.
Olu phononongo lubonisa ukuba impendulo ye-autommune inokubandakanyeka kwisehlo kunye nophuhliso lwe-MC. Luoa et al. ifumanise ukuba iimpawu zomqondiso ze-MC kwaye ivelise i-NP ifana kwi-MRI, kwaye zombini zibonisa uphawu oluphezulu kwi-T2W ukulandelelana 38. Ezinye ii-cytokines ziqinisekisiwe ukuba zinxulunyaniswa ngokusondeleyo ne-MC, ezinjenge-IL-139. Mat Al. Icebise ukuba ukupheliswa kwe-NP okanye ezantsi kwe-NP kunokuba nefuthe elikhulu kwisehlo kunye nophuhliso lwe-MC12. UBobechko40 noHerzen et al.41 baxele ukuba i-NP yi-tishu engafakwanga engenakungena ekusasazeni i-vascular ngokuzalwa kwangokuzalwa. I-NP ukuphuphuma kwazisa imizimba yangaphandle eluntwini, ngaloo ndlela benza impilo yokuphendula kwe-Autommune yasekhaya42. Ukuphendula ngokuzenzekelayo kunokuphembelela inani elikhulu lezinto zomzimba, kwaye xa le nto iqhubeka ngokuqhubekayo kwizicubu, zinokubangela utshintsho ku-Local Low43. Kolu phononongo, i-orexpression ye-Il-4, il-17 kunye ne-IFN-γ yimiba yomzimba oqhelekileyo, iqinisekisa ubudlelwane obusondeleyo phakathi kwe-NP kunye ne-MCS44. Le modeli yesilwanyana inyibilike kakuhle i-NP ye-NP ye-NP kunye nokungena kwiplate yokuphela. Le nkqubo ityhila ifuthe lokutsala i-autoimmunity kwi-MC.
Njengoko bekulindelwe, le modeli yesilwanyana isinika iqonga elinokwenzeka lokufunda i-MC. Nangona kunjalo, le modeli isenemida emiselweyo: Okokuqala, ngexesha lesigaba sokuqwalaselwa kwesilwanyana, iSigaba sokuPhayithisa esiPhakathi kufuneka sikhutshwe ngovavanyo lwendalo kunye nemolekyuli, ke ezinye izilwanyana "ziwela ixesha. Okwesibini, nangona amanqaku amathathu asekwe kolu phononongo, ngelishwa, sisondela kuphela uhlobo lwe-MC (uhlobo lwe-modic kuphela), ngoko ke akwanele ukumela inkqubo yokuphuhliswa kwesifo, kwaye amanqaku amaninzi kufuneka asetelwe Nceda qaphela yonke utshintsho lomqondiso. Okwesithathu, utshintsho kubume bezicubu lunokuboniswa ngokucacileyo sisitayile sembali, kodwa ezinye iindlela ezikhethekileyo zinokuchaza ngcono utshintsho oluncinci kule modeli. Umzekelo, i-microscopy ye-microscopy ye-microscopy yasetyenziselwa ukuhlalutya ukwenziwa kwe-fibrocarilage kwi-intervertibral crialterals45. Iziphumo zexesha elide ze-NP kwi-MC kunye nesiphelo sifuna ukuqhubeka nophando.
Amashumi amahlanu anesine yendoda entsha ye-zealand emhlophe (ubunzima malunga ne-2,5-3 kg, iminyaka emi-3-3.5 yenziwa ngokulandelelana kwiqela le-sham ye-sham, iqela lemisipha) kunye neqela le-nerve Zonke iinkqubo zokuvavanya zamkelwa yikomiti yokuziphatha ye-Tianjin, kwaye iindlela zokuvavanywa zenziwa ngokuhambelana nezikhokelo ezivunyiweyo.
