Kev kho pob zeb hauv raum: G. Fruticosus
Mar 22, 2022
Hu rau: Audrey Hu Whatsapp / hp: 0086 13880143964 Email:audrey.hu@wecistanche.com
PART Ⅰ: Kev ntsuam xyuas ntawm kev ua tau zoo ntawm G. fruticosus hnyav rho tawm hauv kev sim ntxias nephrolithiatic Wistar txiv neej nas
Tilahun Alelign, Tesfaye Sisay Tessema, Asfaw Debella thiab Beyene Petros
Keeb kwm
Urolithiasis(pob zebhauv cov zis) cuam tshuam txog 12 feem pua ntawm cov neeg hauv ntiaj teb nyob rau qee theem hauv lawv lub neej [1]. Lwm cov kev tshawb fawb qhia tau hais tias lub ntiaj teb no prevalence ntawmlub raum pob zebnyob nruab nrab ntawm 2 mus rau 20 feem pua [2, 3]. Urolithiasis cuam tshuam rau txhua lub hnub nyoog, poj niam txiv neej, thiab haiv neeg [4,5], tab sis nws tshwm sim ntau zaus hauv cov txiv neej dua li cov poj niam hnub nyoog 20 txog 49 xyoo [6]. Lub pathogenesis ntawmlub raum pob zebtseem nkag siab tsis tiav [7] thiab nws txoj kev kho mob tseem yog qhov nyuaj, txawm hais tias muaj cov kev kho mob niaj hnub suav nrog ESWL[8]. Cov xwm txheej uas tshwm simpob zebKev tsim muaj xws li nucleation, kev loj hlob, kev sib sau ua ke, thiab khaws cia ntawm cov muaju hauv ob lub raum [9, 10].
Lub raum pob zebtsim yuav tsum tsis tu ncua khaws cia ntawm crystals nyob rau hauv lublub raumtom qab ua tiav cov txheej txheem crystallization[11, 12]. Kev sib cuam tshuam ntawm COM crystals nrog rau saum npoo ntawm lub raum epithelial hlwb yog ib qho tseem ceeb pib tshwm sim hauv nephrolithiasis [13]. Crystal-cell inter-actions ua rau lub zog ntawm cov muaju los ntawm sab basolateral ntawm cov hlwb mus rau hauv qab daus daim nyias nyias [14]. Ib txoj kev tshawb fawb ntawm cov qauv tsiaj kuj qhia tau hais tias kev tswj hwm ntawm CaOx crystals lossis oxalate ions zoo li muaj tshuaj lom, ua rau lub raum tubular cell puas [7]. Kev raug rau qib siab dua ntawm oxalate lossis CaOx crystals induces epithelial cell raug mob, uas yog ib qho tseem ceeb rau cov tom ntej.pob zebkev [15, 16].
Ib qho tsis sib npaug ntawmtso zispob zebinhibitors thiab txhawb nqa tau pom tias yog qhov ua raupob zebkev tsim kho [17]. Cov txhawb pab txhawbpob zebtsim [18], tab sis inhibitors txo qhov pib ntawm supersaturation, nucleation, crystal kev loj hlob, thiab tus nqi ntawm aggregation [17]. Txawm li cas los xij, inhibitors ua haujlwm sib npaug rau txhua tus; yog li ntawd, ib txhia neeg tsim pob zeb; thaum lwm tus tsis [19]. Ntawm recurrentpob zebCov qub, tso zis oxalate ntau dua, qhov citrate excretion qis dua Ib txoj kev tshawb fawb kuj qhia tau hais tias oxalate tuaj yeem pab txhawb chloride, sodium, thiab dej reabsorption hauv cov tubules ze ze thiab qhib ntau txoj hauv kev hauv lub raum epithelial hlwb [20].
Txawm hais tias muaj kev txhim kho loj hauv kev kho mob xws li kev siv hluav taws xob ntawm extracorporeal shock yoj lithotripsy (ESWL), tsis muaj cov tshuaj txaus los kho lub raum calculi [21]. ESWL kev kho mob yog txuam nrog mob raum raug mob vim muaj kev raug mob ntawm lub Shock yoj, thiab kab mob tom qab kev kho mob [22]. Nws kuj ua rau tsis tiavpob zebclearance, thiab tsis tuaj yeem zam qhov tshiabpob zebformations [23], orpob zebrov tshwm sim feem ntau mus txog 60 feem pua [24,25]. Tom qab thawj ntu ntawm apob zeb, tus nqi rov qab yog ntau dua 50 feem pua hauv 10-xyoo [26,27]. Yog li ntawd,lub raum pob zebKev rov tshwm sim yog ib qho kev pheej hmoo loj, uas yog suav tias yog ib qho kev sib tw kho mob uas tsis tau ntsib hauv thaj tsam ntawm urology [28].Pob zebKev tsim cov neeg mob yog qhov ua rau nws rov tshwm sim tom qab thawj zaug kev kho mob [29]. Txawm hais tias cov tshuaj muaj pes tsawg leeg ntawm apob zebcuam tshuam rau kev xaiv ntawm kev cuam tshuam [30], qhib kev phais tseem yog lub hauv paus ntawm kev kho mob [31]. Qee cov nroj tsuag tshuaj tau kawm txog tam sim no tiv thaiv urolithiasis yog qhia hauv Table 1
Hauv Ethiopia, cov neeg mob feem ntau vam khom cov nroj tsuag tshuaj raws li kev kho mob rau ntau yam kab mob xws li urolithiasis. Cov nroj tsuag tshuaj muaj nqis, siv tau, siv tau zoo, muaj kev phiv tsawg dua, thiab muaj txiaj ntsig zoo dua rau tib neeg lub cev [41], piv rau cov tshuaj siv niaj hnub no [42].G. fruticosus(L.) raug xaiv los tshuaj xyuas lawv cov teebmeem ntawm kev sim ntxias urolithiasis. Kev soj ntsuam cov ntaub ntawv ntxaws tau ua tiav los xyuas kom meej tias tsis muaj ib qho ntawm cov nroj tsuag tshuaj uas tau xaiv tau kawm txog tam sim no ntawm kev ua haujlwm tiv thaiv urolithiatic. Txog rau hnub tim, tseem tsis muaj pov thawj tshawb fawb tsis txaus qhia txog kev ua haujlwm antihero-lithiatic ntawm cov nroj tsuag tshuaj no. Yog li ntawd, lub hom phiaj ntawm txoj kev tshawb no yog los soj ntsuam cov kev ua tau zoo ntawm cov nroj tsuag tshuaj nyob rau hauv kev sim induced nephrolithiatic txiv neej nas.
Table 1 Cov nroj tsuag tshuaj muaj cov tshuaj tiv thaiv urolithiatic hauv kev tshawb fawb yav dhau los
Cov ntaub ntawv thiab cov txheej txheem
Kev txheeb xyuas thiab sau cov khoom cog tshuajGomphocarpusfruticosus (G. fruticosus) nplooj tau sau thaum lub sij hawm nws flowering los yog fruiting lub sij hawm. CovG.fruticosus(L) Ait.f, lub npe hu ua Tifriena (lub zos lub npe), tau sau ntawm Bole Bulbula nyob ib ncig ntawm "93 Mazoria"Addis Ababa thaum Lub Kaum Hli-ber 2018.G. fruticosusL.Ait.f (Asclepiadaceae) (qhia los ntawm cov neeg qhia tseem ceeb) tau thov rau kev kho mob urolithiasis. Cov qauv cog tau xa mus rau National Herbarium, Department of Plant Biology thiab Biodiversity Management, Addis Ababa University (AAU) rau taxonomic authentication (TA238), thiab tus lej sau tau raug muab. Cov qauv no tau muab tso rau hauv National Herbarium ntawm AAU rau yav tom ntej siv.