Olunye uphuculo lwenziwe ngeendlela ze-S. Sobajima 46. Umvundla ngamnye wabekwa kwindawo yokuphinda-phinda i-Landal kunye ne-ANTARE YEMPENDULO EZIQHELEKILEYO ZE-Consteibral Cons (iVDS) yavezwa kusetyenziswa i-Postroperitol Adpoitoneal cindo. Umvundla ngamnye wanikwa i-Anesthesia jikelele (i-20% urethane, i-5 ml / kg ngendlebe). Isikhumba sesikhumba sexesha elide senziwe ukusuka kumda ongezantsi weembambo ukuya kwi-pelvic brim, 2 cm ventral ukuya kwimisipha ye-paraversebral. I-spine yasekunene ye-anterolateral ukusuka kwi-L1 ukuya kwi-L6 ivezwe yi-Sharp kwaye ishukumise i-tissunt ye-tishu engaphezulu, i-retroperitoneal tishu, kunye nemisipha ye-7A). Inqanaba le-disc lagqitywa kusetyenziswa i-pelvic brim njengophawu lomhlaba we-L5-L6. Sebenzisa inaliti ye-16-ye-gauge phumela inaliti yokuqhuba umngxunya kufutshane neplate yokuphela kwe-L5 Vertebra ukuya kwi-3 mm (Fig. 6B). Sebenzisa i-5-ml i-5-ml i-sringee yokukhuthaza i-autcleus nuprousis kwi-L1-L2 i-Interver-L2 i-IntercEibral i-Interct (Fig. 6C). Susa i-publeus pulposus okanye umsipha ngokweemfuno zeqela ngalinye. Emva kokuba i-Drill Hole ikhuliswe kakhulu, i-study efihliweyo ibekwe kwi-fascia enzulu, i-superficialial fascia kunye nolusu, ukhathalela ukungayonakalisi izicubu ze-vertebral ngexesha lotyando.
. . (C) i-autologous mid iyavunwa.
I-Anesthesia Jikelele ilawulwa nge-20% irethane (i-5 ml / kg / kg) ilawulwa nge-vein yendlebe, kunye nee-radinegraphys zentsimbi zaphindwa nge-12, 16, kunye neeveki ezingama-20.
Imivundla inikelwe yinaliti ye-intramuscular ye-ketamine (25.0 mg / kg) kunye ne-Intravenous ye-Intobarbium petobarbital (1.2 g / kg) ngo-12, 16 kunye neeveki ezingama-16 ukuya emva kotyando. Yonke i-spine isuswe kuhlalutyo lwendalo kunye nohlalutyo lokwenyani lwenziwa. Ukuthumela okungalunganga (i-RT-qpcr) kunye nokuncitshiswa kwentshona kwasetyenziselwa ukufumana utshintsho kwimizimba yomzimba.
Iimviwo ze-MRI zenziwa kwimivundla usebenzisa i-3.0 t zeklinikhi yeklinikhi (i-Ge yezonyango, iFlorence, i-SC) ixhotywe nge-orthogonal yelungu. Imivundla yayiyenzelwe i-20% irethane (i-5 ml / kg) nge-vein yendlebe emva koko yabeka isiqhulo ngaphakathi kwe-magnet kunye ne-Inch ye-Inch Deameter Coiyithi ye-5 ye-Inch Deameter (ye-GE yezonyango). I-TREAL B2 I-TRE, 1445 Nk. I-Sagittal I-T2-I-S2-I-S2-I-S2-I-S2-I-TRECINGY I-TREALS ifunyenwe kwezi zicwangciso zilandelayo: I-TREAT SPINIT kunye nexesha eliphindaphindwayo (i-TR) ezingama-2200 i-70M, i-ECE) (TE) Ibala le-Fible lama-260 kunye neSibhozo Sukeri; Ubungqingqwa bokusika yayiyi-2 mm, umsantsa yayingu-0,2 mm.
Emva kwefoto yokugqibela yathatyathwa kwaye umvundla wokugqibela wabulawa, i-sham, i-sham, imisipha emiselweyo, kunye nee-CDS ze-NP zasuswa kuviwo lwengqondo. Izicubu zazilungisiwe kwi-10% ye-Inhloral Afffrated Fitfin kwiveki e-1, ichaziwe kwi-Ethyledine-Mediaminetetratecthotit acid, kunye neparafini yeparafini. Iibhloko zezicubu zifakwe kwiparafini kwaye zisikwe kwicandelo le-sagittal (5 μm) isebenzisa i-rototome. Amacandelo ayenziwa nge-hematoxylin kunye ne-Eosin (H & E).
Emva kokuqokelela iidigre ze-Intervertebral kwiqela ngalinye, i-RNA iyonke yakhutshwa kusetyenziswa ikholamu ye-Uniq-10 (Shanghai Sangnoy ye-Biotechnology Co., i-shanghai) ngokwemiyalelo yomenzi kunye ne-IORMEGA II I-II Refter System (PROMGA) , Madison, Wi, i-USA). Ukubuyela umva kwenziwa.