Kev npaj ntawm crude extract
Cov ntaub ntawv cog qoob loo raug ntxuav kom huv si nrog cov dej kais dej kom tshem tawm cov kab mob thiab ziab hauv ib lub tsev ntawm qhov kub ntawm 2 mus rau 3 lub lis piam hauv Biomedical Sciences Laboratory, AAU. Cov nroj tsuag qhuav qhuav tau hmoov zoo siv lub tshuab ua noj (mortar thiab pestle, qhov loj li ntawm 9 in.in inch). Cov hmoov tau muab tso rau hauv ib lub sieve ntawm 3 mesh qhov ntau thiab tsawg thiaj li lim cov khoom hnyav.
Cov extracts tau npaj siv cov txheej txheem zoo ib yam li feem ntau siv los ntawm cov kws kho mob lossis cov neeg mob, nrog qee qhov kev hloov kho me me. Cov hmoov nplej tau tsau rau hauv dej distilled, uas tau muab tso rau ntawm lub shaker rau 72h. Cov sib tov tau lim los ntawm paj rwb / gauze, tom qab ntawd los ntawm Whatman lim ntawv xov tooj 1 (pore loj: 1l μm) kom tshem tawm cov khoom cog zoo los yog cov khoom tsis sib haum. Tom qab ntawd, cov semi-solid concentrates poured rau hauv iav petri-phaj thiab tso cai kom qhuav qhuav nyob rau hauv dej da dej hloov mus rau 45 degree, cov qhuav qhuav extracts tau sau thiab khaws cia nyob rau hauv daim ntawv lo sterile fwj npog nrog ib tug nruj stopper thiab khaws cia nyob rau hauv. lub freezer ntawm -20 degree kom txog rau thaum siv hauv kev sim.
Kev npaj ntawm kev ua tiav cov kuab tshuaj extractions
Gomphocarpus fruticosusniam extract (aqueous) tau muab faib nrog cov kuab tshuaj ntawm kev nce polarity los ntawm cov tsis-polar mus rau polar solvents kom ntseeg tau tias ntau qhov polarity ntau ntawm cov tebchaw tuaj yeem muab rho tawm. Kev ua tiav tiav tau pib ua ntu zus nrog roj av ether, tom qab ntawd los ntawm chloroform, ethyl acetate, butanol thiab dej, uas cov kuab tshuaj no tau raug tshem tawm thiab ua kom muaj zog hauv lub tshuab nqus tsev vacuum. Cov khoom siv cog macerated hauv dej distilled tau lyophilized hauv qab lub tshuab ziab khaub ncaws kom tau cov hmoov seem. Cov txiv hmab txiv ntoo qhuav tau capped nruj nreem nrog stoppered thiab muab cia rau hauv iav fwj thiab khaws cia rau hauv lub tub yees ntawm -20 degree C kom txog thaum siv rau kev sim.
cistanche deserticola txiaj ntsig: kho mob raum
Cov tshuaj thiab cov tshuaj reagents / tus qauv tshuaj
Cov tshuaj thiab cov tshuaj reagents los yog cov khoom siv uas siv tau yog cov qhab nia ntsuam xyuas yuav los ntawm ntau qhov chaw. Kev yuav cov roj av ether, chloroform, ethyl acetate, butanol thiab ethanol yog los ntawm Wisteam PLC (Addis Ababa, Ethiopia), potassium dihydrogen phosphate (anhydrous), sodium phosphate (dibasic anhydrous extra pure), Tris-HCl tsis (CaHu), thiab sodium chloride (NaCl) tau yuav los ntawm Micron International Trading House PLC (Addis Ababa Ethiopia). Ib yam li ntawd, isoflurane, thiab formaldehyde tau yuav los ntawm Neway Chemicals PLC (Addis Ababa, Ethiopia). Ethylene glycol (EG) thiab ammonium chloride (NH, Cl) tau yuav los ntawm Pharma PLC (Addis Ababa, Ethiopia). EDTA raj, cov hlab ntsha cais, cov hlab ntsha capillary, thiab cov hniav phais tau yuav los ntawm Micro Pharma PLC (Addis Ababa, Ethiopia). Tsis tas li ntawd, cov khoom siv rau kev kuaj mob siab thiab raum tau yuav los ntawm Roshi PLC (London, England). Cov oxalate (oxalic acid) colorimet-ric assay kits thiab citrate colorimetric/fluorometric assay muas los ntawm BioVision Incorporated kits yog (Milpitas, USA). Potassium citrate hmoov tau los ntawm Lub Tsev Kho Mob Dub Lion Referral (Addis Ababa, Ethiopia).Cystone (polyherbal formulation) tau yuav los ntawm Mumbi, Is Nrias teb. Ascorbic acid, uas tau yuav los ntawm Micron International Trading House PLC (Addis Ababa, Ethiopia).
Solubilization ntawm cov nroj tsuag extract thiab cov qauv tshuaj Cov kev sim extracts / tshuaj ntawm txawv concentrations tau yaj siv cov tsheb tsim nyog, uas yog distilled dej thiab 3 feem pua Tween 80.Txhua cov nroj tsuag rho tawm thiab potassium citrate tau yaj hauv dej distilled ntawm txawv concentration. Cov txheej txheem siv los yaj Cystone zoo ib yam li cov txheej txheem ntawm Pha-tak thiab Hendre [43] thiab Garimella li al. [44] nrog qee qhov kev hloov kho. Cystone ntsiav tshuaj tau hmoov siv ib lub Mortar thiab Pestle (qhov loj me: 0.23m), thiab yaj (tshem tawm) hauv dej distilled (900ul) thiab 100 ul ntawm 3 feem pua Tween 80 (uas yog, 750 mg / ml). Tom qab ntawd, lawv tau khaws cia rau 3h kom yaj, centrifuged ntawm 1000rpm rau 5 min, thiab lim los ntawm 0.22 hli pore loj lim ntawv. 45 ml), huab cua nruj thiab khaws cia rau hauv lub tub yees kom txog thaum kev sim siv. Cov tshuaj ntsuam xyuas thiab cov qauv tshuaj tau npaj txhua hnub thiab tsis ntev ua ntej kev sim tshuaj ntawm koob tshuaj 200 mg / kg ntawm extracts, 750mg / kg ntawm Cystone, thiab 2.5g / kg. Cov khoom siv tshuaj ntsuam xyuas yog 2ml / 100g ntawm lub cev hnyav, dej deionized tau siv los npaj tag nrho cov khoom siv / tshuaj thiab cov no tau npaj txhua hnub ua ntej kev sim tshuaj.
Nyob rau hauv vivo urolithiatic pharmacological kev tshawb nrhiav
Lub hom phiaj tsiaj: noj qab nyob zoo txiv neej albino Wistar nas ntawm tib lub hnub nyoog-pab pawg ntawm 8 mus rau 10 lub lis piam thiab hnyav (220-280g) tau siv, yuav los ntawm Ethiopian Public Health Institute (EPHI), thiab yug ntawm Biomed-ical Sciences Tsiaj Laboratory, AAU. Qhov kev sim tau ua nyob rau ntawm Biomedical Sciences Laboratory ntawm AAU thiab EPHI, raws li kev lees paub thoob ntiaj teb cov txheej txheem rau kev siv tsiaj hauv kev tshawb fawb. Ua ntej pib qhov kev sim, cov nas tau ua kom zoo rau cov qauv kuaj mob (6 nas rau ib lub tawb polypropylene) rau 7 hnub. Lawv tau khaws cia nyob rau hauv qhov chaw tswj ntawm qhov kub thiab txias (27 ± 2 degree), txheeb ze av noo (55 ± 5 feem pua), thiab lub teeb (12h lub teeb / tsaus voj voog). Cov nas no tau noj nrog cov pellets tsis tu ncua (kev noj zaub mov zoo) thiab pub dej haus dawb (ad libitum) rau 28 hnub.
cistanche deserticola txiaj ntsig: kho mob raum
Urolithiasis induction
Lub raum pob zebraug ntxias siv ethylene glycol (EG) nrog rau ammonium chloride (NHCl) uas siv rau hauv cov nas haus dej. Hauv cov qauv hyperoxaluria no, 1 feem pua (w/v) NHACl tau muab nrog 0.75 feem pua (v/v)EG rau thawj 5 hnub txhawm rau txhawm rau txhawm rau lithiasis, tom qab qhov no cov dej tau hloov mus rau {{4 }}.75 feem pua EG ib leeg rau 25 hnub tom ntej [45-47]. Kev nthuav tawm ntawm cov koob tshuaj no tau txaus siab rau hauv kev tshawb fawb tsiaj [48]. EG kev tswj hwm ua rau muaj hyperoxaluria, uas ua rau CaOx deposition hauv ob lub raum [49]. Cov nas sim tau muab raws lipob zebpab pawg kho mob tau txaispob zebinducing kev kho mob rau 28 hnub.