I-RT-QPCR yenziwa kusetyenziswa i-prism 7300 (esetyenzisiweyo I-Inc. Umthamo we-PCR yokuphendula yayiyi-20 μl kwaye iqulethwe i-1.5 μm ye-CDNA ebisiweyo kunye ne-0,2 μm ye-primer nganye. I-Primers yenziwa nguMligostlest Loyile (i-Reviitrogen, iValencia, ca) kwaye yenziwe yi-Nanjing Stewart biodexnology Co. (China) (i-LTD Ezi meko zihamba ngebhayisikile ye-thermal ye-thermal ye-thermmeral ye-thermmerase kwi-94 ° C kwi-2 min, i-40 yemizuzu nganye kwi-94 ° C yokuqulunqwa kwe-template, i-1 ° C, i-fluorescence. Imilinganiselo yenziwa i-1 min ngo-72 ° C. Zonke iisampulu zaphakanyiswa kathathu kwaye ixabiso eliphakathi lasetyenziselwa uhlalutyo lwe-RT-qpcr. Idatha ye-Amilffied yahlalutywa kusetyenziswa i-flexstation 3 (izixhobo ze-molecular, i-sunnyvale, ca, e-USA). I-Il-4, il-17, kunye ne-Ifn-○ intetho eqhelekileyo yagqitywa kulawulo lwe-Endogenaus (I-ACHB). Amanqanaba abonisa inqanaba lethagethi I-MRNA ibalwa kusetyenziswa indlela ye-2-δδ.
Iprotheyini iyonke yakhutshwa kwizicubu ezisebenzisa i-tissue ye-tipp ye-ripa luffer (equlathe i-proase kunye ne-phosprigue) i-4,000 ye-phospm i-4,000 nge-4 ° C ukususa inkunkuma ye-tissure. I-micrograms engamashumi amahlanu eProtein yayilayishwe ngakwindlela nganye, eyahlulwe yi-10% yephepha le-SDS-teds, emva koko idluliselwe kwi-PVDF membrane. Ukuthintela kwenziwa kwiipesenti ezi-5 zobisi olwenziweyo kwi-Tris-Bulnered Saline (TBS) equkethe i-0.1% phakathi kwe-20 ye-1 h kubushushu begumbi. I-membrane ifakwe kwi-Anti-Decorin ePrayimari (i-1: 200; i-biling, i-china) ngobusuku nge-4 ° China) nge-4 ° China) ngeentsuku yesibini; Ngomntu onesibini olwesibini (ibhokhwe anti-umvundla u-Innoglobulin g kwi-1: 40,000) idityaniswe nehashe ihashe (i-boster, i-china) kwiyure yegumbi. Iimpawu ze-wlot zasentshona zifunyenwe ngokunyusa i-shemilumence kwi-chemilumpceces membrane emva kwe-it-ray iqula. Ngohlalutyo lwe-denstitametric, iibhloko zaziskenwe kwaye zisetyenziswe kusetyenziswa isoftware ye-Bands-iziphumo kwaye iziphumo zavakaliswa njengomyinge wokujongwa kwethagethi kwi-tubingelin i-tunorective.
Iinkcukacha-mabala ezibalwayo zenziwa kusetyenziswa i-SPSSS16.0 Iphakheji yesoftware (SPSS, e-USA). Idatha eqokelelweyo ngexesha lophononongo ibonakalisiwe njengoko kuthetha ukuphambuka komgangatho (kuthetha ± SD) kwaye kuhlalutywe kusetyenziswa iindlela zohlalutyo (i-anova) ukumisela umahluko phakathi kwala maqela mabini. I-P <0.05 ithathelwa ingqalelo njengezibalo.
Ke, ukusekwa kwemodeli yesilwanyana ye-MC ngokufaka i-AutLogous Nps kwi-vertebral yomzimba nokwenza uhlalqweni lwe-macroanatomical, uhlalutyo lwe-MRI lube sisixhobo esibalulekileyo sokuvavanya nokuqonda iindlela ze-MC zabantu kunye nokuphuhlisa unyango Ungenelelo.
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IXESHA LOKUQALA: I-DEC-13 ukuya ku-2424