Curative teebmeem ntawm urolithiasis
Hauv kev kho mob (kev sib cais), tag nrho ntawm 54 al-bino Wistar txiv neej nas tau muab faib ua 9 pawg suav nrog 6 tus neeg hauv ib pab pawg nrog qhov sib tw ntawm lub cev hnyav. Hauv kev kho mob kho mob, tus kab mob tau ua ntej los ntawm hnub 1 txog 14. Tom qab ntawd, txhua yam ntawm ex-tracts/drugs tau muab qhov ncauj los ntawm hnub 15 txog 28 concurrently nrog kab mob induction (EG) raws tu qauv los txiav txim curative teebmeem [50, 51] . Thaum kawg ntawm 28th hnub, nas tau txi rau kev tshawb fawb biochemical thiab histopathological. Cov kev sim tsim tau raug xaiv raws li Pawg I (ib txwm tswj), Pab Pawg II (Lithia-tic tswj), Pawg I (K-Cit), Pawg IV (Cystone), Pawg V (G. fruticosusPET extract), Pawg VI (G.fruti-tus nqi Chl. extract), Pawg VI (G. fruticosusEtOAc ex-tract), Pawg VIII (G.fruticosus BuOH extract), thiab Pawg IX(G. fruticosusaq. feem). Lwm cov neeg tshawb xyuas kuj tau siv cov tshuaj potassium citrate (2.5g / kg) los ua kev tswj hwm zoo [52, 53]. Cov koob tshuaj yog 2 ml / 100 g ntawm lub cev hnyav. Pawg tswj hwm tau txais cov dej distilled ib hnub ib zaug thoob plaws qhov kev sim. Thaum kawg ntawm 28th hnub, nas tau txi rau kev tshawb fawb biochemical thiab histopathological.
Kev sau cov zis thiab microscopic tsom xam
Cov zis, ntshav, thiab histological profiles yog cov cim ntawmpob zebtsim thiab rov tshwm sim. Kev nplua nuj thiab morphology ntawm calcium oxalate crystals tsim nyob rau hauv vivopob zebinductions tau tshuaj xyuas siv lub teeb tsom iav (Wagtech Thatcham Berkshire RG194QD, United Kingdom, 40x magnification). Cov nas tsuag tau muab tso rau hauv cais cov kab mob metabolic thiab raug rau 24 teev tso zis thaum hnub 28 rau kev kuaj mob [50]. Hauv kev tshuaj xyuas crystalluria, kwv yees li 3 ml ntawm cov zis tshiab tau muab tso rau hauv lub khob raj thiab centrifuged ntawm 3000rpm rau 10 min kom tshem tawm cov khib nyiab, thiab cov supernatants tau muab pov tseg. Cov tshuaj tso zis no tau siv los txiav txim CaOx crystal tsim. Kwv yees li 10 ul ntawm cov vortexed sediments tau muab tso rau hauv lub iav tsom iav (pob nrog lub npog npog), thiab tshuaj xyuas rau CaOx crystals xav txog lawv cov naj npawb thiab qhov loj me nyob rau hauv lub teeb microscope (40x). Cov muaju uas tsim los ntawm cov tshuaj nyob rau hauv cov zis tau ua tib zoo tshuaj xyuas los ntawm lwm cov khoom tso zis. Cov duab ntawm kev soj ntsuam microscopic tau siv lub koob yees duab digital (Sony Cyber-shot DSC-W180 10.1MP nrog 3x optical zoom, New Jersey, USA) manually mounted rau saum nws.
Biochemical tsom xam cov zis
Thaum kawg ntawm lub sijhawm kho mob, cov tsiaj tau muab tso rau hauv ib lub tawb hauv cov kab mob metabolic, thiab 24 teev cov zis (acidified thiab non-acidified) kuaj. Cov zis tau acidified nrog 1ml ntawm 6N HCL (hydrochloric acid) thiab khaws cia ntawm 4 degree rau 5 hnub. Tom qab ntawd, cov no tau centrifuged ntawm 3000 rpm rau 10 min (REMI, R24), thiab cov supernatant ntawm acidified tso zis tau siv los kwv yees excretions xws li oxalate, calcium, magnesium, thiab phosphate ntsiab lus. Hauv cov zis uas tsis yog-acidified xws li citrate, creatinine, thiab uric acid, thiab tag nrho cov protein ntau tau raug tshuaj xyuas los ntawm kev siv cov khoom siv tshuaj ntsuam xyuas los ntawm Automated Clinical Chemistry analyzer. Ob qho tib si oxalate thiab citrate concentrations tau soj ntsuam los ntawm kev siv ntau lub tshuab spectrophotometer (ELISA nyeem ntawv) raws li phau ntawv qhia nrog cov khoom siv. Rau lub hom phiaj ntawm quantifying, calibration nkhaus tau npaj siv Oxalate thiab Citrate cov khoom siv raws li tus qauv.
Serum collection thiab tsom xam
Tom qab qhov kawg ntawm lub sijhawm sim (hnub 28th), nas raug tshuaj loog siv Isoflurane thiab 3 ml cov ntshav kuaj tau raug sau los ntawm cov hlab ntsha retro-orbital los ntawm capil-lary puncturing. Cov ntshav tau muab cais tawm tom qab centrifuged ntawm 3000rpm, 20 degree degree rau 15 min. Cov tshuaj uas tau sau tseg tau tshawb xyuas txog biochemical tsis xws li creatinine, ntshav urea nitrogen, uric acid, urea, calcium, magnesium, thiab phosphate los ntawm Clinical Chemistry Auto-analyzer (Cobas 6000 analyzer, Germany) nrog rau cov khoom siv kuaj mob. Cov oxalate thiab citrate concentrations ntau qhov kev txiav txim siab siv 1 yog spectrophotometer (ELISA nyeem ntawv) raws li phau ntawv qhia nrog cov khoom siv. Rau lub hom phiaj ntawm quantifying, calibration nkhaus tau npaj siv Oxalate thiab Citrate cov khoom siv raws li tus qauv.
cistanche deserticola txiaj ntsig: tiv thaiv kab mob
Lub raum homogenate tsom xam
Thaum kawg ntawm lub sij hawm sim (hnub 28th), nas tau txi nyob rau hauv Isoflurane tshuaj loog, tom qab ntawd los ntawm ncauj tsev menyuam dislocation. Tom qab ntawd, lub plab tau qhib thiab ob lub raum tau sau los ntawm txhua tus nas. Lub raum cais tau ua tib zoo muab tshem tawm thiab ntxuav (ntxuav / yaug) los ntawm cov ntaub so ntswg extraneous nrog cov dej khov-txias physiological saline tov (0.15 M NaCl). Lub raum sab laug los ntawm txhua tus tsiaj tau khaws cia hauv 10 feem pua tsis muaj formalin thiab siv rau kev tshawb fawb histological. Cov ob lub raum txoj cai raug muab hlais rau hauv ob qho tib si siv cov hniav, thiab ib nrab qhuav ntawm 80 degree hauv qhov cub cua kub. Txoj kev siv tau zoo ib yam li Ashok li al. [35] nrog qee qhov kev hloov kho, uas qhov hnyav ntawm 200 mg (29 feem pua) ntawm tag nrho lub raum qhov hnyav (0.67 g) tau muab tso rau hauv 10 ml ntawm 1N hydrochloric acid (1% HCl), uas tau muab tso rau hauv ib tug boiling dej da dej ntawm 100 degrees C rau 30 feeb. Tom qab ntawd, nws raug txiav npluav rau hauv daim ntawv siv cov hniav, tsoo los ntawm pestle thiab mortar, thiab ntxiv homogenized rau 10 min siv Ultra-Sonicator. Lub homogenate tau centrifuged ntawm 3000 rpm rau 15min, thiab cov supernatant tau sau los ntawm Cryotubes. Thaum kawg, cov supernatants tau siv rau kev kwv yees ntawm calcium, oxalate thiab phosphate cov ntsiab lus nrog cov khoom siv biochemical muaj muag (BioVision PLC, USA), raws li cov chaw tsim khoom raws tu qauv [54,55].
Kev kuaj mob histopathological
Kev kuaj mob histopathological tau ua tiav rau lub raum cov ntaub so ntswg ntawm cov nas sim. Txhua tus nas tau muab txi rau tib neeg txoj kev siv Isoflurane tshuaj loog thaum kawg ntawm hnub 28 (urolithiasis curative studies). Cov ntaub so ntswg tau muab los ntawm ob lub raum thiab tshuaj xyuas rau urolithiatic curative peev xwm ntawm cov nroj tsuag rho tawm. Cov ntaub so ntswg tau kho los ntawm 10 feem pua tsis muaj qhov nruab nrab formalin tov, thiab tom qab ntawd muab tso rau hauv paraffin wax. Cov ntu (5μm tuab) raug txiav siv Rotary Microtome 4060E (Lub Tebchaws Yelemees), mounted rau iav slides, thiab stained nrog Hematoxylin thiab Eosin los kawm txog cov kev hloov pauv hauv histopathological [34]. Tag nrho cov teb ntawm cov ntaub so ntswg morphology raug tshuaj xyuas nyob rau hauv lub teeb microscope (100x magnification) (Wagtech Thatcham, Berkshire, RG19 4}QD, United Kingdom), thiab cov photomi-crographs raug ntes siv lub koob yees duab digital manually mounted nws (Sony Cyber-shot DSC-W180 10.1MP nrog 3x optical Zoom, New Jersey, USA) rau kev siv ntxiv.
Calcium oxalate crystal depositions
Hauv ob lub raum, crystal depositions tau txiav txim siab siv ib nrab-quantitative assays, uas yog ib tug microscopic scoring method [46,56]. Lub microscope filar micrometer (0.1 hli) qhov muag (10x, Dav ntawm 23.3 hli, Olympus Optics OSM 212422, ua hauv Nyij Pooj) tau siv. Ntawd yog, crystal deposits nyob rau hauv lub raum cov ntaub so ntswg raug suav siv lub teeb microscope. Cov qib hnyav ntawm crystal deposits raug muab ua 0=<1 crystal(no="" crystal="" deposition),1="1-10,2=11-30," 3="31-50," 4="51-75" and="" 5="">75 siv lead ua suav, noj tus nqi nruab nrab [46].
Hauv suav CaOx crystal deposits, ib seem sagittal ntawm txhua lub raum kuaj tau muab faib ua plaub thaj tsam sib npaug zos los ntawm ob kab virtual, thiab kev nyeem ntawv ntawm qhov nruab nrab ntawm 4 microscopic teb tau tshaj tawm [57,58].Ib thaj tsam ntawm 100x yog ces randomly. xaiv los ntawm txhua cheeb tsam thiab CaOx deposits raug suav. Daim duab ntawm ib qho ntawm plaub cheeb tsam nyob rau hauv lub teeb tsom iav tau randomly ntes [58] siv lub koob yees duab digital manually mounted rau saum lub Microscope.
Kev txheeb cais
Cov ntaub ntawv tau txheeb xyuas siv Graph Pad Prism version 6 Software (Graph Pad Software, San Diego, CA, USA). Ib txoj kev tsom xam ntawm qhov sib txawv (ANOVA) ua raws li tom qab-hoc Dunnett qhov kev sim sib piv tau ua thaum tsim nyog los sib piv ntawm pab pawg kho thiab tsis kho. Cov ntaub ntawv muaj nuj nqis tau nthuav tawm raws li qhov nruab nrab ± tus qauv sib txawv (SD). Tus nqi ntawm P<0.05 was="" considered="" statistically="">
cistanche deserticola txiaj ntsig: kho mob raum
